Studies on the absorption of volatile fatty acids in calves : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Animal Nutrition at Massey University of Manawatu

private study only. The thesis may not be reproduced elsewhere without

the permission of the Author.

IN

_CALVES

A thesis presented in partial. fulfillment of th e requirements for the degree

of

Dootor of Philosophy in Animal Nutri tion at Massey Universi ty

of Manawa tu.

RA1ISA.Y HANNA

KHOURI

1 966

I woul.d like to express my thanks and appreoiation to :-

Prof8ssor D.S. Flux for his guidanoe and encouragement, and for the statistical analysis

of

the experimental

data;

Professor D.A. Ti tohen for his advioe and enlightening disoussions;

Dr.

_G.W.

Butler of the Plant Ohemistry Division, D.S.I.R., for assistanoe

in the

radio-isotope analyses;

Dr .

_e.s.w.

Reid of the Plant Chemistry Division, D.S.I.R., for his

maqy

helpful suggestions;

Mr.

I.P.M. McQueen and the farm staff for their cooperation

in

the

mana

gement of the experimental animals;

Mr. G.H.

B

urns

for the reproduotion

of

the tables

_and

figures;

Miss

_M.G.

Campbell and the library statf for their helpful oooperation;

Mr.

B. Barnett, Miss

I .

Coxhead.,

_Mr.

D . Dellow and Mr. M. Wilson for their

_teohnical

help;

l(y wife, Soumaya, for her

_moral

support and patience during the o

ours

e

of

this stud1',

_ani

for her diligenoe

_in

typing this Thesis.

TABLE OF CONTENTS

Chapter

INTRODUCTION

The Rumen in Adult Bovines The Rumen in Calves at Birth

Volatile Fa tty Aoid Absorption Studies in Calves Based on the Analysi s of Rumen Contents

Vola tile Fa tty Aoid Absorption Studies in Calves Based on the Analysis of Blood

SECTION I

VOLATILE

FATTY ACID ABSORPTION STUDIES IN OONSCIOUS CALVES

I. POST-PRANDIAL CHANGES IN THE

VOIATILE

FATrY ACID AND GLUCOSE

Page

1 2 4

5 6

CONCENTRATIONS OF PORTAL BLOOD IN CONSCIOUS CAL

VES 1 2

REVIEW OF LIT ERATURE

1 3

Vol atile Fatty Aoid Absorption Studies in Mature Consoious

Ruminan ts Based on the Analysis of Portal Blood

1 3

Vola tile Fa tty Aoid Absorption Studies in Young Consoious

Calves Based on the Analysis of Portal Blood

1 5

MATERIAlS AND _MEmoDS

1 5

Experimen tal Design

1 5

Surgical Methods

1 8

- Anaesthesia

1 8

- Surgioal prooedure for th e f1stulation of the

rumen 1 8

- Surgical prooedure for the ca the teriza tion of the portal vein

1 9

- Pos t-opera ti ve medioa tion

1 9

- Care of portal vein oa the

ters 1 9

- Introduotion of catheters into the jugular vein

20

Chemical Methods

20

- Blood volatile f atty acids

21

- Blood glucose

23

- Rumen liquor vola tile fa tty aoids 24

Ohapter Page

I. RESULTS

26

Ohanges in the Volatile Fatty Aoid Conoentration and pH of

Rumen Liquor Following Milk Feeding and Following Grazing

26

Ohanges in the Vola tile Fa tty Aoid. and Gluoose Conoentra tions

of Porta1. Blood Following Milk Feeding _and. Following Grazing

34-

Differenoes in the Glucose and Vola tile Fa tty Aoid

Concen tra tions of Portal _and Jugular Blood Following _Milk

Feeding and Following Grazing

39

DISOUSSION

40

Ohanges in the Vola tile Fa tty Aoid Conoen tra tion and pH of

Rumen Liquor Following Milk Feeding and Following Grazing

41

Ohanges in the Volatile Fatty Acid and Glucose Concentrations

of Portal Blood Following Milk Feeding and Following Grazing

44-

Differences in _the Glucose and Vola tile Fa tty Acid.

Concan tra tions of Portal and Jugular Blood Following Milk

Feeding and Following Grazing

47

II. GLUCOSE: A

SOURCE

OF STEAM VOLATILE FA'rrY AOIDS IN _THE BLOOD OF CALVES

REVIEW OF LITERATURE

EXPERIMENTAL

Experimen t I. A Hyperglyoaemia Induced by the Feeding of Skim Milk Supplemented with Lactose

Ma terials and Methods Results

- Ohanges in the volatile fatty acid ooncentration and pH of rumen liquor following the feeding of whole milk and.

following the feeding of skim milk supplemented with lactose - Changes in the glucose and volatile fatty acid concentrations

of portal blood following the feeding of whole

milk

ani

50 51 52

52 52 54

following the feeding of skim milk supplemented with lactose

56

Experiment II,

A

Hyperglyoaemia Induoed by the Intravenous

Injection of Glucose

57

Materials and Methods

57

��b �

- Changes in the glucose and volatile fatty acid concentrations of

�

ar blood following t he intravenous injection of

50 % (w/v) D-glucose ani following the intravenous injeotion

of

0.9 %

NaCl

61

Chapter

II. Experiment III. A Hyperglyoaemia Induced by the Intravenous

Page

Injeotion of' Adrenaline

62

Ma terials and Methods

62

Results

66

- Changes in the gluoose and volatile tat� aoid ooncentrations ot jugular blood following the intravenous injection of

adrenaline and following the intravenous injeotion of'

0.9 %

NaCl

66

mSCUSSION

67

Experiment I . A Hyperglycaemia Induced by the Feeding ot

Skim Milk Supplemented with Laotose

67

Experiment II. A HypergIyoaemia Induced by the Intravenous

Injeotion of Glucose

69

Experiment III. A Hyperglycaemia Induced by the Intravenous

Injeotion of' Adrenaline

69

General

71

SUMMARY

73

III. STtIDIES ON THE _ABSORPTI ON _AND METABOLISM OF PREFORMED VOLATILE FATTY ACIDS IN OONSCIOUS CALVES BY THE ANALYSIS

OF JOOULAR BUXD

75

REVIEW OF LITERATURE

77

Absorption of' Preformed Vola tile Fa tty Acids from the Rumen

of' Oonsoious Calves Studied by the Analysis of Jugular Blood.

77

Metabolism ot Pref'onned Volatile Fat� Acids in Consoious Calves Studied. by the Analysis of Jugular Blood 77

EXPERIMENTAL

80

Experimen tal Animals

80

Fistulation

of

t he Rumen

83

Oa th eteriza tion ot the Jugular Vein

83

Experiment IV. Absorption of' Preformed Volatile Fatty Aoids tran the Rumen of Oonsoious Oal ves Studied by the Analysis

of Jugular Blood

84-

Experiment V. Metabolism of' Preformed Volatile Fatty Aoids

in Oonsoious Oalves Studied. by the Analysis _of Jugular Blood

86

RESULTS

87

Fas ting Levels of Vola tile Fa tty Aoids in Jugular Blood

87

Ohapter

III. Experiment IV. Absorption of Prefonned Volatile Fatty Aoids from the Rumen of Consoious Calves Studied by the Analysis

Page

of Jugular Blood

91

- Ohanges in the volatile fatt,y acid ooncentration of rumen oontents following the introduction of pref'onned sal ts of

these acids into the rumen

91

- Changes in the vola tile fa tty acid c oncentration of jugular blood following the introduction of prefonned salts of' these

aoids in to the rumen

91

Experiment V. Metabolism of Prefonned Volatile Fatty Aoids in

Consoious Oalves Studied by the Analysis of Jugular Blood

95

- Ohanges in the volatile fatty acid ooncentration of jugular blood following the injection of preformed salts of' these

aoids into the jugular vein

95

Heal th and. Growth Ra te of the Oal ves

95

DISOUSS! ON

97

Fasting Level s of Volatile Fatty Acids in Jugular Blood

97

Experiment IV. Absorption of Preformed. Volatile Fatt,y Acids from the Rwnen of Consoious Calves Studied by the Analysis

of Jugular Blood.

98

Experiment V. Metabolism of Prefonned Volatile Fatty Ac ids

in Conscious Oalves Studied by the Analysis of Jugular Blood.

1 00

Heal th and Growth Rate of _the Calves

101

SUMMARY

103

SECTION II

VOLATILE FA'rl'Y ACID ABSORPTION STUDIES IN ANAESTHETIZED OALVES

IV. CHANGES IN THE VENO-ARTERIAL VOLATIIE FATTY ACID CONCENTRATION DIFFERENCE OF

RUMINAL

BrooD roLLOWING THE INTRODOOTION OF PREFORMED VOLATILE FATTY ACIDS INTO

THE

RUMEN OF ANAESTHETIZED

CALVES

105

REVIEW OF LITERATURE

Vola tile Fa tt,y Acid Absorption Studies in Mature Anaes thetized

Rtuinants

Vola tile Fa tty Acid Absorption Studies in Young Anaesthetized Ruminants

106

106

109

Chapter . Page

IV. MATERIAIB

AND MEnIODS

109

Experimental Animals 109

Experimental Design 109

SurgioaJ. Prepa�tion

of

the Oalves 1 12

Experimental Prooedure 114

Prepara tion o� the Buffered Vola tile Fa tiu" Aoid Solution 1 17

ChemioaJ. Methods 1 18

RESULTS

126

Absorption

of

Volatile Fatty Aoids fran the Rumen 126 Blood Flaw Through the Right Ruminal Vein Catheter 129

pH

of Rumen Contents 131

Rumen Oapaci

_ty

1 33

Growth Rate of the Oalves 133

DISCUSSION 133

Absorption of Vola tile Fa tty Aoids from the Rumen 134- Blood Flow Through the Right Ruminal Vein Catheter 136

pH of

Rumen Contents 137

Rumen Oapaci

ty

138

Growth Rate

of

the Oal ves 139

SUMMARY 140

V.

_mE

APPEARANCE OF AOETATE- 1-<J14 IN RtlMlm EFFLUENT

AND

OABOn}) BrooD

FOLLOWING THE

INTRODUCTION OF PREFORMED VOLATILE FATTY

ACIDS INTO mE RUMEN OF ANAESTHETIZED OALVES 14 1

REVIEW OF LITERATURE

14 1

MATERIALS .AND METHODS

142

Experimental Animals 142

Experimen tal Design 142

Experimental

Prooedure

142

RESULTS

145

DISOUSSION 147

SUMMARY 148

CONCLUSION 149

1 50

APPENDIX 160

LIST OF TABLES Chapter

I.

POST-PRANDIAL

CHANGES IN

THE VOLATILE FATTY ACID AND

GLUCOSE CONCENTRATIONS OF

PORTAL BLOOD

IN CONSCIOUS

CALVES

Table Page

1

Rearing and. surgical details of calves used in milk and.

pasture fee ding absorption experiments

16

2

The age, weight, duration of fasting, quantity of milk oonsuned and time spent on pasture in milk and pasture

feeding absorption expe riments

17

3

Blood volatile fatty acid determination repeatability experiment. Results of eight volatile fatty acid

analyses on the same sample of bovine blood

22 4

Blood vola tile fa tty acid determination addition

experiment. Results of eight volatile fatty acid analyses on bovine blood to which known quanti ties of

aoetic acid were added

22

5

Blood glucose detennination repeatability experiment.

Resul ts of ten glucose analyses on the same sample of

bovine blood

23

6

Blood glucose detennina tion addition experiment.

Resul ts of five glucose analyses on bovine blood

to

which known quantities of glucose were added

23 7

Rumen liquor volatile fatty acid determination

repeatability experiment. Results of ten volatile fa tty acid analyses on the same sample of bovine

rumen liquor 24

8

Rumen liquor volatile fatty acid determination addition experiment. Results of ten vola tile fa tty acid analyses on bovine rumen liquor to whioh known quanti ties of

volatile fatty acids were added

24

9

^Ohanges in the volatile fatty acid conoentration and pH

of rumen liquor following m ilk feeding

27 10

Changes in the volatile fatty acid concentration

and

pH

of rumen liquor following grazing

27

11

Changes in vola tile fa tty acid and. glucose ooncentra tions

of portal blood following milk feeding

31 12

Changes in vola tile fa tty acid and gluoose concen tra tiona

of portal blood following grazing

3 1

Ohapter Table

I. 1 3 Dif'ferenoes in glucose ooncentration of portal

_and.

jugular blood following milk feeding

14- Differences in glucose ooncen tra tion of portal and jugular blood following grazing

15 Dif'ferences in the vola tile fa tty acid concen

_tra

tion

of

portal and jugular blood following milk feeding 1 6 Dif'ferences in the volatile fatty acid conoentration

of portal

_and.

jugular blood following grazing

II. GLUCOSE:

A

SOURCE OF STEAM VOLATILE FATTY ACIDS IN THE BLOOD OF CALVES

Table

17 Changes

in

the volatile fatty acid concentration and

pH

of rumen liquor following the feeding of whole milk and following the feeding of skim milk supplemen ted wi th laotose

18 Changes in the gluco se and vola tile fa tty acid

conoentrations of portal blood following the feeding of whole milk and following the feeding of skim milk supplemented with lactose

19 Details concerning the breed, age, weight and age at weaning of the calves that were u sed in the intravenous glucose infusion experiments

20 Changes in the glucose and volatile fatty acid conoentrations of jugular blood following the intravenous injection of 50 % ( w/v) D-glucose

_and

following the intravenous injection of 0. 9 % NaOl 21 De tails conceming the breed, age, sex

_and

weigh t

of

the calves used

in

the adrenaline experiment 22 Changes in the glucose and vola tile fa tty acid

concen tra tiona

_of

jugular blood following the intravenous injeotion of adrenaline

23 Changes in the glucose and volatile fatty

acid

concen tra tiona

of

jugular blood following the intravenous injection of 0. 9 % NaCl

III. STUDIES

ON THE

ABOORPTION

_AND

METABOLISM OF PREFORMED VOLATILE FATTY ACIDS IN CONSC IOUS CALVES BY THE ANALYSIS

OF

_Jm.ULAR

BlOOD Table

Page

3 5 35 36 36

56

58

58

62

63

63

24

Traoe mineralized sal t mixture, Feed Trade Manual Edition No.6, 1957, published by National Miller Publioation Inc., 6 E McDonald Rd., Prospect Heights,

Ill .

�

Ohapter Table Page III. 25 Details ooncerning the growth rates of the oal ves

together with the haemoglobin ooncentrations and. the packed oell volume of their blood. 82

26 The ohemical oanposi tion of 5 lit of the vola tile

fatty acid solution introduced into the rumen 85 27 The oomposition of 1 lit of the volatile fatty acid

solution used in the volatile fatty aoid tolerance tests 87

28 The vola tile fa tty aoid ooncentra tion of jugular blood following

₂₄

hours

of

fasting off

milk

and.

4.8 hours off pas

ture

87

29 Ohanges in the vola tile fa t ty aoid ooncen tra tions of jugular blood and rumen liquor following the introduotion of a buffered test solution of volatile

fa tty acids into the rumen, together with the amoWlt of 5 N phosphorio aoid required to maintain the pH of the rumen between 6.00

and

6.20

88

30

Ohanges in the vola tile fa tty acid concentration of jugular blood following the injection of a 4

^M

solution containing 85 % acetio and. 15 % propionic

aoid into the jugular vein 92

IV. CHANGES IN

THE

VliNO-ARTERIAL VOLATILE FATTY ACID OONCENTRATION

mFFERENOE

OF

RUMINA!,

BWOD roLLOWING THE INTRODUOTION OF PREroRMED VOLATILE FATTY

_ACIDS

INTO

THE RUMEN

O F ANAESTHETIZED

CALVES

Table

31 The ages, birth weights and weight gains of the calves

used in the acute experiments 108

32 Time oourse

of

the sampling sohedule followed during the acute volatile fatty acid absorption

experiments 117

33 Composition of 25 lit of volatile fatty aoid

rumen infusion 118

34 Volatile fatty acid oonoentration of blood collected simul taneously from the right

TlIIlinal

v ein

and.

carotid artery 120

35 Volatile fatty aoid concentration of arterial carotid blood for different groups in different periods.

Samples taken 15 minutes after the introduction

of

test solutions into the rumen 121

36 Vola tile fa tty acid concen tra tion of arterial carotid blood for different groups in different periods.

S�les taken

₃₀

minutes after the introduction of

test solutions into the rumen 122

Chapter Table Page IV.

37

Veno-arterial volatile fatt,y acid oonoentration

dif'ferences of blood. oollected simultaneously fran

the _ruminal vein and carotid artery

123 38

Veno-artllrial v.ola tile fa tty aoid oonoentra tion

differences for different groups in different pericxls. Samples taken

1 5

minutes after the

introduotion o f test solutions into the runen

124- 39

Veno-arterial vola tile fa tty aoid ooncentra tion

differenoes for different groups in different periods. Samples taken

30

minutes after the

introduotion o f test solutions into the rumen

125 40

Blood flow rates through the right ruminal vein

oa theter during aoute volatile fa tty aoid absorption

experiments

127

41

_Mean blood flow rates through the right _rwninal

vein oatheter

128

42

Changes in the Iii o f rumen contents during the

aoute vo latile fatty aoid absorption experiments

130 43

Rumen oapaoity of experimental calves

132

V.

THE

APPEARANCE OF

ACETATE-1_014

IN RUMEN EFFLUENT AND

CAroTID

BlOOD

FOLT.A>WING

THE

INTRODUOTION OF

PREroRMED

VOLATILE FATTY ACIDS INTO THE RUMEN OF

ANAESnIETIZED CALVES

Table

44 ^{The age} ani body weight of the calves

142 45

Volatile fatty aoid concentration _and. radio-aotivity of

blocxl oolleoted simultaneously from the right _ruminal

vein and carotid artery

143

46

Blood flow rates through

the

right nmdnal vein catheter during the acute vola tile fatty aoid

absorption experiments _{in which} radio-aotive aoeta te

was introduced into the I\UIlen

146

APPENDIX

Table

47

Analysis of varianoe of the volatile fatt,y aoid

oonoentration of arterial carotid blood for different groups in dif't'erent periods. Samples taken

1 5

minutes after _the introduotion _of test solutions

into the rumen

1

₆₀

Table Page

48

^Analysis of variance of the vola tile fatty aoid concentration

of

arterial o arotid blood for different groups in dif'ferent periods. Samples taken

30

minutes after the introduotion of test solutions into the rumen

160

49

Analysis of variance of the veno-arterial volatile f atty aoid oonoentration differences for different groups in different periods. Samples taken

1 5

minutes after the introduction of test solutions into the ^rumen

161

50

Analysis of varianoe of the veno-arterial vola tile fa tty aoid oonoentra tion differenoes fo r di£ferent groups i n different periods. Samples taken

30 minutes after the introd.uotion of test s olutions

into the rumen

161

5 1

Analysis at ^varianoe

of

th e mean blood flow rates

through ^the right ruminal vein oa the ter

162

LIST OF

FIGURES

Ohap ter Page

I.

POS

T-PRANDIAL CHANGES IN

T HE VOLAT ILE FATTY AOID

AND

GLUCOSE CONCENTRAT IONS OF PORTAL BIDOD

IN

O

ONS

CI O

US

OALVES

Figure

1

Thrombus

e noap sula ting the porta l vei n

oa

the ter 25 2 Chan ges

in

the vola tile fatty a oid oonoe ntra tion

an d pH of

:rume

n liquor following milk feed ing 28 3 Ohange s in the vol a tile f a tty a oid. o onoen tr a tion

and

pH of rume n liquor f ollo wing grazing 29 4- Oha nges

in

the vola ti le fa tty a oid and gluc ose

oonce n

tra.

tions

of

po rta l b lood foll owi ng

milk

fee ding 32

5 Ohang es

in

the vola tile f a tty ac id ani gluo ose

ooncen trati ons of porta l b lood f ollowin g grazing 33 6 Dif'f'e rence s in gluco se

oonc

e ntr a tion of porta l and

jugu la r b lood f ollowi ng

milk

fee ding 37 7 Diff erence s in glucose conc entra tion of portal an d

ju gul ar b lood f ollowi ng grazing 37

8 Diffe renoe s in

the

vo lati le f atty aci d oonc entra tion

of

portal

and

jugul ar b lood foll owin g milk feedi ng 38 9 Diff erence s

in

the volatile f a tty aoid. oonc entration

of

porta l and jugul ar b lood following gr azing 38

II. GLUCOSE, A

SOURCE

OF STEAM VOLATILE FATTY A OIDS

IN

THE BIOOD O F

^CALVES

Figure

10 Ohan ge s

in

the volati le fatty a oid. oonoen tra tion

and. pH of rume n liqu or follow ing the feedin g of who le milk

11

Chan ge s

in

the vo latile fa tty ao id c onoentra tion

and pH

of

rume n liqu or following the feedi ng of skim milk supp leme nte d with lac to se

12 Chan ge s in th e gluoo se and vola tile fa tty a oid.

oonce ntra tions

of

porta l b lo od fo llowing the fee ding o f who le milk

53

53

55

Chapter Figure Page

II .

13 Ohanges in the glucose and volatile �att,y aoid concentrations o� portal blood �ollowing the

�eeding o�

_skim milk

supplemented with laotose 55 14 Ohanges in the glucOse and volatile �att,y aoid

conoentra tions o� jugular blood �ollowing the

intravenous injeotion

_of

50 % (w/v) D-glucose 59 15 Changes in the glucose and vola tile �a tty aoid

oonoen tra tiona of jugular blood �ollowing

the intravenous injeotion o� 0.9 % NaOl 60 16 Changes in the glucose and volatile �atty acid

conoentra tions

_of

jugular blood �ollowing the intravenous injection

_of

a

_drenal

ine

17 Changes in the gluoose

_and

vola tile �a tty acid conoentrations

_of

jugular blood �ollowing

the intravenous injeotion

of

0.9 % NaGl 65 18 The �ormation of aoetio aoid �ran aoetyl coenzyme

A

70

III. STUDIES ON THE ABSORPTION AND

METABOLISM OF

PREroRMED VOLATILE

FATTY

ACIDS IN CONSarOUS CALVES

BY mE

ANALYSIS OF ^JUGULAR BWOD

Figure

19 Changes in the volatile �atty acid oonoentrations

of

rumen contents and jugular blood �ollowing the introduction of pre�onned vola tile �a tty acids into the rumen o� the

Milk

Fed Oalves

20 Changes in the volatile �atty aoid concentrations

of

runen contents and jugular blood �ollowing the introduction of pre�onned volatile �atty acids into the rumen o� the Pasture Fed Calves

21 Changes in the vola tile �a tty acid ooncentra tion

of

jugular blood �ollawing the injeotion of pre-

�ormed volatUe �a tty acids into the jugular vein of the Milk Fed Oalves

89

90

93 22

_Chan

ges in the volatile �atty acid concentration

of

jugular blood �ollowing the injection

_of

pre­

�onned volatile �atty acids into the jugular

vein

o� the Pasture Fed Calves 94

23 The rumen

waJ.l. in

a 15-week-old m:i.l.k-ted

oalt

102

24 The I'lIIlen wall in a 15-week-old pas

_ture-

reared

calf

102

Cha pter Page

IV.

CHANGES

IN

THE VENo-ARTERIAL

VOIATIIE FATTY ACID oaiOENTRATIOmr

DIFFERENCE OF

RUMINAL BLOOD FOLLOWING _THE

IN TRODUCTION OF PREroRMED VOLA TILE FATTY ACIDS I

NTO _THE

RUMEN OF ANAESTHETIZED

CALVES

Figur e

25 Entry into th e abdanina l oavi ty 26 Rum en oannula in posi tion

27 Cath eteri zation of mesen terio vei n

radioal

28 The righ t

ruminal

vein

29

An

ab sorp tion exp eriment in progress

30 Sohema. ti o diagram of the experimental pro oedu re used in th e aoute vo lati le fatty

acid

abso rption experiments

31 Ou tline of venous netw ork in the rumen wa ll f ollowing the inj eoti on of metqy lene b lue

32

Recordi ng of b lood pressure taken duri ng one of

the

ab sorpti on exp eriments

V.

_THE APPEARANCE ^OF

ACETATE- 1-014 IN RUMEN EFFLUEN T AND CAroTID

BIDOD FOLLOWING THE INTRODWTION

OF

PREFORMED VOLATILE

FATTY A CIDS

INTO THE

RUMEN OF

ANAESTHETIZED

CAL'Vm

Figu re

33 Radio-activity of rum en ef fluent and oarotid b lood samp les

1 10 110 1 1 1 111 1 13

1 15 1 16

1 19

144