Federal Water Pollution Control Administration
         Division of Water Quality Research
        Analytical Quality Control Laboratory
                Cincinnati, Ohio
USE OF A  FLOATING  PERIPHYTON  SAMPLER

   FOR WATER  POLLUTION SURVEILLANCE

                    Reprint
                 February 1970
         U.S. DEPARTMENT OF THE INTERIOR

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USE OF A FLOATING PERIPHYTON SAMPLES
  FOR WATER POLLUTION SURVEILLANCE
      Cornelius I. Weber, Ph.D.
                 and
          Ronald Lo Raschke
               Reprint
            February 1970

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                            Preface


     This report was first published on September, 1966 as

Applications and Development Report No. 20 of the Water
                ^
Pollution Surveillance System.  The laboratory that operated

the Water Pollution Surveillance System was subsequently

transferred to the Office of Research and Development, and

was renamed the Analytical Quality Control Laboratory.

     At the time this report was written, Ronald Raschke was

an aquatic biologist in the plankton laboratory.  He later

received his doctorate from Iowa State University, Ames, and

is now on the staff of the Botany Department, Rutgers University,

New Brunswick, New Jersey.
                          Cornelius I. Weber, Ph.D.
                          Chief, Biological Methods
                          Analytical Quality Control
                                  Laboratory

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                         Table of Contents



                                                                Page



1.  Introduction 	    2



2.  Methods and Equipment	    3



3.  Bate of Colonization	    5



k.  Comparison of Plankton and Periphyton  	    9



5.  Dry and Ash-Free Weights of Periphyton ..........   13



6.  Periphyton Diatoms as Pollution Indicators 	   16



7.  Literature Cited 	   20

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                              Figures



                                                             Page



1.  The periphyton sampler •	   4



2.  Density of live diatoms in periphyton and plankton



    samples from the Ohio River at Cincinnati, October



    20 - November 21, 19&-	   6



3*  Rate of colonization of glass slides by pennate diatoms



    in the Ohio River at Cincinnati, October 20 - November



    21, 19&	   7



k.  Rate of colonization of glass slides by centric diatoms



    in the Ohio River at Cincinnati, October 20 - November



    21, 19&	   8



5«  Dry and ash-free weights of scrapings from glass slides



    exposed 2 weeks in the Ohio River at Cincinnati, July 3



    to December 10, 1965	I1*-

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                             Tables



                                                            Page



1*  Dominant diatom species in perlphyton and plankton



    samples from the Ohio River at Cincinnati, October



    20 - November 21, 19&	   10



2.  Live Centric to Live Pennate cell ratios in periphyton



    and plankton samples from the Ohio River at Cincinnati   11



3*  Percent dead diatom cells in periphyton and plankton



    samples from the Ohio River at Cincinnati  	   12



k.  Dry and ash-free weights of periphyton samples from



    the Ohio River at Cincinnati	   15



5.  The most abundant diatom species in periphyton samples



    collected in August 1965 above and below a pollution



    source in the KLamath River, Oregon  .........   17

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               Use of a Floating Periphyton Sampler

                 for Water Pollution Surveillance
                             Abstract





     A floating sampler was used to collect periphyton in the Ohio



River at Cincinnati.  The rate of colonization of glass slides by



diatoms was determined, and the periphyton and plankton diatom



communities were compared.  The density of live diatom cells on the


                            2
slides reached 15,000 per mm  in approximately 30 days.  The peri-



phyton diatoms were dominated by species of Nitzschia and Navicula,



whereas the plankton diatoms consisted largely of species of



Melosira and Cyclotella.  Dry weights of scrapings from slides

                                                       n

exposed 1^4- days ranged from 1^9 • 5 mg per slide (32<>5 cm ) in July



1965, to 2.7 mg per slide in December 1965.  Ash-free weights



averaged 16.2$ of the dry weight.



     Glass slides were exposed in the floating sampler above and



below a group of polluted outfalls on the upper KLamath River,



Oregon.  Gomphonema parvulum and Nitzschia palea were the most



abundant diatoms in samples taken in the vicinity of the pollution



sources; whereas Cocconeis placentula was dominant above the out-



falls, and in the oligosaprobic zone downstream.

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                                -2-



                           Introductiou



     Exploratory periphyton studies were Initiated "by the Water



Pollution Surveillance System of the U. S. Public Health Service



in the Fall of 19& to augment phytoplankton data collected from



a nation-vide system of approximately 135 stations*



     Man;r aquatic "biologists (FJerdinstad, 196k; Hynes, 19^3;



Novak, 19l*0) have recognized that a satisfactory interpretation



of phytoplankton data obtained for pollution studies from videly



separated river stations is rarely achieved.  Attempts to relate



the quality and quantity of algae in grab vater samples to known



or suspected types of pollution are usually confounded by an igno-



rance of the origin and physiological condition of the organisms.



In contrast, the presence of significant quantities of attached



algae on natural or artificial substrates is strong evidence of



the suitability of the vater for the growth of the organisms col-



lected at a station.  Inferences regarding vater quality can be



formulated vith greater confidence therefore, if they are based on



the composition and density of the periphyton.  Periphyton communities



have long been used by European biologists to characterize pollution



(Kolkvitz and Marsson, 1908; Butcher, 19«*-6, 19^7, 19^9, 1955; FJer-



dinstad, 1950, 1964; siadeckova, 1962; Sladeckova and Sladecek, 1963).



Interest in the periphyton developed later in this country (Blum,



1957; Hohn, 1959; Patrick, 1953, 1957).  Except for the vork of Neel



(1953)> the periphyton has received little attention In Federal



pollution studies in the United States.

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                                -3-

                       Methods and Equipment

     The sampler consists of a styrofoam float approximately

12 X 12 X 2 in., supporting a central plexiglass cradle holding

1- X 3-in. glass microscope slides (Figure 1).  The construction

materials are commercially available and cost approximately $2.50.

The slides are held with their long axes parallel to, and their

short axes perpendicular to, the water surface.

     Slides were removed from the sampler after exposures of 1, k,

7, 15, and 32 days.  The periphyton was scraped from the slides

with a razor blade, arid counts were made to determine number of
            p
cells per mm  of slide area (Weber, 1966).  Live and dead diatom

cells were identified to genus on millipore filter preparations

examined at 1000X.  Species determinations were made from permanent

(Hyrax) mounts of incinerated diatom materials.

     Samples used to determine dry and ash-free weights consisted of

material from four replicate slides, each treated as a separate sub-

sample.  These slides were allowed to air dry in the field.  The

material was further dried at 50 C in the laboratory and stored in

sealed containers until used.  When processed, the slides were

wetted with distilled water, scraped, and the scrapings dried 2k

hours at 105°C and fired for 1 hour at 500°C.

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Figure 1.  The periphyton sampler.

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                                -5-

                       Rate of Colonization

     Following a brief initial lag period, the density of live diatom

cells on the slides increased exponentially, reaching approximately
             p
15,000 per mm  in 32 days  (Figure 2).  This rapid rate of increase

indicated that the colonization resulted primarily from the division

of cells which had became attached to the slides during the first few

days of exposure (during the lag period).  Diatom counts in the

plankton taken near the sampler remained relatively constant during

the study period.  Had the colonization of the slides resulted prin-

cipally from the gradual deposition of drifting cells, a linear rise

in density would have been observed.

     Judging from the decline in the growth rate during the latter

part of the exposure period, it was assumed that the population was

largely established within 15 days, and had leveled off at approx-
                     P
imately 15,000 per mm  (32 days).  This is similar to the cell density

that Butcher (19^6) found on glass slides exposed 30 days in eutrophic

(oligosaprobic) waters.

     It was decided, on the basis of the cell density data, to tenta-

tively adopt a two-week exposure period for the collection of periphyton

samples at Water Pollution Surveillance System stations.  This period

would be long enough to permit the development of a populous periphyton

community, yet brief enough to reflect short-term changes in the

water quality.

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                                 -6-
                                            D
                             PERIPHYTON

                             O  TOTAL DIATOMS
                                 PENMATES
                                 CENTRICS
                             PLANKTON
                              D TOTAL DIATOMS
                                                    _l
                                                    z

                                                 1C? CO
                                                 K?
                                                    LU
                                                    O
                         O
                                                 id  CL
                       10
 15   20   25   30  35

DAYS
Figure 2.  Density of live diatoms in periphyton and plankton samples
from the Ohio River at Cincinnati, October 20 - November 21,

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                           -7-
       4
      10
OJ.
  CO
  _J
  _)
  LU
  O
      10
      ,0°
                                 O
                                 A
                                 D
NITZSCHIA
NAVICULA
ACHNANTHES
GOMPHONEMA
SYNEDRA
                   10    15   20   25   30   35
                          DAYS
        Figure 3.  Rate of colonization of glass slides
        by pennate diatoms in the Ohio River at-Cincinnati,
        October 20 - November 21,

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                           -8-
CVJ
  —^


  CO
  _J
  bJ
  O    i
      10
      ,0°
                                   MELOSIRA
                                   CYCLOTELLA
              5    10    15    20   25   30   35
                          DAYS
          Figure k.  Rate of colonization of glass slides
          by centric diatoms in the Ohio River at Cincinnati,
          October 20 - November 21,

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                                 -9-
     The periphyton diatom population was dominated by pennates
during the entire period,  They  comprised 67$ of the diatoms at 7
days, and approximately 85$ at 15 and 32 days.  Gomphonema and
Synedra were the most abundant pennates at 7 days, but the density
of both was later exceeded by Nitzschia, Navicula, and Achnanthes
(Figure 3)«  The dominant species were Nitzschia paradoxa Gmel.,
Navicula cryptocephala Kutz., N. tripunctata (0. Mull.) Bory,
Gomphonema parvulum (Kutz.) Grun., G. olivaceum (Lyng.) Kutz.,
Synedra ulna (Nitzsch) Ehr., and Achnanthes sp.  The abundance curves
of the centric diatoms are shown in Figure 4.  The principal species
were Melosira varians Ag., and Cyclotella meneghiniana Kutz.

              Comparison of Plankton and Periphyton
     Although colonization of the slides was unquestionably pioneered
by "drifting" organisms, the composition of the periphyton and plankton
diatom communities was very different throughout the entire exposure
period (Table l).  In contrast to the periphyton, the plankton diatoms
were principally Gentries, consisting of Melosira ambigua (Grun.) 0.
Mull., M. distans (Ehr.) Kutz., and species of Stephanodlscus and
Cyclotella.  This was further evidenced by the ratios of live centric
and pennate cells in the two types of samples shown in Table 2.
     The proportions of live and dead diatom cells in the two types
of samples were also of interest (Table 3).  Since the organic matter
must be removed from the diatom frustules before species Identifica-
tions can be made, it is not possible to distinguish which cells in

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Table 1.  Dominant diatom species in perlphyton and plankton samples from the Ohio River at

1 Day
4 Days
Exposure Period
7 Days
15 Days
32 Days
Periphyton
Synedra
ulna.
Meloslra
granulata
Nitzschla sp.
Navicula sp.
Nitzschla
paradoxa
Melosira
varians
Synedra
ulna.
olivaceum
Gomphonema
parvulum
Synedra
ulna.
Melosira
varians
Nitzschla sp.
Gomphonema
parvulum
Melosira
varians
Navicula
cryptocephala
Nitzschla
paradoxa
Meloslra
varians
Nitzschla
paradoxa
Navicula
tripunctata
Navicula
cryptocephala
Plankton
Meloslra
ambigua
Cyclotella
meneghlnlana
Unknown centric
invisitatus
Stephanodiscus
hantzschli
Meloslra
ambigua
Cyclotella
meneghlnlana
Unknown centric
Melosira
ambigua
Melosira
dlstans
Fragilarla
crotonensis
Cyclotella
meneghlnlana
Melosira
dlstans
Melosira
ambigua
Meloslra
granulata
Unknown centric
Melosira
dlstans
Asterionella
formosa
Melosira
ambigua
Unknown centric

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                                    -11-
Table  2,   Live Centric to Live Pennate cell ratios in periphyton and plankton
                 samples from the Ohio River at Cincinnati
Periphyton
Period Exposed
10/20-21/64 (1 day)
10/20-24/64 (4 days)
10/20-27/64 (7 days)
10/20-11/4/64 (15 days)
10/20-11/21/64 (32 days)
Live Live
r!ATvhi*1 f* •T^pnnfl'f^A

1:2.4
1:4.2
1:2.0
1:6.4
1:10.7
Plankton
Date
10/20/64
10/24/64
10/27/64
11/4/64
n/2i/64
Live Live

3.7:1
5.2:1
28.5:1
2.0:1
0.7:1

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                              -12-
Table  3.  Percent dead diatom cells in periphyton and plankton
            samples from the Ohio River at Cincinnati
Periphyton
Period Exposed
10/20-21/64 (1 day)
10/20-24/64 (4 days)
10/20-27/64 (7 days)
10/20-11/4/64 (15 days)
10/20-11/21/64 (32 days)
% Dead
9
18
2
23
9
mean 12
Plankton
Date
10/20/64
10/24/64
10/27/64
11/4/64
11/21/64
$ Dead
34
35
48
30
12
nean 32

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                               -13-

the permanent mounts vere alive or dead vben the samples were taken.

We have observed that plankton samples usually contain a high per-

centage of dead diatom cells.  The mean proportion of dead diatom

cells In plankton samples taken twice Monthly at this station during

the k water years October 1, I960 to September 30, 19& was 29.6)6,

and the proportion of dead diatom cells in the plankton samples

collected during this study averaged 32$.  This is a serious weak-

ness in the data, for although it is not likely that the majority

of the cells of the dominant species in a sample would be dead, the

possibility cannot be discounted.  Therefore, a high degree of un-

certainty is associated with any interpretation of plankton diatom

species data.

     The percentage of dead diatom cells in the periphyton was much

lower, however, averaging only 12$ during the 32-day period dis-

cussed above (Table 3), and 15.6)6 in 12 two-week samples taken in

during the 1965 calendar year.  The high   percentage of live diatom

cells in the periphyton confers a significantly greater reliability

upon the inferences based on the diatom data from these samples.



              Dry and Ash-free Weights of Periphyton
                                                        n
     Dry weights of scrapings from slides (area, 32.5 cm ) exposed

for two-week intervals from July 3 to December 10, 19&5, decreased

from 1^9.5 mg per slide to 2.8 mg per slide, and ash-free weights

decreased from 21,8 mg per slide to 0.3 mg per slide (Figure 5).

The proportion of organic matter in the samples, however, remained

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8
X

u.
<





UJ
0
rt
\



30
20
5
0 10

0
200
180
160

140
120

100
80
60
40
20
•
" "
• • • • -
•
.


* ^
•
.
i
»
.



j


< o DW
• AFW
•^

1
P
I
•1 i- ' 1
0
                                             N
Figure 5.  Dry and ash-free weights of scrapings from glass
        slides exposed 2 weeks in the Ohio River at
         Cincinnati, July 3 to December 10, 1965.

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                                 -15-
Table 4 .   Dry and ash-free weights of periphyton samples from the Ohio
                         River at Cincinnati
Period
Exposed
7/3-7/17/65
7/17-7/31/65
8/14-8/30/65
8/30-9/14/65
10/30-11/14/65
11/26-12/10/65
Dry Wt
(mg),
149.5^3.^
145.6*33-1
73.8tl4.8
48.2120.8
16.5-9.6
2.lto.7
C.V.*
(*)
37.5
22.7
20.0
43.2
56.4
33-3
Ash-Free Wt
(mg)
21.8*6.4
16. 7^3. 6
9.5tl.8
7.ll3.6
4.8t2.7
0. 3^0.2
c.v.
(*)
29.4
21.3
18.9
50.7
56.2
66.7
Ash-Free Wt
Dry Wt
.146
.115
.129
.147
.291
.143
                                                Mean
                                                            .162
      coefficient of variation.

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                               -16-




relatively constant, averaging 16.2$ (Table *»•).  This was consider-




ably lower than the proportion of organic matter found in periphyton




by Newcombe (19^9, 1950) and Nielson (1953), and in the seston of



Wisconsin lakes by Birge and Juday (1922, 193^).  The proportion of




organic matter in dried algal cells usually ranges between 50$ and



90$.  The value obtained for the periphyton in our study was similar



to that reported by Nelson and Scott (1962) for the seston (12.9$)




in the Oconee River.  We have no data on the organic content of the



seston in the Ohio River at Cincinnati.  However, the organic con-




tent of the seston in ^9 weekly samples from the nearby Little



Miami River, taken during the 196^-65 water year, averaged 15.8$.



It is likely, therefore, that the major portion of the organic



matter which accumulated on the slides was derived from the seston



(evan though the periphyton was shown to be very different from the



plankton).  It was concluded that dry and ash-free weights did not




accurately measure the production of organic matter by periphyton



growing on slides at this station.






            Periphyton Diatoms as Pollution Indicators



     The utility of periphyton in characterizing pollution is



illustrated by the data obtained from three stations on the upper



reaches of the KLamath River near Klamath Falls, Oregon.  These



samples were supplied by personnel from the Klamath Basin Project,



as a part of a cooperative study.

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                                    -17-
Table 5.  The most abundant diatom species in periphyton samples collected
            in August 1965 above and below a pollution source
                       in the KLamath River, Oregon
Station .1
(7/23-8/6/65)
Species
Cocconeis
placentula
Navicula
Cryptocephala
Nitzschia
oregona
Gomphoneis
herculeana
Others
%
Abun-
dance
28
22
13
9
28
Cells
Per
	
160
125
Ik
51
502
Station 2
(7/23-8/6/65)
Species
Gomphonema
parvulum
Nitzschia
(palea)
Cocconeis
placentula
Nitzschia
oregona
Others
%
Abun-
dance
44
21
18
5
12
Cells
Per
nm2
2113
1008
864
240
4686
Station 4
(7/9-8/6/65)
Species
Cocconeis
placentula
Nitzschia
oregona
Stephanodis cus
invisitatus
Nitzschia
amphibia
Others
%
Abun-
dance
39
9
6
6
40
Cells
Per
mm,
*
-
-
-
-
*A quantitative comparison of cell densities with this station was
 not possible because of the difference in length of the exposure
 period.

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                               -18-



     The general pattern of water quality at the stations can be



established on the basis of the diatom populations alone, without



other knowledge of environmental conditions (Table 5)»  The domi-



nance of Cocconeis placentula (Ehr.) at Station 1, above KLamath



Falls, indicates an abundance of inorganic nutrients and a low



(or moderate) level of dissolved organics.  At Station 2, just



below the city, the dominance of Gomphonema parvulum (Kutz.) and



Nitzschia palea (Kutz.) W. Sm. indicates high levels of dissolved



organics (gross organic pollution).  The reoccurrence of Cocconeis



placentula as the dominant form at Station ^,30 miles below the



city, is indicative of a return to nearly oligosaprobic conditions,



resulting from oxidation of the organics.



     Station 1 is located at the mouth of the Link River, which



drains Upper Klamath Lake, a eutrophic lake with a long history of



nuisance algal blooms.  The abundance of Cocconeis placentula at



this station is in agreement with the distribution pattern of this



organism found by Butcher (19^7)> who reported it as a dominant



diatom in oligosaprobic (and eutrophic) waters.  Fjerdinstad (1950)



found it in alpha- and beta-mesosaprobic habitats also, which would



explain its occurrence at Station 2, located less than two mi.les



below sewage treatment plant, tallow works, and wood processing



industry waste outfalls.  The dominance of Gomphonema parvulum and



Nitzschia palea at Station 2 is a direct result of the effects of

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                              -19-



the organic pollution.  Butcher (19^7) found these two diatoms to



be the most resistant to pollution, and Fjerdinstad (196^) refers



to them as saprophilous, "Occurring most generally in polluted



waters...".



     Irrigation return water from the Lost River Basin is discharged



into the Klamath River approximately 11 miles below Station 2.



Another 7 miles downstream the river enters a narrow gorge and



tumbles approximately 2 miles over a rocky bed, falling 300 feet.



Station 4 is located 2 miles below this rapid.  Here, Cocconeis



placentula was again the most abundant diatom, accounting for 30$



of the diatom population, whereas Nitzschia palea and Gomphonema



parvulum each comprised only 3$ (included under "others" in Table 5)»



The natural aeration caused by the rapids undoubtedly aided oxidation



of dissolved organics and restoration of oligosaprobic conditions.



     The usefulness of the periphyton in determining water quality



is supported by an extensive literature concerning the ecology of



the organisms, which has accumulated during many decades of work by



European aquatic biologists.  The examples cited above employed only



the dominant diatom species.  A more precise determination of con-



ditions at these stations could have been made by describing the



entire periphyton community.

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                               -20-



                         Literature Cited



Birge, E. A., and C. Juday.  1922.  The inland lakes of Wisconsin.



  The Plankton.  I. Its quantity and chemical composition.  Wis.



  Geol. Nat. ELst. Surv., Bull. No. 64, Sci. Ser. No. 13. 219 PP»






	.  1934•  Partlculate and dissolved organic matter in inland
  lakes.  Ecol. Monogr. 4:






Blum, J. L.  1956.  The ecology of river algae.  Bot. Rev. 22(5):



  291-341.





	.  1957.  An ecological study of the Saline River, Michigan.
  flydrobiologia 9:361-408.





Butcher, R. W.  1946.  Studies in the ecology of rivers.  VI. The



  algal growth in certain highly calcareous streams.  J. Ecol.



  33:268-283.





	.  1947.  Studies in the ecology of rivers.  VII. The algae of



  organically enriched waters.  J. Ecol. 35:186-191.





	.  1949.  Problems of distribution of sessile algae in running
  water.  Verh. Int. Ver. Limnol. 10:98-103.
      .  1955*  Relation between the biology and the polluted condition
  of the Trent.  Verh. Int. Ver. Limnol. 12:823-827.





FJerdinstad, E.  1950.  The microflora of the River MQlleaa, with



  special reference to the relation of the benthal algae to pollution.




  Folia Limnol. Scand. 5:1-123.

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                               -21-



FJerdinstad, E.  19&.  Pollution of streams estimated by benthal



  phytomicro-organisms.  I. A saprobic system based on communities



  of organisms and ecological factors.  Int. Rev. ges. ffydrobiol.




  ^9:63-131.





Hohn, M. H.  1959*  The use of diatom population as a measure of



  water quality in selected areas of Galveston and Chocolate Bay,



  Texas.  Xnst. Mar. Sci. 6:206-212.





Bynes, H. B. N.  1963.  The biology of polluted waters.  Liverpool,



  Liverpool Univ. Press, 202 pp.





Kolkwltz, R., and M. Marsson.  1908.  Okologie der pflanzlichen



  Saproblen. Ber. deut. Bot. Ges. 26:505-519.





Neel, J. K.  1953.  Certain limnological features of a polluted



  irrigation stream.  Trans. Amer. Microsco. Soc. 72:119-135.





Nelson, D. J., and D. C. Scott.  1962.  Role of detritus in the



  productivity of a rock-outcrop community in a piedmont stream.



  Limnol. Oceanogr. 7:396-^13.





Newcombe, C. L.  19^9.  Attachment materials in relation to water



  productivity.  Trans. Amer. Microsc. Soc. 68:355-361.





	.  1950.  A Quantitative study of attachment materials in
  Sodon Lake, Michigan.  Ecology 31:20*1-215.

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                               -22-



Nielson, R. S.  1953*  Apparatus and methods for the collection of



  attachment materials in lakes.  Progr. Fish-Cult. 15:87-89.






Novak, W.  19^0.  Uber die Verunrelnigung eines kleinen Flusses in



  Mahren durch Abwasser von Weissgerbereien, Leder- und Leimfabriken



  und anderen Betrieben.  Arch. Hydrobiol. 36:386-423.





Patrick, R.  1953*  Aquatic organisms as an aid in solving waste



  disposal problems.  Sew. Ind. Wastes 25:210-217.






	.  1957*  Diatoms as indicators of changes in environmental
  conditions.  Syrap. Biological Problems in Water Pollution.  U6PH5,



  Cincinnati, pp. 71-83.





Sladeckova, A.  1962.  Limnological investigation methods for the



  periphyton ("Aufwuchs") community.  Bot. Rev. 28:286-350.





	, and V. Sladecek.  1963*  Periphyton as indicator of the



  reservoir water quality.  I. True-periphyton.  Tech. Water



  7:507-561.





Weber, C. I.  1966.  Methods of Collection and Analysis of Plankton



  and Periphyton Samples in the Water Pollution Surveillance System.



  Water Pollution Surveillance System Applications and Development



  Report No. 19, 19 pp.

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