The Relationship between Breed and Litter Size in Sheep and Maternal Serum Concentrations of Placental Lactogen, Estradiol

and Progesterone W. R. Butler, S. M. Fullenkamp, L. A. Cappiello and S. Handwerger J ANIM SCI 1981, 53:1077-1081.

The online version of this article, along with updated information and services, is located on the World Wide Web at: http://www.journalofanimalscience.org/content/53/4/1077

www.asas.org

T H E R E L A T I O N S H I P B E T W E E N B R E E D A N D L I T T E R S I Z E IN SHEEP A N D M A T E R N A L S E R U M C O N C E N T R A T I O N S OF PLACENTAL LACTOGEN, ESTRADIOL AND PROGESTERONE 1

W. R. Butler 2, S. M. Fullenkamp, L. A. Cappiello and S. Handwerger a
Cornell University, Itbaca N Y 1 4 8 5 3

Summary Milk production in sheep is known to be affected by breed and litter size. To investigate a possible hormonal basis for such effects, we measured maternal serum concentrations of placental lactogen, estradiol and progesterone during late gestation in sheep of several breeds and ages. Circulating concentrations of placental lactogen were directly related to litter size in all breeds, with the greatest concentrations being observed in Finnish Landrace ewes bearing three lambs. Among monotocous animals, no significant breed differences were observed, but ewe lambs bearing crossbred fetuses had higher placental lactogen concentrations than ewes of the same breed carrying purebred offspring. Placental lactogen concentrations were similar in all animals bearing twins, except for Corriedale ewes, which had the lowest concentrations. Serum estradiol concentrations 2 to 5 weeks prepartum did not differ between ewes of different breeds or ewes with different litter sizes. Progesterone, like placental lactogen, was related to litter size and, presumably placental mass. The relationship between litter size and placental lactogen concentrations, together with the absence of difference in estradiol secretion, suggests that differences in production of the lactogenic hormone may contribute to the superior lactational performance that has been reported for ewes which bear muhiple offspring. (Key Words: Placental Lactogen, Estradiol, Progesterone, Sheep.)

Introduction
The identification and measurement of placental lactogen (PL) during gestation in domestic ruminants (see Blank et al., 1977, for review) has led to studies showing a high correlation between circulating levels of this hormone and postpartum milk production by c a t d e (Bolander et al., 1976) and goats (Hayden et al., 1979). Furthermore, PL production in goats is directly related to the number of young in utero, thus providing a mechanism by which the mammary gland is suitably prepared for future demands (Hayden et al., 1979). It is known that the number of offspring influences milk production in sheep (Alexander and Davies, 1959; Davies, 1963; Gardner and Hogue, 1964; Peart, 1967; Peart et al. (1972, 1975b), and, in addition, breed differences in lactational ability have been reported (Gardner and Hogue, 1966). The purpose of the present study was to investigate the effect of breed and litter size on the secretion of PL and other mammogenic hormones in sheep.

Materialsand Methods

Blood samples were collected by jugular puncture biweekly from mating to parturition from nine Dorset ewe lambs. Blood was collected biweekly beginning 6 to 8 weeks before expected parturition from ewe lambs and mature ewes of the Hampshire, Corriedale and Finnish Landrace (Finnsheep) breeds. All animals were housed in adjacent pens and were fed a diet of grass silage, hay and ground concentrate. Each group of ewes was mated to a single ram. Hampshire and Corriedale ewe lambs were mated to Suffolk 1The authors express appreciation to Dr. P. A. rams, while other ewes were mated to a purebred Ohenacu for statistical analysis, R. Karaka, F. Michel ram of their own breed. All lambings occurred and V. Nicoll for technical assistance and to L. Taylor between January 1 and April 15, 1978, and, for assistance in typing this manuscript. within a breed-age group, parturition dates 2 Dept. of Anita. Sci. Duke Univ. Medical Center, Durham, NC 27710. ranged over no more than 3 weeks. 1077 JOURNAL OF ANIMAL SCIENCE, Vol. 53, No. 4, 1981

1078

B U T L E R ET AL.

Blood samples were allowed to clot and then were refrigerated (4 C) for 24 hr before centrifugation. Serum samples were stored at - 2 0 C until aliquots were removed for radioimmunoassay of PL (Handwerger et al., 1977), progesterone (Gengenbach et al., 1978) and estradiol. Estradiol-173 was measured with antibody GDN-#244, which is similar to that previously described by Korenman et al. (1974). The antiserum was diluted to an initial concentration of 1:50,000 and (2,4,6,7,16,17-3H) estradiol 4 was used as tracer after Sephadex LH-20 purification. Serum samples (400 to 1,000/J1) in duplicate were extracted with 5 ml freshly redistilled benzene: toluene (2/1 v/v). Estradiol from serum extracts was isolated on Sephadex LH-20 columns (50 mg) eluted with chlorofbrm: methanol: heptane: water (500:225:1500:9). Procedural losses were measured by the addition of 200 to 500 cpm (3H) estradiol before extraction (Chenault et al., 1975). Mean recovery averaged 84%. A similar extraction efficiency (75 to 90%) was calculated from the recovery of estradiol added to serum. Charcoaltreated serum from cows was used as the assay blank for the calculation of serum samples. This serum correction was .5 to 1.5 pg/ml. Pooled serum samples were included in each hormone assay at three volumes for determination of intra- and interassay coefficients of variation; for PL, estradiol and progesterone, the coefficients were 8 and 11% (n = 6), 10 and 12% (n = 8) and 5 and 7% (n = 8), respectively. Hormones concentrations in two samples from each ewe collected between 2 and 5 weeks prepartum were averaged and subjected to least-squares analysis of variance. Hormone concentrations were compared among breed-age groups within litter size by Duncan's new multiple range test.

concentrations occurred simultaneously with the increase in PL concentrations (Kelly et al., 1974). PL concentrations were maximal at week 19 which is in agreement with C h a n e t al. (1978) but somewhat later than the period when peaks were detected by a radioreceptor assay technique (Kelly et al., 1974, 1976). This difference may have been due to the techniques used or to variation among animals. Because our results for Dorset ewe lambs and all the reports cited above indicated that maximum PL concentrations occur during late gestation, blood samples were collected during this period for evaluation of breed differences. Mean serum PL concentrations between 2 and 5 weeks prepartum for several breed-age groups are shown in table 1. Analysis of variance showed highly significant differences among breeds and among number of offspring, but no significant breed x litter size interaction. The greatest differences in mean concentrations of PL were related to litter size, and concentrations increased with increasing number of offspring. This was true in all breed-age groups. PL concentrations were highest in Finnsheep bearing three lambs. Comparison of serum PL concentrations in Finnsheep showed no differences due to age or previous parity in the secretion of this hormone. This was not the case for monotocous Corriedales and Hampshires; the lambs of each breed had higher circulating concentrations of PL than did the adults. Lambs in these two groups also had higher PL concentrations than the

N-9 1,2~3

803

Results and Discussion

The pattern of serum concentrations of PL and progesterone during gestation in Dorset ewe lambs is shown in figure 1. PL first became detectable in the peripheral circulation at about 60 days of gestation, in accordance with previous reports (Kelly et aL, 1974, 1976; Handwerger et al., 1977; Robertson et al., 1980). An increase in serum progesterone

2C0

4 New England Nuclear, Boston, MA.

Figure 1. Maternal serum concentrations of placental lactogen (oPL) and progesterone (PROG) during gestation (weeks) in Dorset ewe lambs. Two of the animals bore twins.

PLACENTAL LACTOGEN AND LITTER SIZE TABLE 1. THE RELATIONSHIP BETWEEN BREED AND NUMBER OF OFFSPRING ON SERUM PLACENTAL LACTOGEN (PL) AND ESTRADIOL (E) CONCENTRATIONS IN PREGNANT LAMBS AND MATURE EWESa PL in ewes with varying number of offspring, ng/ml Breed-age group Finnsheep ewes Finnsheep lambs Corriedale ewes Corriedale lambs Hampshire ewes Hampshire lambs Dorset lambs 1 1,197 + 559(3) x 727 +- 150(4)Y 669 53(7) bc 1,260 + 163(10) b 939 137(8) 1,271 153(9) c 790 +- 205(7) 2 1,584 186(16) 1,672 + 421(6) 1,267 167(12) d 1,831 246(6) d 1,895 + 428(3) 1,343 706(2) 3 2,997 289(6) x 2,802 665(4)Y E, pg/ml 9.9 10.2 11.3 9.4 12.8 8.6

1079

.9(25) 1.8(14) 1.5(19) .7(10) 1.3(14) 1.3(12)

avalues listed are mean + standard errors; numbers in parentheses are numbers of animals. b'c'd~X'YMeanconcentrations within rows and columns with the same superscript are different (P<.05).

other lambs (Dorset and Finnsheep), but this difference was not statistically significant. The enhanced hormone production in the Corriedale and Hampshire lambs may have been an effect of crossbreeding, as they were mated to Suffolk rams, whereas all other animals were bearing purebred lambs. More direct tests of the effect of crossbreeding, as well as more detaiJed information on the secretion of PL in the Suffolk breed, are needed. Among ewes bearing twins, the only significant difference in PL concentrations was between Hampshire and Corriedale ewes (table 1). Estrogen and progesterone are important components in the mammogenic complex in a number of species (Anderson, 1974). They were measured in the present study for an assessment of breed and litter size effects. Serum estradiol concentrations were remarkably consistent within litter sizes, and the data were pooled within breed-age groups (table 1). Thimonier e t al. (1977) have reported that total estrogen concentrations on day 102 of pregnancy are related to numbers of lambs born and total birth weight. Thus, although estradiol secretion may serve important functions such as inducing the onset of labor (Currie, 1977), total serum estrogens seem to have a better correlation with fetal-placental mass. Serum progesterone concentrations during late gestation increased as number of fetuses increased (table 2), in suport of previous observations (Gadsby e t al., 1972). Progesterone concentrations were generally lower in Finnsheep than in Hampshire or Corriedale ewes.

Although birth weights of lambs were not available in all groups, Finnsheep lambs weighed less than Hampshire or Corriedale lambs (2.7 + .1 kg, n = 54;4.3 +.3 kg, n-- 9, a n d 4 . 5 + . 2 kg, n = 20, respectively). The positive relationships between weight and number of lambs and placental mass (Alexander, 1964) are the likely explanation for the observed differences in progesterone secretion, because this hormone derives primarily from the placenta during late gestation (Linzell and Heap, 1968). Milk production in both sheep (Alexander and Davies, 1959; Davies, 1963; Gardner and Hogue, 1964; Peart, 1967; Peart e t a l , 5972, 1975b) and goats (Hayden e t al., 1979) is directly related to the number of offspring. Enhanced milk secretion following the birth of multiple young is apparent within the first week postpartum in both species. The effects of multiple births on milk yield have been attributed to increased suckling intensity (Alexander and Davies, 1959; Davies, 1963). Milk production was 150% greater in ewes bearing and nursing twins than in ewes either bearing singles or ewes bearing twins, but nursing a single lamb (Alexander and Davies, 1959). Unfortunately, a group of ewes that gave birth to a single lamb but nursed two lambs was not included. When a study was conducted in which this nursing regimen was investigated, milk production was n o t increased by the greater suckling intensity (Moore, 1966). These results suggest that greater potential for milk yield in multiparous ewes is largely determined before parturition and that realization of such potential is modified

1080

BUTLER ET AL. TABLE 2. SERUM PROGESTERONE CONCENTRATIONS(NANOGRAMS/M1LL1LITER) DURING LATE GESTATION IN MATURE EWESa No. of offspring

Breed Hampshire Corriedale Finnsheep

1 6.53 .83(8) b 4.06 -+. 32(7)bd 4.45 .24(3)

2 8.54 -+.98(6) f 7.10 .44(12) de 4.75 .23(16) cef

5.81 +-.46(6) c

avalues listed are mean standard errors; numbers in parentheses are numbers of animals. b'c;d'e'fMeans within rows and columns with the same superscript differ (b,c P<.05; d,e,f P<.O01).

by the suckling activity of the lamb(s). This is consistent with the direct effect of multiple offspring on mammary growth during late pregnancy in ewes (Rattray et al., 1974) and with the observation that ewes that gave birth to twins but were suckled by only one lamb produced more colostral milk than ewes bearing and suckling single lambs (Peart et aL, 1972). In light of the mammogenic activity of PL (Martal and Djiane, 1976), the positive association between PL concentration and number of fetuses in sheep (present data; Handwerger et al., 1977; Thimonier et al., 1977; Gluckman et al., 1979) and goats (Currie et al., 1977; Hayden et al., 1979) provides an explanation for the previously observed relationship between milk yield and number of offspring in these species. Genetic differences in milk production by ewes have been reported (Gardner and Hogue, 1966; Peart et al., 1979), which may be partly related to genetic differences in PL secretion. Lactational performance of Hampshire ewes was superior to that of Corriedale ewes (Gardner and Hogue, 1966), which is consistent with the difference in PL levels between these breeds. The milk production by ewes bearing crossbred lambs was greater within the first week of lactation than that by similar ewes bearing purebred lambs (Peart et al., 1975a), a finding consistent with enhanced PL concentrations in lambs bearing crossbred offspring. Genetic differences in suckling activity are also involved (Moore, 1966). In order to maximize milk production for increased efficiency of lamb production, further studies should be conducted to characterize genetic (breed or sire) effects on PL secretion and the relationship of PL to subsequent mammary function. A synergistic relationship hetween PL and estrogen on mammary development might be

expected from the accepted actions of these hormones. Total estrogen concentrations on day 110 of pregnancy in ewes were positively correlated to milk production and growth of Iambs (Terqui and Delouis, 1975), which is consistent with the positive effects on lactation of estradiol given to induce parturition (Bosc et al., 1977). Although estradiol and PL may synergistically stimulate mammary development, the uniformity of estradiol concentrations in all breed-age groups studied suggests that differences in milk production with varying number of fetuses are more likely attributable to the differences in serum PL concentrations. However, major changes in estradiol secretion occur just before parturition (Currie, 1977), and differences due to fetal number could be expressed at that time and would not be observed under the sampling schedule used in the present study.
Literature Cited

Alexander, G. 1964. Studies on the placenta of the sheep (Ovis aries L.): Placental size. J. Reprod. Fertil. 7:289. Alexander, G. and H. L. Davies. 1959. Relationship of milk production to number of lambs born or suckled. Australian J. Agr. Res. 10~720. Anderson, R. R. 1974. Endocrinological control. P. 94-147 In B. L. Larson and V. R. Smith (Ed.) Lactation: A Comprehensive Treatise. Vol. I. Academic Press, New York. Blank, M. S., J.S.D. Chan and H. G. Friesem 1977. Placental lactogens, new developments. J. Steroid Biochem. 8:403. Bolander, F. F., Jr., L. C. Ulberg and R. E. Fellows. 1976. Circulating placental lactogen levels in dairy and beef cattle. Endocrinology 99:1273. Bosc, M. j., C. Delouis and M. Terqui. 1977. Control of the time of parturition of sheep and goat. In Management of Reproduction in Sheep adn Goats Symposium. Univ. of Wisconsin, Madison. Chan, J.S.D., H. A. Robertson and H. G. Friesen.

PLACENTAL LACTOGEN AND LITTER SIZE 1978. Maternal and fetal concentrations of ovine placental lactogen measured by radioimmunoassay. Endocrinology 102:1606. Chenault, J. R., W. W. Thatcher, P. S. Kalra, R. M. Abrams and C. J. Wilcox. 1975. Transitory changes in plasma progestins, estradiol and luteinizing hormone approaching ovulation in the bovine. J. Dairy Sci. 58:709. Cuttle, W. B. 1977. Endocrinology of pregnancy and parturition in sheep and goats. In Management of Reproduction in Sheep and Goats Symposium. Univ. of Wisconsin, Madison. Currie, W. B., P. A. Kelly, H. G. Friesen and G. D. Thorburn. 1977. Caprine placental lactogen: Levels o f prolactin-like and growth hormone-like activities in the circulation of pregnant goats determined by radioreceptor assays. J. En docrinol. 73:215. Davies, H. L. 1963. The milk production of Merino ewes at pasture. Australian J. Agr. Res. 14:824. Gadsby, J. E., R. B. Heap, D. G. Powell and D. E. Waiters. 1972. Diagnosis of pregnancy and the number of fetuses in sheep from plasma progesterone concentrations. Vet. Rec. 90:339. Gardner, R. W. and D. E. Hogue. 1964. Effects of energy intake and number of lambs suckled on milk yield, milk composition, and energetic efficiency of lactating ewes. J. Anita. Sci. 23 : 935. Gardner, R. W. and D. E. Hogue. 1966. Milk production, milk composition and energetic efficiency of Hampshire and Corriedale ewes fed to maintain body weight. J. Anim. Sci. 25:789. Gengenbach, D. R., N. Butendieck, P. M. Riek, R. L. Scipioni, E. B. Oltenacu and R. H. Foote. 1978. Controlled superovulation in dairy heifers using prostaglandin F2a and pregnant mare serum gonadotropin. J. Anita. Sci. 46:1293. Gluckman, P. D., S. L. Kaplan, A. M. Rudolph and M. M. Grumbach. 1979. Hormone ontogeny in the ovine fetus. II. Ovine chorionic somatomammotrophin in mid- and late gestation in the fetal and maternal circulations. Endocrinology 104:1828. Handwerger, S., C. Crenshaw, Jr., W. F. Maurer, J. Barrett, T. W. Hurley, A. Bolander and R. E. Fellows. 1977. Studies on ovine placental lactogen secretion by homologous radioimmunoassay. J. Endocrinol. 72:27. Hayden, T. J., C. R. Thomas and I. A. Forsyth. 1979. Effect of number of young born (litter size) on milk yield of goats: Role for placental lactogen. J. Dairy Sci. 62:53. Kelly, P. A., H. A. Robertson and H. G. Friesen. 1974. Temporal pattern of placental lactogen and progesterone secretion in sheep. Nature (London) 248:435. Kelly, P. A., T. Tsushima, R.P.C. Shiu and H. G.

1081

Friesen. 1976. Lactogenic and growth hormonelike activities in pregnancy determined by radioreceptor assays. Endocrinology 99: 765. Korenman, S. G., R. H. Stevens. L. A. Carpenter, M. Robb, G. D. Niswender and B. M. Sherman. 1974. Estradiol radioimmunoassay without chromatography: Procedure, validation and normal values. J. Clin. Endocrinol. Metab. 38:718. Linzell, J. L. and R. B. Heap. 1968. A comparison of progesterone metabolism in the pregnant sheep and goat: Sources of production and estimation of uptake by some target organs. J. Endocrinol. 41:433. Martal, J. and J. Djiane. 1976. Mammotrophic and growth promoting activities of a placental hormone in sheep. J. Steroid Biochem. 8:415. Moore, R. W. 1966. Genetic factors affecting the milk intake of lambs. Australian J. Agr. Res. 17:191. Peart, J. N. 1967. The effect of different levels of nutrition during late pregnancy on the subsequent milk production of Blackface ewes and on the growth o f their lambs. J. Agr. Sci. (Camb.) 68:365. Peart, J. N., J. M. Doney and A. J. MacDonald. 1975a. The influence of lamb genotype on the milk production of blackface ewes. J. Agr. Sci. (Camb.) 84:313. Peart. J. N., J. M. Doney and W. F. Smith. 1979. Lactation pattern in Scottish Blackface and East Friesland Scottish Blackface crossbred ewes. J. Agr. Sci. (Camb.) 92:133. Peart, J. N., R. A. Edwards and E. Donaldson. 1972. The yield and composition of the milk of Finnish Landrace Blackface ewes. I. Ewes and lambs maintained indoors. J. Agr. Sci. (Camb.) 79:303. Peart, J. N., R. A. Edwards, and E. Donaldson. 1975b. The yield and composition of the milk o f Finnish Landrace Blackface ewes. II. Ewes and lambs grazed on pasture. J. Agr. Sci. (Camb.) 85:315. Rattray, P. V., W. N. Garrett, N. E. East and N. Hinman. 1974. Growth, development and composition of the ovine conceptus and mammary gland during pregnancy. J. Anim. Sci. 38:613. Robertson, H. A., J.S.D. Chan and H. G. Friesen. 1980. The use of a pregnancy-specific antigen, chorionic somatomammotrophin, as an indicator of pregnancy in sheep. J. Reprod. Fertil. 58:279. Terqui, M. and C. Delouis. 1975. Les oestrogenes au cours de la gestation et de la parturition. Ieres journees de la Recherche ovine et caprine, INRA, ITOVIC, Paris. Thimonier, J., M. Bosc, J. Djiane, J. Martal and M. Terqui. 1977. Hormonal diagnosis of pregnancy and number of fetuses in sheep and goats. In Management of Reproduction in Sheep and Goats Symposium. Univ. o f Wisconsin, Madison.

Citations

This article has been cited by 2 HighWire-hosted articles: http://www.journalofanimalscience.org/content /53/4/1077#otherarticles