PB84-224419
Use of Sewage Sludge on
Agricultural and  Disturbed Lands
Illinois Univ.  at Urbana-Champaign
Prepared  for

Municipal Environmental Research  Lab,
Cincinnati, OH
Jul 84
                     U.S. DEPARTMENT OF COMMERCE
                   National Technical Information Service

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                                                 PB8Q-22.4119
                                        EPA-600/2-84-127
                                        July 1984
USE OF SEWAGE SLUDGE ON AGRICULTURAL AND
             DISTURBED LANDS
                     by

T. D. Hinesly, L. G. Hansen, and D. J. Bray
           University of Illinois
          Urbana, Illinois 61801
              Grant No. R805629
               Project Officer

                G. K. Dotson
        Wasterwater Research Division
 Municipal Environmental Research Laboratory
           Cincinnati, Ohio 45268
 MUNICIPAL ENVIRONMENTAL RESEARCH LABORATORY
    OFFICE OF RESEARCH AND DEVELOPMENT
   U.S. ENVIRONMENTAL PROTECTION AGENCY
         CINCINNATI, OHIO 45268

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                                  TECHNICAL REPORT DATA
                           {Please read luarucnons on the reverse before completing)
I  REPORT NO
   EPA-600/2-84-127
                                                           3. RECIP,
                                                                   i'S I
4 TITLE AND SUBTITLE

  Use of Sewage Sludge on Agricultural and Disturbed
  Lands
             S. REPORT DATE
                 July 1984
             6. PERFORMING ORGANIZATION CODE
7 AUTHOR(S)

  T.  D.  Hinesly, L. G. Hansen,  and D.  J. Bray
                                                          8. PERFORMING ORGANIZATION REPORT NO.
) PERFORMING ORGANIZATION NAME AND AOORESS

   University of Illinois
   Urbana, IL 61801
                                                           1O. PROGRAM ELEMENT NO.
                :AZBIB
  .CAZB
  . CONT!
             11. CONTRACT/GRANT NO.


                Grant No.  R805629
12. SPONSORING AGENCY NAME AND AOORESS
  Municipal Environmental  Kesearch Laboratory  -  Cin., OH
  Office of Research  and  Development
  U.S. Environmental  Protection Agency
  Cincinnati, Ohio  45268
                                                           13. TYPE OF REPORT AND PERIOD COVERED
             14. SPONSORING AGENCY CODE
                EPA/600/14
IS. SUPPLEMENTARY NOTES
  Project Officer:   G.  K.  Dotson (513) 684-7661
16 ABSTRACT
       Results of 8 field  studies of long-term  use  of digested sewage on agricultural
  and disturbed lands are  presented.  The studies included:   (1) response of  corn grown
  on 3 soil types previously amended with annual sludge applications; (2) response
  of corn grown annually on  Blount silt loam treated  annually with sludge;  (3)  contin-
  uous corn on strip mine  spoils treated with sludge; (4) differences in Cd and Zn
  uptake by various corn hybrids; (5) effects of cation exchange capacity on  Cd uptake;
  (6) Cd uptake from Cd-spiked sludge by spinach; (7) response of chickens  to Cd in
  feed; (8) Cd-induced  growth depression and Cd accumulation in chicks as influenced
  by dietary modifications.
       No phytotoxicity developed from trace elements in sludge used annually as
  fertilizer.  Crop uptake of heavy metals from soils containing residual sludge varied
  with species and varieties.  Elevated levels  of dietary Cd did not affect health of
  chickens, egg protection,  nor composition of  the  eggs.
17.
                               KEY WORDS AND DOCUMENT ANALYSIS
                 DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS  C. COSATI Field/Group
13 DISTRIBUTION STATEMEN1

  RELEASE TO PUBLIC
>S iTIus Report)
                           21. NO. OF PAGES

                              250
                                             20 SECURITY CLASS IThnpagel
                                               UNCLASSIFIED
                                                                        22. PRICE
EPA Farm 2220-1 (9-73)

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                                  DISCLAIMER


    Although che information described in this article has been funded wholly
or in part by the United States Environmental Protection Agency through
grant number R805629 to the Metropolitan Sanitary District of Greater Chicago,
it has not been subjected to the Agency's required peer and administrative
review and therefore does not necessarily reflect the views of the Agency and
no official endorsement should be inferred.
                                      ii

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                                    FOREWORD
   The U.S.  Environmental Protection Agency was created because of increasing
public and government concern about the dangers of pollution to the health
and welfare of the American people.  Noxious air, foul water,  and spoiled
land are tragic testimonies to the deterioration of our natural environment.
The complexity of that environment and the interplay of its components
require a concentrated and integrated attack on the problem.

   Research and development is that necessary first step in problem solution,
and it involves defining the problem, measuring its impact, and searching for
solutions.  The Municipal Environmental Research Laboratory develops new and
improved technology and systems to prevent, treat, and manage wastewater and
solid and hazardous waste pollutant discharges from municipal and community
sources, to preserve and treat public drinking water supplies, and to minimize
the adverse economic, social, health, and aesthetic effects of pollution.
This publication is one of the products of that research and is a most vital
communications link between the researcher and the user community.

   This report presents results of a continuing long-term study of methods of
recycling wastewater sludge on land.  It contains information about beneficial
and adverse effects of applying sludge on various kinds of soil at variable
rates and indicates management needed to achieve the greatest benefit with
low degree of risk to the environment or public health.
                                        Francis T. Mayo
                                        Director
                                        Municipal Environmental Research
                                          Laboratory
                                       iii

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                                    ABSTRACT
   Results are presented from eight field studies done during a 15-year
investigation of the long-term use of sewage sludge on agricultural and dis-
turbed lands.  These projects were intended to answer concerns about how
sludge applications to soils relate to phytotoxic accumulations of trace
metals and hazardous metal levels in crops.  Studies were conducted at the
Northeast Agronomy Research Center near Elwood, Illinois.

   Field studies examined the following subjects:  (1) Response of corn on
three soil types previously amended with annual sludge applications, (2)
response of continuously planted corn on Blount silt loam to repeated an-
nual applications of sewage sludge, (3) sludge-amended strip-mine spoils
continuously planted with corn,  (4) differences in Cd and Zn uptake by vari-
ous corn hybrids, (5) effects of cation exchange capacity on Cd uptake, (6)
uptake of metals by spinach from Cd-spiked sludge, (7) response of chickens
to Cd in feed, and (8) Cd-induced growth depression and Cd accumulation in
chicks as influenced by dietary modifications.

   Results showed no evidence that a phytotoxic condition was likely to
develop as a result of the repeated use of the Chicago sludge as a ferti-
lizer.  Losses of sludge-borne heavy metals from soils occurred rather rapidly
by an unknown mechanism.  The availability of residual metal concentrations
in soil from previous sludge applications varied with different crop species
and varieties.  Enhancement of heavy metal concentrations in food and feed
stuffs can be eliminated by plant breeding.  Enhanced levels of dietary Cd
did not affect the health of chickens or increase the levels of the metal
in egg shells, whites, yolks, muscle tissues, or bones.

   This report was submitted in fulfillment of grant No. R805629 by the
University of Illinois and the Metropolitan Sanitary District of Greater
Chicago under the partial sponsorship of the U.S. Environmental Protection
Agency.  This report covers the period October 1977 to December 1980.
                                       iv

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                                    CONTENTS
Foreword	  ill
Abstract 	   iv
Figures 	   vi
Tables 	vii
Acknowledgments 	  xiv

   1.  Introduction 	    1
   2.  Conclusions 	    3
   3.  Recommendations 	    6
   4.  Long-Term Field Studies
         Response of continuous corn grown on three soil types
             previously amended with annual applications of
             digested sewage sludge 	    7
         Response of continuous corn on Blount silt loam to repeated
             annual applications of sewage sludge 	   39
         Response of winter wheat and soybeans on Blount previously
             amended with annual applications of sewage sludge 	   51
         Responses of continuous corn on strip-mine spoil with and
             without annual applications of digested sewage sludge ....   82
   5.  Special Studies
         Changes in strip-mine spoil characteristics and response of
             plants to high-rate sewage sludge applications 	   113
         Differential accumulations of Cd and Zn by corn hybrids
             grown on soil amended with sewage sludge 	   153
         Effect of soil cation exchange capacity on the uptake of Cd
             by corn 	   16 7
         Uptake of metals by spinach grown on soil amended with
             sludge and CdCl. 	   183
         Responses of white leghorn chickens to biologically
             incorporated Cd 	   185
   6.  Abstract of thesis research
         Cadmium-induced growth depression and cadmium accumulation
             in chicks as influenced by dietary modifications  	    230

References 	    231

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                                     FIGURES
Number                                                                 Page

1    pH of saturated extracts from strip-mined spoil without  and
     with various amounts of incorporated sewage sludge 	     128

2    Electrical conductivity of saturated extracts  from strip-mined
     spoil without and with various amounts of incorporated
     sewage sludge 	     129

3    Percent water stable aggregates in strip-mined spoil without  and
     with various amounts of incorporated sewage sludge 	     131

4    Percent moisture retention by strip-mined spoil without  and with
     various amounts of incorporated sewage sludge  	     132

5    Concentrations of Cd in corn tissue grown on soil  mix com-
     binations resulting in three levels of CEC,  containing 10
     mg/kg of Cd derived from CdCl. or sewage sludge 	   173

6    Mean weight (a) and amount of Cd accumulated (b) per plant for
     corn grown for 3 weeks on mixtures of soils combined to  give
     three levels of CEC, containing 10 mg/kg of Cd derived from
     CdCl_ or sewage sludge 	     177

7    Mean weight (a) and amount of Cd accumulated (b) per plant for
     corn grown for 7 weeks on mixtures of soils combined to  give
     three levels of CEC, containing 10 mg/kg of Cd derived from
     CdCl2 or sewage sludge 	     180
                                      vi

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                                     TABLES
Number
      Concentrations of selected chemical elements and soil pH in
      Blount silt loam, Elliott silt loam and Plainfield loamy sand
      lysimeter plots located in a north series 	
  2   Corn grain and stover yields from lysimeter plots of three
      soil types,  with and without sewage sludge irrigations.   All
      three types were represented in a north series 	      26

  3   Concentrations of macro-- and micro-elements in corn leaf,
      grain,  and stover grown on Blount silt loam,  Elliott silt
      loam and Plainfield loamy sand lysimeter plots, with and with-
      out digested sludge irrigations 	      27

  4   Annual  digested sludge loading rates and total accumulations
      on maximum-sludge-treated Blount silt loam plots planted
      to corn	      41

  5   Average contents and total annual amounts of several con-
      stituents of sludge applied on the maximum-treated Blount
      silt loam plots planted to corn.  Appropriately lesser
      amounts of sludge from the same batch were applied on other
      sludge-treated plots on the same day 	      42

  6   Average contents and total amounts of various forms of N and
      ash contents of solids annually applied as constituents  of
      sludge  on Blount silt loam	      43

  7   Concentrations of total macro- and trace-elements, organic-C
      and pH  determined from soil samples collected at depths  of 0
      to 15,  15 to 30, 30 to 46 and 61 to 76 cm in Blount silt
      loam plots 	      44

  8   Plant populations and corn grain and stover yields from
      plots designated NW 800 	      50

  9   Average rainfall at the Northeast Agronomy Research Center
      during  the three growing seasons.  The 1941 to 1970 average
      rainfall for the area is presented in comparison 	      51

 10   Average contents of macroelements and minor elements in
      corn leaf, stover and grain tissue samples from Blount silt
      loam plots 	      52

                                       vii

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Number                                                                 Page

 11    Total contents of selected elements and pH determined in
       0 to 15, 15 to 30, 30 to 46 and 61 to 76 cm soil depth
       samples from Blount silt loam plots planted to soybeans
       and wheat 	.'	     65

 12    Soybean yields and wheat grain and stover yields from
       plots designated NW 500 	     81

 13    Concentrations of selected chemical elements of leaf and
       petiole, bean and stalk samples from soybeans (Beeson cul-
       tivar)  and leaf, grain and stover samples from wheat (Abe
       variety) grown on Blount silt loam, with and without sludge..     83

 14    Concentrations of total annual amounts of several constituents
       contained at relatively high concentrations in sludges applied
       on plots of strip-mine spoil material planted to corn 	     96

 15    Concentrations and total annual amounts of several metals in
       sludges applied on plots of strip-mined spoil materials
       planted to corn 	     96

 16    Total N, organic-C, pH and total concentrations of macro-
       elements and minor elements in 0 to 15,  15 to 30, 30 to 46
       and 61 to 76 cm depths of strip-mined spoil material in plots
       located in Fulton County, Illinois 	     97

 17    Plant populations and grain and stover yields of corn grown
       on strip-mined spoil material with and without digested
       sewage sludge in 1978 	    106

 18    Concentrations of selected chemical elements in corn plant
       tissues from plots of strip-mined spoil material with and
       without sludge	    107

 19    Concentrations of selected chemical elements in sludge and
       the 0 to 15 cm depth of sludge-amended spoil bank material.
       Beginning in October of 1979, separate spoil samples were
       taken from plots planted to grasses (Gr) and corn (Co) 	    116

 20    Concentrations of selected chemical elements and pH of strip-
       mine spoil materials in 0 to 15, 15 to 30, 30 to 46, 46 to 61
       and 61 to 91 cm depths calculated prior to sludge application.
       These data are means of composite samples taken from three
       replicated blocks of four experimental plots each 	    121

 21    Concentrations of selected chemical elements in sludge-
       anended spoil bank materials in 0 to 15, 15 to 30, 30 to 46,
       46 to 61 and 61 to 90 cm depths.  Sludge was applied to plots
       in summer of 1978.  Samples were taken from corn and grass
       plots in 1980 	    122


                                       viii

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Number                                                                 Page

 22    Grain and stover yields for corn, wheat,  and rye and plant
       populations for corn on spoil banks with  and without sludge.
       Corn was planted in dead wheat (W) and.rye (R)  mulch with a
       no-till planter 	     133

 23    Concentrations of selected elements in corn leaf from plots
       with and without sludge.  Corn was planted with a no-till
       planter in dead wheat and rye 	     134

 24    Concentrations of selected elements in corn grain from plots
       with and without sludge.  Corn was planted with a no-till
       planter in dead wheat and rye 	     137

 25    Concentrations of selected elements in rye grain and stover
       from plots with and without sludge 	     139

 26    Concentrations of selected elements in wheat grain and stover
       from plots with and without sludge 	     141

 27    Concentrations of selected elements in leaves of rye and wheat
       from plots with and without sludge 	     144

 28    Concentrations of selected elements in sorghum whole plant from
       plots with and without sludge.  Sorghum was planted in dead
       wheat and rye in July of 19 79 	     146

 29    Concentrations of selected elements in various grasses grown
       in 1980 on plots with and without sludge  	     147

 30    Corn leaf and grain (1979) concentration  ratios for comparing
       the uptake of indigenous elements to those added as constituents
       of sludge	     152

 31    Comparison of Cd concentrations in whole  plants (1st harvest)
       and leaves (2nd harvest) of selected corn inbreds when paired
       with themselves or different inbreds in pots of soil amended
       with sewage sludge 	     157

 32    Concentrations (mg/kg dry weight) of Cd and Zn in leaves of
       corn inbreds and reciprocal crosses grown in pots of silt loam
       amended with sewage sludge.  Inbreds listed first in single-
       crosses were used as females 	     158

 33    Annual amounts of liquid digested sewage  sludge applied on
       maximum-treated Blount plots and accumulative amounts of
       solids, Cd, and Zn constituents of. sludge 	     160

 34    Concentrations of Cd and Zn in Blount silt loam plots, with
       and without annual sewage sludge applications.  Sludge ap-
       plications were initiated in 1969 	     161

                                       ix

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Number                                                                 Page

 35    Grain (adjusted to 15.5% moisture) and stover (dry weight)
       yields produced by two corn hybrids on split-plots of  Blount
       silt loam with and without sewage sludge 	     162

 36    Concentrations (dry weight) of Cd in leaves,  grain and stover
       of two corn hybrids grown on split-plots of Blount silt loam,
       with and without sewage sludge	     164

 37    Concentrations (dry weight) of Zn in leaves,  grain,  and stover
       of two corn hybrids grown on split-plots of Blount silt loam,
       with and without sewage sludge 	     165

 38    Characteristics of experimental soils 	     169

 39    Initial and post-harvest (P-H) characteristics  of soil mix-
       tures amended with either sewage sludge or CdCl-  	     171

 40    Analysis of variance for Cd concentrations in plants (mg/kg)
       harvested 3 weeks after planting 	     174

 41    Analysis of variance for mean total amounts (mg)  of Cd ac-
       cumulated per plant during the first 3 weeks  	     178

 42    Analysis of variance for total dry weight (g)  in  plants
       after 7 weeks of growth 	     179

 43    Analysis of variance for mean total amounts (mg)  of Cd ac-
       cumulated per plant during 7 weeks of growth  	     181

 44    Total solids and cadmium content of the 10 sludges used in
       the greenhouse spinach experiment 	     184

 45    Concentrations of selected chemical elements  (dry weight) in
       spinach grown on Blount silt loam amended with  sewage  sludge,
       sludge plus CdCl. and CdCl2 at rates to provide equivalent
       amounts of total soil-Cd.   Sludge and sludge  plus CdCl. con-
       tained 40 mg/kg (dry weight) of Cd	     186

 46    Concentrations of selected chemical elements  (dry weight) in
       spinach grown on Blount silt loam amended with  sewage  sludge,
       sludge plus CdCl- and CdCl. at rates to provide equivalent
       amounts of total soil-Cd.   Sludge and sludge  plus CdCl. con-
       tained 140 mg/kg (dry weight) of Cd	     187

 47    Concentrations of selected chemical elements  (dry weight) in
       spinach grown on Blount silt loam amended with  sewage  sludge,
       sludge plus CdCl2 and CdCl2 at rates to provide equivalent
       amounts of total soil-Cd.   Sludge and sludge  plus CdCl- con-
       tained 400 mg/kg (dry weight) of Cd 	     1£8

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Number                                                                 Page

 48    Concentrations of selected chemical elements (dry weight)  in
       spinach grown on Blount silt loam amended with sewage sludge,
       sludge plus CdCl. and CdCl. at rates to provide equivalent
       amounts of total soil-Cd.  Sludge and' sludge plus CdCl. con-
       tained 600 mg/kg (dry weight) of Cd 	     189

 49    Concentrations of selected chemical elements (dry weight)  in
       spinach grown on Blount silt loam amended with sewage sludge,
       sludge plus CdCl- and CdCl- at rates to provide equivalent
       amounts of total soil-Cd.  Sludge and sludge plus CdCl. con-
       tained 1,000 mg/kg (dry weight)  of Cd.   Sludge I was composed
       of sludge with 40 mg Cd/kg plus  CdCl.;  Sludge II was composed
       of sludge with 140 mg Cd/kg plus CdCI2  	     190

 50    Experimental diet 	     192

 51    Mean concentrations of selected elements in feed samples of
       low-,  medium-, and high-Cd diets formulated from corn and
       soybeans grown on sludge-amended soil 	     193

 52    Grams  of feed consumed per bird  per day (g/b/d) during 2- or
       4-week interval periods of the 80-weeks study 	     196

 53    Cumulative gains in body weight  of survivors for indicated
       periods 	     19 7

 54    Percent of surviving hens laying an egg per day (HD%) from
       20-80  weeks by 4-week intervals  	     198

 55    Disposition of birds, 0-80 weeks of age 	     199

 56    Mean Cd concentrations in egg constituents sampled at various
       intervals during a 54-week period commencing when egg laying
       began	     200

 57    Concentrations of selected elements in  various tissues of hens
       fed low-, medium- and high-cadmium diets formulated from corn
       and soybeans grown on sludge-amended soil.  Samples taken 8
       weeks  after hatching	     201

 58    Concentrations of selected elements in  various tissues of hens
       fed low-, medium- and high-cadmium diets formulated from corn
       and soybeans grown on sludge-amended soil.  Samples taken 8
       weeks  after hatching 	     202

 59    Concentrations of selected elements in  various tissues of hens
       fed low-, medium- and high-cadmium diets formulated from corn
       and soybeans grown on sludge-amended soil.  Samples taken 20
       weeks  after hatching 	     203
                                       xi

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Number
 60    Concentrations of selected elements in various tissues of hens
       fed low-, medium- and high-cadmium diets formulated from corn
       and soybeans grown on sludge-amended soil.  Samples taken 20
       weeks after hatching 	    204

 61    Concentrations of selected elements in various tissues of hens
       fed low-, medium- and high-cadmium diets formulated from corn
       and soybeans grown on sludge-amended soil.  Samples taken 50
       weeks after hatching 	    205

 62    Concentrations of selected elements in various tissues of hens
       fed low-, medium- and high-cadmium diets formulated from corn
       and soybeans grown on sludge-amended soil.  Samples taken 50
       weeks after hatching	    209

 63    Concentrations of selected elements in various tissues of hens
       fed low-, medium- and high-cadmium diets formulated from corn
       and soybeans grown on sludge-amended soil.  After 50 weeks,  sub .
       groups from those hens being fed the low- and high-cadmium
       diets were switched to high- and low-cadmium diets, respec-
       tively.  Samples taken 72 weeks after hatching 	    211

 64    Concentrations of selected elements in various tissues of hens
       fed low-, medium- and high-cadmium diets formulated from corn
       and soybeans grown on sludge-amended soil.  After 50 weeks,
       sub groups from those hens being fed the low- and high-cadmium
       diets were switched to high— and low-cadmium diets, respec-
       tively.  Samples taken 80 weeks after hatching 	    213

 65    Hematological parameters in white leghorn hens having re-
       ceived sludge-fertilized corn-soybean diets from 1 week
       of age 	    217

 66    Red blood cell characteristics in white leghorn hens having
       received sludge-fertilized corn-soybean diets from 1 week
       of age 	    219

 67    Leukocyte populations in white leghorn hens having received
       sludge-fertilized corn-soybean diets from 1 week of age 	    221

 68    Serum chemistries from white leghorn hens having received
       sludge-fertilized corn-soybean diets from 1 week of age 	    222

 69    Relative liver, gizzard, and kidney weights in white leghorn
       hens having received sludge-fertilized corn-soybean diets
       from 1 week of age 	    224

 70    Relative organ weights in white leghorn hens having received
       sludge-fertilized corn-soybean diets from 1 week of age 	    226
                                       xii

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Number                                                                 Page

 71    Fat content of livers from white leghorn hens having received
       sludge-fertilized corn-soybean diets from 1 week of age 	    227

 72    Hepatic microsomal parameters in white leghorn hens having
       received different levels of dietary Cd from 1 week of age ..    228

 73    Hepatic microsomal 0-dealkylation of £-nitrophenetole in
       white leghorn hens having received dietary Cd from 1 week
       of age 	    229
                                        xiii

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                                 ACKNOWLEDGMENTS


Contributions to the project were made by the following principal
investigators:

   Dr. D. E. Alexander, Department of Agronomy
   Dr. J. L. Dorner, College of Veterinary Medicine
   Dr. J. Simon, College of Veterinary Medicine
Special thanks are also due to the following former and present staff
members for their contributions to this research:
   College of Agriculture:

        Mr. E. L. Ziegler
        Dr. K. E. Redborg
        Mr. G. T. Kesner
        Dr. R. D. Rowland
        Mr. D. B. Atkins
        Mr. J. Cali
        Ms. N. A. Frohan
Mr. D. E. Harshbarger
Mr. R. J. Keigher
Ms. A. M. Moore
Mr. D. L. Mulvaney
Ms. J. A. Plesha
Mr. D. S. Thiel
   College of Veterinary Medicine:

        Ms. S. M. Sunlof
        Dr. R. J. Gardner
        Mr. R. S. Vogel
        Dr. S. F. Sunlof
                                       xiv

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                                    SECTION I
                                  INTRODUCTION

   Digested sewage sludge is an effective source of nitrogen and phosphorus
for Che fertilization of field crops.  When applied at rates sufficient to
provide recommended rates of supplementary nitrogen for nonleguminous crop
plants, sewage sludge also supplies relatively high amounts of organic
matter.  Thus long-term usage of sludge changes the physical properties of
some soil types to such an extent that crop yields frequently exceed those
obtainable with commercial fertilizers.  Agricultural use of sewage sludges
eliminates the high energy costs, potential air pollution, and ash disposal
problems encountered by incineration of sludge.  Sewage sludges contain
trace elements at concentrations that, on a dry weight basis, often greatly
exceed normal concentrations in productive soils.  Many environmentalists
are therefore concerned that heavy metals and metalloids may eventually
accumulate and reach phytotoxic levels.  Some are concerned that concentra-
tions of certain trace elements may be enhanced in food and feed stuffs
to such an extent that they may present a health hazard to man and animals.

   Our research was directed toward providing data needed to evaluate threats
to soil productivity, the environment in general and human health.  The
following is a brief statement of specific objectives which are expressed
in more detail in appropriate sections of the report.

Objective 1.

   Two field studies continued earlier investigations designed to monitor
long-term accumulations of sludge-borne trace elements in soils and strip
mine spoil material and determine their effect on corn yields and uptake by
plants.  Previous results were presented and discussed in two progress
reports submitted to the Metropolitan Sanitary District of Greater Chicago
and the U.S. Environmental Protection Agency (Hinesly et al., 1974; Hinesly
and Hansen, 1981).

Objective 2.

   Two other field studies were done to determine the residual effects of
previous sludge applications on yields of corn, wheat, and soybeans and their
trace element contents.

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Objective 3.

   One new field study was established to determine the effects of sewage
sludge on plant growth, chemical composition of plant tissues, and changes
in the chemical and physical properties of the spoil material when one-time
applications of dewatered, digested sewage sludge were applied at rates far
exceeding those required for maximum crop yields.


Objective 4.

   Another field study was conducted to demonstrate that the uptake and
accumulation of Cd and Zn in above-ground plant parts are consistently deter-
mined much more by the genetic constitution of plants than by the total
concentrations of the metals in soils.
Objective 5.

   Three greenhouse studies provided information about the effects of soil
cation exchange capacity and Cd source on the uptake of Cd by corn and
spinach.


Objective 6.

   The report concludes with a discussion of the results from a long-term
poultry feeding study.  Three levels of Cd,  biologically incorporated in
soybean and corn diets, were fed to chicks and throughout their productive
life as laying hens.  Concentrations of Cd and other elements in various
tissues were periodically determined from sacrificed birds, along with other
measurements and observations for health effects.

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                                    SECTION 2

                                   CONCLUSIONS
   Where digested sludge was annually applied by furrow irrigation for 13
years on Blount silt loam, the growth and yields of two different corn
hybrids indicated no imminent phytotoxic conditions.  This observation held
true even on plots receiving 10 times the sludge application rate that would
supply recommended rates of supplementary nitrogen for corn.  Corn yields
were increased by sludge applications in 2 of the last 3 years of the study.
During the last 3 years of the study, further sludge applications failed
to increase soil contents of organic C, N, K, Na, Ca, Mg, Fe, and Mn, but
they did increase Cd, Cu, Cr, Ni, P, Pb, and Zn at the 0- to 30-cm depth.
Except for Cd and Zn, there was no indication that sludge constituents had
leached below the 30-cm depth.  But Cd and Zn concentrations were not in-
creased in soil depths below 45 cm.  Though nothing indicates that signifi-
cant amounts of heavy and transition metals were lost from the soil profile
by leaching, about 40% of the total amounts added as constituents of sludge
either disappeared from the soil or were not detected by the core sampling
method used in this study.  Only Cd, Cu, Ni, and Zn were consistently higher
in above-ground parts of plants grown on sludge-treated plots as compared
with those on control plots.  But these elements were not increased in
plant tissues by accumulative applications of sludge-borne Cd, Cu, Ni, and
Zn.  For a particular loading rate and corn hybrid, contents of these heavy
metals remained fairly constant after about 3 years of sludge application.
There was no indication that a phytotoxic condition is likely to develop as
a result of the repeated use of the Chicago sludge as a fertilizer.

   Where the protocol for a sludge study on calcareous strip-mine spoil was
the same as that on acid Blount silt loam, as discussed above, the results
were similar.  The one exception was that Cd and Zn concentrations in corn
plant tissues increased as a result of accumulative sludge-borne applica-
tions of these metals.  The high pH of calcareous spoil failed to control
the uptake of Cd and Zn by corn to levels lower than those observed on acid
Blount silt loam.

   Where one-time applications of dewatered, digested sludge were made on
level strip-mine spoil at rates of 0, 224, 448, and 896 mt/ha, plant growth
and yields were increased by the two intermediate loading rates as compared
with those on highly fertilized control plots.  Organic C, N, P, and heavy
metals were increased in the 0- to 18-cm depth of spoil in proportion to
sludge loading rates.  Spoil pH decreased, electrical conductivity of water
extracts increased, the percent of water-stable aggregates increased, and the
water-holding capacity of spoil increased in proportion to higher sludge
loading rates.  Two years after sludge applications were made, organic-C

-------
and N contents remained close to initial post application levels in spoil;
but heavy metal contents had decreased in the spoil surface.   Concentrations
of Cd, Cu, and Fb were increased in spoil below the depth of  mixing,  but
not below the 60-cm depth.  Concentrations of Cd, Cu,  Fb, Ni, and Zn in
spoil below the depth of sludge mixing were too low to account for losses
observed in the surface layer.  Concentrations of Mg,  P, Mn,  Cd, Ni,  and Zn
were increased in corn, wheat, and rye grain by sludge applications during
the first year.  During the second year, Ni was not increased in grain by
sludge applications, and Cd contents were considerably less.   Doubling the
448-mt/ha sludge application did not increase grain Cd concentrations, but
it did increase Zn concentrations.  Above the intermediate sludge loading
rate, it appeared that the availability of Cd for plant uptake was limited
by other constituents contained in the sludge.  Concentration ratios did
not indicate that metal concentrations were increased  as a result of low
yields on maximum-sludge-treated plots, except for Ni.  Results from this
study suggest that in view of the low Cd content relative to  concentrations
of essential elements for animals (in particular Cu and Zn),  a one-time
sludge application of about 200 mt/ha may produce better quality grain and
forage than could be obtained with an equivalent amount of sludge applied
over several years at rates to supply recommended amounts of  N.  But special
precaution will be required to protect ground water against contamination
with NO.-N where one-time, high-rate sludge applications are  made.

   On Blount silt loam plots irrigated annually for 5  years with digested
sludge, corn yields began to decline 3 years after the last application.
But applications of N fertilizer at recommended rates  across  all plots
restored soil productivity to the original high level  obtained with sludge
applications.  A year after sludge applications were terminated, organic
C, total N, P, and heavy metal concentrations in Blount tended to remain
rather constant.  But the uptake of heavy metals by corn decreased each
year and 4 years after sludge applications were terminated Cd and Zn con-
centrations in grain were no different from those in grain from control plots,
Data from this study show that regardless of the nature of the mechanism,
the losses of sludge-borne heavy metals from soils occur rather rapidly,
and the amounts remaining for uptake by corn plants decrease  with time and
are not affected by applications of inorganic sources  of N.

   In other studies of the effects of residual sludge  in soils, concentra-
tions of Cd, Cu, Ni, and Zn were increased in soybean  tissues by previous
sludge applications.  By order of listing, Ni, Cd, and Zn concentrations
decreased most rapidly in soybean tissues after annual sludge applications
were terminated.  But 6 years after sludge applications were  terminated,
Cd and Zn concentrations in beans from plots previously treated with sludge
were higher than those in beans from control plots.  In a similar residual
study using wheat, enhanced NI and Cu concentrations in grain from plots
previously treated with sludge returned to background  levels  in 3 years;
but Cd and Zn uptake had not decreased 8 years after sludge applications
had been suspended.  Thus the decrease in availability of residual metal
concentrations in soil from previous sludge applications varies with dif-
ferences in crop species and probably with varieties within a particular
species.

-------
   Results from a field study showed that two commercially available hybrids
selected for their different capacities to take up Cd were indeed different
when grown on split plots that had been irrigated annually with sludge during
the 9 years before the 3-year study.  Except in the first year of the study,
grain Cd concentrations were less in the low-Cd hybrid grown on maximum-
sludge-treated plots than they were in the high-Cd-accumulator grown on
control plots.  Cadmium concentrations in grain from the low-Cd-accumulator
never exceeded the upper range of normal background levels in grain, even
though annual sludge applications were continued throughout the study period.
During the last year of the study, the Cd concentration in grain from the
low-Cd-accumulator grown on maximum-sludge-treated plots was not signifi-
cantly different from concentrations in grain from control plots.  The uptake
of other chemical elements by the two hybrids was independent of their in-
herited capacity to take up or exclude Cd.  Thus enhancement of heavy metal
concentrations in food and feed stuffs can be controlled by plant breeding.

   The results from greenhouse studies showed that differences in soil
cation exchange capacity (CEC) did not affect the uptake of sludge-borne
Cd by corn, but CEC did substantially influence the uptake of Cd from soils
amended with CdCl-.  Where soil CEC was due mainly to organic matter, less
Cd was taken up by corn on CdCl^-amended soil than on similarly amended
soils where the CEC was due to inorganic materials.  The uptake of Cd by
spinach from soils amended with CdCl»-spiked sludges was greater than when
equivalent amounts of Cd were supplied as a natural constituent of digested
sludge.  Thus data from studies involving the use of soluble Cd salts are
of little value in predicting Cd uptake by plants grown on sludge-amended
soils.

   Corn hybrids and soybean cultivars were selected for different capacities
to take up Cd and were grown in a sludge-amended field to obtain distinctly
different levels of Cd concentrations in corn grain and beans.  The corn
grain and processed beans were formulated into starter, developer,  and layer
rations to provide three different levels of biologically incorporated Cd.
In birds sacrificed at 8, 20, 50, 72, and 80 weeks, Cd concentrations in
crop, preventriculus, muscular gizzard, gizzard lining, duodenum, liver,
kidney, pancreas, and spleen tissues paralleled levels of the metal in diets.
But Cd concentrations in leg muscle, breast muscle, and femur bone were un-
related to metal levels in the diets.  For the tissues that accumulated Cd
(except the kidneys), concentrations appeared to reach an equilibrium with
Cd levels in the diet at about 50 weeks of age.  In kidneys, Cd levels con-
tinued to increase regardless of dietary levels.  Also, when birds were
switched from high- to low-Cd diets, Cd concentrations decreased rather
rapidly in all accumulator tissues but the kidneys.  At the highest level
of Cd that could be biologically incorporated in corn grain and soybeans
produced on sludge-amended fields, nothing (body weight changes, egg pro-
duction, or various clinical parameters) indicated that the enhanced levels
of dietary Cd affected the health of the chickens.

   Since no evidence showed that the highest possible level of biologically
incorporated Cd increased levels of the metal in egg shells, whites, or yolks,
or in muscle tissues and bones, the probability of increasing Cd in human
foods to harmful levels is nominal.

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                                    SECTION 3
                                 RECOMMENDATIONS

   The following recommendations are based on Che findings presented in
this report:

   1.  Sewage sludges containing excessive concentrations of heavy metals and
metalloids should be spread only on lands owned and operated by the waste-
water treatment plant authority to ensure that site operation includes
application methods, soil management practices,  selection of appropriate
crops, and crop disposal procedures that minimize potential contamination
of water supplies and recycling of toxic concentrations of trace elements
in food chains.

   2.  To protect ground water supplies from the leaching of NO--N, sludge
loading rates should not exceed amount needed to meet nitrogen fertilizer
recommendations.

   3.  Where excessive leaching of NO--N is prevented by the presence of
slowly permeable geological material, one-time,  high-rate sludge applica-
tions provide several advantages over annual low-rate applications when the
object is to restore severely disturbed lands to a highly productive state.
One-time, high-rate applications provide the opportunity to use various
tillage, crop, and water management techniques that minimize water and wind
erosion more than is possible when sludge is applied annually.  Optimum
sludge-loading rates for one-time applications in humid regions is about
200 mt/ha (dry weight).

   4.  Since most metals and metalloids contained in stabilized digested
sewage sludges will not cause phytotoxic conditions in soils, the main con-
cern is to protect food supplies against excessive concentrations of chemical
elements accumulated by plants.  Particular attention should be given to
minimizing accumulations of Cd and Ni in plants.  More attention should be
given to selecting or producing crop varieties or cultivars by breeding
that can severely limit metal uptake in plant parts used for food while
maintaining adequate concentrations of other essential trace elements.

   5.  When plant materials containing enhanced Cd concentrations are fed
to animals, only the muscle tissues should be consumed by humans.

   6.  When further research on sludge use is funded, priority should be
given to determining how sludge-borne heavy metals are lost from soils.

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                                    SECTION 4
                             LONG-TERM FIELD STUDIES

RESPONSES OF CONTINUOUS COBN GROWN ON THREE SOIL TYPES PREVIOUSLY AMENDED
WITH ANNUAL APPLICATIONS OF DIGESTED SEWAGE SLUDGE
Introduction

   From a review of the most current data, members of a task group (CAST
1980) concluded that "the length of time sludge-derived Cd and Zn remain
available to crops after sludge applications have ceased is indeterminate".
They thought it likely that concentrations of these metals would increase
in plant tissues if the soil pH was allowed to decrease.

   The main objective of this study was to obtain information about the
uptake of transition and heavy metals by one corn hybrid after suspending
further annual applications of sewage sludge.

Methods and Materials

   Forty-four lysimeter plots each 3.05 by 15.25 m in size were established
in 1969 to measure the effects of various application rates of digested,
municipal sewage sludge on soils, crops, surface water, and ground water.
Three soil types were represented in the lysimeter plots.  PIainfield sand,
Elliott silt loam, and Blount silt loam were treated with three rates of
sludge.  About 25.4 mm of liquid sludge was applied on maximum-treated
plots throughout the growing season as often as permitted by weather con-
ditions.  About 12.7 and 6.4 mm of sludge was applied on 1/2- and 1/4-
maximum-treated plots, respectively, on the same day that maximum appli-
cations were made.  Also on the same day, control plots were irrigated with
25.4 mm of well water.  The lysimeters were divided into north and south
groups of 22 plots each.  The north lysimeters were used in this study.
Prior to beginning this study, 232.5 mt/ha of solids, 8,044 kg P/ha, 58.3
kg Cd/ha, and 1290 kg Zn/ha had been applied on maximum-treated plots as
constituents of sludge.  After 5 years sludge applications were terminated
on all plots except for maximum-treated Elliott silt loam and Plainfield
sand.  These plots received an extra 3-years of sludge applications and
through 1976 sludge constituents amounted to 423 mt/ha a/solids, 14,875
kg P/ha, 112.7 kg Cd/ha, and 2,169 kg Zn/ha.

   Details of this study were described in previous publications (Hinesly
et al, 1979; Hinesly and Hansen, 1981).  The same corn hybrid (Pioneer
3517) was planted during years when soils were irrigated with digested

-------
sewage sludge and after applications were suspended.   During the last 2
years, 179 kg/ha of N was broadcast on the entire plot area each spring
before plots were tilled.

Results and Discussion

   The persistence of changes in several soil properties caused by sludge
applications can be seen in Table 1.  Organic-C and N contents in the 0 to
15 cm depth of Blount continued to be significantly higher in sludge-treated
plots and had changed very little since sludge applications were suspended.
After sludge applications were terminated on maximum-treated Elliott and
Flainfield, organic-C and N contents remained fairly stable.  These data
indicate that after the initial flush of mineralization that occurred during
the first year after sludge organic matter was incorporated into soils, the
remaining material was highly resistant to further degradation.  The rate of
degradation was affected very little by differences in soil type.

   Concentrations of K, Na, Ca, Mg, Fe, and Mn in soils were unaffected by
sludge applications (Table 1).  Concentrations of P,  Cd, Cu, Cr, Ni, Pb, and
Zn were significantly increased in the 0 to 15 cm depth of Blount by sludge
applications.  Where sludge was applied for an additional 3 years on maximum-
treated Elliott and Plainfield, concentrations of Cu, Cr, Ni, and Pb were
increased throughout the 0 to 30 cm soil depths.  Zinc concentrations in
these two soils were increased to depths of 46 cm.  Cadmium concentrations
were also increased to depths of 46 cm in Plainfield, and concentrations of
this metal increased in the 61 to 76 cm depth of maximum-treated Elliott
plots.  Thus, the chemical composition of Blount was  changed only in the
plow layer depth by 5 years of repeated sludge applications, but an addi-
tional 3 years of applications on maximum-treated Elliott and Plainfield
resulted in migration of these chemical elements to deeper soil depths.
Although a great deal of year to year variability in concentration of tran-
sition and heavy metals in soils existed,  probably as a result of the in-
adequate number of soil cores collected each year, there is no evidence of
an extensive loss of any chemical element from the soil surface after sludge
applications were terminated.  During the last 3 years of the study, it was
evident that maximum sludge applications had decreased soil pH in all soil
types to a depth of 30 cm.

   Residual effects of sludge applications on Blount  produced an increase
in corn yields during 1976, 1977, and 1978.  But after N fertilizer was
applied in 1979, yields were not significantly different, regardless of
treatment (Table 2).  During the three years after sludge applications were
discontinued, corn yields tended to decline each year until N fertilizer
was applied across all plots.  After N was applied, yields for the last 2
years on Blount were as high as average yields during the 10-year corn study.
Corn was first planted on the north series of lysimeter plots in 1971.
Stover yields on Blount were similar to grain yields  and there were no indi-
cations that residual effects from sludge adversely affected the soil pro-
ductivity.  In the last year of the study, grain yields from Elliott silt
loam were unusually low where maximum amounts of sludge were formerly ap-
plied, but Stover yields from the plot were among the higher ones recorded
during the study.  Grain and Stover yields from Plainfield loamy sand were


                                        8

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    TABLE 1.  CONCENTRATIONS OF SELECTED CHEMICAL ELEMENTS AND SOIL pH IN BLOUNT SILT LOAM, ELLIOTT SILT
              LOAM AND PLAINFIELD LOAMY SAND LYSIMETER PLOTS LOCATED IN A NORTH SERIES.a/
vo














Soil Type
Blount silt
Ana- Depth Year
lyte cm
0-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979
Ck

0.88
0.73
1.00
0.80
0.76
0.85
0.59
0.49
0.33
0.46
1/4
Max

1.23
1.09
1.32
0.60
0.62
0.72
0.48
0.44
0.41
0.47
1/2
Max

1.31
1.54
1.62
0.63
0.62
0.55
0.50
0.42
0.33
0.38
loam
Max

1.60
1.54
1.84
0.77
0.61
0.70
0.47
0.49
0.41
0.41
Elliott silt loam
LSD

n.s.
0.39**
0.47**
n.s.
n.s.
n.s.
n.s.
n.s.
n. s.
n.s.
Ck

Z.
1.54
1.51
1.62
1.25
1.04
1.38
0.44
0.34
0.30
0.34
1/4
Max

1.55
1.68
1.89
1.60
1.63
1.63
0.94
0.66
0.38
0.60
1/2
Max

1.72
1.19
1.87
1.43
1.40
1.37
0.95
0.48
0.44
0.41
Max

3.77
2.60
3.20
1.83
1.36
1.29
0.90
0.59
0.45
0.38
Plainfield loamy sand
Ck

0.42
0.49
0.64
0.22
0.30
0.19
0.06
0.04
0.04
0.08
1/4
Max

0.34
0.47
0.83
0.29
0.17
0.23
0.05
0.12
0.05
0.11
1/2
Max

0.89
0.81
0.52
6.59
0.44
0.20
0.10
0.09
0.01
0.04
Max

3.35
2.10
3.03
0.40
0.46
0.37
0.09
0.01
O.'ll
0.13
                                                    (continued)

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TABLE 1.  (continued)
                                                        Soil Type
Blount silt loam
Ana-
lyte
Depth Year
cm
Ck
1/4
Max
1/2 Max LSD
Max
Elliott silt loam
Ck 1 Ik
Max
1/2 Max
Max
Plain field loamy sand
Ck I/A
Max
1/2
Max
Max
       0-15   1978  0.100 0.119 0.152 0.179 0.018**
              1979  0.1030.1130.1330.1400.023*
              1980  0.101 0.119 0.141 0.156 0.029*
0.147 0.150 0.172 0.350
0.140 0.120 0.170 0.250
0.129 0.164 0.164 0.291
0.041 0.055 0.057 0.342
0.055 0.080 0.090 0.210
0.057 0.069 0.076 0.234
       15-30  1978  0.082 0.077 0.078 0.086  n.s.
              1979  0.087 0.070 0.070 0.070  n.s.
              1980  0.091 0.094 0.071 0.080  n.s.
0.121 0.153 0.132 0.164
0.100 0.150 0.130 0.130
0.073 0.146 0.123 0.127
0.028 0.034 0.054 0.045
0.030 0.020 0.050 0.050
0.016 0.049 0.022 0.042
       30-46  1978  0.079 0.069 0.062 0.061  n.s.
              1979  0.067 0.070 0.050 0.063  n.s.
0.053 0.101 0.097 0.089
0.065 0.080 0.080 0.070
0.014 0.010 0.015 0.021
0.010 0.010 0.020 0.010
       61-76  1978  0.057 0.070 0.064 0.072  n.s.
              1979  0.053 0.060 0.050 0.053  n.s.
0.050 0.068 0.061 0.072
0.045 0.060 0.050 0.050
0.022 0.010 0.006 0.013
0.010 0.010 0.010 0.010
                                                (continued)

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TABLE 1.  (continued)
                                                        Soil Type
Blount silt loam
Ana-
lyte
Depth Year
cm
Ck 1/4
Max
1/2 Max LSD
Max
Elliott silt loam
Ck 1/4
Max
1/2 Max
Max
Plainfleld loamy sand
Ck
1/4
Max
1/2
Max
Max
       0-15   1978  0.078 0.111 0.152 0.217 0.023**
              1979  0.072 0.101 0.123 0.152 0.044**
              1980  0.070 0.113 0.152 0.193 0.057**
0.073 0.100 0.155 0.417
0.067 0.098 0.133 0.258
0.080 0.110 0.114 0.327
0.044 0.077 0.112 0.496
0.056 0.105 0.133 0.308
0.059 0.084 0.086 0.416
       15-30  1978  0.041 0.064 0.039 0.050  n.s.
              1979  0.044 0.066 0.043 0.044  n.s.
              1980  0.049 0.070 0.040 0.058  n.s.
0.044 0.052 0.051 0.101
0.046 0.063 0.054 0.092
0.050 0.052 0.054 0.059
0.039 0.050 0.100 0.082
0.041 0.041 0.081 0.093
0.038 0.041 0.049 0.077
       30-46  1978  0.037 0.049 0.037 0.041  n.s.
              1979  0.031 0.036 0.029 0.032  n.s.
0.041 0.057 0.043 0.068
0.034 0.034 0.025 0.036
0.027 0.022 0.032 0.024
0.024 0.021 0.032 0.028
       61-76  1978  0.041 0.043 0.040 0.041  n.s.
              1979  0.033 0.031 0.037 0.035  n.s.
0.043 0.044 0.042 0.061
0.031 0.036 0.031 0.033
0.020 0.024 0.019 0.039
0.020 0.016 0.015 0.020
                                           (continued)

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    TABLE  1.   (continued)
N)
Soil Type
Ana- Depth Year
lyte cm
K 0-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979

Ck

2.03
2.07
2.01
2.06
2.08
1.98
1.89
2.04
1.97
2.06
Blount
1/4
Max

2.17
2.21
2.08
2.13
2.19
2.13
2.26
2.56
2.34
2.74
silt
1/2
Max

2.12
2.08
2.13
2.09
2.06
2.03
2.05
2.27
2.46
2.39
loam
Max

2.09
2.12
2.00
1.99
1.73
1.94
1.88
1.95
2.43
2.48
Elliott silt loam
LSD

n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
Ck

Z.
1.99
2.04
1.97
1.99
1.97
1.94
1.99
2.39
2.55
2.57
1/4
Max

2.06
2.04
1.93
1.95
2.02
1.91
1.99
2.12
2.44
2.37
1/2
Max

2.08
2.00
1.60
2.01
2.05
1.99
2.11
2.11
2.42
2.48
Max

1.94
1.98
2.06
2.00
2.03
1.97
1.36
2.08
2.01
2.04
Plainfield
Ck

1.50
1.64
1.74
1.55
1.47
1.42
1.33
1.31
1.13
1.33
1/4
Max

1.60
1.69
1.71
1.59
1.29
1.46
1.28
1.27
1.63
1.22
loamy
1/2
Max

1.45
1.42
1.27
1.49
1.30
1.45
1.68
1.31
1.24
1.08
sand
Max

1.63
1.56
1.56
1.46
1.20
1.46
1.11
1.47
1.44
1.26
                                                 (continued)

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TABLE 1.  (continued)
                                                       Soil Type
Blount silt loam
Ana-
lyte
Depth Year
cm
Ck 1/4
Max
1/2 Max LSD
Max
Elliott silt loam
Ck 1/4
Max
1/2 Max
Max
Plainfleld loamy sand
Ck 1/4
Max
1/2
Max
Max
 Na    0-15    1978  0.668 0.671 0.704 0.690  n.s.
              1979  0.749 0.775 0.809 0.784  n.s.
              1980  0.706 0.664 0.758 0.689  n.s.
0.748 0.735 0.708 0.655
0.800 0.795 0.740 0.755
0.740 0.748 0.574 0.681
0.635 0.667 0.618 0.637
0.748 0.700 0.630 0.638
0.713 0.678 0.585 0.635
       15-30   1978  0.709 0.601 0.701 0.724  n.s.
              1979  0.722 0.616 0.676 0.702  n.s.
              1980  0.797 0.709 0.704 0.721  n.s.
0.696 0.722 0.700 0.655
0.638 0.708 0.678 0.649
0.748 0.752 0.712 0.740
0.646 0.644 0.633 0.644
0.613 0.536 0.527 0.462
0.683 0.703 0.712 0.689
       30-46   1978  0.619 0.468 0.483 0.553  n.s.
              1979  0.694 0.493 0.549 0.610  n.s.
0.513 0.641 0.618 0.677
0.492 0.606 0.481 0.668
0.651 0.610 0.604 0.503
0.581 0.568 0.591 0.591
       61-76   1978  0.651 0.514 0.517 0.481  n.s.
              1979  0.640 0.430 0.561 0.576  n.s.
0.528 0.562 0.416 0.691
0.516 0.535 0.423 0.644
0.545 0.783 0.599 0.698
0.565 0.555 0.520 0.537
                                          (continued)

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TABLE 1.  (continued)
Soil Type
Blount silt
Ana-
lyte
Ca
I
Depth Year
cm
0-15 1978
1979
1980
15-30 1978
1979
1980
30- A b 1978
1979
61-76 1978
1979
Ck

1.37
1.06
1.28
1.19
0.83
0.62
0.71
0.34
0.97
0.77
1/4
Max

1.37
1.24
1.11
1.21
0.64
0.58
0.80
0.31
1.40
0.98
1/2
Max

1.26
0.85
0.98
0.64
0.46
0.34
0.37
0.45
1.28
0.86
loam
Max LSD

1.54 n.s.
1.26 n.s.
1.17 n.s.
0.84 n.s.
0.66 n.s.
0.41 n.s.
0.78 n.s.
0.67 n.s.
0.90 n.s.
0.84 n.s.
Elliott silt loam
Ck

•
0.72
0.68
0.82
1.03
1.10
0.69
1.07
0.37
0.92
2.22
1/4
Max

0.88
0.68
0.71
0.69
0.52
0.50
0.85
0.40
0.57
0.82
1/2
Max

1.11
0.84
0.77
0.91
1.05
0.69
1.37
1.18
2.20
2.09
Max

0.90
0.66
0.80
1.31
1.45
0.46
2.09
1.24
0.60
0.40
Plainfleld
Ck

0.55
0.54
0.65
0.62
0.52
0.40
0.40
0.25
0.30
0.22
1/4
Max

0.72
0.64
0.71
0.90
0.38
0.51
0.44
0.37
0.35
0.26
loamy
1/2
Max

0.56
0.44
0.47
0.77
0.45
0.42
0.47
0.37
0.37
0.24
sand
Max

0.93
0.59
0.71
0.51
0.38
0.35
0.30
0.32
0.41
0.30
                                               (continued)

-------
TABLE 1.  (continued)
                                                         Soil Type
Blount silt Loam
Ana-
lyte
Depth Year
cm
Ck
1/4
Max
1/2 Max LSD
Max
Elliott silt loam
Ck 1/4
Max
1/2 Max
Max
Plainfleld loamy sand
Ck
1/4
Max
1/2
Max
Max
        0-15   1978  0.894 0.915 0.833 0.949   n.s.
               1979  0.750 0.877 0.606 0.786   n.s.
               1980  0.647 0.647 0.645 0.712   n.s.
0.589 0.711 0.785 0.680
0.532 0.518 0.595 0.545
0.608 0.690 0.608 0.571
0.359 0.449 0.304 0.534
0.341 0.396 0.246 0.336
0.458 0.520 0.367 0.391
        15-30  1978  0.765 0.802 0.660 0.671   n.s.
               1979  0.565 0.598 0.490 0.500   n.s.
               1980  0.527 0.557 0.506 0.510   n.s.
0.735 0.555 0.709 0.884
0.706 0.418 0.737 0.901
0.574 0.442 0.623 0.679
0.341 0.404 0.'339 0.252
0.249 0.167 0.179 0.149
0.234 0.283 0.213 0.201
        30-46  1978  0.627 0.783 0.557 0.748  n.s.
               1979  0.527 0.721 0.750 0.782  n.s.
0.692 0.710 1.10  1.28
0.690 0.628 1.17  0.984
0.195 0.192 0.167 0.148
0.135 0.128 0.153 0.153
        61-76  1978  0.926 1.08  1.19  0.982   n.s.
               1979  0.727 1.04  0.876 0.796   n.s.
0.91  0.89  1.09  0.770
0.966 0.842 1.01  0.663
0.133 0.162 0.156 0.238
0.122 0.119 0.095 0.119
                                               (continued)

-------
TABLE 1.  (continued)
Soil Type
Ana- Depth Year
lyte Clu
Fe 0-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979

Ck

2.37
2.51
2.72
3.11
2.55
2.91
2.82
2.99
4.49
4.59
Blount
1/4
Max

2.56
2.49
2.55
3.56
3.32
3.05
4.18
5.74
4.69
4.20
silt
1/2
Max

2.49
2.07
2.28
3.23
2.66
2.69
3.46
4.10
4.76
4.03
loam
Max

2.38
2.38
2.24
2.69
2.26
2.88
3.12
3.51
5.32
4.25
Elliott silt loam
LSD

n.s.
0.29*
n.s.
n.s.
n.s.
n.s.
n.s.
1.66*
n.s.
n.s.
Ck

2.22
2.22
2.30
3.23
2.68
2.81
3.64
3.97
4.66
4.80
1/4
Max

2.54
2.56
2.43
3.01
2.60
2.50
2.71
4.27
7.33
5.97
1/2
Max

2.60
2.51
2.17
3.24
2.65
2.92
3.03
3.88
4.34
3.90
Max

2.87
2.76
2.63
3.67
2.91
3.08
1.90
3.19
4.34
4.18
Plainfield
Ck

1.05
1.37
1.59
1.39
1.14
1.05
0.78
0.66
0.56
0.66
1/4
Max

1.19
1.51
1.66
1.49
0.63
1.42
0.55
0.77
1.06
0.66
loamy
1/2
Max

1.14
1.32
1.10
1-.34
0.81
1.47
1.27
0.76
0.80
0.57
sand
Max

1.96
1.60
1.87
1.15
0.63
1.14
0.60
0.94
1.28
0.66
                                                (continued)

-------
TABLE 1.  (continued)
Soil Type
Blount silt
Ana- Depth Year Ck
lyte cm
1/4
Max
1/2
Max
loam
Max
Elliott silt loam
LSD
Ck
1/4
Max
1/2
Max
Max
Plainfield
Ck
1/4
Max
loamy sand
1/2
Max
Max


Mn 0-15 1978 847
1979 833
5 1980 898
15-30 1978 921
1979 889
1980 1020
30-46 1978 760
1979 698
61-76 1978 747
1979 688
791
752
839
716
682
787
615
857
627
661
924
827
1000
791
681
732
577
502
723
686
887
848
860
876
749
842
594
481
670
778
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
1010
796
868
835
709
886
633
586
718
678
1100
833
958
926
904
951
869
781
828
854
787
729
661
812
759
930
751
447
749
637
912
818
728
1243
898
1080
860
781
495
618
323
349
502
297
320
308
246
167
137
153
357
329
402
301
190
271
183
151
150
146
278
243
237
255
209
283
228
158
150
97
447
308
410
235
184
277
183
183
265
159
                                                (continued)

-------
     TABLE 1.   (continued)
oo

Soil Type
Blount silt
Ana- Depth Year
lyte cm
Ck
I/A
Max
1/2
Max
loam
Max LSD
Elliott silt loam
Ck
1/4
Max
1/2
Max
Max
Plain field
Ck
1/4
Max
loamy sand
1/2
Max
Max

	 — mg/kg 	 — - - - - 	 	
Zn 0-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979
81
89
87
75
83
76
90
74
92
96
180
134
159
82
89
85
109
111
97
108
267
178
240
74
83
73
96
90
105
96
379 37**
236 67**
313 85**
74 n.s.
67 n.s.
91 n.s.
84 n.s.
83 n.s.
108 n.s.
104 n.s.
79
80
84
67
78
76
76
86
92
93
187
144
187
77
94
83
84
99
158
154
263
196
184
75
83
90
84
89
82
93
673
428
551
200
190
114
142
110
125
102
35
51
58
24
37
30
18
19
21
18
137
143
129
64
37
52
15
19
12
17
199
203
139
159
103
69
26
42
13
18
804
442
639
145
118
129
30
28
22
24
                                                    (continued)

-------
TABLE 1.  (continued)
 Ana-  Depth  Year
 lyte   cm
Ck
                                                        Soil Type
                          Blount silt loam
1/4
Max
1/2
Max
Max   LSD
                                  Elliott silt loam
Ck
I/A
Max
1/2
Max
Max
Plainfield loamy sand
Ck   1/4   1/2   Max
     Max   Max
                                                        -mg/kg-
 Cd     0-15   1978   0.62  4.65  9.41 13.5  1.37**
              1979   0.82  2.84  5.40  7.92 3.00**
              1980   0.54  3.95  7.86 11.5  2.90**
                                0.65  4.84  8.96 33.5
                                0.76  2.86  6.36 18.4
                                0.69  5.15  6.18 25.3
                                                       0.53  3.97  7.15 41.4
                                                       0.64  4.69  7.53 22.7
                                                       0.87  4.20  5.29 32.2
       15-30  1978  <0.25 <0.25 <0.25
              1979   0.30 <0.25  0.43
              1980  <0.25 <0.25 <0.25
                 0.31 n.s.      <0.25  0.52 <0.25  5.16
                 0.50 n.s.      <0.25  0.82  0.53  4.53
                 0.54 n.s.      <0.25 <0.25  0.47  1.18
                                                      <0.25  1.69  6'.06  5.32
                                                       0.34  1.13  3.78  4.83
                                                      <0.25  1.17  2.05  4.36
       30-46  1978  <0.25 <0.25  0.32 <0.25 n.s.
              1979  <0.25 <0.25 <0.25  0.30 n.s.
                               <0.25  0.40 <0.25  3.22
                                0.30  0.31 <0.25  1.54
                                                      <0.25  0.37  0.49  0.62
                                                      <0.25 <0.25  1.18  0.45
       61-76  1978  <0.25  0.36 <0.25 <0.25 n.s.
              1979  <0.25 <0.25 <0.25  0.32 n.s.
                               <0.25  0.28 <0.25  1.48
                               <0.25  0.34  0.39  0.52
                                                      <0.25 <0.25 <0.25 <0.25
                                                      <0.25 <0.25 <0.25 <0.25
                                               (continued)

-------
    TABLE 1.   (continued)
NJ
O
Soil Type
Ana- Depth Year
lyte cm

Ck
Blount
1/4
Max
silt
1/2
Max
loam
Max
Elliott silt loam
LSD
Ck
1/4
Max
1/2
Max
Max
Plainfield
Ck
1/4
Max
loamy sand
1/2
Max
Max


Cu 0-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979
26
22
28
18
19
23
25
22
24
36
44
36
45
31
25
26
36
36
32
41
68
48
67
23
22
22
33
29
36
37
94
62
86
21
19
27
29
25
34
42
8**
20**
20**
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
26
19
26
19
19
22
29
29
32
35
44
36
51
21
26
21
27
30
53
48
68
50
54
19
23
26
30
30
25
40
214
125
184
49
54
29
45
31
36
42
12
12
21
7
9
9
6
6
9
10
33
36
37
14
19
15
14
7
76
16
50
52
36
'36
31
23
9
10
19
47
245
150
224
49
42
38
11
6
4
10
                                                    (continued)

-------
TABLE 1.  (continued)
Soil Type
Ana- Depth Year
lyte cm

Ck
Blount
1/4
Max
silt
1/2
Max
loam
Max
Elliott silt loam
LSD
Ck
1/4
Max
1/2
Max
Max
Plainfield loamy
Ck
1/4
Max
1/2
Max
sand
Max
	 	 	 -mol\to 	 	 	 	 	 _

Ni 0-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979
30
26
36
21
22
29
22
24
33
32
32
27
38
25
24
31
35
38
37
48
35
27
39
18
19
24
29
31
40
43
40
28
43
19
18
26
22
27
42
40
7**
n.s.
4*
n.s.
n.s.
n.s.
9*
n. s.
n.s.
n.s.
29
28
31
18
21
24
29
33
39
41
30
23
36
15
19
24
22
29
48
39
36
27
41
17
23
26
34
40
37
44
75
51
68
30
32
40
30
30
27
31
14
16
20
11
14
13
9
9
3
9
18
19
24
18
7
16
8
7
6
9
17
17
17
•14
14
15
13
11
5
9
73
39
63
16
14
20
8
10
11
9
                                               (continued)

-------
     TABLE  1.   (continued)
to
ro
Soil Type
Blount silt
Ana- Depth Year
lyte cm
Ck

1/4
Max
1/2
Max
loam
Max

Elliott silt loam
LSD

Ck

1/4
Max
1/2
Max
Max

Plain field loamy sand
Ck

1/4
Max
1/2 Max
Max

— — — — ____ — _ -ing/ Kg — - — — - —
Cr 0-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979
54
59
40
47
39
40
44
40
54
41
95
89
63
51
43
45
62
56
63
54
145
104
87
48
40
44
62
52
59
56
207
126
121
44
41
48
49
52
55
48
30**
36**
26**
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
50
60
37
45
45
36
51
52
58
57
90
81
71
46
42
39
43
46
54
46
155
110
91
48
48
36
70
59
61
64
432
193
220
94
87
38
70
60
69
40
29
38
25
24
26
17
10
9
14
8
55
85
52
43
22
27
15
8
14
3
90 522
105 234
39 352
83 61
55 65
24 49
12 7
18 19
13 20
3 6
                                                     (continued)

-------
             TABLE 1.   (continued)
       hO
       UO

Soil Type
Ana- Depth Year
lyte cm

Ck
Blount
1/4
Max
silt
1/2
Max
loam
Max
Elliott silt loam
LSD
Ck
1/4
Max
1/2
Max
Max
Plainfleld loamy .sand
Ck
1/4
Max
1/2
Max
Max


Pb 0-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979
26
27
34
19
23
28
19
21
21
23
38
38
47
18
21
24
21
28
21
24
58
51
67
19
19
18
18
23
21
20
78
61
81
20
18
24
17
22
22
23
9**
16**
21**
n.s.
n.s.
6*
n.s.
n.s.
n.s.
n.s.
25
23
28
19
19
20
18
22
19
20
37
41
51
21
27
20
21
26
32
30
56
53
48
23
22
24
18
22
18
20
158
104
145
41
36
38
28
25
31
22
13
17
30
9
11
10
7
7
7
6
31
40
36
24
18
14
7
7
11
6
48
51
37
' 36
31
39
8
11
5
6
177
124
158
27
30
26
7
12
10
6
                                                            (continued)
(m  I

-------
TABLE 1.  (continued)



	

Blount silt
Ana- Depth
lyte cm

PH 0-15

I
15-30


30-46

61-76

Year

1978
1979
1980
1978
1979
1980
1978
1979
1978
1979
Ck

7.5
7.6
7.4
6.9
6.9
6.8
6.4
6.2
5.9
5.8
1/4
Max

7.6
7.5
7.3
6.9
6.7
6.9
6.5
5.5
7.0
7.2
1/2
Max

7.3
7.1
7.1
6.1
5.9
5.5
5.4
5.6
6.7
6.7

Loam
Max

7.1
7.1
7.0
6.2
6.0
5.9
5.8
5.6
5.7
6.3

Soil
Type
Elliott silt loam
LSD

0.3*
n.s.
0.2*
n.s.
n.s.
1.1**
n.s.
n.s.
n.s.
n.s.
Ck

7.2
7.0
7.0
7.4
7.4
7.3
6.8
6.2
6.9
7.6
1/4
Max

7.2
7.0
7.1
6.9
6.7
6.7
6.7
6.9
6.2
6.6
1/2
Max

7.4
7.2
7.2
6.9
7.2
7.0
6.7
6.9
7.3
7.7
Max

5.8
6.1
6.1
6.4
7.1
6.8
7.0
5.3
4.9
5.0
Plainfield
Ck

7.4
7.4
7.3
7.3
7.3
7.5
7.0
7.2
6.8
7.2
1/4
Max

7.7
7.4
7.4
7.6
7.6
7.2
7.5
7.3
6.8
7.4
loamy sand
1/2
Max

7.0
7.1
7.1
7.1
7-5
7.4
6.8
6.8
6.6
6.7
Max

5.9
6.2
6.1
6.0
7.4
6.3
5.8
6.5
6.5
7.1
a/  Blount-silt loam control'and sludge-treated plots were replicated three times.
    on other soil types were not replicated within a series.
*,**  Significantly different at P<0.05 and P<0.01, respectively.
                                                                                    Sludge treatments

-------
low and highly variable as expected.  Because Plaiafield is so droughty,
corn is seldom grown on it in Illinois.  Sludge did not improve the pro-
ductivity of the sandy soil beyond its fertilizer value.  Furthermore,
there was no indication that sludge improved yields from Plainfield compared
to those obtained with inorganic commercial fertilizers.

   Other than effects on Mn, Cd, and Zn concentrations, there were no ob-
vious changes in plant composition attributable to previous applications
of sludge (Table 3).   Residual effects of sludge applications were mani-
fested by lower leaf- and grain-Mn; higher leaf-, grain- and Stover- Zn;
and higher leaf-and Stover-Cd concentrations with higher sludge loading
rates.  Where plots had not received sludge for 4 years, since 1973, concen-
trations of Cd in grain were not significantly different than those in
grain from control plots, regardless of" soil type.  It is noteworthy that
even though Plainfield had a very low (3 to 4 meq/100 g) and Elliott a rather
high (14 to 15 meq/100 g) cation exchange capacity, Cd and Zn contents in
corn tissues were not significantly different.  This observation was even
more phenomenal in view of the 2-fold or higher yields of grain and stover
from Elliott plots.  These data suggest that the diluting effect generally
expected as a result of better growth did not occur where the metals were
supplied as constituents of sludge.

   Four years after sludge applications were terminated on maximum-treated
Blount plots, concentrations of Cd and Zn in grain from these plots were
not different from those in grain from control plots.  But 4 years after
sludge applications were terminated on maximum-treated Elliott and Plain-
field, levels of Cd and Zn in grain were significantly higher than those in
grain from control plots even though a substantial reduction in concentra-
tions had occurred.  At the time sludge applications were terminated, maximum-
treated Blount contained 12.3+3.6 and 327+9.3 mg/kg of Cd and Zn, whereas,
when maximum sludge applications were terminated on Elliott and Plainfield,
they contained 26+7.5 and 32+9.3 mg/kg of Cd, respectively, in their surface
layers.  Concentrations of Zn in these two sludge-treated soil types were
409+17.3 and 623+181, respectively.  Thus, it appears that where higher
amounts of sludge borne Cd and Zn accumulated in soils, a longer period
without further sludge applications will be required to reduce uptake of
these metals to background levels by the particular hybrid used in the study.

Summary and Conclusions

   The results of this study showed that organic matter accumulated in soils
by sludge application remained at fairly stable contents after the first
year following the termination of applications.  It nevertheless provided
adequate amounts of N for acceptable corn yields.for 3'years beyond the
last year sludge was applied.

   After sludge applications were terminated, Cd and Zn concentrations in
corn plant tissues decreased with time.  Relatively speaking, concentrations
of Cd in corn grain decreased with time more rapidly than did those of Zn.
The length of time after terminating sludge applications required for Cd
and Zn to reach background levels of these metals in crop tissues depends
on total amounts of sludge-borne metals accumulated in soils.  The length


                                        25

-------
     TABLE 2.  CORN GRAIN AND STOVER YIELDS FROM LYSIMETER PLOTS OF THREE SOIL TYPES, WITH AND WITHOUT
               SEWAGE SLUDGE IRRIGATIONS.   ALL THREE TYPES WERE REPRESENTED IN A NORTH  SERIES.
10



Blount
Year Ck 1/4
Max
1978 2.81 3.
1979 7.96 8.
1980 6.74 6.
I 1978 4.20 4.
1979 7.69 8.
1980 6.65 6.
24
73
67
76
14
80

silt
1/2
Max
4.18
9.22
6.29
6.55
9.11
7.16

loam a
Max
5.71
9.40
7.02
5.43
9.53
7.73
Soil Type
Elliott silt loam D
LSD Ck 1/4 1/2 Max
Max Max
Grain Yields
mt/ha (15.5% moisture)
1.69** 2.81 3.40 2.45 3.99
n.a. 8.03 9.40 9.53 7.51
n.s. 5.22 4.37 4.63 2.70
Stover Yields
mt/ha (dry weight)
1.44* 5.00 6.28 4.78 5,35
n.s. 8.86 9.42 10.4 10.7
n.s. 7.35 9.70 8.92 9.68




Plainfield
Ck
0.
4.
2.
1.
4.
3.
494
44
54
06
07
28

loamy
1/4 1/2
Max Max
0.
3.
1.
1.
4.
2.
753
25
82
52
03
76
1.24
3.60
2.24
2.12
4.18
5.03


sand D
Max
0.
2.
1.
6.
4.
3.
565
92
04
48
79
58
           ^-/Sludge application stopped after 1973.
            -/Sludge application stopped after 1973 except  on Elliott  and Plainfield maximum sludge plots
              applications continued through 1976.

-------
   TABLE  3.  CONCENTRATIONS  OF  MACRO-  AND MINOR-  ELEMENTS  IN CORN LEAF,  GRAIN,  STOVER GROWN ON
              BLOUNT SILT LOAM,  ELLIOTT SILT LOAM, AND PLAINFIELD  LOAMY SAND LYSIMETER PLOTS, WITH AND WITH-
              OUT DIGESTED SLUDGE  IRRIGATIONS.a/b/
Ni

param- Year
eter
•
N 1978
1979
1980

1978
1979
1980

1978
1979
1980


Ck


1.76
2.78
2.53

0.39
0.67
0.80

0.90
1.49
1.50

Blount
1/4
Max


1.80
2.53
2.56

0.34
0.59
0.87

1.02
1.49
1.69

silt
1/2
Max


1.91
2.50
2:26

0.35
0.56
0.88

1.14
1.54
1.55

loam
Max LSD


2.20 0.30**
2.67 n.s.
2.58 n.s.

0.37 n.s.
0.71 n.s.
0.95 n.s.

1.26 0.24*
1.59 n.s.
1.56 n.s.
Soil Type


Elliott silt loam
Ck 1/4
Max

%_
Leaf
1.96 1.73
2.83 2.75
2.54 2.48
Stover
0.52 0.40
0.81 0.61
0.79 0.80
Grain
1.27 1.02
1.44 1.59
1.58 1.82
1/2
Max


1.66
2.37
2.39

0.52
0.61
1.38

1.12
1.72
1.73
Max


3.31
2.82
2.91

1.18
1.05
1.53

1.86
1.73
2.25


Plainfield
Ck


1.36
2.16
2.22

0.64
0.71
0.69

1.34
1.54
1.42
1/4
Max


1.38
2.35
2.41

0.33
0.79
0.60

1.43
1.80
1.75

loamy
1/2
Max


1.59
2.72,
2.02'

0.48
0.85
0.82

1.30
1.71
1.84

sand
Max


2.38
3.04
2.35

1.12
1.42
1.40

1.73
1.81
1.31
                                             (continued)

-------
      TABLE 3.  (continued)
     Param-
      eter
Year
                                                         Soil Type
                            Dlount silt loam
                                         Elliott sill loam
Ck
1/4
Max
1/2
Max
Max   LSD
1/4
Max
1/2
Max
Max
PlainEield loamy sand
Ck   1/4   1/2   Max
     Max   Max
                                                           Leaf
ho
GO
1978  0.315 0.285 0.226 0.238 0.032** 0.252 0.278 0.239 0.351
1979  0.277 0.256 0.243 0.252  n.s.   0.281 0.262 0.250 0.308
1980  0.264 0.263 0.270 0.267  n.s.   0.310 0.278 0.300 0.333
                                                        0.385 0.374 0.335 0.389
                                                        0.300 0.308 0.282 0.347
                                                        0.325 0.298 0.255 0.288
                1978  0.163 0.127 0.117 0.071  n.s.
                1979  0.090 0.084 0.067 0.092 0.014*
                1980  0.063 0.056 0.050 0.073  n.s.
                                          Stover


                                      0.146 0.186 0.230 0.224
                                      0.108 0.0'87 0.111 0.140
                                      0.058 0.110 0.125 0.171

                                           Grain
                                                        0.375 0.351 0.351 0.364
                                                        0.134 0.117 0.063 0.245
                                                        0.125 0.039 0.066 0.079
                1978  0.312 0.303 0.286 0.283  n.s.
                1979  0.338 0.303 0.263 0.280 0.043*
                1980  0.315 0.330 0.349 0.339 0.024*
                                      0.302 0.280 0.324 0.369
                                      0.358 0.300 0.299 0.285
                                      0.344 0.352 0.384 0.346
                                                        0.354 0.347 0.375 0.371
                                                        0.316 0.366 0.274 0.288
                                                        0.342 0.369 0.361 0.370
                                                    (continued)

-------
     TABLE 3.   (continued)
VO
	 _ _

Param- Year
eter


Ca 1978
1979
1980


1978
1979
1980


1978
1979
1980





0
0
0


0
0
0







Ck


.593
.770
.681



.427
.488
.314



61
52
54
— — — — — —
Blount
1/4
Max
•

0.685 0
0.794 0
0.730 0



0.425 0
0.489 0
0.335 0



45
60
51



silt loam
1/2 Max
Max
•

.751 0.
.800 0.
.687 0.



.409 0.
.507 0.
.279 0.



48
54
48


739
760
674



470
463
234



42
62
57
LSD


n.s.
n.s.
n.s.



n.s.
n.s.
n.s.



n.s.
n.s.
n.s.
Soil Type
Elliott silt loam
Ck 1/4 1/2 Max
Max Max
Leaf

	 _
0.581 0.618 0.737 0.618
0.756 0.767 0.829 0.773
0.586 0.666 0.720 0.669
Stover


0.355 0.369 0.336 0.538
0.426 0.388 0.473 0.438
0.280 0.204 0.264 0.256
Grain
i\.
— 	 ing/ Kg- 	 	 - —
56 64 44 45
52 63 48 39
37 51 60 42




Plainfleld loamy sand
Ck


0.567
0.643
0.534



0.394
0.466
0.224



57
39
46
1/4
Max


0.589
0.610
0.571



0.394
0.465
0.243



52
57
42
1/2
Max


0.690
0.644
0.614
•


0.358
0.510
0.337



50
42
51
Max
•

0.687
0.864
0.612



0.350
0.507
0.238



35
45
60
                                                    (continued)

-------
    TABLE  3.   (continued)
     Param-
     eter
                Year
                                                         Soil Type
                            Blount silt loam
                                         Elliott silt  loam
        Ck
1/4
Max
1/2
Max
Max   LSD
Ck
I/A
Max
1/2
Max
Max
Plainfield loamy sand
Ck   1/4   1/2   Max
     Max   Max
      Mg
CO
o
1978  0.288 0.308 0.307 0.266  n.s.
1979  0.338 0.336 0.347 0.315  n.s.
1980  0.380 0.380 0.378 0.343  n.s.
                1978  0.263 0.251 0.239 0.269  n.s.
                1979  0.242 0.227 0.225 0.224  n.s.
                1980  0.186 0.183 0.189 0.143  n.s.
                              Leaf


                         0.286 0.296 0.185 0.225
                         0.359 0.361 0.342 0.316
                         0.368 0.400 0.380 0.286

                             Stover


                         0.254 0.252 0.275 0.283
                         0.224 0.209 0.244 0.222
                         0.167 0.153 0.197 0.158

                              Grain
                                             0.387  0.364  0.371  0.302
                                             0.368  0.398  0.388  0.210
                                             0.324  0.319  0.357  0.330
                                                               0.330 0.307 0.275 0.241
                                                               0.290 0.279 0.306 0.339
                                                               0.157 0.153 0.227 0.168
                1978  0.117 0.116 0.119 0.119  n.s.
                1979  0.189 0.115 0.107 0.150  n.s.
                1980  0.140 0.141 0.151 0.147  n.s.
                                      0.124 0.111  0.128  0.138
                                      0.147 0.122  0.120  0.108
                                      0.144 0.147  0.166  0.147
                                                   0.137 0.131  0.145  0.153
                                                   0.127 0.170  0.120  0.130
                                                   0.147 0.167  0.152  0.160
                                                    (continued)

-------
TABLE 3.  (continued)

Soil Type
Blount silt
Pararn- Year
eter
.
/
Fe 1978
1979
1980

1978
1979
1980

1978
1979
1980
Ck



94
135
107

227
262
174

11
19
22
1/4
Max



91
116
96

202
256
182

10
16
24
1/2
Max



104
97
107

239
198
153

11
16
22
loam
Max LSD



100 n.s.
115 n.s.
94 n.s.

139 n.s.
279 n.s.
130 n.s.

14 n.s.
17 n.s.
24 n.s.
Elliott silt loam
Ck 1/4
Max


Leaf
91 85
114 124
109 100
Stover
112 190
194 160
145 118
Grain
14 11
26 18
23 25
1/2
•Max



73
98
109

193
219
149

12
17
26
Max



103
116
104

103
160
137

18
19
29
Plainfield loamy sand
Ck


•
97
118
102

186
154
175

12
17
17
1/4
Max



96
116
87

223
262
183

12
22
23
1/2
Max



100
116
92,

100
282
172

23
16
29
Max

•

96
102
104

100
250
112

21
17
19
                                                (continued)

-------
     TABLE 3.   (continued)
N)
Soil Type
Param- Year
eter


Mn 1978
1979
1980

1978
1979
1980

1978
1979
1980

Ck



28
41
40

41
37
29

4.0
6.8
7.5
Blount
1/4
Max



30
31
30

34
39
28

3.6
5.3
7.2
silt
1/2
Max



28
36
38

39
40
29

4.0
4.7
7.7
loam
Max LSD



26 n.s.
28 9**
29 8*

45 n.s.
38 n.s.
26 n.s.

4.0 n.s.
4.6 1.4**
7.2 n.s.
Elliott silt loam
Ck



29
42
38

29
36
26

4.
7.
7.
1/4
Max


Leaf
26
42
38
Stover
32
27
24
Grain
8 3.3
4 5.5
7 8.0
1/2
Max



15
29
29

23
40
23

3.2
4.7
7.6
Max



40
41
27

50
26
22

3.5
3.2
5.9
Plainfield
Ck



24
37
32

28
24
22

3.8
5.3
6.1
1/4
Max



29
35
31

27
32
21

3.2
7.5
7.4
loamy
1/2
Max



19
25
22
•
20
26
19

3.9
4.0
6.1
sand
Max



56
55
29

36
23
14

4.0
3.4
3.9
                                                     (continued)

-------
     TABLE 3.   (continued)
co

Soil Type
Blount silt
Param- Year
eter


Zn 1978
1979
1980

1978
1979
1980

1978
1979
1980
Ck



20
32
25

51
22
23

28
19
21
1/4
Max



26
46
34

35
30
32

26
19
23
1/2
Max



25
50
43

34
33
41

26
18
25
loam
Max



34
62
43

32
50
49

28
20
28
Elliott silt loam
LSD



8**
16**
12**

n.s.
9**
18**

n.s.
n.s.
5**
Ck 1/4
Max


Leaf
19 30
28 72
26 42
Stover
28 45
17 40
14 40
Grain
26 28
19 20
21 26
1/2
Max



27
67
56

53
46
62

31
21
33
Max



279
212
148

253
210
187

53
27
36
Plainfield loamy sand
Ck



20
31
28

70
12
36

34
15
21
1/4
Max



45
73
42

112
53
39

37
24
25
1/2
Max



51
100
52

98
88
69

38
22
28
Max



246
382
124

326
244
178

45
29
34
                                                     (continued)

-------
     TABLE 3.   (continued)
u>

Soil Type
Blount silt
Pa ram- Year
eter
Cd 1978
1979
1980
1978
1979
1980
1978
1979
1980
Ck

0.4
0.3
0.2
0.6
0.4
0.4
<0.06
<0.06
<0.06
1/4
Max

1.1
0.8
0.4
0.7
1.1
1.0
<0.06
<0.06
0.08
1/2
Max

1.9
1.3
1.0
1.5
1.9
1.5
<0.06
<0.06
0.09
loam
Max

3.1
2.1
1.0
1.8
2.8
2.0
<0.06
*0.06
<0.06
Elliott silt loam
LSD

0.4**
0.3**
0.6**
0.7**
0.6**
0.4**
n.s.
n.s.
n.s.
Ck 1/4
Max

	 ing/ kg —
Leaf
0.4 1.5
0.4 1.1
0.2 0.6
Stover
0.4 0.7
0.4 1.3
0.4 0.8
Grain
<0.06 <0.06
0.09 <0.06
<0.06 <0.06
1/2
Max

2.1
2.0
1.2
1.5
2.3
2.4
<0.06
<0.06
<0.06
Max

21.8
19.3
9.7
25.0
23.2
16.3
0.66
0.34
0.40
Plainfield loamy sand
Ck

0.3
0.4
0.3
0.7
0.4
0.8
<0.06
<0.06
<0.06
1/4 1/2
Max Max

1.1 1.2
1.6 2.2
0.8 1.2
1.3 1.3
2.1 3.6
1.3 1.7
0.09 <0.06
<0.06 <0.06
0.08 <0.06
Max

11.4
17.3
3.8
13.3
15.9
8.2
0.28
0.18
0.28
                                                     (continued)

-------
     TABLE 3.  (continued)
u>
Cn

Soil Type
Blount silt
?aram- *ear
eter


Cu 1978
1979
1980

1978
1979
1980

1978
1979
1980
Ck



3.4
8.3
8.5

1.6
6.3
5.9

1.4
2.5
2.0
1/4
Max



3.8
8.8
8.6

1.5
6.3
6.3

1.3
2.4
2.1
1/2
Max



3.9
8.6
8.9

1.8
6.2
6.5

1.5
2.2
2.1
loam
Max LSD



4.5 0.5**
8.8 n.s.
8.7 n.s.

2.3 n.s.
7.3 n.s.
6.0 n.s.

1.6 n.s.
2.2 n.s.
2.1 n.s.
Elliott silt loam
Ck 1/4
Max


Leaf
3.7 4.0
8.6 9.1
7.8 7.7
Stover
1.8 1.7
5.9 6.1
4.7 5.8
Grain
1.4 2.5
2.6 2.0
1.9 1.9
1/2
Max



4.4
8.4
8.7

2.0
11.7
6.6

1.5
2.7
2.1
Max



9.4
10.3
11.2

3.5
6.9
6.3

2.1
1.8
1.7
FlainCield loamy sand
Ck



2.7
5.8
5.6

1.8
4.9
3.8

1.5
1.8
1.6
1/4
Max



3.1
7.5
6.0

2.6
6.0
5.2

1.6
3.0
1.9
1/2
Max



3.2
8.4
5.9

2.0
7.2
5.7

1.8
2.3
1.8
Max

•

5.9
9.9
7.1

4.1
7.7
6.2

1.9
1.9
1.8
                                                     (Continued)

-------
     TABLE 3.  (continued)
10
Soil Type
Blount silt
Pa ram- Year
eter


Ni 1978
1979
1980

1978
1979
1980

1978
1979
1980
Ck



<0.6
<0.6
<0.6

0.7
<0.6
<0.6

0.6
<0.6
<0.6
1/4
Max



<0.6
<0.6
<0.6

<0.6
<0.6
<0.6

<0.6
<0.6
0.6
1/2
Max



<0.6
<0.6
<0.6

<0.6
0.9
<0.6

<0.6
<0.6
<0.6
loam
Max LSD



<0.6 n.s.
<0.6 n.s.
<0.6 n.s.

<0.6 n.s.
<0.6 n.s.
<0.6 n.s.

<0.6 n.s.
<0.6 n.s.
<0.6 n.s.
Elliott
Ck 1/4
Max


Leaf
<0.6 <0.6
<0.6 <0.6
<0.6 <0.6
Stover
<0.6 <0.6
<0.6 <0.6
0.7 <0.6
Grain
<0.6 0.9
<0.6 <0.6
<0.6 0.8
silt loam
1/2
Max



<0.6
<0.6
<0.6

0.7
<0.6
0.7

0.7

-------
     TABLE 3.   (continued)
10
Soil Type
Blount silt
Parara- Year
eter


Cr 1978
1979
1980

1978
1979
1980

1978
1979
1980
Ck



<0.6
<0.6
<0.1

0.9
1.3
0.7

0.2
0.2
<0. 1
1/4
Max



<0.6
<0.6
0.2

0.9
1.0
0.8

0.1
0.2
<0. 1
1/2
Max



<0.6
<0.6
<0.1

1.0
1.8
0.9

0.2
0.2
<0. 1
loam
Max



<0.6
<0.6
0.2

<0.6
2.1
<0.6

0.3
0.2
<0.1

LSD



n.s.
n.s.
n.s.

n.s.
n.s.
n.s.

n.s.
n.s.
n.s.
Elliott
Ck 1/4
Max


Leaf
<0.6 <0.6
<0.6 0.7
<0. 1 <0. 1
Stover
<0.6 1.3
0.7 0.7
0.8 <0.6
Grain
0.7 <0.1
0.1 <0.1
<0.1 <0. 1
silt loam
1/2
Max



<0.6
<0.6
<0.1

<0.6
1.2
0.8

0.2
0.2
<0. 1
Max



<0.6
0.6
0.2

<0.6
1.1
1.2

0.2
<0. 1
<0. 1
Plainfield loamy sand
Ck



<0.6
0.7
0.2

<0.6
1.1
0.9

0.2
0.1
<0. 1
1/4
Max



<0.6
0.6
0.6

2.3
1.9
<0.6

0.2
0.2
<0. 1
1/2
Max



<0.6
0.8
<0.1

<0.6
3.0
0.9

0.2
0.2
0.2
Max



<0.6
<0.6
0.3

<0.6
1.2
0.9

0.2
0.3
<0. 1
                                                    (continued)

-------
     TABLE 3.  (continued)
u>
oo
Soil Type
Blount silt
Param- Year
eter


Pb 1978
1979
1980

1978
1979
1980

1978
1979
1980
Ck



1.3
1.0
<0.6

1.3
2.9
2.2

0.8
1.3
<0.6
1/4
Max



1.3
1.0
<0.6

1.4
3.3
2.7

<0.6
<0.6
<0.6
1/2
Max



1.4
1.2
<0.6

1.5
3.4
3.0

<0.6
1.7
<0.6
loam
Max LSD



1.5 n . s .
0.9 n.s.
<0.6 n.s.

1.5 n.s.
3.8 n.s.
3.3 n.s.

1.4 n.s.
<0.6 1.0*
<0.6 n.s.
Elliott
Ck 1/4
Max


Leaf
1.5 1.7
1.1 0.9
<0.6 <0.6
Stover
1.3 1.9
3.3 2.5
3.6 3.3
Grain
<0.6 <0.6
1.0 0.8
<0.6 <0.6
silt loam
1/2
Max



1.4
0.9
<0.6

2.0
2.7
1.6

0.7
<0.6
<0.6
Max



1.3
1.2
<0.6

1.2
2.5
2.3

<0.6
<0.6
<0.6
Plain field loamy sand
Ck



1.3
1.8
<0.6

3.2
3.3
4.5

<0.6
1.0
<0.6
1/4
Max



1.6
1.0
<0.6

3.4
4.3
4.6

<0.6
<0.6
<0.6
1/2
Max



1.7
1.2
<0.6

2.1
4.3
3.5

0.7
0.8
<0.6
Max



1.7
1.6
<0.6

3.0
3.0
3.2

2.4
2.1
<0.6

     aj Concentrations  were calculated on a dry weight basis.
     _b/ All  three  soil  types were represented in a north series.
     *,**  Significantly  different at  P<0.05 and P<0.01, respectively.

-------
of time is longer for higher concentrations accumulated in soils by addi-
tional years of similar incremental annual applications.  The availability
of Cd and Zn for uptake by corn decreased by about 50% during the first
year after sludge applications were suspended, then more slowly thereafter.
Concentrations of trace elements in corn plant tissues were not significantly
affected by fertilizing sludge-amended soil with inorganic sources of N nor
was the trend toward lower uptakes of Cd and'Zn with time affected.

   There was no indication that soil types influenced uptake of sludge-
borne Cd and Zn by corn nor their decreased availability with time after
sludge applications were terminated.

   Since soil pH was significantly lower in plots amended with higher amounts
of sludge, further work is needed to see how pH adjustment would affect the
uptake of Cd and Zn and the rate at which these metals are converted to un-
available forms.

   Further work is needed to see if renewed annual applications of sludge
would convert unavailable sources of Cd and Zn in soils to forms more avail-
able for uptake by plants.


RESPONSE OF CONTINUOUS CORN ON BLOUNT SILT LOAM TO REPEATED ANNUAL
APPLICATIONS OF SEWAGE SLUDGE

Introduction

   This report contains the results for the last three years of a 13-year
study involving the annual irrigation of corn with digested sewage sludge,
which was initiated in 1968.  Results obtained during the first ten years
were reported previously (Hinesly et al., 1981).

Materials and Methods

   Anaerobically digested sewage sludge from Chicago, Illinois was annually
applied by furrow irrigation of corn, growing on ridges, in replicated (four)
plots (6.1 x 12 m) of Blount silt loam soil located on the northeast Agronomy
Research Center near Elwood, Illinois.  Sludge applications of 0, 6.4, 12.7,
and 25.4 mm, randomized within blocks, were made as soon and as often as weather
conditions permitted after plants reached a height of about 15 cm.  Prior to
spring tillage operations, all plots received broadcast applications of KC1
to supply 134 kg K/ha.  Control plots were also fertilized with 268 kg N/ha
and 134 kg P/ha.

   At the beginning and end of each irrigation event samples of digested
sludge were collected, dried at 110 C for 24 hours, and ground to pass an
80-mesh screen.  Soil samples were collected each spring after the first
tillage to a depth of 76 cm using stainless steel tubes.  Soil cores were
segmented into 15.2 cm lengths, composited for a particular depth, dried
(60 C), crushed, split, and pulverized to pass a 60-mesh screen.  Sludge
and soil samples (0.1 and 0.5 g, respectively) were heated to 500°C for 24
hours, digested in concentrated HC1-HF, and dissolved in 0.1 N^ HC1 for analy-


                                       39

-------
sis by atomic absorption spectrephotometry to determine metal concentrations.

   When about 10% of the plants had tasseled, leaves  opposite and below the
primary ear shoots were collected from the two center rows of each plot,
washed in distilled water, dried at 60°C and ground in a Wiley mill to pass a
20-mesh screen.  Subsamples of grain,  hand-harvested  from the two center rows
to determine yields, were dried and ground iir the same manner as leaf samples.
Leaf and grain samples (2 g) were digested in concentrated HNO. at 90 C,  fol-
lowed by HCIO^ at 200°C, taken to dryness, dissolved  in 0.1 N. HNO_, and ana-
lyzed for metal concentrations by atomic absorption spectrophotometry.

   Phosphorus concentrations in solutions of digested materials were deter-
mined colorimetrically using a vanadomolybdate finish.  Total organic-C con-
centrations in soil were determined by the Walkley-Black procedure (Allison,
1965).  Soil cation exchange capacities (CEC) were determined by the ammonium
saturation method (Chapman, 1965) .   Total nitrogen concentrations were deter-
mined by a semimicro-Kjeldahl method (Brenner,  1965).

   The study protocol, as described in the previous report, remained un-
changed during the last three years, with one exception.  Because corn
hybrids have different inherited capacities to take up trace elements, seed
of one hybrid (Pioneer 3517) had been used each year  since the study was ini-
tiated.  But in 1980, it was learned that this hybrid would no longer be
marketed and was being replaced by the new hybrid 3541.  Toward maintaining
continuity of results, enough seed of hybrid 3517 was purchased to plant
split-plots (4 rows) for three years so that comparison of results with
those from hybrid 3541 could be made.   Hybrid 3541, planted for the first
time in 1980, will be planted on the other half of split-plots during each
of two additional years.  Knowing the relationship between the responses of
the two hybrids to sludge irrigation will permit the  evaluation of long-term
effects without the confounding effects that generally occur in such studies
when different hybrids, cultivars,  or varieties are substituted.

Results and Discussion

   Amounts of liquid digested sludge,  applied by furrow irrigation, on maximum-
treated plots, during the last three years, ranged from 12.7 to 17.8 cm and
supplied equivalent dry weight solids loading rates that ranged from 45.8
to 68.4 mt/ha (Table 4).  Appropriately lesser amounts were applied on the
same days on 1/4- and 1/2-maximum-treated plots.  Concentrations of various
chemical constituents of digested sludge are presented in Table 5, along with
total equivalent dry weights of these constituents applied during a particular
year.  On maximum-treated plots N and P were applied  at rates that were about
ten fold amounts needed to satisfy nutrient requirements of corn.  Slightly
less than half of the total N was in the readily available NH^-N form (Table
6).  At the end of the 1980 growing season 185, 1,018, 2,186, 264, 768, and
3,652 kg/ha of sludge-borne Cd, Cu, Cr, Ni, Pb, and Zn, respectively, had
been applied on maximum-sludge-treated plots during the total 13 year study.
The chemical composition of Blount silt loam was markedly changed by long-
term applications oc sewage sludge.
                                       40

-------
TABLE  $-  ANNUAL DIGESTED SLUDGE. LOADING RATES AND TOTAL ACCUMULATIONS ON
          MAXIMUM-SLUDGE-TREATED BLOUNT SILT LOAM PLOTS PLANTED TO CORN.

Liouid Sludge ADD lied (c=0
Year
1978
1979
1980
Annual
17.8
15.2
12.7
Accumulations
225.4
240.6
253.3
Dry Solids Aoplied (mt/ha)
- - Annual
68.4
62.5
45.8
Accumulations
625.0
687.5
733.3
   Concentrations of selected chemical elements at various depths  of Blount
with and without sludge are presented in Table 7.   Organic-C and total N
contents were markedly increased in the 0 to 30 cm depth of soil by sludge
applications.  However, there is no evidence that  either organic-C or N con-
tinued to accumulate to higher levels in Blount with successive years of
sludge applications.  Apparently,  for any sludge loading rate,  an  equilib-
rium between additions and decomposetional losses  of organic matter was main-
tained throughout the additional three years of study.   Some variation in
organic-C and N contents existed between years because  sludge composition
and loading rates were not always the same,  climatic factors affecting de-
compositional processes varied from year to  year,  and soil samples were not
collected each year at precisely the same time. Nevertheless,  these data
leave little doubt that organic matter contents of Blount will  not be in-
creased further without drastically increasing sludge loading rates.

   Concentrations of P were markedly increased in  the 0 to 30 cm depth of
soil by sludge applications, but those for K, Na,  Ca, Mg, Fe, and  Mn were
not affected (Table 7).  Concentrations of all the transition and heavy
metals such as Cd, Cu, Cr, Ni, Pb, and Zn were significantly increased in
the 0 to 30 cm soil depth.  There was some indication that Cd and  Zn concen-
trations may have been increased at soil depths below 30 cm.

   Limestone applications were not made during the last 3 years of the study.
As expected, maximum sludge applications caused a  significant reduction
in soil pH, when compared to values from control and 1/4-maximum-sludge-
treated plots.  Maximum sludge applications  tended to lower pH throughout
the soil profile (Table 7).

   The intent was to obtain plant populations of about  60,000 plants per
hectare, and, as can be seen in Table 8, the number of  plants did  not vary
significantly from  this value for any of the plots.'  Grain yields  were in-
creased by sludge applications during the first two years of the continua-
tion study, but during the last year treatments had no  effect on yields for
either of the two hybrids.  Due to hot, dry weather, grain yields  were sig-
nificantly less in  the last year of the study (Tables 8 and 9).  However,
stover yields were  somewhat higher during the year of lowest grain yields
and were increased by higher sludge applications,  regardless of hybrid.
Lower grain yields were perhaps due to weather conditions that adversely
affected pollination.

                                       41

-------
TABLE 5.  AVERAGE CONTENTS AND TOTAL ANNUAL AMOUNTS OF SEVERAL CONSTITUENTS OF SLUDGE APPLIED ON THE
          MAXIMUM-TREATED BLOUNT SILT LOAM PLOTS PLANTED TO CORN.  APPROPRIATELY LESSER AMOUNTS OF
          SLUDGE FROM THE SAME BATCH WERE APPLIED ON OTHER SLUDGE-TREATED PLOTS ON THE SAME DAY.
Year    Solids     Total N   P     K   Na    Ca   Mg     Fe    Mn    Cd    Cu    Cr     Ni   Pb    Zn

1978
1979
1980

— % —
3.86
4.06
3.61



2097
2227
2290



1010
1070
1022



180 87
225 101
234 132
Amounts
Annual Meai

1240 505
1160 495
1360 572
of Analytes
	 (]
is of Analyl
'kg wet we if
1530 25
1780 22
1470 21
Applied as
CD/HA Hrv u«
:es in
»hf 1- —

10.5
10.0
7.6
Sludge

62 108
67 106
56 96
Constituents of
>ioht^ 	


17.8
17.0.
15.91
Sludge


34
31
27



149
146
125

1978                3725    1790  320  154  2200  897   2720   44    19     110   192     32    60   265
1979                3956    1900  400  179  2060  879   3160   38    18     119   188     30    55   259
1980                2906    1296  297  168  1726  725   1868   26    10      70   122     20    34   159

-------
TA3LE 6.   AVERAGE CONTENTS AND TOTAL AMOUNTS OF VARIOUS FORMS OF N AND ASH
          CONTENTS OF SOLIDS ANNUALLY APPLIED AS CONSTITUENTS OF SLUDGE ON
          BLOUNT SILT LOAM.
Year          7. Ash          Total N          NH4-N
                        Annual Means of Analytes in Sludge
                                •(tag/kg wee weight)-
1978           51.9            2097            1020             7.8
1979           48.6            2227            1055            14.4
1980           51.1            2290            1071             4.4

               Amounts of Analytes Applied as Constituents of Sludge
                               —(kg/ha dry weight)-
1978            -              3725            1812             14
1979            -              3956            1874             25
1980            -              2906            1359             6
    Except  for Cd,  Cu, Ni,  and  Zn,  concentrations  of most of  the selected
 elements in leaves,  grain,  and stover, were  not consistently affected by
 sludge applications  (Table 10).   In view  of  the exceedingly  high  rates of
 sludge-borne N,  it is remarkable that N contents  in plant  tissues were never
 markedly increased,  although small,  significant increases  were observed in
 some years.  This  was also the situation  for P contents of plant  tissues.
 Except for Cd,  there was no evidence that concentrations of  trace elements in
 corn plant tissues differed between the two  hybrids used during the  last year
 of the study.  Neither  was there any indication that  trace element concentra-
 tions in plant  tissues  increased as a result of long-term  accumulations of
 sludge-borne metals  in  Blount.

 Summary and Conclusions

    During  the last three years of the 13  year study,  there was no indica-
 tion that  a phytotoxic  condition was imminent.  Except for the continual
 build up of transition  and heavy metal concentrations in sewage sludge-
 treated soil, there were few changes in the  chemical  properties of Blount.

    As was  previously reported, uptake of  Cd, Cu,  Ni,  and  Zn  were  corre-
 lated with annual sludge-borne applications  of these  metals  on Blount.

    The new corn hybrid, planted in the last  year, took up  only about 1/2
 as much Cd as the hybrid that had been used  throughout the study. Both of
 these hybrids were selected on the basis  of  their potential  to produce high
 yields when  grown in the region within which the Northeast Agronomy  Research
 Center is located.  Since  the parents of these hybrids were  unknown  outside
 the seed company, their capacities to take up metals were not known  a priori.
 However, it was obvious from results obtained during the one-year of compari-

                                        43

-------
TABLE 7.  CONCENTRATIONS OF TOTAL MACRO- AND TRACE- ELEMENTS, ORGANIC-C, AND
          pH DETERMINED FROM SOIL SAMPLES COLLECTED AT DEPTHS OF 0 TO 15, 15
          TO 30, 30 TO 46, AND 61 TO 76 cm IN BLOUNT SILT LOAM PLOTS.a/b'c'
Sludge Application Rates
Depth
Parameter on
Organic C 0-L5

15-30

30-46

61-76

N 0-15


15-30


30-46

61-76

P 0-15


15-30


30-46


61-76

Year
1978
1979
1980
1978
1979
^ X * '
1980
1978
1979

1978
1979
1978
1979
1980
1978
1979
1980
1978
1979
1978
1979
1978
1979
1980
1978
1979
1980
1978
1979
i
1978
1979
0

1.37
1.38
1.37
0.99
0.89
0.84
0.35
0.42

0.47
0.45
0.133
0.135
0.134
0.105
0.090
0.085
0.058
0.048
0.060
0.055
.081
.094
.080
.045
.045
.047
.026
.032

.032
.'035
1/4-
Max

1.68
1.87
1.66
1.31
1.18
1.07
0.44
0.47

0.44
0.41
0.151 "
0.168
0.141
0.131
0.113
0.104
0.063
0.065
0.060
0.055
.116
.144
.140
.074
.086
.078
.031
.034

.033
.036
1/2
Max
--Z-- •"-•
2.29
2.60
2.44
1.55
1.41
1.32
0.47
0.44

0.49
0.48
0.200
0.193
0.205
0.125
0.130
0.121
0.064
0.060
0.066
0.060
.198
.250
.223
.111
.109
.113
.033
.033

.034
.037
Max

2.90
3.36
3.21
2.05
1.80
1.85
0.40
0.44

0.38 .
0.45
0.279
0.290
0.257
0.204
0.173
0.16&
0.061
0.055
0.054
0.058
.333
.391
.356
.182
.194
.186
.028
.033

.028
.034
LSD

0.65**
0.27**
0.44**
0.31**
0.40**
0.36**
n.s.
n.s.
s
n.s.
n.s.
0.040**
0.057**
0.085**
0.054**
0.027**
0.031**
n.s.
n.s.
n.s.
n.s.
.061**
.046**
.048**
.023**
. 033**
.047**
n.s.
n.s.

n.s.
n.s.
(continued)
                                      44

-------
TABLE  7.  (continued)
Sludge Application Rates
Depth
Parameter cm
K 0 -15
15-30
30-46
61-76
Na 0-15
15-30
30-46
61-76
Ca 0-15
15-30
30-46
61-76
Year
1978
1979
1980
1978
1979
1980
1978
1979
1978
1979
1978
1979
1980
1978
1979
1980
1978
1979
1978
1979
1978
1979
1980
1978
1979
1980
1978
1979
1978
1979
0

1.87
1.96
2.02
1.95
1.97
1.92
2.08
2.08
2.64
2.51
0.814
0.807
0.854
0.767
0.669
0.726
0.609
0.513
0.469
0.432
0.502
0.493
0.509
0.476
0.401
0.377
0.341
0.236
0.783
1.44
1/4
Max

1.88
1.99'
1.92
1.92
1.95
1.90
1.95
1.96
2.51
2.61
0.798
0.758
0.819
0.761
0.703
0.734
0.634
0.529
0.472
0.516
0.630
0.542
0.613
0.514
0.499
0.428
0.388
0.270
0.418
0.819
1/2
Max

1.88
1.93
1.90
1.93
1.80
1.91
2.02
1.97
2.56
2.77
0.790
0.734
0.812
0.736
0.653
0.741
0.634
0.552
0.461
0.493
0.560
0.509
0.568
0.518
0.475
0.413
0.332
0.290
0.620
0.620
Max

1.82
1.89
1.86
1.87
1.93
1.88
1.96
1.94
2.43
2.69
0.778
0.736
0.838
0.748
0.706
0.736
0.684
0.515
0.519
0.503
0.640
0.552
0.618
0.517
0.500
0.425
0.343
0.263
0.598
0.556
LSD

n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
0.140*
0.024*
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
0.599*
(continued)


45





-------
TABLE 7.  (continued)
Sludge Application Rates
Depth
Parameter cm
Mg 0-15
15-30
30-46
61-76
Fe 0-15
15-30
30-46
61-76
Mn 0-15
15-30
30-46
61-76
Year
1978
1979
1980
1978
1979
1980
1978
1979
1978
1979
1978
1979
1980
1978
1979
1980
1978
1979
1978
1979
1978
1979
1980
1978
1979
1980
1978
1979
1978
1979
0

0.368
0.347
0.384
0.427
0.411
0.512
0.691
0.635
0.993
1.25
1.82
1.87
1.86
2.11
2.31
2.64
3.92
4.06
4.40
4.12
863
1210
947
805
748
625
418
442
628
667
1/4
Max

0.459
0.381
0.458
0.414
0.393
0.494
0.620
0.577
0.698
1.02 •
2.00
2.08
1.86
2.03
2.02
2.36
3.19
3.72
4.40
4.37
1065
1350
930
864
753
702
400
398
670
672
1/2
Max
._— ?_____.
0.395
0.363
0.412
0.431
0.346
0.463
0.587
0.591
0.950
0.992
2.03
2.10
1.98
2.04
2.15
2.28
3.32
3.70
4.62
4.31
— mg/kg 	
953
1280
906
786
789
747
389
412
603
751
Max

0.396
0.349
0.409
0.365
0.382
0.474
0.538
0.555
0.834
0.955
2.06
2.11
1.82
2.05
2.08
2.44
3.03
3.65
4.20
4.36
961
1050
886
948
942
664
400
414
474
667
LSD

n.s.
n. s.
n.s.
n.s.
n.s.
n. s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
                                     46

-------
TABLE 7.  (continued)
Depth
Parameter cm
Zn 0-15
15-30
30-46
61-76
Cd 0-15
15-30
30-46
61-76
Cu 0-15
15-30
30-46
61-76
Year
1978
1979
1980
1978
1979
1980
1978
1979
1978
1979
1978
1979
1980
1978
1979
1980
1978
1979
1978
1979
1978
1979
1980
1978
1979
1980
1978
1979
1978
1979

0

79
82
76
74
78
72
80
86
103
102
0.6
0.5
0.6
0.4
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
15
18
20
19
20
21
27
36
31
41
Sludge
1/4
Max

179
225 "
220
146
141
129
82
87
107
108
5.6
8.0
8.4
4.1
3.7
3.2
0.45
<0.25
<0.25
<0.25
48
50
68
34
40
38
26
33
33
43 '
Application Bates
1/2
Max

ing/ Kg
305
379
352
219
183
194
91
91
111
110
13.0
16.3
15.4
7.6
6.0
6.2
0.46
<0.25
0.30
<0.25
86
92
113
58
64
55
30
34
41
41
Max

521
578
536
362
315
317
98
1'04
109
120
24.6
28.4
25.8
15.6
13.1
13.5
0.52
<0.25
<0.25
<0.25
151
162
168
102
112
101
25
32
36
44
LSD

98**
67**
63**
48**
62**
54**
4*
n.s.
n.s.
11**
5.6**
3.1**
3.1**
2.6**
3.1**
3.2**
n.s.
n.s.
0.18*
n.s.
27**
33**
19**
25**
16**
21**
n.s.
n.s.
n.s.
n.s.
(continued)


47





-------
TABLE  7.  (continued)

Depth
PaTflTUPH^TT cm

Ni 0-15


15-30


30-46

61-76

Cr 0-15


15-30


30-46

61-76

Pb 0-15


15-30


30-46

61-76

Yaar

1970
1979
1980
1978
1979
1980
1978
1979
1978
1979
1978
1979
1980
1978
1979
1980
1978
1979
1978
1979
1978
1979
1980
1978
1979
1980
1978
1979
1978
1979

0


14
14
15
16
13
19
28
27
49
43
40
36
31
43
39
30
46
48
71
63
21
28
20
22
22
22
23
19
28
23
Sludge
1/4
Max


19.
23
24
20
14
24
23
23
43
44
120
101
78
94
68
60
47
46
74
59
43
60
52
37
35
36
27
18
26
22 '
Apolication Rates
1/2
Max
Max
— rag/kg 	
28
34
37
25
16
26
27
23
47
50
205"
171
143
135
90
74
58
45
76
64
74
99
83
53
44
48
25
18
31
21
36
47
53
32
24
38
22
22
45
45
341
270
254
222
157
114
61
45
69
62
121
144
127
83
73
80
26
19
25
21
LSD


14**
7**
7**
5**
8**
6**
n.s.
n.s.
n.s.
n.s.
51*
42**
58**
42**
31**
33**
10**
n.s.
n. s.
n.s.
23**
16**
16**
12**
15**
11**
n.s.
n.s.
n.s.
n.s.
                                  (continued)




                                     48

-------
TABLE  7.   (continued)
Depth
Parameter cm

pR 0-15


15-30


30-46

61-76

Year

1978
1979
1980
1978
1979
1980
1978
1979
1978
1979

0


5.9
6.3
6.2
5.9
5.8
5.2
4.5
4.6
6.2
7.4
Sludge
1/4
Max


6.6
• 6.8 -
6.7
6.4
6.2
5.7
4.6
4.5
5.2
6.4 •
Application Rates
1/2
Max
— Units —
6.1
6.3
6.0
5.9
5.8
5.5
4.4
4.4
5.6
6.5
Max


5.7
6.1
5.9
5.8
5.6
5.4
4.3
4.2
5.0
5.6
LSD


0.6**
0.5*
0.5*
n.s.
0.4*
n.s.
0.2**
0.2*
n.s.
0.9*
a/ Concentrations are-on a dry weight basis
j>/ Digested sludge was applied annually on replicated plots at various rates
   and control or check plots were annually treated with a relatively high
   rate of inorganic fertilizer.
cj Samples were collected from plots identified as the NW 800 series.
*,**  Significantly different at P<0.05 and P<0.01, respectively.
n.s. =.noni:signifleant F-test.
                                       49

-------
TABLE g.   PLANT POPULATIONS AND CORN GRAIN AND STOVER YIELDS FROM PLOTS
          DESIGNATED NW 800.

Year
Sludge Application Races
1/4 1/2
0 Max Max Max

LSD
                                      Population
  1978
  1979
  1980
  1980
61.5
                                —1,000 plants/ha—
                                     Pioneer 3517
56.2
61.9
59.5

54.9
63.6
61.5

57.8
59.3
62.1
Pioneer 3541
59.3
63.0
59.2

59.5
61.5
 59.2
                                    Grain Yields
  1978
  1979
  1980
  1980
  1978
  1979
  1980
  1980
                                it/ha (15.5% moisture)-
                                     Pioneer 3517
5.73
8.51
3.50

2.53

6.24
8.93
3.17

2.69

6.54
9.76
2.33
Pioneer 3541
2.76
Stover Yields
7.32
9.22
2.63

3.54

0.807**
0.820*
n.s.

n.s.


Pioneer 3517
8.02
9.41
9.86

9.84
11.1
11.5

8.35
10.6
12.5
Pioneer 3541
9.50
9.80
13.2

n.s.
n.s.
2.42**

 8.84
 9.36
10.4
11.4
1.60*
  n.s.  =  no  signirleant di±terences.
  *,  ** =  significant different at P<0.05 and P<0.01,  respectively.
                                     50

-------
TABLE 9 .   AVERAGE RAINFALL AT THE NORTHEAST AGRONOMY RESEARCH CENTER. DURING
           THE 3 GROWING SEASONS.  THE 1941 TO 1970 AVERAGE RAINFALL FOR THE
           AREA IS PRESENTED FOR COMPARISON.

Month " '
Year

L978
1979
1980
1941-1970
Average
April

9.0
11.5
5.8

9.4
Ilay

9.0
5.0
10.4

9.1
June

20.3
10.3
8.3

10.4
July
— Rainf 1 1 1
5.7
13.2 A
9.2 '

9.8
August

2.8
15.5
19.6

7.7
September

3.2
0.1
20.8

8.4
Totals

50.5
56.1
74.5

54.8

 son that if the new hybrid (Pioneer 3541) was grown on soils amended with
 sludge at agronomic N rates, concentrations of Cd in plant tissues would
 present little, if any, impact on human food chains.  Results from this acci-
 dental choice of a hybrid with a low capacity to take up or translocate Cd to
 above ground parts reinforces previous conclusions that plant hybrids and
 cultivars can be selected and/or developed that would eliminate the potential
 health hazard associated with the uptake of Cd from sewage sludge-amended
 soils (Hinesly et al., 1981).

    Total amounts of transition and heavy metals applied on the surface of
 Blount as constituents of sewage sludge could not be accounted for within
 soil depths of 76 cm.  Further work is needed to determine how about 40% of
 the sludge-borne metals were lost.


 RESPONSES OF WINTER WHEAT AND SOYBEANS ON BLOUNT PREVIOUSLY AMENDED WITH
 ANNUAL APPLICATIONS OF SEWAGE SLUDGE

 Introduction

    Other researchers have speculated that the uptake of transition and
 heavy metals by crop plants may increase with time, after sewage sludges are
 admixed with soils, as a result of their release when organic matter decom-
 poses (Chaney, 1973; Sims and Boswell, 1978).  It was assumed that with time
 the release of sludge-borne metals would create phytotoxic conditions and/or
 increase levels of metals in plant produce  that would adversely affect animal
 and human health.

    The  purpose of  this study was  to measure the effects of sludge-borne
 metals  on  the  growth and quality  of wheat  (Triticum aestivium L.) and soy-
 beans  (Glycine max L.) where sludge applications had been  terminated.
                                         51

-------
TABLE HO.  AVERAGE CONTENTS OF MACROELEMENTS AND MINOR ELEMENTS IN CORN LEAF,
          STOVER, AND GRAIN TISSUE SAMPLES FROM BLOUNT SILT LOAM PLOTS.
 Constit-
 uent     Year
                                     Sludge Treatment
                      1/4
                      Max
                          1/2
                          Max
                           Max
              LSD
 N
1978
1979
1980
          1980
3.42
3.04
2.74
         2.87
                                     Pioneer  3517  Leaf
3.12          3.21         3.42
3.04          3.34         3.07
2.79          2.89         2.94

      Pioneer 3541 Leaf
            2.89
              2.83
3.08
              0.22*
              n.s.
              0.16**
0.19**
          1980
         1.76
                                     Pioneer  3517  Stover
1978
1979
1980

0.60
1.03
1.56

0.68
1.08
1.56

0.85
1.10
1.58
Pioneer 3541 Stover
0.92
1.18
1.83

n.s.
n.s.
n.s.

            1.73
              1.92
1.78
                                                                       n.s.
          1980
         1.81
                                     Pioneer  3517  Grain
1978
1979
1980

1.77
1.64
1.46

1.76
1.61
2.01

1.80
1.48
1.94
Pioneer 3541 Grain
1.77
1.78
1.84

n.s.
n.s.
0.37*

            1.34
              1.88
1.84
                                                                       n.s.
                                  (continued)


                                     52

-------
TABLE 10.  (continued)

Constit-
uent Year

P 1978
1979
1980



0

0.355
0.386
0.321


1/4
Max

0.326
0.362
0.304

Sludge Treatment
1/2
Max
w
Pioneer 3517 Leaf
0.352
0.404
0.311
Pioneer 3541 Leaf


Max


0.378
0.390
0.363



LSD


n.s.
n.s.
0.017**

         1980
0.280
0.257
0.276
0.319
0.014**
         1980
0.274
                                    Pioneer 3517 Stover
1978
1979
1980

0.083
0.156
0.102

0.084
0.157
0.102

0.124
0.166
0.096
Pioneer 3541 Stover
0.174
0.231
0.107

0.052**
n.s.
n.s.

0.280
0.297
0.324
                                                                       n.s.
         1980
0.362
                                    Pioneer 3517 Grain
1978
1979
1980

0.317
0.356
0.376

0.289
0.350
0.380

0.309
0.370
0.370
Pioneer 3541 Grain
0.334
0.347
0.374

n.s.
n.s.
n.s.

0.328
0.332
0.323
0.025*
                                 (continued)



                                     53

-------
TABLE 10.    (continued)
Constit-
uent Year
Ca 1978
1979
1980
1980
1978
1979
1980
1980
1978
1979
1980

0


0.884
0.734
0.526

0.542

0.347
0.443
0.403

0.295

28
21
41


1/4
Max


0.756
0.696
0.563

0.547

0.353
0.390
0.420

0.273

30
31
41

Sludge Treatment
1/2
Max
-, . .^ _______ ....
Pioneer 3517 Leaf
0.657
0.746
0.581
Pioneer 3541 Leaf
0.560
Pioneer 3517 Stover
0.301
0.456
0.344
Pioneer 3541 Stover
0.321
	 mg/kg 	
Pioneer 3517 Grain
30
21
32
_______ nig/ Kg— — — — — —
Pioneer 3541 Grain

Max


0.721
0.729
0.562

0.581

0.358
0.549
0.426

0.375

31
25
44


LSD


0.119*
n.s.
n.s.

n.s.

0.048*
0.115**
n.s.

n.s .

n.s.
n.s.
n.s.

        1980
41
33
33
33
                                                                     n.s.
                                 (continued)





                                     54

-------
TABLE 10.    (continued)


Constit-
uent.

tfc





Year

1978
1979
1980



0

0.241
0.223
0.265


1/4
Max


0.242
0.214
0.288

Sludge Treatment
1/2
Max Max LSD
m
Pioneer 3517 Leaf
0.219 0.209 n.s.
0.210 0.210 n.s.
0.263 0.263 n.s.
Pioneer 3541 Leaf
         1980     0.308       0.320         0.299        0.278         0.027*
                                    Pioneer 3517 Stover
1978
1979
1980

0.
0.
0.

171
205
231

0.
0.
0.

206
221
254

0
0
0
Pioneer
.179
.203
.202
3541
0.
0.
0.
Stover
187
216
220

n.
n.
n.

s.
s.
s.

         1980     0.157       0.170         0.187        0.178         n.s.
                                    Pioneer  3517 Grain
1978
1979
1980

0.
0.
0.

127
145
155

0.
0.
0.

119
133
158

0.121
0.138
0.148
Pioneer 3541 Grain
0.
0.
0.

127
126
149

n
n
n

.s.
.s.
.s.

         1980     0.141        0.131         0.128         0.124          0.011*
                                 (continued)




                                     55

-------
TABLE 10.  (continued)

Constit-
uent. Year


Fe 1978
1979
1980

1980

1978
1979
1980

1980

1978
1979
1980



0



120
118
124

128

105
128
129

120

16
22
25


1/4
Max



103
111
119

120

114
121
147

112

20
25
27

Sludge Treatment
1/2
Max

_—.-.-._-.— jjjgy Kg _-.______,
Pioneer 3517 Leaf
126
115
118
Pioneer 3541 Leaf
116
Pioneer 3517 Stover
129
130
100
Pioneer 3541 Stover
172
Pioneer 3517 Grain
20
32
25
Pioneer 3541 Grain
.

Max LSD



106 n.s.
115 n.s.
117 n.s.

124 8*

103 n.s.
135 n.s.
104 n.s.

113 n.s.

24 5**
25 n.s.
32 5*

        1980
                    30
31
30
31
                                                                     n.s.
                                 (continued)





                                     56

-------
TABLE 10.   (continued)
Sludge Treatment
Constit-
uent Year


Mn 1978
1979
1980

1980

1978
1979
1930

1980

1978
1979
1980


0



75
69
103

100

59
70
55

58

5.4
9.6
9.0

1/4
Max


Pioneer
50
35
44
Pioneer
52
Pioneer
48
48
32
Pioneer
32
Pioneer
4.4
6.6
6.8
Pioneer
1/2
Max

g/ Kg-— 	
3517 Leaf
46
38
56
3541 Leaf
56
3517 Stover
42
52
44
3541 Stover
37
3517 Grain
4.0
6.4
5.9
3541 Grain

Max LSD



60 n.s.
44 19*
63 n.s.

65 32*

56 n.s.
67 n.s.
48 12*

46 16*

4.0 1.0**
5.7 2.2**
7.4 2.0*

         1980
7.8
6.1
5.6
5.9
0.8**
                                  (continued)




                                      57

-------
TABLE 10-   (continued)


Constit-
uent Year



Zn 1978
1979
1980

1980

1978
1979
1980

1930

1978
1979
1980




0



73
59
57

44

39
26
52

62

30
26
28



1/4
Max



139
130
125

98

127
146
156

172

39
42
42


Sludge Treatment
1/2
Max


Pioneer 3517 Leaf
216
247
209
Pioneer 3541 Leaf
190
Pioneer 3517 Stover
248
234
370
Pioneer 3541 Stover
312
Pioneer 3517 Grain
50
49
48
Pioneer 3541 Grain



Max



308
330
289

288

350
348
393

396

58
49
58




LSD



63**
61**
50**

50**

54**
93**
162**

88s**

6**
9**
8**

         1980
33
41
48
52
8**
                                 (Continued)





                                     58

-------
TABLE 10.    (-continued)
Sludge Treatment
Constit-
uent Year 0



Cd 1978 O.S
1979 0.4
1980 0.4

1/4
Max



9.0
7.7
5.6

1/2
Max
	 mg/ kg 	
Pioneer 3517 Leaf
L7.3
21.2
14.7
Pioneer 3541 Leaf

Max LSD



20.0 6.8**
33.8 11.5**
22.7 3.8**

         1980
 0.208
 0.958
3.45
 7.72
1.10**
         1978
         1979
         1980
         1980
 0.484
                                    Pioneer 3517 Stover
1.1
0.9
0.7

6.7
10.5
7.5

13.3
25.9
26.6
Pioneer 3541 Stover
27.8
42.7
36.9

6.3**
11.3**
11.5**

 3.76
8761
1471
2746**
         1978
         1979
         1980
         1980
<0.062
                                    Pioneer 3517 Grain
0.09
<0.06
<0.06

0.18
0.25
0.32
Pioneer
0.46
0.43
0.57
3541 Grain
0.68
0.72
0.73

0.20**
0 . 20**
0.19**

<0.062
0.111
 0.171
0.086**
                                  (continued)


                                     59

-------
TABLE 10.  (continued)


Constit-
uent Year


Cu 1978
1979
1980

1980

1978
1979
1980

1980

1978
1979
1980



0



10.4
9.4
7.7

8.37

2.9
6.0
4.5

5.40

2.0
2.2
1.8


1/4
Max



11.5
11.0
9.8

10.5

3.8
7.2
6.4

7.26

2.2
2.3
1.8

Sludge Treatment
1/2
Max
— —————— mtr /Vcr— — —
—————— mgy K.g~—
Pioneer 3517 Leaf
14.0
11.6
10. 1
Pioneer 3541 Leaf
11.2
Pioneer 3517 Stover
5.0
7.3
6.6
Pioneer 3541 Stover
6.95
Pioneer 3517 Grain
2.1
2.4
1.5
Pioneer 3541 Grain


Max



17.3
12.5
12.9

13.2

5.1
8.9
6.8

7.19

1.7
2.0
1.5



LSD



2.6**
1.6**
1.9**

0.91**

1.2**
n.s.
1.6*

0.94**

0.3*
n.s.
n.s.

          1980
1.58
1.69
1.56
1.52
                                                                       n.s.
                                 (continued)
                                     60

-------
TABLE 10.   (continued)
                                   Sludge Treatment
Constit-
uent Year 0
1/4 1/2
Max Max Max
LSD

Pioneer 3517 Leaf
Ni 1978 <0.6
1979 <0.6
1980 <0.6
1980 <0.6
1978 0.9
1979 <0.6
1980 <0.6
1980 <0.6
1978 <0.6
1979 <0.6
1980 <0.6
1980 <0.6
<0.6 0.9 1.2
<0.6 <0.6 1.3
<0.6 <0.6 0.9
Pioneer 3541 Leaf
<0.6 <0.6 0.7
Pioneer 3517 Stover
0.8 2.2 2.4
<0.6 <0.6 1.7
<0.6 1.0 2.4
Pioneer 3541 Stover
<0.6 <0.6 1.6
Pioneer 3517 Grain
0.7 0.7 2.5
<0.6 1.3 2.1
0.7 1.0 2.8
Pioneer 3541 Grain
<0.6 0.7 2.0
(continued)
61
0.5**
0.4**
0.4*
n.s.
n.s.
0.4**
0.8**
0.8**
1.1**
0.4**
0.6**
0.8**

-------
TABLE 10.    (continued)

Sludge Treatment
Constit-
uent Year


Cr 1978
1979
1980

1980

1978
1979
1980

1980

1978
1979
1980

0



0.2
0.9
0.4

<0.12

0.8
0.5
0.2

0.15

0.30
<0.12
<0.12

1/4
Max


Pioneer
0.6
0.9
0.3
Pioneer
<0.12
Pioneer
0.7
0.6
0.3
Pioneer
<0.12
Pioneer
0.30
<0.12
<0.12
Pioneer
1/2
Max
lg/kg 	
3517 Leaf
0.3
1.0
0.5
3541 Leaf
0.26
3517 Stover
1.0
0.8
0.4
3541 Stover
<0.12
3517 Grain
<0.13
0.13
<0.12
3541 Grain
Max



0.2
1.0
0.6

0.38

1.1
0.9
0.2

0.17

0.16
0.17
0.16

LSD



n.s.
n.s.
n.s.

0.23**

n.s.
n.s.
n.s.

n.s.

n.s.
n.s.
n.s.

        1980
<0.12
0.25
<0.12
0.34
                                                                      n.s.
                                  (continued)




                                     62

-------
TABLE 10.   (continued)

Sludge Treatment
Constit-
uent. Year


Pb 1978
1979
1980

1980

1978
1979
1980

1980

1978
1979
1980

1980
0



1.4
0.8
0.7

<0.62

1.5
2.2
2.7

1.52

<0.6
<0.6
<0.6

<0.62
1/4
Max


Pioneer
1.8
1.0
0.7
Pioneer
<0.62
Pioneer
1.6
1.9
1.8
Pioneer
1.26
Pioneer
<0.6
1.0
<0.9
Pioneer
<0.62
1/2
Max
ig/kg 	
3517 Leaf
1.6
1.4
0.6
3541 Leaf
<0.62
3517 Stover
1.4
2.4
2.4
3541 Stover
1.69
3517 Grain
<0.6
1.3
<0.6
3541 Grain
0.70
Max



1.6
0.7
0.7

0.66

1.2
2.6
1.7

1.30

0.7
2.0
<0.6

<0.62
LSD



n.s.
n.s.
n.s.

n.s.

n.s.
n.s.
n.s.

n.s.

n.s.
n.s.
n.s.

n.s.

n.s. = no significant differences.
*, ** = significant different at P_0.05 and P_0.01, respectively.
                                      63

-------
Methods and Materials

   Details of this study were presented in a previous report (Hinesly and
Hansen, 1981).  The study was established in 1969 on a phosphorus deficient
Blount silc loam soil as a split plot completely randomized block design with
three replications of five treatments.  One-half of the split plots received
a broadcast application of 118 kg P/ha each year and the treatments on both
plot halves were:  1) irrigation with maximum amounts of digested sewage
sludge; 2) one-half maximum amounts of sludge; 3) one-fourth maximum amounts
of sludge; 4) irrigation with well water in amounts equivalent to maximum
sludge irrigation; and 5) no irrigation.  Soybeans were planted each year on
these plots from 1969 through 1976.  In 1972, soybeans planted on plots
treated with maximum amounts of digested sludge and broadcast applications of
superphosphate suffered a severe F-toxicity.  Thereafter sludge applications
were terminated on split plots not previously treated with superphosphate,
but were continued as before on the other half of each plot until the end of
1976.  After soybeans were harvested in 1976, winter wheat was seeded each
fall, but soybeans were planted again in 1978 after winter freezing and thaw-
ing had severely reduced wheat stands.

   When sludge applications were terminated in 1972 on one-half of split
plots and in 1976 on the other half, accumulative applications of sludge
solids (dry weight basis) on maximum-treated plots amounted to 242 and 411
mt/ha, respectively.


Results and Discussion

   An examination of data presented in Table 11 shows that previous sludge
applications had increased concentrations of organic-C,  N, and Ni in 0 to 15
cm depths of Blount and concentrations of P, Cd, Cu,  Cr,  Pb and Zn in 0 to 30
cm depths.  In a comparison with those previously reported for 1977 (Hinesly
and Hansen, 1981), these data showed small but significant decreases in con-
centrations of organic-C, N, P, Cd, Cr,  Pb, and Zn in the 0 to 15 cm depth by
maximum-sludge-treated Blount, but concentrations at deeper depths remained
unchanged with time.  Amounts of transition and heavy metals removed in soy-
beans and wheat can not account for losses nor is there any indication that
they migrated to lower soil depths.

   Soil pH was decreased by the two higher sludge loading rates,  but it is
unlikely Chat differences markedly affected the uptake of trace elements by
soybeans and wheat.

   Previous sludge applications had no effect on soybean and wheat grain
yields (Table 12).  Yields were about the same, regardless of when sludge
applications had been terminated.  But wheat stover yields were increased
during the last year of the study where sludge was previously applied.

   Of the several chemical element concentrations determined in soybean
tissue (Table 13), those for Cd, Cu, Ni, and Zn were affected to  the greatest
extent by previous sludge applications.   Concentrations  of Cd, Ni, and Zn in
soybean tissues from plots where sludge applications were terminated last had

                                       64

-------
    TABLE  11. TOTAL CONTENTS  OF  SELECTED  ELEMENTS  AND  pH DETERMINED IN 0 TO 15,  15  TO 30.  30 TO 46,  AND  61
               TO  76 cm SOIL DEPTH SAMPLES FROM BLOUNT  SILT LOAM PLOTS  PLANTED TO SOYBEANS  AND WHEAT.a/b/
ON
in

Sludge Applications (1969-76)
Para-
meter
Depth
cm
Organlc-
C 0-15



15-30
30-46
61-76

Year HZO

1978 0.93
1979 0.93
I960 0.93
1978 0.66
1979 0.76
1980 0.65
1978 0.38
1979 0.37
1978 0.38
1979 0.36

0

1.17
1.18
1.15
1.15
1.10
1.02
0.74
0.68
0.32
0.30

1/4
Max

1.34
1.42
1.37
1.13
1.06
1.03
0.47
0.44
0.40
0.33

1/2
Max

1.80
1.77
1.69
1.19
1.25
1.16
0.50
0.51
0.39
0.33
Sludge
Max

2.12
2.29
2.09
1.27
1.23
1.09
0.48
0.39
0.36
0.34
Sludge Applications (1969-72)
and Water Treatments
LSD .

0.52**
0.58**
0.62**
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
u2o

0.90
0.90
0.84
0.61
0.58
0.56
0.48
0.45
0.33
0.39
0

1.27
1.28
1.20
1.26
1.10
1.20
1.39
1.31
0.43
0.41
1/4
Max

1.25
1.32
1.25
1.09
1.14
1.14
0.81
0.90
0.44
0.41
1/2
Max

1.42
1.44
1.37
1.02
1.09
1.00
0.78
0.73
0.36
0.34
Max

1.82
1.74
1.60
0.86
0.94
1.02
0.38
0.38
0.38
0.33
LSD

0.59**
0.39*
0.35*
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
                                                     (continued)

-------
TABLE 11.    (continued)
Sludge Applications (1969-76)
Sludge
Para- Depth Year H_0
meter cm
N 0-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979

0.101
0.077
0.092
0.080
0.073
0.078
0.060
0.057
0.046
0.050
0

0.134
0.117
0.110
0.101
0.110
0.091
0.087
0.080
0.058
0.053
1/4
Max

0.136
0.137
0.124
0.122
0.110
0.100
0.068
0.063
0.059
0.050
1/2
Max

0.158
0.160
0.152
0.130
0.120
0.107
0.069
0.067
0.063
0.057
Max

0.231
0.217
0.189
0.137
0.127
0.108
0.067
0.057
0.057
0.050
Sludge Applications (1969-72)
and Water Treatments
LSD

0.075**
0.069**
0.047**
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
HO 0 1/4 1/2 Max LSD
Max Max
„
0.098
0.083
0.092
0.079
0.077
0.069
0.068
0.063
0.048
0.053

0.122
0.123
0.105
0.127
0.123
0.113
0.143
0.110
0.072
0.063

0.126
0.127
0.115
0.111
0.110
0.107
0.093
0.097
0.066
0.063

0.137
0.137
0.128
0.134
0.110
0.101
0.096
0.080
0.062
0.050

0.184
0.163
0.148
0.102
0.103
0.111
0.059
0.060
0.057
0.043

0.051**
0.042**
0.029*
n.s.
n.s.
n.s.
n.s.
n.s.
0.014*
n.s.
                                                 (continued)

-------
TABLE 11.    (continued)

Sludge Applications (1969-76)


Para- Depth Year
meter cm

H20

0

1/4
Max

1/2
Max
Sludge
Max
Sludge Applications (1969-72)
and Water Treatments
LSD .
H20
0
1/4
Max
1/2
Max
Max
LSD



P 0-15 ,
,

I
15-30


30-46

61-76


1978
1979
1980

1978
1979
1980
1978
1979
1978
1979

505
548
499

361
426
436
348
364
353
338

574
600
546

398
506
453
323
398
344
406

1016
1180
1080

749
765
685
369
388
377
322

1651
1780
1910

946
1070
922
329
395
373
362

3178
2970
2560

1227
1250
1030
369
376
367
338

430**
359**
368**

535*
564**
217**
n.s.
n.s.
n.s.
n.s.

491
453
503

464
416
441
367
451
322
336

493
538
559

558
527
484
508
571
463
412

626
818
981

518
658
635
440
479
370
382

983
1210
1260

633
759
633
406
436
323
392

1908
1930
1790

955
977
903
353
409
343
309

404*
192**
499**

152*
223**
266**
n.s.
n.s.
n.s.
n.s.
                                                  (continued)

-------
     TABLE 11.   (continued)
09

Sludge Applications (1969-76)
Sludge Applications (1969-72)
Sludge and Water Treatments
Para- Depth Year
meter cm
K 0-15 1978
1979
1980
15-30 1978
1979
.1980
30-46 1978
1979
61-76 1978
i979
HO 0 1/4
Max

2.00
2.04
1.89
2.07
2.20
2.06
•2.40
2.18
2.. 5 7
2.86

2.02
1.88
1.89
1.99
2.11
1.90
2.14
2.15
2.22
2.27

1.99
2.01
1.86
2.04
2.06
1.95
2.23
2.27
2.31
2.49
1/2
Max

1.98
1.99
1.92
2.00
2.08
2.00
2.13
2.16
2.38
2.42
Max

1.94
1.96
1.86
2.01
2.02
1.97
2.23
2.22
2.34
2.51
LSD .

Z__
n.s.
n.s.
n.s.
n.s.
0.11*
n.s.
n.s.
n.s.
n.s.
n.s.
n2o

2.07
2.06
2.00
1.97
2.24
2.28
2.49
2.59
2.10
2.60
0

2.01
2.02
1.90
1.96
2.09
1.89
2.08
2.19
2.16
2.25
1/4
Max

2.05
2.15
1.92
2.16
2.19
1.93
2.25
2.38
2.26
2.43
1/2
Max

1.99
2.00
1.88
1.97
2.04
1.95
2.10
1.98
2.91
2.49
Max

1.97
2.03
1.90
1.94
2.19,
2.05-
2.13
2.28
2.27
2.01
LSD

n.s.
n.s.
n.s.
n.s.
n.s.
0.27**
n.s.
0.334*
n.s.
n.s.
                                                      (continued)

-------
                 TABLE 11.    (continued)
            o

Sludge Applications (1969-76)
Sludge
Para-
meter
Na



Depth Year
cm
0-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979
»2°

0.836
0.778
0.812
0.674
0.721
0.731
0.631
0.524
0.515
0.495
0

0.872
0.767
0.879
0.861
0.828
0.827
0.864
0.764
0.712
0.709
1/4
Max

0.837
0.808
0.839
0.816
0.774
0.794
0.773
0.681
0.576
0.611
1/2
Max

0.828
0.782
0.840
0.797
0.770
0.747
0.748
0.686
0.571
0.587
Max

0.791
0.752
0.797
0.756
0.717
0.758
0.702
0.628
0.562
0.485
Sludge Applications (1969-72)
and Mater Treatments
LSD . H20

n.s. 0.774
n.s. 0.706
0.047* 0.780
0.106* 0.535
0.075* 0.694
n.s. 0.608
n.s. 0.574
n.s. 0.475
n.s. 0.374
n.s. 0.416
0

0.864
0.807
0.857
0.799
0.828
0.842
0.852
0.791
0.736
0.715
1/4
Max

0.837
0.828
0.824
0.797
0.753
0.821
0.745
0.664
0.522
0.574
1/2
Max

0.841
0.787
0.830
0.816
0.758
0.805
0.771
0.628
0.832
0.650
Max

0.792
0.762
0.779
0.663
0.709
0.731
0.759
0.524
0.539
0.421
LSD

n.s.
n.s.
n.s.
n.s.
n.s.
0.139**
n.s.
n.s.
n.s.
0.207*
                                                                    (continued)
•< •"Am i-  i

-------
TABLE  11.   (continued)

Sludge Applications (1969-76)
Sludge and Water
Para- Depth Year
meter cm
Ca 0-15 (1978
1 i1"9
1 1 1980
1
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979
H20

0.449
0.366
0.489
0.406
0.414
0.422
0.391
0.420
1.52
1.22
0 1/4
Max

0.463
0.406
0.432
0.445
0.452
0.399
0.399
0.393
0.394
0.455

0.516
0.425
0.500
0.463
0.498
0.409
0.385
0.452
1.08
1.14
1/2
Max

0.544
0.470
0.489
0.491
0.507
0.417
0.420
0.347
0.490
0.598
Max

0.599
0.525
0.570
0.542
0.493
0.424
0.409
0.370
1.25
1.22
Sludge Applications (1969-72)
Treatments
LSD . U20 0

0.072**
0.098**
n.s.
n.s.
0.062*
n.s.
n.s.
n.s.
n.s.
n.s.

0.487 0.451
0.456 0.402
0.411 0.457
0.346 0.457
0.420 0.461
0.381 0.415
0.586 0.487
0.630 0.406
2.94 0.466
2.73 0.425

1/4
Max

0.517
0.470
0.478
0.466
0.493
0.433
0.443
0.455
0.959
1.06

1/2
Max

0.518
0.461
0.506
0.472
0.516
0.410
0.411
0.321
0.414
0.471

Max

0.745
0.516
0.542
0.415
0.516
0.422
0.426
0.412
1.55
2.03

LSD

0.200**
n.s.
0.040**
n.s.
n.s.
n.s.
n.s.
0.148*
n.s.
n.s.
                                                 (continued)

-------
TABLE 11.    (continued)

Sludge Applications (1969-76)
Para-
meter
Mg



Depth Year
cm
0-15 , 1978
i 1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979

H20 0

0.391 0.366
0.342 0.330
0.437 0.374
0.496 0.360
0.538 0.333
0.604 0.387
0.729 0.444
0.677 0.418
1.15 0.629
1.10 0.5:33

1/4
Max

0.384
0.359
0.404
0.424
0.370
0.467
0.565
0.528
0.957
1.00

1/2
Max

0.410
0.355
0.417
0.448
0.389
0.557
0.633
0.518
0.737
0.851
Sludge
Max

0.448
0.388
0.452
0.526
0.466
0.549
0.705
0.615
0.921
0.979
Sludge Applications (1969-72)
and Water Treatments
LSD .

%_
n.s.
n.s.
n.s.
n.s.
n.s.
0.121*
n.s.
n.s.
n.s.
n.s.
H20 0

0.495 0.369
0.433 0.334
0.511 0.389
0.464 0.343
0.604 0.344
0.778 0.379
0.938 0.461
0.814 0.382
1.52 0.582
1.31 0.563
1/4
Max

0.441
0.380
0.431
0.456
0.421
0.410
0.548
0.520
0.841
0.983
1/2
Max

0.415
0.382
0.428
0.419
0.407
0.463
0.585
0.507
0.686
0.726
Max

0.640
0.405
0.468
0.433
0.497
0.561
0.605'
0.568
1.29
1.18
LSD

n.s.
n.s.
n.s.
n.s.
0.151*
0.275**
n.s.
n.s.
n.s.
n.s.
                                                 (continued)

-------
TABLE  11.   (continued)

Sludge Applications (1969-76)
Sludge Applications (1969-72)
Sludge and Water Treatments
Para-
meter
Fe



Depth Year
cm
! :
0-15 1978
j 1979
1980
15-30 1978
1 1979
1980
30-46 1978
1979
61-76 1978
1979
V

1.91
2.07
2.18
3.01
2.83
2.96
4.18
3.94
4.09
4.26
0

1.80
1.83
1.77
1.92
1.91
1.77
2.18
2.42
3.49
3.26
1/4
Max

2.00
2.04
2.04
2.32
2.21
2.45
3.14
3.07
4.02
4.26
1/2
Max

2.31
2.13
2.03
2.38
2.21
2.15
3.12
3.41
4.53
4.18
Max

2.56
2.35
2.21
2.66
2.50
2.60
3.65
3.78
3.99
4.02
LSD .

0.50**
0.31*
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
H20

2.34
2.48
2.49
3.46
3.44
3.72
4.28
4.44
3.21
3.92
0

1.96
1.84
1.95
1.97
1.98
1.76
2.26
2.28
3.46
3.37
1/4
Max

2.20
2.30
2.10
2.62
2.52
2.37
3.15
3.07
3.70
3.84
1/2
Max

2.10
2.06
2.01
2.24
2.22
2.10
2.99
2.77
4.80
4.02
Max

2.41
2.40
2.38
2.80
2.80
3.04
2.95
3.93
3.76
3.24
LSD

n.s.
n.s.
n. s.
n.s.
0.78*
1.12*
n.s.
n.s.
n.s.
n.s.
                                                 (continued)

-------
    TABLE  11.    (continued)
                          Sludge Applications (1969-76)
                               Sludge Applications (1969-72)
                                                 Sludge and Water Treatments
    Para-   Depth  Year
    meter    cm
Max
1/2
Max
              Max
LSD .
1/4
Max
1/2
Max
Max
LSD
to

0-15


15-30


30-46

61-76

i
1978
1979
1980
1978
1979
1980
1978 '
1979
1978
1979

1180
1120
1220
944
721
865
544
422
681
640

1630
1210
1100
1330
1240
1060
935
793
447
429

1510
1310
1260
1300
1200
1160
829
621
465
473

1640
1140
1190
1410
980
1020
793
594
604
597

1360
1060
1070
984
758
802
698
672
504
483
	 mg/
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
Kg 	
1200
953
1020
783
659
669
612
602
520
617

1580
1200
1220
1740
1420
1240
1810
1390
1190
742

1430
1200
1190
1360
1380
1320
1200
1110
531
640

1200
1040
1130
1190
829
863
1060
737
518
714

1420
1040
1050
888
803
884
805
485
554
476

n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
446*
n.s.
                                                    (continued)

-------
TABLE  11.   (continued)

Sludge Applications (1969-76)
Para-
meter
Zn



Depth Year
cm
0-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979

H20

62
67
70
83
71
78
98
87
99
106

0

68
76
71
61
69
67
75
52
92
83

1/4
Max

152
156
157
135
108
111
88
71
101
108

1/2
Max

275
255
255
181
174
155
99
91
109
107
Sludge
Max

448
409
382
226
213
186
118
99
99
102
Sludee Applications (1969-72)
and Water Treatments
LSD .

59**
29**
48**
101**
58**
31**
n.s.
n.s.
n.s.
14*
H20

70
70
70
82
79
92
106
83
96
9b
0

66
77
77
83
74
73
80
64
100
89
1/4
Max

114
125
126
108
103
105
91
78
103
105

1/2
Max

189
190
187
143
132
116
97
73
103
100

Max

340
308
295
175
192
167
101
97
102
82

LSD

62**
29**
29**
27**
20**
• 23**
n.s.
15**
n.s.
n.s.
                                                 (continued)

-------
     TABLE 11.    (continued)
Ul
Sludge Applications (1969-76)
Sludge Applications (1969-72)
Sludge and Water Treatments
Para- Depth Year 1120
meter cm
0
1/4
Max
1/2
Max
Max
LSD .
H20
0 1/4 1/2 Max LSD
Max Max


Cd 0-15 1978 0.43
1979 <0.25
1980 
-------
TABLE  11.   (continued)

Sludge Applications (1969-76)

Para-
meter

Cu










Depth Year
cm
-
'o-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979

H20



18
22
18
28
20
18
31
29
34
35

0



19
21
19
19
26
12
20
18
28
27

1/4
Max


46
52
47
44
35
27
31
22
31
33

1/2
Max


87
86
81
57
51
39
29
21
35
32
Sludge
Max



148
154
129
78
67
52
29
25
29
26
and Hater
LSD .

>,
- -mg/Kg-
22**
16**
19**
29**
22**
10**
n.s.
n.s.
n.s.
n.s.
Sludge Applications (1969-72)
Treatments
H_0



18
23
21
27
25
25
36
34
33
34
0



18
22
21
20
22
14
20
18
30
23
1/4
Max


34
39
33
33
31
25
27
22
31
29
1/2
Max


56
62
54
41
35
26
26
21
31
34
Max



102
100
93
55
56
47 .
24
31
31
27
LSD



14*
11**
10**
9**
5**
7**
n.s.
11*
n.s.
n.s.
                                                  (continued)

-------
TABLE 11.    (continued)

Sludge Applications (1969-76)

Para-
meter

Depth Year H20
cm

0


1/4
Max

1/2
Max
Sludge*
Max

Sludge Applications (1969-72)
and Water Treatments
LSD

H20

0

1/4
Max
1/2
Max
Max

LSD



Ni









0-15 1978 18
1979 13
1980 28
15-30 1978 28
1979 22
1980 27
30-A6 1978 36
1979 30
61-76 1978 42
1979 40
16
13
25
23
19
23
22
20
28
20
23
19
33
28
22
28
29
26
33
28
26
24
39
30
25
29
25
24
38
30
38
36
50
34
29
32
32
28
37
34
4**
5**
4**
n.s.
6*
n.s.
n.s.
n.s.
n.s.
n.s.
18
15
31
28
30
35
47
44
41
36
19
13
27
27
22
21
30
26
29
22
17
15
31
26
24
27
33
33
37
30
21
18
34
25
21
25
28
26
38
34
24
25
42
31
28
33
29
36
40
29
n.s.
4**
6**
n.s.
n.s.
9**
n.s.
13**
U.S.
n.s.
                                                (continued)

-------
     TABLE 11.   (continued)
oo

Sludge Applications (1969-76)

Para- Depth Year H.O
meter cm

0


1/4
Max

1/2
Max
Sludge
Max

Sludge Applications (1969-72)
and Water Treatments
LSD

H2°

0

1/4
Max
1/2
Max
Max

LSD



Cr 0-15 1978 43
1979 36
1980 43
15-30 1978 49
1979 43
1980 30
30-46 1978 58
1979 56
61-761978 58
1979 62
46
38
32
43
37
23
40
36
43
51
119
95
86
76
59
40
51'
50
60
72
202
141
146
96
95
53
54
4B
49
60
354
232
261
126
117
68
56
47
55
52
31**
29**
40**
41**
30**
14**
n.s.
n.s.
n.s.
14*
47
38
45
51
49
36
63
56
70
60
44
37
57
38
37
24
39
36
50
52
71
66
72
59
56
44
50
49
65
57
116
105
114
76
73
43
48
45
52
55
199
167
195
90
100
64
57
67
68
48
29**
14**
55**
14**
14**
• 12**
12*
n.s.
n.s.
n.s.
                                                       (continued)

-------
TABLE  11.   (continued)

Sludge Applications (1969-76)

Para-
meter

Pb










Depth
cm

0-15
1

15-30


30-46

61-76


Year

|
1 1978
1979
1980
1978
1979
1980
1978
1979
1978
1 1979

H20



25
23
22
31
23
18
18
21
23
20

0



24
23
26
30
28
20
15
17
17
18

1/4
Max


42
46
46
48
36
26
17
19
18
19

1/2
Max


68
67
72
58
39
32
19
19
25
19
Sludge
Max



111
107
101
68
53
39
18
19
22
22
Sludge Applications (1969-72)
and Water Treatments
LSD .

- nig/kg
23**
8**
15**
25**
15*
10**
n.s.
n.s.
n.s.
n.s.
"2°



22
23
23
30
23
20
20
22
19
18
0



21
24
32
30
24
18
16
18
19
15
1/4
Max


35
37
39
38
38
31
17
18
18
18
1/2
Max


50
53
55
47
37
26
17
18
25
20
Max



89
82
84
50
48
39 .
16
20
20
18
LSD



26**
6**
15**
10**
12**
8**
n.s.
n.s.
n.s.
n.s.
                                                 (continued)

-------
    TABLE  11.   (continued)
g

Sludge Applications (1969-76)
Para- Depth Year
meter cm

PH 0-15 1978
1979
1980
15-30 1978
1979
1980
30-46 1978
1979
61-76 1978
1979

H20

6.3
6.7
6.7
5.2
5.3
5.6
4.8
5.2
6.1
6.4

0

6.1
6.6
6.6
5.4
5.8
5.8
4.6
5.0
4.5
4.8

1/4
Max

5.6
6.4
6.4
5.2
5.6
5.5
4.6
4.8
5.7
5.7

1/2
Max

5.5
6.1
6.1
5.0
5.4
5.3
4.4
4.8
5.2
5.5
Sludge
Max

5.1
5.9
6.0
4.6
5.0
5.1
4.3
4.5
5.3
5.7
and Water
LSD .

0.8**
0.4**
0.4*
0.4*
0.5*
0.4*
0.2*
n.s.
n.s.
n.s.
Sludge Applications (1969-72)
Treatments
H20

6.2
6.8
6.7
5.4
5.6
5.5
5.9
6.1
7.0
7.1
0

6.1
6.6
6.6
5.8
5.7
5.9
4.7
5.1
4.6
5.0
1/4
Max

6.3
6.6
6.6
5.7
5.7
5.9
5.1
5.4
5.5
5.9
1/2
Max

6.0
6.4
6.5
5.5
5.5
5.4
4.6
4.8
5.3
5.5
Max

5.8
6.3
6.4
5.2
5.3
5.5
4.7
5.0
6.3
6.5
LSD

n.s.
n.s.
n.s.
n.s.
n.s.
0.4**
0.9*
n.s.
n.s.
n.s.

     aj   Concentrations are on a dry-weight basis.
     _b/   After 1973,  P applications were terminated and sludge applications  were continued after 1972 only on
         split-plots  formerly treated with superphosphate.
           Significantly different at P<0.05 and P<0.01, respectively.
* **

-------
 TABLE  12.  SOYBEAN YIELDS  AND WHEAT GRAIN AND STOVER YIELDS  FROM PLOTS
           DESIGNATED  NW 500.
         Continued Sludge Applications     Terminated Sludge Applications
      	Sludge Application Rates	
                   1/4   1/2                          1/4   1/2
Year   H2
-------
higher concentrations than those from plots which received sludge for a
shorter time.  But Cu concentrations were not significantly different between
the two sludge application periods.  Copper concentrations in leaf and
petiole and beans were significantly less on plots treated with maximum
amounts of sludge as compared to similar tissues from control plots.
Relative to concentrations previously reported (Hinesly and Hansen,  1981),
Ni, Cd, and Zn decreased most rapidly in all- soybean tissues after sludge
applications were terminated by order of listing.  Bean-Zn concentrations did
not decrease after sludge applications were terminated.   Six years after
sludge applications were suspended, both Zn and Cd concentrations remained at
significantly higher levels in beans from sludge-treated Blount as compared
to those from control plots.

   Cadmium, Cu, Ni, and Zn were the elements whose concentrations in wheat
grain and residues were most enhanced by sludge applications.  In comparing
the concentrations of these elements in wheat tissues collected during the
last year of the study with those three years earlier,  as reported by Hinesly
and Hansen (1981), for the same variety, it appears that concentrations of Ni
and Cu receded very rapidly toward background levels after sludge applica-
tions were terminated.  This was not the case for Cd and Zn concentrations.
These two elements were readily available for uptake by wheat eight years
after sludge applications were suspended.


Summary and Conclusion

   After the second year following the suspension of sludge applications,
organic matter contents of sludge amended Blount did not change signifi-
cantly.  Concentrations of Cd, Ni,  and Zn soybean tissues decreased precipi-
tously the first year or two after sludge applications  were suspended,  but
very little after that time.  Concentrations of Cd and Zn in wheat tissues
from plots formerly treated with sludge did not decrease with time,  but Ni
and Cu concentrations did.

   Nickel concentrations were higher in the bean of soybeans and grain of
wheat than in foliage tissues.  Concentrations of Cu were decreased in soy-
beans and increased in wheat as a result of sludge applications.

   If the uptake of transition and heavy metals by soybeans and wheat were
a problem, there is no indication that the situation worsened after termina-
tion of sludge applications.

   Further work is needed to see if Cd uptake by wheat  could be reduced by
liming the sludge-amended Blount soil.


RESPONSES OF CONTINUOUS CORN ON STRIP-MINE SPOIL WITH AND WITHOUT ANNUAL
APPLICATIONS OF DIGESTED SEWAGE SLUDGE

Intraduetion

   Strip-mined lands have low contents of organic matter.  Nitrogen levels
are too low in freshly graded spoil to support nonleguminous plants.   In

                                       82

-------
     TABLE 13.  CONCENTRATIONS OF SELECTED CHEMICAL ELEMENTS OF LEAF AND PETIOLE, BEAN, AND STALK SAMPLES
                FROM SOYBEANS (BEESON CULTIVAR) AND LEAF, GRAIN, AND STOVER SAMPLES FROM WHEAT (ABE VARIETY)
                GROWN ON BLOUNT SILT LOAM SOIL, WITH AND WITHOUT SLUDGE.a/
oo
Constit-
uent Year

Sludge Applications (1969-76)
Sludge
HO 0 1/4 1/2 Max
Max Max

Sludge Applications (1969-72)
and Water Treatments
LSD H.O 0 1/4 1/2 Max LSD
Max Max
V
Soybean leaf and petiole
N 1978
1978
1978
1980
3.39 3.58 3.73 3.02 2.72 0.54** 2.87 3.60 3.23 3.00 2.72 n.s.
Soybean bean
6.44 6.59 6.46 6.78 6.45 n.s. 6.56 6.25 6.58 6.49 6.48 n.s.
Soybean plant residue
0.76 0.88 0.82 . 0.75 0.60 n
2.17 2.41 3.34 3.55 3.97 0
.s. 0.99 0.75 0.84 0.83 0.69 n.s.
Wheat leaf
.54** 2.32 2.59 2.45 2.64 3.13 0.33**
Wheat grain
                1979  1.89    2.03   2.47   2.52   2.81   0.51**   1.71   2.12    1.95    2.20    2.30    n.s.
                1980  1.70    1.84   2.33   2.42   2.60   0.41**   1.72   2.02    1.76    1.86    2.13    0.26*

                                                          Wheat stover

                1979  0.225   0.210  0.333  0.427  0.604   0.239**  0.226  0.210   0.222   0.263   0.315   0.043**
                1980  0.254   0.234  0.278  0.402  0.524   0.175**  0.235  0.255   0.215   0.239   0.243   n.s.

                                                    (continued)

-------
TABLE 13.   (continued)
                    Sludge Applications (1969-76) _       Sludge  Applications  (1969-72)
 Cons'tit-         _ Sludge and Water Treatments
 uent      Year   H^O      0     174    172    Max     LSD     H^O    0     1/4    172    Max     LSD
                                 Max    Max                                  Max    Max
                                                Soybean leaf and  petiole

           1978  0.194    0.232   0.191   0.095   0.136   0.069*   0.129  0.219  0.192  0.117  0.126   0.076*

                                                      Soybean bean

           1978  0.512    0.553   0.488   0.408   0.514   n.s.     0.467  0.507  0.513  0.505  0.499   n.s.

                                                  Soybean  plant residue

           1978  0.062    0.067   0.053  . 0.037   0.056   n.s.     0.044  0.056  0.048  0.044  0.038   n.s.

                                                       Wheat leaf

           1980  0.363    0.362   0.470   0.576   0.599   0.052** 0.279  0.266  0.395  0.434  0.562   0.200**

                                                       Wheat grain

           1979  0.399    0.417   0.480   0.489   0.491   0.057** 0.359  0.436  0.457  0.471  0.482   0.072*
           1980  0.438    0.443   0.485   0.472   0.484   n.s.     0.462  0.459  0.448  0.447  0.440   n.s.

                                                      Wheat  stover

           1979  0.020    0.036   0.081   0.078   0.141   0.069*   0.020  0.032  0.044  0.051  0.062   0.020**
           1980  0.049    0.065   0.080   0.132   0.182   0.064** 0.044  0.046  0.062  0.071  0.080   0.022**


                                               (continued)

-------
TABLE  13.  (continued)
                   Sludge Applicationa (1969-76) _       Sludge Applications  (1969-72)
Constit-         _ ; _ Sludge and Mater Treatments    _
uent      Year   Ho                     2    Max     LSD     HjO     0        4     12    Max     LSD
                                Max    Max                                 Max     Max

                 _________________________________________ 
-------
TABLE  13.   (continued)
                    Sludge  Applications  (1969-76)	       Sludge Applications (1969-72)	
Conatit-	Sludge and Mater Treatments	
uent      Year   H^O       6      T/4     f/2    Max     LSD     H^O     0     iff    T/2    Max     LSD
                                 Max     Max                                  Max    Max

                __________________________________________
-------
     TABLE 13.    (continued)
00

Sludge Applications
Constit-
uent Year H-0 0

(1969-76)
Sludge Applications (1969-72)
Sludge and Water Treatments
1/4
Max

1/2
Hax

Max LSD H20


0

1/4
Max

1/2
Max

Max

LSD

Soybean leaf and petiole
Fe 1978 114 122
1978 148 74
1978 255 205
1980 146 118
1979 52.0 57.2
1980 46.6 56.5
1979 85.4 95.9
1980 120 107
102
73
246 .
108
64.9
56.3
90.6
106
93
63
267
155
59.7
56.8
159
93.0
88
60
232
136
62
72
135
96
19* 90
Soybean bean
110
n.s. 148 79
Soybean plant residue
n.s. 146
Wheat leaf
n.s. 179
Wheat grain
.3 n.s. 44.3
.8 n.s. 44.4
Wheat stover
n.s. 120
.4 n.s. 165
229
118
59.7
48.4
88.1
97.7
110
79
214
114
59.7
49.8
109
95.1
94
76
222
165
62.3
42.3
101
93.6
90
82
233
126
57.1
49.4
122
99.2
14**
44*
n.s.
n.s.
9.95*
n.s.
n.s.
51.3**
                                                      (continued)

-------
   TABLE  13.   (continued)
o>
CD

Sludge Applications
Constit-
uent Year HO 0

(1969-76)
Sludge
Applications (1969-72)
Sludge and Water Treatments
1/4
Max

1/2
Max

Max LSD H20


0

1/4
Max

1/2
Max

Max LSD

— — — — ————— — — — — — — uig/ ng —
Soybean leaf and petiole
Mn 1978 84 109
1978 34 31
1978 35 44
1980 127 116
1979 28.2 29.8
1980 46.8 46.8
1979 64.3 65.6
1980 76.6 72.3
100
26
35
105
29.4
51.8
57.2
55.3
108
24
43
113
31.7
42.5
68.4
52.5
212
29
48
100
23
31
60
34
82* 59
Soybean bean
110
n.s. 24 28
Soybean plant residue
n.s. 24
Wheat leaf
n.s. 90.4
Wheat grain
.7 n.s. 24.7
.9 13.4** 39.7
Wheat stover
.0 n.s. 52.6
.8 26.2** 60.3
46
92
33
48
74
67
86
26
36
.7 115
.9 31.4
.2 51.8
.5 69.3
.3 68.1
83
24
34
101
29.4
46.1
54.4
61.0
84 n.s.
26 n.s.
34 12*
87.0 n.s.
23.4 n.s.
36.9 n.s.
50.2 n.s.
36.2 20.4*
                                                    (continued)

-------
    TABLE 13.    (continued)
                        Sludge Applicationa (1969-76)	      Sludge Applications (1969-72)	
     Constit-         	Sludge and Water Treatments	
     uent      Year  • lD>      0     T?4    T/2    Max     LSD     O0     174\J2    Max     LSD~
                                     Max    Max                                  Max    Max
                      	rag/kg	
                                                    Soybean leaf and petiole

     Zn         1978   30      41     113    168    262    34**     28     40     65     106    129     30**

                                                          Soybean bean

               1978   52      57      77     82     93    16**     49     56     62      74    76       9**

0,                                                     Soybean plant residue
VO

               1978   12      12   '   31  .   66    117    33**     11     15     19      33     40     14**

                                                           I/heat leaf

               1980   9.11   10.3    28.8   74.9  108   35.8**     7.88   9.05  15.4    32.6   75.5  12.5**

                                                           Wheat grain

               1979  36.5    44.5    87.8   99.2  113   27.1**    31.5   51.8   60.5    85.1   94.9  18.1**
               1980  34.8    38.7    79.7   91.4   99.7 25.3**    34.6   43.3   49.5    66.8   90.6  19.4**

                                                          Wheat atover

               1979   6.77   16.2   111    217    261    121**    12.6   22.1   50.3   141    206    93.4**
               1980  13.0    13.5    62.5  155    196   51.9**    12.1   14.6   31.4    74.7  152    59.5**


                                                    (continued)

-------
     TABLE 13.   (continued)
                        Sludge Applications  (1969-76)	        Sludge Applications  (1969-72)	
     Const it-         	Sludge and Water Treatments	
     uent      Year   H^O      0     174     U2    Max     LSD     fi^O     0      \ft    172    Max     LSD
                                     Max     Max                                  Max    Max

                      	Dig/kg	

                                                    Soybean leaf and petiole

     Cd        1978  0.07    0.17   1.50   2.52   8.31  0.94**   <0.06   0.17   0.57    1.63   2.45  0.51**

                                                          Soybean bean

               1978  0.13    0.18   0.39   0.60   1.30  0.32**    0.09   0.13   0.29    0.50   0.49  0.22**
vo
0                                                     Soybean plant residue

               1978  0.11    0.22   0.85  . 1.78   3.96  1.26**    0.09   0.20   0.55    1.28   1.40  0.40**

                                                           Ir/heat leaf

               1980  0.072   0.143  1.10   4.08   7.54  1.07**   <0.06   0.121  0.600   1.56   3.90  1.83**

                                                           Wheat grain

               1979  0.117   0.185  1.35   2.41   3.10  0.354**   0.100  0.176  0.635   1.64   2.66  0.901**
               1980  0.075   0.122  1.31   2.93   4.29  0.721**  <0.06   0.092  0.567   1.31   3.46  1.60**

                                                          Wheat stover

               1979  0.152   0.262  1.83   5.23   8.81  3.76**    0.165  0.335  0.952   3.10   5.22  2.00**
               1980  0.204   0.231  1.94   6.22   9.62  2.76**    0.222  0.237  0.761   2.38   7.11  3.29**


                                                    (continued)

-------
TABLE  13.   (continued)
                    Sludge Applicationa (1969-76)	       Sludge Applications (1969-72)	
 Cona'tit-	Sludge and Water Treatments    	
 uent      Year   1G>      0     174172    Max     LSD  '   O     0     l74T/2    Max     LSD
                                 Max    Max                                  Max    Max

                  	ing/kg--	

                                                Soybean leaf and petiole

 Cu         1978   5.2     6.5    6.0    4.2    4.2   0.8**     5.4    6.6    5.9    5.1    4.4    0.8**

                                                      Soybean bean

           1978  12.6    14.0   14.1   11.1   12.2   n.s.     16.7   15.1   13.5   12.6   12.4    2.6*

                                                  Soybean plant residue

           1978   2.6     2.7    2.5  .  2.6    3.1   n.s.      3.4    3.1    2.6    2.6    3.0    n.s.

                                                       Wheat leaf

           1980   5.73    5.24   6.93   8.69   8.77  2.45*     4.91   4.40   5.66   5.74   7.76   1.90*

                                                       Wheat grain

           1979   4.56    3.98   5.02   4.96   5.76  0.808*    4.04   4.38   4.67   4.84   5.47   n.s.
           1980   3.55    3.48   4.41   4.26   4.72  0.654**   3.69   3.91   3.82   3.98   4.30   n.s.

                                                      Wheat stover

           1979   2.84    2.20   2.43   2.60   5.12  n.s.      1.90   1.73   2.02   2.55   3.13   0.714*
           1980   1.35    1.28   2.18   2.45   3.48  1.13**    1.24   1.38   1.43   1.43   1.92   n.s.


                                                (continued)

-------
TABLE  13.   (continued)
                    Sludge Applications (1969-76)	       Sludge Applications (1969-72)	
Constit-         	Sludge and Water Treatments	
uent       Year  O      0     174172    Max     LSD     O     0     T74    T72    Max     LSD"
                                 Max    Max                                  Max    Max
                          -   _     _                    mg/kg

                                                Soybean leaf and petiole

Nl         1978  0.7     0.6    1.2    1.1    2.4    1.0**    0.6    1.0    1.0    1.0    0.8     n.s.

                                                      Soybean bean

           1978  4.4     5.3    6.2    8.9   16.2    4.2**    3.9    5.8    5.2    5.6    6.4     n.s.

                                                  Soybean plant residue
                                                  	f -     • i •••••• i    .T^I

           1978 <0.6    <0.6    0.8   . 1.3    3.4    1.3**   <0.6    0.7    0.6    0.9    0.8     0.4*

                                                       Wheat leaf

           1980 <0.62   <0.62  <0.62   0.698 <0.62   n.s.    <0.62  <0.62  <0.62  <0.62  <0.62    n.s.

                                                       Wheat grain

           1979  0.689   0.661  1.13   1.77   3.32   1.13**  <0.62   0.709  0.832  1.72   1.54    0.862*
           1980 <0.62   <0.62  <0.62  <0.62   1.84   0.629** <0.62  <0.62  <0.62  <0.62  <0.62    n.s.

                                                      Wheat stover

           1979  0.859   1.30   0.661  1.10   2.51   1.14*   <0.62   0.803  0.724  1.39   1.17    n.s.
           1980 <0.62    0.878 <0.62  <0.62   1.14   n.s.    <0.62  <0.62  <0.62  <0.62  <0.62    n.s.


                                                (continued)

-------
TABLE  13.   (continued)
                    Sludge Applications (1969-76)	      Sludge Applications (1969-72)	
 ConsLit-        	Sludge  and Water Treatments
 uent      Year   iTo      0      174T/2     Max     LSD     O     0      iff     T?2    MaiLSD~
                                 Max     Max                                   Max     Max
                  	mg/kg	

                                                Soybean  leaf  and  petiole

 Cr         1978   0.28    0.14    0.37    0.14    0.30    n.s.     0.28   0.40   0.54    0.71    0.61     n.s.

                                                      Soybean bean

           1978   1.46    0.75    0.82    1.04    1.39    n.s.     1.36   0.94   1.24    2.15    1.90     n.s.

                                                  Soybean  plant residue

           1978   1.2      1.4     1.5   .  2.1     1.5     n.s.     1.0    1.5     1.4     0.9     2.1      n.s.
                                                                                            I
                                                      Wheat  leaf

           1980   1.30    1.56    0.962   1.36    1.27    n.s.     0.777   1.15   1.57    1.25    1.84     n.s.

                                                      Wheat  grain

           1979   0.514   0.455   0.588   O.383  0.475   n.s.     0.573   0.463  0.576   0.774   0.537    n.s.
           1980  <0.125  <0.125  <0.125  <0.125  0.283   n.s.     0.235   0.133  0.178   0.524  <0.125    n.s.

                                                      Wheat stover

           1979  <0.125   0.199   0.230   0.276  0.293   n.s.     0.901  <0.125  0.358   0.290   0.191    n.s.
           1980   0.851   1.34    1.03    1.18    2.69    n.s.     0.533   0.534  0.728   0.679   1.26     n.s.


                                                (continued)

-------
TABLE  13.   (continued)

Sludge Applications (1969-76) Sludge Applications (1969-72)
Constit-
uent Year H.O 0 1/4 1/2
Max Max

Pb 1978 2.4 2.0 2.0 2.0
1978 <0.6 0.8 <0.6 <0.6
1978 1.3 1.2 1.5 . 1.6
1980 1.50 1.19 1.03 1.42
1979 <0.62 <0.62 <0.62 <0.62
1980 <0.62 <0.62 <0.62 <0.62
1979 1.40 <0.62 1.59 1.11
1980 <0.62 <0.62 <0.62 <0.62
Sludge and Water Treatments
Max LSD H.O 0 1/4 1/2 Max
Max Max
mn/k-n _
	 rag/ Kg 	 	
Soybean leaf and petiole
1.8 n.s. 2.5 1.8 2.6 2.3 2.4
Soybean bean
<0.6 n.s. 1.0 1.3 <0.6 <0.6 <0.6
Soybean plant residue
1.5 n.s. 1.0 1.3 1.5 1.6 1.8
I/heat leaf
2.23 n.s. 0.859 0.747 1.02 1.22 1.81
Wheat grain
<0.62 n.s. <0.62 <0.62 <0.62 0.805 <0.62
<0.62 n.s. <0.62 <0.62 <0.62 <0.62 <0.62
Wheat stover
1.01 n.s. 0.832 <0.62 0.721 0.795 1.05
<0.62 n.s. <0.62 <0.62 <0.62 <0.62 <0.62

LSD

n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
0.248**
n.s.

*,**  Significantly different at P<0.05 and P<0.01, respectively.

-------
calcareous strip-mine spoil F availability is low and the growth of many
plant species are restricted because they are unable to obtain sufficient
amounts of this major nutrient.  But strip-mined lands frequently contain
abundant quantities of available K.  Digested sewage sludges contain 40 to
50% organic matter, and, if applied at sufficiently high loading rates, can
supply ample amounts of N and P for plant growth.  Thus, it seemed reason-
able to expect that applications of sewage sludge would improve soil physical
properties, provide N and F for balanced plant nutrition with the K present
in strip-mine spoil, and the high pH of calcareous spoil would minimize the
uptake of transition and heavy metals contained in sewage sludge.

   A cooperative study with the Metropolitan Sanitary District of Chicago
was initiated in 1973 on freshly graded strip-mined spoil in Fulton County,
Illinois to determine the responses of corn to sewage sludge annually applied
by furrow irrigation.  The results obtained through 1977 from this study were
reported previously (Hinesly and Hansen, 1981).  Funding to the Department of
Agronomy, University of Illinois,  to support the continuation of this study
ended in 1979.  Thus, the purpose here is to report findings for the two
additional years of the study.


Materials and Methods

   The protocol for the calcareous strip-mine spoil study was the same as
that for the long-term continuous corn study initiated on acid Blount silt
loam in 1968 on the Northeast Agronomy Research Center, which was discussed
earlier in section 4 of this report.


Results and Discussion

   Concentrations of selected chemical elements in sludge are shown in
Tables 14 and 15.  At the end of the last growing season, 392 mt/ha (dry
weight equivalent) of sludge solids had been applied on maximum sludge-
treated plots of strip-mine spoil material.  A total of 248 mt/ha had been
applied prior to 1978.  Appropriately lesser amounts were applied on 1/4-
and I/2-maximum-treated plots.  Maximum sludge loading rates supplied more
than 3,000 and 2,000 kg/ha of N and F, respectively, during each of the last
two years.  During the first 5 years, 1973 through 1977, accumulative sludge-
borne Cd, Cu, Ni, and Zn applied on maximum-treated plots amounted to 87, 426,
108, and 1,321 kg/ha, respectively.  During the last three years of the study
amounts of solids applied each year were higher than in previous years, which
resulted in concomitantly higher loading rates of sludge-borne metals.  But
overt symptoms of phototoxicity were never observed in corn.

   As can be seen in Table 16, organic-C and total N were significantly
increased in the 0 to 15 cm depth of strip-mine spoil as a result of annual
sludge applications.  But both remained fairly constant during the last three
years of the study, indicating that an equilibrium between annual additions
and decompositional losses of organic matter had been established.  The
equilibrium in spoil was attained at an organic matter content of about 1%
less than in Blount silt loam treated with similar annual applications of
sludge.

                                       95

-------
TABLE 14.  CONCENTRATIONS AND TOTAL ANNUAL AMOUNTS OF SEVERAL CONSTITUENTS
           CONTAINED AT RELATIVELY HIGH CONCENTRATIONS IN SLUDGES APPLIED ON
           PLOTS OF STRIP-MINE SPOIL MATERIAL PLANTED TO CORN.

Year


1978
1979

1978
1979
Solids
%



4.39
4.69
mt/lia
64.3
79.6
local
N



2215
2128
NH4-N



971
942
1' K
Annual means
i i f »

1410 250
1400 211
Na



122
108
Ca



1630
1650
Mg



579
597
Fe



1960
2130
Total amounts applied annually

3378
378fa

1431
1680

2150 381
2480 375

186
192

2480
2930

883
1060

2990
3790

TABLE  15.  CONCENTRATIONS AND TOTAL ANNUAL AMOUNTS OF SEVERAL METALS IN
           SLUDGES APPLIED ON PLOTS OF STRIP-MINED SPOIL MATERIALS PLANTED
           TO CORN.

Year
1978
1979
1978
1979
Mn

21.4
21.7

32.6
38.6
Zn

219
209

334
372
Cd

15.2
14.9
Total
23.2
26.5
Cu Ni Cr
A »» 1 /I
86.8 23.4 170
77.3 21.1 170
amounts applied annually
132 35.7 259
138 37.5 302
Pb

57.2
57.9

87.2
103
                                     96

-------
TABLE 16.  TOTAL N,  ORGANIC-C, pH,  AND TOTAL CONCENTRATIONS OF MACRO-
           CLEMENTS AND MINOR ELEMENTS IN 0 TO 15, 15 TO 30, 30 TO 46, AND
           61 TO 76 CM DEPTHS OF STRIP-MINED SPOIL MATERIAL IN PLOTS LOCATED
           IN FULTON COUNTY, ILLINOIS, a/b/

Par am- Depth
eter cm
Organic-
C 0-15
15-30
30-46
61-76
N 0-15
15-30
30-46
61-76
Year
1978
1979
1973
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979

0

0.33
0.40
0.19
0.20
0.15
0.16
0.15
0.14
0.067
0.058
0.045
0.048
0.036
0.043
0.040
0.038
Sludge
1/4
Max

0.59
0.60
0.18
0.16
0.16
0.16
0.14
0.14
0.084
0.088
0.044
0.045
0.041
0.043
0.042
0.035
application r,ates
1/2
Max

0.94
0.81
0.22
0.21
0.15
0.15
0.16
0.15
0.110
0.105
0.043
0.045
0.043
0.033
0.043
0.040
Max

1.22
1.36
0.29
0.30
0.18
0.21
0.18
0.15
0.144
0.153
0.055
0.055
0.041
0.043
0.041
0.035
LSD

0.33**
0.38**
n. s.
n. s.
n.s.
n.s.
n.s.
n.s.
0.035**
0.026**
n.s.
n. s.
n.s.
n. s.
n.s.
n.s.
                                (continued)

                                     97

-------
TABLE  16.  (concinued)

Param-
eter
P



K



Depth
cm
0-15
15-30
30-46
61-76
0-15
15-30
30-46
61-76
Year
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979

0

0.079
0.087
0.052
0.054
0.035
0.038
0.033
0.045
2.00
1.96
1.85
2.11
1.90
2.22
1.98
2.08
Sludge
1/4
Max

0.088
0.117
0.054
0.059
0.040
0.037
0.041
0.048
2.04
2.05
1.96
2.08
1.98
1.97
2.05
2.12
application rates
1/2
Max
———-—%•
0.142
0.146
0.063
0.071
0.032
0.041
0.033
0.049
1.90
2.00
1.94
2.05
1.95
2.05
2.03
1.84
Max

0.207
0.247
0.068
0.080
0.047
0.039
0.034
0.046
1.97
2.02
1.90
2.06
1.86
1.98
1.90
2.04
LSD

0.059**
0.67**
n. s.
0.011**
n. s.
n.s.
n.s.
n.s.
n. s.
n.s.
n.s.
n. s.
n. s.
n. s.
n. s.
n.s.
                                 (continued)





                                    98

-------
TABLE  16.   (continued)

Sludge application rates
Pa ram- Depth
eter cm Year
Na 0-15 1978
1979
15-30 1978
1979
30-46 1978
1979
61-76 1978
1979
Ca 0-15 1978
1979
15-30 1978
1979
30-46 1978
1979
61-76 1978
1979


0

0.760
0.678
0.659
0.701
0.748
0.732
0.772
0.766
0.902
0.882
0.782
0.834
1.03
0.873
1.31
0.939


1/4
Max

0.626
0.686
0.644
0.645
0.676
0.683
0.724
0.677
0.958
0.901
1.02
1.01
1.01
1.01
1.19
1.08
(continued)
99
1/2
Max
.__ 	 -%-•
0.618
0.700
0.712
0.710
0.765
0.755
0.792
0.657
0.972
0.959
1.00
1.07
0.841
1.09
1.09
0.952


Max

0.715
0.703
0.658
0.774
0.725
0.778
0.825
0.744
1.08
0.946
0.932
0.831
1.00
0.773
1.05
1.09


LSD

n. s.
n.s.
n. s.
n. s.
n. s.
n. s.
n. s.
n.s.
n.s.
n. s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.



-------
TABLE  16.  (continued)

Param-
eter
Mg



Fe



Depth
cm
0-15
15-30
30-46
61-76
0-15
15-30
30-46
61-76
Year
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979

0

0.586
0.777
0.699
0.752
0.829
0.835
0.777
0.836
3.36
3.03
3.24
2.97
3.16
3.75
3.14
3.74
Sludge
1/4
Max

0.731
0.761
0.830
0.808
0.857
0.854
0.965
0.865
3.70
3.50
3.61
3.23
3.63
3.55
3.31
4.18
application rates
1/2
Max
— — — — — %
0.648
0.773
0.759
0.834
0.856
0.918
0.944
0.737
3.55
3.49
3.53
3.07
3.19
3.47
3.17
3.96
Max

0.782
0.758
0.706
0.792
0.887
0.729
0.863
0.781
3.62
3.41
3.21
3.06
3.04
3.23
3.14
3.57
LSD

n. s.
n. s.
n. s.
n.s.
n. s.
0.117*
n.s.
n.s.
n. s.
0.39**
n. s.
n.s.
n.s.
n.s.
n.s.
n. s.
                                  (continued)
                                     100

-------
TABLE  16.   (continued)

Sludge application races
Param-
eter

Mn







Zn







Depth
cm

0-15

15-30

30-46

61-76

0-15

15-30

30-46

61-76

Year

1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
0


686
592
512
510
669
617
622
528
74
82
57
56
60
63
64
61
1/4
Max


736
680
741
631
745
741
824
625
135
150
73
70
65
62
70
70
1/2
Max
/i

638
648
638
600
672
732
634
580
210
208
74
75
66
60
74
62
Max


666
575
569
580
612
572
609
578
286
345
77
95
66
62
69
63
LSD


n. s.
n.s.
n.s.
n. s.
n.s.
n. s.
n.s.
n.s.
69**
80**
?**
26**
n. s.
n.s.
n.s.
n.s.
                                  (continued)




                                     101

-------
TABLE  16.   (continued)

Pa ram- Depth
eter cm Year

Cd 0-15 1978
1979
15-30 1978
1979
30-46 1978
1979
67-76 1978
1979
Cu 0-15 1978
1979
15-30 1978
1979
30-46 1978
1979
61-76 1978
1979



0


1.0
2.0
<0.25
0.41
<0.25
<0.25
<0.25
<0.25
31
30
26
27
29
23
42
23


Sludge
1/4
Max


4.8
6.0
<0.25
0.80
<0.25
0.36
<0.25
<0.25
51
60
33
33
35
20
31
20
(continued)
102
application rates
1/2
Max


7.8
9.7
0.55
1.13
<0.25
0.54
<0.25
0.5^
78
73
41
31
38
23
33
22


Max
/ko— — — ~

15.2
18.3
0.74
2.39
<0.25
0.75
<0.25
0.66
110
118
38
39
50
23
26
24


LSD


5.9**
5.0**
0.35**
1.30*
n.s.
0.36**
n. s.
n.s.
29**
30**
8*
n. s.
n.s.
n. s.
n. s.
n. s.



-------
TABLE 16.   (continued)

Sludge application rates
Param-
eter

Ni







Cr







Depth
cm

0-15

15-30

30-46

61-76

0-15

15-30

30-46

61-76

Year

1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
1978
1979
0

1/4
Max

1/2
Max

Max
/!,-
LSD


39
44
36
42
36
46
33
45
77
72
71
57
73
62
76
56
47
47
38
48
38
40
36
49
126
103
76
59
74
57
76
70
51
53
39
47
36
40
35
45
174
127
83
70
72
62
76
56
57
66
40
61
37
45
34
52
232
162
88
77
77
60
78
61
10*
9**
n.s.
n. s.
n.s.
n.s.
n.s.
n. s.
61**
43**
n.s.
15**
n.s.
n. s.
n.s.
n. s.
                                 (continued)





                                     103

-------
 TABLE  16.  (continued)

Param- Depth
eter cm Year
Pb 0-15 1978
1979
15-30 1978
1979
30-46 1978
1979
61-76 1978
1979

0

14
27
12
12
13
14
9
12
Sludge
1/4
Max

29
33
13
15
12
14
10
13
application rates
1/2
Max
	 	 jno/b
Max

45 68
47 76
15 15
17 32
14 14
14 18
9 9
19 14
LSD

16**
33**
n. s.
n. s.
n. s.
n. s.
n. s.
n.s.

pH 0-15 1978
1979
15-30 1978
1979
30-64 1978
1979
61-76 1978
1979
7.3
7.8
7.2
8.0
7.3
7.9
7.2
7.7
7.5
7.8
7.5
7.9
7.4
7.9
7.4
7.7
7.4 7.2
7.6
7.4
7.8
7.2
7.8
7.3
7.7
7.5
7.4
7.7
7.3
7.7
7.0
7.6
0.2*
0.2**
n.s.
0»2**
n.s.
0.2*
0.3**
n.s.

_§_/  Concentrations are on a dry weight basis.
W  Digested sludge was applied annually on replicated plots at various rates
    and control plots were annually treated with relatively high rates of in-
    organic fertilizer.
*,**  Significantly different at P<0.05 and P<0.01, respectively.
                                     104

-------
   Sludge applications increased Ni and Pb in the 0 to 15 cm,  P,  Cu,  Cr,
and Zn in the 0 to 30 cm,  and Cd in 0 to 46 cm depths of spoil.  Only about
one-third of the total amounts of these elements applied on strip-mine spoil
as constituents of sludge can be accounted for by amounts accumulated in the
76 cm death of spoil.  Amounts of these elements that can be accounted for in
level calcareous spoil are less by about 50% than those accounted for in acid
Blount silt loam.   Concentrations of K, Na, Ca,  Mg, Fe, and Mn in spoil were
not affected by sludge applications.  Maximum sludge applications tended to
decrease the pH of spoil,  although the differences were relatively small and
were not decreased below a pH of 7.

   Grain and stover yields for 1978 are presented in Table 17.  Yields were
not determined in 1979 due to a severe crusting problem and drought con-
ditions that reduced plant populations to less than 9,000 plants/ha.   Al-
though mean yields over all years since the study was initiated shows some
advantage for sludge applications, yields in 1978 were lower on sludge-
amended spoil than on control plots treated with inorganic fertilizer.  In
all years,  yields have been disappointingly low, regardless of treatment.  It
appeared that the main reason for low yields was due to a restricted rooting
depth, resulting in a moisture stress at the critical pollination growth
stage.  Irrigating with liquid sludge only partially alleviated the moisture
stress.

   Data presented in Table 18 shows that concentrations of leaf- Ca during
the last year and leaf- Mg during both years were reduced by sludge applica-
tions.  Concentrations of Mn and Cu were increased in leaves and stover
during both years by sludge applications.  Concentrations of Ni were in-
creased by sludge treatments in grain and stover during both years, but not
in leaf.  Zinc and Cd concentrations were increased in all plant tissues by
sludge applications.  Concentrations of Zn and Cd in corn plant tissues from
calcareous spoil were as high as those found in tissues of the same hybrid
grown on acid Blount.  Furthermore, the accumulations of sludge-borne Cd and
Zn in calcareous spoil by repeated applications of sludge caused increased
levels of these metals in plant tissues, but such accumulations in acid
Blount affected Cd and Zn concentrations in corn to only a minor extent
(Hinesly et al., 1981).


Summary and Conclusion

   Further work is needed to determine 1) why digested sewage sludge failed
to ameliorate the chemical and physical properties of strip-mine spoil that
adversely affects the growth of row crops, 2) why some trace elements ap-
peared to be more available for plant uptake, 3) how chemical elements ap-
plied as constituents of sludges are lost from strip-mined spoil, and 4)
why losses of selected chemical elements from calcareous spoil were higher
than those in acid Blount.
                                      105

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TABLE  17.  PLANT POPULATIONS AND GRAIN AND STOVER YIELDS OF CORN  GROWN ON
            STRIP-MINED SPOIL MATERIAL WITH AND WITHOUT DIGESTED SEWAGE
            SLUDGE IN 1978.


            	Sludge  application rates	
            0         1/4        1/2       Max        LSD
            	Max	Max	

                             Plane Population
            	number/ha	
            47900     54100      46100     47300

                              Grain yields
            	______________rat / ha______	
            4.00      2.82       3.14      3.08       0.83*

                              Stover yields
            •__••_»«—_ __*«•«_>^B^_»B«_««—*• «^a™mt / n__"-~™^~»~-—»•—>•—>^~»^^~*~~~"~™~—'~"^^^~»~"~"
            5.52      5.74       6.66      7.07       n.s.
                                       106

-------
TABLE 18.  CONCENTRATIONS OF SELECTED CHEMICAL ELEMENTS IN CORN PLANT TISSUES
           FROM PLOTS OF STRIP-MINED SPOIL MATERIAL WITH AND WITHOUT SLUDGE.a/

Sludge application rates
Constit-
uent

S




P




Year

1978
1979

1978
1979

1978
1979
1978
1979

1978
1979

1978
1979
0


3.38
2.77

1.66
1.91

1.07
1.66

3130
3030

3200
2770

940
1730
1/4
Max


3.28
2.81

1.58
1.66

1.11
1.20

3140
2330

3280
2440

1070
1480
1/2
Max

%____ 	
Leaf
3.21
2.78
Grain
1.74
1.86
Stover
1.27
1.43
Leaf
3060
2440
Grain
3370
2640
Stover
1150
1450
Max


3.49
3.20

1.90
1.72

1.55
1.66

3020
2900

3570
2450

1590
1600
LSD


n. s.
n.s.

n.s.
n.s.

0.31*
n.s.

n.s.
517**

n.s.
n.s.

n.s.
n.s.
                                 (continued)

                                     107

-------
TABLE 18.  (continued)

Sludge application rate
Constit-
uent

Ca





Mg




Year

1978
1979

1978
1979 .

1978
1979

1978
1979

1978
1979

1978
1979
0


7700
8530

67
61

3300
4940

3370
4090

1300
1300

2440
3340
1/4
Max


6950
7940

70
44

3480
5200

3210
3270

1410
1220

2810
3260
1/2
Max
1 1
Leaf
7770
7240
Grain
65
56
Stover
3430
5140
Leaf
2940
3170
Grain
1390
1320
Stover
2880
3080
Max


8320
5820

81
59

3840
5520

2760
2540

1370
1160

2710
2920
LSD


821*
1950**

n.s.
n.s.

n.s.
n.s.

419**
879*

n. s.
n.s.

n.s.
n.s.
(continued)



108




-------
TABLE 18.  (continued)

Sludge application rates
Constit-
uent Year

Fe 1978
1979

1978
1979

1978
1979

Mn 1978
1979

1978
1979

1978
1979
0


119
204

21
25

375
390

81
77

6
8

57
61
1/4
Max


123
176

19
20

184
406

64
61

6
6

55
73
1/2
Max
/i ff
Leaf
124
204
Grain
21
23
Stover
234
422
Leaf
96
87
Grain
6
7
Stover
62
81
Max


129
157

22
24

158
362

155
160

6
8

90
104
LSD


n. s.
n. s.

n. s.
n.s.

n. s.
n.s.

32**
41**

n.s.
n.s.

21**
23*
                                 (continued)




                                    109

-------
TABLE 18.   (continued)

Sludge application rates
Constit-
uent Year

Zn 1978
1979

1978
1979

1978
1979

Cd 1978
1979

1978
1979

1978
1979
0


48
65

27
20

32
41

1.91
3.66

0.160
0.146

2.14
4.78
1/4
Max


114
100

37
24

97
106

7.20
10.3

0.280
0.293

7.70
14.0
1/2
Max
/i
Leaf
201
197
Grain
44
30
Stover
183
230
Leaf
16.9
20.6
Grain
0.410
0.603
Stover
16.2
31.8
Max


317
346

51
33

345
338

33.7
37.0

0.830
1.12

40.0
68.8
LSD


72**
67**

7**
6**

47**
103**

7.63**
5.75**

0.12**
0.53**

9.54**
16.1**
                                 (continued)
                                    110

-------
TABLE  18.  (continued)

Sludge application rates
Constit-
uent Year

Cu 1973
1979

1978
1979

1978
1979

Ni 1978
1979

1978
1979
1978
1979
0


8
10

1.7
2.3

A
6

<0.6
<0.6

<0.6
<0.6
<0.6
<0.6
1/4
Max


8
9

1.7
2.0

5
7

<0.6
<0.6

0.8
<0.6
<0.6
0.8
1/2
Max
/ljr»
Leaf
10
12
Grain
1.8
2.3
Stover
6
10
Leaf
<0.6
<0.6
Grain
1.0
<0.6
Stover
0.9
1.1
Max


11
18

1.6
2.0

6
13

0.7
0.9

2.0
2.0
1.6
1.9
LSD


1**
5**

n. s.
n. s.

!_**
2**

n. s.
n. s.

0.8**
0.6**
0.5**
0.8*
(continued)


111




-------
TABLE 18.   (continued)

Sludge application rates
Constit-
uent Year 0


Cr 1978 0.8
1979 <0.1

1978 0.3
1979 <0.1

1978 1.1
1979 1.2

Pb 1978 0.8
1979 0.7

1978 <0.6
1979 <0.6

1978 2.1
1979 1.4
1/4
Max


0.3


-------
                                    SECTION 5

                                 SPECIAL STUDIES
CHANGES IN STRIP-MINE SPOIL CHARACTERISTICS AND RESPONSE OF PLANTS  TO HIGH-
RATE SEWAGE SLUDGE APPLICATIONS

Introduction

   Methods of applying sludges on strip-mined lands that are consistent
with crop production and erosion control are limited.   Irrigation of growing
crops is limited to systems that apply sludge below the crop canopy.  Spray
irrigation systems cannot be used because leaf surfaces are coated  with
sludge solids that reduce light absorption and,  thus,  photosynthetic pro-
duction rates.  Where stoniness is a problem, subsurface interjection cannot
be used without incurring costly repairs.  Perhaps the worst system yet de-
vised for applying sludges are those involving the use of disc  plows for in-
corporation.  Disc plows cause subsurface compaction that exacerbates the low
infiltration capacity of the predominantly weathered shale and/or glacial till
material.  This leads to increased runoff of water, with concomitant in-
creases in rates of erosion.

   The main objective in reshaping the surface of strip-mined lands should
be to drain off excess water at a non-erosive rate.  Sludge should  be applied
at rates and by methods that maximize the potential benefits of its organic
matter contents to ameliorate physical properties of spoil that adversely
affect the growth of plants.  The technology needed to do this  is available.
Level-ridge terraces, equipped with surface inlets, can be used to  control
erosion.  Sludge, dewatered to about 70% moisture, can be applied with ordi-
nary farm manure spreaders.

   On agricultural lands, maximum sludge loading rates should be regulated
according to the potential for contaminating subsurface water supplies with
nitrate-nitrogen.  But on strip-mined lands that have subsided  to form a
compact structureless mass, water movement is too slow through  such material
to present a pollution hazard to ground water supplies.  Protection of sur-
face waters is the main concern and can be accomplished by controlling run-
off waters with structures and monitoring quality of water in impoundments
prior to its release.  Maximum loading rates of dewatered sludge on strip-
mined lands should be limited by the tolerance of plants to sludge  con-
stituents and their effects on crop quality.

   To identify plants that would rapidly establish vegetative cover and
cropping systems that minimize erosion losses from sludge-amended spoil


                                      113

-------
and to compare effects of a large single application with an equal amount
of sludge applied in increments,  a one-time,  relatively high,  sludge-loading
rate study was established on strip-mined spoil banks in Fulton County, Illinois.

Methods and Materials

   An experimental site, with good surface drainage, was selected on spoil
banks which had been in place for about 30 years.  The spoil materiil had a
silty clay loam texture, CaCO. equivalent of 3.2%,  and pH value of 7.5.
Replicated (three) plots, having the dimensions of 21 x 18 m,  were treated
with 0 (control), 224, 448, and 896 mt/ha (dry weight equivalent) of digested
sewage sludge that had an average moisture content of 45%.  Control plots
received 123 kg/ha of N, P.O., and K?0 each year prior to seeding wheat and
rye.  Following the application of sludge and its incorporation with a rotary
plow, each main plot was subdivided into nine plots of 3 x 6 m and two ad-
ditional plots of 6 x 18.2 m.  Each of the smaller subplots were seeded with
one of the following grasses:  big bluestem (Andropogon gerandi). orchard
grass (Dactylis glomerata), perennial ryegrass (Lolium perenne), redtop
(Agrostis alba), reed canarygrass (Phalaris arundinacea), smooth brome
(Bromus inermis), tall fescue (Festuca elatior).  timothy (Phleum pratense),
and western wheatgrass (Agropyron smithii).  Rye (Secal cereale) or wheat
(Triticum vulgare) was seeded on the two larger subplots.  During the first
week of May, 3 m wide strips of rye and wheat were killed with paraquat and
corn (Zea mays) was planted in the dead residues with a no-till planter.

   From strips (3 x 18.2 m) of wheat and rye that were not sprayed with
paraquat the top four leaves from 150 randomly selected plants were collected
just before head emergence.  Grain and straw samples were collected at the
time of harvest.  The leaf adjacent to the primary ear shoots  was collected
from ten corn plants in each of the dead wheat and rye plots when about 10%
of the plants had tasseled.  The leaves were washed with distilled water,
dried at 60 C and ground in a Wiley mill.  Corn grain and stalk samples were
collected at the time of harvest.

   Samples of spoil were collected from all main plots before sludge ap-
plications were made and each spring from subplots after sludge was applied.
Six 2.5 cm diameter samples were collected from each subplot with stainless
steel tubes to a depth of 91 cm and composited by 15 cm depth  increments,
except the lower increment was 30 cm.  Additional samples of the surface
15 cm depth were collected periodically for determination of organic carbon,
nitrogen, conductivity of saturated extracts, and changes in physical properties.

   Sludge and sludge-amended spoil samples were analyzed for C, N,  P, K, Mg,
Ca, Na, Fe, Mn, Zn, Cu, Cr, Pb, Ni, and Cd concentrations.  Plant tissues
were analyzed for contents of the same elements except C, K, Na.  Chemical
analyses were the same as those described for studies discussed previously
in this report and the procedures for measuring physical properties were
those described in Agronomy Monograph No. 9 (Black et al., 1965).

Results and Discussion

Chemical and Physical Properties of Spoil—

                                      114

-------
   Concentrations of the several chemical elements of interest in sludge
are shown in Table 19,  along with concentrations in the 0-15 cm depth of
sludge-amended spoil where various amounts of sludge were incorporated.
Except for Mn, concentrations of all chemical elements were significantly
affected by sludge applications.  Potassium and Na concentrations in spoil
were decreased by sludge applications while concentrations of all other
selected elements were increased.  Although the highest sludge application
markedly increased organic-C and N concentrations in spoil, C to N ratios
were changed only slightly from 10.9 to 10.3.  The C to N ratio of sludge
itself is intermediate between control plots and maximum-sludge-amended
spoil.  Two years after sludge was applied no significant changes in organic-
C and total N concentrations in spoil were observed, indicating that the
sludge organic matter was highly stabilized against further degradation,
even before it was applied.  Phosphorus and Fe contents were somewhat higher
in the sludge used in this study, which was dredged from a storage reservoir,
as compared to sludge drawn directly from anaerobic digesters at the same
Chicago wastewater treatment plant.  However, concentrations in sludge samples
were not high enough to account for the concentrations of these two elements
found in spoil amended with 896 mt/ha of sludge.  During storage P and Fe
may have been concentrated by precipitation and sedimentation processes,
but samples of dewatered sludge collected from manure spreaders evidently
did not contain concentrations as high as those actually applied.

   Concentrations of selected chemical elements, present in strip-mine
spoil to a depth of 90 cm, before sludge was applied are shown in Table 20.
In comparison to these data, it can be seen in Table 21 that concentrations
of C, Cr, Ni, and Zn were increased in the 15 to 30 cm depth after sludge
was applied.  Two years after sludge was applied, Cu concentrations in 15
to 30 cm and 30 to 45 cm depths had been increased by sludge application.
Concentrations of Cd and Pb were increased at all depths above 60 cm with
increasing sludge loading rates.  Concentrations of Mg were increased at all
sampling depths above 90 cm.  At a depth of 15 to 30 cm, concentrations of
chemical elements reflect those added as constituents of sludge which was
incorporated in the upper portion of this zone, as a result of mixing with
the rotary plow. . But, apparently Cd, Cu, and Pb had migrated to deeper
zones by some process.  Migration may have been by leaching or translocation
by invertebrates and/or plant roots.

   After sludge was incorporated with spoil, the surface pH (0 to 18 cm
depth) was reduced from 7.5 to 7.0, 6.3, and 6.0 by 224, 448, and 896 mt/ha
of sludge solids, respectively.  These pH values remained fairly constant
after incorporation.  The pH of water saturated extracts, from spoil with
and without sludge, are shown in Figure 1.  These extract were obtained to
determine differences in electrical conductivities associated with various
sludge loading rates and these results are shown in Figure 2.  Electrical
conductivities ranged from 2.2 mmho/cm in spoil samples from control plots
to 6.6 mmho/cm in spoil amended with 896 mt/ha of sludge.

   The results obtained by standard methods of determining aggregate sta-
bilities by wet sieving with the Yoder apparatus for 10 minutes are shown in
Figure 3.  Water stable aggregates greater than 0.25 mm increased from 12.2%
in samples from control plots as compared to 42.1% in maximum sludge-amended
spoil.
                                      115

-------
Table 19 .  Concentrations of selected  chemical elements in  sludge  and  the
           0-15 cm depth of sludge-amended  spoil  bank material.  Beginning
           in October of 1979,  separate  spoil samples were  taken from  plots
           planted to grasses  (Gr) and corn (Co).
Spoil (0-15 cm)
Sludge Application
Element

Org.-C








N








P








Sampling
Sludge date

12.20 7-78
10-78
4-79
10-79 (Gr)
10-79 (Co)
3-80 (Gr)
3-80 (Co)
7-80 (Gr)
7-80 (Co)
1.17 7-78
10-78
4-79
10-79 (Gr)
10-79 (Co)
3-80 (Gr)
3-80 (Co)
7-80 (Gr)
7-80 (Co)
3.60 7-78
10-78
4-79
10-79 (Gr)
10-79 (Co)
3-80 (Gr)
3-80 (Co)
7-80 (Gr)
7-80 (Co)

0


1.38
1.40
1.11
1.50
1.49
1.16
1.12
1.59
1.58
0.127
0.125
' 0.102
0.125
0.131
0.103
0.093
0.120
0.094
0.044
0.083
0.080
0.080
0.079
0.075
0.067
0.087
0.072

224
	 
-------
Table 19.   Concentrations of selected chemical elements in sludge and the
continued  0-15 cm depth of sludge-amended spoil bank material.  Beginning
           in October of 1979, separate spoil samples were taken from plots
           planted to grasses (Gr) and corn (Co).
                                               Spoil (0-15 cm)
                                           Sludge Application Rates
                                         	mt/ha-
                 Sampling
Element  Sludge    date
224
448
896
n.s. = not significant.
* = significant at 0.05
-------
 Table 19 .   Concentrations of selected chemical elements in sludge and the
continued   0-15 cm depth of sludge-amended spoil bank material.  Beginning
            in October of 1979,  separate spoil samples were taken from plots
            planted to grasses (Gr) and corn (Co).
Spoil (0-15 cm)
Sludge Application Rates
Sampling
Element Sludge date

0

_•• /u.
224 448

896

LSD
v t A -i v»*- ^

Mg 1.70 7-78
10-78
4-79
10-79 (Gr)
10-79 (Co)
3-80 (Gr)
3-80 (Co)
7-80 (Gr)
7-80 (Co)
Fe 6.85 7-78
10-78
4-79
10-79 (Gr)
10-79 (Co)
3-80 (Gr)
3-80 (Co)
7-80 (Gr)
7-80 (Co)

Mn 817 7-78
10-78
4-79
10-79 (Gr)
10-79 (Co)
3-80 (Gr)
3-80 (Co)
7-80 (Gr)
7-80 (Co)
0.795
0.802
0.980
0.919
0.954
0.995
1.04
0.949
0.991
4.48
3.76
3.76
3.76
3.93
3.61
3.75
4.07
3.87


630
629
548
598
581
403
435
556
539
1.02
0.809
1.02
1.02
1.02
1.02
0.997
0.537
0.862
4.94
4.68
4.48
4.94
4.94
4.46
4.36
4.57
4.49
.

636
600
654
602
585
301
398
565
615
1.05
1.10
1.13
1.03
1.14
1.01
1.07
1.00
0.926
6.00
6.44
6.32
6.17
5.82
5.66
5.30
5.48
5.12
(dry weight)
621
662
652
619
615
353
445
592
598
1.40
1.24
1.29
1.30
1.28
1.28
1.26
1.08
0.988
8.08
8.37
8.26
9.10
8.48
7.84
7.81
7.03
6.56


722
677
625
648
618
356
366
639
601
0.371**
0.249*
0.168**
0.151**
0.233**
0.201*
0.169**
0.166**
n. s.
2.22**
0.84**
1.75**
1 . 50**
2.07**
2 . 09**
1.18**
1.16**
0.74**


n.s.
n.s.
n.s.
n.s.
n.s.
17. 8**
n.s.
n.s.
n.s.
 n.s.  • not significant.
 * = significant at 0.05
-------
 Table 19 .   Concentrations of selected chemical elements in sludge and the
continued   0-15 cm depth of sludge-amended spoil bank material.  Beginning
            in October of 1979,  separate spoil samples were taken from plots
            planted to grasses (Gr) and corn (Co).
Sooil (0-15 cm)


Sampling
Element Sludge date
Zn 4230 7-78
10-78
4-79
10-79
10-79
3-80
3-80
7-80
7-80
Cd 276 7-78
10-78
4-79
10-79
10-79
3-80
3-80
7-80
7-80
Cr 2760 7-78
10-78
4-79
10-79
10-79
3-80
3-80
7-80
7-80
•

(Gr)
(Co)
(Gr)
(Co)
(Gr)
(Co)



(Gr)
(Co)
(Gr)
(Co)
(Gr)
(Co)



(Gr)
(Co)
(Gr)
(Co)
(Gr)
(Co)
Sludge ADplication Rates

0

99.
87.
84.
94.
90.
94.
94.
133
91.
0.
0.
0.
0.
0.
0.
0.
2.
0.
41.
47.
43.
63.
50.
53.
52.
54.
47.



3
6
8
9
2
1
2

9
545
307
715
793
469
943
484
29
435
9
6
6
6
0
8
4
5
9

224

rag/
503
627
729
991
750
611
475
392
440
28.
36.
42.
48.
45.
31.
22.
23.
22.
206
215
390
396
385
310
235
168
177


kg







6
4
5
4
9
2
2
6
5









_mf /Vi-*_
448


(dry weigh
884
1470
1530
1460
1350
1380
993
1040
905
59.
98.
95.
96.
84.
70.
56.
59.
56.
430
566
847
848
713
625
529
449
360







3
5
4
5
8
4
4
1
0










896

O ______
1660
2550
2500
2770
2510
2270
2300
1690
1570
140
157
161
171
163
136
142
110
99.0
1130
994
1420
1550
1240
1230
1280
785
720

LSD

603**
321**
750**
595**
594**
954**
429**
431**
382**
50.3**
19.4**
36.5**
39.1**
34.7**
63.5**
25.0**
34 . 0**
25.3**
408**
152**
356**
487**
363**
551**
261**
259**
204**
n.s. = not significant.
* = significant at 0.05
-------
 Table 12   Concentrations of selected chemical elements in sludge and the
 continued   0-15 cm depth of sludge-amended spoil bank material.   Beginning
            in October of 1979,  separate spoil samples were taken from plots
            planted to grasses (Gr)  and corn (Co).
Spoil (0-15 cm)
Sludge Application Rates
Sampling
Element Sludge date
-
Cu 1380 7-78
10-78
4-79
10-79 (Gr)
10-79 (Co)
3-80 (Gr)
3-80 (Co)
7-80 (Gr)
7-80 (Co)
Ni 284 7-78
10-78
4-79
10-79 (Gr)
10-79 (Co)
3-80 (Gr)
3-80 (Co)
7-80 (Gr)
7-80 (Co)
Pb 1090 7-78
10-78
4-79
10-79 (Gr)
10-79 (Co)
3-80 (Gr)
3-80 (Co)
7-80 (Gr)
7-80 (Co)

0


36.6
29.9
26.5
32.9
34.1
32.6
32.4
35.0
30.1
35.8
39.9
44.4
30.4
34.5
40.4
38.3
45.7
43.8
13.0
13.5
15.0
18.1
18.8
18.7
16.0
23.5
12.5

224
.,

174
192
213
244
250
190
144
144
139
62.6
79.4
88.8
64.6
72.0
70.7
69.4
61.9
65.7
122
145
134
172
178
140
100
105
103
/v
448
(dry weigl
345
463
431
478
424
348
299
333
290
99.5
129
138
114
100
112
105
103
93.2
247
357
284
374
318
269
226
245
212

896
,r\

684
746
593
828
766
661
680
544
521
176
201
203
172
173
194
196
153
141
548
676
476
641
604
540
538
389
389

LSD


158**
101**
158**
174**
152**
273**
109**
135**
115**
54.1**
22.8**
37.6**
31.3**
30.3**
64.6**
24.8**
36.4**
19.7**
178**
232**
145**
127**
111**
251**
104**
121**
111**
n.s. - not significant.
* = significant at 0.05
-------
Table 20.  Concentrations of selected chemical elements and pit of strip-mine spoil materials in
           0- to 15-, 15- to 30-, 30- to 46-, 46- to 61-, and 61- to 91-cm depths calculated
           prior to sludge application.  These data are means of composite samples taken from
           three replicated blocks of four experimental plots each.
Strip-Mine Spoil Depths (cm)
Analyte
org.-C
N
K
Na
Ca
Mg
Fe
Mn
Cd
Cr
Cu
Ni
Pb
Zn
P
pll
0-15

0.95
0.089
2.23
1.04
0.790
0.980
3.67
589
0.31
44.0
29.6
32.7
9.50
98.5
583
7.4
15-30

0.51
0.073
2.23
0.823
0.692
0.780
3.65
551
0.521
46.0
28.4
33.9
8.21
96.8
488
7.4
30-46

%____—.
0.51
0.059
2.27
1.04
0.739
0.932
3.86
602
<0.25
44.0
36.7
32.7
10.2
99.5
494
7.6
46-61

0.62
O.Q60
2.29
1.12
0.619
0.993
3.64
664
<0.25
40.2
43.2
27.6
11.2
98.5
498
7.5
61-91

0.82
0.071
2.25
1.07
0.874
0.966
3.93
614
<0.25
46.3
62.8
35.1
9.70
104
572
7.6

-------
Table 21.  Concentrations of selected chemical elements  in  sludge-amended
          spoil bank materials in 0- to 15-r, 15-  to  30-, 30-  to  46-,  46- to
          61-, and 61- to 91 cm depths.  -  ^'

Analvte
Org.-C




N




P




Depths
cm
0-15
15-30
30-46
46-61
61-91
0-15
15-30
30-46
46-61
61-91
0-15
15-30
30-46
46-61
61-91
Plot
Type
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass


0

1.58
1.59
0.685
0.436
1.03
0.456
0.568
0.482
0.468
0.558
0.094
0.120
0.063
0.066
0.058
0.068
0.065
0.063
0.061
0.062
0.072
0.087
0.029
0.080
0.029
0.029
0.030
0.029
0.030
0.033
Sludge

':24
q
2.32
2.41
0.839
0.484
1.11
0.393
0.524
0.462
0.446
0.624
0.172
0.175
0.078
0.084
0.063
0.065
0.064
0.042
0.058
0.063
0.415
0.501
0.052
0.136
0.058
0.048
0.061
0.053
0.046
0.045
Application Rates

448
I (dry weigl
2.83
3.26
0.784
0.664
1.22
0.420
0.636
0.534
0.566
0.432
0.258
0.305
0.085
0.093
0.085
0.047
0.069
0.050
0.063
0.058
0.996
1.24
0.070
0.211
0.066
0.053
0.049
0.035
0.039
0.037

896
Tt-1 _______
4.81
4.81
1.02
1.28
0.936
0.568
0.559
0.634
0.582
0.591
0.427
0.482
0.108
0.113
0.074
0.063
0.068
0.067
0.069
0.069
2.07
2.19
0.072
0.454
0.063
0.057
0.043
0.048
0.048
0.045

LSD

1.48**
1.47**
n.s.
0.582**
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
0.108**
0.118**
0.023**
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
0.438**
0.523**
n.s.
0.170**
0.027*
n.s.
0.019*
0.014*
n.s.
n.s.
                                 (continued)
                                     122

-------
Table 21.  (continued)

Analyte
K




Na




Ca




Depths
cm
0-15
15-30
30-46
46-61
61-91
0-15
15-30
30-46
46-61
61-91
0-15
15-30
30-46
46-61
61-91
Plot
Type
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass


0

2.31
2.20
2.30
2.08
2.28
2.25
2.19
2.26
2.34
2.20
0.898
0.900
0.932
0.759
0.906
0.891
0.883
0.915
0.901
0.820
0.692
0.600
0.660
0.684
0.833
0.737
0.790
0.727
0.862
0.689
Sludge

224
	 	 	 	 __2
2.01
1.91
1.84
1.85
1.90
1.88
1.81
1.71
1.92
1.82
0.754
0.563
0.577
0.542
0.620
0.645
0.531
0.547
0.575
0.597
1.08
0.549
0.666
0.502
0.652
0.505
0.452
0.534
0.502
0.576
Application
	 mt/ha 	
- - 448
(dry weight)-
1.96
1.93
2.14
2.12
2.21
2.25
2.18
2.22
2.22
2.14
0.705
0.750
0.747
0.744
0.823
0.822
0.727
0.749
0.757
0.793
1.20
1.16
0.699
0.721
0.756
0.724
0.692
0.775
0.757
0.685
Races

896

1.60
1.64
2.15
2.09
2.16
2.22
2.13
2.19
2.250
2.150
0.612
0.629
0.857
0.778
0.807
0.842
0.777
0.813
0.748
0.729
1.64
1.60
0.674
0.770
0.656
0.660
0.673
0.687
0.781
0.620


LSD

0.336*
0.254**
0.239**
0.173*
0.202**
n.s.
0.105**
0.192**
0.223*
n.s.
0.189*
0.185**
0.109**
n.s.
0.160**
n.s.
0.109**
0.129**
0.197**
0.144*
0.439*
0.350**
n.s.
n.s.
n.s.
0.122*
n.s.
n.s.
n.s.
n.s.
                                 (continued)
                                     123

-------
Table  21. (continued)

Analvte
Mg




Fe



Mn




Depths
cm
0-15
15-30
30-46
46-61
61-91
0-15
15-30
30-46
46-61
61-91
0-15
15-30
30-46
46-61
61-91
Plot
Type
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass


0

0.991
0.949
0.990
0.982
1.030
0.981
0.994
0.998
1.02
0.901
3.870
4.070
3.990
4.050
3.720
3.810
3.880
3.850
4.330
3.810
539
556
544
559
544
579
561
567
608
570
Sludge

224

0.862
0.537
0.520
0.486
0.560
0.583
0.468
0.423
0.523
0.530
4.49
4.57
3.82
3.97
3.87
3.44
3.97
3.66
4.01
3.63
615
565
542
489
529
500
499
582
556
518
Application Rates

-- 448
(dry weigh!
0.926
1.00
0.840
0.833
0.891
0.938
0.831
0.892
0.796
0.906
5.12
5.48
4.05
4.34
3.96
4.24
4.37
4.08
4.28
3.85
/kg (dry we]
598
592
539
668
568
594
589
605
593
531

896
. \ 	
- ) ————————
0.988
1.080
0.936
0.905
0.844
0.883
0.850
0.892
0.857
0.792
6.56
7.03
4.01
4.31
3.98
3.79
3.66
4.00
4.26
3.79
i rrh r 1 - 	 _—
601
639
589
571
571
544
581
632
676
616

LSD

n. s.
0.166**
0.218**
0.129**
0.061**
0.271**
0.110**
0.235**
0.175**
0.235**
0.739**
1.160**
n.s.
n.s.
n.s.
n.s.
n.s.
n. s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
46*
n.s.
n.s.
n.s.
n.s.
                                  (continued)
                                      124

-------
Table 21.  (continued)

Sludge Application Rates
Depths
Analvte cm
Zn 0-15
15-30
30-46
46-61
61-91
Cd 0-15
15-30
30-46
46-61
61-91
Cr 0-15
15-30
30-46
46-61
61-91
Plot
Type
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass

0

91.9
133.0
88.5
96.0
90.5
92.5
92.7
89.2
103.0
85.8
0.435
2.29
<0.25
0.545
<0.25
0.273
<0.25
0.274
0.371
0.294
47.9
54.5
38.0
41.9
49.1
49.2
45.3
44.5
49.3
46.7

224

440
392
135
195
102
141
100
86.5
85.9
83.2
22.5
23.6
2.98
4.38
1.15
0.986
0.741
0.543
<0.25
<0.25
177
168
55.0
55.0
52.9
50.2
50.8
45.8
44.6
45.0
mt/ha 	
- 448
(dry we
905
1040
162
210
180
118
109
93.0
97.9
81.2
56.0
59.1
4.67
6.81
5.96
1.64
1.63
0.670
0.650
<0.25
360
449
75.9
90.4
87.2
57.1
58.4
56.6
56.3
54.2

896
J rvVif ^ ______
1570
1690
219
410
127
128
78.6
87.8
77.3
78.6
99.0
110.0
11.7
21.5
5.08
4.16
1.51
1.94
1.48
1.30
720
785
95.5
153
69.6
69.4
53.3
54.5
54.8
51.0

LSD

382**
431**
77.2*
148**
n.s.
n.s.
n. s.
n.s.
n.s.
n.s.
25.3**
34.0**
4.44**
8.45**
n.s.
1.98**
n.s.
0.924*
0.921*
n.s.
204**
259**
37.8**
40.9**
n.s.
12.6*
n.s.
n.s.
7.6*
n.s.
                                  (continued)
                                      125

-------
Table  21. (continued)

Sludge Application Rate
Depths
Analyte cm
Plot
Tvpe

0

224
	 rag/kg
Cu 0-15
15-30
30-46
46-61
61-91
Ni 0-15
15-30
30-46
46-61
61-91
Pb 0-15
15-30
30-46
46-61
61-91
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
30.1
35.0
27.4
27.1
28.7
25.8
25.8
25.2
27.9
27.8
43.8
45.7
43.2
44.9
43.1
42.7
43.2
45.1
45.3
41.3
12.5
23.5
12.3
14.4
11.0
11.7
12.6
11.8
12.1
7.86
139
144
36.6
42.4
32.8
30.1
32.0
27.7
26.5
26.7
65.7
61.9
40.9
41.5
38.9
38.7
39.5
37.0
38.2
40.4
103
105
20.4
26.8
13.6
12.1
11.6
11.2
12.0
13.3
nt/ha-
. 448
(dry
290
333
50.1
63.9
59.3
34.9
36.8
30.4
38.9
31.2
93.2
103
44.2
46.8
46.7
43.7
41.3
41.2
40.2
37.6
212
245
27.7
42.5
36.8
20.6
18.4
12.9
13.0
10.7

896
weight) 	
521
544
68.6
144.
43.7
42.0
35.1
32.9
33.0
31.4
141
153
47.4
60.3
41.4
43.9
39.1
38.9
41.1
38.7
389
389
49.4
93.2
25.0
24.5
17.0
18.1
14.3
12.0

LSD
	
115**
135**
19.7**
25.1**
n.s.
8.2**
n.s.
n.s.
n.s.
n.s.
19.7**
36.4**
3.6*
12.8**
n.s.
n.s.
n.s.
3.9**
4.5*
n.s.
Ill**
121**
21.7**
27.6**
n.s.
9.7*
n.s.
4.7*
n.s.
n.s.
                                  (continued)
                                     126

-------
Table 21.  (continued)


J* «1
Analvte cm

pH 0-15
15-30
30-46
46-61
61-91



PI /tf
Type

Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass
Corn
Grass




0

7.57
7.55
7.62
7.40
7.24
7.68
7.62
7.35
7.35
7.60


Sludge

224

7.43
7.41
7.69
7.66
7.68
7.59
7.60
7.53
7.71
7.61


Aoolication

- - 448

— pn units ™
6.96
6.95
7.44
7.32
7.23
7.28
7.35
7.48
7.41
7.40


Rate

896

6.44
6.42
7.16
7.18
7.31
7.39
7.44
7.37
7.42
7.25




LSD

0.74**
0.62**
0.30**
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.

—  Sludge was applied to plots in summer of 1978.

—  Samples were taken from corn and grass plots  in 1980.

*    = Significant at P£0.05.
**   = Significant at P<0.01.
n.s. = Not significantly different.
                                     127

-------
      8.0
 u
 O
 W
U
LU
•o
"5
 3
B
      7.0
      6.0
                    224
                                448
672
896
                     Sludge Applied  (mt/ha)
Figure 1.   pll of Saturated  Extracts from Strip-Mined Spoil Without and
           With Various  Amounts of Incorporated Sewage Sludge.
                             128

-------
 O
 _=
 O
 VJ
                     224
448
672
896
                        Sludge Applied  (mt/ha)
Figure 2.   Electrical Conductivity of Saturated Extracts fron Strip-

           Mined  Spoil Without and With Various Amounts of Incorporated
           Sewage Sludge.


                             129

-------
   Figure 4 shows chat the amounts of water retained at  saturation and
1/3- and 15-bar matric tension increased in proportion to amounts of sludge
applied.  The rate of increase in moisture content at 1/3 bar was higher
than that at 15 bar, thus, there was a small but significant increase in avail-
able moisture holding capacity with the two highest sludge application rates.

Crop Response--
   Corn, wheat, and rye stover and grain yields are exhibited in Table 22.
Corn grain yields on plots treated with low and intermediate sludge loading
rates were significantly higher than those on fertilized control plots and
plots treated with the highest rates of sludge, in the first year.  Corn
stover yields followed a similar pattern.  In the second year, corn grain
yields did not differ by treatment and were considerably less than those
produced during the first year.  But stover yields were  higher in the second
year.  During the second year a severe moisture stress and unusual hot
weather during the pollination period evidently reduced  fertilization to such
an extent that most ears were barren or partially barren of grain.  Differ-
ences in corn yields between wheat and rye mulch were not significant, al-
though the latter provided considerably better coverage  of the spoil sur-
face.  Wheat was preferentially grazed by wild geese for such an extended
period into the spring that grain development was severely reduced.  Rye
was not grazed as intensively as wheat, but yields may have been affected
since there was no significant differences due to treatments.  During the
last year, rye produced significantly higher amounts of  straw on plots treated
with 448 rat/ha of sludge than on control and lesser sludge-treated plots.

   Six weeks after the initial fall seeding of the nine  species of grass,
at a rate of 25 kg/ha, tall fescue and perennial ryegrass were the only ones
present at acceptable stands on most plots.  After seeding again in the
spring, these two were followed by fairly good stands of western wheatgrass.
All grasses were established except big bluestem following the second fall
seeding.  From the standpoint of rapid establishment and vigorous growth on
all sludge-amended plots, tall fescue, perennial ryegrass and western wheat
grass were the best by order of listing.

Inorganic Composition of Plant Tissues—

   Regardless of whether corn was grown in rye or wheat  mulch, higher sludge
loading rates increased concentrations of P, Mn, Cd, and Zn in corn leaves
during both years (Table 23).  Corn leaf-Ni concentrations were increased
by the highest sludge loading rate only during the first year.  Lead concen-
trations in leaves of plants grown in dead wheat were significantly higher
with higher sludge treatments during the first year, but not in the second
year.  It is noteworthy that leaf-Cd concentrations were only about 50%
of those observed during the first year even though the  same hybrid was
planted.  Also, in both years leaf-Cd concentrations were not increased by
doubling the 448 mt/ha sludge application even though Cd concentrations in
spoil were increased from 56 to 142 mg/kg (Table 19).  Zinc contents of leaves
also decreased during the second year, but, unlike Cd, concentrations of Zn
in leaves were directly proportional to amounts of the metal supplied as a
constituent of sludge.
                                      130

-------
 10
 CN
 o
 A
  O)
  
       50
       40
       30
       20
       10
                    224
448
672
896
                     Sludge Applied (mt/ha)
Figure 3.   Percent Water Stable Aggregates in Strip-Mined Spoil
           Without and With Various Amounts of Incorporated Sewaee
           Sludge.
                             131

-------
       J

       X
      •a
       o
      2
      "5
           40 -
           30 -
           20 -
           10 -
                      224
448
672
896
                         Sludge Applied (mt/ha)
Figure 4.  Percent Moisture  Retention  by Strip-Mined Spoil 'Jithout

           and With Various  Anouncs  of Incorporated Sewage Sludge.
                               132

-------
Table 22.  Grain and stover yields for corn, wheat, and rye and plant popula-
          tions for corn on spoil banks with and without sludge.  Corn was
          planted in dead wheat (W) and rye (R) mulch with a no-till planter.
Corn
Year

1979




1980





1979




1980




Sludge Treatment

0
224
448
896
LSD
0
224
448
896
LSD

0
224
448
896
LSD
0
224
448
896
LSD
(W)

2.68
6.77
5.64
3.26
2.70**
1.78
1.59
1.12
0.955
n.s.
	
2.42
6.60
7.20
3.67
3.32**
4.13
7.30
8.15
7.52
n.s.
(R)
——r-r-a-in Yi *»1 Ha
3.29
6.76
6.81
4.20
2.15*
1.91
1.60
1.82
1.41
n.s.
— Stover Yields
2.70
5.75
7.02
3.82
2.13**
3.84
7.42
9.57
7.41
n.s.
Wheat
Cmt/hal 	
\IU L. / iiA/
n.d.
0.72*
1.93b
1.48C
-
n.d.
n.d.
n.d.
n.d.
—
Rye

2.61
2.36
2.41
2.33
n.s.
0.304
0.496
0.918
1.04
n.s.
(mt /ha ) 	
n.d.
n.d.
n.d.
n.d.
—
n.d.
n.d.
n.d.
n.d.
-
n.d.
n.d.
n.d.
n.d.
—
1.59
2.26
4.26
3.96
1.74*
                                    	Plant  Populations  (1000/ha)	
1979



1980



n.d.
a
b
c
n.s.
*
**
0
224
448
896
0
224
448
896
= no data due to bird
= only one plot.
= only two plots .
= three plots.
= not significant.
= significant at P £0
* significant at P £0
31.3
58.2
58.2
36.3
28.9
38.3
43.2
41.0
damage.




.05.
.01.
36.7
50.0
60.2
42.4
28.1
38.3
47.8
41.6







                                      133

-------
        Table 23,   Concentrations of  selected elements in corn leaf from plots with and without sludge.   Corn
                   was planted with a no-till planter In dead wheat and rye.
u>
Planted in Dead Rye
Planted in Dead Wheat
Sludge Application Rates
Element
Mg
Ca
P
N
Zn
Fe
Ma
Year
1979
1980
1979
1980
1979
1980
1979
1980
1979
1980
1979
1980
1979
1980

0

0.48
0.47
0.56
0.68
0.24
0.24
2.81
2.72
51.7
48.5
133
148
25.4
44.0

22^

0.44
0.43
0.66
0.70
0.28
0.29
3.12
2.54
105
67.2
118
120
43.4
37.5

44 8

0.46
0.51
0.71
0.66
0.32
0.29
3.20
3.05
201
106
149
126
72.0
45.8

896

0.58
0.72
0.61
0.72
0.35
0.30
3.25
2.70
276
159
128
136
178
97.2
nil-
/!*«
LSD 0
	 V fHru
0.07*
0.15*
0.07*
n. s.
0.07**
0.03*
0.30*
n.s.
-mg/kg (dry
108**
66*
n.s.
17**
45.4**
34.3**

0.46
0.48
0.61
0.60
0.23
0.25
2.85
2.74
weight)-
50.9
48.4
148
145
25.0
42.4

224

0.47
0.45
0.64
0.70
0.28
0.31
2.96
2.59
92.1
57.5
131
125
42.8
40.0

448

0.48
0.56
0.74
0.78
0.31
0.30
3.13
2.97
184
123
144
126
74.9
53.6

896

0.54
0.72
0.71
0.70
0.36
0.31
3.21
2.79
31'l
155
138
132
179
80.1

LSD

n.s.
0.17*
n.s.
n.s.
0.07**
0.04*
n.s.
n.s.
127**
32**
n.s.
n.s.
44.3**
2.r>.4**
        *,  **  =  significantly  different  at P<0.05  and P_<0.01,  respectively.
        n.s. = no  significant  differences.

-------
Table 23.   Concentrations of selected elements in corn leaf from plots with and without sludge.   Corn
continued  was planted with a no-till planter in dead wheat and rye.
Planted in Dead Rye
Planted in Dead Wheat
Sludge Application Rates
. •« §• y i.«
Element
Ni
Cr
Pb
Cd
Cu
Year
1979
1980
1979
1980
1979
1980
1979
1980
1979
1980
0

<0.62
<0.62
<0.125
0.791
0.70
<0.62
0.334
0.075
12.0
10.8
224

<0.62
<0.62
<0.125
0.150
0.87
<0.62
3.10
3.57
11.8
8.62
448

<0.62
<0.62
<0.125
<0.125
1.30
<0.62
11.0
7.23
12.0
8.91
896

1.80
<0.62
<0.125
0.149
0.92
<0.62
14.5
7.09
13.0
9.18
LSD 0
/kg (dry
0.54**
n. s.
n. s.
n. s.
n. s.
n.s.
6.76**
4.70**
n.s.
n.s.

<0.62
<0.62
<0.125
<0.125
0.74
<0.62
<0.062
<0.062
11. 1
11.2
224

<0.62
<0.62
0.185
0.209
0.92
<0.62
2.29
2.51
11.4
9.11
448

<0.62
<0.62
0.246
0.125
0.94
<0.62
12.2
7.23
11.9
8.75
896

1.58
0.99
0.285
<0.125
1.09
<0.62
16.4
7.05
13.9
9.15
LSD

0.77**
n.s.
n.s.
n.s.
0.18*
n.s.
8.59**'
2.60** -
n.s.
n.s.
*, ** = significantly different at P<0.05 and P^O.Ol,  respectively.
n.s.  = no significant differences.

-------
   In Table 24 it may be seen chat concentrations of Cd,  Ni,  and Zn in
grain were increased by sludge applications each year on  all  plots.  Con-
centration of Mg and P were increased in the first but not the second year.
Grain-Mn concentrations were increased by the highest sludge  loading rate
during both years on plots with rye and the first year with wheat residue
mulches.  Like leaf concentrations, grain-Cd was not increased by doubling
the 448 mt/ha sludge application.  Unlike leaf-Zn, concentrations of Zn in
grain were similar to those of Cd in that the maxinum sludge  loading rate
did not increase concentrations over those produced by the intermediate load-
ing rate.  This may have been due Co the marked reduction in  grain yields
during the second year.

   During both years, sludge applications increased Mg, P, Ni, Cd, Cu and
Zn concentrations in rye grain (Table 25).  Concentration of  N and Fe in rve
grain were increased the first year after sludge was applied.  Although sludge
applications increased Cd concentration in rye grain as compared to those in
grain from control plots, during the second year Cd concentrations were un-
affected by sludge loading rate.  Rye stover had higher concentrations of
Ca, Mg, N, P,  Cd, Ni, and Zn during both years as a result of sludge appli-
cations.  Concentrations of Cr, Fe, and Pb in rye stover  were unaffected by
sludge treatments and Mn concentrations were increased only during the first
year.

   Concentrations of P, Cd, Ni, and Zn in wheat grain, and P, Ca, Mg, Cd,
Ni and Zn in wheat stover were increased by sludge applications (Table 26).
As was the case with rye, Cd contents of wheat tissues were enhanced by
sludge applications, but did not accumulate correspondingly higher concen-
trations of the metal with higher amounts of applied sludge-borne Cd.  Leaves
of wheat and rye (collected as heads were emerging from the boot) grown on
sludge-amended spoil had higher concentrations of Mg, Mn,  Cd, and Zn as com-
pared to those collected from control plots (Table 27).   Sludge applications
also increased concentrations of P and Ni in the leaves of rye and N in
leaves of wheat.

   After rye and wheat were harvested, in the first year  of the study, a
forage sorghum (Sorghum vulgare) was seeded in the split-plots.  Two months
later, above ground whole plant samples of sorghum were harvested and ana-
lyzed for concentrations of the several elements listed in Table 28.   For
sorghum following rye, concentrations of P, Mg, Mn,  Cd, Cu, Ni, and Zn were
increased in whole plants.  Concentrations of all of these elements except
Mg and Mn had been significantly increased in spoil by sludge additions.
Although concentrations of Ca, Fe, Cr, and Pb were Increased  in the surface  layer
(0-15 cm) of spoil their uptake by sorghum plants was not  increased.   Nickel
and Zn were the elements whose concentrations were most consistently increased
in tissues of all grass species by sludge applications.   In agreement with
data from corn, rye, wheat, and sorghum,  concentrations of elements accumu-
lated by grasses generally increased with higher sludge applications,  except
for Cd.  Concentrations of Cd in the several kinds of plant tissues generally
did not differ between the two highest sludge loading rates.   All sludge
loading rates resulted in about the same enhancement of Cd contents,  when
compared to Cd levels in plants from control plots.
                                      136

-------
Table  24.  Concentrations of selected elements in corn grain from plots with and without sludge.   Corn
           was planted with a no-till planter in dead wheat and rye.
Planted in Dead Rye
Planted in Dead Wheat
Sludge Application Rates
..I- /V,-.
Element
Year
0
224
448
896
LSD 0
224
448
896
LSD

Mg
Ca
P
N
Zn
Fe
Mn
1979
1980
1979
1980
1979
1980
1979
1980
1979
1980
1979
1980
1979
1980
0.12
0.12
0.007
0.002
0.28
0.30
1.41
1.77
27.5
25.0
19.9
20.4
8.54
7.69
0.14
0.16
0.005
0.007
0.31
0.37
1.64
1.79
33.3
37.6
21.9
31.2
7.37
6.96
0.14
0.16
0.005
0.004
0.36
0.36
1.66
1.87
36.9
43.5
22.6
33.2
7.46
7.40
0.14
0.16
0.006
0.006
0.36
0.39
1.73
2.02
42.2
52.7
22.2
27.9
B.91
10.5
0.01** 0.13
n.s. 0.15
n.s.
n.s.
0.03**
n.s.
n.s.
n.s.
-mg/kg (dry
7.86*
19.4**
n.s.
n.s.
1.18*
2.38*
0.006
0.004
0.26
0.34
1.52
1.65
weight)-
27.9
29.3
24.9
27.6
7.91
8.34
0.13
0.14
0.007
0.005
0.31
0.34
1.72
1.85
36.8
34.0
32.5
25.2
7.06
6.96
0.15
0.15
0.005
0.006
0.36
0.3fi
1.72
2.29
43.9
46.1
31.7
25.7
7.73
7.54
0.16
0.16
0.004
0.004
0.39
0.38
1.84
2.22
48.8
49.1
34.9
27.8
10.2
10.2
0.02*
n.s.
n.s.
n.s.
0.04**
n.s.
n.s.
n.s.
7.32**
14.5*
4 . 98*
n.s.
1.57**
n.s.
*, ** = sginificantly different at P£0.05 and P<0.01,  respectively.
n.s.  = no significant differences.

-------
U)
00
       Table  24.  Concentrations  of  selected  elements  in  corn  grain  from  plots with and without  sludge.  Corn

       continued  was  planted  with a no-till  planter in dead wheat and  rye.



Planted
in Read
Rye
Planted in Dead Wheat
Sludge Application Rates
Element
Ni
Cr
Pb
Cd
Cu
Year
1979
1980
1979
1980
1979
1980
1979
1980
1979
1980

0

0.63
<0.62
0.423
<0.125
<0.62
<0.62
<0.062
0.071
2.80
1.80

224

0.83
0.77
1.06
<0.125
<0.62
0.696
0.229
0.346
2.67
2.50

448

1.39
1.32
0.372
<0.125
<0.62
<0.62
0.359
0.613
2.50
2.16

896

3.43
2.51
0.471
<0.125
<0.62
<0.62
0.382
0.488
2.75
?.03
m*.
/Un
LSI) 0
kg (dry
1.38**
0.98**
U.S.
n. s.
11. S.
n. s.
0.232**
0.261*
n.s.
n. s.

<0.62
0.66
<0.125
<0.125
<0.62
0.<»4
<0.062
<0.062
2.78
1.93

224

0.62
0.7')
<0.125
<0.125
<0 . 62
<0.62
0.254
0.266
2.87
2.46

448

1.48
1.40
<0.125
<0.125
<0.62
<0.62
0.352
0.524
2.74
2.23

896

3.07
2.53
<0.125
<0.125
<0.62
0.68
0.344
0.399
2.80
2.21

LSD

1 .04**
0.80**
n.s.
n.s.
n.s.
n.s.
0.219**
0.308* ;
n.s.
n.s.
       *,  **  =  significantly different  at  P<0.05 and P^O.Ol,  respectively.

       n.s. = no  significant differences.

-------
      Table 25.   Concentrations of selected  elements  in rye grain and stover from plots with and without sludge.
VO
Rye Grain
Rye Stover
Sludge Application Rates

Element
Mg
P
N
Zn
Fe
Mn
Ca
Year
1979
1980
1979
1980
1979
1980
1979
1980
1979
1980
1979
1980
1979
1980
0

0.119
0.142
0.292
0.406
2.08
1.96
37.7
39.7
41.5
63.6
10.8
26.3
439
437
224

0.125
0.153
0.327
0.472
2.28
2.30
57.7
55.8
42.0
53.1
8.60
18.0
458
456
448

0.148
0.170
0.410
0.469
2.64
2.37
75.7
55.0
47.6
85.7
10.8
42.7
448
504
896

0.169
0.164
0.431
0.464
2.62
2.44
83.4
63.7
47.9
42.9
13.7
34.2
412
403
LSD 0
Of / J __-.
*-i«i.i^
0.020** 0.135
0.017* 0.097
0.053**
0.046*
0.42**
n.s.
-mg/kg (dry
15.3**
12.5*
5.36*
n.s.
n.s.
n.s.
n.s.
n. s.
0.040
0.070
0.54
0.44
weight)-
13.2
21.5
133
59.5
14.6
13.5
3130
1880
224

0.212
0.158
0.053
0.162
0.768
0.80
40.5
43.6
149
55.3
10.2
11.4
4840
2880
448

0.328
0.202
0.143
0.170
1 .03
0.88
94.1
71.1
136
76.7
16.0
42.0
6290
3020
896

0.485
0.296
0.229
0.226
1.11
1.01
122
106
108
69.8
35.8
25.6
5970
3040
LSD

0.112**
0.060**
0.108**
0.069**
0.202**
0.278**
60.5**
35.5**
n.s.
n.s.
11.9**
n.s.
934**
639*

-------
Table  25.  Concentrations of selected elements in rye grain and stover from plots with and without  sludge.
continued
Rye Grain
Rye Stover
Sludge Application Rates
Element
Ni
Cr
Pb
Cd
Cu
Year
1979
1980
1979
1980
1979
1980
1979
1980
1979
1980

0

<0.62
<0.62
0.178
0.863
<0.62
2.63
0.065
<0.062
6.45
5.89

224

<0.62
<0. 62
0.140
0.604
<0.62
2.63
0.252
0.329
7.18
7.23

448

2.23
1.56
0.182
1.10
<0.62
1.96
0.532
0.387
8.40
7.96

896

6.96
2.53
0.249
1.25
<0.62
3.73
0.426
0.389
9.20
7.99
ml-/
i.«
LSD 0
— mg/kg (dry
4.10**
1.11**
n. s.
n. s.
n. s.
n. s.
0.121**
0.176**
1.26**
1.29**
5 we lyht)-
<0.62
<0.62
0.611
1.74
0.862
10.2
0.069
0.124
3.55
3.16

224

<0.62
<0.62
0.764
2.13
<0.62
10.2
0.847
1.20
4.80
4.57

448

1.61
1.15
0.874
2.30
<0.62
10.3
2.03
2.11
7.34
5.73

896

4.92
2.58
0.544
2.25
<0.62
10.5
2.20
2.32
10.2
6.93

LSD

3.35**
1.20**
11 . S.
n. s.
n.s.
n.s.
1.34**
0.92**
4.31**
1.31**
 * =  significant  at  P<0.05
 **   =  significant at  P£ 0.01
 n.s. = not  significantly different

-------
Table 26.  Concentrations of selected elements In wheat grain and stover from plots with and
           without sludge.
Wheat Grain
Wheat
Stover


Sludge Application Rates
Element
Mg
P
N
Zn
Fe
Mn
Ca

Year 0 224

1979 0.195 0.220
iqan 	 	
1979 0.330 0.381
i Qftn — — __-.______.
1979 2.37 2.45
juan — — — .

1979 52.0 67.0
IQRf) - 	 - 	 -
1979 38.0 59.6
1QRO -
1979 48.7 27.4
IQQO 	 _
1979 660 452
iqart 	 	 .


448

0.184
0.393
-_— —_n^_ — -
2.63
	 „ J

68.3
— _ r»H— -
53.7
- - nA
33.7
.——_—nrl —
480
. 	 _nrl— - -


896 LSD 0

0.230 n.s. 0.196
0171
0.535 0.123* 0.040
01 7fl
2.82 n.s. 0.489
OQ 1 1

— — mg/kg (dry weight)-
84.3 19.0* 28.8
	 _ __ __ _ _ _ A7 1
53.7 n.s. 670
	 _ IAC
40.1 n.s. 34.1
— ____-. _ A 7 7
461 n.s. 2390
. ____ _______ lAin


224

0.242
0.210
0.059
0.357
0.815
1.22
55.6
89.6
459
149
23.1
24.5
3040
2560

448

0.290
0.249
0.056
0.318
0.665
1.25
60.7
92.2
238
123
18.9
29.9
3270
2940

896

0.559
0.310
0.122
0.429
0.784
1.40
108
126
184
160
30.4
44.4
4770
2500

LSD

0.267**
0.066**
0.054**
0.165*
n. s.
U.S.
45.2,**
40.2*
n.s.
n. s.
n. s.
14.4*
1490*
n. s.
n.s. = no significant differences.
*, ** = significant difference at F<0.05 and P<0.01, respectively.
nd = no data

-------
Table 26.  Concentrations  of  selected  elements  in wheat  grain and  stover from plots with and
continued  without  sludge.
Wheat Grain
Wheat Stover
Sludge Application Rates


Element Year 0 224

NL

Cr

Pb

Cd

Cu



1979 0.717 2.15
iqflri 	 	 	 	
1979 1.44 2.48
iqnn 	 	 	
1979 <0.62 <0.62
ioan 	 	 	
1979 0.248 1.08
loon 	 	 	 	

1979 4.18 5.31
iqon _


448


2.96
— —rfcrl— — —
1.07
_ — — nsl___
<0.62
— — — nrl — —
1.08
_____,-l ___

4.47
	 	 „,! 	 	


896 LSD 0
/i / j -i i »- \

8.25 3.22** 1.41
— — — — -.— — — _ .-_ n f\9
0.120 n.s. 2.42
1/iQ
<0.62 n.s. <0.62
OQ?
1.50 0.878** 0.555


4.92 n.s. 3.29
5n&


224


1.11
1 9"7
2.62
2ni
0.737
Im
1.95
3.94
5.28
7.57

448


1.23
11 1
2.34
17ft
0.715
O/i ft
2.15
4.18
5.55
6.01

896


4.33
•i to
1.85
299
0.826
079
2.76
4.50
7.24
7.52

LSD


1.60**

n. s.
01^*
n.s .

1.35*
2.79**
n.s.
n..s.
n.s. = no  significant differences.
*,  ** = significant difference  at P^O.05  and  P_<0.01,  respectively.
nd  = no data

-------
   Concentrations of selected chemical elements in eight species of grasses
grown in control and sludge-treated plots are shown in Table 29.  Stands
of big bluestern were never adequately established on spoil materials.   Samples
of other species were collected from the first cutting on June 20.   Appli-
cations of sludge significantly increased concentrations of the following
elements in various species:   N in redtop and orchard grass; P in brome,
orchard, western wheat,  and timothy; Mn in perennial rye and tall fescue;
Cd in redtop,  orchard, and western wheat; Cu in orchard, western wheat,
reed canary; Ni in brome,  orchard, western wheat, reed canary; perennial
rye, timothy,  and tall fescue; and Zn in all species except redtop. A trend
for all elements except Ca, Fe, Cr, and Pb to increase in grasses was  exhib-
ited, but analytical results  were too variable for these to be statistically
significant.  Nickel and Zn were the elements whose concentrations were most
consistently increased in tissues of all grass species by sludge applica-
tions.  In agreement with data from corn, rye, wheat, and sorghum,  concen-
trations of elements accumulated by grasses generally increased with higher
sludge applications, except for Cd.  Concentrations of Cd in the several
kinds of plant tissues generally did not differ between the two highest
sludge loading rates.

Summary and Conclusions

   The incorporation of sludge into the surface 0 to 18 cm of spoil ma-
terials produced a mixture that was about 8, 16, and 32 percent sludge for
the three respective loading rates of 224, 448, and 896 mt/ha.  Thus,  the
effect on physical properties were two fold.  Physical properties of sludge
itself were reflected in proportion to loading rates, and, secondly, some
improvement in physical properties may have occurred as a result of stimu-
lated microbial activity.   However, because very little organic carbon was
lost during the ten months that elapsed between incorporation of sludge and
measurement of aggregate stability and available moisture holding capacity,
microbes probably played only a minor role.

   Because moisture contents  at 1/3 bar increased more rapidly than at 15
bar of tension, available water holding capacity increased'from 14.8%  in
spoil without sludge to 21.1% in spoil amended with 896 mt/ha of sludge.
This increase in available moisture may have offset some of the deleterious
effects on the growth of crops that were expected as a result of higher
soluble salt contents in sludge-amended spoil.

   The electrical conductivity (25 C) of 6.6 m mho/cm for saturated ex-
tracts of maximum sludge-amended spoil was in the range where a 25 to  50%
reduction in corn yields was  expected (EPA, Water Quality Criteria, 1972).
Thus, at the highest sludge loading rate the increase in potential avail-
able water holding capacity was small in comparison to the increased osmotic
pressure of soil solution.  Corn yields on plots treated with 224 mt/ha of
sludge were about 50% higher than those on plots treated with 896 mt/ha and
it appears that this reduction was due to high concentrations of soluble
salts.  Both rye and wheat are more tolerant of high salt conditions than
is corn and the results of this study show that if yields of small  grains
were affected by soluble salts, it was a rather nominal effect.  However, it
was probably a major factor affecting the establishment of some of  the grasses.

                                      143

-------
Table  27.  Concentrations of selected elements In leaves of rye and wheat from plots with and
           without sludge.




Rye




Wheat


Sludge Application Rates

Element
Mg
Ca
P
N
Zn
Fe
Mn
Ni
Year
1980
1980
1980
1980
1980
1980
1980
1980


0
0
0
2
13
333
32
<0
0 224

.171 0.348
.589 0.780
.180 0.275
.73 3.29
.4 21.9
221
.0 26.1
.62 <0.62
448

0.373
0.768
0.290
3.22
30.4
374
33.7
0.786
896

0.512
0.764
0.348
3.51
39.9
284
61.5
1.05
LSD 0
ty / j
0.116**
n.s.
0.090**
• i - \
0.276
0.245
0.370
n.s. 4.14
— mg/kg (dry weight)-
11.7** 19.7
n.s.
21.2**
0.50*
144
60.7
<0.62
224

0.384
0.377
0.507
4.73
27.0
126
36.2
<0.62
448

0.456
0.437
0.522
4.95
28.2
120
51.4
<0.62
896

0.565
0.534
0.554
4.82
35.7
112
87.8
<0.62
LSD

0.174**
n.s.
U.S.
0.17**
10.1**
n.s.
29.7*
n.s.
n.s. = no significant differences.
*, ** = significant difference at P<0.05 and P<0.01, respectively.

-------
Table 27.  Concentrations  of selected elements in leaves of rye and wheat  from plots with and
continued  without  sludge.




Rye




Wheat


Sludge Application Rates

Element

Cr
Pb
Cd
Cu

Year

1980
1980
1980
1980


0


1.24
1.23
0.079
7.51

224


1.11
1.67
0.407
10.1

448


1.73
2.82
0.696
16.5

396


2.30
2.74
0.924
13.6

LSD
-mg/kg (dry
n.s.
n.s.
0.402**
5.21*

0
• i *. \

0.850
<0.62
0.312
9.11

224


0.753
<0.62
0.668
10.8

448


0.850
0.86
1.04
10.4


896 LSD


0.
<0.
1.
10.


931 n.s.
62 n.s.
34 n.s.
3 n.s.
n.s. = no significant differnences.
*, ** = significant difference at P_<0.05 and PfO.Ol, respectively.

-------
Table  28.  Concentrations of selected elements in sorghum whole plant from plots with and without
           sludge.  Sorghum was planted in dead wheat and rye in July of 1979.
Planted in Dead Rye
Planted in Dead Wheat
Sludge Application Rates
Element
Mg
Ca
P
N
Zn
Fe
Mn
Ni
Cr
Pb
Cd
Cu

0

0.312
0.322
0.217
1.53
45
143
30
0.933
0.614
<0.62
0.368
7.47

224

(J.335
0.304
0.184
2.02
55
110
23
0.571
0.402
<0.62
3.87
9.31

448

0.469
0.285
0.179
2.00
69
125
35
0.994
0.615
<0.62
3.10
10.6

896

0.511
0.340
0.180
1.82
77
109
51
1.81
0.440
<0.62
4.83
10.3
.
- /K™
LSD 0
Zf j_.
0.152*
n.s.
0.027*
n.s.
mg/kg (e
20**
n.s.
17**
0.647*
n.s.
3.14**
1.74**

0.313
0.291
0.213
1.86
Iry weight)
49
160
29
0.998
0.455
<0.62
0.571
9.30

224

0.398
0.340
0.223
2.17
66
118
28
1.09
0.341
<0.62
6.41
9.03

448

0.554
0.369
0.258
2.51
110
122
41
2.15
0.645
<0.62
10.7
11.7


896

0.
0.
0.
2.
137
137
88
3.
0.
9.
11.

688
322
251
29



78
660
62
93
9

LSD

0.129**
0.048**
n.s.
0.40**
46*
n.s.
26**
1.22**
n. s.
6.20*
n.s.
n.s. = no significant differences.
*, ** = significant difference at P^O.05 and P<0.01, respectively.

-------
Table 29 .   Concentrations of selected elements in various grasses  grown
           in 1980 on plots with and without sludge.
Sludge Application Rates
Analyte


Mg
Ca
P
N
0



0.177
0.227
0.203
0.973

224



0.224
0.246
0.236
1.76

448
Redtop
e/ f A U ^\

0.244
0.276
0.230
1.52

896



0.242
0.209
0.218
1.93

LSD



n.s.
n.s.
n. s.
0.591*
	 mg/kg (dry weight) 	
Zn
Fe
Mn
Ni
Cr
Pb
Cd
Cu
20.5
52.1
61.9
0.996
0.750
<0.62
<0.062
4.55
44.5
64.4
59.3
2.74
0.876
<0.62
0.296
6.61
61.3
63.7
75.5
5.17
0.199
<0.62
0.954
7.33
59.0
51.8
86.6
7.90
0.440
<0.62
0.684
7.33
n.s.
n.s.
n.s.
n.s.
0.443*
	
0.618*
n.s.



Brome


	 % (dry weight) 	
Mg
Ca
P
N

Zn
Fe
Mn
Ni
Cr
Pb
Cd
Cu
0.120
0.185
0.135
1.19
	
15.3
140
45.6
<0.62
0.893
<0.62
0.417
6.14
0.196
0.270
0.194
1.26
	 mg/kg
33.6 .
71.7
23.4
0.837
0.692
<0.62
1.28
6.76
0.196
0.234
0.240
1.65
0.290
0.270
0.291
1.62
0.082*
0.065**
0. 108**
n.s.
(dry weight) 	
49.0
63.4
28.5
1.91
0.884
<0.62
1.69
7.57
61.7
99.8
61.0
3.50
1.62
<0.62
2.21
10.3
24.5**
n.s.
20.8*
1.64**
n.s.
	
n.s.
n.s.
*,** - Significantly different at P  <_ 0.05  and P _< 0.01,  respectively
n.s. - No significant differences

                              147

-------
Table 29 .
continued
Concentrations of selected elements in various grasses gr<
in 1980 on plots with and without sludge.
Sludge Application
Rates

me /U-,
Ar.alyte
0
224 448
896
LSD
Orchard
	 % (dry weight) 	
Mg
Ca
P
N

Zn
Fe
Mn
Ni
Cr
Pb
Cd
Cu
0.293
0.307
0.203
1.07
	
19.5
94.5
106
1.50
0.392
<0.62
0.064
4.42
0.297
0.264
0.267
2.29
	 mg/kg
36.2
75.0
39.6
2.17
0.428
<0.62
1.08
9.18
0.316
0.270
0.311
2.35
0.358
0.228
0.365
2.56
n.s.
n.s.
0.086*
0.98**
(dry weight) 	
50.6
86.3
52.5
5.07
0.646
<0.62
1.86
11.8
66.6
78.8
122
12.2
0.598
<0.62
1.87
13.2
27.6**
n. s.
52.8**
6.03**
n.s.
	
1.00*
4.39*
Western Wheat

Mg
Ca
P
N

Zn
Fe
Mn
Ni
Cr
Pb
Cd
Cu

0.131
0.166
0.141
1.32


18.0
57.9
30.2
<0.62
1.16
<0.62
<0.062
3.89
_——__— — — v (,
————«—— ^ ^{
0.154
0.209
0.187
1.55
.

33.6
75.6
22.6
<0.62
0.972
<0.62
0.815
4.92
, . , .
0.158
0.185
0.191
1.53
(dry weigh
40.0
65.2
22.6
1.41
1.08
<0.62
1.07
5.41

0.195
0.167
0.222
1.60
t.\
it; 	
45.6
85.1
33.6
2.45
1.71
<0.62
0.892
6.24

0.033*
n.s.
0.040*
n.s.


20.0**
n.s.
7.8*
1.47**
n.s.
___
0.720**
1.12*
*,** - Significantly different at P <_ 0.05 and P _< 0.01, respectively
n.s. - No significant differences
                                  148

-------
Table '29. Concentrations of selected elements in various grasses gr
continued in 1980 on plots with and without sludge.
Sludge Apolicatio'n Rates
Analyte

Mg
Ca
P
N

Zn
Fe
Mn
Ni
Cr
Pb
Cd
Cu

0


0.244
0.261
0.240
1.64


34.6
66.5
59.8
1.74
0.991
<0.62
<0.062
6.18

224
Ree
_ °j i

0.275
0.258
0.349
2.29

	 mg/ Kg
95.9
72.5
56.8
4.17
1.08
<0.62
0.539
9.35

448
d ' s Canary
dry weight
0.300
0.258
0.383
2.24

896
r
,\
- ) 	
0.358
0.228
U.399
2.23

LSD


n.s.
n.s.
n.s.
n.s.
(dry weight) 	
131
91.2
86.8
8.98
1.39
<0.62
1.09
11.3
125
79.8
96.0
11.5
1.22
<0.62
0.773
12.4
51.8*
n.s.
n.s.
5.08**
n.s.
	
n.s.
4.50*
                                Perennial Rye

                            	% (dry weight)-
Mg
Ca
P
N

Zn
Fe
Mn
Ni
Cr
Pb
Cd
Cu
0.216
0.252
0.205
1.02


21.8
87.2
41.4
1.09
0.964
0.66
<0.062
3.83
0.282
0.316
0.260
1.68
.
•— — — mg/Kg
43.9
70.0
37.9
3.10
0.904
<0.62
0.129
6.10
0.335
0.436
0.357
2.27
(dry wei|
73.9
90.4
61.9
9.24
0.915
<0.62
0.887
10.2
0.403
0.454
0.411
1.73
»Vit- \ _________
gntj 	
116
190.?
144
22.8
2.64
4.84
2.51
16.5
n.s.
n.s.
n.s.
n.s.


49.3*
n.s.
42.3**
9.79**
n.s.
n.s.
n.s.
n.s.
*f**
       Significantly different at P _< 0.05 and P _< 0.01, respectively
n.s. - No significant differences
                                   149

-------
Table 29 • Concentrations of selected elements in various grasses gn
continued in 1980 on plots with and without sludge.
Sludge Application Rates

Analyte


Mg
Ca
P
N
	
0

	
0.120
0.163
0.214
1.24
	
224



0.163
0.187
0.247
1.36
	 mt /ha 	
448
Timothy
	
896

	
LSD

—7. (dry weight) 	
0.175
0.198
0.267
1.38
0.227
0.175
0.272
1.18
0.061**
n.s.
0.032*
n.s.
	 mg/kg (dry weight) 	
Zn
Fe
Mn
Ni
Cr
Pb
Cd
Cu


Mg
Ca
P
N
26.3
107
31.7
0.816
1.10
<0.62
<0.062
7.58



0.214
0.252
0.191
1.17
46.4
67.6
30.9
2.29
1.05
<0.62
<0.062
6.89



0.291
0.299
0.233
2.23
75.6
59.8
39.3
4.25
1.17
<0.62
0.454
8.31
Tall Fescue


0.321
0.298
0.267
2.15
82.2
65.6
51.8
9.94
1.40
<0.62
0.485
10.1



0.339
0.263
0.278
1.92
39.1*
n.s.
n.s.
6.43**
n.s.
	
n.s.
n.s.



n.s.
n.s.
n.s.
n.s.
	 mg/kg (dry weight) 	
Zn
Fe
Mn
Ni
Cr
Pb
Cd
Cu
* t** _
n.s. -
20.8
76.0
31.7
1.10
1.03
<0.62
0.318
4.40
Significantly
No significant
28.0
80.0
25.9
1.82
0.414
<0.62
0.674
5.53
42.4
91.5
36.8
4.84
1.00
0.665
1.13
6.69
different at P <_ 0.05 and
46.8
88.4
61.8
7.17
0.784
<0.62
1.28
6.70
P < 0.01,
16.1*
n.s.
18.4**
2.42**
n.s.
n.s.
n.s.
n.s.
respectivel;
: differences
150

-------
Western wheatgrass has high sale tolerance and while perennial ryegrass and
call fescue have only medium tolerance, they are more tolerant than the other
grass species used in this study.  Soluble salts appear to be the major factor
affecting crop growth and survival of grasses at the seedling stage on
spoil amended with sludge at loading rates which exceed 224 mt/ha.

   For elements that had increased concentrations in spoils as a result of
sludge applications and which were accumulated by corn plants by uptake and
translocation into leaves and grain, concentration ratios (CR) were calcu-
lated and are presented in Table 30.  Indigenous concentration ratios were
obtained by dividing the concentrations of a particular element in leaves
or grain from control plots by total concentrations of that element in spoil
materials that were not treated with sludge.  Amended concentration ratios
were calculated by subtracting concentrations in leaves or grain from con-
trol plots from those in similar tissues from sludge-amended plots and divid-
ing by the remainder obtained by subtracting indigenous concentrations in
spoil from concentrations in sludge-amended spoil.  These concentration
ratios are similar to those presented by Cataldo and Wildung (1978), except
that they added a single concentration of each element (2.5 nig/kg) to the soil.

   The CRs that could be calculated for corn leaves (Table 30) show that,
except for Mi, the constituents of sludge were either not as available for
uptake as indigenous elements or their uptake was limited by metabolically
regulated processes.

   In soil amended with soluble salts of metals, Cataldo and Wildung (1978)
found CRs increased for As, Co, Cr, Mn, Mb, Ni, Pb, Sb, and Zn.  In this
study Mn was not increased in spoil by sludge applications because concen-
trations in sludge were no higher than those in untreated spoil materials.
Previous analysis of sludge from the wastewater treatment plant showed that
As, Co, and Mo concentrations were too low to increase total concentrations
in normal soils (Hinesly and Sosewitz, 1969) and this has been borne out by
determining concentrations in soils treated with annual sludge applications
beginning in 1967 (Hinesly and Hansen, 1979).  Concentration ratios could
not be calculated for Cr and Pb because they were not increased in corn tis-
sues, although levels of these two metals were markedly increased in sludge-
amended spoil.

   Concentrations of Ca in corn grain were not affected by sludge treat-
ments as they were in leaves, so CRs for this element could not be calcu-
lated for grain (Table 30).  Iron concentrations in leaves were not affected
by sludge treatments, but were in grain from corn grown in wheat mulch.
Since Zn concentrations were higher in grain from wheat mulch as compared to
rye mulch plots, CRs from Zn in grain from the two different plots were cal-
culated separately.  As was the case for leaves, all CRs for grain produced
on sludge-amended spoil were lower than those for grain from control plots,
except for Ni.  Concentration ratios for Ni tended to increase with in-
creased sludge loading rates.
                                      151

-------
Table  30.  Corn Leaf and Grain (1979) Concentration Ratios for Comparing
           the Uptake of Indigenous Elements to Those Added as Constitu-
           ents of Sludge.
A.



ELEMENT
N
P
Ca
Mg
Zn
Cd
Ni
CORN



Indigenous CR
27.70
2.93
0.96
0.48
0.60
0.27
0.007
LEAF CONCENTRATION RATIOS

Sludge


224
1.57
0.07
0.13
-0.38
0.07
0.06
0
Amended CR
Application


448
1.14
0.04
0.12
-0.01
0.10
0.12
0

Rates


896
0.88
0.03
0.02
0.29
0.10
0.10
0.009

B.



ELEMENT
N
P
Mg
Fe (W)
Zn (R)
Zn 00
Cd
Ni
CORN



Indigenous CR
14.30
3.43
6.62 x 10~4
0.324
0.329
0.043
0.007
LEAF CONCENTRATION RATIOS

Sludge


224
1.64
0.05
0.28 t
10.6 x 10~
0.009
0.014
0.005
0.009
Amended CR
Application
___m<- /Vi9_____

448
0.77
0.04
0.12
* 2.66 x 10
0.006
0.011
0.003
0.012

Rates


896
0.71
0.03
.4 °'07 -4
2.22 x 10
0.006
0.009
0.002
0.018
(R) = Corn on rye.
(W) = Corn on wheat.
                                     152

-------
   Because the pH was reduced from 7.5 to 6.0 on spoil amended with 896
int/ha of sludge the decrease in concentration ratios with higher sludge load-
ing rates is contrary to expectations.  Also, since the highest corn yields
were obtained with 224 mt/ha of sludge it is contrary to expectations that
CR's decreased with higher loading rates that resulted in lower yields.
Except perhaps for Ni, there is no evidence that metal concentrations in corn
tissues were increased as a result of reduction in growth.

   The corn hybrid used in this study was the same as that used in the pre-
vious study discussed in this report where sludge from the same treatment
plant was applied each year at maximum annual loading rates that were about
50 mt/ha.  In some years, this hybrid accumulated Cd concentrations in grain
of around 1 mg/kg.  Therefore, it is unlikely that the accumulation of Cd
and other elements in corn tissues from this high-rate study was limited by
metabolic controls.  Rather, it appears that with these exceedingly high
sludge loading rates, some of the elements were less available for uptake.
At such high loading rates the availability of some metals may be controlled
to a much greater extent by the properties of sludge itself than those of
the weathered geological materials.  In the presence of excessive levels of
oxidizable sludge organic matter, sparingly soluble sulfide forms of some
metals may be rather stable and thus, availability of metals for uptake by
plants may be maintained at low levels until the organic matter has been de-
composed.  However, it seems unlikely that they will become more available
in time because results from other studies discussed in this report showed
that Zn and Cd uptake decreased after sludge-applications were terminated.
In view of the findings reported by others (Cunningham et al.,  1975), it
seems more likely that the high amounts of Fe and P supplied as constituents
of sludge limited the availability of some of the metals by forming spar-
ingly soluble complexes.

   Based on the results of feeding studies (Hinesly and Hansen, 1979, Hinesly
et al., 1981), there is no indication that feeding the grains and forages
produced on strip-mine spoil amended with high rates of sewage sludge would
present a potential health hazard to animals.  Feeding the materials to ani-
mals would present a nominal impact on food-chains, if any at all.  Consider-
ing the relatively low enhancement of Cd and high enhancement of several es-
sential trace elements, especially Zn and Cu, feed materials produced on
sludge-amended spoil may actually be a higher quality feed than that produced
with inorganic fertilization.

   Further work is needed to determine the rate and extent of the migration
of trace elements to deeper depths in calcareous spoil amended with high
loading rates of sludge.  Also, further work is needed to determine changes
in trace element availability to plants with time under these conditions.


DIFFERENTIAL ACCUMULATIONS OF CADMIUM AND ZINC BY CORN (Zea Mays L.) HYBRIDS
GROWN ON SOIL AMENDED WITH SEWAGE SLUDGE

Introduction

   In a prior study twenty corn inbreds, commonly used as parents of hybrids


                                      153

-------
adapted to the cornbelc region, were screened according to their capacities
to accumulate Zn and Cd in leaves and grain when grown on Blount silt loam
(Aerie ochraqualf, fine, illitic, mesic) with and without amendments with a
metalliferous sewage sludge (Hinesly et al., 1978).  Where maximum amounts
of sludge had been applied annually for seven years, the soil surface (0-15
cm) contained 454 and 21 mg/kg of Zn and Cd, respectively, compared to 68
and 0.3 mg/kg in control plots treated with commercial fertilizers.  Con-
centrations of Zn in leaves from different inbr&ds grown on maximum-sludge-
treated plots ranged from 62 to 282 mg/kg and concentrations in grain from
three inbreds (H99,  Oh545, and R805) were not significantly increased by
sludge-borne applications of Zn and Cd.  Such extreme differences in capaci-
ties to accumulate Zn and Cd as were found in this small sample of inbreds
raised the question of whether or not the differences in Zn and Cd accumu-
lations were genetically predisposed, and, if so, how they were inherited.

   Before experiments could be designed to determine the nature of the
genetic component controlling the accumulation of transition metals by maize,
additional information was required.  Results from greenhouse and field
studies are discussed here providing information about 1) factors external
to roots of one inbred affecting the accumulation of Cd by an adjacent dif-
ferent inbred; 2) maternal factors influencing the accumulation of Zn and Cd
by hybrids; and 3) the constancy of predicted differential capacities of
hybrids to accumulate Cd under field conditions.

Materials and Methods

Root Interaction Study—

   Ipava silt loam (aquic arguidoll, fine, montmorillonite, mesic, pH 5.5)
soil was collected from the end of a field that had received 202 mt/ha (dry
weight equivalent) of liquid digested sewage sludge in four years.  The soil
was air-dried, ground, and mixed prior to subdividing into three-kg portions
which were placed in plastic pots (20.3 cm diameter).  Subsamples were taken
for Cd analysis.  Water was added to the pots until the soil was completely
saturated, after which it was allowed to drain for three days.  Four seeds
of a single cross selected to take up high (H98 X B37) or low (B73 X R805)
amounts of Cd were planted in each half of the surface area of a pot, until
all possible pairs of different or the same inbreds (10) had been made.  All
pairs were replicated six times.  All pots (12 rows) were rotated (2 rows to
the north) on the greenhouse bench once each week and water was added as neces-
sary to maintain soil moisture near field capacity.  Ten days after planting,
the number of plants was reduced to two.  Thirty-four days after planting,
one-half of the pots (three replications) were randomly selected from which
whole plants were harvested.  Three leaves adjacent to ear nodes, were har-
vested 89 days after planting from each plant growing in the remaining 30
pots.  All samples were prepared and analyzed for Cd concentrations accord-
ing to methods described below for the field study.

Maternal Influence Study—

   Five inbreds were selected from the screening study that had shown a low
to high propensity for accumulating Cd and another five were selected for
their range in capacities to take up Zn.  Using the selected inbreds, re-


                                      154

-------
ciprocal crosses were made between chose that accumulated high and low
amounts of Cd.  Two of the hybrids were crosses of related inbreds (B37 X
B73 and H98 X Oh545) and two were unrelated (B37 X R805 and H98 X B73) crosses.
Analogous reciprocal crosses were made between related and unrelated inbreds
that accumulated high and low amounts of Zn.  Related crosses were B37 X B14A
and B37 X R802A and unrelated crosses were A619''X H98 and A619 X 314A.  These
crosses were mada in the University of Illinois South Farm corn breeding
nursery.  Ten seeds of each inbred and single-cross were planted in pots of
sludge-amended Ipava silt loam soil as described above for the root inter-
action study.  Each inbred and hybrid planting was replicated five and six
times, respectively.  Ten days after planting the number of plants per pot
was reduced to four.  Leaves from these four plants were harvested at six
weeks from the date of planting.  Daily maintenance was identical to that
previously described for the root interaction study.  All soil and plant
tissue samples were analyzed for Cd and Zn concentrations by methods de-
scribed below for the field study.

Field Study—

   Single-crosses Mol7 X H98 and Oh545 X B73 were selected as high and low
Cd accumulators, respectively, and planted each year in split-plots of Blount
silt loam (Aerie ochraqualf, fine, illitic, mesic, pH 7.4) soil with and
without amendment with sewage sludge.  The plots measuring 3.1 x 15 m were
separated by a border area 3.1 m wide.  The plots were formerly used to study
changes in water quality and the first annual sludge application was made 9
years prior to the initiation of this study.  Three replications for each of
the three different sludge loading rates and a control (no sludge) were ran-
domized within the study area.  Liquid digested sludge from the Southwest
Wascewater Treatment Plant in Chicago, Illinois, containing 1.5 to 3.5%
suspended solids, was applied each year by furrow irrigation at approximate
plot depths of 6.4, 12.7, and 25.4 mm.  At each sludge application, control
plots were irrigated with 25.4 mm of water.  Successive applications were
made when sufficient sludge moisture had been lost by infiltration and evapo-
ration to permit additional sludge to be added to the maximum-sludge-treated
(25.4 mm) plots.  Thus,  the frequency of sludge applications varied as a re-
sult of differences in climatic conditions during the growing season.  All
plots were fertilized with a broadcast application of KC1 to supply 112 kg K/ha
prior to plowing, ridging, and planting operations.  Control plots also re-
ceived 336 kg N/ha as NH NO, and 112 kg P/ha as triple superphosphate.
                        4  J
   Seeds of either the Mol7 X H98 or Oh545 X B73 hybrid were hand-planted
on ridges of randomly selected halves of plots at rates to provide popula-
tions of 60,000 plants/ha after thinning.  When about 15% of the plants
tasseled, the leaf adjacent to the ear node was collected from 10 plants in
each split-plot, washed in distilled water, dried at 60°C, and ground to pass
a 20-mesh screen.  Subsamples of grain, hand-harvested from split-plots to
determine yields, were dried and ground in the same manner as leaf samples.
After grain harvest, 12 and 13 consecutive plants from the two center rows
were harvested and weighed for a determination of stover yields.  From the 25
plants, four plants were randomly selected, chopped into short segments with
a corn knife, dried at 60 C to determine moisture contents, and ground to
pass a 20-mesh screen.  Leaf, grain and stover samples (2g) were digested in
concentrated HNO-j at 90°C, followed by HCIO^, at 200°C taken to dryness, dis-


                                      155

-------
solved in 0.1 £ HNO. and analyzed for Ca,  Mg,  K,  Fe,  Mn,  Cd,  Cu,  Cr,  Ni,  Pb,
and Zn concentrations by atomic absorption spectrophotometry  with appropriate
background correction.  Phosphorus concentrations were determined colori-
metrically with the vanadomolybdate finish (Greweling, 1976)  and  N by a
method described by Bremner (1965).

   Soil samples were collected with stainless  steel tubes to  a depth  of 76
ca from each plot following the first spring tillage  operation and segmented
into depths of 15 cm.  After drying (60 C), crushing, and pulverizing to  pass
a 60-mesh screen, aliquots (0.5 g) were heated to 500 C for 24 hours,  digested
in concentrated HCl-HF, and dissolved in 1 If HC1  for  determining  metal con-
centrations by atomic absorption spectrophotometry.

Results

   The sludge-amended Ipava soil used for the  greenhouse studies  contained
57.4 + 2.9 mg Cd/kg and 864 + 25 mg Zn/kg.  Thus, this soil contained about
115 and 10 times more Cd and Zn, respectively, than expected  in a noncon-
taminated condition.

   Inbreds H98 and B37 compared in the root interaction study accumulated
significantly more Cd in whole plants (first harvest) and leaves  (second
harvest) than did inbreds B73 and R805, regardless of which inbred they were
paired with in pots of sludge-amended Ipava (Table 31).  Leaves collected
near the tasseling period of growth contained  significantly less  Cd than  did
young whole plants, regardless of whether the  inbreds accumulated low or  high
amounts of the metal.  When R805 was paired with  H98, it accumulated  signifi-
cantly more Cd in whole plants than when paired with  either itself or B73.
However, this was not the case for Cd concentrations  in leaves from older
plants of the same inbred pairs.

   The wide differences in capacities to take  up  Cd and Zn by the corn in-
breds used to investigate maternal effects may be seen in Table 32.  Inbred
B73 accumulated significantly less Cd in leaves than  other inbreds, except
R805.  Reciprocal crosses between high Cd accumulators (B37 and H98)  and
low Cd accumulators (B73, R805, and Oh545) contained  about the same con-
centrations of leaf-Cd.  Regardless of which inbred was used  as the female,
Cd concentrations in leaves from single-crosses were  intermediate to  those
in leaves from parent inbreds.  Single-crosses containing B73 as  one  parent
accumulated significantly less Cd in leaves than  crosses containing either
OH545 or R805.  Relatively speaking, the differences  in capacities of inbreds
to accumulate Zn were not as wide as those selected as high and low Cd ac-
cumulators.  Nevertheless, U99 accumulated significantly less and A619 sig-
nificantly more Zn in leaves than other inbreds.   Leaf-Zn concentrations
were about the same in reciprocal crosses, showing the lack of maternal ef-
fects.  Crosses containing inbred A619 had significantly more Zn  in leaves
than was found in those crosses composed of inbreds that accumulated  lesser
amounts of Zn.  Where A619 was used as one of  the parents,  leaves of  single-
crosses contained Zn levels that were intermediate to those in leaves  from
parents.  But, hybrids composed of parents that accumulated similar concen-
trations of Zn in leaves had leaf-Zn concentrations that were lower than
those of either parent.


                                      156

-------
Table 31. Comparison of Cd concentrations in whole plants  (1st harvest)  and  leaves
          (2nd harvest) of selected corn inbreds when  paired with  themselves  or  diffe-
          rent inbreds in pots of soil amended with  sewage  sludge.
Primary
Inbred
1198
D37
D73
R805
Harvest
1st
2nd
1st
2nd
1st
2nd
1st
2nd

1198
90.9
39.7
102.0
64.1
11.0
2.36
16.8
0.80

D37
86.0
25.5
100.0
58.0
9.38
1.35
15.3
1.15
Companion
1)73
Cd/kg(dry
94.7
28.0
90.7
67.2
9.67
2.21
12.6
1.85
Inbred
R805
80.0
23.5
90.8
51.6
11.4
0.93 .
12.5
1.22

LSD(KO.Ol)
n.s.S/
U.S.
n.s.
n.s.
n.s .
n.u.
3.0
n.s.
£/ Not significantly different

-------
Ui
OO
     T.-ible  J2.  Concentrations (mg/kg dry weight) of Cd and Zn in  leaves of corn inbreds and reciprocal
               dosses grown in pots of silt loam soil amended with sewage sludge.  Inbreds listed  first
               in  single-crosses were used as females.


                                               Leaf-Cd for Inbreda 
-------
   When the field study was initiated in 1978,  maximum-treated plots had al-
ready received a nine-year accumulation of 483 rat/ha of solids that con-
tained 129 kg/ha of Cd and 2.275 kg/ha of Zn (Table 33).   Other plots had
received one-fourth and one-half of these amounts.   During the first two
years of the study an additional 115 mt/ha of sludge solids were applied on
maximum-treated plots which concomitantly supplied  31 and 541 kg/ha of Cd
anc. Zn, respectively.  Sludge was also applied during the last year of the
study, but the solids remained on the surface and it is known from other
studies that their content of Cd and Zn would have  affected levels in plant
tissues to only a minor extent (Hinesly et al.,  1981).

   In Table 34, it can be seen that at the beginning of the study sludge
applications had increased Cd contents in the soil  surface (0-15 cm) from
about 0.5 mg/kg to 6, 15, and 28 rag/kg in plots irrigated with sludge rates
of 6.4, 12.7,  and 25.4 mm, respectively.  Correspondingly, soil-Zn contents
had been increased from 72 mg/kg to 184, 348, and 528 mg/kg.  Additional
sludge applications during the study period increased soil contents of Cd
and Zn, not only in the surface but to some extent  in the 15-30 cm depth.
Concentrations of these metals were not increased in soil at depths below
30 cm.  A great deal of year to year variations in  metal  concentrations in
soil were due to the small number of core samples (6 per  plot) that were
purposely kept at a minimum to preserve the integrity of  the plots for a long-
term study.  Nevertheless, the data had sufficient  precision to demonstrate
the significant long-term increases of Cd and Zn in the soil.  It is note-
worthy that recovery of Cd and Zn from the soil relative  to amounts applied
(Table 34) is low.  This phenomenon has been discussed in detail (Hinesly et
al., 1981) and requires further investigation.

   Grain and stover yields are shown in Table 35.  During the first year
hybrid Mol7 X H98 produced less grain on all sludge-treated as compared to
control plots.  Less grain was produced by Oh545 X  B73 on one-fourth- and
one-half-maximum-sludge-treated plots than on control plots.  Rainfall was
extremely low during the first growing season and,  thus,  it was expected
that yields would be markedly higher on highly fertilized and water irrigated
control plots.  Apparently, the maximum liquid sludge applications allevi-
ated the moisture stress conditions to some extent, but lesser sludge loading
rates provided little relief from the condition. During  the second year
rainfall was adequate and well distributed for high corn  yields and under
these conditions yields from Mol7 X H98 were not increased by sludge treat-
ments.  But, in this one year the highest grain yields by Oh545 X B73 were
on plots irrigated with the two higher rates of sludge.  Rainfall was below
normal in 1980, but grain yields by the two hybrids were  not significantly
affected by treatments.  Grain yields of Oh545 X B73 were significantly
higher in the second (P £0.01) and third year (P _<_  0.05)  when compared to
those produced by Mol7 X H98.

   The only difference in stover production associated with treatments oc-
curred during the last year when Mol7 X H98 produced less on sludge-treated
as compared to control plots.  During each of the three years Oh545 X B73
produced more stover (P <_ 0.05) than the hybrid selected  to take up high
amounts of Cd.
                                      159

-------
Table  33. Annual amounts of  liquid digested, sewage sludge applied  on
          maximum-created Blount  plots  and  accumulative amounts  of
          solids, Cd, and Zn constituents of sludge.
             Liquid Sludge
    Accumulative
Constituents (dry wt)
Year

1969
1970
1971
1972
1973
1974
1975
1976
1977
1978
1979
1980
Total
Depth

99
147
373
175
292
233
189
236
142
189
142
142
Avg Solids
Contents
- -Z 	
1.56
2.45 •
2.78
1.86
2.02
3.04
2.78
3.03
2.67
3.53
3.42
3.54
Solids
-mt/ha-
19.3
55.5
160.2
192.4
251.1
321.9
374.3
445.6
483.3
549.8
598.2
648.2
Cd
	 _
5.2
27.4
51.4
57.2
63.9
85.6
99.0
118.1
128.6
147.9
159.7
170.2
Za
kg/ha 	
122
434
898
1033
1257
1607
1852
2125
2275
2537
2716
2891
                                   160

-------
Table 34.  Concentrations of Cd and Zn  in Blounc silt  loam  plots, with and
          without annual sewage sludge applications.  Sludge applications
          were initiated in 1969.
                  Cadmium
                                  Zinc
                          Sludge  Irrigation Rates  (mm)
        Year
6.4   12.7  25.4  LSD
6.4  12.7   25.4
LSD
ueptn
cm


0-15







15-30









1971
1972
1973
1974
1975
1976
1977
1978
1979
1980
1973
1974
1975
1976
1977
1978
1979
1980


0.74
<0.25
0.27
0.57
<0.25
0.32
0.53
0.47
0.44
0.72
<0.25
<0.25
0.38
<0.25
<0.25
<0.25
<0.25
<0.25


2.52
1.58
3.78
3.78
3.60
6.25
6.55
7.77
7.17
8.97
<0.25
<0.25
0.52
<0.25
<0.25
<0.25
0.47
0.53


4.59
2.62
6.00
6.13
8.72
12.7
15.2
17.7
17.6
14.3
<0.25
<0.25
0.46
0.38
0.67
1.39
0.47
0.67
-tng/kg (dry wt
8.47
5.90
11.4
11.8
21.1
20.6
27.8
28.3
30.1
26.3
<0.25
<0.25
0.56
0.37
0.37
0.50
1.46
0.61
3.87**
3.49**
3.28**
1.33**
5.60**
5.75**
3.63**
6.29**
5.66**
4.87
n.s.a/
n.s.
n.s.
n.s.
n.s.
n.s.
0.75**
n.s.
. \

73
72
67
72
69
68
72
77
73
79
65
70
67
63
66
68
58
79


106
103
113
147
150
188
184
216
200
268
68
94
75
79
73
81
71
82


174
122
166
196
250
326
348
385
390
321
68
80
73
75
85
94
65
77


241
220
270
308
432
454
528
602
606
537
76
110
84
71
86
70
86
84


55**
30**
94**
56**
91**
75**
38**
125**
94**
127**
94**
127**
10*
n.s.
13**
23**
14**
n.s.
** Significantly different at P£0.01
 * Not Significantly different at P<0.05
£/ Not significantly different
                                   161

-------
 Table 35. Grain (adjusted Co 15.51 moisture) and stover (dry we.) yields
           produced by two corn hybrids on split-plots of Blount silc  loam
           with and without sewage sludge.
                Mo 17 X H98
                                               Oh545 X 373
                             Sludge Irrigation Rates (mm)
 Year  0
6.4   12.7  25.4
LSD
6.4  12.7  25.4
LSD

Grain


 1978 8.34  6.34   6.64
 1979 9.26 10.1   10.3
 1980 9.02  9.08   8.56
             7.38  1.48**
             8.70  n.a.
             7.76  n.s.
              8.62  6.02
             10.9  11.6
             10.1  10.0
       6.51  8.42
      12.7  12.9
       9.48  9.30
0.94**
1.32*
                                     Stover
1978
1979
1980
6.35
7.72
9.31
4.95
6.81
6.34
3.92
7.29
7.12
5.73
8.54
7.84
n.s.
n.s.
0.36*
6
7
9
.59
.97
.52
5.82
8.04
7.99
6.78
8.87
9.73
6.54
8.61
9.89
n.s .
n.s.
n.s.
** Significantly different at P<0.01
 * Significantly different at P£0.05
a/ Not significantly different
                                       162

-------
   Cadniium concentrations in leaves, grain,  and stover of the high (Mol7 X
H98) and low (Oh545 X B73) Cd accumulating hybrids are exhibited in Table 36.
Cadmium concentrations were increased in all tissues of both hybrids with
increasingly higher sludge loading rates, except in grain of Oh5&5 X B73
during the last year.  For all treatments and years grain from Oh545 X B73
contained less Chan detectable levels (0.062 mg/kg) of Cd, except that from
maximum-sludge-treated plots.  To statistically analyze these data for grain-
Cd concentrations,  less than detectable values were scored as one-half the
detectable limit.  Sludge applications made during the study period may have
caused successively higher concentrations of the metals in stover produced by
both Mol7 X H98 (? <. 0.01) and Oh545 X B73 (P <. 0.05).  However, in several
instances concentrations of the metals in stover from control plots also
increased each year and thus some of the increases may have been in response
to differences in climatic conditions (CAST, 1980).

   It is noteworthy that Cd concentrations in all tissues of Mo17 X H98
greatly exceeded those in corresponding tissues of Oh545 X B73 in every year
and for all sludge treatments including the controls. A 4x3x3x2 factorial
analysis of variance using 4 sludge rates, 3 corn tissues, 3 years and 2
hybrids showed that hybrid was the most significant main effect (P <_ .001) in
accounting for the variation in Cd concentration in corn tissue.  It is also
important to recognize that the grain-Cd concentration in the low (Oh545 X
B73) accumulator grown on the maximum-sludge-treated soils was only slightly
greater than that in the high (Mol7 X H98) accumulator when it was grown on
control plots with very low soil-Cd concentrations.

   Concentrations of Zn in leaves, grain, and stover of both hybrids are
presented in Table 37.  Higher sludge applications caused significantly
higher Zn concentrations within each year of all tissues of both hybrids.
Only stover produced by Mol7 X H98 showed a significant (P <_ 0.05) increase
in Zn concentration from year to year in response to accumulative sludge-
borne Zn applied during the study, although a trend for higher Zn contents in
Oh545 X B73 stover with additional annual applications of sludge-borne Zn
can also be seen.

   The two hybrids were selected to be different only in their capacities
to take up Cd and,  for the most part, this was accomplished.  However, during
the first and second years Mol7 X H98 on sludge-treated plots accumulated
more Zn in leaves and less in stover (P _<_ 0.01) than did Oh545 X B73.  Grain-
Zn concentrations were never significantly different for the two hybrids.
During all years, Mol7 X H98 had higher contents of Fe and Cu in grain, lower
contents of Mn in grain, higher contents of K and lower contents of Ca and Mg
in leaves than similar tissues from Oh545 X B73 (data not shown).

Conclusions

   Except for Fe, we are unaware of results showing changes in uptake of
metals by one genotype in response to stimuli produced by the roots of an-
other.  Wallace et al. (1962) grew two soybean genotypes (Glycine max L.)
that differed in their ability to accumulate Fe together in a solution cul-
ture containing a small quantity of calcareous soil.  They found that the
soybean variety that was inefficient in accumulating Fe caused a reduction in

                                      163

-------
 Taole  36. Concentrations (cry weight) of Cd  in  leaves, grain and  sr.over  of  two
           com hybrids grown on solic-plocs  of"  Elount silt  loam,  with  and
           without sewage sludge.
            Mo 17 X K98
                                                     OH545 X B73
                         Sludge  Irrigation Rates  (am)
Year
              6.4   12.?  25.4   LSD
0   6.u    12.7  25.4
LSD

1978
1979
1980

1973
1979
1 30

19'3
1979
19SO


0.852
0.981
0.927

0.056
0.084
0.090

0.753
1.22
1.45


8.27
8.80
9.52

0.626
0.563
0.974

5.78
10.3
14.2


12.6
18.7
21.3

1.10
0.943
1.12

10.8
23.9
24.9


20.2
37.2
42.0

1.70
1.43
1.33

24.6
34.7
44.4
^^
6
5
6

0
0
0

4
15
12
- ag/kg (dr
Leaves
.0**
.64*
. 42*
Grain
.247**
.19**
.316**
Stover
.52**
.5**
.4**


0.180
0.198
0.059

<0.062
<0.052
<0.062

0.163
0.271
0.253


1.71
1.51
0.845

<0.062
<0.062
<0.062

1.66
1.82
1.87


3.29
4.44
2.56

<0.062
<0.062
<0.052

2.33
4.18
3.53


6.39
12.0
6.98

0.131
0.109
0.095

8.48
11.5
13.2
-.
0.860**
4.74**
0.862**

0.041**
0.046*

1.56**
3.74**
b.22**
*» Significantly different at P<0.01
 •- Significantly different at P<0.05
£/ Not significantly different

-------
Table  37. CoriceTiTacic^s (dry weig'.-.t) of .In. ia leaves,  grain,  and
          slc-.'Si  o' :"o ccm hybrids sroun or. spli:-ploca  of Blour.c
          siii  losa, ;;ith ar.d WLCROUI sewage sludge.
ho 17 X K?S
Year
1573
1979
193C
1978
!979
.'Q30

!'.'•£
1979
15 KG
0
15.9
19.3
17.5
ii. 6
12.3
_ ? . 2

8.- '.5
V . -:'
12.2
&.4

56.5
51.6
54.2
29.9
16.9
26.7

34. 5
12.7

7~. 6
83.5
£9.3
3'.. -5
20.9
25. C

55 . ?
102
Ch545
Sludge Irrigation Races
25.4

148
173
12S
a:. 3
26.3
33.7

160
177
LSI

- - -ag
55.1-**
14.0*-*
25.7*-
5.70«*
3.90**
7.84**

16.5*-
13.J*-*
33.8*---

/kg fdry
Leaves
14
14
13
Orai".
19
12
15
SEover
5
10
G
wt ,
.5
.8
.7
.5
.8
.4

.12
.47
.4
X B73
(am)
6.4
\ _
39.9
44.7
49.2
26.3
21.0
23.5

32.9
36.5
51.7
12.7

61.3
73.0
76.0
31.2
31.2
29.4

79.3
93.2
109
25.4

118
139
130
3S.O
27.1
37.4

192
190
204
LSD

34. C**
35. S**
24.2**
8.40**
9.24**
9. 96**

54.9**
27.4*-*
25.5*«
**Sisrilf lean' lv dir.rerenc at PCO.Ol
                                   165

-------
Fe uptake by the variety that was efficient.  These results were confirmed
by Elmstrom and Howard (1969) from plants grown in a growth chamber in nu-
trient solutions containing various levels of Fe.

   If corn inbreds, having different inherited capacities to accumulate Cd
from contaminated soils, can interact to change uptake patterns of each other
when growing in close association, it was not obvious from the results of the
greenhouse study (Table 31).  Only where H98 and R805, a high- and low-Cd
accumulator, respectively, were paired in pots of sludge-amended soil was a
significant effect observed.  Cadmium concentrations in whole plants (34
days after planting) of R805 were higher when it was paired with H98 as com-
pared to concentrations when paired with itself or with B73, another low-Cd
accumulator.  However, Cd concentrations in leaves of R805 at the tasseling
stage of growth were not different regardless of which inbred it was paired
with.  This finding in R805 during early growth may represent a real root
interaction and deserves further investigation.  However, since this effect
was not general and occurred only during early stages of growth, we conclude
that the results of a study designed to determine gene effects affecting
genetic variations of Cd accumulations in corn would be biased very little,
if at all, by the proximity of plants growing in field plots.

   Reciprocal crosses between high-Cd and low-Cd and between high-Zn and
low-Zn accumulator inbreds showed no evidence of a maternal effect on the
uptake of metals.  Leaves from single-crosses contained concentrations of Cd
that were intermediate to those in leaves of their high- and low-Cd accu-
mulator parents, regardless of whether or not the parents were related.
Concentrations of Zn in leaves of single-crosses were also intermediate to
those contained in leaves of unrelated parent inbreds that accumulated
significantly different levels of the metals.  But where leaves of related
parent inbreds did not contain significantly different concentrations of Zn,
concentrations of the metal were lower in leaves from their single-crosses.
Lower Zn concentrations may have been due to a dilutive effect attributable
to hybrid vigor.  Dilution due to better growth of hybrids may complicate
determination of the relative importance of different gene effects that
control the accumulation of metals in corn plant tissues.

   Results from the three year field study showed that the two commercially
available hybrids selected for their different capacities to accumulate Cd
were indeed different when grown on split plots where soils contained various
levels of sludge-borne Cd.  At the time of selection it was thought that the
two hybrids would produce about the same grain yields.  Even though the low-
Cd accumulator produced more grain during the last two years, and more stover
in all three years than the high-Cd accumulator the magnitude of the dif-
ferences was small.  Differences in yields were also observed on control
plots and thus appeared to have no relationship to differences in capacities
to accumulate Cd.  At any rate the differences in yields were not enough to
account for such large differences in Cd accumulations in the two hybrids
by inferring a metal concentrating effect.  Furthermore, from previous analy-
sis of inbred parents of the two hybrids grown on sludge-treated plots, it
was estimated that the two hybrids would accumulate similar quantities of
other inorganic nutrient and non-nutrient elements,  especially Zn.  Although
there were small differences in amounts of some elements accumulated in par-


                                      166

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cicular tissues of the two hybrids, only Cd in leaves of Mol7 X H98 ex-
ceeded the maximum tolerable concentrations discussed by Allaway (1968)
and Melsted (1973).   However,  there were no overt symptoms of Cd toxicity in
either hybrid and no reason to expect that differences in yields were due to
anything more than inherent differences in growth potential.

   Differences in accumulations 'of Cd by the two hybrids did not affect Zn
uptake.  In two of the three years one of the hybrids accumulated more Zn in
leaves and less in stover than the other, but the magnitude of these differ-
ences was small relative to amounts accumulated in these tissues by both
hybrids growing on sludge-treated soils.  In the presence of an abundantly
available supply of  both Cd and Zn, the uptake of these metals appears to be
independent of each  other.

   For those concerned about enhanced levels of Cd in food chains,  it is
noteworthy that grain-Cd concentrations of Oh545 X D73 growing on maximum-
sludge-treated plots were significantly higher than those of Mol7 X H98
growing on control plots only during the first year.   Furthermore,  Cd concen-
trations in grain from Oh545 X B73 never exceeded the upper range of 0.294
mg/kg found in corn  grain of unidentified hybrids grown on conventionally
fertilized soils in  rural areas of Illinois (Pietz et al., 1978).  It is likely
that other combinations of parent inbreds could have been selected to produce
a hybrid not commercially available that would have minimized Cd accumu-
lations in grain produced on sludge-amended soils to levels not exceeding
the mean concentrations of 0.037 mg/kg found in corn grain produced on un-
adulterated soils.  One such hybrid might be B73 X R805.


EFFECT OF SOIL CATION EXCHANGE CAPACITY
ON THE UPTAKE OF CADMIUM BY CORN (Zea mays L.)

Introduction

   For sewage sludges containing more than 2 mg Cd kg of sludge, the U.S. EPA
(1979) regulated maximum accumulative applications of sludge-borne Cd on
soils with background pH values of 6.5 or higher according to cation exchange
capacities (CEC). Where soil cation exchange capacities are less than 5, in
the range of 5 to 15, and higher than 15 meq/100 g, maximum permitted accu-
mulative loading rates were, respectively, fixed at 5, 10, and 20 kg/ha of
sludge-borne Cd.  They stated that "several studies have demonstrated the
inverse relationship between CEC and plant uptake of Cd". However,  it appears
that most researchers have attempted to demonstrate the relationship by adding
soluble Cd salts to  soils.

   John et al. (1972) added 50 mg Cd as CdCl~ to pots containing 500 g of
each of 30 different soil types.  Using radish (Raphanus sativus L.) and
lettuce (Lactuca sativa L.) as test plants and linear regression analysis of
results, he concluded that the most important single factor associated with
lower levels of Cd in radish plant parts was the capacity of soils to adsorb
the metal.  Oxidizable organic matter contents of soils contributed to their
capacities to adsorb and therefore affected Cd concentrations in plants in-
versely.  Where the original organic matter of a clay soil was removed by

                                      167

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oxidation (H907 treatment) and replaced with muck to obtain different levels
of CEC,  Haghiri (1974) added constant amounts of Cd, as CdCl.,  in one study
and variable amounts in another to maintain a constant ratio of applied Cd
to CEC.   He grew oats (Avena sativa L.) for 4 weeks in pots to  determine Cd
uptake from the various soil mixtures.  He concluded that the decreased con-
centrations of Cd found with higher levels of added organic matter were pre-
dominantly due to its CEC.  That is, the Cd retaining power of  organic mat-
ter was due to its CEC rather than its capacity to chelate the  metal.  How-
ever, as Latterell et al. (1976) pointed out, when dry weight yields and
concentrations of Cd in oats, as reported by Haghiri (1974), were multiplied,
they showed a rather constant uptake of Cd regardless of soil CEC.  Miller
et al. (1976) grew soybeans (Glycine max L.) for 4 weeks in a CdCl2 rate ex-
periment in pots of 9 different soils selected to obtain a relatively wide
range of CEC, pH, and available P levels.  Where soils contained 1, 10, and
100 mg Cd/kg of soil, they found that Cd contents of plants were negatively
correlated (significant only at the 5% probability level) with  CEC only for
soils containing 10 mg Cd/kg.  Latterell et al.  (1976) diluted  Waukegan silt
loam soil with sand to obtain CEC's that ranged from 5.2 to 18.5 meq/100 g
and amended the mixture with digested sludge containing 2 mg Cd/kg of sludge
at rates equivalent to 0, 23.3, and 46.7 mt/ha (dry weight). Dry matter
yields and Cd concentrations in 30-day old soybean seedlings were unaffected
by differences in CEC.  Mean Cd concentrations in seedlings were slightly
increased by the 23.3 mt/ha sludge application,  but doubling the loading
rate did not cause further increases.  Sims and Boswell (1978)  mixed a "sec-
ondary industrial sewage sludge" with a Cecil loam soil (Typic  Hapludult) to
produce mixtures containing 0, 2.5, 5, and 10% sludge.  They superimposed
additions of 0. 5, and 10% calcium saturated bentonite over the sludge treat-
ments in a 3x4 complete factorial pot study.  Mixtures containing 5 and 10%
bentonite had initial CEC of 13.2 and 20.4 meq/100 g, respectively, as com-
pared to 7.4 meq/100 g in the untreated control soil.  Also, successively
higher loading rates of bentonite increased soil pH to 6.0 and  6.6 as compared
to 5.2 in untreated soil.  Sludge additions did not significantly alter the
CEC and soil pH.  They concluded that CEC and pH effects from added bentonite
could not be separated.  Much of the discussion of results concerned explana-
tion of an observed CEC reduction that occurred in bentonite mixtures between
the initial and post-harvest analyses.  The decrease in CEC with time was
directly proportional to amounts of bentonite added.

   In the above studies, soil cation exchange capacity was generally deter-
mined by the ammonium acetate method (Chapman, 1965), suggesting that all
exchangeable Cd was held by the same mechanisms.  Because of the conflicting
results reported by these investigators, it appears that the capacity of soils
to adsorb Cd and restrict its uptake by plants is complex and not controlled
directly by CEC.  It was the purpose of this study to determine whether plant
uptake of Cd is less when the source is sludge-derived than when the source
is the soluble CdCl_ salt and how plant uptake might be altered by differences
in soil CEC.

Methods and Materials

   The B., horizon (28 to 41 cm) of an Ava soil (Typic Fragiudalf), the Ap
horizon of a Maumee soil (Typic Haplaquoll), and the mixed A., and B,  horizons

                                      168

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(0 to 30 cm)  of a Plainfield soil (Typic Udipsamment) were collected to pro-
vide a wide range of organic-C contents and CEC's.  Results of the subsequent
textural analysis, performed on the bulk sample collected from an area mapped
as the Maumee series in Kankakee County, Illinois, indicated that this soil
was outside of the range for the Maumee series and was actually a mapping
inclusion.  Important characteristics of soil materials used in this study
are presented in Table 38.  Maumee Ap had a high CEC that was mainly due to
its organic mattei content   Ava B., had a moderately high CEC that was due
mainly to its content of clay and Hydrous oxides of Fe and Al.  Plainfield
has a low CEC, because it contained low concentrations of both organic mat-
ter and clay.  Ava was used in the study with and without additions of Plain-
field to obtain CEC of about 15.9, 10.6 and 5.3 meq/lOOg.  Plainfield was
also added to Maumee and to half and half mixtures of Ava and Maumee to ob-
tain a similar range of CEC's.  Mixtures of screened (7 mesh) and air-dried
soil materials were made in a twin shell soil blender.  To 12 kg of Ava and
each of 8 different soil mixtures, either air-dried and ground (10 mesh) di-
gested sewage sludge or a solution of 0.003 M CdCl. (applied by atomization)
was added at  rates (0.535 kg and 0.2477 g,  respectively) required to attain
a total Cd concentration of 10 mg/kg (dry weight) in soil or soil mixtures.
The sludge had an organic-C content of 12.8% (dry weight), CEC of 24 meq/100
g, pH of 6.0, and Cd concentration of 238 mg/kg.  Sludge was added at the
rate of 0.535 kg to supply a total Cd concentration of 10 mg/kg in 12 kg of
soil or soil  mixture.  Each of the 12-kg quantities of Ava or soil mixtures
were then subdivided into three-kg portions and each portion placed into one
of 4 plastic  pots (20.3 cm diameter).  Water was added to the pots until the
soil was completely saturated.  After drainage and evaporation had decreased
soil water contents to approximately field capacity, small cores of soil were
extracted with a probe (13 mm diameter) for analysis and eight evenly dis-
tributed corn seeds of the single-cross Mol7 x H98 were planted in each pot.
The pots were rotated once each week on the greenhouse bench.  The experi-
ment was conducted under natural sunlight supplemented with mercury-vapor
lighting to achieve a constant photo-period of L:D - 16:8.  Water was added
at the beginning and nutrient solution (commercial, 23-19-17) at the end of
each week to  replenish soil moisture contents.  Ten days after planting, the
number of plants was reduced to six per pot.  The first harvest of three


     Table  38. Characteristics of experimental soils.
     Soil
Particle Size Distribution
Sand       Silt        Clay    Organic-C
                                                              CEC
PH

Ava BI
Maumee Ap
P.'ainfield
	
11.9
30.9
93.9
v

60.4
53.5
3.4


27.7
15.6
2.7
	 2 	
0.24
16.3
0.25
meq/lOC g
15.9
58.9
l.i
units
4.6
7.2
5.3
                                      169

-------
above-ground whole plants was made three weeks after planting.   During the
first harvest the top-most leaves of the remaining three plants in each pot
were marked, so that when they were harvested at the end of seven weeks from
planting, new and old growth were separated for Cd analysis.

   After final harvest,  soil core samples were 'again taken from each pot
and analyzed separately for organic-C contents, CEC, Cd and pU.  In contrast,
samples collected initially from treatment replications were combined for
determining these 4 soil parameters.

   Soil and sludge parameters were determined as follows:   mechanical analy-
sis by the pipette method (Day, 1965);  organic-C by the Walkley-Black method
(Allison, 1965); CEC by the ammonium saturation method (Chapman,  1965) and
pH in a 1:1 soil or sludge to distilled water slurry employing a glass elec-
trode pH meter.  After drying (60 C), crushing, and pulverizing (60 mesh
screen), soil samples (0.5 g) were heated to 500 C for 24  hours,  digested in
concentrated HCL-HF and dissolved in 1 1J HC1 for determining total Cd con-
centrations, using atomic absorption spectrophotometry with background
correction.

   Plant samples were washed in distilled water immediately after cutting,
dried at 60 C, and ground in a Wiley mill to pass a 20-mesh screen.  Sub-
samples (2g) were wet-ashed in concentrated HNO, at 90°C,  taken to dryness,
and dissolved in 1 N^ HNO, for determining Cd concentration by atomic ab-
sorption spectrophotometry.

Results and Discussion

   Organic carbon contents, CEC's, Cd concentrations, and  pH levels of
sludge- and CdCl»-treated Ava 3..  and soil mixtures prior to planting and
after harvest of plants are shown in Table 39.  Except for Ava B., organic-C
contents were changed very little by sludge additions.  Maumee Ap contained
such a high organic-C content (16.3%) that the relatively  small amounts (0.6%
dry weight of soils) added as air-dried sludge could not be detected in mix-
tures that contained this particular soil.  Due to the inherent heterogeneity
of soils, sampling errors were evidently too high to detect the small de-
compositional losses of sludge-borne organic-C that may have occurred during
the 7-week study period.  In consideration of soil heterogeneity, the CEC's
were close to those anticipated for soil mixtures.  Although sludge no doubt
contributed to the CEC of sludge-treated soils, it was not a measurable ef-
fect.  No appreciable change in CEC's was observed for either sludge or CdCl_
treatments during the course of the experiment.  Total Cd  concentrations in
soils were near the proposed level of 10 mg/kg, except for sludge-treated
Ava.  Since the organic-C contents and CEC for sludge-treated Ava were within
expected ranges of values, this unforseen result was probably due to the non-
uniform distribution of Cd in sludge.  Differences of Cd concentrations in
soil samples collected prior to planting and those collected after harvest
were due to variability associated with sampling and analysis.   The highest
amounts of Cd extracted by plants were from the low CEC, Ava-Plainfield mix-
ture treated with CdCl2-  About 2.1% of the total Cd contained in this mix-
ture was removed as a constituent of harvested plants.  The amount removed
by plants was too low to detect in the soils due to the soil variability.


                                      170

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Table 39.  Initial and post-harvest (P-ll) characteristics of soil mixtsures amended with either sewage
          sludge or CdCl2-
Soil Mixture3

Ava
Ava-Plainf ield
Ava-Plainf ield
Ava
Ava-Plainf ield
Ava-Plainf ield
Maumee-A va-P la .
Maumee-Ava-Pla .
Maumee-Ava-Pla.
Maumee-Ava-Pla.
Maumee-Ava-Pla .
Maumee-Ava-Pla .
Maumee-Pla.
Maumee-Pla .
M.iumee-Pla.
Maumee-Pla.
Maiimee-Pla .
Maumee-P la.
Organic-C
Treatment

s ludge
sludge
sludge
CdCl2
CdCl2
CdCl2
sludge
sludge
s ludge
CdCl2
CdCl2
CdCl2
sludge
s 1 ud ge
sludge
CdCl2
CdCl2
CdCl2
Initial


0.70
0.69
0.43
0.24
0.17
0.17
3.34
1.76
0.89
3.86
1.79
0.64
3.83
2.53
1.19
3.94
2.79
1.06
P-ll,

Z____ 	
1.10+0.
0.84+0.
0.78+0.
0.37+0.
0.35+0.
0.33+0.
3.06+0.
2.03+0.
0.87+0.
2.51+0.
1.72+0.
0.64+0.
3.79+0.
2.08+0.
1.12+0.
3.32+0.
2.37+0.
0.87+0.

SDb


15
19
09
08
22
07
31
26
17
25
09
04
37
20
17
11
13
22
CEC
Initial
_ _ mni /
-— ——men/ '
17.7
9.4
3.6
16.1
8.6
3.9
18.2
10.2
5.4
18.1
10.9
5.8
16.2
10.4
6.2
17.7
11.3
3.7
P-ll, SD°
OOg 	
16.4+0.6
11. NO. 7
5.4+2.1
15.4+0.5
9.9+0.6
4.5+0.3
18.4+0.9
11.6+0.2
5.6^0.6
17.2+1.0
11.4+0.1
5.3+0.4
15.1+1.0
10.2+1.3
5.6+0.6
16.6+1.5
9.6+0.4
5.2+0.6
Soil-Cd
Init ial
	 mg/
19.6
13.4
8.0
10.5
9.2
11.6
12.2
10.7
9.0
11.9
11.4
10.0
9.3
9.5
7.0
12.8
13.3
9.8
P-ll, SD°
L «.
Kg 	
20.6+1.0
18.2+1.3
12.6+1.3
10.2+1.5
10.3+1.9
7.8+1.3
15.4+1.0
11.7+1.5
9.8+1.5
10.2+0.5
10.6+1.4
7.5+1.5
11.7+0.4
11.2+0.3
10.2^2.0
8.9+1.3
7.6+1.4
6.8+1.1
Initial

5.0
5.2
5.5
3.9
4.2
4.4
6.1
5.9
5.8
5.7
/ 5.8
5.8
6.6
6.5
6.3
6.6
6.6
6.4
pll
P-ll

4.8
5.2
5.5
3.7
3.7
4.1
5.6
5.7
5.7
4.9
5.1.
5.0-
6.0
6.1
6.0
6.2
6.1
5.6

SDb

0.1
0.1
0.1
0.1
0.03
0.03
0.1
0.1
0.1
0.2
0.2
0.04
0.
0.
0.
0.
0.
0.
 "Each  set  of three soils represents three different dilutions with Plainfield Sand estimated to yield
   CEC  levels of approximately 5.3,  10.6 and 15.9 meq/100 g.
'SD
Standard Deviation

-------
The Ava soil pH (Tables 38 and 39) was increased by sludge and decreased
by CdCl. creacmencs.  Sludge was more and CdCl. less effective in changing
the pH with higher dilutions of Ava B. with Plainfield.  Ava and Ava-Plain-
field mixtures had lower pH's than otner mixtures.  At the two highest CEC's,
Maumee-Plainfield mixture had higher pH values than other mixtures.  How-
ever, except for Ava and Ava-Plainfield mixtures, pHlevels within the range
of low to high CEC's were not different nor were they different between the
sludge and CdCl. treatments.

   Data presented in Figure 5 clearly depicts the presence of an inverse
relationship between Cd uptake by corn plants and soil CEC where the source
of Cd was a soluble salt,  but not where it was added as a constituent of
sewage sludge.  The high F ratio, as shown in the analysis of variance for
whole plant data (Table 40), leaves little doubt that the main difference
in Cd uptake was due to source.  The second most important factor which af-
fected Cd uptake was differences in soil mixtures.  Soil CEC exerted the
least influence on Cd uptake.  All interaction effects were highly signifi-
cant (P _< 0.01).  The most important interaction was between Cd source and
soil mixture.  When single degree of freedom comparisons were made (Table
40), it was found that both the linear and quadratic components for regression
of Cd uptake on CEC were highly significant (P <_ 0.01) for CdCl. source, ex-
cept the quadratic component for the Maumee-Ava mixtures.  This analysis con-
firms that sludge controls plant uptake of Cd more than does CEC.  Uptake of
Cd from Ava and Ava mixtures with Plainfield was significantly (P £ 0.01)
higher than that from Maumee mixtures with Plainfield at all levels of CEC
when the source of Cd was CdCl..  But, when the Cd was sludge-derived, uptake
was significantly higher (P _< 0.05) from Ava than Maumee mixtures only at
the highest level of CEC.   However, this difference between sludge-treated
Ava and Maumee mixture was undoubtedly due to the anomalously high Cd con-
centration in Ava (Table 39) as discussed previously.  Cadmium concentrations
in whole plants from the Ava-Maumee mixtures, amended with CdCl. was not sig-
nificantly different than means amounts calculated from the individual Ava
and Maumee series, except for mixtures having low CEC.  This result can be
confirmed visually from an examination of Figure 5.

   Since soil pH and organic matter were both higher according to the order
Maumee > Maumee-Ava > Ava mixtures, their effects on Cd uptake from CdCl-
additions cannot be separated and evaluated from these results.  However, in
studies discussed previously in this report where repeated applications of
digested sludge were made on plots of an acid silt loam soil and calcareous
strip mine spoil material for periods of 12 and 7 years, respectively, Cd
uptake by corn was considerably higher on spoil and continued to increase
with repeated sludge applications.  This was true even though pH was much
higher in the calcareous spoil than that of the acid silt loam.  If pH ex-
hibited a dominant influence on Cd uptake, the sludge-amended Ava series
should have produced plants with higher Cd levels than all other soil mix-
tures, except the Ava series amended with CdCl..  However, Cd uptake from
the sludge-amended Ava series was not significantly different, with one ex-
ception, from the Maumee series that had the highest pH, indicating that
sludge properties may be more important.  The exception, as previously ex-
plained, was caused by a higher soil-Cd concentration.
                                      172

-------
LO
       350
       300
       250
     O)

     - 200
       150
       100
        50
         0
    T-    T-
. FIRST HARVEST,
   Whole Plant
                                                b. SECOND HARVEST,
                                                   Plant Bottom
                LOW
                        MED.
                                                      I
 c. SECOND HARVEST,
      Plant Top
EXPLANATION
Cd Source
SOIL MIXTURE
Ava
Maumee/Ava
Maumce
CdCI2
A
0
a
Sludge
A
•
•
                                                                      20°
15 ^^ 20 0
  HIGH         'LOW'   'MED.'      HIGH
      CATION EXCHANGE CAPACITY (meq/100 g)
LOW
                                                                   MED.
                                                                                                     20
                                                                                                HIGH
           Figure 5.  Concentrations of Cd in corn tissue grown on soil mix combinations resulting in
                     3 levels of CEC, containing 10 mg/kg of Cd derived from CdCl2 or sewage sludge.

-------
Table  40. Analysis of variance for Cd concentrations in plants (mg/kg)
          harvested three weeks after planting.
Source of Variation    df          SS           MS            7
A - CSC
B • soil mixture
C • Cd source
AS
AC
BC
ABC
Error
Total
2
2
1
4
2
2
4
54
71
24,247.93
109,573.52
374,186.03
9,087.31
16,776.34
75,332.71
10,166.09
6,469.68
625,639.61
12,123.97
54,786.76
374,186.03
2,271.83
8,388.17
37,666.35
2,541.52
119.81

101.19**
457.28**
3,123.19**
18.96**
70.01**
314.39**
21.21**


Significance  levels  for  single  degree  of  freedom comparisons

	Cpaoarison                            Cd Source
                                                        Sludge     CdCl2

CEC.  (linear) within Ava                                n.s.        **
CECQ  (quadratic) within Ava                             n.s.        **
CEC£ within M/A  (Maumee/Ava)                            n.s.        **
CEC  within M/A                                         n.s.       n.s.
CEC" within Mauaee                                      n.s.        **
CECjT within Maunee                                      n.s.        **

[Ava vs. Maumee] within CEC-                            n.s.        **
[M/A vs. mean (Ava + Maumee)] within CECL               n.s.        **

[Ava vs. Maumee] within CEC.                            n.s.        **

[M/A vs. mean (Ava + Maumee)] within CEC               n.s.      .n.s.
[Ava vs. Maumee] within CEC.,                              *          **
[M/A vs. mean (Ava + Maumee)] within CEC               n.s.       n.s.
                                         a


*, ** «= significant at P^O.05 and  P<0.01,  respectively.
n.s. = non-significant F-ratio.
                                    174

-------
   An analysis of variance was performed on Che data presented in Figures
5b and 5c for Cd concentrations in the bottom and top portions of plants
harvested seven weeks after planting.   Results of the data analysis were
similar to those for the first cutting (Table 40) except that concentrations
of Cd in plants from Maumee-Ava mixtures were significantly different from
the mean concentrations calculated from individual series of Ava and Maumee,
at all CEC levels.  The data plotted in Figures 5a and 5b showed that Cd con-
centrations in the bottom portion of plants from the CdCl_ treated Ava series
remained about the same as that of whole plants from the first harvest.
Because the first growth on the CdCl.  treated Ava series showed symptoms of
severe Cd toxicity, further growth or  the bottom portion of these plants was
almost nil.  Cadmium toxicity affected further growth of the bottom portion
of plants between the first and second harvest on CdCl- treated Ava-Maumee
and Maumee mixtures to a lesser extent than those on the Ava series so that
Cd concentrations may have decreased as a result of the diluting effect  of
additional growth on these two.  Concentrations of Cd in the new growth
material produced during the interval  between the first and second harvest,
were less on all soil mixtures than in plants at the first harvest (Figure
5, a and c).  This may have been due to decreasing availability of Cd in soil
mixtures and/or decreasing uptake of Cd with plant age.

   Toxicity symptoms of chlorotic streaking and stunting were visually evi-
dent for the CdCl. treatments; symptoms were less severe according to the
order Ava, Ava-Maumee, and Maumee series.  Where Cd was supplied as a con-
stituent of sludge, no overt symptoms  of Cd toxicity were observed in plants
grown on any of the different soil mixtures.  The single-cross (Mol7 x H98)
used in this study was selected for its inherited capacity to accumulate high
concentrations of Cd without showing toxicity symptoms on a soil that con-
tained about 3-fold higher concentrations of sludge-borne Cd than used in
this study (Hinesly et al., 1981a). At the approximately 10 mg/kg of soil-Cd
used in this study, differential Cd uptake was probably due to factors ex-
ternal to plant roots affecting availability of the metal, rather than plant
physiological processes regulating the absorption and translocation of Cd.

   Mean weights (dry) of plants grown  for 3-weeks on the several soil mix-
tures treated with CdCl. and sludge are shown in Figure 6a along with mean
amounts of Cd accumulated per plant (Figure 6b).  An analysis of variance
showed differences in weights were due mainly to Cd source and to a lesser
extent by soil mixture.  But, soil CEC and interaction effects were not  sig-
nificant.  Plants appeared to make normal growth on all sludge-treated soil
mixtures, while those on all CdCl. treated mixtures were stunted and showed
typical Cd toxicity symptoms.  The CdCl- treatment affected plant growth to
a lesser extent (Figure 6a) on Maumee  than on Ava mixtures.

   The analysis of variance for mean total amounts of Cd per 3-week old
plants is presented in Table 41.  By order of listing, differential uptake
of Cd was affected by Cd source, soil  mixture, and soil CEC.  Since all
interactions were significant, single  degree of freedom comparisons were
made.  These showed that total amounts of Cd accumulated per plant were  not
different where the metal source was sludge, but did differ in response  to
these parameters where CdCl_ was the source (Table 41 and Figure 6b). On
the CdCl~ treated Ava and Maumee series total amounts of Cd accumulated  de-


                                      175

-------
creased in a linear fashion as CEC increased.  At all levels of CEC,  plants
accumulated about 2-fold higher total amounts of Cd when grown on the Ava
series treated with CdCl« as compared to amounts from Maumee treated with
equivalent amounts of the salt.  Except at low CEC, plants grown for 3 weeks
on CdClj-treated Maumee-Ava mixtures accumulated less total Cd than would
have been predicted from the average amounts accumulated from fhe individual
Ava and Maumee series.

   Where corn plants were grown for 7 weeks, average weights of plants were
significantly affected to the greatest extent in order of Cd source,  soil
mixture, and CEC (Table 42 and Figure 7a).  All interactions were significant.
Single degree of freedom comparison showed that plant weight increased with
increased CZC on the CdCl» treated Maumee-Ava and Maumee series,  but not on
the CdCl2-treated Ava.  Plant weights were unaffected by differences in CEC's
of soil mixtures where Cd was supplied as a constituent of sludge.  Plant
weights on different sludge-treated soil mixtures were not significantly dif-
ferent.  But, on the CdCl_-treated Ava and Maumee series, plant weights were
significantly higher on the latter at all CEC levels, except the lowest.

   Total amounts of Cd accumulated per plant at the end of 7 weeks of growth
on sludge and CdCl_ treated soil mixtures are shown in Figure 7b  and the
analysis of variance for these data is presented in Table 43.  All three
main parameters and their interactions significantly influenced the total
amounts of Cd accumulated per plant.  The significance of CEC and inter-
action effects that influenced total Cd accumulations in 7-week old plants
were contrary to results from 3-week old plants.  For older plants grown on
CdClj-treated soil mixtures total amounts of Cd accumulated per plant de-
creased as soil CEC increased, but total accumulations of sludge-borne Cd
were unaffected by CEC.  At all CEC levels plants grown on the Ava series
accumulated significantly more total Cd than those on the Maumee series for
both sources of Cd.  However, amounts accumulated were markedly less on all
sludge-treated soil mixtures than they were for CdCl. treatments, except at
the high CEC level for CdCl.-treated Maumee and sludge-treated Ava, where
amounts were about the same.  This was probably a reflection of the anom-
alously high concentration of sludge-borne Cd in Ava, as discussed earlier.
Total amounts of Cd accumulated by corn on CdCl.-treated Maumee-Ava mixtures
were less than mean amounts calculated from results of similarly treated Ava
and Maumee, except at the high CEC level.  At all levels of CEC,  total ac-
cumulated amounts of Cd from sludge-treated Maumee-Ava mixtures could have
been predicted from the mean of amounts accumulated per plant on the sludge-
treated Ava and Maumee series.

   When considered separately, concentrations of Cd in and weights of plants
grown on the three different series of sludge-amended soil mixtures were not
significantly different, but were different when these data were used to cal-
culate total amounts of Cd accumulated per plant.  Except to increase CEC,  it
appeared that soil organic matter did not play a dominant role in controlling
Cd accumulation by corn.  In the CdCl.-treated Ava, organic matter contents
were affected very little by diluting with Plainfield (Table 38)  even though
amounts of Cd accumulated by 7-week old corn plants were reduced by about 32%
(Figure 7b).  In the CdCl^-treated Maumee, organic matter contents were re-
duced by about 74% by diluting with Plainfield, but amounts of Cd accumulated


                                      176

-------
   1.8



   1.8



   1.4



- 1.2
X

at
5 1.0
H


^ 0.8
>  0.6
0.4
   o.:
             a.  FIRST HARVEST
             EXPLANATION
                         Cd Source
      SOIL MIXTURE  CdCl;  Sludge

      Ava            A     A
      Maumee/Ava     o     •
      Maumee         a     •
            |  5  |
             LOW
,  10  |
 MED.
                                         0.18
                                       0.16
                                          0.14
                                          0.12
                                          0.10
                                          0.08
                                        •3
                                        CJ
                                       0.06
                                          0.04
                                       o.o:
                                      20
                                                  b.  FIRST HARVEST
                                          "o    ".  S  .   .  10  .
                            HIGH                 LOW     MED.

                 CATION EXCHANGE CAPACITY  (meq/100 g)
                                                                     15 | _ (  20
                                                                        H1GH
      Figure 6.  Mean weight  (a) and  amount of Cd accumulated (b) per plant
                 for corn grown for 3 weeks on mixtures of soils combined  to
                 give 3 levels of  CEC,  containing 10 mg/kg of Cd derived
                 from CdCl. or sewage sludge.  - -
                                 177

-------
Table 41.  Analysis of variance for mean total amounts  (ing)  of  Cd  accumu-
          lated per plant during the first three weeks.
Source of Variation    df         SS           MS
A = CEC
3 • soil mixture
C « Cd source
AB
AC
3C
ABC
Error
Total
2
2
1
4
2
2
4
54_
71
0.0036
0.0126
0.0570
0.0014
0.0010
0.0048
0.0016
0.0066
0.0886
0.0018
0.0063
0.0570
0.0003
0.0005
0.0024
0.0004
0.0001

14.95**
51.83**
468.75**
2.81*
3.92*
19.84**
3.41*


Significance levels for single degree of freedom comparisons

                      Comparison                           Cd Source

CECT (linear) within Ava
CSC!* (quadratic) within Ava
CEC;* within M/A (Maumee/ Ava)
CECT within M/A
CEC£ within Maumee
CEC^ within Maumee
Q
[Ava vs. Maumee] within CEC-
[M/A vs. mean (Ava + Maumee)] within CEC.
[Ava vs. Maumee] within CEC
[M/A vs. mean (Ava + Maumee)] within CEC
[Ava vs. Maumee] within CEC
[M/A vs. mean (Ava + Maumee)] within CEC
Sludge
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.

n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
CdCl2
**
n.s.
n.s.
ir.s.
**
n.s.

**
n.s.
**
*
**
**
*, ** = significant at P<0.05 and P^O.Ol,  respectively.
n.s. = non-significant F-ratio.
                                      178

-------
Table 42.  Analysis of variance for total dry weight (g) in plants
           after seven weeks of growth.
Source of Variation    df         SS           MS
A = CSC
B =• soil mixture
C a Cd. source
AB
AC
BC
ABC
Error
Total
2
2
1
4
2
2
4
54
71
18.6336
19.6253
427.2939
7.5589
7.2003
44.3036
8.9672
28.1700
561.75
9.3168
9.8126
427.2939
1.8897
3.6001
22.1518
2.2418
0.5217

17.86**
18.81**
819.09**
3.62**
6.90**
42.46**
4.30**


Significance levels for single degree of freedom comparisons

                      Connarison                           Cd Source

CEC (linear) within Ava
CEC (quadratic) within Ava
CEC:* within M/A (Maumee/Ava)
CECT within M/A
CEC£ within Maumee
CEC within Maumee
[Ava vs. Maumee] within CEC,
[M/A vs. mean (Ava + Maumee)] within CEC.
[Ava vs. Maumee] within CEC
[M/A vs. mean (Ava •)- Maumee)] within CECU
M
[Ava vs. Maumee] within CEC_
[M/A vs. mean (Ava -I- Maumee)] within CEC
Sludge
n.s.
n.s.
n.s.
n.s.
n.s.
*
n.s.
*
n.s.
**
n.s.
**
CdCl2
n.s.
n.s.
*
n.'s.
**
n.s.
n.s.
*
**
**
**
n.s.
*, ** = significant at P<0.05 and P^O.Ol, respectively.
n.s. « non-significant F-ratio.
                                     179

-------
   18
   16
   14
•S 12
K-

3
LU
5 10
 LU
 O
 <
 (X
 01
    4
           a.  SECOND HARVEST
   EXPLANATION
              _Cd Source

SOIL MIXTURE
Ava
Maumee/Ava

Maumee
CdCi.

 A
 o

 a
                             Sludge

                              A
            LOW
                                          0.9
                                 0.8
                                          0.7
                                 0.6
                               c.

                               "a.
                                 0.5
                                          0.4
                               •3
                               0
                                          0.3
                                          0.2
                                           0.1
                                           b. SECOND HARVEST
                     10  ,     15 ,   ,  20       0     i	5,    . 10 -i     15 ,      20
                    15 |	. 20             	
           MET       HIGH                'LOW'    'MED.        HIGH
             CATION EXCHANGE CAPACITY (meq/100 g)
     Figure  7.  Mean weight  (a)  and  amount  of  Cd accumulated (b)  per plant
                for corn  grown for 7 weeks  on  mixtures  of  soils combined to
                give 3  levels of CEC,  containing- 10 mg/kg  of Cd derived
                from CdCl or sewage sludge.
                                180

-------
Table 43.  Analysis of variance for mean total  amounts  (mg) of Cd
          accumulated per plant during  seven weeks of  growth.
Source of Vaziacicn    df         SS           MS            F
A = CSC
B =» soil mixture
C » Cd source
AB
AC
BC
ABC
Error
Total
2
2
1
4
2
2
4
54
71
0.0771
0.9780
1.8556
0.0152
0.0937
0.3779
0.0213
0.0729
3.4917
0.0386
0.4890
1.8556
0.0038
0.0468
0.1890
0.0053
0.0014

28.57**
362.20**
1,374.42**
2.81*
34.70**
139.96**
3.94**


Significance  levels  for  single  degree  of  freedom comparisons

                       Comparison	Cd Source

CZC_ (linear) within Ava
CZC (quadratic) within Ava
CZC? within M/A (Maunee/Ava)
CZC" within M/A
CEC£ within Maumee
CEC within Maumee
[Ava vs. Maumee] within CZC_
[M/A vs. mean (Ava +• Maumee)] within CZC.
[Ava vs. Maumee] within CZC..
[M/A vs. mean (Ava + Mauaee)] within CZC
[Ava vs. Maumee] within CZC..
[M/A vs. mean (Ava + Maumee)] within CZC
n
Sludge
n.s.
n.s.
n.s.
n.s.
n.s.
n.s.
**
n.s.
**
n.s.
**
n.s.
CdCl2
**
**
**
'*
**
n.s.
**
**
**
**
**
n.s.
 *,  ** -  significant at P£0.05 and P<0.01,  respectively.
 n.s.  = non-significant F-ratio.
                                    181

-------
by plants decreased from low to high CEC's by only 36%.  Therefore,  within a
particular series, organic matter affected the amounts of Cd accumulated per
plant very little, except for its contribution to additional soil CEC.  Ap-
parently, the difference in amounts of Cd accumulated by corn plants from
different series of soil mixtures was controlled more strongly by some other
factor than explained by differences in soil organic matter contents.

   Cadmium contained in the digested sewage sludge must have been present in
forms that inhibited its uptake to such an extent that soil difference in
organic matter contents, CEC, pH, and particle size distribution were rela-
tively unimportant.  On the other hand, municipal sewage sludge may have
supplied other substances that suppressed the availability of Cd or antag-
onized its uptake.

Conclusions

   1.  Generally, soil CEC inversely affected the uptake of Cd by corn and
its growth when Cd was supplied as a soluble salt, but not when the Cd was
supplied as a constituent of municipal sewage sludge.  Thus, source of Cd
was the most important factor affecting its uptake and impact on plant growth.

   2.  Where soil mixtures were amended with CdCl_, tissue-Cd concentrations
were higher and plant growth lower on soil mixtures that had low organic
matter contents, at all equivalent levels of CEC.  However, within a particu-
lar series of mixtures there was no evidence that soil organic matter de-
creased Cd uptake beyond its effect on soil CEC.  Tissue-Cd concentrations
and plant growth on sludge-amended soil mixtures were not significantly dif-
ferent regardless of soil CEC or its origin.  Except for the series of soil
mixtures with the lowest organic matter content (Ava B..), sludge applied at
a rate of 100 me/ha did not significantly affect soil organic-C concentrations.
Thus, the lack of a relationship between soil CEC and Cd concentrations in
plants on these soil mixtures was not solely attributable to concomitant ad-
ditions of sludge organic matter.

   3.  Total amounts of Cd accumulated per plant, as calculated from tissue-
Cd concentrations and dry weight of harvested tissue, also varied inversely
with soil CEC on CdCl-2treated soil mixtures but not on those treated with
sludge.  However, total amounts of Cd accumulated per plant were signifi-
cantly higher on sludge-treated Ava mixtures where CEC was due less to or-
ganic matter contents than other sources.

   4.  The results of this study indicate that the information obtained from
studies using soluble salts of Cd cannot be extrapolated to predict potential
hazards from sewage sludge, and perhaps other waste applied on land as a means
of disposal.  Further research is needed to determine why sludge-borne Cd was
taken up to a lesser extent than that applied as a soluble salt.  To explain
why amounts of Cd accumulated by corn plants differed with respect to the
dominant origin of soil CEC will require information not presently available.
                                      182

-------
UPTAKE OF METALS BY SPINACH GROWN ON SOIL AMENDED WITH SLUDGE AND CdCl2

Introduction

   Since many investigators have undertaken metal uptake studies with sludges
artificially spiked with high levels of Cd salts', an experiment was designed
to determine if the addition of a chemical source of cadmium (CdCl_) to
sludge produces the same uptake of metal as that of indigenous sluage-Cd.
Spinach was chosen to evaluate Cd uptake because of its known high accumula-
tion of the metal in its leaves.

Materials and Methods

   A total of ten different sludges were used in the experiment; two each
containing approximately 40, 140, 400,  600, and 1000 mg Cd/kg (dry weight).
For the first 4 Cd levels,  one sludge of each pair contained completely in-
digenous Cd; the other sludge contained a lower level of indigenous Cd and
was spiked with CdCl_ to raise its Cd content to that of the other member
of the pair.  Both sludges  containing 1000 mg Cd/kg had additions of CdCl_,
but they differed in their  initial indigenous content.  Table 44 shows the
Cd contents of the ten sludges as measured by atomic absorption spectro-
photometry after their preparation.

   Each of the ten sludges  were admixed with appropriate amounts of Blount
silt loam (Aerie ochraqualf, fine illitic, mesic, pH 7.4) to produce 9 kg
of soil at each of the following loading rates: 10, 5, 2.5 and 1.25 kg Cd/ha.

   The 10 kg Cd/ha rate corresponds to a soil level of 4.46 mg/kg Cd.  An
amount of liquid sludge was added to dry Blount silt loam to produce 18 kg
of dry material at the 4.46 mg/kg rate for each of the ten sludge treatments.
Each mixture was air dried  on a sheet of Visqueen and stirred frequently.
Portions of each were then  mixed with untreated soil to produce the other
rates by the following scheme:  1.25 kg Cd/ha (.558 mg/kg) = 1.12 kg mix +
7.88 kg soil; 2.5 kg Cd/ha  (1.12 mg/kg) = 2.25 kg mix + 6.75 kg soil; 5.0
kg Cd/ha (2.23 mg/kg) = 4.5 kg mix + 4.5 kg soil; 10.0 kg Cd/ha (4.46 mg/kg)
= 9 kg mix.  Aliquots of soil at the four rates containing only CdCl_ (ap-
plied by atomization) and one aliquot for control were also prepared.

   After addition of N-P-K  at the equivalent rate of 100 kg/ha, each aliquot
of soil was thoroughly mixed in a twin shell soil blender and equally dis-
tributed to three eight-inch pots (3 kg soil/pot).  Pots were arranged on
three greenhouse benches in a completely randomized design.  Approximately
10 seeds of Bloomsdale spinach were planted in each pot on 25 April 1980.
After germination, the number of plants per pot was reduced to four.

   Harvesting of plants began 24 June.   Plants were cut when they had reached
approximately commercial-sale size.  Each sample was taken through three
washings of distilled water, dried at 60°C, and ground in a Wiley mill to
pass a 20-mesh screen.  Plant samples (2g) were wet ashed in concentrated
HN03 at 90 C followed by concentrated HCIO^ at 200°C, taken to dryness, and
dissolved in 1 I) HNO-.  Analyses for 13 chemical elements were done by atomic
absorption spectrophotometry with appropriate background correction.

                                      183

-------
        TABLE 44.   TOTAL SOLIDS  AND CADMIUM CONTENT OF THE  TEN SLUDGES  USED IN THE  GREENHOUSE
                  SPINACH  EXPERIMENT.
oo
Cd concentrations (rag/kg
Sludge
Number
1
2
3
4
5
6
7
8
9
10
Date
Collected
Dec. 1979
Dec. 1979
Dec. 1979
Dec. 1979
July 1979
July 1979
July 1979
July 1979
Dec. 1979
Dec. 1979
Source
HPLa
HPL
11 PL
HPL
LLb#23
LL #29
LL #20
LL #29
HPL
HPL
% of
Solids
10.2
7.50
7.56
7.57
14.4
8.7
17.7
8.82
7.13
9.58
original
41
18
166
18
356
218
541
218
166
41
after CdCl2
addition
41*
40
166*
159
356*
398
541*
594
1020
1120
dry weight)
proposed
40
40
140
140
400
400
600
600
1000
1000
        *  No CdCl2  added

        aHPL = Hanover  Park  Lagoon

        LL  = Lawndale Lagoon

-------
Results

   Data were analyzed by a two factor analysis of variance using three Cd
sources and four loading rates for each of the five different levels of
cadmium.  Elemental levels in spinach tissue and statistical analyses are
summarized in Tables 45-49.  It was deemed inappropriate to use Cd level
as a third factor,since within the sludge + CdCl? treatment, the varying
Cd levfds were achieved by different proportions'of sludge and CdCl.;
analyses were therefore performed separately.

Discussion

   The only elements that had consistently significant differences in up-
take by spinach for both loading rates and Cd source were Cd and Zn.  Sig-
nificance for Zn uptake is certainly not surprising since appreciable amounts
of it were added as components of sludge and these amounts, unlike Cd which
was controlled, varied with the amount of sludge.  Thus, Zn uptake generally
decreased in the order of sludge > sludge + CdCl,, > CdCl- because amounts
of total soil Zn decreased.

   Cd levels in spinach generally increased in the order of sludge < sludge +
CdCl- < CdCl-.  This suggests that Cd is less available when applied as a
constituent of sewage sludge than when applied as a soluble salt.  However,
the interaction of loading rate and Cd source was significant in all cases
indicating complexities in the relationship which are not presently understood.


RESPONSES OF WHITE LEGHORN CHICKENS TO BIOLOGICALLY INCORPORATED CADMIUM

Introduction

   Concentrations of several heavy and transition metals were increased in
the foliage and grain of crops grown on soil amended with digested sewage
sludge (Hinesly and Hansen, 1979).  The most notable enhancements of metal
concentrations in plant tissues were those of Zn and Cd.  Except for Cd,
most of the trace elements whose concentrations were increased in plant tis-
sues by sludge applications are known to be essential elements for animals,
and rations are frequently supplemented with mineral additives to provide
them at levels required by modern breeds of animals.  Cadmium has not yet
been shown to be essential for animal health.  However, it has been reported
at low concentrations to be beneficial for the growth of rats (Schwarz and
Spallholz, 1978).  Short time studies where pheasants and swine were fed
corn grain from soils with and without sewage sludge additions demonstrated
that higher concentrations of grain-Cd caused increased Cd concentration in
animal livers and kidneys without adversely affecting performance or health
(Hinesly et al., 1976; Hansen et al., 1976; Hansen and Hinesly, 1979; Hinesly
et al., 1979).  It has been shown that Cd biologically incorporated into corn
grain and soybeans is nearly as available for absorption by animals as that
added to rations in the form of CdCl. (Buck et al., 1979).  But, soluble Cd
salts have generally been added to animal rations at much higher concentra-
tions than were attainable from plant materials alone.  Furthermore, single
ion additions create a nutrient imbalance that generally can not be duplicated
by using plant materials.

                                      185

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        TABLE 45. CONCENTRATIONS OF SELECTED CHEMICAL ELEMENTS (DRV WEIGHT) IN SPINACH GROWN ON B1OUNT SILT LOAM AMENDED  WITH

                  SEWAGE SLUDGE. SLUDGE PLUS CdCl2 AND CdCI2 AT RATES TO PROVIDE EQUIVALENT AMOUNTS OF TOTAL SOIL-CD.   SLUDGE

                  AND SLUDGE PLUS CdCl2 CONTAINED 40 ing/kg (DRY WEIGHT) OF Cd.
00
o
Cd-Source
Sludge
Sludf.e *
CdCl2
CdClj
Cd-Loading Rate
1.25
2.50
5.0
10.0
1.25
2.5
5.0
10. 0
1.25
2.5
5.0
10.0
K

5.42
5.51
5.72
6.53
6.30
5.00
5.87
5.74
6.86
6.75
5.88
5.01
Na

0.715
0.671
0.8SO
0.913
0.561
0.692
0.782
0.800
0.574
0.544
0.718
0.636
Ca

2.38
2.48
2.61
2.95
2.10
2.49
2.91
3.50
1.89
1.66
1.93
2.04
Me

2.04
1.98
1.82
1.50
2.14
2.20
1.93
1.34
1.98
1.93
1.96
2.46
Fe Mn

828.00 93.17
528.00 115.83
591.67 255.87
254.00 874.00
381.33 110.57
839.33 127.97
444.33 367.20
279.67 861.13
739.33 115.17
564.67 103.63
843.33 122.27
799.00 147.77
Zn Cd

259.27 5.82
480.43 10.22
520.57 15.13
532.40 15.13
275.67 7.53
423.70 13.95
512.30 20.99
544.20 24.73
116.90 4.97
115.57 7.87
112.97 21.81
115.70 61.19
Cu

">g/ Kg
22.77
30.92
40.78
48.45
26.98
36.73
42.80
48.74
17.58
25.48
20.40
17.61
Hi

1.43
2.32
1.67
6.32
0.93
1.67
2.50
7.74
1.29
2.09
1.71
1.33
Cr

3.70
4.53
4.95
3.74
5.25
5.39
11.03
3.42
2.93
2.28
2.74
2.60
I'b Al

4.42 1203.33
1.68 730.00
2.04 837.33
2.37 450.33
1.51 465.67
1.97 1284.67
2.37 586.00
2.15 539.00
1.82 1312.33
1.55 962.00
1.47 1248.33
1.B9 1298.67
F Fatio Significance Levels
Loading
Rate
Cd Source
Interaction
n.s.
n.a.
*
**
**
n.a.
**
**
n.s.
n.a.
*
**
n.a. **
n.a. **
n.a. **
** **
** **
** **
**
**
**
**
**
**
*»
**
**
n.a. n.a.
n.a n.a.
n.a. n.a.
        *. **=Significantly different at P£0.05 and P£0.01,  respectively.



        n.s.-No significant differencea.

-------
       TABLE  46. CONCENTRATIONS OF SELECTED CHEMICAL ELEMENTS (DRV WEIGHT) IN SPINACH  CROWN ON  BLDUNT SILT  I0AM AMENDED WITH
                 SEWAGE SLUDGE. SLUDGE PLUS CdCl2.  AND CdCI2 AT RATES TO PROVIDE EQUIVALENT AMOUNTS OP TOTAL SOIL Cd.  SLUDGE
                 AND SLUDOfc PLUS CdCl2 CONTAINED UO mg/kg (DRY WEIGHT) OF Cd.
CO
Cd-Source Cd-Loading Rate
Sludge
Sludge +
CdCl2
CdCl2

1.25
2.5
5.0
10.0
1.25
2.5
5.0
10.0
1.25
2.5
5.0
10.0
P Ratio Significance Levels
Loading Rate
Cd Source
Interaction


K

6.42
7.04
7.10
6.47
6.78
5.73
5.47
5.95
6.86
6.75
5.88
5.01
n.s.
*
n.s.
Na
•• z
0.644
0.452
0.781
0.924
0.547
0.766
0.602
0.742
0.574
0.544
0.718
0.636
*
n.a.
*
Ca

2.03
2.00
2.21
2.45
1.77
2.59
3.00
2.60
1.89
1.66
1.93
2.04
**
**
*
MB

1.96
1.86
1.82
1.98
2.10
1.86
2.02
2.08
1.98
1.93
1.96
2.46
n.a.
n.a.
n.a.
Fe Hn

554.33 97.20
723.67 98.77
456.67 92.80
636.33 127.23
461.00 83.07
1575.33 106.80
995 33 120.43
470.67 62.97
739.33 115.17
564.67 103.63
843.33 122.27
799.00 147.77
n.a. n.s.
n.a. **
n.a. **
Zn Cd

144.77 9.05
171.70 11.92
199.17 14.00
336.80 28.54
166.97 12.26
155.17 13.73
277.57 33.39
361.23 31.00
116.90 4.97
115.57 7.87
112.97 21.81
115.70 61.19
*-* **
** n.a.
** **
Cu
-mg/kg-
20.99
21.33
19.95
31.16
31.22
24.29
33.18
35.71
17.58
25.48
20.40
17.61
n.a.
**
n.a.
Hi

1.73
1.57
1.30
2.23
1.48
3.01
2.52
2.53
1.29
2. 09
1.71
1.33
*
**
n.a.
Cr

3.75
3.96
3.76
4.46
4.54
4.31
3.29
3.54
2.93
2.28
2.74
2.60
n.a •
**
n.a.
Pb Al

2.05 892.67
2.12 1150.00
1.70 669.00
2.30 1126.00
1.74 681.00
3.88 2132.33
3.18 1226.00
2.42 595.00
1.82 1312.33
1.55 962.00
1.47 1248.33
1.89 1298.67
n.a. n.a.
* n.a.
n.a. n.a.
       *, **=Significantly different at. P<0.05 and P<0.01.  respectively.

       n.a."No significant differences.

-------
       TABLE 47. CONCENTRATIONS OF SELECTED CHEMICAL ELEMENTS (DRY WEIGHT) IN  SPINACH CROWN ON BLOUNT SILT LOAM AMENDED WITH
                 SEWAGE SLUDGE. SLUDGE PLUS CdClj AND CdCl2 AT RATES TO PROVIDE  EQUIVALENT AMOUNTS OF TOTAL SOIL-Cd.  SLUDGE AND
                 SLUDGE PLUS CdCl2 CONTAINED 400 ing/kg (DRV WEIGHT) OF Cd.
oo
0)
Cd -Source Cd-Loading Rate

Sludge
Sludge *
CdCl2
CdClj
kg /ha 	
1.25
2.5
5.0
10.0
1.25
2.5
S.O
10.0
1.2S
2.5
5.0
10.0
K

7.09
5.76
6.25
6.34
6.56
5.69
6.70
6.12
6.86
6.75
5.88
5.01
Na

0.488
0.784
0.553
0.716
0.559
0.533
0.527
0.562
0.574
0.544
0.718
0.636
Ca

1.98
2.47
2.46
1.99
1.94
2.14
1.92
2.67
1.89
1.66
1.93
2.04
Hg

2.10
2.14
2.06
2.26
1.85
2.47
2.38
1.87
1.98
1.93
1.96
2.46
Fe Mn

723.33 128.87
751.00 134.70
787.67 145.93
484.67 104.07
956.33 145.33
853.67 166.97
897 33 171.20
795.67 173.87
739.33 115.17
564.67 103.63
843.33 122.27
799.00 147.77
Zn Cd

177.27 6.45
205.33 8.17
324.70 14.01
469.60 16.22
160.00 16.51
227.10 23.43
242.97 28.60
408.57 47.42
116.90 4.97
115.57 7.87
112.97 21.81
115.70 61.19
Cu

-mg/kg-
14.91
18.63
18.46
22.31
8.14
23.81
23.29
32.72
17.58
25.48
20.40
17.61
Ni

2.55
3.29
2.67
2.20
1.98
2.45
2.42
3.11
1.29
2.09
1.71
1.33
Cr

5.02
6.17
5.17
7.54
5.31
5.66
4.14
5.22
2.93
2.28
2.74
2.60
Pb Al

2.60 907.00
2.21 932.00
1.50 1020.00
1.15 647.33
1.79 1362.33
3.35 1214.00
2.30 1349.00
2.63 1089.33
1.82 1312.33
1.55 962.00
1.47 1248.33
1.89 1298.67
i
F Ratio Significance Levels
Loading Rate
Cd Source
Interaction



n.s.
n.a.
n.s.
n.s
n.e.
n.s.
n.a.
**
*
n.s.
n.s.
n.a.
n.a. n.a.
n.s. **
n.a. n.a.
** **
** **
** **
*
n.s.
n.s.
n.a.
**
n.s.
n.a.
**
n.a.
n.s. n.a.
n.s n.s.
n.a. n.a.
       *,  **-Significantly different at P£0.05 and P<0.01,  reapectively.

       n.a."No significant differences.

-------
TABLE 48.  CONCENTRATIONS OF SELECTED CHEMICAL ELEMENTS (DBY WEIGHT)  IN SPINACH GROWN ON BlflUNT SILT LQAM AMENDED WITH
           SEWAGE  SM'DCE, SLUDGE PLUS CdCl2 AND CdCl2 AT KATES TO PROVIDE EQUIVALENT AMOUNTS OF TOTAL SOIL Cd.  SLUDGE
           AND SLUDGE  PLUS CdCl2 CONTAINED 600 Big/kg (DRY WEIGHT) OF Cd.
Cd-Source
Sludge
Sludge *
CdCl2
CdCl2
Cd-Loading Rate
1.25
2.5
5.0
10.0
1.25
2.5
5.0
10.0
1.25
2.5
5.0
10.0
K

6.65
7.29
5.80
5.84
6.15
6.70
6.37
5.84
6.86
6.75
5.88
5.01
Na

0.526
0.658
0.554
0.737
0.709
0.584
0.488
0.571
0.574
0.544
0.718
0.636
Ca

2.20
2.16
2.55
2.38
1.97
2.02
2.09
2.32
1.89
1.66
1.93
2.04
Mg

1.84
1.88
2.14
2.29
1.60
2.01
2.08
2.07
1.98
1.93
1.96
2.46
Fe

710.00
364.67
1427.67
638.67
538.00
802.67
1347.00
759.00
739.33
564.67
843.33
799.00
Mn

113.20
133.33
164.90
140.20
106.50
127.17
140.43
99.03
115.17
103.63
122.27
147.77
Zn Cd

134.37 6.42
206.77 9.14
243.03 14.00
358.97 20.57
121.57 7.25
146.47 11.70
206.47 27.26
245.27 32.83
116.90 4.97
115.57 7.87
112.97 21.81
115.70 61.19
Cu

-mg/kg—
15.62
17.61
15.56
23.89
22.79
18.12
22.71
22.13
17.58
25.48
20.40
17.61
Hi

1.70
1.46
1.95
1.44
1.25
1.11
1.36
1.32
1.29
2.09
1.71
1.33
Cr

5.97
6.02
5.33
5.65
4.46
4.19
3.27
2.98
2.93
2.28
2.74
2.60
Pb Al

1.93 1029.00
1.93 485.00
3.83 2227.67
1.83 905.33
2.36 821.33
2.24 1115.67
7.03 1861.67
2.11 1030.67
1.82 1312.33
1.55 962.00
1.47 1248.33
1.89 1298.67
F Ratio Significance Levels
Loading
Rate
Cd Source
Interaction
*
a. a.

n.a.
n.a.

n.s.
*

**
a. a.

n.a.
n.a.

n.a.
n.s.

** **
** **
** **
n.a.
n.a.
n.a.
n.a.
n.a.
n.a.
n.a.
**
n.a.
* n.a.
n.a n.a.
n.a. n.a.
*, **=Significantly different at P<0.05 and P<0.01, respectively.

n.a.-No aignificant differences.

-------
       TABLE 49. CONCENTRATIONS OF SELECTED CHEMICAL ELEMENTS (DRV WEIGHT) IN SPINACH  CROWN ON BLOUNT SILT LOAM AMENDED WITH
                 TWO DIFFERENT SLUDGE PLUS CdCl2 MIXTURES OR CdCl2 AT RATES TO PROVIDE EQUIVALENT AMOUNTS  OF TOTAL SOIL-Cd.
                 BOTH SLUDGES CONTAINED  1000 log/kg (DRY WEIGHT) OF Cd.  SLUDGE I WAS  COMPOSED OF SLUDGE WITH 40 rag Cd/kg PLUS
                         SLUDGE n WAS COMPOSED OF SLUDGE WITH 140 mg cd/kg PLUS Cdci2-
o
o
Cd-Source Cd-Loading Rate
	 kg /ha 	
Sludge I 1.25
2.5
5.0
10.0
Sludge II
CdCl2
1.25
2.5
5.0
10.0
1.25
2.5
5.0
10.0
F Ratio Significance Levels
Loading Rate
Cd Source
Interaction
K
7.26
5.93
7.11
6.41
6.67
6.34
5.81
6.72
6.86
6.75
5.88
5.01
n.a.


Ha
Ca
0.533 1.85
0.506 2.06
0.561 1.88
0.570 2.09
0.411
0.524
0.682
0.546
0.574
0.544
0.718
0.636
n.a.


7.61
1.93
1.82
2.03
1.89
1.66
1.93
2.04
n.a.


MR
1.76
1.92
1.99
2.21
2.13
1.92
1.97
2.16
1.98
1.93
1.96
2.46
**


Fe Mn

492.33 70.13
1311.67 122.97
452.67 88.93
1146.00 116.60
503.00 131.00
819.33 116.40
820.00 128.33
460.33 115.33
739.33 115.17
564.67 103.63
843.33 122.27
799.00 147.77
n.a. n.a.


Zn Cd

102.63 9.83
98.20 11.97
1131.80 18.81
169.60 11.76
119.53 2.25
121.83 4.93
14B.73 13.79
169.03 12.12
116.90 4.97
115.57 7.87
112.97 21.81
115.70 61.19
** **
** **
*•* **
Cu

-rag/kg—
16.01
15.80
18.75
18.31
19.17
17.67
20.00
19.17
17.58
25.48
20.40
17.61
n.a.
n.a.'.
n.a.
Hi

1.74
1.73
1.52
1.87
1.43
1.23
1.20
1.41
1.29
2.09
1.71
1.33
n.a.
n.a.
n.a.
Cr

2.78
3.94
3.17
4.08
2.17
3.18
2.89
2.28
2.93
2.28
2.74
2.60
n.a.
**
n.a.
Pb Al

1.88 705.33
5.44 1854.33
1.49 693.00
2.38 1756.00
8.98 825.00
3.31 1206.00
2.47 1337.33
1.51 710.33
1.82 1312.33
1.55 962.00
1.47 1248.33
1.89 1298.67
n.a. n.a.
n.a n.a.
n.a. n.a.
       *,  **-Significantly different at P<0.05 and P<0.01,  reapectively.

       n.a.-No  significant difference!.

-------
   The main objectives of the study reported here were to produce corn (Zea
mays L.) grain and soybean (Glycine max L.) with the highest concentration of
Cd obtainable from healthy plants for use in formulating rations for chicks
and throughout their productive life as layers.  Laying hens consume higher
amounts per body weight of food materials than other animals and would there-
fore be more likely to exhibit adverse health -and performance effects from
the absorption and accumulation of Cd than other animals.

Methods and Materials

   Single crosses of corn were selected on the basis of their capacities to
accumulate high (Nol7 x Frl4A) and intermediate (R802A x R806) concentrations
of Cd in grain from the sewage sludge-amended strip-mined spoil where they
were planted.  Sufficient corn grain having Cd concentrations of 0.71 and
0.35 mg/kg was obtained from the strip-mined area to provide materials for
formulating the high and intermediate Cd levels for the feeding study.  Com-
mercial corn grain containing less than 0.06 mg Cd/kg of grain was used in
low-Cd rations.  After processing, the meal from soybeans (Woodworth and
Harosoy 63) grown on sludge-amended strip-mined spoil contained 2.38 mg/kg
of Cd and was used to formulate the high-Cd rations.  The intermediate-Cd
rations were formulated using a half and half mixture of the high-Cd soybean
meal and a commercial soybean meal containing less than 0.06 mg Cd/kg.  The
commercial meal was used in the low-Cd rations.  Mineral and vitamin sup-
plements were added to rations in amounts and from sources that are generally
used in commercial poultry operations.  The experimental diets are shown in
Table 50.  Mean concentrations of Zn, Cd, Cu, Mn, Fe, Pb, Cr, Ni, Se, Mg, Ca,
and F in low-, medium-, and high-Cd diets (LCd, MCd, and HCd) are presented
in Table 51.

   Three hundred commercial hybrid pullet chicks (Hyline W36) were brooded
in lots of 25 each in a Petersime Brooder Battery.  At six weeks of age they
were transferred by lots to growing batteries.  From 20 to 80 weeks of age
they were housed two birds per cage in 25.4 x 45.7 cm laying cages.  The HCd,
MCd and LCd diets (Table 51)  were each fed to four lots of birds ad_ libitum.
Starter, developer, and layer diets were fed from 0-8, 8-20, and 20-80 weeks
of age, respectively.  Distilled or deionized water was provided ad libitum
in stainless steel or plastic waterers throughout the assay.

   The birds were housed in environmentally controlled quarters maintained
at 16-27 C with heaters and air conditioners as needed.  A standard step
down-step up lighting schedule was provided following the first five days
during which lighting was continuous.

   Feed intake and body weights were determined at biweekly intervals from
0-8 weeks of age and by 4-week periods thereafter.  Egg production was
measured daily and egg weight and specific gravity were determined from a
3-day collection of eggs taken at the end of each 4-week period starting
during the 28th week.

   The experiment was initiated with 4 replications of birds on each of the
LCd, MCd, and HCd diets.  Samples of brids were terminated at 8, 20,  50, 72,
and 80 weeks by cervical dislocation and decapitation.  At 8 and 50 weeks, 4


                                      191

-------
      Table  50. Experimental diet.
vo
ISJ

Ingredient-
High cadmium corn (12. 1Z P)
Medium cadmium corn (10. 6Z P)
Lou cadmium corn (9.6Z P)
High cadmium soybean meal (46. 4Z P)
Lou cadmium soybean meal (51. 4Z P)
Corn gluten meal (60Z P)
Alfalfa meal (17Z P)
DL-methlonlne
Dlcalclum phosphate
Ground limestone
Iodized salt
Manganese sulfate (27Z Mn)
Chollne chloride (50Z)
Vitamin mix 0i/
Layer vitamin mix
Tylosln
Starc^
_
-
69.90
-
23.20
2:00
1.00
.05
2.20
1.00
.40
.05
.05
.10
-
.05
, LBdV
Dev
—
-
82.60
-
13.75
-
1.00
-
1.00
1.00
.40
.05
.05
.10
-
.05
Layer—
_
-
64.50
-
2S.OO
-
1.00
-
1.25
7.50
-.40
.05
.05

.25
~
Starter^
_
56.
13.
11.
11.
2.
1.
g
2.
1.
>
.

f
-
•

59
27
62
62
00
00
05
20
00
40
05
05
10

05
Dev

67
15
6
6

1

1
1






_
.58
.85
.46
.46
-
.00
-
.00
.00
.40
.05
.05
.10
-
.05
Layer—
_
53.70
12.60
11.60
11.60
-
1.00
-
1.25
7.50
.40
.05
.05

.25
"
Starc^
71.63
-
-
21.47
-
2.00
1.00
.05
2.20
1.00
.40
.05
.05
.10
-
.05
, K-V
Dev
86.63
-
-
9.72
-
-
1.00
-
1.00
1.00
.40
.05
.05
.10
-
.05
Layer^
69.10
-
-
20.40
-
-
1.00
-
1.25
7.50
.40
.05
.05

.25
-
      — High,  medium,  low cadmium corn contained .71, .35, and <.06 mg Cd/kg.  High and low cadmium soybean meal contained
        2.38 and <.06  mg Cd/kg.  Dlcalclum phosphate and limestone contained 5.55 and 0.55 mg Cd/kg, respectively.

      -  LCd,  MCd, and HCd starter diets contained .22,  .76, and 1.22 og Cd/kg.

      -1 LCd,  MCd, and HCd layer diets contained .10, .57. and .97 mg Cd/kg.

      4/
      —  Starter, developer and layer diets formulated to contain low (LCd), medium (MCd), and high (HCd) In cadmium.

      -  Provided per  kg of starter and developer diet:  vitamin A 2,000 I.U.; 03 1,000 ICU; K 1 mg; nlacln 27 mg; calcium
         pantathonate  11 mg;  Bj^ 9 meg; blotln 100 meg;  rlboflavln 3.6 mg.  Provided per kg of layer diet: vitamin A
         4400 I.U.;  Dj 1,000 ICU; vitamin K 1.1 tag; nlacln 22 mg; calcium pantothenate 11 mg; B^j 12.5 meg; E 2.2 mg;
         rlboflavln 4.4 mg; chlortetracycllne 20 ing/ton.

-------
Table51-   Mean concentrations of selected elements In feed samples of low-,  medium-,  ami hlgh-Cd
           diets formulated from corn and soybeans grown on sludge-amended soil.
Chemical Element
Cd Dice
Low: x
rt.d.
Medium: x
s.d.
IHijli! x
s.d.
Zn

29.4
3.9
44.4
5.7
48.5
8.9
Cd

0.095
0.047
0.567
0.107
0.966
0.135
Cu

5.44
1.03
5.88
1.49
6.07
1.96
Mu

175
32
170
23
170
32
Fe Pb
7l__.
-——ing/ Kg——— —
393 <0.625
99
398 <0.625
113
380 <0.625
83
Cr

2.23
0.67
2.17
0.36
2.21
0.80
Nl

3.38
0.52
4.53
0.65
4.91
0.88
Se

0.017
0.01
0.025
0.016
0.033
0.012
Mg

0.181
0.016
0.191
0.029
0.155
0.031
Ca

2.54
1 .25
2.44
1.47
2.66
1.36
P

0.8r>7
0.4V,
0.81 |
0.171
0.966
0.263

-------
birds were sampled from each of the 4 replications and tissues collected for
residue and chemical analyses and pathological examination were pooled within
each replication.  Sampling procedures at 20 weeks were the same,  except only
3 birds per replication were sacrificed.  Ac the end of 52 weeks,  2 of the 4
replications of hens fed LCd diets were switched to HCd diets and vice versa.
This created two new treatments designated LCd to HCd and KCd to LCd.   At the
72 and 80 week sampling periods 5 birds from each of the remaining 2 repli-
cations per treatment were randomly selected and tissues from each bird were
analyzed separately.

   Small hematologic blood samples were collected from wing veins and larger
samples by cardiac puncture for serum clinical chemistry analyses.  Measure-
ments of potential manifestation of toxicosis (clinical chemistry, hematology,
and histopathology) were conducted using standard techniques.  Dissection
techniques for collecting samples of heart,  lung, liver, pancreas, spleen,
kidney, breast muscle,  leg muscle, femur bone, brain, crop, proventriculus,
gizzard, and duodenum were previously described in detail (Hinesly et al.,
1976),  along with method preparation and analysis for various chemical con-
stituents.  All tissues and primary wing feathers were analyzed for Zn, Cd,
Cu, Mn, Fe, and Pb.  The shell, white, and yolk of eggs were analyzed for
Cd.  Feathers were washed in double-distilled water, dried (60 C), ground in
a Wiley mill, and digested in concentrated HNO- (90 C) until fuming ceased.
Then they were digested with 30% H-O-, taken to dryness, and dissolved in
0.5 N HNO. for analysis by atomic absorption spectrophotometry.  Eggs were
washed in distilled water before boiling in plastic cooking bags,  after which
shells, white, and yolk were separated, dried (60°C) and ground in a Wiley
mill.  Shells were prepared for analyses by the procedure used for feathers.
Methods used to analyze other chicken tissues were employed for the analysis
of egg whites and yolks.

Results and Discussion

General Health and Performance—

   In the absence of selecting corn hybrids and soybean cultivars for in-
herited capacities to take up high amounts of Cd and growing these on soils
treated with sewage sludges at rates which supply nitrogen and/or phosphorus
in excess of crop requirements, the high-Cd diet (Table 51) represents the
maximum dietary insult  that poultry will encounter from biologically incorpo-
rated Cd.  But there is no indication that enhanced concentrations of Cd ad-
versely affected consumption of feed (Table 52) and cumulative gains in body
weights of chicks, broilers, and hens (Table 53).  Both feed intake and gains
remained fairly constant after 24 weeks, regardless of dietary Cd concen-
trations.  The ratio of feed consumption to body weight gain was not signifi-
cantly affected by Cd treatment.  The rate of egg laying, presented in Table
54, showed no significant differences associated with concentrations of Cd
in diets.  Decreases in egg production with age were about the same for the
three Cd treatments.  The rate of mortality, normally about 1% each 4-week
period, was well below expectations for all diets (Table 55).  Different
concentrations of Cd in diets had no effect on egg weight, shell quality,
and concentrations of Cd in egg parts.  The data presented in Table 56 shows
that egg whites and yolks generally contained Cd concentrations that were
below the detectable limits of 0.062 mg/kg.   Egg shells contained measurable

                                      194

-------
amounts of Cd and chese amounts increased as age of hens increased and rate
of lay decreased, but shell-Cd concentrations were not affected by Cd levels
in diets.

Concentrations of Selected Chemical Elements—

   Concentrations of Zn, Cd, Cu, Mn, Fe, and" Pb in various tissues of 8-week
old chicks are shown in Table 57.  No increased concentration of these metals
in breast and leg muscle could be attributed to higher level in diets.  But,
in gizzard, kidney,  and liver tissues, Cd concentrations reflected levels
of the metal in diets.   Data presented in Table 58 shows that concentrations
of Ca, Na, Mg, K, and P in the several tissues from 8-week old chicks were
not significantly affected by dietary-Cd differences.  The barely signifi-
cant differences in Ca concentrations in breast muscle and kidney tissues
were evidently artifacts as judged by later data.

   Zinc,  Cd, Cu, Mn, Fe, and Pb concentrations in tissues of 20-week old
chickens are presented in Table 59.  Cadmium concentrations in gizzard,  kid-
ney, and liver tissues were significantly increased by increased concentra-
tions in diets.  Furthermore, Cd concentrations in these three tissues from
20-week old birds were significantly higher than those from 8-week old chicks
at all dietary Cd levels.  Other significant differences in metal concentra-
tions in tissues show that these were not related to differences in diets and
in light of data from similar tissues collected at earlier and later dates,
apparently were artifacts.  This is also true for concentrations of Ca,  Ma,
Mg, K, and P in various tissues from 20-week old birds (Table 60).

   In addition to those analyzed at the end of 8 and 20 weeks, several other
tissues collected at 50 and 80 weeks were analyzed for heavy metal concen-
trations and are exhibited in Tables 61 and 64.  At 50 weeks, the concen-
trations in brain and primary wing feathers appeared to decrease as Cd con-
centrations in diets increased, but this was not borne out by samples col-
lected at 80 weeks.   Results from analyses of 50 week samples of breast  muscle,
leg muscle, and femur bone, showed only Cd concentrations to be different in
leg muscle, but unrelated to concentrations of the element in diets.  Con-
centration of Cd in crop, proventriculus, gizzard, gizzard lining, duodenum,
liver, pancreas, kidney, and lung were markedly increased by higher Cd con-
centrations in diets.  The two higher dietary-Cd concentrations may have
caused small, but significant, increased concentrations of Cd in spleen.
Cadmium concentrations in 50 week heart tissue were not affected by dietary-
Cd levels.  Concentrations of Cd in 50 week samples of gizzard, kidney,  and
liver were significantly higher than they were in 20 week samples of the same
tissues at all levels of dietary-Cd.  Concentrations of alkali and alkaline
earth metals and P in 50 week tissue samples were either not significantly
changed or were not related to Cd concentrations in diets (Table 62).

   Tissue samples collected at 72 weeks to facilitate toxicological measure-
ments showed increased concentrations of Cd in gizzard, gizzard lining,  kid-
ney, and liver that corresponded with Cd levels in diets (Table 63).  Whether
or not there were differences in Cd concentrations in breast muscle and  leg
muscle is debatable because statistical analysis was performed by using one-
half of the lowest detectable concentration in each case.  Concentrations of
Cd at 72 weeks were markedly increased over those at 50 weeks only in kidneys.


                                      195

-------
Table  52. Grams of feed consumed per bird per day (g/b/d) during 2- or 4-week
          incerval periods of che 80-week study.

Period
(weeks)
0-2
2-4
4-6
6-3
8-12
12-16
16-20
20-24
24-28
28-32
32-36
36-40
40-44
44-48
48-52

52-56

56-60

60-64

64-68

68-72

72-80

Diet
g/b/d













L-li/
L-H
L-L
L-H
L-L
L-H
L-L
L-H
L-L
L-H
L-L
L-H
L-L
L-H
11.85
24.45
36.75
49.53
52.63
47.08
57.45
66.28
90.43
91.67
104.73
97.32
99.10
95.95
100.35
97.55
99.95
91.00
99.95
87.90
95.80
87.05
94.85
82.20
81.65
84.05
87.70
90.45
r
.*
^
±
=
r
j»
= 1
± 1
*
= 3
= 1
= 1
± 1
= 1
± 1
i 7
z 1
± 4
i 1
i 1
± 2
± 2
± 3
± 6
= 7
- 1
.10
.13
.32
.47
.23
.86
.83
.46
.10
.89
.73
.53
.98
.43
.85
.55
.05
.12
.05
.51
.50
.35
.46
.91
.06
.77
.32
.55
11
24
36
50
53
47
55
64
89
91
97
96
96
93
96

93

95

97

90

86

86

g/b/d
.50 ±
.20 ±
.20 =
.15 =
.03 r
.63 ±
.55 =
.35 =
.30 =
.75 =
.68 =
.28 ±
.85 =
.75 ±
.65 i
-
.90 ±
-
.17 ±
-
.55 ±
-
.73 t
-
.55 i
-
.90 ±
-
.14
.30
.24
.28
.58
.26
.65
1.65
1.70
1.07
3.22
3.91
5.56
2.42
1.99

2.53

4.49

3.42

4.00

2.73

3.23














H-si/
H-L
H-H
H-L
H-H
H-L
H-H
H-L
H-H
H-L
H-H
H-L
H-H
H-L
2/b/d
11.38
25.40
36.80
50.68
56.10
49.60
55.78
64.95
87.95
94.83
92.03
95.90
97.30
94.05
92.60
98.10
90.45
97.45
90.35
99.40
87.85
103.05
39.20
98.05
78.95
93.30
83.95
86.40
=
.k
-
j
-
j.
-k
-
-
-
-
-
-
^
^
r
j.
^
=
=
=
^
=
=
.1
-
-
4.
.13
.22
.24
.66
.39
.45
.26
1.49
1.20
3.04
2.12
1.66
4.97
2.67
1.80
.90
6.16
3.16
4.96
.68
9.40
1.65
2.71
.75
3.26
.20
2.0
8.5

—  Four replications to 52 weeks.  At 52 weeks 2 LCd replications were
   changed to HCd (L-H) and 2 continued on LCd (L-L).   Reciprocal changes
   were also made for HCd cveatnent.  Birds on MCd were not changed.
                                      196

-------
Table 53. Cumulative gains in body veighc of survivors for indicated periods.

Period
0-30 weeks



LCd



gm ± SEM
2
4
6
3
12
16
20
24
28
32
36
40
44
48
52

56

60

64

68

72

80



.










i /
T T *-'
L-L—
L-H
L-L
L-H
L-L
L-H
L-L
L-H
L-L
L-H
L-L
L-H
L-L
L-H
70
211
378
570
879
1056
1298
1364
1385
1400
1430
1438
1442
1456
1471
1469
1465
1439
1470
1432
1479
1446
1489
1411
1431
1410
1416
1445
.50 =
.50 =
.00 =
.25 =
.00 =
.25 =
.00 ±
.50 r
.50 ±
.00 i
.25 =
.50 ±
.00 :
.75 ±
.50 ±
.00 ±
.50 =
.00 ±
.00 ±
.00 r
.50 ±
.50 ±
.50 ±
.00 ±
.00 =
.50 ±
.00 ±
.50 r
1.19
2.10
3.49
3.07
1.22
9.34
10.21
4.66
13.57
16.56
7.11
13.22
14.73
11.23
14.50
6.00
10.50
17.00
9.00
9.00
8.50
9.50
12.50
14.00
4.00
29.50
32.00
12.50


Diet
MCd










gm ± SEM gi
68
20$
371
566
872
1064
1287
1345
1360
1360
1376
' 1379
1392
1411
1407

1411

1424

1438

1428

1400

1438

.25 ±
.00 ±
.75 ±
.50 =
.00 =
.75 ±
.00 ±
.50 =
.25 z
.00 =
.25 ±
.75 ±
.25 ±
.00 r
.00 ±
-
.50 ±
-
.00 ±
-
.50 z
-
.00 r
-
.75 ±
-
.00 ±
-
1.11
1.29
2.78
4-. 09
7.38
9.56
10.35
7.38
17.38
16.90
26.57
33.11
24.36
25.66
25.46

23.10

27.68

23.15

11.10

15.40

40.06














1

H-L
H-H
H-L
H-H
H-L
H-H
H-L
H-H
H-L
H-H
H-L
H-H
H-L
64
201
360
560
885
1088
1304
1342
1342
1339
1361
1367
1364
,1391
•1360
1445
1340
1439
1363
1451
1363
1500
1355
1478
1335
1462
1393
1455

HCd




n = SEM
.75 =
.50 =
.00 =
.50 =
.25 =
.25 ±
.25 ±
.25 ±
.50 =
.75 z
.50 ±
.00 ±
.75 ±
.25 ±
.50 =
.00 ±
.00 ±
.50 ±
.50 ±
.00 ±
.50 z
.00 ±
.50 r
.50 ±
.00 ±
.00 ±
.50 ±
.50 ±
.75
3.50
3.87
2.40
3.20
6.66
12.10
14.53
21.30
22.95
22.80
25.93
27.89
28.76
25.50
49.00
46.00
18.50
30.50
16.00
30.50
23.00
30.50
20.50
38.00
20.00
37.50
3.50

—  Four replications Co 52 weeks.  Ac 52 weeks 2 LCd replications vere
   changed Co HCd (L-H) and 2 concinued on LCd (L-L).  Reciprocal changes
   were also made for HCd treatment.  Birds on MCd were not changed.
                                      197

-------
Table 54. Percent of surviving hens laying an egg per day (HD%) from 20-80
          weeks bv 4-week intervals.

Period
(weeks )
20-24
24-28
28-32
32-36
36-40
40-44
44-48
48-52

52-56

56-60

60-64

64-68

68-72

72-76

76-80

LCd
HD% ± SEM



L-L-'
L-H
L-L
L-H
L-L
L-H
L-L
L-H
L-L
L-H
L-L
L-H
L-L
L-H
L-L
L-H
40.70
84.08
85.03
84.35
75.70
69.85
64.98
r64.85
59.80
67.55
56.55
57.45
64.50
54.35
49.45
57.05
48.60
46.55
47.90
35.75
43.90
36.40
45.10
^
^
4*
+
^
+
-
+
=
+
=
+
J
r
i
r
+
^
+
+
+
+
+
2
2
1
1
2
3

4
5
3
13
6
2
1

5

1
4

2
2
7
.99
.38
.73
.00
.67
.21
.92
.66
.72
.25
.45
.45
.70
.85
.95
.25
.40
.85
.72
.65
.50
.00
.60
MCd
HDZ i
44
80
84
84
79
72
63
65

65

63

4

61

56

43

46

.53
.25
.88
.15
.05
.08
.00
.70

.63

.20

.15

.78

.28

.42

.23

: SEM
= 6.52
± 1.38
t 2.39
= 1.35
± 3.15
±5.00
= 5.66
± 2.97
-
= 1.78
-
= 1.37
-
= .58
-
± 3.25
-
± 2.04
-
± 4.58
-
± 5.82
-
HCd

HD% = SEM




H-L
H-H
H-L
H-H
H-L
H-H
H-L
H-H
H-L
H-H
H-L
H-H
H-L
H-H
H-L
36.35 =
86.43 =
88.78 =
87.43 =
80.03 ±
77.38 z
70.53 =
'70.35 ±
62.15 r
59.50 ±
66.80 =
57.90 =
66.80 =
62.40 :
69.10 ±
58.05 ±
63.90 ±
59.35 ±
60.15 ±
35.55 t
43.75 ±
45.00 ±
42.00 i
4.47
3.13
1.25
1.25
2.17
1.73
3.51
2.46
3.56
11.60
5.10
12.90
7.10
13.20
.90
5.65
4.50
5.45
4.85
2.75
.85
4.31
10.00

—  Four replications to 52 weeks.  At 52 weeks 2 LCd replications were
   changed to HCd  (L-H) and 2 continued on LCd (L-L).  Reciprocal changes
   were also made  for HCd treatment.  Birds on MCd were not changed.
                                      198

-------
Table  55. Disposition of birds, 0-30 weeks of age.

Renlicate
1


2


3


4


1-4



Fate!/
Started
Died
To Analysis
Started
Died
To Analysis
Started
Died
To Analysis
Started
Died
To Analysis
Started
Dead
To Analysis
Live, 80 weeks

LCd
26
0
18
25
2
17
25
2
18
25
2
18
101
6
71
24
Diet
MCd
26
4
16
26
1
13
25
3
14
25
2
15
102
10
58
34

HCd
26
1
17
25
2
17
25
1
18
25
2
18
101
6
70
25

—Birds sacrificed at 3, 20, 50, and 72 weeks for  tissue analysis.
                                      199

-------
Table  56.  Mean Cd concentrations in egg constituents sampled at various
           intervals during a 54-week period commencing when egg laying
           began.
Number of Weeks Cd
After Lav ins Began Treatment
6 low
medium
high
F-test
15 low
medium
high
F-cest
24 low
medium
high
F-tesc
32 low
medium
high
F-test
41 low
medium
high
F-test
54 low
medium
high
F-test
White
<0.062
<0.062
<0.062
n.s.
<0.062
<0.062
<0.062
n.s.
<0.062
<0.062
<0.062
n.s.
<0.062
<0.062
<0.062
n.s.
0.317
0.144
0.125
n.s.
<0.062
<0.062
<0.062
n.s.
Yolk
<0.062
<0.062
<0.062
n.s.
0.063
<0.062
0.111
n.s.
<0.062
<0.062
<0.062
n.s.
<0.062
<0.062
<0.062
n.s.
<0.062
<0.062
<0.062
n.s.
<0.062
<0.062
<0.062
n.s.
Shell
0.114
0.149
0.148
n.s.
0.068
0.071
0.090
n.s.
0.101
0.124
0.083
n.s.
0.352
0.365
0.294
n.s.
0.456
0.320
0.340
n.s.
0.303
0.325
0.347
n.s.
                                    200

-------
Table 57.  Concentrations of  selected  elements  in various tissues of hens
           fed low-, aedium-, and high-cadmium  diets  formulated from corn
           and soybeans grown on sludge-amended soil.  Samples taken 3
           weeks after hatching.
Diet

Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD

Zn


19.7
20.2
20.7
n.s.

105
111
106
n.s.

92.4
107
110
8.6**

81.3
80.3
83.7
n.s.

100
103
103
n.s.

Cd


<0.062
<0.062
<0.062 '
n. s.

0.716
3.19
4.54
1.23**

0.304
1.76
3.21
0.813**

0.078
<0.062
<0.062
n.s.

0.176
0.746
1.17
0.226**
Chemical Element
Cu Mn
'kg fat-free dry we
Breast Muscle
1.97 0.198
1.66 0.294
1.61 0.227
n.s. n. s.
Gizzard
8.63 9.82
8.06 10.3
7.80 9.20
0.55** n.s.
Kidney
11.2 13.0
12.4 13.1
12.8 13.4
n. s . n. s .
Leg Muscle
5.03 0.750
4.32 0.510
4.69 0.680
n.s. n.s.
Liver
19.4 19.3
19.2 18.3
18.2 16.4
n.s. 2.1*

7e


6.25
8.80
8.48
n.s.

103
94.0
91.5
7.6*

236
229
241
n.s.

29.5
23.5
29.5
n.s.

444
442
419
n.s.

?b


<0.625
0.640
1.02
n.s.

0.692
<0.625
0.738
n.s.

<0.625
0.970
<0.625
n.s.

0.780
<0.625
1.08
0.192**

<0.625
<0.625
n.s.
*,** = significant at 0.05 and 0.01, respectively.
n.s. = non-significant F-test.
                                      201

-------
Table 58.  Concentrations of selected elements in various tissues of hens
           fed low-, medium-, and high-cadmium diets fomulated from corn
           and soybeans grown on sludge-amended soil.  Samples taken 8
           weeks after hatching.
Diet

Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD
*t ** =
n.s. =

Ca


0.022
0.026
0.059
0.028*

0.053
0.057
o.osr
n.s.

0.039
0.052
0.043
0.009*

0.024
0.020
0.020
n.s.

0.017
0.018
0.017
n.s.
significant at
non-signif leant

Na
37

0.150
0.137
0.143
n. s.

0.373
0.383
0.383
n. s.

0.722
0.741
0.809
n.s.

0.310
0.290
0.323
n.s.

0.361
0.362
0.368
n.s.
0.05 and 0
: F-cest.
Chemical Element
Ms
fat-free dry weis
Breast Muscle
0.136
0.138
0.132
n.s.
Gizzard
0.074
0.077
0.072
n.s.
Kidney
0.098
0.107
0.107
n.s.
Leg Muscle
0.122
0.121
0.125
n.s.
Liver
0.100
0.104
0.098
n. s.
.01, respectively

K

jnc-— —
1.63
1.63
1.59
n.s.

1.38
1.39
1.45
n.s.
•
1.07
1.09
1.20
n.s.

1.76
1.71
1.80
n.s.

1.27
1.31
1.26
n.s.


P


0.940
0.872
0.951
n.s.

0.563
0.596
0.574
n.s.

1.13 -
1.14
1.12
n.s.

0.776
0.770
0.798
n.s.

1.13
1.21
1.06
n.s.

                                     202

-------
Table 59 .  Concentrations of selected elements in various tissues of hens
           fed low-, medium-, and high-cadmium diets formulated from corn
           and soybeans grown on sludge-amended soil.   Samples taken 20
           weeks after hatching.
Diet


Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD

Zn



24.2
19.8
16. 4
n.s.

108
114
116
n.s.

109
116
128
14**

73.6
83.3
75.2
n.s.

119
115
113
n.s.

Cd
•^^^KM^Mt^^mo /
-— — — TI-.— a§/

<0.062
0.0915
0.109
n. s.

0.872
4.69
6.90
1.14**

0.986
7.64
13.6
1.82-**

0.0928
0.0738
0.121
n.s.

0.352
2.17
3.55
0.776**
Chemical Element
Cu Mn
kg fac-free dry wed
Breast Muscle
1.60 0.605
1.98 0.318
1.90 0.762
n.s. n.s.
Gizzard
9.08 14.3
10.5 12.7
8.48 12.8
1.36* n.s.
Kidney
14.0 15.7
15.0 13.2
15.4 14.6
0.9** n.s.
Leg Muscle
7.78 0.902
4.58 1.14
4.15 0.990
n.s. n.s.
Liver
17.9 17.8
17.0 16.1
17.0 16.5
n.s. n.s.

Fe



15.6
14.7
15.3
0.6*

178
183
184
n.s.

465
505
468
n.s.

44.8
51.4
52.3
6.2*

788
749
667
n.s.

?b



<0.625
<0.625
<0.625
n.s.

<0.625
<0.625
<0.625
n.s.

<0.625
<0.625
<0.625
n.s.

1.17
<0.625
<0.625
0.584**

0.848
<0.625
<0.625
n.s .
*, ** = significant at 0.05 and 0.01, respectively.
n.s. = non-significant F-test.
                                    203

-------
Table 60.  Concentrations of seleccad elements in various tissues of hens
           fed low-, medium-, and high-cadmium diets formulated from corn
           and soybeans grown on sludge-amended soil.  Samples taken 20
           weeks after hatching.
Chemical Element
Diet

Ca


Na


Ms
fat-free dry
Z.
___ j _ V*

o


Breast Muscle
Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD
0.008
0.014
0.011
n. s.

0.050
0.040
0.049
n.s.

0.032
0.041
0.035
n.s.

0.024
0.018
0.020
n.s.

0.023
0.031
0.023
n.s.
0.149
0.167
0.154
n.s.

0.355
0.351
0.356
n.s.

0.806
0.770
0.831
n.s.

0.309
0.316
0.303
n.s.

0.347
0.337
0.335
n.s.
0.120
0.123
0.118
n.s.
Gizzard
0.069
0.070
0.067
n.s.
Kidney
0.104
0.100
0.092
0.008**
Leg Muscle
0.109
0.111
0.106
n.s.
Liver
0.101
0.101
0.088
n.s.
1.42
1.32
1.36
n.s.

1.13
1.25
1.21
n.s.

1.32
1.21
1.17
0.096*

1.44
1.37
1.42
n.s.

1.30
1.18
1.16
n.s.
0.915
0.382
0.900
n.s.

0.575
0.589
0.616
n.s.

1.40
1.36
1.35
n.s.

0.892
0.877
0.882
n.s.

1.35
1.37
1.24
n.s.
*,** = significant at 0.05 and 0.01, respectively.
n.s. = non-significant F-test.

                                     204

-------
Table 61.   Concentrations of selected elements in various tissues of hens
           fed low-, medium-, and high-cadmium diets formulated from corn
           and soybeans grown on sludge-amended soil.  Samples taken 50
           weeks after hatching.
Chemical Element
Diet
•
Low
Medium
High
LSD



50.
50.
51.
n.
Zn


9
0
8
s



<0
<0
0
n
Cd


.062
.062
.070
.s.

'
-------
Table 61. Concentrations of selected elements in various tissues of hens
Continued fed low-, nediua-, and high-cadaiua diets foraulated fraa com
and soybeans grown on sludge- an end ad soil. Saaples taken 50
weeks after hatching.
Diet

Zn

Cd
Chemical Elaaer.:
Cu Mr.

?e

?b
	 	 	 mg/kg fat-free dry weight 	 • • 	
Feaur Bone
Low
Mediua
High
LSD

Low
Mediua
High
LSD

Low
Mediua
High
LSD

Low
Mediua
High
LSD

Low
Mediua
High
LSD
229
256
272
n.s.

128
125
132
n.s.

22.5
24.7
29.7
n.s.

107
103
98.9
n.s.

118
125
142
n.s.
<0.062
<0.062
0.0735
n.s.

1.03
4.27
9.34
3.33**

1.39
8.78
13.6
5.79**

<0.062
<0.062
0.188
n.s.

4.61
21.8
36.3
4.3**
0.985 19.0
. 0.743 16.0
0.777 17.0
n.s. n. s.
Gizzard
4.55 3.34
4.80 3.40
5.42 3.78
n.s. n.s.
Gizzard Lining
33.4 21.7
35.8 18.5
56.0 28.2
13.2* n.s.
Heart
16.2 2.30
16.0 2.60
16.0 2.75
n.s. n.s.
Kidney
12.9 10.1
13.1 11.6
13.3 13.2
n.s. 2.4**
135
115
141
n.s.

136
181
183
n.s.

85.8
93.9
74.6
n.s.

240
217
234
n.s.

359
382
330
n.s.
<0.625
<0.625
<0.525
n.s.

0.355
<0.625
0.653
n.s.

1.12
1.45
1.73
n.s.

0.798
<0.625
<0.625
n.s.

<0.625
<0.525
<0.625
n.s.
*,•**= significant at 0.05 and 0.01, respectively.
n-s. = non-significant F-cest.
                                    206

-------
Table 61 . Cone ant rations of selected elements in various tissues of hens
Continued fed low-, medium-, and high-cadmiua diets formulated from com
and soybeans grown on sludge-amended soil. Samples taken 50
weeks after hatching.
Chemical Element
Diet

Zn


Cd


Cu
Mn
kg fat-free dry vei
Fe


?b


Leg Muscle
Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD
116
118
108
n.s.

123
138
134
n.s.

48.4
51.6
49.4
n.s.
0.111
0.163
0.110
0.039*

0.898
3.57
6.18
0.190**

<0.062
0.128
0.319
0.140**
4.05
3.70
4.06
n.s.

16.0
14.7
16.2
n.s.

2.19
2.29
2.43
n.s.
0.874
0.762
0.760
n.s.
Liver
9.40
13.0
11.9
n.s.
Lung
1.96
2.20
2.23
n.s.
81.0
64.0
62.0
n. s.

337
303
257
n.s.

771
809
780
n. s.
0.336
<0.525
<0.525
n.s.

0.998
<0.625
<0.625
n.s.

<0.625
<0.625
<0.625
n.s.
Pancreas
Low
Medium
High
LSD
115
127
129
n.s.
0.299
1.85
2.90
0.586**
6.85
6.15
6.01
n.s.
13.0
13.5
14.0
n.s.
182
164
142
n.s.
<1.00
<1.00

-------
   Table  til.   Cor.centrai.onr,  :f  •:ciacred aienencs lr. various cissues of her.s
   Cor.cir.ued   fee:  low-, ae-iiu=s-, c.nd .-i.^h-cada^ua dz.eta for=uiacec. froa corn
              ar.ri  joyteanj grown cr. sludsc-anended soil.  S-znples taken 50
Diet


Zr.


Cd
	 -,/
Cha.-i£il
Cu
.'.g :az-i.r
. -ilasanc
Mn

7s

?b
•ee dry weight 	
Spleen
Low
Xadiun
High
LSD
33.?.
S6.S
56. 3
3.1*
0.349
1.53
!..56
0.931*
4.56
•i . /6
4.62
n.s.
2.25
2.27
2.03
n.s.
892
314
841
n.s.
<1.00
<1.00
C1.00
n.s.
      ** - sicniiicant ar 0.05 and 0.01, respectively.
      . =• non-sign:ficar: ?-r.ssc.
Ic is noceworchy that for hens switched from low- to high-Cd diets, tissues
that were notable accumulators of Cd gained as much of the metal as was lost
by the same tissues in hens that were switched from high- to low-Cd diets.
Changes in concentrations of other transition or heavy metals were either
not related to Cd levels in diets or were not supported by concentration
changes of these metals in similar tissues collected earlier and later than
72 weeks.

   As can be seen in Table 64, concentrations of Cd were significantly in-
creased in 80 weeks crop, proventriculus,  muscular gizzard, gizzard lining,
and duodenum tissues.  Furthermore, relatively large changes in concentra-
tions occurred when hens were1 switching from one dietary-Cd level to another.
At 80 weeks, enhanced concentrations of Cd in liver, kidney, pancreas, and
spleen tissues paralleled levels of the metal in diets, although differences
in concentrations of the latter tissues were small in comparison to changes
in kidney.  It was unlikely that concentration of Cd in heart and lungs was
affected by diets, because difference in concentration was dependent on the
use of one-half the lowest detectable level to show significance.  The same
is true for leg muscle tissue.  No differences were found in breast muscle.
A small, but significant enhancement of Cd concentrations in femur bone with
higher dietary-Cd levels was found at 80 weeks.  Concentrations of Cd in
brain and feathers were unaffected by dietary-Cd levels, although an insig-
nificant trend for higher levels in feathers was observed.  Concentrations
of Cd were significantly changed in 80 week liver, kidney, pancreas, and
spleen tissues when hens were switched to different diets, but only liver
and kidney tissues had markedly higher or lower Cd concentrations as a result
of reciprocal changes in dietary-Cd levels.

   In a comparison of Cd concentrations in tissues of 50- and 80-wee< old
hens, enhanced levels with age occurred only in kidney tissues.  During the
30-week interval, Cd concentrations in kidney were increased a little less


                                      208

-------
Table 62.  Concentrations of selected elements in various tissues of hens
           fed low-, nedium-, and high-cadmium diets formulated from com
           and soybeans grown on sludge-amended soil.  Samples taken 50
           weeks after hatching.
Chemical Element
Diet


Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD

Low
Medium
High
LSD
Ca



0.0175
0.0126
0.0113
n.s.

0.0791
0.0527
0.0323
n.s.

0.183
0.116
0.178
n.s.

0.0646
0.0612
0.0503
n.s.

0.0248
0.0240
0.0246
n.s.
Na
. ,_.,.,. m?


0.176
0.173
0.187
U.S.

0.512
0.436
0.447
0.052*

0.0894
0.0753
0.0855
n.s.

0.835
0.992
0.941
n. s.

0.378
0.340
0.364
n. s.
Mz
fac-free dry we:
Breast Muscle
C.112
0.114
0.112
n.s.
Gizzard
0.0663
0.0663
0.0778
n.s.
Gizzard Lining
0.0113
0.0099
0.0160
n.s.
Kidney
0.0908
0.0911
0.0859
n.s.
Leg Muscle
0.0975
0.0971
0.101
n.s.
X
f -rnr •-<> -
Lg ut — • ••»

1.28
1.44
1.43
n.s.

1.46
1.38
1.51
n.s.

0.120
0.120
0.129
n.s.

1.37
1.40
1.30
n.s.

1.53
1.46
1.45
n.s.
p



0.717
0.810
0.790
n.s.

0.430
0.379
0.421
n.s.

0.277 '
0.322
0.306
n.s.

1.25
1.14
1.21
n.s.

0.813
0.670
0.730
n.s.
*, ** = significant at 0.05 and 0.01,  respectively.
n.s.  = non-significant F-test.
                                     209

-------
       ei 62.   Concentrations of  selected elements  in various  tissues of hens
       ins:cd   fed low-,  neci^-,  ar.d hlsh-cadsiir:  dials  foraulate:'. fran corn
              and soybeans  sro'--n ou siur'~u-anar.ded soil.  Samples ta'/.^i 30
              veeks afrar  iiacch^ng.

                                    Cheaical -Z
   Diet            Ca         X-i
                               	\ fat-fre».  dry  veig:

                                         Liver
Low
Medium
Kign
LSD
0.0212
0.0293
0.023-4
n.s.
0.393
0.345
0.345
r..s.
0.0363
0.0874
0.0323
n. s.
1.20
1.32
1.31
n. s.
1.14
1.12
1.05
n.s.
                --p.t  ac 0.50 ar.d O.C1, respectively.
   r.s.  =  non-significant F-test.


Chan two-fold, regardless of the concentration of  Cd in the diet.   In other
tissues that accumulated Cd, concentrations appeared to have reached an
equilibrium with dietary levels of the metal at 50 weeks of age and thus,
remained unchanged with time.  For hens that were  switched from high- to
low-Cd diets at 50 weeks, only kidney tissues contained higher concentra-
tions of Cd 30 weeks later.

Clinical Parameters—
   There was some indication of low packed cell volumes (PCVs) in  the high
Cd group  (Table 65), but all of the individual data will require collective
evaluation.  Variations in mean corpuscular volume (MCV) were frequently more
pronounced within treatment group, with distinctly different populations,
than among group means (Table 66).  It appeared that the high Cd group may
have a distorted neutroph.il/lymphocyte ratio at 8 weeks, but it later became
evident that this parameter, too, was quite variable within treatment groups
(Table 67).  Counting avian leukocytes is an extremely arduous and  time-
consuming  task, so this procedure was discontinued in the 70 and 80 week
birds because of the lack of clear patterns.  Serum chemistries (Table 68)
also failed to show consistent trends.  There was, however, a slight ten-
dency for  the high dose group to contain a greater frequency of birds with
"low" serum calcium; this could conceivably be related to Cd interference
with Ca metabolism, but the test employed unfortunately had a limited range.

Relative Organ Weights—

   The liver and kidney tended to be slightly larger in the higher Cd groups,
but the tendency was rather weak and inconsistent (Table 69).  The gizzard
did accumulate high concentrations of Cd early in the study.   Anke et al.
(1970) reported epithelial changes in Gizzards from Cd treated chickens which
were reminiscent of Zn deficiency.  In an acutely Cd-poisoned chick we found
the gizzard was visibly enlarged, so this organ may be particularly susceptible
to Cd.  Nevertheless, by 72 weeks the differences disappeared, indicating that

                                      210

-------
Table 63 .   Concentrations of selected elements in various tissues of hens
           fed low-,  medium-,  and high-cadmium diets formulated from corn
           and soybeans grown on sludge-amended soil.   After 50 weeks,
           subgroups  from those hens being fed the low- and high-cadmium
           diets were switched to high- and low-cadmium diets,  respectively.
           Samples taken 72 weeks after hatching.
Chemical Element
Diet

Zn

Cd

	 mg/kg
Cu
Mn
Fe
Pb
fat-free dry weight 	
Breast Muscle
Low
Medium
High
Low-High
High-Low
LSD

Low
Medium
High
Low-High
High-Low
LSD
19
19
19
19
18
n

134
130
131
132
118
.8
.9
.9
.7
.9
. s.






9**
0
0
0
<0
0
0

1
6
11
7
3
2
.063
.076
.067
.062
.075
.021**

.06
.77
.2
.54
.87
.05**
1
1
1
1
1
n

5
5
5
5
4
n
.50
.41
.51
.53
.37
.s.

.71
.10
.80
.54
.80
.s.
0
0
0
0
0
0
Gizzard
4
4
4
3
4
n
.607
.551
.460
.437
.531
.112*

.68
.25
.40
.76
.69
. s.
15
15
15
13
16
n

226
212
220
223
197
n
.7
.2
.7
.8
.3
.s.






.s.
<0.625
<0.625
<0.625
<0.625
<0.625
n.s.

<1.00
<1.00
<1.00
<1.00
<1.00
n.s.
Gizzard Lining
Low
Medium
High
Low-High
High-Low
LSD

Low
Medium
High
Low-High
High-Low
LSD
* ft* =
16
22
28
24
18
6

124
142
150
135
134
n
.7
.3
.5
.0
.9
.7*






.s.
significant
n.s. = non-signif icj
1
8
14
12
3
2

8
42
64
19
44
15
at 0
.36
.04
.3
.0
.98
.97**

.32
.2
.4
.5
.4
.8**
.05 and
39
36
38
38
36
n

13
14
16
23
14
n
0.
.0
.6
.7
.3
.6
. s.

.6
.6
.1
.7
.9
.s.
01,
21
23
22
22
30
n
Kidney
13
12
12
11
13
n
.4
.8
.5
.4
.5
.s.

.0
.0
.3
.8
.1
.s.
97
104
93
79
84
n

444
436
412
423
433
n
.0

.4
.8
.9
.s.






.s.
<1.00
<1.00
1.89
1.17
<1.00
0.792*

<1.00
<1.00
<1.00
<1.00
<1.00
n.s.
respectively.
mt F-test.
                                     211

-------
Table 6 3 .
Continued
Concentrations of selected elements in various tissues of hens
fed low-, medium-, and high-cadmium diets formulated from corn
and soybeans grown on sludge-anended soil. After 50 weeks,
subgroups from those hens being fed the low- and high-cadaium
diets were switched to high- and low-cadmium diets, respectively.
Samples taken 72 weeks after hatching.
Chemical Element
Diet


Low
Medium
High
Low-High
High-Low
LSD

Low
Medium
High
Low-High
High-Low
LSD
Zn



114
110
104
113
106
n. s.

119
150
147
139
109
28*
Cd

— — — • — mg

<0.062
0.111
0.136
0.081
0.124
0.037**

1.48
4.06
5.81
2.50
4.49
1.80**
Cu
Mn
/kg fat-free dry we
Leg
3.77
3.85
3.27
3.90
0.313
0.712**

15.7
16.8
17.0
15.5
13.9
n.s.
Muscle
1.08
0.880
1.04
0.815
0.834
n.s.
Liver
11.7
9.48
10.3
11.8
9.15
n.s.
Fe



62.4
59.3
55.9
69.4
64.2
n.s.

397
374
452
573
305
n.s.
Pb



<0.625
<0.625
<0.625
<0.625
<0.625
n. s .

<1.00
<1.00
<1.00
<1.00
<1.00
n.s.
*, ** = significant at 0.05 and 0.01, respectively.
n.s. = non-significant F-test.
                                     212

-------
Table 64.   Concentrations of selected elements in various tissues of hens
           fed low-, medium-, and high-cadmium diets formulated from corn
           and soybeans grown on sludge-amended soil.  After SO weeks,
           subgroups from those hens being fed the low- and high-cadmium
           diets were switched to high- and low-cadmium diets,  respectively.
           Samples taken 80 weeks after hatching.
Chemical Element
Diet


Low
Medium
High
Low-High
High-Low
LSD
Zn



50.8
48.3
54.7
50.1
52.4
n.s.
Cd
	 mo/'
—————nig/

0.101
<0.092
<0.092
<0.092
<0.092
n.s.
Cu
Mn
Fe
?b
kg fat-free dry weight 	

12.2
11.7
12.2
11.4
12.3
n.s.
Brain
1.60
1.51
1.66
1.70
1.62
n.s.

109
105
112
103
109
n.s.

2.05
1.77
1.93
1.72
1.89
n.s.
Breast Muscle
Low
Medium
High
Low-High
High-Low
LSD

Low
Medium
High
Low-High
High-Low
LSD
19.8
18.4
19.3
18.4
19.0
n.s.

79.9
77.8
86.9
73.0
81.6
8.2*
<0.062
<0.062
<0.062
<0.062
<0.062
n.s.

0.243
0.400
0.528
0.294
0.458
0.078**
1.76
1.71
1.64
1.44
1.63
n.s.

5.18
5.91
5.84
4.36
5.79
n.s.
0.351
0.316
0.263
0.381
0.427
0.109**
Crop
1.82
4.60
4.43
3.93
3.30
n.s.
17.8
16.2
14.4
14.0
16.6
n.s.

63.5
74.8
75.3
62.1
82.6
n.s.
<0.625
<0.625
<0.625
<0.625
<0.625
n.s.

<0.830
0.922
<0.830
<0.830
<0.830
0.313**
Duodenum
Low
Medium
High
Low-High
High-Low
LSD
92.2
94.4
106
99.7
94.4
n. s.
*, ** = significant at
n.s. = non-!
significant
0.523
1.23
1.98
1.33
0.612
0.646**
0.05 and 0.
F-test.
7.48
7.20
8.13
7.55
7.50
n.s.
5.66
5.28
5.52
5.90
6.05
n.s.
213
184
279
209
250
n.s.
<1.00
1.05
1.13
2.03
<1.00
n.s.
01, respectively.




                                    213

-------
Table 64. Concentrations of selected elements in various tissues of hens
Continued fed low-, medium-, and high-cadmium diets formulated from corn
and soybeans grown on sludge-amended soil. After 50 weeks,
subgroups from those hens .being fed the low- and high-cadmium
diets were switched to high- and low-cadmium diets, respectively.
Samples taken 80 weeks after hatching.

Diet

Zn

Cd
Chemical Element
Cu Mn

Fe

Pb
	 mg/kg fat-free dry weight 	

Low
Medium
High
Low-High
High-Low
LSD

Low
Medium
High
Low-High
High-Low
LSD

Low
Medium
High
Low-High
High-Low
LSD

Low
Medium
High
Low-High
High-Low
LSD
* ** =

153
190
219
188
235
n.s.

206
233
268
257
246
n.s.

109
117
116
120
107
n.s.

15.5
17.5
18.4
20.6
16.0
n.s.
significant at
n.s. = non-significant

0.073
0.100
0.155
0.118
0.108
n. s.

0.145
0.152
0.221
0.162
0.176
0.046*

1.11
6.06
9.96
9.00
3.22
2.33**

2.08
9.14
15.0
14.9
3.96
2.82**
0.05 and
F-test.
Feathers
6.17 1.60
6.51 1.56
6.58 2.03
6.92 1.55
6.99 3.42
n.s. 1.50**
Femur Bone
1.17 9.74
1.10 8.44
1.08 8.89
0.950 8.16
1.24 8.29
n.s. n.s.
Gizzard
4.72 4.13
5.18 3.88
5.02 3.46
6.25 4.04
4.32 4.07
n. s. n.s.
Gizzard Lining
38.2 11.0
37.5 13.2
34.9 10.9
32.8 15.6
33.3 12.9
n.s. n.s.
0.01, respectively.


41.3
41.4
39.5
36.1
44.4
n. s.

84.4
73.1
87.3
66.7
79.8
n.s.

167
204
213
184
196
30**

79.7
82.4
88.9
94.7
81.5
n. s.



4.44
6.65
4.87
4.04
5.04
n.s .

1.49
1.58
2.15
1.70
1.79
0.41*

1.19
<1.00
1.08
<1.00
1.03
n.s.

1.05
<1.00
1.63
1.02
1.06
n. s.


214

-------
Table 64.
Continued

Diet
Concentrations of selected elements in various tissues of hens
fed low-, medium-, and high-cadmium diets formulated from corn
and soybeans grown on sludge-amended soil. After SO weeks,
subgroups from those hens being fed the low- and high-cadmium
diets were switched to high- and low-cadmium diets, respectively.
Samples taken 80 weeks after hatching.

Zn

Cd
Chemical Element
Cu Mn
	 mg/kg fat-free dry wei

Low
Medium
High
Low-High
High-Low
LSD

Low
Medium
High
Low-High
High-Low
LSD

Low
Medium
High
Low-High
High-Low
LSD

Low
Medium
High
Low-High
High- Low
LSD

101
104
105
108
107
n.s.

124
144
155
133
149
21**

103
105
96.4
105
106
n.s.

142
179
177
179
146
n. s.

<0.077
<0.077
0.114
<0.077
0.090
0.051**

9.52
45.8
69.5
21.8
51.3
9.57**

<0.062
0.113
0.131
<0.062
0.115
0.045**

1.43
4.86
6.66
3.93
4.20
1.64**
Heart
15.4 2.35
15.5 2.44
16.3 2.45
16.4 2.60
15.8 2.55
n.s. n.s.
Kidney
14.3 12.7
15.0 12.3
14.6 12.2
13.8 11.3
14.3 11.5
n.s. n.s.
Leg Muscle
4.02 0.767
3.77 0.727
3.42 0.757
3.41 0.693
3.52 0.733
n.s. n.s.
Liver
18.8 16.3
19.1 13.8
18.7 15.5
19.2 17.0
17.3 14.3
n.s. n.s.

Fe
fTVit" —__«._—
gnc 	

217
227
243
225
235
n.s.

281
264
234
269
288
n.s.

68.1
68.2
63.4
61.3
70.3
n.s.

740
567
500
585
665
n. s.

Pb



<0.775
<0.775
<0.775
<0.775
<0.775
n.s.

<1.00
<1.00
<1.00
<1.00
<1.00
n.s.

<0.625
<0.625
<0.625
<0.625
<0.625
n.s.

<1.00
<1.00
<1.00
<1.00
<1.00
n.s.
*, ** = significant at 0.05 and 0.01,  respectively.
n.s. = non-significant F-test.

                                    215

-------
Table 64.
Continued
Concentrations of selected elements in various tissues of hens
fed low-, medium-, and high-cadmium diets formulated from corn
and soybeans grown on sludge-amended soil. Afcer 50 weeks,
subgroups from chose
hens being fed the low- and
high-cadmium
diets were switched to high- and low-cadmium diets, respectively.


Diet


Low
Medium
High
Low-High
High-Low
LSD

Low
Medium
High
Low-High
High-Low
LSD
"
Low
Medium
High
Low-High
High-Low
LSD

Low
Medium
High
Low-High
High-Low
LSD
Samples

Zn
	 	 	

63.9
58.8
61.2
65.2
54.3
n.s.

112
111
144
121
112
n.s.

70.3
70.8
74.6
75.4
76.2
n.s.

83.4
85.9
84.3
90.5
85.0
n.s.
*, ** = significant
n.s. = non-
sign if ic,
taken 30 weeks after hatching.

Cd
	 mg/

<0.098
0.149
0.277
0.126
0.234
0.154**

0.373
1.86
2.78
1.11
2.11
0.609**

0.289
1.16
1.83
0.853
1.30
0.391**

0.358
1.19
1.49
0.971
1.41
0.463**
at 0.05 and
ant F-test.
Chemical Element
Cu Mn Fe
kg fat-free dry weight 	
Lung
2.16 1.13 875
1.84 1.03 1060
2.20 0.855 1350
1.94 1.25 1200
2.37 1.18 868
n.s. n.s. 303**
Pancreas
4.18 6.49 147
4.14 6.69 93.8
4.33 6.91 162
4.20 8.49 164
3.69 7.93 138
n.s. n.s. n.s.
Proventriculus
14.3 10.5 108
18.8 10.6 96.2
19.5 10.2 108
24.9 11.2 144
13.4 11.1 125
6.9** n.s. n.s.
Spleen
5.08 1.54 867
6.16 1.52 935
8.84 1.56 874
9.86 1.72 982
6.84 1.67 941
3.41** n.s. n.s.
0.01, respectively.


Pb
_..«__«•_«—••—•—

<1.34
<1.34
<1.34
1.53
<1.34
n. s .

<0.954
<0.954
<0.954
<0.954
<0.954
n.s.

<0.661
<0.661
1.20
<0.661
<0.661
n.s.

<2.64
<2.64
4.30
<2.64
<2.64
n.s.


216

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Table 65.  Hematological parameters  in  white leghorn hens having received
          sludge-fertilized corn-soybean diets  from one week of age.

Age and
Relative              	
Dietary Cd (n)	PCV1^	RBC2)

8 Weeks

Low      (16)          32.3 ±  1.5
                       (29.5-34.5)

Medium   (15)          32.0 r  2.0
                       (29-37)

High     (13)          31.4 i  1.7
                       (28.5-34.5)

20 Weeks

Low       (8)          32.4 ±  2.0
                       (31-36)

Medium    (8)          31.1 ±  2.9
                       (27-35)

High      (8)          32.5 ±  4.1
                       (27.5-39)

50 Weeks
Low       (9)          33.1 ± 3.4           2.1 t  0.35          8.5 r 1.1
                       (28-39)              (1.82-2.93)          (7.0-10.4)
          (1)             26                   2.04                7.4

Medium   (10)          29.9 ± 4.0           2.17 ± 0.31          8.3 ± 1.4
                       (25-37.5)            (1.72-2.67)          (7.0-11.5)

High      (9)          28.0 i 3.4           2.02 ± 0.29          8.2 t 0.6
                       (22-32.5)            (1.65-2.66)          (7.6-9.5)
70 Weeks

Low      (10)          31.2 ± 3.5           2.46 ± 0.45          9.6 ± 1.4
                       (26-36)              (2.13-3.48)          (7.6-11.8)

          (9)          27.4 i 3.5           2.19 ± 0.25          8.5 ± 0.9
                       (22-33)              (1.70-2.59)          (7.0-9.6)
          (1)4)           20.5                 1.44                5.8

                                  (continued)
                                      217

-------
Table 65.  (continued)

Age and
Relative
Dietarv Cd (n)
70 Weeks
Medium
High
H-"L
80 Weeks
Low
L-"H
Medium
High
H-»-L

pcvD
(continued)
(10) 30.0 ± 4.2
(24-38)
(10)
(10)
(10)
(10)
(10)
(10)
(9)
30.4 ± 3.7
(26-37.5)
30.8 ± 2.3
(27.5-34.5)
29.7 r 3.6
(24-34)
30.2 ± 3.1
(26-37)
28.4 ± 3.7
(22-35)
27.6 ± 4.0
(20.5-32)
28.2 ± 1.9
(26-31.5)

- - RBC2)
2.06 ± 0.295)
(1.69-2.33)
2.28 = 0.326)
(1.67-2.67)
2.67 ± 0.16
(2.54-2.91)
2.06 ± 0.31
(1.65-2.49)
2.19 ± 0.58
(1.75-3.54)
1.99 ± 0.32
(1.63-2.61)
1.86 ± 0.367)
(1.31-2.28)
1.91 ± 0.33
(1.48-2.48)

Hb3>
8.2 = 1.25)
(6.8-9.8)
8.9 = 0.96)
(7.6-10.0)
9.8 ± 0.45)
(9.2-10.0)
9.1 ± 1.1
(7.0-10.7)
9.6 ± 1.1
(8.2-11.8)
8.5 ± 1.2
(6.2-10.4)
8.8 ± 1.47)
(7.4-11.2)
9.0 ± 0.8
(8.2-10.4)

 1 Packed cell volume (%)



2)Red blood cells  (106/ul)




3)Hemoglobin (g/dl)



4)
  Liver hemorrhage & necrosis
   =6
                                    218

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Table 66. Red blood  cell  characteristics  in white  leghorn hens  having received
         sludge-fertilized  corn-soybean  diets  from one week of age.

Age and
Relative
Dietarv Cd
50 Weeks
Low


Medium


High


70 Weeks
Low

Medium
High


H-L
L-H
80 Weeks
Low
Medium
High

H-L


L-*H

Mean
Corpuscular
Volume
n"

9
(6)
(3)
9
(6)
(3)
9
(7)
(2)

7

4
7
(2)
(5)
4
10

10
10
5
1
7
1
1
9
1
fl s

152
163
131
139
146
124
146
157
110

135

136
131
156
122
121
127

161
159
159
221
167
206
125
167
95
S.D.

i 19
± 13
± 3
± 13
± 8
± 4
± 26
i 18
± 3

± 14

= 17
± 17
± 0
= 5
± 14
t 10

± 20
± 20
± 10

± 12


i 19

Mean
Corpuscular
Hemoglobin Cone.
n

10


10


9



8

4
7


4
10

10
10
6

9


10

g/dl

28.7


29.3


29.5



32.4

32.9
33.7


32.6
32.9

30.7
29.9
31.6

31.8


32.0

± S.D.

= 1.7


± 1.9


= 1.6



± 1.2

- 5.4
i 1.7


t 1.3
± 1.1

± 1.8
± 2.0
± 2.3

± 1.7


± 1.9

Mean
Corpuscular
Hemoglobin
n

10


10


9



7
1
4
7


4
10

10
10
6

9


9
1
P8 =

39.2


38.4


41.2



41.2
28.7
40.0
39.4


36.7
39.0

45.0
43.1
48.0

47.8


48.0
27.7
S.D.

z 3.8


= 4.1


= 4.9



± 4.9

± 4.0
± 4.6


= 1.9
- 1.7

± 7.5
± 6.6
± 5.3

± 6.5


± 7.5


   Definite outliers  are  listed  separately.   If total group  is  subdivided,  n
   is  parenthetic  (n)  for subgroups.
                                     219

-------
the gizzard may adapt co the Cd insulc.  The consistency of the brain:body
ratio (Table 70) indicated that these differences were not due to body weight
changes.

Liver Parameters—
   The livers of caged layers tend to accumulate large amounts of fat,  espec-
ially when full egg production is approached.   This fatty liver is frequently
friable, inflicted with small hemorrhages and occasionally necrotic in small
areas.  In the 8,  20, and 50 week birds,  histological examination revealed
fatty infiltration or ballooning degeneration.  There was also some degree of
cell swelling.   The histological evaluation generally paralleled gross ob-
servations and gravimetric determinations of % liver fat (Table 71).   Lipo-
fuschin pigment accumulation was assessed in pooled samples of liver at 50,
70, and 80 weeks.   In general,  the changes in liver morphology, fat content,
and pigment accumulation were age related but not influenced by dietary Cd.

   Likewise,  there were age related changes in microsomal protein which were
not influenced by dietary Cd.  Hepatic microsomal cytochrome P-450 was quite
variable,  but apparently depressed by the high dietary Cd (Table 72).   The
variability of these values,  however, is  of some concern and the individual
assays (spectrophotometric tracings)  must be re-evaluated.  There was little
effect of  age or dietary Cd on microsomal NADPH oxidase (Table 72).

   The 0-dealkylation of _p_-nitrophenetole (pNP)  by hepatic microsomes did not
vary significantly with dietary Cd, but there were age changes (Table 73).
The specific activity is listed for 2 pNP concentrations.  Although com-
plete kinetic plots were attempted, these data must also be re-evaluated.
There appeared to be slightly higher affinity (low substrate)  0-dealkylation
activity in the high-Cd birds in spite of decreased amounts of cytochrome P-
450 (Table 72).  The low affinity enzyme  (high substrate concentration) was
not affected.  If the high affinity 0-dealkylase activity is expressed per
nmole P-450,  the differences will probably be significant statistically—
the biochemical significance is unclear.

Conclusions

   1.  Because laying hens consume more feed per unit body weight than other
animals, it is concluded that feed grains produced on soils amended with
sewage sludge,  containing no higher Cd concentrations than those in MSD of
Chicago sludges and applied at recommended N rates, will not affect animal
health and performance.

   2.  At  levels of Cd that are possible  to obtain in the seed or grain of
crops, without serious reductions in yields, there is no evidence that Cd
would interfere with the assimilation of  other nutrients by animals.

   3.  If  levels of Cd in soybeans and corn grain grown on sludge-amended
soils presents a potential hazard to human food-chains,  it is  a nominal one.
Concentrations of Cd in eggs were not affected by levels in corn grain and
soybeans.   Muscle tissues were unaffected by Cd levels in diets.  Considering
offal organs used as human foods,  Cd accumulated at higher concentrations by
order of listing in liver, gizzard, and kidney.   But on a fresh weight basis


                                      220

-------
(including fat and water) Che highest concentration of Cd in liver was about
1.45 ing/kg and well within the upper range reported in animal livers pro-
duced under normal management practices (Kreuzer et al.,  1977).  On a fresh
weight basis, the highest level of Cd in any-tissue was  12.2 rag/kg in kidney.
It therefore appears that the consumption of gizzards and kidneys would pre-
sent about the same health hazard from Cd as diets that  include shellfish.

    4.  The direct consumption of corn grain and soybeans by human animals
must present an exceedingly small potential health hazard.  On a per unit
body weight basis, man does not live long enough to consume as much bio-
logically incorporated Cd as did laying hens.
                                 221

-------
 Table 67.  Leukocyte populations in white leghorn her.s having received sludge-fertilized
           corn-soyoean diets from one week of age.
Age and
Relative
Diecarv Cd
8 Weeks
Low
(range)
Medium
(range)
High
(range)
20 Weeks
Low
Medium
High
50 Weeks
Low
(range)
Medium
(range)
High
(range)
(a)1'
(4)
(3)
(3)
(1)
(8)
(3)
(1)
(8)
(1)
(1)
(9)
(3)
(7)
(1)
(8)
(8)

Segmented
Meutroohils
20.5 = 2.1
(18-23)
14.7 = 4.7
(11-20)
5.7 = 2.1
(4-8)
31
39.9 r 5.2
12.7 r 3.5
68
35.9 = 9.3
20
66
42.7 = 9.4
13.0 z 4.4
37.0 = 11.1
(23-52)
19
33.5 ± 13.3
(13-49)
31.4 : 4.0
(25-37)
Mean = S.D,
Lvmonocvces
72.5 = 3.9
(69-78)
77.7 ± 4.9
(72-81)
88.3 = 3.8
(84-91)
64
53.6 = 9.7
30.7 = 9.0
21
56.6 ± 8.2
73
28
52.2 = 9.9
82.3 = 7.1
52.7 ± 14.0
(27-70)
73
57.4 = 13.9
(38-74)
58.4 ± 7.8
(40-65)
. •%• local Leukocytes
Monocvtes
1.0 = 0.6
(0-2)
0.3 = 0.6
(0-1)
0.3 = 0.6
(0-1)
0
0.4 = 0.7
0
0
0.5 = 1.1
1
0
0.1 = 0.3
0
4.6 ± 3.2
(1-10)
7
4.1 = 3.4
(1-10)
2.9 ± 2.7
(0-7)
Eosinoohils
1.0 = 0
(1-1)
1.3 ± 1.5
(0-3)
0.7 = 1.2
(0-2)
4
1.4 = 1.3
2.0 = 2.0
6
2.8 = 2.2
1
0
0.7 ± 1.4
1.3 = 2.3
1.1 ± 1.1
(0-3)
0
0.6 r 1.1
(0-3)
0.6 = 1.2
(0-3)
Basoohils
5.0 r 2.2
(2-7)
6.0 = 1.4
(5-7)
5.0 = 3.0
(2-8)
1
4.8 r 4.5
4.7 = 5.5
5
4.2 = 5.2.
5
6
4.3 = 3.0
2.3 = 1.5
4.4 ± 3.8
(0-11)
1
4.1 ± 2.7
(0-8)
5.6 ± 6.8
(0-21)

1)
   Treatment groups were occasionally subdivided to emphasize obvious out-liers.  The
   bird at 50 weeks low dose, e.g., was separated due to a large tumor.
                                        222

-------
Table 68.  Serum cneaiscries from whice legnorn hens having received sludge-fertilized
          corn-soybean diets from one week of age.
Age and
Relative
Diecarv
Cd " (n)
Mean
CRT
TP
AL3
p
= S.D.
A?
for Serum"
C?K
G?T LDK
Ca GLU
3 Weeks
Low

Medium

High

(12)

(12)

(11)

1
= 0
1
=0
1
= 0
.3
.2
.5
.2
.3
.3
3
=0
.9
.7
3.6
=0
.5
3.4
= 0.5
7
= 0
7
=0
7
= 1
.5
.9
.4
.9
.4
.2
427
=85
456
= 94
469
=93
146
=58
146
= 36
197
=94
70 431
=18 =108
71 445
±14 =90
59 458
=20 =108
11.3 286
=0.6 =39
11.6 267
=0.7 ±16
11.4 284
=0.5 =22
20 Weeks
Low

Medium

High
(8)

(8)

(8)
1
=0
1
=0,
.3
.5
.6
.7
1.2
±0.4
5.2
±1.1
6.0
= 1.5
4.5
= 1.
4
7
= 1
8
= 1
- 7
±1
.3
.8
.8
.2
.3
.8
>70

>70

>70

185
=117
161
=89
154
= 83
99 >350
±22
122 >350
=53
111 >350
±23
>14 2042>
±125
>14 2192^
= 121
<143) 1722)
±112
50 Weeks
Low

Medium

High

(8)

(8)

(4)









1.0
±0.
4
6.
±1.
5.
±1.
5 .
=1.
3
6
9
1
8
4
— „



8.0
>70

>70
—

_
=2.4
5.9
>70
±2.3
460
= 145
58
±15
66
=16
58
±24
— —

- —

- -

70 Weeks
Low

Medium

High

(7)

(10)

(9)

0.
±0.
0.
=0.
1.
±0.
8
2
9
3
0
4
5 .
±0.
5.
±0.
5 .
=0.
3
8
4
8
2
6
2.33 6.7
=0.29 ±3.
2.57 5.
±0.31 =1.
2.59 6.
=0.45 ±0.
0
9
1
4
9
>70

>70

>70

217
±199
163
=118
118
±54
72
=12
90
±21
88
=22
>153) 225
=11
>153) 230
±10
>153) 244
=13
                                     (continued)
                                     223

-------
 Table  68. (continued)

Age and
Relative
Cd ' (n)
30 Weeks
Low (9)

Medium (9)

High (9)

H-L (10)

L-S (10)

i )
Mean = S.D. for Serum"
CRT

0.7
=0.3
0.7
±0.1
0.7
=0.2
1.0
=0.2
0.8
=0.3
T?

4.9
=0.5
4.6
=0.6
4.7
±0.7
4.8
=0.9
4.6
=0.6
ALB

1.97
=0.28
2.10
=0.32
2.10
=0.32
1.96
±0.24
2.25
±0.22
? A?

6.4 >70
=2.5
7.5 >70
=1.7
6.0 >70
=1.3
6.6 >70
±2.0
5.6 >70
=1.2
C?K

556
=285
555
=196
501
±231
730
=184
599
= 195
G?T

46
= 12
43
=10
51
=14
59
=17
52
=23
LDE Ca GLU

<153) 203
=16
>15 212
±18
<153) 214
= 16
>153) 209
=22
<153) 203
=23

1)
3)
4)
   CRT=Creatinine  (mg/dl);   T?=total  procein (g/dl);  AL3=3lbunen (g/dl); ?=phosphate
   (mg/dl); AP=alkalme  phosphacase;  C?K=creatinine  phospnokinase;  G?T=glutamate-
   pyruvace transaminase; LDH=lactate dehydrogenase  (all  Hycel Incernacional units);
   Ca=calcium  (ng/dl) and GLL"=glucose (ing/dl).
2) .
Low and medium groups each had  one  excessively high glucose value (400) and some
glucose values were quite low at  20 weeks.   Group  means  excluding these values
(deleted values in parentheses) were:

  Low:     282 = 63     (143, 68,  10)             252 ± 21  w/o  400
  Medium:  276 = 67     (52, 40)                  251 = 31  w/o  400
  High:    252 = 22     (56, 44, 14)

Serum Ca was generally  beyond the range of  the method employed  by 20  weeks.
At 20 weeks, the "High  group" had 3 values  between 11-12,  however.  At 70 weeks,
Low and Medium had 1 value each below 13 and High  had 2  low values.   At 80 weeks
Low, L-*H, and High had  2 low values each while H-»-L had one.

n=6
                                      224

-------
Table 69.  Relative liver, gizzard, and kidney weights in voice leghorn hens
          having received sludge-fertilized corn-soybean diets from one week
          of age.

Age and
Relative
Dietarv Cd (n)
3 Weeks
Low
Medium
High
20 Weeks
Low
Medium
High
30 Weeks
Low

Medium
High
50 Weeks
Low
Medium
High
70 Weeks
Low
Medium
High
L-H
H-L
80 Weeks
Low
Medium
High
L+K
H-t-L

(16)
(16)
(16)

(12)
(12)
(12)

(4)

(4)
(4)

(16)
(15)
(16)

(10)
(10)
(10)
(10)
(10)

(10)
(10)
(10)
(10)
(10)



Mean
= S.D". Weight as 7, 3odv
Liver1'


2.10 =
2.18 =
2.39 =


2.07 =
2.25 =
1.96 :


2.40 =


2.34 r
2.

2.
2.
2.

2.
2.
2.
2.
2.

2.
2.
2.
1.
2.
47 =

33 =
74 -
76 =

59 ±
62 ±
57 ±
80 =
76 -

00 =
30 ±
17 ±
98 ±
16 =


0.21
0.16
0.26

0,
0,
0.

0.

0.
0.

0.
0.
0.

0.
0.
0.
0.
0.

0.
0.
0.
0.
0.

.88
.55
.47

,29

,25
,51

59
46
30

64
58
61
74
31

34
74
60
28
49

Gizzard^

2.11 =
2.12 =
2.30 =


1.30 =
1
1

1

1
1

1
1
1

1
1
1
1
1

1
.33 r
.59 =

.13 =

.41 =
.42 =

.16 ±
.27 ±
.33 =

.21 ±
.12 =
.20 =
.12 =
.11 i

.21 ±
1.21 :
1.20 ±
1.14 ±
1.12 r

0.22
0.182)
0.27

0.23
0.162)
0.193)

0.13
3)
0.15J;
0.143>

0.20
0.15
0.193)

0.18
0.16
0.15
0.13
0.16

0.25
0.16
0.12
0.16
0.18

Xidnev1)

0.79 r
0.84 :
0.93 =


0.54 :
0.
0.

0.

0.
0.

0.
0.
0.

0.
0.
0.
0.
0.

0.
0.
0.
0.
0.
,53 =
,54 =

57 :

65 r
62 =

68 =
66 :
69 =

60 ±
64 :
62 ±
58 =
63 =

68 ±
70 ±
68 ±
69 =
66 =

0.08
0.06
0.17

0.06
0.05
0.06

0.04

0.08
0.03

0.13
0.08
0.06

0.06
0.07
0.07
0.06
0.08

0.08
0.12
0.11
0.11
0.08

                                (continued)
                                 225

-------
Table 69. (continued)
   For collective daca of 8, 20, and 30 weeks (n=32) the High Cd group was
   significantly different from the Low Cd and the Medium Cd was
   significantly different fron the High'Cd for ail three tissues at p <0.01.
   A single Low Cd bird nad a very large liver and spleen at 20 weeks (£.68
   and 0.35% body weight, respectively); excluding this bird, the mean
   relative liver weight for n=ll was 1.83 = 0.332 and the mean relative
   spleen weight was 0.15 ± 0.03%.
9)
   Significantly different from High Cd group at p >0.01.

   Significantly different from Low Cd group at p >0.01.
                                 226

-------
 Taole  70.Xeiative organ weigacs in vnice legnorn sens naving received sludge-fertilized
           corr.-soyoean diets from one veetc of age.
Age ana
Relative
Die car/
Cd
3 Weeks
Low
Medium
Sign
20 Veeks
Lou
Medusa
Sign
50 leases
Lov
Medium
Hign
70 Veeks
Low
Medium
Hign
L-a
H-L
30 Weeks
Low
Medium
Hign
L-H
H-L
Mean = S.D. Veignc as Z
(TI)

(16)
(16)
(16)
(12)
(12)
(12)
(16)
(16)
(16)
(10)
(10)
(10)
(10)
(10)
(10)
(10)
(10)
(10)
(10)
Hear-

0.45
O.i5
0.43
0.34
0.34
0.38
0.33
0.42
0.43
0.40
0.36
0.38
0.37
0.39
0.39
0.40
0.39
0.36
0.39


± 0.14
± 0.04
= 0.05
: 0.05
: 0.05
= 0.02
: 0.
: 0.
i 0.
: 0.
: 0.
± 0.
r 0.
= 0.
= 0.
= 0.
: 0.
: 0.
= 0.
07
07
08
07
04
05
08
06
04
03
06
05
09
3rain ?rovencricuii:s

0.41
0.43
0.42
0.2i
0.22
0.23
0.21
0.23
0.22
0.23
0.22
0.22
0.22
0.21
0.22
0.21
0.23
0.22
0.21


± 0.03 0.44 = 0.04
= 0
: 0
: 0
: 0
: 0
: 0
= 0
= 0
: 0
: 0
: 0
: 0
: 0
= 0
: 0
: 0
.03 0.47 : 0.12
.03 0.45 : 0.06
.03 0.26 = 0.04
.02 0.24 : 0.03
.03 0.25 : 0.04
.02
.02
.03
.03
.03
.04
.02
.02
.03
.03
.02
± 0.03
: 0.03
3odv

Pancreas

0.23
0.34
0.34
0.13
0.18
0.19
0.20
0.24
0.25
o.:o
0.22
0.22
0.22
0.20
0.19
0.20
0.23
0.21
0.19

± 0.03
= 0.04
: 0.04
: 0.03
: 0.03
= 0.03
= 0.03
= 0.02
= 0.04
: 0.02
= 0.03
: 0.03
= 0.05
: 0.02
: 0.05
: 0.03
: 0.02
± 0.02
= 0.04


Soiaen

0.20
0.21
0.22
0.17
0.13
0.16
0.081
0.082
0.085
0.087
0.083
0.081
0.082
0.075
0.111
0.080
0.079
0.084
0.091

= 0.04
= 0.04
: 0.06
: 0.071}
: 0.02
: 0.05
: 0.014
; 0.019
: 0.019
: 0.032
: 0.031
= 0.014
= 0.016
: 0.015
± 0.051
r 0.024
= 0.018
± 0.014
= 0.024
1)
   For collective daca of 3, 20, and 30 weeks (n-32) che Hign Cd group was  significantly
   different from che Low Cd and che Medium Cd was significantly different  from  che  Hign
   Cd for all three tissues ac p <0.01.  A single Low Cd bird had a very  large liver and
   spleen ac 20 veeks (4.63 and 0.35Z body weighc, resoectively); excluding this  bird,
   che mean relative liver weignc for n-11 was 1.33 = 0.33Z and c.ne mean  relative spleen
   weignc was 0.15 : 0.03Z.
                                     227

-------
Table 71.  Fat concent of livers from white leghorn hens having received
          sludge-ferciiized corn-soybean diecs from one weex of age.
Age and Relative
Die car-/ Cd n
8 Weeks
Low
Medium
High
20 Weeks
Low
Medium
High
30 Weeks
Low
Medium
High
50 Weeks
Low
Medium
High
70 Weeks
Low
Medium
High
H-L
L-H
80 Weeks
Low
Medium
High
H-"L
L-H
15
13
15
12
12
12
4
4
2
16
16
16
10
10
10
10
10
10
9
10
10
10
Liver
: S
fresh
4.81
4.38
4.46
12.3
21.6
13.9
10.0
9.3
13.4
9.9
12.0
12.8
13.9
18.1
15.3
15.8
15.9
9.6
11.0
12.5
8.1
7.8
Fac (aean
.D. Z
weignt)
= 0.75
: 0.61
: 0.96
± 6.0
= 9.8
= 8.4
± 3.7
- 4.6
r 4.9
± 4.1
= 4.0
: 5.9
± 6.5
= 11.1
r 6.6
± 5.9
± 7.3
± 6.9
: 6.8
± 8.6
= 4.8
r 4.2
                                  228

-------
Table 72.  Hepatic nicrosoraal parameters in white leghorn hens having received
          different levels of dietary Cd from one ween of age.

Necr
Time
8 wk


20 wk


30 wk


50 wk


70 wk




80 wk


Dose
GrouD
Low
Med
High
Low
Med
High
Low
Med
High
Low
Med
High
Low
Low-High
Med
High-*Low
High
Low
Med
High
Total
28
24
22
10
9
10
.2
.5
.0
.65
.43
.56
12.55
12.43
11.97
15.
17.
15.
17.
14.
17.
15.
15.
15.
16.
16.
,86
.13
,99
73
39
04
62
42
60
51
85
Protein
Liver
= 6
= 3
.9
m 2
= 5.9
= 1.2
± 1.
= 2.
: 2.
= 2.
± 2.
- 2.
± 1.
= 2.
- 2 .
= 2.
= 2.
± 2.
= 2.
= 2.
± 1.
= 2.
.3
.2
,6
,4
0
7
4
2
5
9
6
7
4
9
86
4
P-450
^tn/mer Protein
.280 =
.241 z
.176 r
.159 =
.184 =
.136 =
.112 =
.119 =
.122 =
.243 ±
.181 =
.219 =
.233 =
.252 =
.186 r
.209 =
.191 ±
.200 ±
.140 =
.135 =
.084
.119
.057
.098
.059
.018
.075
.051
.039
.019
.089
.079
.16
.17
.08
.13
.12
.09
.056
.071
NADPH
jn/niin/me Protein
.0316=
.026 =
.030 =
.025 =
.027 =
.028 =
.026 =
.027 =
.029 =
.023 =
.020 =
.024 =
.030 =
.039 =
.034 i
.037 =
.038 t
.028 ±
.022 ±
.029 ±
.006
.006
.008
.010
.008
.004
.001
.003
.008
.007
.009
.006
.008
.005
.005
.009
.006
.010
.008
.006
                                 229

-------
Taole 73.  Hepacic aicrosonal 0-dealkyiacion of £-nicrophenecois ir. whice
          leghorn hens having received dietary Cd from one week of age.

Age ana Relative
Diecarv Cd n
3 Weeks
Low
Medium
Hign
20 Weeks
Low
Medium
High
50 Weeks
Low
Medium
High
70 Weeks
Low
Medium
High
80 Weeks
Low
Medium
High
5
4
4
5
5
8
7
9
8
5
6
6
6
6
6


1
1
1
1
1
1
0,
0,
1.
1.
1.
1.
1.
1.
1.
n
0.
.55
.52
.70
.57
.42
.61
.85
.90
.06
,09
,16
18
22
02
18
moles/ainuce/mE orocein
05 aM- -
= 0
= 0
= 0
= 0,
= 0.
= 0.
= 0.
= 0.
= 0.
= 0.
± 0.
= 0.
i 0.
= 0.
= 0.
.43
.37
.21
.55
.63
.30
.19
20
26
26
25
12
33
25
32
2.
0 aM
4.45 = 1
4
4
3
2
3
2
1
2

2,
2,
2.
2.
2.
.08
.75
.32
.90
.15
.10
.96
.35

.61
.71
,56
,21
61
= 1
= 0
= 0
= 0
= -
= 0
± 0
: 0,

= 0.
= 0.
r 0.
: 0.
= 1.

.03
.08
.38
.67
.26
.06
. 55
.52
.46

,32
,40
58
33
00
                                  230

-------
                                     SECTION  6

                           ABSTRACT  OF  THESIS  RESEARCH
 CADMIUM-INDUCED GROWTH DEPRESSION AND  CADMIUM ACCUMULATION IN CHICKS AS
 INFLUENCED BY DIETARY MODIFICATIONS

   Nine experiments were conducted with week-old crossbred meat-type chicks
 co explore the effect of dietary nutrient balance upon the response of chicks
 to cadmium.  When 1, 5, 10, 15, 20, 30, and &0 ppm of Cd as Cd/Cl, was added
 to corn-soya diets for two weeks, there was growth depression at 10 ppm and
 a dose-related increase in the Cd content of kidneys and livers due to added
 Cd.  As the Cd intake was increased, there was an increase in the percent of
 ingested Cd retained in these two tissues.

   Simultaneous decreases in dietary Ca, Zn, ?, and Mn increased the growth
 sensitivity of chicks to added Cd and  increased Cd retention without affect-
 ing growth when no Cd was fed.  When the diet was adjusted to be marginal in
 methionine and Mn, supplementing the diet with either of these nutrients did
 not influence growth or Cd retention when Cd was added to the diet.  Added Cu
 did not alleviate Cd-induced growth depression, but increased Cd retention.
 Supplemental Zn and levels of Ca above accepted requirements ameliorated Cd-
 induced growth depression and reduced  the amount of Cd retained in livers and
 kidneys.

   Chicks depleted of vitamin D during the first week of life were found to
be more growth-sensitive to 10 ppm of Cd in subsequent 2-week assays.   High
levels of vitamin D reduced the growth depression and increased liver and
kidney accumulation of Cd in both depleted and undepleted chicks.   When
levels of Ca and virgin D that were marginal for maintaining normal bone
ash were fed,  10 ppm of added Cd reduced tibia bone ash.
                                 231

-------
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