Total cholesterol concentration in relation to superovulatory responses in crossbred cows

Tberiogenology

40:643-650,1993

TOTAL CHOLESTEROL CONCENTRATION IN RELATION TO SUPEROVULATORY RESPONSES IN CROSSBRED COWS 1

la M.Balakrishnan, V.K.Arora,

B.V.Bhaskar, 2 2 A. Ramu and

1 G.P.Chinna

iya, 2

P.A.Sarma

1 Station Embryo Technology Laboratory. Southern Regional Institute, Adugodi National Dairy Research Bangalore. 560 030. India 2 and Training Institute Central Frozen Semen Bank Regional Embryo Transfer Centre, Hesarghatta Bangalore. 560 088. India Received

for

pub1 ication: Accepted:

October 10, 1992 May 2, 1993

ABSTRACT

Blood serum total cholesterol levels of crossbred Taur-indicus donor cows (n=22), in their 1st to 4th parity, were studied as an indicator of embryo yield. These cows were superovulated either with FSH or PMSG + anti-PMSG on the 12th day of the synchronized estrous cycle. The total and transferable number of embryos did not differ significantly between the treatment groups. The number of corpora lutea and total and transferable embryos in donors having total cholesterol levels ~140 mg/dl were significantly (P140 mg/dl, indicating that low total cholesterol levels might adversely affect superovulatory response. Thus, estimation of total cholesterol concentrations of potential donors can be a useful tool for predicting superovulatory responses. Key

words:

blood cholesterol, superovulation

crossbred

cattle.

embryos.

Acknowledgements Director of the National Dai rv The authors thank the Research Institute of Karnal and the Head of the Southern Regional Station of Bangalore for providing the facilities; Professors J.Sitaramaswany and A.SubbaRao. for analysis; Dr.N.Pratap. M.Sreenath. statistical and L.Krishnamurthy, K.H.Rangappa. L.Louis. S.Prabhakaran for their excellent technical assistance. aCorrespondence

and

reprint

Copyright 0 1993 Butterworth-Heinemann

requests.

Theriogenology

644

INTRODUCTION Blood cholesterol plays a vital role in the phvsioloqv of reproduction. Blood cholesterol concentration and steroid synthesis are reported to have a positive relationship with energy/health status of an animal (1.2). There are reports indicating a positive correlation between hiqh blood cholesterol concentration and good reproductive performance in cattle (2-5). Furthermore, Japanese and German workers (6-9) have reported a close relationship between total cholesterol levels and superovulatorv response in cattle. It would be greatlv benefit embrvo transfer programs if the superovulatorv responses of potential donors could be before they are assiqned to superovulatorv predicted reqimens. Information on blood total cholesterol concentration in relation to superovulatorv responses in The ob.iective of our study was crossbred cattle is scarce. thus to estimate blood serum total cholesterol levels before superovulation and to establish its relationship to ovarian responses in crossbred cattle. MATERIALS

AND

METHODS

Twentv-two Taur-indicus (Holstein x Zebu: 3:l) cows. in their 1st to 4th lactation, were randomly allotted to receive 1 of 2 superovulation treatments. On the 12th dav of Gro p I and Group II donors the synchronized estrous cvcle. were superovulated with either FSH; ! (42 mg; NIH standard pituitarv). in descending doses or a single intramuscular injection of PMSG’ (2500 to 3000 IU). respectively. The total FSH dose was divided into 8 in.iections (8/8. 6/6. 4/4 i.m) and were given twice daily at 12 hours and 3/3 mq. intervals for 4 consecutive days. At 48 hours after the first qonadotropin in.iection. the 30~s were administered a i.m twice at 12 prostaglandin analoque. Luprostiol 15 mq. hours intervae) . All donors were administered 2.5 ml of 6 hours GnRH anal oque (Buserelin acetate 0.01 mq. i.m) before AI. The donors were inseminated at 12. 18. and 30 hours after the onset of estrus using frozen - thawed semen. Just before AI, Group II cows w re qiven an intravenous ? (5 ml ). On Day 7 after the injection of monoclonal anti-PMSG first AI. embrvos were recovered nonsurgicallv after number of corpora lutea and assessment of ovarian size. unovulated follicles, palpation per rectum. The embrvos were examined both morpholoqicallv and qualitativelv under a stereomicroscope (10). Jugular blood samples (5 ml) were co1 lected on the last 3 consecutive davs before supercholesterol levels were ovulation, and the serum total

Inc.. London, Canada. b Folltropin Vetrepharm c FolligonIntervet International. B.V.. Boxmeer. Holland. d ProsolvinIntervet International. B.V.. Boxmeer. Holland. e Receptal - Hoechst, Veterinar GmbH. Munchen, Germany. f Neutra PMSG - Intervet International.B.V.,Boxmeer.Holland.

Theriogenology

Table

1.

645 Means +SEM of and transferable cows having concentrations

Groups

No.of cows

unovulated number of low and high

foll icles and total embryos in crossbred total cholesterol

CL,

CL

Unovul ated follicles

No. Total

-.___.._ Low cholesterol (t140 mq/dl)

0 to

Range High cholesterol (>140 mq/dl )

1.9kl.l

11

11.6+1.1b

R

4

Embryos

Transferable __.-_-

_-..___.__-__ 11

of

n 9.2+0.8>

0.8+1

6 to

0

15

2.2+0.8”

to

11.2g.2b

.2n

4

0.320.7

0 to

2

7.4kO.l

b

5to19 oto4 6 to 20 Range 2 to 15 .-_________ - ~._._ __I-. __. ____-.-_-a.b Means within a column with different superscripts differ significantly (P
unovulated follicles. The relationship of total cholesterol concentration (on the day of flushing) on the number of CL. unovulated foll icles and total and transferable number of embryos were tested with simple linear regression. The fitted regression 1 ines of the ovarian responses to total cholesterol are depicted in Figure 1. It is evident. as the total cholesterol level rises the superovulatory responses increases in terms of number of CL and the total and embrYes at the rate of 0.13. 0.15 transferable number of and 0.11. respectivelv. per unit rise in total cholesterol unovulated follicles is found (Figure 1 B,C and D). However. to decrease with an increase in total cholesterol level at the rate of of 0.11 per unit rise in total cholest,erol level (Figure 1 A). Furthermore. a multiple linear regression equation was fitted to predict the total number of recoverable embrvos based on total cholesterol concentration. the number of unovulatorv foll icles and CL: TER = -7.463+0.045 CH0+1.003 CL+O.229 UF (Y q a+blxl+b?,2+b3,3 ). It can be observed that the most significant parameter contributing to the total number of recoverable embrvos is the CL. provided the total cholesterol levels are sufficiently high in the donors. Other factors remaininq constant. the per unit increase in the CL is the recovery of 1 additional embrvo.

646

Theriogenology

estimated soectroohotometricallv animal. the mean of the 3 for statistical analvsis.

dav

usinq values

readv kits. (CV 0.65%)

gFor was

each taken

For testlnq the differences between the 2 treatment qr-ouos. ANOVA procedure was carried out. The unclassified data on the total cholesterol levels of the donors (irresoective of treatment) were subiected to frequencv distribution with uniform class intervals. The median was calculated and taken as the reference point for classifvinq the donors into low (cl40 mq/dl) and hish (>140 mq/dl) total cholesterol rlrouos. Then the data on low (n=ll) and hiqh (n=ll ) total cholesterol qrouos were analvsed statisticallv (11) for ANOVA. correlation coefficients and simple and multiole reqressions. The mean values were exoressed with the standard error of the means. RESULTS

AND

DISCUSSION

The total and transferable number of embrvos in Group I (6.2+2.0 and 5.8+1.8. resoectivelv: n=ll) and in GrOUD II (4.1+1 .O and 4.6+1 .5. resoectlvelv: n=ll ). did not differ siqnificantlv between the 2 treatment reaimens. which was significant to that of earlier reoorts (6.7) but not to that of Dielman (12). The mean concentration of total cholesterol levels before suoerovulation ranged between 79 and 182 mq/dl. In the low level qrouo (<140 ms/dl) and the hicih level (>140 mq/dl) qrouo total cholesterol ranged between 79.5 and 133.5 and 141.5 and 182 mq/dl. resoectivelv. The low total cholesterol qrouo (n=ll) vielded 9 embrvos, of which onlv 3 in the hiah ! 33.3%) were of transferable aual itv. whereas: concentration qrouo (n=ll) 83 Of 123 embrvos (67.5%) were of transferable aualitv. total and The averase number of CL and the transferable number of embrvos were sinnificantlv lower (PCO.5) in donors havinq <140 mq/dl of total cholesterol than in cows havinq hish (>140 mq/dl) concentrations (Table The low total cholesterol qrouo showed a sianificantlv 1). hisher (P
Diasnostic

Svstem$.

Ethnor

Ltd..

Bombav.

India.

647

Theriogenology

#

15

1

-

l.JF=20.30-0.11

12

T-CHO (A)

(B)

16 12 8 4 0 12 i

70

TE =10.17+0.11

90 TOTAL

Figure

1.

110

T-CHO

130

150

CHOLESTEROL

170 (mg/dl)

I

190

Regressions of unovulated follicles (A). total embryos recovered Corpora lutea (B). (Cl and transferable embrvos (D) on total cholesterol concentration in crossbred cows (Data were derived from counts made on Dav 7. after first A.1).

Theriogenology

646

A high concentration of total cholesterol and high reproductive performances were reported in cattle fed a 120% NRC dietarv enerqy level (21. Positive energy balance has been associated with high circulating concencentrations of progesterone (P4) during the luteal phase before ( 13 1 and after AI (14). Cholesterol is a precursor for luteal cell P4 and can be derived from cellular de novo synthesis. svnthesis or bv the uptake of plasma lipoprotein cholesterol. In-vitro studies indicate that steroidoqenesis bv luteal tissue. is dependent on the provision of lipoprotein cholesterol (151. The significant relationship cholesterol between total and plasma P4 has been demonstrated in Holstein heifers fed diets containing 15% whole sun flower (161 and in suckled Brahman crossbred cows fed whole cotton seed (17). However, evidence to support a cause and effect relationship between elevated total cholesterol and P4 is limited (181. A low total cholesterol concentration might indicate the health/ener9y status of the which in turn would affect the follicular and oocvte donors, resulting in poor ovulation and embrvo vield. A maturation, faulty metabolism (9). or phvsiolow enerqy diet (191. pathological stress /4.5.20.211. fattv liver or fat cow 231 have al 1 been suggested to be causative syndrome (22. factors for low total cholesterol concentration in animals. The large number of unovulated follicles in the low total cholesterol group suqgests that follicular development did occur in response to the superovulatorv treatments. but foll icles were sub-normal. perhaps the function of those such that the LH surqe was not elicited properly and thev the role of total cholesterol in failed to ovulate. However. needs detailed biochemical the reproduction of cattle. studies involving fat supplementation in relation to plasma cholesterol and progesterone concentrations in dairv with particular reference to superovulatorv animals. responses.

In conclusion, the present studv confirms earl ier reports (6-9) that low total cholesterol levels mav affect The total cholesterol estimation is superovulatorv response. simple and does not require expensive equipment. Thus, the donors based on total preliminary selection of potential is suggested for use in cholesterol concentrations. predicting ovarian response after superovulation.

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