Genetic variation in Australian Merino sheep.

Variation at 22 gene loci was investigated in a flock of Australian Merino sheep using restriction fragment length polymorphism (RFLP) analysis. Polymorphism was observed at 20 loci, including loci for wool keratin, hormone and immunoglobulin light chain genes. Eleven loci yielded unambiguous genotypes suitable for population data analysis. Average heterozygosity, determined from these and two monomorphic loci, was estimated as 0.107 (SE = 0.024). Average heterozygosity excluding all monomorphic data as estimated at 0.377 (SE = 0.031), which is comparable with human RFLP heterozygosities for loci chosen in the same way that we selected sheep loci.

Evidence of linkage between high-glycine-tyrosine keratin gene loci and wool fibre diameter in a Merino half-sib family.

Candidate genes for quantitative trait loci have been studied in a Medium Peppin Merino flock. Obvious candidates for effects on wool production traits are genes for the major proteins expressed in the wool fibre, the keratin and keratin-associated protein genes. Two keratin-associated protein loci, KRTAP6 and KRTAP8, have previously been shown to be linked. The results of analyses between these two loci and production traits gave significant evidence of linkage with wool fibre diameter in one out of eight half-sib groups tested. High-glycine-tyrosine proteins (KRTAP6, 7 and 8) are known to vary considerably in abundance in wool fibres and it is possible that a gene for major effect on fibre diameter is located within the same chromosomal region as KRTAP6 and KRTAP8.

Linkage relationships between keratin-associated protein (KRTAP) genes and growth hormone in sheep.

The ovine hair keratin-associated protein (KRTAP) genes are grouped into several families. In this study, we analyzed linkage relationships between members of four KRTAP families and growth hormone and beta-hemoglobin. Restriction fragment length polymorphisms and a microsatellite polymorphism were typed in 10 sire groups from a flock of Australian Merino sheep. Two linkages, both previously undetected in sheep or any other mammal, were found. They were between growth hormone (GH) and the high-sulfur keratin gene family KRTAP1 (previously called high-sulfur B2C) at theta = 0.10, Z = 3.01 and between two high-glycine-tyrosine class genes, KRTAP6 (previously high-glycine-tyrosine type II) and KRTAP8 (previously high-glycine-tyrosine type I-F), at theta = 0.15, Z = 5.0. The high level of conservation of the mammalian genome at these recombination values suggests that these linkages might be found in distantly related mammalian species.

Genetic markers for the Booroola fecundity (Fec) gene in sheep.

Animals from the Booroola line of Australian Merino sheep are characterized by a high ovulation rate that can be attributed to the presence of a codominant allele (FecB). The specific function of the gene has not been identified. Effective use of the trait within the sheep breeding industry requires one or more genetic markers that can distinguish between alternative alleles at the locus Fec. With a combination of DNA minisatellite markers and polymorphic protein markers, a cluster of seven minisatellite fragments has been identified as being linked to the Fec gene and to the ovine A blood group locus. The minisatellite fragments have been derived from multilocus probes and hence cannot be used to define the chromosomal location of the Fec gene or to serve as diagnostic markers for Fec. The derivation of cloned single locus markers from the minisatellite fragments will enable finer scale mapping of the Fec and the A blood group locus in sheep.

Response to helminth infection of sheep selected for resistance to Haemonchus contortus.

Lines of Merino sheep selected for increased (IRH) and decreased (DRH) resistance to Haemonchus contortus were compared with an unselected (CH) line, after approximately four generations of selection. Measurements were recorded on 69 IRH, 47 DRH and 84 CH animals. Following artificial challenge with H. contortus, the IRH line had significantly (P less than 0.001) lower faecal egg counts than the CH and DRH lines (2730, 12,720 and 17,400 epg, respectively). Significant differences (P less than 0.05) were found between all lines in the minimum packed cell volumes during artificial infection (25.7, 22.0 and 20.3%) and in faecal egg counts after natural infection (140, 3590 and 8750 epg). Differences were also recorded (P less than 0.05) following artificial challenge with Trichostrongylus colubriformis (490, 840 and 1340 epg). On a percentage basis, faecal egg counts in the IRH line deviated less from the CH line following artificial infection with T. colubriformis (42%) than with H. contortus (79%). The reverse was true for the DRH line (60 and 37%, respectively). Differences in egg output of this magnitude should have a marked effect on requirements for anthelmintic treatment, rate of development of drug resistance and level of pasture contamination when the lines are grazed separately.

Ovarian follicular dynamics in lines of sheep (Finn, Merinos) selected on ovulation rate.

Ewes from selected lines of sheep from each of two breeds, Finns (high ovulation rate, low ovulation rate and control lines with respective ovulation rates of 5.4, 2.7 and 3.3) and Merinos (T Merinos selected for increased ovulation rate and control Merinos with respective ovulation rates of 1.9 and 1.2) were used to examine how selection to alter ovulation rate had altered follicle development. Ovarian antral follicles were counted, measured, classified as nonatretic or atretic (more than five pyknotic bodies). The growth of ovulatory follicles in vivo, followed by repeated follicle ink marking, also was compared in the three lines of Finns. Regardless of breed, ewes selected for high ovulation rate had a similar number of antral follicles and a similar extent of atresia compared with their controls. Alterations induced by selection were located in the last stages of folliculogenesis. T Merinos exhibited a lower proportion of atretic follicles among follicles greater than 3 mm and a larger diameter of the largest healthy follicle when preovulatory follicles were excluded. High-line Finn ewes recruited more follicles, which produced smaller preovulatory follicles, each containing a smaller number of granulosa cells compared with either the low- or control-line ewes. Hence, physiological selection for high ovulation rate raised it by different methods in Merino than in Finn ewes.

No association between the ovine leucocyte antigen (OLA) system in the Australian merino and susceptibility to Haemonchus contortus infection.

A genetic analysis has been made of the Ovine Leucocyte Antigenic (OLA) system in Australian merinos. The animals consisted of sires, dams and their progeny. The typing data were consistent with previous findings of a high degree of polymorphism. At least two closely linked loci with several alleles at each are necessary to explain the data. No evidence was found for an association between OLA types and three measures of susceptibility to infection by the blood-sucking parasite Haemonchus contortus. Attention is drawn to the utility of half-sib data for analysis of the genetic control of resistance to parasites in sheep and other animals with a similar breeding structure.

Increase in ovulation rate in Merino ewes after active immunization with inhibin preparations obtained by immunoaffinity chromatography.

Ewes were immunized with four inhibin preparations of increasing purity obtained from bovine follicular fluid by affinity chromatography. The two purest immunogens, which had 69 and 174 ng inhibin micrograms-1 protein (expressed in terms of 32 kDa ovine inhibin), increased ovulation rate by 240 and 320% respectively following two injections, and gave a transient increase in plasma FSH concentration at the same time. Plasma antibodies recognizing native 125I-31 kDa bovine inhibin appeared at the same time, and, together with the increased ovulation rate, persisted after the injections ceased. Ovulation rate and antibody titres were raised by the less pure immunogens, but to a smaller extent, and the increases did not persist. It was concluded that the increased ovulation rate was due to induced inhibin antibodies, acting at least in part through the consequent rise in plasma FSH concentration.

Induction of ovulation in chronically hypophysectomized Booroola ewes.

Booroola Merino ewes, with (F+; N = 17) and without (++; N = 13) a copy of the fecundity gene were hypophysectomized and 6 weeks later were given an i.m. injection of PMSG (high, medium or low dose) followed by hCG. The induced ovulation rates were observed laparoscopically. Ovulation rates were significantly higher (P less than 0.01) in Booroola F+ ewes than in ++ ewes (8.00 +/- 1.66 s.e.m. vs 3.62 +/- 1.10 respectively). This suggests that the high fecundity of the Booroola ewe may be due primarily to ovarian rather than pituitary factors.

The half-life of follicle-stimulating hormone in ovary-intact and ovariectomized booroola and control merino ewes.

To determine whether the high ovulation rate of the Booroola Merino ewe could be explained by FSH metabolism we have tested the proposition that FSH may have a longer half-life in the plasma of Booroola Merino ewes than in control ewes. The half-life of plasma FSH was determined by removal of the pituitary gland, to abolish FSH secretion into the peripheral circulation, and monitoring by repeated blood sampling the subsequent decline in plasma FSH concentrations. The half-life of FSH was similar in Booroola (103 +/- 14 (s.e.m.) min, N = 8) and control (116 +/- 8 min, N = 9) ewes. However, when ewes that had been ovariectomized at least 6 months earlier were hypophysectomized, the half-life of FSH was increased from 110 + 8 min in ovary-intact ewes (N = 11) to 1101 +/- 49 min (N = 6) (P less than 0.001) with no difference between the two Merino strains. We conclude that changes in the circulating half-life of FSH do not account for the high fecundity of the Booroola but that ovariectomy can alter the half-life of FSH secreted by the pituitary gland.

Suppression of ovine plasma FSH by bovine follicular fluid: neutralization by plasma from ewes immunized against an inhibin-enriched preparation from bovine follicular fluid.

Adult Merino ewes were immunized against an inhibin-enriched preparation (bFFI) obtained by affinity chromatography of bovine follicular fluid (bFF). Plasma was obtained in early luteal phase from these ewes and from control ewes immunized against bovine serum albumin. Ten months after ovariectomy the plasma concentration of FSH, but not LH, in control ewes was decreased by four s.c. injections of 8 ml bFFI (17,500 units inhibin/injection). There was no decrease in plasma concentrations of FSH or LH in immunized ewes with the same dose of bFFI. In a second study with long-term ovariectomized ewes, four injections of 20 ml plasma from the immunized ewes significantly reduced the decrease in FSH concentration caused by four injections of steroid-free bFF (2500 units inhibin/injection) in comparison with similar ewes injected with plasma from control ewes. These results show that the plasma of ewes immunized against bFFI contains substances, presumably antibodies, which neutralize the FSH-suppressive effects of bFF and bFFI in ovariectomized ewes.