SEPA
                                  United States
                                  Environmental Protection
                                  Agency
                                  Industrial Environmental Research
                                  Laboratory
                                  Cincinnati OH 45268
                                  Research and Development
                                  EPA-600/S2-82-060   Oct. 1982
Project  Summary
                                 Evaluation  of  Waste  Citrus
                                 Activated  Sludge in
                                 Poultry  Feeds
                                  Bobby L. Damron, Douglas M. Janky, Robert H. Harms, and
                                  Mark F. Hall
                                   Experiments  were  conducted on
                                  chick broilers and hens to determine
                                  the metabolizable energy of citrus
                                  sludge. A determination of metaboliz-
                                  able energy values showed that the
                                  values decreased as the level of citrus
                                  sludge in the diet increased. A series of
                                  protein levels were fed to day-old
                                  chicks for a three-week period in an
                                  attempt to assay the protein quality of
                                  citrus sludge. As the level of sludge in the
                                  diet increased, its utilization decreased,
                                  with  an almost toxic effect being
                                  observed at higher concentrations.
                                   Vitamin Da and sludge were fed to
                                  day-old chicks  to determine if the
                                  minerals contained in the sludge could
                                  be causing the destruction of Vitamin
                                  D3 in the feed.  Results revealed that
                                  the addition of Vitamin  D3 did  not
                                  offset the depressing effect of sludge
                                  feeding upon  body  weights. The
                                  conclusion reached was that citrus
                                  sludge was not  related to Vitamin D3
                                  destruction.
                                   Experiments  were  conducted on
                                  day-old chicks to evaluate the biological
                                  availability of the phosphorus contained
                                  in citrus sludge.  All sludge treatments
                                  produced body  weights significantly
                                  below those of a modified corn-soy
                                  basal diet containing monosodium
                                  phosphate.
                                   Citrus sludge was also examined as
                                  a source of pigmentation in egg yolks.
                                  Hens  were fed  a white corn basal,
                                  xanthophyll-free diet to deplete egg
                                  yolk pigmentation. It was observed
                                  that citrus sludge produced eggs with
                                  the same hue as yellow corn containing
                                  comparable amounts of xanthophyll.
                                  The xanthophylls  of yellow corn
                                  appeared to  be more biologically
                                  available than those in citrus sludge.
                                    Broilers were used to determine the
                                  effectiveness  of citrus sludge as a
                                  pigmenting agent. White corn diets
                                  mixed with sludge were compared with
                                  yellow  corn diets. Feeding  sludge
                                  resulted in both the shanks and skin
                                  imparting the same hue and darkness
                                  as feeding yellow corn at equivalent
                                  xanthophyll levels.
                                    This Project Summary was de vel oped
                                  by EPA's Industrial Environmental
                                  Research Laboratory, Cincinnati, OH,
                                  to announce key findings  of the
                                  research project that is fully docu-
                                  mented in a separate report of the
                                  same title (see Project Report ordering
                                  information at back).

                                  Introduction
                                   Waste citrus activated sludge is the
                                  product resulting from the treatment of
                                  citrus processing plant wash water in a
                                  facility similar to a municipal treatment
                                  plant. One  of the major problems
                                  associated with this process  is the
                                  production of excess activated sludge.
                                  The  handling and disposal of  sludge
                                  amounts to one of the larger operating
                                  costs of the system. A typical analysis of
                                  the sludge is presented in Tables 1 and
                                  2.
                                   From previous studies with  chicks
                                  (broilers) and hens (Jones et al., 1975),
                                  the recovery of activated sludge for the

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 Table 1.    Proximate Analysis of Citrus
           Sludge
Nutrient
Moisture
Crude Protein
Fat
Fiber
Ash
NFE
Calcium
Phosphorus
Xanthophyll
Carotene
Percent
11.10
22.35
7.40
11.45
12.20
35.50
1.60
0.45
28. 60 mg/ kg
11.00mg/kg
Table 2.    Trace Mineral Analysis of
           Citrus Sludge
Element
Aluminum
Cadmium
Chromium
Cobalt
Copper
Iron
Lead
Magnesium
Manganese
Mercury
Nickel
Potassium
Selenium
Sodium
Zinc
Content
(ppm)*
2,500
1
100
3
260
4,600
120
2,100
51
0.5
120
7,300
<1
5,300
430
* Dry weight basis (ppm - mg/kg).

use of animal feed material was known
to be technically feasible and economi-
cally attractive. Experiments were  de-
signed to further define this product as a
feed ingredient and to explore the cause
of depressed performance at  higher
levels of supplementation.
  Such experiments were designed to
evaluate  the biological availability of
phosphorus, pigmenting  value, and
protein quality of citrus sludge in poultry
feeds. The interaction between Vitamin
D3 and sludge was also examined as a
possible explanation for the depressed
performance seen at  higher levels of
supplementation. It has been theorized
that the high mineral content in sludge
catalyzed the destruction of Vitamin D3.
  These experiments were developed to
demonstrate the use of citrus sludge as
a feed ingredient to at least partially
offset both  dewatering and disposal
costs. The addition of citrus sludge to
the diet of broilers  did not affect the
palatability or texture of the meat. A
significant improvement in egg yolk
color  was also  observed when citrus
sludge was fed to hens. The flavors of
both the yolk  and albumen were not
affected.

Highlights of  Project
Findings

Metabolizable Energy of
Citrus Sludge
  Day-old chicks  were fed several diets
of citrus sludge substituted for an equal
amount of sucrose. During the third and
fourth week, representative samples of
excreta were  collected for analysis.
Moisture (percent dry  matter), gross
energy (expressed as Kcal/g), and
nitrogen and chromic oxide contents of
the various diets and excreta were
obtained to calculate the metabolizable
energy of citrus sludge.
  The  metabolizable energy of sludge
decreased  numerically  as  the sludge
substitution   level in  the diet was
increased (Table  3). Excessive supple-
ments of sludge (16 percent and above)
were found to  have a laxative effect on
the birds. This  may account for some of
the decrease in utilization of sludge.

Citrus Sludge Protein Assay
  A graded series of protein levels were
fed  to day-old  chicks for a  three-week
period in an attempt to assay the protein
quality of citrus  sludge. The negative
effect of sludge was reflected in its feed
intake efficiency. In the corn-soy diets,
as the protein level increased, feed
intake improved significantly. Conversely,
as  the percentage  of sludge was
increased, its  utilization decreased or
became masked, and a  depressing
effect occurred at the highest level of
supplementation  (Table  4).

Citrus Sludge-Vitamin Da
Interaction
  In two experiments, a graded series of
Vitamin Da and citrus sludge levels were
added to a corn-soy basal diet and fedto
day-old chicks  for a three-week period.
Tibia  ash,  phosphorus, and  calcium
values were obtained.  There  was no
significant difference between the
levels of sludge for values of calcium in
the first experiment, nor for phosphorus
or ash in both experiments. However,
there was an unexplainable increase in
calcium at the  five percent sludge level
in the second experiment. As the level of
Vitamin Da was  increased, ash values
increased numerically.
  There was  significant  interaction
between sludge and Vitamin Da. As the
level of Vitamin D3 was increased, feed
 Table3.    Metabolizable Energy Per
           Kilogram of Citrus Sludge

  Dietary Sludge Metabolizable Energy
  Level {percent)       (Kcal/kg)
2
3
4
6
8
16
3.368
2.411
1.813
1,360
1,404
836
intake increased. Sludge had no impact
on feed intake. In summary, the depressing
effect that citrus sludge had  on body
weight did not appear to improve with
the addition of Vitamin Da to the diet of
chicks.  Therefore,  the  cause of the
depression of body  weight by citrus
sludge was probably not  due to the
destruction of Vitamin Da by the level of
iron in the sludge, as has been previously
suspected. This lack of interaction was
also expressed in ash values

Phosphorus Assay of Citrus
Sludge
  Duplicate experiments were conducted
to evaluate the biological availability of
phosphorus contained in citrus sludge.
A modified corn-soy basal diet was
employed with  variable amounts of
citrus sludge in  which monosodium
phosphate, calcium carbonate, sucrose,
and filler  were added. Day-old  chicks
received each dietary treatment for a 3-
week  period. The addition of supple-
mental phosphorus  had no significant
influence in terms  of  body  weight.
However,  increasing calcium  levels
supported  some  numerical improve-
ments  in  body  weights.  All sludge
treatments had  weights significantly
less than the modified  corn-soy diet
which  had been supplemented with
phosphorus (from monosodium  phos-
phate). Calcium levels did not affect tibia
ash values from sludge treatments. The
failure  of sludge treatment to produce
better results may have been due to the
reduced  feed intake associated with
those treatments. This reduction was
felt to be  due to the large amounts of
sludge required  to add phosphorus to
the test.

Evaluation of Citrus Sludge
as an Effective Egg Yolk
Pigmenting Ingredient
  The purpose of this experiment was to
evaluate  the egg  yolk pigmenting
efficiency  of  waste citrus activated
sludge. Commercial hens were fed  a

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 Table 4.    Protein Assay Chick Performance* (3 Weeks)
    % Crude Protein
Body Weight (g)
Corn-Soy
16
18
20
22
16
16
16
Sludge
0
0
0
0
2
4
6
Exp. 1
364e
508b
549*
538*
479C
41 C?
302'
Exp. 2
404"
485*
536a
545a
45Sbc
430ca
289e
Feed Intake]
to)
33.6°
36.4ab
36. 8*"
35.2tic
37.3ab
38. 1a
30.5"
Feed Efficiency f
(g feed/g body wt)
1.80C
1.53s
1.421
1.361
1.70"
1.91h
2.24a
 * Means without common letters are significantly different according to Duncan's mufti
 pie range test (<0.05).
 t Values are a combination of two experiments.
white corn, basal, xanthophyll-free diet
for 15 days to deplete the yolk of all
pigment. The  yellow corn diets were
formulated to contain the same amount
of xanthophyll as each of the citrus
sludge  diets.  After  21 days on  the
experimental  diet,  the  eggs were
collected for analysis.
  The  data  indicated that egg yolks
produced by hens fed citrus sludge were
the same color as those produced by
hens fed yellow corn at equivalent
xanthophyll  levels. However, the  egg
yolks produced by hens fed citrus sludge
did not have as much pigment as those
produced by hens fed yellow corn. This
was probably a result of xanthophylls in
yellow corn  being  more  biologically
available. Citrus sludge would be a
suitable  pigmenting agent  for eggs
produced for  home  consumption.  It
would not be as suitable as yellow corn
in eggs produced for commercial
processes (e.g., mayonnaise,  noodles),
since there is less pigment in the yolks;
this pigment would be diluted, resulting
in processed egg products with less
than the desirable color.

Evaluation of Citrus Sludge
as an Effective Broiler Skin
Pigmenting Ingredient
  This  experiment was designed  to
examine  the influence of citrus sludge
on broiler pigmentation. Day-old chicks
were fed a white corn, xanthophyll-free
diet for a period of four weeks. Beginning
the fifth week, they were fed one of nine
experimental diets for three weeks.
  Pigmentation values  of  both shank
and  skin samples from  broilers  fed
yellow  corn  were significantly higher
than those fed the corresponding citrus
sludge diet. The feeding of citrus sludge
resulted in the same hue and darkness
of both the shank and skin samples as
did the  feeding  of  yellow  corn  at
                comparable xanthophyll  levels.  Pig-
                mentation values indicated  that the
                xanthophyll in yellow corn was slightly
                more  available than  that in  citrus
                sludge.

                Conclusions and
                Recommendations
                 Satisfactory conclusions could not be
                reached.  The chemical and  physical
                nature of  waste citrus activated sludge
                made  it  a  very  difficult product to
                evaluate in  the feeding trials. Several
                experiments did show that: high mineral
                content in citrus sludge was not an
                inhibitor of  Vitamin D3 efficiency, and
                citrus  sludge  possessed  pigmentation
                values approximately  equal to yellow
                corn in broiler skin and shanks and egg
                yolks.
                 It  is  doubtful  that  a citrus sludge
                similar to the one employed  in these
                experiments would  be feasible  in the
                commercial  poultry industry because of
                poor  nutrient content  and as-yet-
                unidentified depressive agents.  The
                identification  of  the agents  that are
                reducing bird performance is necessary.
                Further work is also needed to improve
                the methods of concentrating, drying,
                and handling the product to improve its
                nutrient content.
                                                                                     U 8 GOVERNMENT PRINTING OFFICE- 1M2 - 559 -017 /084Z

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      Bobby L Damron. Douglas M. Janky, Robert H. Harms, andMarkF. Hall are with
        the University of Florida, Gainesville, FL 32611.
      Kenneth A. Dostal and Harold W. Thompson are the EPA Project Officers
        (see below).
      The complete report, entitled "Evaluation of Waste Citrus Activated Sludge in
        Poultry Feeds," (Order No. PB 82-259 904; Cost: $7.50, subject to change)
        will be available only from:
              National Technical Information Service
              5285 Port Royal Road
              Springfield, VA 22161
              Telephone: 703-487-4650
      The EPA Project Officers can be contacted at:
              Industrial Environmental Research Laboratory
              U.S. Environmental Protection Agency
              Cincinnati, OH 45268
United States
Environmental Protection
Agency
Center for Environmental Research
Information
Cincinnati OH 45268
Postage and
Fees Paid
Environmental
Protection
Agency
EPA 335
Official Business
Penalty for Private Use $300

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