FAC’EGPLS AFFECTERG. LEVER FAY AC€E§MULAT§OK
AM} LEVER EERﬁORERAGES ASSOCEAI’ED
‘6’ 2T3 FATTY LEVER “ HEMGRREAGIC
SYNDRWE ER LAYERS {IEECKENS

”(em '30? mam a; Db. D. , I
mam»: STATE U‘NWEESETY .
{waxing Lee

1973

LIBRARY

Michigan State
University

 

This is to certify that the
thesis entitled
Factors affecting liver fat accumulation and liver
hemorrhages associated with fatty liver-hemorrhagic

syndrome in laying chickens

presented by

Kwang Lee

has been accepted towards fulﬁllment
of the requirements for

Ph.D. degreein Poultry Science &
Institute of Nutrition

m 77%
/ /

 

 

Major p essor

Date May 1, 1973

0-7639

 

ABSTRACT

FACTORS AFFECTING LIVER FAT ACCUMULATION AND LIVER
HEMORRHAGES ASSOCIATED WITH FATTY LIVER-HEMORRHAGIC
SYNDROME IN LAYING CHICKENS

BY

Kwang Lee

Once a laying flock has been afflicted with the fatty
liver-hemorrhagic syndrome (FLHS), the economic loss, pri-
marily due to mortality resulting from liver hemorrhaging,
may be large, yet the exact cause(s) of this syndrome has/
have not been elucidated.

One of many difficulties in studying this problem has
been that the syndrome is very difficult to reproduce in
the laboratory at will since the exact causative agent(s)
is/are unknown.

The objectives of this study were to investigate fac-
tors affecting liver fat accumulation, to attempt to pro-
duce fatty liver-hemorrhagic syndrome, and to study the
relationship between liver fat infiltration and fatty
liver hemorrhaging.

A series of four experiments were conducted with a

commercial strain of Single Comb White Leghorn pullets.

Kwang Lee

When environmental temperature was "high" (27.80 C),
more livers appeared to be fatty, and incidence of liver
hemorrhages was greater than that obtained in "moderate"
(12.20 C) temperature.

Percent liver fat as well as total liver fat obtained
in the "high" environmental temperature was slightly higher
among birds fed a high carbohydrate diet as compared with
those fed a high fat diet. Statistical difference was
not obtained, however.

Restricted feeding followed by ag_libitum feeding
resulted in a reduction of both total liver fat and the
incidence of FLHS.

When a specific diet, Diet F, a commercial laying
formula, was given to birds, amounts of liver fat increased
markedly. Average feed consumption, final body weight
and hen-day egg production, however, were not statistically
different between birds fed Diet F and those fed the other
diet.

The highest incidence of liver hemorrhages and FLHS
resulted from one specific treatment where restricted
feeding was practiced with Diet F for four weeks in 30.60 C
environmental temperature then followed by an ad libitum
feeding in 22.20 C.

FLHS hemorrhages occurred in livers with high fat
content (high fat-liver) but high fat-livers were not al-

ways accompanied by severe hemorrhages. This showed that

Kwang Lee

high liver fat values may not necessarily be indicative
of FLHS.

FLHS livers were significantly heavier than livers that
did not have severe hemorrhages. Average feed consumption,
final body weight and hen-day egg production of birds having
FLHS hemorrhages did not differ from birds having livers
with a low fat content. Thus, it appears that changes in
feed consumption, body weight and egg production may not

be used as satisfactory indications of FLHS in laying birds.

FACTORS AFFECTING LIVER FAT ACCUMULATION AND LIVER
HEMORRHAGES ASSOCIATED WITH FATTY LIVER-HEMORRHAGIC

SYNDROME IN LAYING CHICKENS

BY

Kwang Lee

A THESIS

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

DOCTOR OF PHILOSOPHY

Department of Poultry Science
and

Institute of Nutrition

1973

as.
’b
5%?

is
\5\

The author wishes to express his sincere appreciation

ACKNOWLEDGEMENT

to Dr. Cal J. Flegal for his interest and guidance during
this period of study and research.

The author is grateful to Dr. J. H. Wolford for his
assistance in visual liver fat scoring and for his many
helpful suggestions.

The author also wishes to express his appreciation
to Drs. T. H. Coleman, H. C. Zindel of the Department of
Poultry Science and Dr. E. R. Miller of the Department of
Animal Husbandry for their guidance and careful review of
this manuscript.

Sincere thanks are also extended to Mr. Sulo Hulkonen
for photographic work.

To Dr. H. C. Zindel, the Chairman of the Poultry
Science Department, the author extends his appreciation
for the financial support provided in the form of a
Graduate Assistantship.

The author is particularly grateful to his parents
for their encouragement and moral support.

Finally, the author is indebted to his wife, Soon-Ja,
for her sacrifice, understanding and encouragement during
this period of study, and also for her contributions to this

research.

ii

TABLE OF CONTENTS
Page
ACKNOWLEDGEMENTS.......................... ii
LIST OF TABLES............................ v

LIST OF APPENDIX TABLESOOOOOOOOOCOOOOOOOOO Viii

I. INTRODUCTIONOOOOOOOOOOOOOOOOOOOOOOOOOOOOOO 1

II. REVIEW OF LITERATUREOOOOOOO0.0.0.0...0.... 3

III. GENERAL PROCEDURE.OOOOOOOOOOOOOOOOOOOOOOOO 13

IV. EXPERIMENTS............................... 15
A. Experiment I
1. Procedure......................... 15
2. Results and Discussion............ 21
B. Experiment II
1. Procedure......................... 28
2. Results and Discussion............ 29
C. Experiment III
1. Procedure......................... 38
2. Results and Discussion............ 43
D. Experiment IV
1. Procedure......................... 54

2. Results and Discussion............ 57

iii

Page

V. GENERAL DISCUSSION.OOOOOOOOOOOOOOOOOOOOOOOO 68

VI. SUMMARY...0.00000000000.............OOOOOOO 74

VII. LITERATURE CITED...O..O..OOOOOOO00.0.0.0... 77

VIII. APPENDIX....0.000000IOOOOOOOOOOOIOOIOOOOOOO 82

iv

LI ST OF TABLES

Table Page
1 Treatment description, experiments I and
II........O.....0.000..0.0.0.000...000...... 16
2 Percentage composition of diets used in

treatments 1' zand 3......0................ 18

3 Percentage composition of diets used in
treatments 4, S, 6 and 7.................... l9

4 Percentage composition of diets used in
treatments 8, 9, 10 and ll.................. 20

5 Visual liver fat SCOIG...................... 22

6 The effect of different sources of energy
on average liver fat score.................. 23

7 The effect of different sources of energy
on performance and some of the liver
characteristics of laying birds............. 25

8 Analyses of variance for initial body weight,
final body weight and the percentage of
liver fat in experiment I................... 26

9 The effect of different sources of energy
on the performance of laying birds kept in
a "high" environmental temperature
(27.80 C)................................... 30

10 Analyses of variance for feed consumption
and hen-day egg production in experiment

110.00.000.00...0.000000000000000000000000.0 31

ll Analyses of variance for initial and final
body weights in experiment II............... 32

12 The effect of different sources of energy
on some of the liver characteristics of
laying birds kept in a "high" environmental
temperature (27.80 C)....................... 34

13 Analyses of variance for several liver
characteristics in experiment II............ 35

V

Table Page

14 The effect of a high carbohydrate and a
high fat diet on average feed consumption,
final body weight, egg production and
liver fat accumulation...................... 36

15 Treatment description, experiment III....... 40

16 Amount of feed allowed per day during
restricted feeding period................... 41

17 comPOSition Of Diet F..........0.0.......... 42

18 The effect of feed restriction on average
body weight and hen-day egg production...... 44

19 The effect of restricted feeding followed
by ad libitum feeding on the performance
Of EYlng 61rds....OOOOOOOOOOOOIOOOOOOOOOOOO 46

20 Analyses of variance for the performance of
laying birds in experiment III.............. 47

21 The effect of two diets, and a restricted
feeding followed by ad libitum feeding
on several liver characterist1cs............ 49

22 Analyses of variance for several liver
characteristics in experiment III........... 50

23 Comparison of average feed consumption and
liver fat content in relation to presence
or absence of FLHS hemorrhages.............. 53

24 Treatment description, experiment IV........ 56

25 The effects of feed restriction in dif—
ferent environmental temperatures on average
body weight and egg production.............. 58

26 Analyses of variance for the performance
of laying birds during the feed restric-
tion period in experiment IV................ 59

27 The effect of restricted feeding followed
by ad libitum feeding in different environ-
mental temperatures on the performance of
laying birds................................ 60

28 Analyses of variance for the performance

of birds during the ag_libitum feeding
period in experiment IV..................... 61

vi

Table Page

29 The effect of two environmental tempera-
tures and restricted feeding followed by
ad libitum feeding on several liver
characteristics............................. 63

30 Analyses of variance for several liver
characteristics in experiment IV............ 64

31 Comparison of liver fat content and the per-

formance of birds in relation to presence
or absence of FLHS hemorrhages.............. 66

vii

LIST OF APPENDIX TABLES
Table Page

1 Composition of vitamin-trace mineral
premixture for laying ration (Diets l
to ll)...0......OOOOOOOOOOOCCOOOOOO0.00.... 82

2 Composition of pullet developer diet....... 83

3 Correlation coefficients of the variables
obtained during the final dd libitum
feeding period (28 to 40 weeks of age) of
experiment III............................. 84

4 Composition of Z-4 laying formula.......... 85
5 Correlation coefficients of the variables
obtained during the ad libitum feeding

period (36 to 40 weeFE of agel of experi-
ment IVOOOCC.O.......0....0...............O 86

viii

I. INTRODUCTION

Fatty liver syndrome in mammals has been studied since
1932.

In poultry, however, fatty liver is a fairly new
disease problem recognized during the develOpment of a
new management structure, viz., high bird-density cage sys-
tem.

Several differences in fat metabolism between rats
and chickens have been recognized. Rats synthesize most
of their fatty acids in the adipose tissue. Chickens,
however, synthesize most of their fatty acids in the liver.
Causes of fatty liver in rats and chickens may not be the
same. A lack of choline chloride in the diet results in
fatty liver in rats but not necessarily in chickens.

The significance of liver hemorrhages has not been
discussed until recently. It has been shown that mortal-
ity can result from severe liver hemorrhaging but not
necessarily from the fatty liver ESE g3.

Neither the exact causes of fatty liver (high fat-
liver) nor liver hemorrhages in chickens have been elu—
cidated.

One of the difficulties in studying fatty liver-

hemorrhagic syndrome is the difficulty in reproducing

it in the laboratory, at will; accordingly, the objectives

of this study were: (1) to investigate nutritional and
environmental factors affecting liver fat accumulation;

(2) to attempt to produce fatty liver-hemorrhagic syndrome
in the laboratory and; (3) to study the relationship between

liver fat infiltration and fatty liver hemorrhaging.

II. REVIEW OF LITERATURE

One of the classical examples of fatty liver syndrome
can be found in choline deficiency in rats. As early as
1932, Best EE.E£° (1932a) observed an accumulation of fat
in hepatic cells of rats which were raised on a diet con-
taining 2.5 grams of fairly saturated fat. This condition
was cured by an oral administration of choline chloride
(Best EE.El-I 1932b). Chalvardjian (1969) noticed an in-
crease in hepatic triglyceride pool in livers of rats fed
a choline deficient diet. The C-l6 (saturated and monoenoic)
fatty acids were major components of triglyceride in liver
lipids of rats fed a fat-free choline deficient diet. How-
ever, when fat was added to the diet, the fatty acid pattern
of hepatic lipids was similar to that of dietary fat
(Chalvardjian, 1966).

The lipotropic mechanism of choline is complicated.

Due to the fact that the addition of choline resulted in

an increase in body weight and deposition of fat in the

body of rats, Maclean 2E.2l° (1937) suggested that the
presence of choline in the diet favored the normal distri-
bution of fat between the liver and body depots. Chalvardjian
(1969) noticed a decrease in transport of triglycerides

and phospholipids from the liver to serum in rats fed a

choline deficient diet. Perlman and Chaikoff (1939) re-
ported that choline accelerated the rate of formation of,
as well as a rapid removal of, phospholipids in the liver.

The lipotropic effect of methionine was reported by
Tucker and Eckstein (1937). When methionine was added to the
choline deficient diet, fatty liver incidence in rats was
significantly reduced. The finding that cysteine and homo-
cystein (sulfur containing amino acids) did not exert their
effect in reducing fat content in the liver led them to the
conclusion that the lipotropic action of methionine was due
to the presence of a labile methyl group which was not pres-
ent in the other two amino acids.

The metabolic interrelationship of methionine and choline
in the rat was reported by Du Vigneaud EE.El° (1940). They
demonstrated the biological transfer of the intact methyl
group from methionine into the choline molecule (Transmethyl-
ation reaction). In the following year, they stated that
the lipotrOpic action of methionine in a choline deficient
situation might be due to the presence of the labile methyl
group of methionine which was used for the synthesis of
choline (Du Vigneaud EE.§l°I 1941). Similar observations
were made in chickens. Leach 2E.3lf (1966) reported that
when there were sufficient methyl groups in the diet, hens
were able to synthesize choline. However, the addition of
high levels of choline to the diet was not effective in

preventing fatty liver in chickens (Fisher and Weiss, 1956).

Hegsted SE El' (1941) reported that choline deficient
chicks did not show fatty livers; however, choline was needed
to prevent perosis.

Chickens have been recognized to be different from
rats in several aspects of lipid metabolism. O'Hea and
Leveille (1968) found that rats' adipose tissue was much
more active in synthesizing fatty acids than chicks' adipose
tissue. They also found that the incorporation of glucose-

U-Cl4

into fatty acids was stimulated by insulin in rat
adipose tissue but not in chick adipose tissue. Leveille
(1967) reported that adipose tissue was the major site of
fatty acid synthesis of rats. But due to the fact that the

14 and acetate-l-C14

incorporation rates of both glucose-U-C
into fatty acids were much higher in the liver than in the
adipose tissue of chicks, Leveille EE.El' (1968) concluded
that the liver was the major site of fatty acid synthesis
in chicks.

The age and sex of chickens have been recognized to
influence lipid metabolism in the liver. Leveille (1969)

found that acetate-l-Cl4

incorporation into fatty acids

was greater in the liver of chicks than that of hens. How-
ever, no statistical significance was noticed. Saloma
EE.2$° (1969) reported that the amount of linoleic acid in
triglyceride was significantly higher in the liver of laying

hens as compared to that of non-layers. They also reported

that the major fatty material accumulated in the liver of

laying hens was triglyceride. Lorenz EE.2£° (1937-1938)
observed a marked increase in blood lipid concentrations,
eSpecially triglyceride, in actively laying hens as compared
to that of immature female birds. Blood lipid concentrations
of mature male, of immature male and of immature female
chickens were not different from each other. These find-
ings indicated that the rise in lipid level in the blood was
related to the ovarian activity. However, once the female
bird attained sexual maturity, increased blood lipid level
was not related to the intensity of egg production. Shortly
after, Lorenz §E_§l, (1938) observed a marked increase in
neutral fat in the liver at the onset of maturity of the
female chicken. But phospholipid and cholesterol ester
content were not changed.

O'Hea and Leveille (1969), when studying the origin of
plasma fatty acids, found these originated from the liver

14

since the pattern of distribution of glucose-U-C and

14 in plasma and in liver was similar after

acetate-l-C
injection of these materials. Over 75 percent of the
plasma lipid radioactivity was found in triglyceride.
They also reported that the main transport form of lipid
from liver to adipose tissue was 8 or low-density lipo-
protein since these components contained over 66 percent
of plasma radioactive lipid. Duke EE.E£° (1968) observed

no significant difference in plasma protein levels between

two dietary groups of birds (the non-control group developed

fatty liver syndrome) which led them to conclude that plasma
protein level was not a satisfactory index of developing
fatty liver syndrome.

Oser (1965) pointed out the existence of different
types of fatty liver in animals since the central vein area
of the liver lobule was the first place fat appeared in
choline deficiency, whereas periportal was the first place
in starvation and in low-protein feeding. According to
WOhl and Goodheart (1970), Peters and Van Slyke in 1946
classified fatty livers as physiologic and pathologic type.
The physiologic type of fatty liver resulted from the mobi—
lization of large amounts of fat from depots to liver to meet
unusual requirements for lipid oxidation. This type of
fatty liver occurred in fasting, carbohydrate starvation
or in pancreatic diabetes. The pathologic type of fatty
liver resulted from the impaired turnover of lipid in the
liver by poisoning (chloroform, carbon tetrachloride, white
phosphorous or a massive dose of alcohol) or by a dietary
disorder (choline deficiency, amino acid imbalance or
chronic alcoholism).

"Fatty liver syndrome" in chickens was first reported
by Couch (1956) and was characterized by 1) increased body
weight, 2) decreased egg production, 3) excessive amount
of abdominal fat, 4) fatty livers, 5) capillary hemorrhages
in the liver and hematoma. Reedy (1968) also characterized

birds afflicted with fatty liver syndrome as: "very

healthy appearing birds suffering a gradual decrease in
production," having a heavy body weight, having an exces-
sive amount of abdominal fat, having an enlarged, fatty and
friable liver and being sensitive to stress factors. Parker
and Deacon (1968) stated that fatty liver syndrome occurred
in all breeds of commercial hens fed all kinds of commer-
cial diets.

Price gE_gl. (1957) and Barton 25.3l. (1966) observed
a higher incidence of fatty liver problem in the caged
layers than floor layers fed the same ration; however,
bird density in a cage did not affect the total amount
of lipid in the liver (Owings SE El-v 1967).

According to Couch (1964), Delaplane was unable to
transmit fatty liver condition through feeding fecal materi-
als, transplanting respiratory discharges and injecting
blood of birds having fatty liver syndrome into normal
birds.

Hamilton and Garlich (1970) reported that fatty liver
syndrome could be produced by feeding aflatoxin. Later,
Hamilton 22.2l' (1972) demonstrated that feeding 1.0 ppm
of aflatoxin to turkey poults resulted in a higher total
lipid/liver as compared to the control group (0 ppm). How-
ever, the liver size was smaller in the aflatoxin fed group
than the control fed group.

Dudley 23 El: (1961) reported that as the dietary

cholesterol level was increased, liver cholesterol

content, total liver lipid and liver size also increased
in young male chicks.

Turk 33,31. (1958) observed an increase in the amount
of liver fat as the energy level of the diet was increased.
Barton EE.El3 (1966) made a similar observation that birds
fed a high energy ration developed fatty liver syndrome
which was prevented by feeding a low energy-high fiber
diet. Morrison 25.2l' (1970) also reported that a signifi-
cantly higher content of liver fat was obtained in birds
fed a diet containing 3080 Kcal. M.E./kg. diet in comparison
to a group fed a 2640 Kcal. M.E./kg. diet.

Quisenberry 2E.il° (1967) reported that even though
body weight was increased as a result of feeding a high
energy diet, fatty liver syndrome neither resulted nor
was prevented by dietary calorie level. Donovan and
Balloun (1955) also reported that energy level in the diet
did not affect the amount of fat in the liver of chicks.

In addition, they pointed out that the level of dietary
protein and amount of lipid content in the liver of chicks
was inversely related.

Howes and Fitzgerald (1966) found that as the dietary
fat level was increased, liver fat also increased in coturnix
quail. Weiss and Fisher (1957) also found an increased
deposition of abdominal fat, as well as liver fat, when birds
were fed a diet with 5 percent added fat in comparison to

a no added fat group. On the contrary, Treat EE.E£- (1960)

10

noticed the highest total liver weight and the percent liver
fat from birds fed a diet containing no added fat in com-
parison to groups fed diets which contained 2.5 to 5 percent
added fat. Due to the fact that the higher amount of fat
was deposited in the livers of rats receiving a fat free
diet, Harper 22.2l' (1954) concluded that dietary fat ESE
g3 did not accumulate to any greater extent in the liver

and liver fat arose from the conversion of carbohydrate or
protein to fat. McDaniel 22.2l° (1959) also reported that
added fat as an increased source of energy did not affect
liver weight or liver fat of laying hens.

Nesheim 23.3l' (1969) studied extensively fatty liver
problems in laying hens. Some of the points of their re-
port were:

1) Fatty liver mortality occurred almost entirely
during the months of April, May and June which
suggested that faulty adjustment of energy con-
sumption during this period may be involved in
causing this problem.

2) The best way to produce fatty livers might be
to force feed diets in an amount above that nor-
mally consumed dd libitum.

3) High levels of liver fat EE£.§E.maY not be the prob-
lem in a laying flock as many hens that had high
levels of liver fat were good performers and

furthermore, hens did not die because of high

11

levels of liver fat but they died due to a
massive liver hemorrhage.

In the following year, Nesheim and Ivy (1970) again
stated that fatty liver might be developed due to an ex-
cess consumption of energy which might be the result of an
inability of some hens to regulate energy intake. They
also suggested to call fatty liver syndrome as "liver
hemorrhage syndrome."

The term "fatty liver-hemorrhagic syndrome (FLHS)"
was used by Wolford and Polin (1972a) who produced FLHS
in laying hens by a force-feeding method. Groups of birds
were force-fed feed in an amount of 50, 100, 112.5, 125 and
150 percent of control ad libitum fed group. Energy and
protein content of the diet were 3095 Kcal. M.E./kg. and
17.5 percent, respectively. Within three weeks, birds that
received the increased amount of feed showed an increased
incidence and severity of hemorrhage in the liver. They
concluded that overconsumption of feed (energy) was one of
the probable causes of FLHS in laying hens.

Couch (1956) recommended a premix for the treatment of
fatty liver syndrome which contained 5,000 to 10,000 10
of vitamin E, a minimum of 500 grams of choline chloride
and 12 milligrams of vitamin B12 per ton of feed. The
effect of this type of premix was not always satisfactory
for preventing or alleviating fatty liver syndrome as re-

ported by Barton (1967), Quisenberry 22.2l- (1967), Deacon

12

(1968) and Parker and Deacon (1968). However, Bull (1968)
noticed a more than 50 percent reduction of liver fat
among flocks which had a typical fatty liver syndrome when
vitamin 812 and vitamin E were added to the diet along
with inositol. Reed EE.El° (1968) also found that adding
inositol (% lb./ton) to the diet decreased percentage of
liver fat. Further decrease in percentage of liver fat
was noticed when the combination of inositol (2.5 lbs./ton),
choline, vitamin E and vitamin 312 was added to the diet.
Nelson (1968) reported that the addition of inositol
(2 lbs./ton) caused a significant decrease in percentage
of liver fat and a significant increase in liver moisture
content of broiler breeder hens. On the other hand, Ragland
EEHEl' (1970) reported that addition of inositol (1,000
ppm) did not affect feed intake, egg production, body weight
and percentage of liver fat of laying hens.
Jensen 23 El: (1970) introduced selenium as a new
lipotropic agent. They observed a significant reduction
in liver weight and total liver lipid when selenium was
added to a diet of laying hens. On the contrary, Griffith
and Schexnailder (1972) reported that addition of selenium,
vitamin E or inositol to the laying ration did not signifi-
cantly reduce liver fat while addition of choline and vita-

min B12 was very effective.

III. GENERAL PROCEDURE

With the exception of part of experiment IV (treat-
ments 2 and 4), all experiments were conducted in house
8-B (windowless) at the Michigan State University Poultry
Science Research and Teaching Center, where two gas hovers
were provided for the purpose of elevating environmental
temperature whenever needed. A thermostat, an exhaust fan
and an automatic light controlling device were also pro-
vided in the house.

A commercial strain of Single Comb White Leghorn
(S.C.W.L.) pullets (De Kalb 131) were placed in triple deck
laying cages (20.3 cm x 40.6 cm).

Body weights were measured at the beginning and at
the end of each experiment. During all trials water was
given ad libitum. Dead birds were sent to the Michigan
State University diagnostic laboratory to determine the
cause of death.

Birds were sacrificed by cervical dislocation, and
their livers were collected for fat analysis. Liver sam-
ples were stored in a freezer (-150 C) until analyzed.
Liver fat analysis was done according to the method de-

scribed by Folch 32 El. (1957).

13

14

Those livers that contained 15 percent or more fat
(wet basis) were considered as high fat-livers since the
lowest percent liver fat with FLHS hemorrhages was found
to be 15 percent, which was equivalent to 7.05 grams of
fat per liver. When a liver contained 15 percent or more
fat and also had more than 15 hemorrhages and/or hematoma
(Wolford and Polin, 1972a), that liver was classified as
FLHS liver in these experiments.

Data for percent hen-day egg production and percent
liver fat were transformed to arcsin prior to statistical
analysis. Analyses of variance and correlation coefficients
were computed at the computer center of Michigan State
University. Dunnett's "t" test (Kirk, 1968) was used
whenever applicable for comparisons between the control

and treatments.

IV . EXPERIMENTS

A. Experiment I

1. Procedure:

 

The primary purpose of conducting this experiment
was to study the effect of different sources of energy
(carbohydrate and fat) on the accumulation of fat in the
liver of laying birds.

One hundred thirty—two, 22-week old S.C.W.L. pullets
were randomly assigned to each of three replications in
Leach treatment and were held in a thermocontrolled house
(12.20 C). Electric lights in the house were automatically
turned on and off at 6:00 A.M. and 8:00 P.M., respectively,
to provide the birds with a minimum of 14 hours of light
daily. During the experimental period of 12 weeks (22 to
34 weeks of age), two pullets were kept in each of 66
cages and were fed diets dd libitum.

Based on different sources of carbohydrate or fat as
a primary energy supplement, ll dietary treatments were
used (Table l).

Diets used in treatments 1, 2 and 3 (Control) were
corn-soy type formulae. Dietary levels of energy and/or
protein of these three treatments were different from one

another. Both the energy and protein level of the diet

15

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in treatment 2 were higher than those of treatment 1, yet
calorie-protein ratios were calculated to be identical.
Treatment 3 was different from treatment 2 with respect
to a lower dietary protein. Dietary energy levels of these
two treatments were the same, however.

Diets having corn, wheat, milo, barley or cats as a
primary energy source were designated to be treatments
3, 4, 5, 6 and 7, respectively. Corn oil, safflower oil,
herring oil or lard were used as fat sources of energy,
and corresponding diets were labelled as treatments 8, 9,
10 and 11, respectively.

Diets 3 to 11 were calculated to be isocaloric (3130
M.E. Kcal./kg.) and isonitrogenous (15 percent protein)
(Table 1).

Composition of diets can be seen in Tables 2, 3 and

Due to the fact that diets 6 to 11 were formulated
to contain fairly high levels of fat, mixing feed for those
diets was completed by two separate operations. First,
all the dietary ingredients including 5 percent added
fat were mixed in a feed mixer and then the feed was re-
moved from the mixer and was placed on a concrete floor.
In the second step, the remaining fat was added to the
previously mixed feed, mixed with a shovel and then feed
was forced through a wire mesh (0.7 cm x 0.7 cm). This

hand-mixing was done at least three times per diet to

18

Table 2. Percentage composition of diets used in treatments

 

 

l, 2 and 3
Ingredients Diet 1 Diet 2 Diet 3
Ground yellow corn 71.00 63.06 66.10
Soybean meal, 49% protein 15.00 18.71 15.90
Alfalfa meal, 17% protein 1.00 1.00 1.00
Distillers dried solubles, corn 1.00 1.00 1.00
Fishmeal, menhaden 1.00 1.00 1.00
Dried whey 0.50 0.50 0.50
Wheat bran 2.00 2.00 2.00
Ground limestone 6.04 5.80 5.91
Dicalcium phosphate 1.60 1.50 1.50
Corn oil 4.56 4.23
Salt, iodized 0.20 0.20 0.20
Vit.-min. premix* 0.58 0.58 0.58
Methionine 0.08 0.08 0.08
Total 100.00 100.00 100.00

 

*Refer to Appendix Table l.

l9

 

 

Table 3. Percentage composition of diets used in treatments
4, 5, 6 and 7
Ingredients Diet 4 Diet 5 Diet 6 Diet 7
Wheat 67.30
Milo 67.36
Barley 62.58
Oats 62.58
Soybean meal, 49% protein 9.82 12.56 10.98 10.98
Alfalfa meal, 17% protein 2.00 2.00 2.00 2.00
Distillers dried solubles,
corn 2.00 2.00 2.00 2.00
Fishmeal, menhaden 2.00 2.00 2.00 2.00
Dried whey 0.10 0.10 0.10 0.10
Wheat bran 0.10 0.10 0.10 0.10
Ground limestone 6.28 6.20 5.91 5.97
Dicalcium phosphate 1.90 1.90 1.50 1.50
Corn oil 7.64 4.92 11.97 11.91
Salt, iodized 0.20 0.20 0.20 0.20
Vit.-min. premix* 0.58 0.58 0.58 0.58
Methionine 0.08 0.08 0.08 0.08
Total 100.00 100.00 100.00 100.00

 

*Refer to Appendix Table l.

20

 

 

Table 4. Percentage composition of diets used in treatments
8, 9, 10 and 11
Ingredients Diet 8 Diet 9 Diet 10 Diet 11

Ground yellow corn 30.00 30.00 30.00 30.00
Soybean meal, 49% protein 21.10 21.10 21.10 21.10
Alfalfa meal, 17% protein 2.00 2.00 2.00 2.00
Distillers dried solubles,

corn 3.00 3.00 3.00 3.00
Fishmeal, menhaden 0.50 0.50 0.50 0.50
Dried whey 2.50 2.50 2.50 2.50
Wheat bran 3.00 3.00 3.00 3.00
Ground limestone 5.90 5.90 5.90 5.90
Dicalcium phosphate 1.50 1.50 1.50 1.50
Corn oil 15.89

Safflower oil 15.89

Herring oil 17.39

Lard 16.50
Salt, iodized 0.20 0.20 0.20 0.20
Vit.-min. premix* 0.58 0.58 0.58 0.58
Methionine 0.08 0.08 0.08 0.08
Solkafloc 13.75 13.75 12.25 13.14

Total 100.00 100.00 100.00 100.00

 

*Refer to Appendix Table l.

21

make sure that the feed ingredients were properly mixed
with fat.

Feed consumption and body weight were measured bi-
weekly and egg production was recorded daily.

In order to examine the degree of fat accumulation
in the liver, one bird from each replication in each treat-
ment was sacrificed at the end of both the sixth (28 weeks
of age) and ninth (31 weeks of age) weeks of the experi-
ment and were subjected to visual liver fat scoring. The
scoring system is shown in Table 5.

At the end of the twelfth week of the experiment (34
weeks of age), all remaining birds were sacrificed and

livers were collected for fat analysis.

2. Results and Discussion:

 

The effect of different sources of energy on the
average visual liver fat score is presented in Table 6.

The data revealed that the average visual liver fat
scores of the livers in all treatments were not high enough
to indicate those livers as having high fat content. Livers
with high fat content (high fat-liver) may be indicated
by a liver score of 3.0 or above (Table 5). Also, there
was no strong indication of further deposition of fat in
the liver when liver samples were collected in two dif-
ferent periods.

None of the livers collected at the end of the sixth

week (28 weeks of age) of the experiment were scored as

22

Table 5. Visual liver fat score*

 

 

 

Liver
Score Texture Color
0.5 and 1.0 Firm Mahogany
1.5 and 2.0 Firm Slight yellow in mahogany
2.5 and 3.0 Less firm Yellow, little mahogany

3.5 and 4.0 Ruptured easily Very yellow, no mahogany

 

*WOlford and Polin (1972b).

23

Table 6. The effect of different sources of energy on
average liver fat score

 

Avg. visual liver fat score

 

Treatment Ingredient 28 wks.* 31 wks.*
1 Corn 1.3 1.2
2 Corn 1.2 0.8
3 (Control) Corn 1.2 1.7
4 Wheat 1.2 1.2
5 Milo 1.0 2.0
6 Barley 0.8 1.3
7 Oats 1.2 1.4
8 Corn oil 1.0 1.2
9 Safflower oil 1.3 1.0
10 Herring oil 1.2 1.3
11 Lard 1.0 1.3

 

*Age of birds.

24

3.0 or above, and only two livers were scored as 3.0 at
the end of the ninth week (31 weeks of age).

The effect of different sources of energy on the per-
formance and some of the liver characteristics are shown
in Tables 7 and 8.

The largest average feed consumption and the heaviest
average body weight resulted from the group of birds fed
the corn-soy diet of treatment 1. The highest average
egg production, however, was recorded from the birds fed
the barley diet.

When birds were fed the diet that contained herring
oil, the smallest average feed consumption and the lowest
average egg production resulted.

It has been reported that birds eat to satisfy their
energy requirement. Indirect evidence of this is that
an increase in dietary energy from 2600 to 3350 calories
(M.E.) per kilogram of diet will decrease feed consumption
from 117 to 90 grams per hen per day during a 20 week laying
period from 22 to 42 weeks of age (Scott 2E.El°' 1969).

In this experiment where isocaloric-isonitrogenous
diets (treatments 3 to 11) were used, average feed
consumption of birds in different dietary groups was dif—
ferent (56.3 to 107.1 gm./bird/day). This was partic-
ularly true when herring oil was included in the diet.
This result implies that there are some factor(s) in the feed
ingredients other than dietary energy which may influence

feed consumption.

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25

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27

Livers obtained on the last day of the experiment
revealed that four livers (from treatments 1, 4, 5 and
10) contained 15 percent or more fat (wet basis).

Visual liver fat scoring system was also utilized
on the terminal day of the experiment. A highly signifi-
cant correlation (r = +0.74) between the visual liver fat
score and the chemically determined percent liver fat was
noted.

Average data for liver characteristics indicated that
none of the diets produced high fat-liver or fatty liver-
hemorrhagic syndrome (FLHS) equivalent to those observed
in field cases. Some incidence of liver hemorrhages, how-
ever, was observed among birds fed diets containing corn
(control), wheat and herring oil.

In this experiment 3.8 percent mortality was recorded

but none of the birds died of FLHS.

B. Experiment II

1. Procedure:

 

Since most of the fatty liver mortality in field cases
have been observed during spring-summer months as reported
by Nesheim 2E.2l- (1969), a "high" environmental tempera-
ture (27.80 C) was employed in this experiment to see
whether a "high" temperature along with different sources
of energy affected liver fat accumulation.

Experiment II was a duplicate of the first experiment
with the following exceptions: A high environmental
temperature (27.80 C), older birds (29 weeks of age at the
beginning of the experiment), less frequent intervals for
measuring body weight and feed consumption (4 week inter-
val) and no sacrifice of birds until the termination of
the experiment.

Dietary treatments and the composition of diets used
in this experiment were identical to those of the previous
experiment (Tables 1, 2, 3 and 4).

Since the design of this experiment was split—plot,
statistical analysis of the data for feed consumption and
egg production was computed by analysis of variance for
split-plot design (Kirk, 1968). Scheffe's "s" test (Kirk,
1968) was utilized to compare non-paired means (i.e.: high

carbohydrate diet vs. high fat diet).

28

29

2. Results and Discussion:

 

The effect of the different sources of energy on the
performance of laying birds kept in a "high" environmental
temperature may be seen in Tables 9, 10 and 11.

The results showed that those birds fed the diet that
contained wheat consumed the most feed and were the largest
in average body weight. The highest average egg produc-
tion, however, was obtained from the birds fed the diet
that contained milo. As in the case of the previous ex-
periment, birds fed the diet that contained herring oil
consumed the least amount of feed and laid the least number
of eggs.

The results also indicated that as long as dietary
levels of energy and protein remained the same, the corn
diet (control) could be replaced by all other grain diets
without adversely affecting laying performance.

With the exception of the group of birds fed the
diet that contained lard, all birds fed diets that con-
tained fat as the primary energy source consumed signifi-
cantly less feed and produced fewer eggs than the control
group.

Liver samples collected on the terminal day of the
experiment revealed that 16 livers (four from treatment 1,
one from treatment 2, three from treatment 3, three from
treatment 4, two from treatment 5, one from treatment 10

and two from treatment 11) contained 15 percent or more

30

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31

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HH ucmﬁmuomxm cm coHu
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32

Table 11. Analyses of variance for initial and final body
weights in experiment II

 

 

 

Source of Degrees of
variation freedom Mean squire

Initial body wt. Final body wt.

29 wks. 41 wks.

Total 125
Treatment 10 21720.7 110887.l*
Replication 2 47403.2 203306.2*
T x R 20 30420.5 44713.2
Error 93 23922.0 50837.0

 

*Significant at P < 0.05.

33

fat (wet basis) and four livers (two from treatment 1 and
two from treatment 4) had hematomas.

The effect of different sources of energy on some of
the liver characteristics of laying birds kept in a "high"
environmental temperature can be seen in Tables 12 and 13.

The results showed that livers obtained from all of
the treatments with the exception of treatment 1 were signi-
ficantly lighter in average wet weight than those of the
control group. Average percent liver fat and total liver
fat values were also smaller than those of the control
in most cases.

Eighteen percent of the birds that received the corn
diet (control) and 9 percent of the birds that received
the wheat diet were found to have FLHS hemorrhages. Con-
sidering the fact that average percentages of liver fat
for those treatments (as well as in other treatments) were
not very high, the occurrence of FLHS might not be due to
those diets.

In this experiment, treatments 3, 4, 5, 6 and 7, and
treatments 8, 9, 10 and 11 were collectively referred to
as a high carbohydrate and a high fat diet, respectively.
The effect of a high carbohydrate and a high fat diet on
average feed consumption, body weight, egg production and
liver fat accumulation can be seen in Table 14.

Under the conditions of this experiment, birds fed a
high carbohydrate diet consumed significantly more feed

and produced more eggs than the birds fed a high fat diet.

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34

 

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35

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36

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37

The average values for final body weight and liver
fat (percent and total) were also slightly higher for those
birds fed a high carbohydrate diet as opposed to a high
fat diet. Statistical difference due to diets was not
obtained, however.

In this experiment 4.5 percent mortality was recorded.

However, mortality due to FLHS was nil.

C. Experiment III

1. Procedure:

 

The purpose of conducting this experiment was to try
to produce fatty liver-hemorrhagic syndrome in laying
birds by taking advantage of the results of the previous
experiment that showed a high carbohydrate diet in a "high"
environmental temperature seemed to favor accumulation of
liver fat. A restricted-3d libitum feeding system was em-
ployed to induce overeating during the Ed libitum period,
since WOlford and Polin (1972a) observed FLHS among force-
fed hens which received an excess amount of feed.

This experiment was conducted with 60 S.C.W.L. pullets
which were randomly assigned into three replications of five
treatments. During the experimental period of 26 weeks,
all birds were kept in individual cages.

The experimental period was divided into the following
three phases:

First phase (phase 1): 14-20 weeks of age

Second phase (phase 2): 20-28 weeks of age

Third phase (phase 3): 28-40 weeks of age

Environmental temperatures during phases 1 and 2 were
maintained at 15.90 C and during phase 3 at 30.60 C (actual

average temperatures).

38

39

A 9-hour artificial light day (7:30 A.M. to 4:30 P.M.)
was provided from 14 to 20 weeks of age; thereafter, 14
hours of light (6:00 A.M. to 8:00 P.M.) were provided daily.

During the first phase, birds in all treatments were
fed a pullet developer diet (composition of pullet developer
diet is listed in Appendix Table 2).

From 20 to 40 weeks of age (phases 2 and 3) birds in
all treatments (with the exception of those in treatment 2)
received the diet used in treatment 3 (control) of previous
experiments. The diet used in treatment 2 was a commercial
laying formula. (This ration is presently not available.)
According to Barton (1967), birds that received this ration
had experienced some problems with fatty liver in the field.
For convenience, this diet will be referred to as Diet F.
Diet F was made and was included in this experiment to deter-
mine if dietary factor(s) other than feed consumption (energy
intake) is/are involved in producing FLHS.

The birds in treatments 1 and 2 received ad libitum
feeding throughout the experiment, while a reduced amount
of feed was given each day to the birds in treatments 3
and 4 during the first and second phases, respectively.
The birds in treatment 5 received a reduced amount of feed
daily during both phases 1 and 2, and then were allowed to
eat ad libitum during the third phase.

Treatment description, the amount of feed allowed per
day during the restricted feeding period, and the composition

of Diet F can be seen in Tables 15, 16 and 17, respectively.

40

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.mxz osumm .mxz mmuom .mxa omuvH
momma mo mma
HHH ucmEHHomxm .coHumHuommc ucoEummuB .mH anme

41

Table 16. Amount of feed allowed per day
during restricted feeding period

 

 

 

 

Age of bird Feed restriction
weeks Pounds feed/100 pullets/day
14-16 13
16-18 14
18-19 15
19-20 16

20-28 19

 

42

Table 17. Composition of Diet F

 

 

 

Ingredient Percent

Ground yellow corn 75.00
Soybean meal, 49% protein 11.90
Alfalfa meal, 17% protein 0.90
Meat and bone meal, 50% protein 2.20
Fishmeal, menhaden 0.40
Dried fish soluble 0.10
Feather meal 0.20
Blood meal 1.00
Ground limestone 5.85
Dicalcium phosphate 1.40
Salt, iodized 0.35
Animal tallow 0.50
Vit.-min. premix* 0.20
Total 100.00

Calculated analysis
Kcal. M.E./kg. diet 3004

Crude protein, % 15.2

 

*Supplied the following per kg of diet: Vitamin A,
3520 IU; Vitamin D3, 1100 I.C.U.; Riboflavin, 3.1 mg;
Pantothenic acid, 5.3 mg; Niacin, 11.0 mg; Choline
chloride, 171.6 mg; Folic acid, 0.44 mg; Vitamin B1 ,
5.28 ug; Vitamin E, 2.2 IU; Menadione sodium bisulfite,
0.7 mg; Manganese, 25.7 mg; Iodine, 0.40 mg; Copper,
0.16 mg; Cobalt, 102 ug; Zinc, 20 mg; Iron, 10.0 mg;
Magnesium, 200 mg.

 

 

 

43

Body weight and feed consumption were measured at the
end of 17 and 20 weeks of age for phase 1, 24 and 28 weeks
for phase 2 and 34 and 40 weeks for phase 3. Egg produc-
tion was recorded daily and egg weight was measured on
three consecutive days of every week (1:30 P.M. on Tuesday,
Wednesday and Thursday) during 20 to 40 weeks of age. At
the end of 40 weeks of age, birds in all treatments were
sacrificed for liver fat analysis.

The data for feed consumption and body weight obtained
during the first and second phases were not subjected to
the statistical analyses since a fixed amount of feed was
given to birds during those periods.

Student's "t" test was used to compare FLHS livers and
high fat-livers without having severe hemorrhages, and be-

tween FLHS livers and low fat-livers.

2. Results and Discussion:

 

The effect of feed restriction on average body weight
and hen-day egg production is shown in Table 18.

As expected, the most obvious effect of feed restric-
tion was a reduction of body weight during the feed restric-
tion period (treatment 3 in phase 1 and treatment 4 in
phase 2). During phase 2, birds that received a restricted
amount of feed laid 4 percent less eggs than the control
birds, which received dd libitum feeding.

When feed restriction was terminated, birds consumed

7 more grams of feed, on the average, per day and laid 4

44

.omm mo mxom3 somuusom

 

 

 

 

 

N
.moqu mo mmcH
m.m> mhmH mmNH NNOH N.mm m.mw m
N.H> mNmH mmvH NNOH H.Hm ¢.mm v
«.mh HmmH vaH mmoH m.¢NH ¢.mm m
v.mn NmmH ome ¢m0H m.mNH H.¢m N
«.mn ommH mmvH mm0H o.NHH m.mm AHouucouv H
:3 ES 2:3 A89 2:8 2:3
. useﬁummue
.mxz mNION .mx3 mN .mxz ON HwHuHcH .mxz mNION .mx3 ONieH
.ponm mmo Monica: .m>¢ uano3VNoon .m>4 Mmc\cuHmewmm mm><
GOHH

loopoum mmm moored: can uanms moon mmmum>m co COHHOHHumoH comm mo uommmm one .mH mHnma

45

percent more eggs than did the control birds during the
next eight weeks of the ad libitum feeding period (treat-
ments 3 vs. 1 in phase 2).

Birds fed Diet F consumed 6 more grams of feed (on
the average) daily and produced 4 percent more eggs than
did the birds in the control group during phase 2. This
increased feed consumption might be due to lower energy
content of Diet F in comparison to the control diet.

At the end of phase 2, birds in all treatments were
allowed to eat dd_1ibitum in the elevated environmental
temperature (30.60 C) for a period of 12 weeks.

The effect of restricted feeding followed by 29.11237
Egg feeding on the performance of laying birds is shown
in Tables 19 and 20.

The results indicated that average feed consumption,
final body weight and egg weight obtained during the final
2Q libitum feeding period (28 to 40 weeks of age) were not
significantly affected by previous feed restrictions per-
formed during either phases 1 or 2 or both. Thus, in the
long run, the net effect of restricted feeding followed
by ad libitum feeding may reside in the feed savings during
the feed restriction period without adversely affecting the
subsequent laying performance. Average egg weight, however,
increased more rapidly in groups which had previously re-
ceived a restricted amount of feed (treatments 3, 4 and
5) as compared with that of the control Ed libitum fed

group.

46

.moan mo mum

 

 

 

H
o.mm N.Nm m.om mmNH w.HOH m
H.mm v.mm m.Hh OONH m.mm v
H.>m m.Hm 0.0m ONNH m.v0H m
h.mm m.mm h.mh OONH H.v0H N
0.0m m.Nm o.vn mNNH m.NOH AHouucoov H
2:9 2:3 3; A53 33
ucoEummue
H.mx3 ovlvm H.mx3 emnmN H.mx3 ovumN H.mx3 ov H.mx3 ovaN
ucmHm3 mmo .m>¢ .coum mmo mediums .m>< .93 moon Hmchldm>¢ how\can\coom «mw¢

 

mqun
ocH>MH mo oUGMEHOMHmm map so mchoom EdanHH.mm he cmonHom mchoom wouoHuummu mo uommmo one .mH mHnme

. UOEHONMGMHU GuanHC

 

47

 

 

 

H
mv.mH on.vH om.mv m.ommov mN.mmH ov uouum
om.HH oo.vH om.mH H.mwvom vh.moH m m x 9
am .m 2. .s .3 .8 s . mmvm 3.05 m coﬂmoﬂmmm
on.HH Hm.o 00.50H m.ovth NN.wm v ucwEummue

em Hmuoe
.mxs ownvm .mxz vmnmN .mx3 oelmN .mxs oe .mg3 ovaN Eocooum GOHHmHum>
uanos mmm .coumlmmolwmplcmm w .u3 Noon Hmch mwc\©HHn\vmom mo momuoma mo oouaom

H
mumsom coo:

 

HHH ucoeHuwmxo cH mcHHn mcH>MH mo mocmauomuom on» you mocMHHm> mo mommHocﬂ .ON oHnoe

48

The results also indicated that birds fed Diet F
(treatment 2) were not significantly different from the
control as far as the average feed consumption, final body
weight and hen-day egg production were concerned.

The effect of two diets and restricted feeding followed
by ad libitum feeding on several liver characteristics
are shown in Tables 21 and 22.

The results indicated that previous feed restriction
during either the first (14 to 20 weeks of age) or the
second phase (20 to 28 weeks of age) caused significant
reductions in average liver wet weight, percent liver
fat, total liver fat, liver fat per kilogram of body
weight and incidence of liver hemorrhages at 40 weeks of
age as compared with corresponding values obtained in
the control ad libitum fed group.

When a group of birds which had previously received a
restricted amount of feed during phases 1 plus 2 (treat-
ment 5), those criteria mentioned above were not signifi-
cantly different from the control (treatments 5 vs. 1 in
phase 3). Furthermore, 16.7 percent of birds which had
received this treatment showed FLHS whereas none of the
FLHS occurred in the control or in other previously feed
restricted groups. Considering the fact that the average
percent liver fat of birds in treatment 5 was not very
high, the occurrence of FLHS might have happened by chance

alone.

‘49

.00.» ea: n.8uocnso 0» Dcdvuooud Houucoo loan .Ho.o v a. unduluudv aHucduHuHchwoo

OIIHCm “ ﬁg:

 

 

 

 

 

v
.vcilucouu £000 aHauH) one: no coaucoouomn
.uHunn a.) can
.uo>HH Gnu cH ace-cum noocnuuoao: mH can» auctH
..m 0.0Hn 5.0H o.mN b.0«m.n n.H«v.u H.o«n.nH o.n«n.nv m
m.n N.NHn o.o o.on cco.o«N.N acw.c«a.n coH.o«o.a och.H«v.mm v
N.m n.0Nn o.o H.o con.o«n.N c.u.c«H.v ccH.o«o.cH coo.H«N.om n
N.m v.an h.Hv o.cn coo.H«N.HH ccH.n«o.oH ecN.c«c.aN ocv.v«h.No N
H.n b.HNM , _ 0.0 o.cv o.o«o.c m.H«H.h H.o«H.mH vo.n«m.Nv .Houucou. H
..on. ..u.: .H-uu. . n... m... .au. .au. N... .90.
unusuqoue
nhnu h audH hlv\son Hw1on mnah GuanquIou .u) avon .ux\uoh you Hauoe can .u) out
am em-H noon \oxaueH auuoau nosma
aoHuuHuou

 

nuauonu uanH Houoaoo no ucH000u isanHH

mu ha ooroHHou ochoou couoHuuuou a can nUOHv or» no avenue one .HN oHnua

SC)

.mo.o v a a. acaumuacomm..
.mo.o v a u- ucaomuacumm.
. ngu Omaha cauoz

 

 

 

 

H

Nm.c no.mmmH Hn.NH oo.sn NH.oN NH.mm ov nouuu

ov.o oH.oamH wm.n mn.wH uH.mH Nw.cHH m m x a

mn.o Hn.mnom mH.cN no.oo «Nv.ma vv.oHN N coHueoHHmom

mw.H mv.Hnn ccca.moH acmm.vmv ccsm.osm ccom.mNnH v unusuooue

vm Hauoa
n>ev h uuoH cH umou anv\con\.u.z .Huox .93 Noon .ox\uuh uou Huuoe you p .93 um: socoouu coHuaHuu>
macaw. coo: Auo>quloumsmmnwmot «0 uoouuoo uo oousom

 

HHH unuBHuoaxo :« uoHuuHuouoaunso uo>HH Hnuoson u0u oocdHuo> uo nouaHoc< .NN oHaea

51

When birds were fed Diet F (treatment 2), average liver
wet weight, percent liver fat, total liver fat and liver
fat per kilogram of body weight were significantly higher
than corresponding values obtained from all other treat-
ments. The highest incidence of hemorrhages and fatty liver-
hemorrhagic syndrome type of hemorrhages (FLHS hemorrhages)
also occurred in birds that were fed Diet F.

In spite of the differences in liver characteristics,
energy intake and egg production measured during the last
seven days of the experiment were not significantly dif-
ferent between treatments (including Diet F). Thus, it
appeared that the level of energy intake did not seem to
be closely related to the production of high fat-liver or
FLHS under the conditions of this experiment.

Average final body weight or hen-day egg production
were not good indicators of FLHS of laying chickens, since
correlations between those criteria and liver characteris-
tics were very low (see Appendix Table 3).

High correlations between liver wet weight and the
percentage of liver fat (r = +0.85) and between the per-
centage of liver fat and total amount of liver fat (r =
+0.93) indicated that an increased liver wet weight was
partly due to an increase in the degree of fat deposition
which resulted in an increase in total fat in the whole
liver.

In this experiment the lowest total liver fat (TLF)

with FLHS hemorrhages was found to be 7.05 grams. Based

52

on the TLF of 7.05 grams, all birds used in this experiment
were divided into the following three categories in order
to compare the values of average feed consumption and liver
fat content in relation to presence or absence of FLHS
hemorrhages (Table 23).

Category A: Birds having more than 7.05 grams of TLF

with FLHS hemorrhages

Category B: Birds having more than 7.05 grams of TLF

without FLHS hemorrhages

Category C: Birds having equal to or less than 7.05

grams of TLF without FLHS hemorrhages

The data showed that out of 55 birds, 15 birds had
high liver fat values without having severe hemorrhages.
This indicated that although FLHS hemorrhages occurred in
birds with high liver fat values (TLF and percent liver
fat), a high level of fat in the liver was not always
related to the incidence of FLHS hemorrhages (A vs. B).

The only statistical difference in liver characteris-
tics measured between high fat-livers without having severe
hemorrhages and FLHS livers was liver wet weight. FLHS
livers were significantly heavier than high fat-livers
without having severe hemorrhages. This weight difference
could be due to the difference in water content, since
liver fat content between those two types of livers was not
significantly different.

During the course of this experiment, five birds died
of various diseases, however, no birds died of FLHS.

53

.umm» =u= m.ucmosum 0» maﬂmuooom Ho.o v m an unmoHMﬂcmam..
.ummu =u= m.ucmosum op mcwouooom mo.o v m um unmowMﬂcmﬂmg

.S.m.m H cam:

 

 

 

m
.Emsu ca
Aumm Hm>HH Hmuouv pow Em mo.h can» mmma mcﬂ>mn muw>HH cw H5000 no: cap mommnuuoﬁon mmqma
mm ma 5 mwuﬂn «0 “mnesz
.. Hm.oavs.bﬂ mm.o“-.mm oo.aﬂmm.am Acﬂmoumv any um>ﬂH w .m>a
.. . «.0“ m.em m.o“ H.mm v.HH «.5m isms .uz um: um>ﬂﬂ .m><
0.0“ o.ooa m.o“ o.hoa N¢.HH o.moa isms mmo\nuan\wmmm .m>a
mocmoﬂmacmﬂm so mo.h u may em mo.» A mas Ham mo.h A may
ucomnm ucmmnm ucwmmum
o .m> a m .m> a mu” m a

 

mumMQHHOEmn mmqm

 

mommnuuoamn mmqm mo mocmmnm Ho mocmmmnm
on coﬂumamu cw ucmusoo umm Hw>wa paw cowpmﬁdmaoo boom mmmum>m mo somwummﬁou .mm magma

D. Experiment IV

1. Procedure:

 

Experiment IV was conducted with 32-week old S.C.W.L.
pullets, which came from the same flock as the birds used
in experiment III.

Before starting this experiment all birds received the
same pullet developer diet and were also kept in the same
environmental temperature (15.90 C) from 14 to 22 weeks of
age as the birds utilized in experiment III. From 20 to
32 weeks of age, all birds were given a low-energy (Z-4)
laying ration. (Composition of Z-4 diet is listed in
Appendix Table 4.) Feed was given ad libitum from 14 to
32 weeks of age.

One of the two objectives of this experiment was to
test the effect of Diet F on FLHS. The other objective
was to study the effect of feeding regimes in combination
with different environmental temperatures on liver fat
accumulation and liver hemorrhage incidence.

During the experimental period of eight weeks, eight
birds in each of four treatments were held in individual
cages and were fed Diet F. The composition of Diet F may
be seen in Table 17.

In addition to pen 8-B at the Poultry Science Research

and Teaching Center, room S-A (air conditioned) in Anthony

54

55

Hall was utilized to take advantage of different environ-
mental temperatures. The temperature of room S-A had been
set to be 22.20 C, and electric lights in the room were
automatically turned on and off at 7:00 A.M. and 12:00
midnight, respectively.

During the four-week feed restriction period (32 to
36 weeks of age), birds in all treatments except treatment
1 (control, ad libitum) received a cyclic feeding treat-
ment (no feed every third day). From 36 to 40 weeks of
age, all birds were fed dd libitum.

Birds in treatment 1 received dd libitum feeding at
all times in 30.60 C (actual average temperature). Birds
in treatment 2 received a restricted feeding treatment in
30.60 C and were then fed ad libitum in 22.20 C (actual av-
erage temperature). Restricted feeding followed by 29
libitum feeding was performed in 30.60 C for treatment 3.
Birds in treatment 4 received a restricted feeding treat-
ment in 22.20 C and were then allowed to eat ad libitum
in 30.60 c.

Treatment description which includes environmental
temperatures during the feed restriction and the gd_;ibif
Egg feeding periods can be seen in Table 24.

Body weight was measured at the end of both the re-
stricted and the ad libitum feeding periods. Feed con-
sumption was measured every first and second day after

feed restriction (no feed) during the first four weeks,

56

Table 24. Treatment description, experiment IV

 

Age of birds
32-36 wks. 36-40 wks.
Treatment Feeding system 0C* Feeding system 0C*

 

1 (Control) §d_lib. 32.2 2§.ii§° 32.2
2 Restricted** 32.2 §d_lib. 22.2
3 Restricted** 32.2 E§.£i§' 32.2
4 Restricted** 22.2 3d lid. 32.2

 

*Environmental temperature; Actual average temperature
recorded for 22.20 C and 32.20 C
were 22.20 C and 30.60 C, re-
spectively.

**No feed every third day.

57

and it was measured every week during the ad libitum feeding
period of the second four weeks of experiment.

At the end of 40 weeks of age, birds in all treat-
ments were sacrificed for liver fat analysis.

Student's "t" test was used to compare FLHS livers
and high fat-livers without having severe hemorrhages, and

between FLHS livers and low-fat livers.

2. Results and Discussion:

 

The effects of feed restriction in different environ-
mental temperatures on average body weight and egg produc-
tion are shown in Tables 25 and 26.

Regardless of environmental temperatures employed,
birds in restricted feeding treatments were significantly
smaller in average body weights as compared with the control
dd libitum fed group, yet average hen-day egg production
was not affected by feed restriction.

When average body weights were compared within feed
restricted groups, birds kept in a lower environmental
temperature were heavier than birds kept in a higher
temperature. This was probably due to the increased feed
consumption.

The effect of restricted feeding followed by 29.l22£7
Egg feeding in different environmental temperatures on the
performance of laying birds can be seen in Tables 27 and

28.

58

.ummu

=u= m.uumccso o» mcﬂpuooom Honucoo Eoum Aao.o v my ucmummmwp >H¢GMUﬂMﬂcmwm««

.mnuﬂn mo «ma

 

 

m
.mmm mo mxom3 ozuuauuwaem
.Aoomm ocv coauowuumou comm mcw3oHHomH
o.Hm ««voma mmma ««¢.oma ««H.Mha v
m.>n «*mHmH mmma «am.hoa ««¢.hva m
H.vh ««wmma mmma ««m.moa «am.o¢H N
«.Hm mmba omma h.mm m.ooa AHouucoov H
Amy Asmv Aamv Aamv laws

usoﬁummue

m.mx3 mmlwm .mxs mm HMﬂuwcH amp cam Hmmp uma

 

.ooummmm Mmplcwz mm>¢

 

 

unmﬂm3 Noon .m>¢

puﬂn\pomm .m>¢

 

coapospoum mom can usmwm3 moon mmmuo>m no mmusu

nonmmEou Hmucoscouﬂ>cm ucmnmmm«p cw coauoﬁuummu comm mo muommmm one .mm manna

59

.Ho.o v a an uGMOAMAcmHm..
.mo.o v m um ucMUﬁMAcsﬂm.
.Aommm ocv coauowuummu comm mcHBOHHom

m
.UmEH0mmcmuu camond

 

 

H
mm.hm ~.naova m.ao~ o.mmv mm uouum
Ho.oq «m.omamm .«v.mama «.H.m~om m ucmEummua
aw Hmuoa

.mxz mmumm .mx3 mm sac can Name umH Eonmmnm aoﬂumﬂum>

 

 

coﬂuOSCOmemmm mmolcmm w unmwm3 moon Hmcﬂm pnﬁn\pmmm mo momumma mo oohnom
mumswm com:

H

 

>H ucmﬁwummxm cw coﬂumm cowuoﬂuum
no“ boom may mcwH5© woman mcﬂmwa mo mocmeuomumm on» How mocmwum> mo mmmaamcd .mm manna

60

.ummu =u= m.uumccsa op maﬂuuooom Houucoo scum Aao.o v as ucmumMMAu saucMUAMAcmﬂm«
.ummu =u. m.uumacso ou mcﬂouooom Houucoo scum Amo.o v as ucmummmeo mauamUAMﬁnmﬂm

a.

.1

 

 

m.mm mead .¢.mm a
H.~m .«momﬂ o.mm m
m.Ha Hana .«m.maa m
¢.mh Nana ~.vm Aaouuaoov a
2: A53 A53
.mxz ovuom .mxz ow .mx3 oqumm unusummua

 

 

 

.ooumvmmo mmolcma rm>¢ .u3 moon Hmcwm .m>¢ mmc\onﬂn\omom .m>¢

 

mpuﬂn
mcﬂamH mo mocmﬁu0mmWQ may no mousumummﬁou Hmucoﬁcouﬂ>cm unmummmwp

a“ mcﬂnmom asuﬁnﬂa am an cogoaﬂom ocﬂcmmm cmuouuummu mo powwow was .am manna

61

.Ho.o v m um UGMUHMﬂGOﬂmta
.mo.o v m an ucmowmacmﬂm.
. UmEHOM manna. ".1.an:

 

 

H

H
n¢.mm N.¢smma mo.bm mm nonum
mm.moa .w.aaa~m .«ma.omoa m ucmsummua

mm Hmuoe
.mxz oaumm .mxz ow .mxs ovumm aoommuu aoﬂumﬂum>

cowuosooum mom mooncom w .u3 moon Hmcﬂm wmp\ouﬂn\oomm mo mwmumma mo mousom
mnmowm cmmz

 

>H ucoawummxw ca oownmm mcﬂommm

Esuﬂnwa mm on» mcwuso woman mo mocmﬁuomumm map How mocmwum> mo mmmMHmcd .mm manna

62

At the same "high" environmental condition (30.60 C)
average final body weight of birds that received a restricted
feeding regime was significantly smaller than that of the
control dd libitum fed group. Average feed consumption
and hen-day egg production were not significantly dif-
ferent, however (treatments 3 vs. 1). These results along
with those obtained during the feed restriction period
indicate that body weight is more susceptible than the rate
of lay in response to reduction of feed intake.

When environmental temperature of the 3d libitum
feeding period was decreased, average feed consumption
increased significantly without affecting average final
body weight and hen-day egg production (treatments 2 vs.
1). The same phenomenon was observed during the dd_libdf
22m feeding period from the group which was previously
exposed to lower environmental temperature while feed re-
striction was performed (treatments 4 vs. 1). Therefore,
changing environmental temperature in either the feed re-
striction or the ad libitum periods may affect feed con-
sumption of birds during a later 29 libitum period.

The effect of two environmental temperatures and re-
stricted feeding followed by §d_libitum feeding on some
liver characteristics may be seen in Tables 29 and 30.

Average values for liver wet weight, percent liver
fat and total liver fat obtained from a group of birds

fed Z-4 diet were also included in Table 29 for reference.

6Z3

.u-o» .u. -.uuo::=a 0» unannouo- Houucou noun .na.o v a. acououudo aducauduucowm..
.uuou cu. u.uuoc:§o ou unavuoou- Hauucoo souu .mo.o v a. acouOuuuv auucau«u«co«mc
.!.u.m « coax.

.ucoauaouu non. c«nu«3 use: uo oo-ucoouom

 

 

.uauan an: no”
.uo>«~ on» :« use-cum oomuzquEon ma can» ouoza
.. .. o.o °.¢ -- o.« 5.. «.cn ..-N.
a.m .~.~on m.- m.~o H.H«a.a o.~«n.pa H.0«n.cn o.~ «a.mm .
m.m ....~ m.~H o.om o.~«..o w.~«u.n~ H.0«u..~ o.m «m.mm n
H.m ..v.mmn o.cm o.o°H ..H«n.a u.~«o.o~ ~.°«n.n~ n.. «o.ao N
«.m ..uo~ n.nn h.ou u.~«n.- o..«n.a~ m.°«a.o~ .n.°~«~.ho Aﬂouucooc H
..o:. ..u.x .H-ux. a... n... .26. .26. w... lea.
ucoEuooua
.a-o » an." unexcoa ouaauuoaog mags ooaauuoaoa .u: soon .oxxuam uau Hauoa you .u: an:

 

 

:« can” noun \oxuu=« suuonu H nosaa

 

unmanauouoouasu uo>ua
Hauo>oo so uc«000u Isuqnaa mm id optoenou UGHQIOu vouowuu-ou can nouauuuooaou Haucoscoua>cu 03» no uncuuo one .m~ canoe

 

64

.Ho.o v m an unmowmwcmwmgt

.wmauommcmuu Gauchd

 

H

mm.H mm.Nmm mo.HN mo.mm m®.om mm.amm 0N HOHHm
mm.o «amv.mmmb wh.mH ma.ov mn.®m om.~mm m unmﬁummha
mm Hmuoa

 

mama n ummH ca mmmm mmp\cm£\.m.z .HmoM .03 >©OQ .mx\umm pom Hmuoe pom w .93 #03 Eopomum coauMHHm>

H
mumswm cam: AH0>HHV mumsmm cum: mo mmmumoo «0 mousOm

 

>H quEwuomxm cw moﬂumauouowumno H0>HH Hmum>om mom wocmwuw> mo mmmhach .om manna

65

Although there were no significant differences in liver
characteristics between treatments, high fat-livers and some
of the FLHS livers were observed in all treatments.

A smaller total liver fat, and lesser incidence of
liver hemorrhages and FLHS, in most cases were noted
from groups of birds which had received restricted feeding
regimes in comparison to the control ad libitum fed group.

The only exception can be seen in treatment 2 where
birds received a restricted feeding treatment in 30.60 C
and then were allowed to eat ad libitum in 22.20 C. More
incidence of both liver hemorrhages and FLHS was observed
from the birds in treatment 2 than birds in the control.
This was probably due to a higher intake of energy of
birds in treatment 2 as compared with that of birds in the
control.

Higher incidence of liver hemorrhages was noted when
the environmental temperature during the feed restriction
and during the ad libitum period were different by 100 C
(treatments 2 and 4 vs. 3). Thus, switching environmental
temperature of the feed restriction and of the 3d libitum
period may affect incidence of liver hemorrhages.

As in the case of the previous experiment, FLHS hem-
orrhages occurred in livers with high fat but not all livers
with high fat contained FLHS hemorrhages (Table 31).

Average feed consumption, final body weight and hen-day

egg production between groups of birds having FLHS and

66

.ummp =u= m.ucmwsum 0» mcﬂuuooom Ho.o v m an unmoAMHcmﬁm..

.ummu =0: m.ucmosum ou mcﬂpuooom mo.o v m an unmowMﬂcmﬂma

.z.m.m H cow:

 

.Emnu ca Aumm Hw>wa Hmuouv umm Eb mH.mH can» mmma mcﬂ>mn mum>wa ca Haooo no: cap mommnuuoamn wank“
a ma m mcuﬂn mo umnauz

ms.oaam.vo on.oa~m.mo oo.o«m¢.mm Acumoumv .coum mom www-cos .m>«

ma amna ma sand v“ «aha isms .us soon Amado .m>¢

m.HH m.~0H m.o« m.mm m.HH m.koa isms amwxwuﬂnxwmmm .m>¢

. ¢>.o+me.m~ n¢.onm¢.¢m v5.0“mn.0m Anamoumc paw um>ﬂa a .m>«

.. . o.Hn m.m¢ m.on m.~m ~m.HH 0.05 lass .u3 um: um>ﬁa .m>¢

 

wocmoﬂmﬂcmﬂm as mH.~H.w mas so mH.~H A may em mH.~H A has

ucomnﬂ
U .m> m m .m> < 0

H
Hammad ucmmoum

m d

 

mommnunoaon mmqm

 

mommauuoamn mmqm mo oncomnm

uo mocmmmum ou coﬂumHou ca mpnﬂn mo mocmauomuom 0:» van ucmucoo pom Hm>wa mo cemﬁummEou

.Hm mHQMB

67

high liver fat content without massive hemorrhages, and
between birds having FLHS and low liver fat content were
not significantly different (Table 31).

Although the highest incidence of hemorrhages and
FLHS occurred in the particular group of birds which con-
sumed the highest energy (treatment 2), energy intake in
general did not appear to be significantly related to liver
characteristics as indicated by low correlations between
these variables (Appendix Table 5) and by feed consumption
comparison data of Table 31.

In this experiment, no mortality due to FLHS was re-
corded (2 birds died of other diseases).

Considering the fact that high fat-liver and FLHS
liver occurred predominantly in the group of birds fed
Diet F in experiment III, and that high fat-liver and some
incidence of FLHS occurred in all treatments of experi-
ment IV in which only Diet F was used, a possibility ex-
ists that some factor(s) in Diet F is/are involved in pro-

ducing high fat-liver, if not FLHS.

V. GENERAL DISCUSSION

A review of the literature indicated that there are
different opinions concerning the relationship between
dietary energy level and occurrence of fatty liver in
chickens. Several investigators have reported that an in-
creased dietary energy level caused an increase in liver
fat, while others did not observe this relationship. At
any rate, it may not be reasonable to try to relate a low
energy diet and high incidence of fatty liver problem un-
less some chemical substances which are toxic to the liver
cell are present in the diet.

Little information was available on the performance
of chickens and liver fat content as influenced by dietary
energy sources. Thus, it seemed desirable to study the
effect of high energy diets of different energy sources
on the performance and liver fat accumulation of chickens
for the first experiment.

The results of experiment I showed that when birds
were fed the diet that contained herring oil as a primary
energy source, those birds consumed the least amount of
feed and produced fewer eggs than did the birds fed all
other isocaloric-isonitrogenous diets. This phenomenon

was also observed in experiment II, where environmental

68

69

temperature (27.80 C) was higher than that of experiment

I (12.20 C). Thus, it appeared that although birds are
known to have few taste buds, apparently the taste of
herring oil diet (or some other factor(s) in the feed)

was not desirable for them and as a consequence a reduc-
tion of feed consumption resulted. Decreased egg produc-
tion appeared to be due to a marked decrease in feed intake
unless some other factor in the herring oil diet influenced
egg production.

The results also indicated that in either environmental
temperature employed energy from carbohydrate sources was
not statistically different from that of fat sources on
their respective influence on liver fat content. Slightly
higher percent liver fat, however, was obtained from birds
fed a high carbohydrate diet in a "high" environmental
temperature (27.80 C). In general, livers from laying
birds fed the same diets but kept in a "high" environ-
mental temperature had a higher percentage of liver fat
and a higher incidence of liver hemorrhages than laying
birds kept in a "moderate" (12.20 C) environmental tem-
perature. Similar phenomenon has been previously ob-
served by Nesheim 2E.2l' (1969). Direct comparison between
the two experiments in these trials, however, cannot be
made due to different experimental conditions.

The definition of FLHS liver is rather subjective.

If FLHS livers were defined as those livers which were

70

"fatty" (15 percent or more fat on wet basis) and contained
more than 15 hemorrhages and/or hematomas, none of the
diets used in experiments I and II caused FLHS.

According to Masoro (1968) the amount of fat in the
liver is influenced by the balance of the input processes
of triglyceride precursors (from dietary carbohydrate and
fat) into the liver for formation of triglyceride and the
processes of removing triglyceride from the liver (output
processes). There are two known output pathways. One is
that the triglyceride is incorporated by the endoplasmic
reticulum into the lipoproteins and secreted in this form
into the plasma. The other pathway is that triglyceride
undergoes lipolysis to free fatty acids and glycerol, which
can be oxidized in the mitochondria.

In experiment III, when birds were fed a restricted
amount of feed, average body weight was significantly
smaller than the control dd libitum fed group. Average
egg production, however, was not different from the
control group. If one hypothesizes that the rate of hepatic
lipid oxidation is constant and lipid going out into the
yolk is much greater than that going out for the formation
of body fat, one may expect to observe a smaller percentage
of liver fat as well as a smaller liver wet weight of birds
in a restricted feeding group.

When birds were allowed to eat ad libitum after termi-

nation of either the first (14 to 20 weeks of age) or the

71

second phase (20 to 28 weeks of age) of restricted feeding,
their average liver wet weights and the percentages of liver
fat were significantly smaller than those from birds that

had no previous experience of receiving a restricted feeding
regime. However, average feed consumption, final body weight
and percent hen-day egg production during the final ad
libitum period (28 to 40 weeks of age) were not significantly
different from the birds that never had experienced re-
stricted feeding. If feed intake for those previously

feed restricted groups had been greater than the control
group the liver fat content would be similar to that of the
control, but since feed intake was not different from the
control (along with body weight and egg production), liver
fat content probably did not increase much after the time
period when feed restriction was performed. Thus, percent
liver fat remained at a low level.

While comparing a low (2359 Kcal. M.E./kg.) and a high
energy diet (same ration as Diet F, 3004 Kcal. M.E./kg.),
Barton (1967) observed fatty liver and liver hemorrhages
(13 percent) only in the group of birds that were fed the
high energy diet. Thus, he concluded that fatty liver
incidence was higher among hens fed a diet containing
high level of energy.

In experiment III, however, when a group of birds
was given Diet F 2Q libitum, which was 130 kilocalories

(M.E./kg.) lower than the control diet, a marked increase

72

in the average liver wet weight, liver fat (both the per-
centage and total), and FLHS resulted. This may indicate
that a high level of dietary energy pg£_§g is not the major
factor in producing high-fat liver or FLHS.

Wolford and Polin (1972a) reported that when hens were
force-fed, the degree of liver fat accumulation and inci-
dence of FLHS were linear to the amount of feed received.
Their result was probably due to increased input, which
led to increased formation of triglyceride in the liver,
and little change in output of lipid in the form of egg
production. However, it appears that an increase in
feed (energy) intake is not the only way to produce FLHS
because in experiment III, birds fed Diet F did not consume
more calories than those in the control or in other treat-
ments, yet the majority of birds having high fat-liver
and FLHS were observed in this (Diet F) treatment group.

The relationship between high fat-liver and FLHS was
also studied in experiments III and IV. The results
showed that FLHS hemorrhages occurred in livers with high
fat (high fat-liver), yet not all high fat-livers contained
FLHS hemorrhages. This indicated that the cause of high
fat-liver and FLHS may not necessarily be the same. A
similar observation was made by Wolford and Polin (1972b).

In experiment IV, the highest incidence of FLHS hemor-
rhages resulted in birds that first received a restricted
feeding regime in a "high" environmental temperature

(30.60 C) and then received an dd libitum feeding in a

73

reduced temperature (22.20 C). Thus, changing the environ-
mental temperature coupled with a restricted-dd libitum
feeding regime may affect the incidence of liver hemor-
rhages. However, further studies are needed to elucidate
the exact cause(s) of liver hemorrhages.

Couch (1956) and Reedy (1968) characterized fatty liver
syndrome as increased body weight and decreased egg pro-
duction. In these experiments, however, birds having
FLHS were neither heavier nor poorer egg producers than
the control group. Ringer and Sheppard (1963) also re-
ported that egg production rate was not affected by the
fatty liver condition. Therefore, body weight and egg pro-
duction may not be used as satisfactory indications of

FLHS in laying birds.

VI . SUMMARY

A series of 11 diets were used in the first two ex-
periments to study whether various sources of energy ex-
hibited different effects on liver fat accumulation.

In isocaloric-isonitrogenous diets, different sources
of carbohydrate and fat as primary energy supplements
were not different in exerting their effect on liver fat
accumulation in the "moderate" (12.20 C) environmental
temperature. When the environmental temperature was "high"
(27.80 C), more livers appeared to be fatty, and the in-
cidence of liver hemorrhages was greater than those ob-
tained in the "moderate" environmental temperature.

Average feed consumption of birds fed diets that were
calculated to be isocaloric and isonitrogenous but formu—
lated from different dietary ingredients was not the same.
This implies that there are some factor(s) in the feed in-
gredients other than dietary energy which may influence
feed consumption.

In one experiment, birds were given a restricted amount
of feed for a certain period of time. Percentage of liver
fat as well as total amount of liver fat of birds that re-
ceived the restricted feeding regime was lower than the

control ES libitum fed group.

74

75

When Diet F, a commercial laying formula, was fed
ad libitum, liver wet weight, percent liver fat and total
liver fat were markedly increased and some incidence of
fatty liver-hemorrhagic syndrome was observed. However,
average feed consumption, final body weight and percent
hen-day egg production were not significantly different
between treatments.

In another experiment feed restriction and 3d libitum
feeding were practiced consecutively under two different
environmental temperatures to study their effect on liver
fat accumulation and liver hemorrhaging. Once again fatty
liver and some incidence of FLHS were observed among all
treatments in which Diet F was used.

The highest amount of total liver fat was observed in
the control group which received dd libitum feeding in a
"high" (30.60 C) environmental temperature.

Total liver fat as well as the incidence of both the
liver hemorrhages and FLHS were the lowest among birds
that received a restricted feeding regime in the "high"
environmental temperature. The highest incidence of
liver hemorrhages and FLHS occurred among birds that first
received a restricted feeding regime in 30.60 C and then
were allowed to eat feed dd libitum in 22.20 C.

This indicated that feeding regime along with environ-
mental temperature might affect liver hemorrhages.

FLHS hemorrhages occurred in livers with high fat

values but not all livers with high fat exhibited FLHS

76

hemorrhages. FLHS livers were significantly heavier than
livers that did not have severe hemorrhages.

Average feed consumption, final body weight and hen-
day egg production between groups of birds having FLHS

and low liver fat were not significantly different.

LITERATURE CITED

VII . LITERATURE CITED

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Best, C. H., J. M. Hershey and M. E. Huntsman, 1932a. The
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Best, C. H. and M. E. Huntsman, 1932b. The effects of
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Bull, H. S., 1968. Fatty liver syndrome in laying hens.
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Chalvardjian, A., 1966. Mode of action of choline I.
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Chalvardjian, A., 1969. Mode of action of choline II.
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Couch, J. R., 1956. Fatty livers in laying hens. A con-
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Couch, J. R., 1964. The fatty liver syndrome in laying
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Deacon, L. E., 1968. The fatty liver syndrome--History
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Donovan, G. A. and S. L. Balloun, 1955. Effect of dietary
protein and energy on chick liver fat accumulation.
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Dudley, W. A., L. J. Machlin and G. J. Marco, 1961. The
effect of dietary vanadium and cholesterol on the
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Duke, M. J., R. K. Ringer and J. H. Wolford, 1968. Failure
of plasma protein level to indicate developing fatty
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Du Vigneaud, V., J. P. Chandler, M. Cohn and G. B. Brown,
1940. The transfer of the methyl group from methionine
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Fisher, H. and H. S. Weiss, 1956. Observation on the fatty
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Griffith, M. and R. Schexnailder, 1972. Evaluation of cho-
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Hamilton, P. B. and J. D. Garlich, 1970. Aflatoxin as a
cause of fatty liver syndrome in laying hens. Poultry
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Hamilton, P. B., H. T. Tung, J. R. Harris, J. H. Gainer
and W. E. Donalelson, 1972. The effect of dietary fat
on aflatoxicosis in turkeys. Poultry Sci. 51: 165-170.

Harper, A. E., W. J. Monson, D. A. Benton, M. E. Winje and
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Hegsted, D. M., R. C. Mills, C. A. Elvehjem and E. B. Hart,
1941. Choline in the nutrition of chicks. J. Biol.
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79

Howes, J. R. and T. C. Fitzgerald, 1966. Fatty livers in
coturnix quail fed high dietary levels of fat. Poultry
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Jensen, L. S., G. W. Schumaier, A. D. Funk and T. C. Smith,
1970. A new lipotropic agent for the laying hen.
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Kirk, R. E., 1968. Experimental design: Procedures for
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Leach, R. M., Jr., E. Ceballos and M. C. Nesheim, 1966.
Recent studies on choline requirements of chicks and
hens. Proc. Cornell Nutr. Conf. pp. 69-72.

Leveille, G. A., 1967. In vivo fatty acid synthesis in
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Leveille, G. A., 1969. In vitro hepatic lipogenesis in the
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Lorenz, F. W., C. Entenman and I. L. Chaikoff, 1937-1938.
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Lorenz, F. W., I. L. Chaikoff and C. Entenman, 1938. Liver
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80

Nelson, F. E., 1968. Effect of inositol on liver fat of
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81

Reed, J. R., L. E. Deacon, F. Farr and J. R. Couch, 1968.
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39: 1550-1555.

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Weiss, H. S. and H. Fisher, 1957. Plasma lipid and organ
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Wolford, J. H. and D. Polin, 1972a. Laboratory duplication
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APPENDIX

82

Composition of vitamin-trace mineral
premixture for laying ration (Diets 1
to 11)

Appendix Table l.

 

Micronutrients Per kilogram of diet

 

Vitamin A, I.U. 10208.00
Vitamin D3, I.C.U. 3190.00
Riboflavin, mg. 8.93
Pantothenic acid, mg. 15.31
Niacin, mg. 31.90
Choline chloride, mg. 497.64
Folic acid, mg. 1.28
Vitamin B12, ug. 15.31
Vitamin E, I.U. 6.38
Menadione sodium bisulfite, mg. 1.91
Manganese, mg. 71.98
Iodine, mg. 1.12
Copper, mg. 4.54
Cobalt, ug. 252.00
Zinc, mg. 56.00
Iron, mg. 28.10
Magnesium, mg. 580.00

 

83

Appendix Table 2. Composition of pullet developer diet

 

 

 

Ingredient Percent

Ground yellow corn 50.00
Soybean, 49% protein 3.13
Ground oats 25.07
Wheat standard, middlings 10.00
Alfalfa meal, 17% protein 3.00
Fishmeal, 49% protein 2.00
Meat & bone meal, 50% protein 3.50
Dicalcium phosphate 1.00
Ground limestone 1.50
Salt, iodized 0.25
Manganese sulfate 0.01
Vit.-min. premix* 0.50
Methionine 0.04

Total 100.00

Calculated analysis
Kcal. M.E./kg. diet 2809.3

Crude protein, % 14.0

 

*Supplied the following per kg. of diet: Vitamin A, 6600
I.U.; Vitamin D , 2200 I.C.U.; Riboflavin, 4.08 mg;
Pantothenic aci , 6.60 mg; Niacin, 19.80 mg; Choline
chloride, 440 mg; Vitamin B12, 9.92 ug; Menadione sodium
bisulfite, 1.68 mg; Vitamin E, 1.68 I.U.; Manganese,

64.0 mg; Iodine, 1.00 mg; Copper, 4.0 mg; Cobalt, 249.6
pg; Zinc, 50.0 mg; Iron, 25.0 mg.

84

Appendix Table 3. Correlation coefficients of the variables
obtained during the final ad libitum feeding
period (28-40 weeks of ageT_of experiment
III

 

Variables Liver wet wt. % Liver fat Total liver fat

 

Feed consumption 0.165 0.109 0.092
Energy intake 0.099 0.045 0.025
Final body wt. 0.182 0.158 0.095

Hen-day egg prod. 0.130 0.113 0.124

 

85

Appendix Table 4. Composition of Z-4 laying formula

 

 

 

Ingredient Percent
Ground yellow corn 34.5
Ground oats 20.0
Wheat bran 15.0
Flour middlings 10.0
Alfalfa meal, 17% protein 3.0
Meat scraps 3.0
Dried skim milk 2.0
White fish meal, 60% protein 2.5
Soybean meal, 44% protein 2.5
Ground oyster shell flour 5.0
Steamed bone meal 1.5
Salt, iodized 0.6
Cod liver oil (400 vitamin D, 2000 vitamin A) 0.4
Total 100.0
Calculated analysis
Kcal. M.E./kg. diet 2359
Crude protein, % 14.8

 

86

Appendix Table 5. Correlation coefficients of the variables
obtained during the dd libitum feeding
period (36-40 weeks of age) of experi-

 

 

ment IV
Variables Liver wet wt. % Liver fat Total liver fat
Feed consumption 0.172 -0.211 -0.082
Energy intake 0.115 -0.180 -0.078
Final body wt. 0.305 0.100 0.263

Hen-day egg prod. -0.104 0.110 0.073