J . Sci. Food Agric. 1981,32, 1047-1056

Identification of the Component Responsible for the Effects of Added Dietary Copper Sulphate in the Female Domestic Fowl

Norman Jackson and Mary H. Stevenson

Agricultural arid Food Chemistry Research Division, Department of Agriculture for Northern Ireland and the Queen's University of Belfast, Newforge Lane, Belfast BT9 5PX

(Manitscript received 25 November. 1980)

Sulphate, as sulphuric acid, was fed to Shaver 288 and Warren Studler SSL laying hens for 280 days at levels equivalent to 0.2, 0.4, 0.6, 0.8 and 1.0 g sulphate kg-l diet. The results obtained were compared with those found using similar diets to which the sulphate was added as copper sulphate pentahydrate (CuSO4. ~ H z O ) . Addition of HzS04 had no effect on food intake, food conversion efficiency, body-weight or egg production. The addition of CuSO4 caused adverse effects on food intake, egg production and body-weight. The HzS04 had no effect on liver, kidney, oviduct, ovary and gizzard weight per unit body-weight. CuSO4 decreased ovary and oviduct weights and increased gizzard weight. HzS04 had no significant effect on liver copper (Cu) concentrations but CuSO4 increased liver Cu. The highest level of CuSO4 addition depressed liver lipid concentration in the Shaver 288 birds. Both the HzS04 and the CuSO4 affected the levels of individual fatty acids.

1. Introduction

The growth promoting effects of copper compounds in the pig are considered to be due to the presence of the Cu rather than the anion.', Copper compounds other than the sulphate have also shown positive growth responses in chicks 3-G and turkey po~l t s .7-~

Gordon and Sizer'O suggested that, in the chick, it was the sulphate ion rather than the Cu ion which was the growth promoting moiety of copper sulphate (CuS04.5HzO) but this view was not supported by Coates and Harrison.11

Evidence suggests that sulphate can exert a sparing effect on sulphur-containing amino acids in poultry diets.l2-l8

Effects on poultry carcass quality as a result of giving Cu-supplemented diets are limited. No significant changes in the fatty acid composition of the subcutaneous fat of chicks have been rep0rted.1~2 Changes in the fatty acid composition of liver fat and the fat adhering to the gizzard of laying hens given dietary Cu compounds have been observed but they could not be specifically associated with the additives.21

Effects of CuSO4 and cupric oxide (CuO) on the female domestic fowl have been studied in this lab0ratory.~1-2~ Many of the detrimental effects previously attributed to CuSO4 have been found to be the result of appetite depression.26 The present experiment was designed to attempt to separate the effects of Cu and of sulphate.

2. Experimental

The experiment commenced on 15 May 1979 and continued for ten 28-day periods until 18 February 1980. Two hundred and sixty four 18-week-old laying hens previously vaccinated against Marek's disease, infectious bronchitis and epidemic tremors and comprising 132 Shaver 288 and 132 Warren Studler SSL birds were used. The battery house was unheated, the minimum recorded winter

0022-5142/81/1100-1046 $02.00 0 1981 Society of Chemical Industry 1047

70

1048 N. Jackson and M. H. Stevenson

temperature being 4°C and the maximum summer temperature 25°C. The lighting regimen was 15 h light and 9 h darkness. At 27 weeks of age, the hens were randomly allocated to one of 12 treatment groups giving 11 hens of each breed per treatment. The diets, fed ad /ib., were the control dietz2 and this diet with 0.2, 0.4, 0.6, 0.8 or 1.0 g sulphate kg-1 added either as sulphuric acid (HzS04) (in water, 1.25 litres 100 kg-1) (treatments 2-6) or as finely ground CuSOa.5HzO2z (treatments 7-11). Water without additional HzS04 was added to all the other diets. The control diet contained (kg-1): 866.7 g dry matter, 158.9 g crude protein (CP) (N x 6.25), 10 mg Cu, 128 mg Zn, 296 mg Fe, 31.9 g Ca, 5.2 g P and had a calculated apparent metabolisable energy (AME) content of 11.4 MJ kg-l. The levels of sulphate added as CuSO4 were equivalent to 150, 300,450, 600 and 750 mg Cu kg-1 diet. A further treatment was included (treatment 12) in which 1.5 g m-niethionine kg-l was added to the control diet.

Eggs were recorded daily and weighed twice weekly. Hens were weighed initially and at the end of each 28 day period and mean body-weight obtained from these observations. Food intake was measured for each 28 day period. After 40 weeks, four birds of each breed per treatment were randomly selected and killed by decapitation. Blood serum Cu was determined.Z7 Liver, kidneys, oviduct, ovary and gizzard were removed and weighed. The inorganic sulphate content of the diets was measured.Zs Other chemical and statistical analyses were carried out as described previously by Jackson and Stevenson.21

3. Results

Mortality was low in this experiment (2.3 %) and the deaths were equally distributed between birds consuming the CuSO4- and the HzS04- supplemented diets. Of the dead birds 86% were Shaver 288.

Mean food intake, egg number, mean ‘period’ mean and total egg weight and food conversion efficiency of the hens are given in Table 1. The addition of the methionine had no significant effect on any of these parameters. Overall food intake, egg number, mean and total egg weight and food conversion efficiency were all significantly affected (P < 0.001) by the additive. Food intake showed a quadratic relationship (Ik0.05) with level of additive while egg number, mean and total egg weight, and food conversion showed a negative linear response with level of additive (all P<O.OOl). The interaction of form of additive x level of additive with food intake was significant (P<O.001), its effect being mainly concentrated in the significantly differing linear responses (P< 0.001) within the two forms of sulphate. Differing quadratic responses were shown by egg number (P < 0.01), total egg weight and food conversion (both P<0.05) within the forms of sulphate. It can be seen (Table 1) that the differing responses within the forms of sulphate arise through no significant effect between levels of HzS04 compared with a significant decrease in the variables at higher levels of CuSO4.

Table 2 shows initial and final body-weights, mean body-weight, daily crude protein and meta- bolisable energy intakes. Breed had a significant effect on initial body-weight, the mean initial body-weight for the Shaver 288 and Warren Studler birds being 1.52 and 1.86 kg, respectively. The form of additive significantly affected final and mean body-weight (both P<O.OOl), both of which showed a negative linear response (P< 0.001) to level of additive and differing linear responses within the forms of additive (P<O.Ol and < 0.001, respectively).

The fresh weights of liver, oviduct, ovary and gizzard (expressed as kg-l body-weight) are given in Table 3. Overall, when liver weight was expressed as g kg-1 body-weight, both form and level of additive produced no significant effect although there was a negative linear relationship ( P < 0.05) with level of additive. Oviduct weight per unit body-weight was affected by form of additive (P< 0.01) and the overall effect of level of additive was not significant, although there was a negative linear response to level of additive (P<O.Ol) . Ovary weight per unit body-weight was reduced by form (P< 0.01) and level of additive (P<O.O5), the response to the latter being linear ( P < 0.01). Gizzard weight kg-1 body-weight showed a linear increase with level of additive (P< 0.01) but differing linear responses within the two forms of additive ( P < 0.001). All these significant effects on tissue weights are due to the marked effects produced by the added CuSO4. There was an absence of res- ponse to the dietary HzS04. Kidney and thyroid gland weights kg-1 body-weight were unaffected

Tab

le 1

. M

ean

daily

foo

d in

take

, egg

num

ber,

‘pe

riod

’ mea

n an

d to

tal e

gg w

eigh

t and

food

con

vers

ion

of l

ayin

g he

ns g

iven

HzS

O4-

, CuS

Oa-

and

met

hion

ine-

supp

lem

ente

d di

ets

for 4

0 w

eeks

Bre

ed

and

addi

tive

0

HzS

04

33.1

7 cu

so4

Sh

aver

War

ren

Has

04

34.3

5 St

udle

r C

uSO

a

HzS

04

239

cuso

4

Shav

er

War

ren

HzS

Os

224

Stud

ler

CuS

O4

HzS

04

61.7

cu

so4

Sh

aver

War

ren

HzS

Os

6,.8

St

udle

r C

uSO

a

H2S

04

14.7

1 Sh

aver

cu

so4

War

ren

HzS

04

13,7

5 St

udle

r C

uSO

a

HzS

04

0.44

6 Sh

aver

cu

soa

War

ren

His

04

o.

399

Stud

ler

CuS

O4

Leve

l of

addi

tive=

(g

so4

kg-l

)

0.2

0.4

0

.6

0.8

___

1 .o

Met

hion

ine

(g k

g-9

1.5

Stat

istic

al si

gnifi

canc

e of

eff

ect

Form

of

Leve

l of

Form

of

addi

tive

x R

espo

nse

Bre

ed

addi

tive

addi

tive

leve

l of

addi

tive

Foo

d in

take

(kg

) 33

.09

33.4

3 33

.66

33.8

2 34

.30

34.2

0 27

.11

23.2

2 35

.22

35.5

6 35

.60

35.5

2 34

.85

34.0

0 31

.35

23.8

8

Egg

num

ber

23 1

212

23 1

24

4 23

0 24

2 16

3 11

4 22

2 22

5 20

5 21

5 24

0 23

0 21

0 16

7

Mea

n ‘p

erio

d’ m

ean

egg

wei

ght (

g)

62.3

62

.8

61.3

59

.8

60.2

59

.2

58.4

58

.2

62.9

60

.5

62.4

61

.0

60.7

60

.4

58.8

59

.6

Tota

l egg

wei

ght (

kg)

14.3

3 13

.28

14.1

8 14

.51

13.8

1 14

.33

9.51

6.

78

13.8

3 13

.51

12.6

7 12

.97

14.5

0 13

.74

12.2

4 9.

96

Foo

d eo

nver

sion

(kg

egg

s kg-

’ fo

od)

0.43

2 0.

398

0.42

1 0.

431

0.40

1 0.

421

0.34

5 0.

285

0.39

3 0.

382

0.35

5 0.

366

0.41

8 0.

407

0.39

0 0.

421

33.0

5 19

.83

35.2

3 21

.03

217 88

24

0 10

9 61.3

56

.3

60.2

56

.5

13.2

8 4.

96

14.3

7 6.

20

0.40

1 0.

247

0.40

9 0.

292

32.6

2

34.5

4

209

24 I 61 .O

61.5

12.8

4

14.7

7

0.38

6

0.42

9

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

***

***

***

*** *

S.e.

m. 0

.986

* **

* **

* **

* N

S S.

e.m

. 10

.9

***

NS

* ***

NS

S.e.

m.

1.25

***

***

***

NS

NS

S.e.

m. 0

.696

***

***

***

NS

NS

S.

e.m

. 0.0

217

***

***

NS ***

*** **

NS

NS

NS ***

+** * ***

*** *

a T

he le

vels

of

adde

d C

u co

rres

pond

ing

to t

he le

vels

of

adde

d su

lpha

te a

re 0

, 15

0, 3

00,4

50,

600,

and

750

mg

Cu

kg-1

.

* Per

iod

mea

n eg

g w

eigh

t is:

tota

l wei

ght

of e

ggs/

peri

od

num

ber

of e

ggs/

peri

od .

NS

=N

ot s

igni

fica

nt; *

P<O

.O5,

** P

<O.O

l,-*

**

P<O

.OO

I.

Val

ues

are

the

mea

n of

11

obse

rvat

ions

.

Tabl

e 2.

Mea

n in

itial

and

fin

al b

ody-

wei

ghts

, m

ean

body

-wei

ght,

daily

cru

de p

rote

in a

nd a

ppar

ent

met

abol

isab

le e

nerg

y in

take

of

layi

ng h

ens

give

n H

zSO

d-,

CuS

O4-

and

m

ethi

onin

e-su

pple

men

ted

diet

s fo

r 40

wee

ks

Leve

l of

addi

tives

St

atis

tical

sig

nific

ance

of

effe

ct

Bre

ed

(g so

4 kg

-l)

Met

hion

ine

and

(g k

g-9

Form

of

Leve

l of

Form

of

addi

tive

x ad

ditiv

e 0

0.2

0.4

0

.6

0.8

1 .o

1.5

R

espo

nse

Bre

ed

addi

tive

addi

tive

leve

l of

addi

tive

HzS

O4

cuso

4

Shav

er

War

ren

HzS

04

Stud

ler

CuS

O4

HzS

O4

Shav

er

CuS

O4

War

ren

HzS

Oa

S tud

ler

CuS

O 4

War

ren

HzS

04

Stud

ler

CuS

O4

Shav

er

HzS

04

cUso

4 W

arre

n H

zS04

St

udle

r C

uSO

4

HzS

04

cuso

4 Sh

aver

War

ren

HzS

04

Stud

ler

CuS

Oa

1.50

1.84

1.76

2. I6

1.65

2.02

18.8

19.5

1.35

I .40

Initi

al b

ody-

wei

ght

(kg)

1.

51

1.52

1.

52

1.51

1.

51

1.53

1.

52

1.53

1.

53

1.52

1.

88

1.83

1.

87

1.86

1.

84

1.87

1.

83

1.87

1.

88

1.86

Fina

l bod

y-w

eigh

t (kg

) 1.

70

1.72

1.

76

1.68

1.

65

1.73

1.

59

1.62

1.

54

1.50

2.

15

2.09

2.

19

2.05

2.

14

2.16

2.

16

1.91

1.

86

1.78

Mea

n bo

dy-w

eigh

t (kg

) 1.

62

1.63

1.

69

1.60

1.

61

I .66

1.

59

1.55

1.

46

1.44

2.

06

1.99

2.

06

1.98

2.

04

2.03

2.

02

1.87

1.

75

1.65

Dai

ly c

rude

pro

tein

inta

ke (

g)

18.8

19

.0

19.1

19

.2

18.8

19

.5

19.4

15

.4

13.2

11

.3

20.0

20

.2

20.2

20

.2

20.0

19

.8

19.3

17

.8

13.6

11

.9

Dai

ly a

ppar

ent

met

abol

isab

le e

nerg

y in

take

(M

J)

1.35

1.

36

1.37

1.

38

1.34

1.

40

I .39

1.

10

0.95

0.

81

1.43

1.

45

1.45

1.

45

1.43

1.

42

1.38

1.

28

0.97

0.

86

1.53

I .87

1.71

2.10

1.63

2.00

18.5

19.6

1.33

1.41

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

***

NS

NS

N

S

NS

S.e.

m. 0

.050

***

***

**

***

NS

S.e.

m. 0

.071

***

***

***

***

NS

S.

e.m

. 0.

057

***

***

***

*** *

S.e.

m. 0

.560

***

***

***

*** *

S.e.

m. 0

.040

NS

NS

N

S * **

NS **

***

NS ***

***

NS

**a

***

NS

a T

he le

vels

of

adde

d C

u co

rres

pond

ing

to th

e le

vels

of

adde

d su

lpha

te a

re 0

, 15

0, 3

00, 4

50,

600

and

750

mg

Cu

kg-1

. N

S=

Not

sig

nifi

cant

; * P<0

.05,

** P

<O

.OI,

*** P

<O.O

OI.

V

alue

s ar

e th

e m

ean

of 1

1 ob

serv

atio

ns.

Tab

le 3

. M

ean

fres

h w

eigh

ts (

g kg

-l bo

dy-w

eigh

t) of

live

r, ov

iduc

t, ov

ary

and

gizz

ard

of la

ying

hen

s gi

ven

HzS

O4-

, CuS

O4-

and

met

hion

ine-

supp

lem

ente

d di

ets

for

40 w

eeks

Leve

l of

addi

tive"

St

atis

tical

sig

nific

ance

of

effe

ct

Bre

ed

(g so

4 kg

-l)

Met

hion

ine

and

(g k

g-9

Form

of

Leve

l of

Form

of

addi

tive x

ad

ditiv

e 0

0.2

0.4

0.6

0.8

1 .o

I .5

Res

pons

e B

reed

ad

ditiv

e ad

ditiv

e le

vel o

f ad

ditiv

e

Shav

er

War

ren

Stud

ler

Shav

er

War

ren

Stud

ler

Shav

er

War

ren

Stud

ler

Shav

er

War

ren

Stud

ler

Live

r (g

kg-

1 bo

dy-w

eigh

t)

23.4

20

.8

20.4

20

.3

17'0

21

.9

20.1

19

.9

20.5

17.5

20

.3

11.5

18

.9

16.8

17

.0

18.0

:::: O

vidu

ct (

g kg

-1 b

ody-

wei

ght)

40

.8

33.6

35

.8

35.2

36

'1

24.4

37

.4

26.3

23

.4

29.1

28

.0

25.1

28

.0

27'9

29

.5

26.2

27

.1

12.2

Ovn

ry (g

kg-

l bo

dy-w

eigh

t)

28.3

19

.6

23.4

28

.1

24.6

20

.1

24.2

16

. I

16.0

23

.3

19.6

24

.1

23.7

22

.9

20.7

19

.6

8.8

16.4

Giz

zard

(g

kg-'

body

-wei

ght)

15

.7

14.3

16

. I

14.9

13

'3

13.5

14

.1

13.1

16

.8

12.3

11

.6

12.5

11

.2

12.2

10

.2

11.1

12

.3

15.2

16.9

19

.0

15.4

17.7

17

.3

17.6

27'9

28

.6

15.6

28

.5

20.0

30

.2

22.4

16

.1

12.2

22.2

17

.1

12.0

14.2

15

.6

16.1

10

.9

15.4

10

.3

Ove

rall

NS

NS

Lin

ear

Qua

drat

ic

Ove

rall

* **

L

inea

r Q

uadr

atic

Ove

rall

14s

**

Lin

ear

Qua

drat

ic

NS

***

Ove

rall

Lin

ear

Qua

drat

ic

NS

NS

* N

S

NS

NS

S.e.

m. 2

.16

NS

NS

NS

NS

NS

**

S.e.

m. 4

.74

* N

S N

S N

S N

S

**

S.e.

m. 3

.89

NS

**

NS

NS

**

***

S.e.

m.

1.07

aThe

leve

ls o

f ad

ded

Cu

corr

espo

ndin

g to

the

leve

ls o

f ad

ded

sulp

hate

are

0, 1

50, 3

00, 4

50, 6

00 a

nd 7

50 m

g C

u kg

-1.

NS

=N

ot s

igni

fica

nt; *

Pt0

.05

, **

PiO

.01,

*** P

<0.

001.

V

alue

s ar

e th

e m

ean

of f

our

obse

rvat

ions

.

Tab

le 4

. Mea

n liv

er li

pid

conc

entr

atio

ns, S

erum

cop

per l

evel

s, li

ver c

oppe

r co

ncen

trat

ion

and

tota

l liv

er c

oppe

r con

tent

of l

ayin

g he

ns g

iven

HzS

O4-

, C

uSO

4- a

nd m

ethi

onin

e-

supp

lem

ente

d di

ets

for 4

0 w

eeks

Leve

l of

addi

tives

St

atis

tical

sig

nifi

canc

e of

effe

ct

Bre

ed

(g so

4 kg

-l)

Met

hion

ine

and

-

Form

of

Leve

l of

Form

of

addi

tive

x (g

kg-

9 -

addi

tive

0 0

.2

0.4

0.6

0.8

1 .

o 1.

5 R

espo

nse

Bre

ed

addi

tive

addi

tive

leve

l of

addi

tive

Live

r lip

id c

once

ntra

tion

(g kg

-1 D

M)

Ove

rall

NS

NS

NS

NS

Lin

ear

NS

NS

274

H2S

04

242

285

320

306

270

200

Shav

er

cuso

4

319

240

275

298

186

Qua

drat

ic

239

War

ren

HzS

04

254

247

254

28 1

26

8 22

0 St

udle

r C

uSO

4 23

8 31

1 21

8 23

7 26

1

H2S

04

cuso

4 Sh

aver

War

ren

HzS

04

Stud

ler

CuS

O4

HzS

04

Shav

er

cuso

4

HzS

04

War

ren

Stud

ler

cuso

4

H zs

o 4

Shav

er

cuso

4

HzS

04

War

ren

Stud

ler

cuso

4

Seru

m C

u (p

g Zi

tre-')

30

0 28

3 25

8 24

3 22

0 26

3 24

0 28

3 28

3 22

5 20

3 35

5 29

3 28

8 31

0 28

8 28

5 31

3 30

8 29

3 25

3 24

5

Live

r Cu c

once

ntra

tion

(pg g-'

DM

)*

13.8

13

.3

11.4

13

.5

12.1

15

.4

(1.1

39)

(1.1

23)

(1.0

58)

(1.1

29)

(1.0

81)

(1.1

86)

11.9

18

.3

35.4

39

.1

209.

0 (1

,070

) (1

.262

) (1

.549

) (1

.592

) (2

.320

) 11

.6

11.0

16

.1

9.8

11.6

11

.2

(1.0

65)

(1.0

43)

(1.0

60)

(0.9

87)

(1.0

66)

(1.0

49)

11.7

11

.8

19.0

51

.5

41.6

(1

.068

) (1

.071

) (1

.278

) (1

.712

) (1

.619

)

Tota

l liv

er C

u (p

g)*

116

153

133

168

128

137

155

168

359

439

I442

133

156

120

138

127

127

140

157

174

475

353

(2.0

65)

(2.1

85)

(2.1

23)

(2.2

24)

(2.1

07)

(2.1

36)

(2.1

89)

(2.2

26)

(2.5

55)

(2.6

42)

(3.1

59)

(2.1

24)

(2.1

92)

(2.0

78)

(2.1

40)

(2.1

04)

(2.1

03)

(2.1

46)

(2.1

96)

(2.2

41)

(2.6

78)

(2.5

48)

280

325 14

.1

(1,1

50)

10.2

(I

,007

)

122

(2.0

85)

123 (2

.089

)

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

NS

NS

S.e.

m. 4

6.7

***

NS

***

NS

***

NS

NS

NS

S.e.

m. 2

1.8

***

***

***

***

***

***

***

NS

S.e.

m. (

0.07

85)

**

***

***

***

NS

S.e.

m. (

0,07

02)

***

***

***

~~

~

"The

leve

ls o

f ad

ded Cu c

orre

spon

ding

to

the

leve

ls o

f ad

ded

sulp

hate

are

0,

150,

300

, 450

, 600

and

750

mg

Cu

kg-l

. O

For

liver

cop

per a

naly

sis

of v

aria

nce

was

car

ried

out

usi

ng lo

g tr

ansf

orm

atio

ns a

nd th

e m

ean

valu

es a

re th

e an

tilog

s of

the

mea

n of

the

log

tran

sfor

mat

ions

with

the

mea

ns

NS

=N

ot s

igni

fica

nt; *

* P<

O.O

1, **

* P<O

.O01

. V

alue

s ar

e th

e m

ean

of f

our

obse

rvat

ions

.

of th

e lo

g va

lues

giv

en i

n pa

rent

hese

s.

Tab

le 5

. M

ean

prop

ortio

ns o

f fa

tty a

cids

in th

e liv

er f

at o

f la

ying

hen

s gi

ven

H&

04-,

CuS

Os-

and

met

hion

ine-

supp

lem

ente

d di

ets

for

40 w

eeks

Leve

l of

addi

tivea

St

atis

tical

sign

ific

ance

of

effe

ct

Bre

ed

(g so

4 kg

-l)

Met

hion

ine

and

(g k

g-I)

Fo

rm o

f Le

vel o

f F

orm

of

addi

tive

x ad

ditiv

e 0

0.2

0.4

0.6

0.8

1 .o

1.5

Res

pons

e B

reed

ad

ditiv

e ad

ditiv

e le

vel o

f ad

ditiv

e

War

ren

HzS

04

Stud

ler

CuS

O4

HzS

04

Shav

er

cuso

4

War

ren

HzS

04

Stud

ler

CuS

O4

War

ren

HzS

04

Stud

ler

CuS

Os

HzS

Os

Shav

er

CuS

O4

War

ren

HzS

04

Stud

ler

CuS

O4

13.8

12

.3

13'4

13

.2

13.0

11

.8

14. I

12"

11.7

11

.2

41.0

42

.0

35'8

42

.7

39.3

42'9

42

.2

44.7

45

.2

41 .O

13.5

14

.7

18.'

11.2

13

.7

10.5

11

.3

12'2

11

.3

10.4

3.0

3.1

3.4

3.

3 3.

4 4.

6 5

.0

3.7

3.1

3.1

I8:O 12

.8

13.7

11

.2

14.3

18: I

40.8

41

.7

42.1

38

.6

18:2

14.3

13

.3

11.5

10

.9

20:4

3.4

3.9

4.8

5.6

13.1

12.1

12.6

18

.5

15.5

19

.2

13.1

13

.5

19.2

15

.3

34'9

41

.5

38'6

40

.5

40.9

40

.3

34.5

41

.6

36.5

35

.4

13.3

15

.1

16.2

10

.7

12.7

10

.5

13.6

11

.0

11.4

13

.2

4.3

4.3

3.3

4.9

5.0

7

.0

4.8

4.

4 6

.8

6.2

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

Ove

rall

Lin

ear

Qua

drat

ic

NS

NS

***

***

NS

S.e.

m. 1

.44

NS

NS

* **

NS

S.

e.m

. 3.1

8

***

* N

S N

S

NS

S.

e.m

. 1.

54

NS

NS

NS

* NS

S.

e.m

. 1.3

3

NS

N

S

NS

NS

NS

NS

NS

N

S "3

NS

N

S

NS

&T

he le

vels

of

adde

d C

u co

rres

pond

ing

to t

he le

vels

of

adde

d su

lpha

te a

re 0

, 15

0, 3

00, 4

50,

600

and

750

mg

Cu

kg-1

. N

S=

Not

sig

nifi

cant

; * P<0

.05,

** P

<O

.OI,

*** P

<0.

001.

V

alue

s ar

e th

e m

ean

of f

our o

bser

vatio

ns.

1054 N. Jackson and M. H. Stevenson

by dietary treatment the overall mean weights kg-1 body-weight ( f s.e.m.) being 5.37 g k 0.093 and 0.092 g rf: 0.0032, respectively.

Mean liver lipid concentrations, serum Cu and the concentrations and total Cu content of the livers are given in Table 4. Liver Cu concentrations and total liver Cu content were linearly increased by level of additive (both P<O.OOl). Differing linear responses were shown by liver Cu concentra- tion and total liver Cu content (both P<O.OOlj within the forms of additive, the significant effects being produced by the added CuSO4 rather than the added HzS04. Blood serum Cu was signifi- cantly reduced by level of additive, the response being linear (both P < O . O O l j.

Liver lipid concentration was unaffected by dietary treatment, the overall mean ( k s.e.m.j being 260k9.5 g kg-l DM. The mean proportions of the fatty acids in the liver fat are presented in Table 5 . The 18:2 fatty acid was significantly affected (P<O.O5) by the form of additive but the 18 : 0, 18 : 1 and 20: 4 acids were unaffected. The overall effect of level of additive on the 18 : 0 and 18 : 1 acids was significant (P< 0.001 and 0.05, respectively), the response being positively linear for the 18:O acid (P<O.OOl) and negatively linear for the 18:l acid (P<O.Ol). Although the overall effect of additive on the 20: 4 acid was not significant, there was a positive linear response ( P < 0.05).

4. Discussion and conclusions

The mortality results suggest that the Shaver 288 were more susceptible to the dietary treatments than the Warren Studler birds.

The lack of effect of added HzS04 on food intake, egg numbers, mean and total egg weight and food conversion efficiency is in contrast with the adverse effects of added C U S O ~ . ~ ~ - ~ ~ The lack of response of methionine addition on the production indices measured indicates that the control diet was adequate in the sulphur-containing amino acids. The detrimental effects produced by added CuSO4 appear to be due to the considerable reductions in food intake above 0.4 g added sulphate kg-l and the concomitant reduction in AME and CP intakes-an effect not observed when the sulphate was added as the acid. The intake of these nutrients fa!ls below the Agricultural Research Council29 requirements for maintenance and maximum production. The finding that added HzS04 had no effect on food conversion efFiciency is in agreement with the observations of Sasse and Baker.16

The lack of effect of added CuSO4 on liver and gizzard weights kg-l body-weight (Table 3) is contrary to previously reported resultsz1* 23-25 but the observations for kidney weight agree with previous 25 The highest level of added CuSO4 significantly ( P < 0.05) depressed the liver weight per unit body-weight of the Shaver 288 birds compared with the liver weight of the birds given the lowest level of added CuSO4.

Of the 96 hens examined, 1 1 were not laying during the week before slaughter and of these, seven were in the groups receiving the three highest levels of added CuSO4. The remaining four birds were dispersed among the other groups. The overall lack of effect on oviduct weight per unit body-weight (Table 3) is obviously related to the lack of effect on egg production. The low mean oviduct weight kg-l body-weight of the Shaver 288 birds receiving the highest level of sulphate as added H2S04 was due to the virtual cessation of laying by two birds in this group and the overall effect was not significant. Ovary weight per unit body-weight of the Shaver 288 birds receiving the highest level of sulphate as HzSO4 was significantly reduced ( P < 0.05), a finding which suggests that ovarian regression was more advanced than oviduct regression.

The lack of effect of added HzS04 on liver Cu concentrations and total liver Cu content was expected. In the case of the CuS04-treated groups, the high liver Cu values, especially at the highest level of CuSO4 addition, were to be anticipated in the light of previous results.21 23 The threshold value observed in this experiment is the same as that found previously.21

Blood serum Cu levels have shown contrasting results in different e ~ p e r i m e n t s ~ l - ~ ~ and a response similar to the linear decrease in serum Cu with added CuSO4 observed in this experiment has been reported before.21 In this case, the added HzS04 produced a response in serum Cu similar to that produced by added CuSO4 although there is no obvious explanation for this effect.

The lack of effect of added HzS04 on liver lipid concentrations is expected since egg production

Copper sulphate in laying hen diets 1055

was also unaffected by additive but when cognisance is taken of previous reports indicating reduced liver lipid concentrationsz1, 23 in hens given added CuSO4, the overall lack of response found here is surprising. However, the liver lipid concentration of the Shaver 288 birds given the highest level of added CuSO4 was significantly lower (P< 0.05) than that of birds given the lowest level of additive.

The effects on the fatty acid composition of the liver fat of hens given added CuSO4 are in agreement with previous results.21 There was a decrease in the 18: l fatty acid, an observation which is contrary to that found in the pig given added CuS04.30 Also, the effects on the liver fatty acids of the hens given the added HzS04 paralleled those for added CuSO4.

The lack of effect of added HzS04 on the body-weight and egg production of laying hens con- trasts with the reduction in these parameters caused by added CuSO4 and suggests that it is the Cu ion rather than the sulphate ion which is producing these adverse effects.

Acknowledgements

The authors would like to thank the trustees of the Agricultural Research Institute, Hillsborough for experimental facilities, Dr S. T. C . Weatherup and Dr D. J. Kilpatrick of the Biometrics Division for assistance with statistical analyses and Mrs R. Park, Mr W. Graham and Mr G. McC Kirk- patrick for technical assistance.

References

I .

7 -. 3.

4.

5.

6.

7.

8. 9.

10.

1 1 .

12.

13. 14.

15.

16.

17.

18.

19.

Hawbaker, J. A.; Speer, V. C.; Jones, J. D.; Hays, V. W.; Catron, D. V. Effect of copper sulphate and anti- biotics on growth rate, feed conversion and faecal flora of growing pigs. J. Anirn. Sei. 1959, 18, 1505-1506. Braude, R. Copper as a stimulant in pig feeding (Cuprum pro pecuniu). World Rev. Anim. Prod. 1967, 3, 69-8 I . Wacker, A . ; Heyl, W.; Biiechl, H.; Holthoff, H. J. Zum Wirkungsmechanismus der Antibiotica als Wuchs- stoffe bei Tieren 2. Mitteilung: Versuche mit inaktiviertem Penicillin und CuS04. Arzneirn. Forscli. 1956, 6, 7 12-7 14. Mehring, A. L.; Brumbaugh, J. H.; Sutherland, A. I.: Titus, H. W. The tolerance of growing chickens for dietary copper. Poult. Sci. 1960, 39, 713-719. Kienholz, E. W.; Chappell, W. R.; Meglen, R. R.; Rudolph, H. In Trace sirbstances in environmental kedth- V U I . A symposium (Hemphill, D. D., Ed.), University of Missouri, Columbia, 1974, pp. 373-377. Poupoulis, C.; Jensen, L. S. Effect of high dietary copper on gizzard integrity of the chick. Poult. Sci. 1976, 55, 113-121. Waihel, P. E.; Snetsinger, D. C.; Ball, R. A.; Sautter, J. H . Variation in tolerance of turkeys to dietary copper. Poult. Sci. 1964, 43, 504-506. Aldringer, S. M. The effect of high copper levels on turkey performance. Poult. Sci. 1966, 45, 1065-66. Bowen, T. E.; Sullivan, T. W. Influence of dietary cupric sulphate on the response of young turkeys to peni- cillin-streptomycin ( 1 :3). Poult. Sci. 1971, 50, 273-278. Gordon, R. S.; Sizer, I. W. Ability of sodium sulphate to stimulate growth of the chicken. Science 1955, 122, 1270-1271. Coates, M. E.; Harrison, G. F. The effect on chick growth of inactivated penicillin, mineral sulphates or furazolidone supplements. Er. J . Nutr. 1959, 13, 345-355. Ross, E. ; Harms, R. H. The response of chicks to sodium sulphate supplementation of a corn-soy diet. Poult. Sci. 1970,49, 1605-1610. Martin, W. G. Sulphate metabolism and taurine synthesis in the chick. Poult. Sci. 1972, 51, 608-612. Ross, E.; Damron, B. L.; Harms, R. H. The requirement for inorganic sulphate in the diet of chicks for optimum growth and feed efficiency. Poult. Sci. 1972, 51, 1606-1612. Soares Jr, J. H. Experiments on the requirement of inorganic sulphate by the chick. Poult. Sci. 1974, 53, 246-252. Sasse, C. E.: Baker, D. H. Factors affecting sulphate-sulphur utilisation by the young chick. Poult. Sci. 1974,

Bornstein, S . ; Plavnik, Y . The sparing action of inorganic sulphate on sulphur amino acids in practical broiler diets. Preliminary trials with young chicks. Er. Porrlf. Sci. 1977, 18, 19-31. Plavnik, Y.; Bornstein, S . The sparing action of inorganic sulphate on sulphur amino acids in practical broiler diets. The capacity of sodium sulphate to replace some of the supplementary methionine in diets of chicks up to 5 weeks of age. Br. Poult. Sci. 1977, 18, 33-46. Husbands, D. R. The effect of dietary copper on the composition of adipose tissue triglycerides in the broiler chicken. Br. Portll. Sci. 1972, 13, 201-205.

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1056 N. Jackson and M. H. Stevenson

20.

21.

22.

23.

24.

25.

26.

27.

28. 29.

30.

Poupoulis, C.; Jensen, L. S. Effect of high dietary copper on fatty acid composition of the chick. Podr. Sci.

Jackson, N.; Stevenson, M. H. A study of the effects of dietary added cupric oxide on the laying, domestic fowl and a comparison with the effects of hydrated copper sulphate. Br. J . Nutr. 1981, 45, 99-1 10. Jackson, N. The effect of dietary copper sulphate o n laying performance, nutrient intake and tissue copper and iron levels of the mature, laying, domestic fowl. Br. J . Nutr. 1977, 38, 93-100. Jackson, N.; Stevenson, M. H.; Kirkpatrick, G. McC. Effects of the protracted feeding of copper sulphate- supplemented diets to laying, domestic fowl on egg production and on specific tissues, with special reference to mineral content. Br. J. Nub. 1979, 42, 253-266. Stevenson, M. H.; Jackson, N. Effects of level of dietary copper sulphate and period of feeding on the laying, domestic fowl with special reference to tissue mineral content. Br. J . Nutr. 1980, 43, 205-215. Stevenson, M. H.; Jackson, N. Effects of withdrawal of copper sulphate from the diet of the mature, domestic fowl with special reference to production and tissue mineral content. Br. J. Nutr. 1980,43,551-559. Stevenson, M. H.; Sackson, N. An attempt to distinguish between the direct and indirect effects, in the laying, domestic fowl of added dietary copper sulphate. Br. J . Nutr. 1981, 46, 71-76. Ichida, T.; Nobuoka, M. Determination of serum copper with atomic absorption spectrophotometry. Clin. Chim. Acta 1969, 24, 299-303. Lewis, D. A. The micro-determination of inorganic and total sulphate in urine. Analyst 1962, 87, 566-568. Agricultural Research Council The Nutrient Requirements of Farm Livestock No. 1, Poultry, Her Majesty's Stationery Office, London, 1975. Elliott, J. I.; Bowland, J. P. Effects of dietary copper sulphate on the fatty acid composition of porcine depot fat. J. Anim. Sci. 1968, 21, 956-960.

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