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’ BINDING BY

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”UK HINDU" l .
L! V Mn a

  

TAILH‘M

ABSTRACT

ORGAN WEIGHTS OF SINGLE COMB WHITE
LEGHORN COCKERELS FED PBB

BY
Edward H. Kowaleski Jr.

The recent accidental feeding of livestock and poultry
with.polybrominated biphenyl (PBB) has caused widespread
concern as to the exact effects of this chemical on exposed
animals. This study was undertaken to determine the effects
of feeding PBB to Single Comb White Leghorn cockerels. The
parameters studied were feed intake, body weight and various
organ weights; heart, liver, thyroid, testes, comb and spleen.

Symptoms observed after chronic administration (0-250
ppm) of PBB to 180 chicks and then to 210 chicks in a pair-
fed study (O-ZOOppm) included 1) depressed body weight as
a result of decreased feed intake, 2) decreased comb,
testes and spleen weights, 3) decreased heart weight as a
result of decreased feed intake, h) increased liver and thy-

roid weight.

ORGAN WEIGHTS 0F SINGLE COMB WHITE
LEGHORN COOKERELS FED PBB
By
Edward H. Kowaleski Jr.

A THESIS

Submitted to
Michigan State University
in partial fulfillment of the requirements
for the degree of

MASTER OF SCIENCE
Department of Poultry Science
1976

ACKNOWLEDGEMENTS

The author wishes to express his appreciation to his
adviser, Dr. Robert K. Ringer, for his unending guidance
and patience throughout the Master's program. The assis-
tance of Dr. Donald Polin and Dr. Lawrence Dawson in the
preparation and critical review of this thesis is greatly
appreciated. Special thanks are also extended to Dr. Theo
Coleman for his technical comments of thesis preparation.

Sincere gratitude is also given to Dr. Howard Zindel,
chairman of the Department of Poultry Science, for his help
and understanding.

Thanks go to the many graduate students in the Depart-
ment who helped with this study, especially Ms. Melinda Neff.
The excellent, time-consuming technical work of Ms. Maggie
Smith and the photOgraphic expertise of Mr. Sulo Hulkonen
is also acknowledged.

Finally, the author expresses his deepest appreciation
to his wife, Patti for understanding, patience and a good-
deal of typing, and to my family who have encouraged me

throughout.

ii

TABLE OF CONTENTS

Page
LIST OF TABLES O O O O O O O O O O O O O O O O O O O 0 O O O O O 0 O O O O O O O O O O O O O O 0 iv
LIST OF FIGURES O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O I O O O O O

<
P.

INTRODUCTION...........................................
OBJECTIVES.............................................
REVIEW OF THE LITERATURE...............................
Polychlorinated Biphenyls.........................
MATERIALS AND METHODS..................................
Experiment l......................................
Preparation of Feed...............................
Pair-Fed Experiment (Experiment 2)................

organSOOOOOOOOOOIOOOO000............OOOOOCOOOOOOOO

BOdy weight.‘......OOOOOOOOOOO......OOOOOOOOOO....

\OKOCIDCfiCthh-bbJNF-J

StatiStical AnalyseSOOOOOOOOOOO......OOOOCOOOOOOOO

RESULTS AND DISCUSSIONOOOOOOCOOOOO......OOOOOOOOOOOOOOO

F’ F1
c> C)

A. Experiment 1. - BOdy weightOCOOOOOOOOO00......

...:
O

01“ng YveightSOOOOO00.0.0000.........OOOOOOOOOO

W
O\

B. Experiment 2 (Pair-Fed Experiment) Body Weight

.;s
H

organ weightSOOOOOOOOOOOO......OOOOOOOOOOOOOOC

GENERAL DISCUSSION.0.000.000.0000.......OOOOOOCOOOOOOOO

«a -o
a; 0\

SUlvflVIARY AND CONCLUSIONS...O00............OOOOOOOOOOOOOO

\1
KO

BIBLIOGRAPI-IY......OOOOOOOOOOOO00.0.0000.........OOOOOOO

iii

TABLE

1.
2.

10.

ll.

12.

13.

LIST OF TABLES

Chi-Ck Starter rationOOOOO......OOOOOOOOOOOCOOOOO

Body and heart weights of SCWL cockerels fed
rations containing PBBOOO......OOOOOOOOOOOOOOOO.

Average weights of SCWL cockerels in grams fed
rations containing PBBOOOO...OOOOOOOOOOOOOOOOOOO

Average weight gain and feed consumption of
SCWL cockerels fed rations containing PBB.......

Liver and spleen weights of SCWL cockerels fed
rations containing PBBOOOOOOOOOOOOOOOOOOOOOOOOO.

Testes and thyroid weights of SCWL cockerels fed
rations containing PBBOOOOOOOOOOOOOOOOOOOOOOOOO.

Comb weights of SCWL cockerels fed rations con-
taining PBBOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOO

Pair-fed study of body and heart weights of SCWL
cockerels fed rations containing PBB............

Pair-fed study of feed consumption of SCWL cock-
erels fed rations containing PBB................

Pair—fed study of average SCWL cockerels body
weights (on a weekly basis) fed rations contain-

ing PBBOOOOOOOOOOOOOOOOOOOO......OOOOOOOOOOOOOOO

Pair-fed study of average SCWL cockerels body
weight gain (on a weekly basis)fed rations con-
taining PBBOOOOOOOOOOOOOOOOOOOOOOOOOOOOOO0.0....

Pair-fed study of liver and thyroid weights of
SCWL cockerels fed rations containing PBB.......

Pair-fed study of spleen weights of SCWL cock-
erels fed rations containing PBB................

iv

Page

13

14

15

27

32

33

44

45

46

47

57

58

LIST OF TABLES-- Continued
TABLE Page

14. Pair-fed study of testes and comb weights of
SCWL cockerels fed rations containing PBB....... 71

FIGURE
1.

10.

ll.

12.

l3.

14.

LIST OF FIGURES

Page

Body weight of SCWL cockerels fed rations
containing PBBOOOOOOOOOOOO00.0000000000000000.. 11

Heart weight of SCWL cockerels fed rations
containing PBBOOOOOOOOOOOOOOOOOOOOO00.000.00.00 l6

Liver weight of SCWL cockerels fed rations
containing PBBOOOOOOOOOOOOOOOOO0.00000000000000 18

Liver (g/lOOg bw) weight of SCWL cockerels fed
rations containing PBBOOOOOOOOOOCCCCQCOOO0.0... 20

Spleen weight of SCWL cockerels fed rations
containing PBBO00.00000........OOOOOOOOOOOOOOO. 23

Spleen (mg/100g bw) weight of SCWL cockerels
fed rations containing PBB..................... 25

Testes weight of SCWL cockerels fed rations
containing PBBOOOOOOOOOOOO...OOOOOOOOOOOOOOOOO. 28

Comb weight of SCWL cockerels fed rations con—
taining PBBOOOI.......OOOOOOOOOOOOOOOOO0.0...O. 3O

Comb (mg/100g bw) weight of SCWL cockerels fed
rations containing PBBOOOOCOOOOCOO......OOOCOCO 34

Thyroid weight of SCWL cockerels fed rations
containing PBBOOOOOOOOOOO0.00000000000000000000 37

Thyroid (mg/100g bw) weight of SCWL cockerels
fed rations containing PBB..................... 39

Body weight of SCWL (pair—fed) cockerels fed
rations containing PBB......................... 42

Heart weight of SCWL (pair-fed) cockerels fed
rations containing PBB............00......00000 48

Liver weight of SCWL (pair-fed) cockerels fed
rations containing PBBOOCOOOOOOO......OOOOCOOO. 51
vi

LIST OF FIGURES-- Continued

FIGURE

15.

16.

17.

18.

19.

20.

21.

22.

23.

Liver weight (g/lOOg bw) of SCWL cockerels fed
rations (pair-fed) containing PBB...............

Spleen weight of SCWL (pair-fed) cockerels fed
rations containing PBB...‘.................COOOO

Spleen weight (mg/100 bw) of SCWL cockerels
fed rations (pair-fed containing PBB...........

Testes weight of SCWL (pair-fed) cockerels fed
rations containing PBBCOOCOOCOCUOOOOCOCC0.00....

Comb weight of SCWL (pair—fed) cockerels fed
rations containing PBBCOOOCCOOOOC......OCOOCCOC.

Testes weight (mg/100g bw) of SCWL cockerels fed
rations (pair-fed) containing PBB...............

Comb weight (mg/100g bw) of SCWL cockerels fed
ratiOnS (pair-fed) containing PBBooooooooooooooo

Thyroid weight of SCWL (pair-fed) cockerels_fed
rations containing PBBCCOOOC......COOOOOOOOOCOOO

Thyroid weight (mg/100g bw) of SCWL cockerels
fed rations (pair-fed) containing PBB...........

vii

Page

53

55

61

63

65

67

69

72

74

INTRODUCTION

The dispersion of chemicals into the environment
has become an issue of primary concern. The problem can
be especially acute as when an accidental contamination
occurs as it did in Michigan in 1973. The fire retardant,
polybrominated biphenyl (PBB), was inadvertantly mixed into
livestock and poultry feeds, resulting in considerable eco-
nomic loss and concern for human contamination.

PBB is used in.the thermOplastics industry mainly as a
component of the housings of business machines and in indust-
rial and electric equipment. It is a relatively new compound
and thus has received very little toxicological research.

The present study is a report of the effects of PBB on
body weight, various organ weights and on feed consumption
in the Single Comb White Leghorn cockerel, when administered

continuously in the feed.

Objectives

This research was conducted to measure the effects of
feeding polybrominated biphenyl to Single Comb White Leghorn
cockerels.

Specific objectives were;

1. To determine the effect on body weight gains;

2. To determine the effect on feed consumption;

3. To determine the effect on weights of various organs

of the body including the heart, liver, spleen,
testes, comb and thyroid.

REVIEW OF LITERATURE

Toxicological effects of polybrominated biphenyls (PBB)1
have not been studied in animals as extensively as the chemie
cally related polychlorinated biphenyls (PCB) but with the
recent accidental exposure of livestock and poultry to P38
in the state of Michigan it is evident that research is
greatly needed.

P88 is an industrial compound used.main1y in the thermo-
plastics industry to reduce flammability. It is a.mixture
of brominated biphenyls with the average number of bromine
atoms being six per biphenyl molecule. The mixture is as

follows;
Tetrabromobiphenyl 2.0%
Pentabromobiphenyl 10.6%
"Hexabromobiphenyl 62.8%
Heptabromobiphenyl 13.8%

Other‘bromobiphenyls ll.M%

It is relatively inert chemically but a study of the
photolytic stability of octabromobiphenyl indicates ultra-
violet light readily degrades this material under conditions
where PCBs are not degraded (Kerst, 197k). The use of P33
has been restricted, ideally, to end-use products that are
not exposed to either feed or food. It is also not used in

flame retarding fabrics where human exposure might occur

1Amulticomponent mixture produced by The Michigan Chemical

00., Ste Louis, M1. 3

(Kerst, 197k). PBB is a relatively new compound with total
production estimated at about eleven.million pounds up to
197k (Kerst, 197a).

The literature on PBB metabolism in biological systems
is scanty, with avian species receiving very little attention.
Lillie gt.gl&_ (197h) included one level of PBB (BF-6) in
their study on reproduction in hens and demonstrated signifi-
cantly reduced feed intake per hen-day. Egg production was
also significantly lower while adult body weight gain, liva-
bility, egg weight, egg shell thickness and fertility were
not effected at the 20 ppm administered. Source progeny per-
formance tests were also reported showing significantly re-
duced three week body weight gains in chicks from.hens that
had received 20 ppm PBB in their diets. Cecil 2;; Q; (1975)
working with Japanese quail reported reduced phenobarbital
sleeping time with a single oral injection of 300 ppm.PBB.

No data are available on the effects of PBB feeding on organ
weights of birds.

Liver enlargement with P835 has been reported by Aftosmis
9335 (1972) and by Norris 2331; (1973) both working with

rats.

Polxchlorinated biphenyls

The chemical and physical characteristics of PBBs and
PCBs are similar. Therefore similar results would.be expected
from the ingestion of the chemicals. PCBs have been widely

used in industry in electrical capacitors and in electrical

transformers as well as in heat-exchange fluids. PCB's have
been used by the food industry in pasteurization equipment.
It was therefore not surprising that this compound found it's
way into fishmeal that was to be used in the poultry industry
(Harris and Rose, 1971) as feed. The symptoms observed were
those similar to chick edema disease (McCune e£_§;;, 1965)
including varying degrees of liver degeneration, small
spleens and muscle incoordination.

Various studies undertaken in chickens have shown in-
creased liver weights with PCB ingestion (McCune et_§;&,
1962; Flick g§_§;&, 1965; Platonow and Funnell, 1971).
Splenic atrophy has been reported by Flick 22.91, (1965),
while Platonow and Funnell (1971) have indicated that testes
and comb size are reduced in chickens given 150 to 250 ppm
PCB. Increased thyroid size and weight was reported by
Jefferies and Parslow (1972) in the young gull following
PCB administration. Rehfeld e§_§;, (1971) using a PCB
(Aroclor 21h8) at 50 ppm observed in the chick: depressed
weight gain; edema; hydropericardial fluid; depression of
secondary sexual characteristics; and an increase in liver
size as a percentage of body weight. Iturri (l97h) also work-
ing with the chick demonstrated decreased heart, spleen, testes
and comb weights while showing increased liver and thyroid
weights with varying levels of PCB.

Materials and Methods

Experiment 1.
One-day-old commercial (DeKalb Strain) Single Comb White

Leghorn (SCWL) cockerels (180) were reared in conventional
electrically heated battery brooders adjusted to 35°C, with
raised wire floors. They were randomly divided into six
groups of thirty birds each, weighed individually, and wing-
banded.

The control group was treated like all others except
that they did not receive any PBB in their feed. The five
treatment groups received feed with varying amounts of BBB.
The amounts added were as follows: 50 ppm, 100 ppm, 150 ppm,
200 ppm, 250 ppm.

The experiment was conducted for a period of h weeks.
Feed and water were provided ag_libitum. Body weights and
feed consumption of the chicks were recorded weekly. To keep
the wastage of feed to a minimum the feeders were half-filled

every day.

Preparation of Feed

Diets containing the different levels of PBB were pre-
pared by adding the respective amount of PBB to a chick start-
er ration (Table 1).

The chemical was ground to a fine powder with a mortar
and pestle and then weighed on a Mettler HlOTw (Sargent Welch
Scientific Co.) balance. Approximately five hundred grams of

the starter ration was then screened (U.S. Standard Sieve
6

7
Table l. Chick starter ration.

 

 

 

Ingredients Lbs per ton
Corn 1239
Soybean meal, h9% not
Alfalfa meal, 17% 50
Meat 3: bone meal, 50% 60
Fish.meal, 60% 60
Whey, dried no
Oats 100
Salt 5
Limestone, ground 15
Dicalcium phosphate 15
Premix (5003) 10
Additives 2

2000

 

8
Series No. 20) three times. This powder was then mixed

with the respective PBB. This premix was combined and mix-
ed thoroughly with the basal ration using a mechanical
rotary feed mixer. The same procedure was utilized for the
control group except that PBB was not added. All rations
were stored in 10 gallon metal containers (Reeves, Dover,

Ohio) prior to and during their use.

Pair-Fed Experiment
The possibility existed that feed consumption might alter

some of the parameters being analyzed. To study this effect,
seven groups of thirty SCWL chicks per group were utilized.
The experimental groups were given a known amount of feed
where PBB had been incorporated and treated as described pre-
viously. The levels in the diets were: 50 ppm, 100 ppm, 200
ppm. Feed consumption from these groups was measured on al-
ternate days using a Toledo model 3710 (Toledo Scale Corp.)
scale, and an equal amount of the basal ration without PBB
was given to the pair-fed group. In this manner the two-day
lag allowed the pair-fed group the same amount of feed con-
sumed by the experimental group. This procedure was again
followed for a four week period. The seventh group or thirty
chicks acted as the control and was fed the chick starter

ration ad libitum.

Organs
In both experiments, at the end of the four week periods

the birds were sacrificed and various organs were dissected

9.
free of fat and surrounding tissue and weighed on a Mettler

P1210 (E. H. Sargent and Co.) balance to the nearest 0.1
grame The organs were kept moist on wet paper towels be-
fore the actual weighing. The organs collected were heart,
liver, thyroids, testes, comb and spleen. To insure great-
er accuracy, thyroids were weighed on a Roller-Smith pre-
cision balance (Fisher Scientific 00., Pittsburgh, Pennsy-
lvania) to the nearest 0.1 mg.

Body weight
The chicks were weighed at the initiation of the experi-

ments and for the next four weeks at weekly intervals using
a Mettler P1210 (E. H. Sargent Co.) balance. They were

weighed to the nearest 0.1 gram.

Statistical Analyses
Statistical analyses of the data Obtained were accomp-

lished by analyses of variance. Parameters showing signifi-
cant effects were investigated further by Dunnet's Test.

RESULTS AND DISCUSSION

A. Experiment.1.

Body Weight
Body weight was significantly reduced by PBB at the 100

ppm dosage as is shown in Figure l and in Table 2. It is
apparent from these data that with increasing concentrations
of PBB there is a concomitant decrease in body weight. At
the end of a two week period the birds in the 200 and 250
ppm groups were experiencing increased mortality and were
very emaciated and lethargic. The depression in body weight
observed is related to a reduction in feed intake. As illus-
trated in Table h, by the fourth week the controls were eat-
ing 39.2 g/chick/day but the 250 ppm.group was consuming only
23.3 g/chick/day, some h0% less. Similar depressions in
growth have been reported with P033 by Flick 2:; _a__l_,_ (1965).
Organ'Weights

The various organ weights are presented in Tables 2,5,6
and 7 and Figures 2 through 11. Mean heart weights were
significantly reduced at the 50 ppm.and above dosage. From
Fig. 2 it is estimated that a significant decrease in heart
weight would be obtained from a dosage between 25 and 30 ppm.
Iturri (1971;) found a significant decrease in heart weight
when feeding PCB 125k but with no other PCB.

The absolute mean liver weights were not significantly
different throughout the range of dosages; however, the
relative liver weight increased over controls at all levels.
Since no change in actual organ weight was seen, the relative

increase could be accounted for by the decrease in body

10

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13

Table 2. Bow and heart weights of SCWL cockerels fed

rations containing PBB.

 

 

 

PBB Body wt. Heart wt. Heart
Level (ppm)a (gm) (gm) (% b.wt)

o 331.7:s.u67b 2.0919.058 0.63:9.016
(30) (30) (30)

so 311.215.09h 1.75:9.ohe 0.56:9.011
(30) (30) (30)

100 293.3+8.553 1.62:9.050 0.53:9.013
(28) (27) (27)

150 269.11,.756 l. 7:9.036 0.55:9.013
(30 30) (30)

200 2 .9+9.108 1.53+0.067 0.6 +0.031
ul(2€) (26) (%€)

250 209.3+9.953 1.37+0.089 0.66+0.033
(25) (25) (25)

 

a‘Rations fed up to I). weeks

bData are reported as Mean i. Standard Error
(No. of birds)

Table 3 0

1h

Average weights of SCWL cockerels in grams fed
rations containing PBB.

 

 

 

LeveIBIppm)a Dgy Dgy IE? 22y gay
0 38.2 78.2 lh8.5 2h0.3 331.7
50 36.9 78.6 lh3.h 226.5 311.2
100 37.7 7u.h 129.7 210.6 292.h
150 36.5 72.2 123.h 197.2 269.1
200 36.7 68.6 111.0 175.7 2u1.9
250 36.9 66.6 107.3 160.2 209.3

 

aRations fed.up to u weeks

Table h. Average weight gain and feed consumption of SCWL
cockerels fed rations containing PBB.

 

 

 

‘, __Avg. Wt. Gain -g
Days Days ays

 

 

 

 

LeveIBIppm) 0-7 8-1h 15-21 22228
O u0.5 70.7 91.8 91.9
50 hl.6 6h.8 83.7 8h.8
100 36.7 55.3 80.5 81.8
150 35.7 51.2 73.8 71.9
200 31.9 h1.2 6h.7 66.2
250 29.6 39.8 52.9 51.5
Feed Consumption - chhick/dqz

3597's 3531? 35331 33338

O 10.15 20.1 29.6 39.2
50 lO.h9 19.2 27.1 35.6
100 9.75 18.2 211.5 33.5
150 9.66 13.1 22.9 3lch
200 8.8h 13.5 20.6 25.9
250 8.93 1h.3 19.6 23.3

 

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weight. Similar results with mean liver weights were re-
ported (Rehfeld gt_§1., 1971; Platonow and Funnell, 1971)
with PCBs in chicks, however other investigators have found
increased weights of the liver with increasing concentrations
of PCB (Flick _e_1;_ _a_1_., 1965; Grant 21 21.; 1971). Iturri
(1974) also demonstrated increased liver weights in chicks
with PCB 1242 and 1260.

Spleen weights were significantly reduced at levels 2
100 ppm. As a percentage of body weight they also display-
ed a decreasing trend. This seems to be consistent with the
data for PCBs in chickens (Flick 23.21,, 1963; Iturri, 1974)
and in rats (Grant et_§1,, 1971). No explanation has been
given for the decreased size of the spleen and further histo-
logical studies will no doubt be extremely helpful.

Comb and testes weights will be considered t0gether(Fig~
ures 7,8, and 9, Tables 6 and 7) because of their obvious re-
lationship. Comb weight was significantly reduced at all
levels tested therefore the dosage at which PBB has a signif-
icant effect on the comb would be approximately 20 to 25 ppm.
It is interesting to note that although comb weight was sig-
nificantly reduced at 50 ppm, testes weight was not depressed
until the 200 ppm level was utilized. As can be seen in Fig.
7 the birds fed the 50 and 100 ppm diet have testes actually
weighing more. This could be a drug effect caused perhaps
by increasing the amount of adipose tissue in the testes, or

it could be experimental error. Whichever the case, the

testosterone is not being produced and/or not having a phys-

23

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Table 5. Liver and spleen weights of SCWL cockerels fed
rations containing PBB.

PBB a Liver wt. Liver Spleen wt. 8 leen

Level (ppm) (gm) (% b.wt) (mgo) ( b.wt)

o 7.99:9.27ub 2. :p.oau 519_32. 7 155+9. 8
(30) 30) (3o) (30)

so 9.29:9.29u 2.99:9.102 u72+25.9 815.8
(30) (30) (3o) 30)

100 9.17+o.293 3.12:9.09u 7+16. 2 11&_5. 8
(57) (27) 27) (27)

150 9.99:9.330 3.72+o.107 286+1u.3 106+u. 3
(3o) (36) (30) (30)

200 9.9 +0.h85 h.03+0.108 213+15.9 86+ 07
?§6) (26) (263 (26%

250 9.03+o.513 n.26+o.133 18u+15.2 861g.7
(in) (2H) (23) (23)

 

aRations fed up to a weeks

b

Data are reported as Mean + Standard Error
(No. of birds)

28

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Table 6. Testes and thyroid weights of SCWL cockerels fed
rations containing PBB.

PBB Testes wt. Testes Thyroid wt. Thyroid

Level (ppm)a (mg) (% b.wt) (mg) (% mm)

o 76. 9 .ob 23.u+1.o 17.3+o.7 54910.20
(35% (36) (3'6) (30)

So 79.813.o 25. 7+0. 9 20. 2+1. 0 6. 619.29
(30) (30) (30) 30)

100 81.7+u.8 27.S+1.u 23.5114; 7.9h;0.h.2
(2‘5) (26’) (27) (27)

150 71.0 _3 8 26. 3_+_1. 2 25.6115 9.53:0.52
(30) (30) (30) (30)

200 66.1+h.h 27.0+1.2 25.6+1.6 10.7h+0.52
(26') (26') (23) (23)

250 h8. S+h .1 22.6+1.S 22.8+l.5 10.7hao.50
(21?) (2E) (21?) (2E)

 

aRations fed.up to h weeks

hData are reported as Mean + Standard Error

(No. of‘birds)

33
Table 7. Comb weights of SCWL cockerels fed rations contain-

 

 

 

ing PBB.
PBB Comb wt. Comb
Level (Ppm)a (mg) (% b.wt)
o 1061+78. ob 32%+25. .3
(30)
so 656_561 209;16.3
(30) (30)
100 M71385 11.81102.
(27) (27)
150 379126.? 13639. 6
(30) (30)
200 258+23. 8 10%+8. .2
(26) 26)
250 176+2h.5 80+9.2
(5h) (5h)

 

aRations fed up to h weeks

bData are reported as Mean + Standard Error

(No. of birds)

34

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36
iological effect on the comb. Platonow and Funnell (1971)
in a 13-week study demonstrated similar weights in testes
at the end of a six week period between controls and PCB
treated cockerels but by nine weeks there was a significant
decrease. They also noted a decreased comb growth appearing
before a similar decrease in size of the testes.

Jefferies and Parslow (1972) demonstrated increased thy-
roid weights in the gull with the administration of PCB. They
described the condition as being similar to that of simple
goiter, and suggested interference with the iodine trap mech—
anism or decreased thyroid stimulating hormone quantities.

In this study thyroid weight was significantly increased at
all levels of PBB above 50 ppm, Fig. 10. Interestingly

there appeared to be a leveling off of thyroid weight between
150 and 200 ppm PBB and a decrease below the 200 ppm weight,
by the group fed 250 ppm. This indicated that the toxicity
was too great at the 250 ppm level and that some form of de-
generation was taking place overcoming the stimulatory effect
seen at lower levels. Plasma thyroxine concentration in PCB
treated birds (Iturri, 1974) was significantly increased by
PCBs 1254 and 1260. Why the thyroxine concentration is aug-
mented and the mechanism by which the treated thyroids in-

crease in size is not clear.

B. W)
Body Weight

The object of a pair-fed study is to eliminate the in-

37

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fluence that increased or decreased feed consumption might
have on experimental results. In Experiment 1 PBB depressed
feed consumption; therefore a pair-feeding study was conduct-
ed to test the effects of the drug pg; EE- Since the treated
and the pair-fed group received equal amounts of feed it was
presumed the effects seen were due only to the action of the
drug. As seen in Fig. 12 it would appear that PBB acted as

a growth stimulant at low dietary levels. This is in con-
flict with a pair-fed study with PCBs by Iturri (1974) who
demonstrated no significant difference between pair-fed and
treated birds. It was felt that the chicks that received 50
and 100 ppm pair-fed levels were far less effecient than the
controls. The pair-fed 50 group consumed 99.5% of the con-
trol intake (Table 9) Yet were some 40 grams lighter in body
weight at the termination of the experiment. As will be
pointed out later, Fig. 16, the spleens of this group were
greatly enlarged, indicating other types of problems. These
problems have precluded the use of the data as a percent of
body weight and these are included only for the sake of com-
pleteness. As seen in Table 11 the treated 50 ppm and 100
ppm PBB groups consistently outgained their pair-fed counter-
part on an average weight gain basis, so it would appear that

there was a problem from the beginning.

Organ Weights
Heart weights were significantly reduced in the 2 100

ppm groups. There were, however, no significant differences

within the respective groups, for this reason a single lepe

42

Figure 12. Body weight of SCWL (pair-fed) cockerels fed
rations containing PBB. Mean values + S. E. Y

= body weight (grams); X = PBB level ng/kg) in
the diet.

wl.

Body

43

 

Jaunt—

320

#— PBB

I — Pair-fed
3
280
26
240
220

 

50 100 200
PBB- mg /kg die!

44

Table 8. Pair-fed study of body and heart weights of SCWL
cockerels fed rations containing PBB.

 

 

“——

 

PBB a Body wt. Heart wt. Heart
Lovel jppm) (gml i (gm) (ngOOgjw)
o 32u.u+ .9b 2.11:9.07 0.65:9.02
(30 (30) (30)
Pair-Fed
o 280.7+u.u 2.07:9.07 0.74:9.02
(297 (29) (29)
£22.
50 309.1+ .7 1.99:9.07 0.6419.02
(29 (29) (29)
Pair-Fed
100 258.6+ .9 1.8710.0h 0.72:0.02
(3o (30) (30)
£22.
00 285.9+ .9 1.85+o.07 0.6u+o.02
(29 (28) (28‘)
Pair-Fed
"266' " 226.5+ .7 1.59:9.05 0.7019.02
(29 (29) (29)
EEE. 8
200 23o.7+ . 1.71+o.07 0.7h+o.03
(28 (28') (28’)

 

aRations fed up to h weeks

b

Data are reported as Mean_:_Standard‘Error

(No. of birds)

45

Table 9. Pair-fed study of feed consumption of SCWL cock-
erels fed rations containing PBB.

 

 

 

Feed Consum tion - chick da
PBB lst 2nd 3rd Eth

 

Level (ppm1_f Week ngk Week Week
0 9.8 17.8 25.8 32.9
Pair-Fed
0 10.2 18.0 25.1 32.6
PBB
50 10.2 18.0 25.1 32.6
Pair-Fed
100 10.1 17.3 23.0 29.1
PBB
100 10.1 17.3 23.0 29.1
Pair-Fed
200 9.9 15.9 20.7 21.8
PBB
200 9.9 15.9 20.7 21.8

 

46

 

 

 

 

 

Table 10. Pair-fed study of average SCWL cockerel body
weights (on a weekly basis) fed rations contain-
_ Avg. Chick wts. - R
PBB Day Day Day Day Day
Level (ppm) 0 7 1h 21 28
o h9.6 83.9 1h8.8 231.5 321.u
Pair-Fed
0 h8.0 73.0 132.7 205.5 280.7
218.8.
50 h7.7 82.9 1hh.l 223.0 309.1
Pair-Fed
100 u9.6 72.6 126.6 193.1 258.6
221.3.
100 u9.1 78.u 132.9 206.3 285.9
Pair-Fed
200 h9.6 70.5 117.5 176.0 226.5
PBB
"‘200 18.9 75.8 121.3 180.u 230.7

 

Table 11. Pair-fed study of average SCWL cockerel body -
weight gain (on a weekly basis) fed rations con-
taining PBB.

47

 

 

Avg. Wt, Gain -R

 

 

 

 

PBB pays Days Days Days
Level (ppm) o-7 8-lh 15-21 22-28
0 3h.3 6h.9 86.0 98.7
Pair-Fed
0 25.0 59.7 72.8 73.8
2.32:.
50 38.8 61.3 78.9 86.1
Pair-Fed
100 22.9 5h.0 66.5 65.5
213.12.
100 29.3 58.6 73.1 78.3
Pair-Fed
00 20.8 h7.0 59.8 h9.1
PBB
"280 26.9 85.5 59.0 u8.9

 

48

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50
is provided. The results indicate that the drug has no
effect on heart weight. The hearts from the PBB-treated
groups throughout appeared to be more flaccid and not as
thick-walled as the hearts of controls and on a size basis
they seemed to be equal to, or larger than the controls.

Mean liver weights were significantly increased at 2
50 ppm (Fig. 14), while-the pair—fed groups showed a de-
creasing trend with the pair-fed 200 group having signifi-
cantly lower liver weights. This demonstrates that PBB has
a definite effect on liver weight. This effect was potent-
iated as the level of PBB was increased as indicated by the
difference between the groups at the 200 level. On a rela-
tive basis the pair-fed birds show essentially no change with
starvation, the treated, on the other hand, increased dramati-
cally. Results from this pair-fed study is in agreement with
another pair-fed study (Iturri, 1974) in which PCB 1242 was
incorporated at 100 ppm. In Iturri's study the treated group
was not significantly different from the control, but there
was a significant decrease in the pair-fed liver weight.

This demonstrated that PCB maintained the liver weight while
it should have decreased.

Figure 16 depicts the results of PBB ingestion on mean
spleen weights. A significant difference was seen at 100 and
200 ppm PBB. This was in contrast to their unchanged pair-
fed groups. The pair—fed 50 group had a very high weight.
Considered with the decreased body weight of this group an

infection or some other disorder might be indicated. The

51

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Table 12. Pair-fed.study of liver and thyroid weights of

SCWL cockerels fed rations containing PBB.

 

 

 

PBB Liver wt. Liver Thyroid wt. Thyroid
£9221 (ppm) (2211 jig/100g bw) £55) 1355492333)
0 9.30:0.28b 2.87:0.06 16.8§+1.09 5.00:0.30
(30) (30) (207‘ (20) -
Pair-Fed
0 9.97:0.28 3.53:0.09 17.82+1.21 6.16:0.82
(29) (29) (201' (20)
EEQB 8 88 8S 6 6 56
0 11. 7:0.30 3. +0.07 21. +2.03 . 9:0.
(29) (2'9") (207 (20)
Pai Fed
“'15—— 8.90+0.28 38110.09 15.52+0.95 6.00:0.38
(35) 30) (201' (20)
a? 9+ 1. 8 69 9 s a u
00 13. 0. 0 . +0.10 23. 0+1.3 . 3:0. 7
1(28) (28) (201' 120)
Pair-Fed
__200 7.81_+_0.22 38510.09 13.31+0.85 5.85:0.38
(29) (29) (207 (20)
288, 86+ 55 S 8 6 8 6 8 88
200 13. 0. .9 +0.1 21. 2+1. 2 10. 2:0.
(28) (28) (207 (20)

 

aRations fed up to 8 weeks

bData are reported as Mean.;,Standard.Error
(No. of birds)

58

Table 13. Pair-fed study of spleen weights of SCWL cockerels
fed rations containing PBB.

 

 

PBB Spleen wt. Spleen

 

Leveljppmlg (g5) (mgfilOOg bw)
0 618. 93:33.8" 191. 5:10 . 2
(30) (30)
Pair-Fed
50 812.6:59.8 292.8:23.1
(29) (29)
Egg; 5 8 6 8 6 61 0 1
0 0 O + 2. l O + 200
(2'9) (29')
Pair-Fed
"‘"""""'100 599. 3181.1 231.1+18.2
(29) (29)
225% 398 S 23 5' 136 2 6 8
00 O + O O + O
(28') (28')
Pair-Fed
200 581.6+85 .0 236.8;18.7
(29) ' (29)
BEE. 58 8 6
200 2 .1+23.9 107.7+ .
(28') (28’)

 

aRations fed.up to 8 weeks

bData are reported as Mean.:_Standard.Error
(No. of birds)

59

exact reason for this increase is not evident. Utilizing

the 100 and 200 ppm groups a definite effect can still be
discerned by PBB administration on the spleen. A decrease

in relative spleen weights over pair-fed and controls is also
illustrated (Fig. 17).

As was shown in Fig. 7, there was first an increase in
weight of the testes at lower levels (Fig. 18) and then a
steady decrease to a significant level at 200 ppm. The drug
effect was already seen by 50 ppm and as the PBB level is in-
creased, more of an effect is seen from starvation as is dem-
onstrated by the converging lines. The comb effect elicited
in Experiment 1 (Fig. 8) was also evident in the group fed 50
ppm. It would appear then that this would be a consistent
effect of the drug. Similar results were found by using PCBs
in chickens (Platonow and Funnell, 1971). Testes and comb
weights on a percentage basis (Fig. 20 and 21) were also
shown to decrease with increasing concentrations of PBB, with
the maximum effect seen at 50 ppm.

Mean thyroid weights are interesting (Fig. 22). There
was a significant increase in thyroid weight in all PBB levels
utilized while the pair-fed displayed a downward trend, though
not statistically significant. The treated groups displayed
first an upward, then downWard trend (Fig. 10). This re-
search does not presume what might happen to thyroid weight
at a higher level of drug, 300 ppm for example, but Figures
10 and 22 indicate that there is some toxic effect occuring
at 200 to 250 ppm PBB. The relative thyroid weights (Fig. 23)

gave a misleading pattern for although at the 200 ppm level

60

the absolute weight was decreased, the relative weight show-

ed an increase.

61

Figure 17. Spleen weight (mg/100g bw) of SCWL cockerels fed
rations (pair-fed) containing PBB. Mean values
+ S. E.

62

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65

Figure 19. Comb weight of SCWL (pair-fed) cockerels fed
rations containi PBB. Mean values + S. E.‘Y
= comb weight (mg 3 X: PBB level (mg/kg) in the
diet; A indicates a non-significant change
(p<0005) e

66

 

  
   
   

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Table 18. Pair-fed study of testes and comb weights of SCWL
cockerels fed rations containing PBB.

 

 

 

PBB Testes wt. Testes Comb wt. Comb
Level (ppmja' (mg) (mg/100g bw) Lang (mgllOOg bw)
0 95.231837b 29.3811-29 1582.0+111.6 875.5133.8
(3o) (30) (3d? (30)
Pair-Fed 5 8 8 38. 8 S 8 8 S
0 9 .00+ . 0 23+1. 9 139 .7+10 . 00. 0_37.0
(29) (29) (297’ (29)
£22.
50 129.7218.79 81 95+2. 58 1080.6+108.8 350. 8_38. 3
(29) (29) (297’ (29)
Pair-Fed
“1fifif"' 75.5713.08 29.18+1.08 988w .6+62.6 382.8128.3
(29) (29) 307 (30)
£88, 8 8 85 S 5 8.
100 111.3 + 093 3 0 +1090 72 01+ 70 9. 9+1 2
(285 (28) (281'5 an (28)
Pair-Fed
"260"" 61. 01_3. 02 26. 88+1.28 709.2+ 5.8 310-9123-2
(29) (29) (29 (29)
255' 96 8 8 68 S 8 6
00 79. + .1 33. + .03 3 7.9+ 0. 1 9. 2+ .
(28) (285 (287' 8 (2811"7

 

aRations fed up to 8 weeks

b

Data are reported as Mean 1 Standard Error

72

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GENERAL DISCUSSION

Toxicity symptoms after oral administration of PBB were
similar to those described for PCBs by other workers. A
very evident symptom was decreased body weight. Comparing
PCB pair-fed work (Iturri, 1974) and the present PBB study
it was evident that the decreased feed intake was respon-
sible for the lower body weights and not an effect of the
drug pg£_§g, The drug could be unpalatable to the bird or
may act as a depressant on the appetite center in the hypo-
thalamus.

The heart weight was significantly reduced in both ex-
periments when PBB was fed. However, the pair-fed study de-
monstrated that the decrease was due to the lowered feed in—
take and not due to the drug directly. As was pointed out
previously, although there was no change in weight there
seemed to be morpholOgical changes in the heart of treated
birds. The hearts appeared to be thin-walled and flaccid.

The effects of PBB on the liver in Experiment 1 are mis-
leading on an absolute weight basis unless they are consid-
ered with the pair-fed study. This study demonstrated that
in a feed-reduction condition the liver would decrease in
size, but in Experiment 1 they remained the same and in the
pair-fed the mean weights increased. These results provide

evidence that PBB does cause definite changes in the liver.
76

77

Mean spleen weights in treated groups were decreased
in both experiments. Although the high spleen weight in the
pair-fed 50 group precludes making a definite statement, it
would appear from this and similiar studies with PCBs (Flick
21.El°: 1965; Iturri, 1974) that the drug does have an effect
on the spleen and that this is not the result of decreased
feed intake, but an action on the spleen itself or the imp
munological system.

Testes and comb weights show an interesting and consist-
ent pattern in both studies. Comb weight is significantly
reduced at the 50 ppm level but the testes weight at this
level is equal to or above the weights of testes of controls.
The effect on the comb is therefore elicited before a sim-
iliar effect is seen in the testes. Decreased secondary sex-
ual characteristics is indicative of interference in the test-
osterone production or utilization mechanism. The specifics
of these results are not clear.

The significant increase in weight of the thyroids, the
leveling off and the decrease in weight at the 250 ppm level
indicate a stimulatory action at lower levels. As the dos-
age in the feed is increased a toxic effect seems to be dem-
onstrated and finally a decreasing trend is noted. Similiar

results were seen in the pair-fed study.

SUMMARY AND CONCLUSIONS

Symptoms observed after chronic administration (0-250
ppm) of PBB to 180 chicks and then again to 210 chicks in a
pair-fed study (0-200 ppm) included 1) depressed body weight
as a result of decreased feed intake, 2) decreased comb,
testes and spleen weights, 3) decreased heart weight as a
result of decreased feed intake, 4) increased liver and thy-

roid weights.

78

BIBLIOGRAPHY

BIBLIOGRAPHY

Aftosmis, J. G., 23, al., 1972. The toxicology of brominated
biphenyls - oral toxicity and embryo toxicity. Paper
presented at the society of Toxicological Meeting,
Williamsburg, Virginia. As cited from A. F. Kerst,

1974.

Cecil, H. C., S. J. Harris and J. Bitman, 1975. Effects of
polychlorinated biphenyls and terphenyls and polybroms
inated biphenyls on pentobarbital sleeping times of
Japanese quail. Arch. Environ. Cont. Toxicol. 3:
183-191.

Dunnett, C. E., 1955. A multiple comparison procedure for
comparing several treatments with a control. J. Amer.
Stat. Assoc. 50: 1096-1121.

Flick, D. F., R. G. O'Dell and V. A. Childs, 1965. Studies
of chick edema disease. 3. Similarity of symptoms
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Grant, D. L., W. E. Philips and D. C. Villeneuve, 1971.
Metabolism of a polychlorinated biphenyl (Aroclor 1254)
mixture in the rat. Bull. Environ. Contam. Toxical.

6: 102-112.

Harris, J. R. and L. Rose, 1972. Toxicity of polychlorinated
biphenyls in poultry. J. Amer. Vet. Med. Assoc.,
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Iturri, S. J. 1974. The effects of various chlorinated hydro-
carbons on the cardiovascular physiology and hematology
of the domestic fowl. Ph,D. Thesis, Mich. S. Univ.

Jefferies, D. J. and J. L. Parslow, 1972. Effect of one
polychlorinated biphenyl on size and activity of the
gull thyroid. Bull. Environ. Contam. Toxicol. 8:
306-310. .

Kerst, A. F., 1974. Polybrominated biphenyls. Paper pre-
sented before the Michigan Environmental Review Board.

79

8O

Lillie, R. J., H. C. Cecil, J. Bitman and G. F. Fries, 1974.
Differences in response of caged White Leghorn layers
to various polychlorinated biphenyls (PCBs) in the diet.
Poultry Sci. 53: 726-732.

McCune, E. L., J. E. Savage and B. L. O'Dell, 1962. Hydro-
pericardium and ascites in chicks fed a chlorinated
hydrocarbon. Poultry Sci. 41: 295-299.

Norris, J. M., 23, 21., 1973. Toxicological and environ-
mental factors in the selection of decabromobiphenyl
oxide as a fire retardent chemical. Paper presented
to the Applied Polymer Symposium, No. 22 as cited
from A. F. Kerst, 1974.

Platonow, N. S. and H. S. Funnell, 1971. Anti-androgenic-
1ike effect of polychlorinated biphenyls in cockerels.

Rehfeld, B. M., R. L. Bradley, Jr. and M. L. Sunde, 1971.
Toxicity studies on polychlorinated biphenyls in the
chick. I. Toxicity and symptoms. Poultry Sci. 50:
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HICHIGQN STQTE UNIV. LIBRRR

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