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 was estimated as 0–377 (SE = 0.031), which is comparable with human RFLP heterozygosities for loci chosen in the same way that we selected sheep loci.     

Erythrocyte oxidized glutathione in Australian Merino sheep

The concentration of GSSG was determined in the erythrocytes of Merino sheep. These sheep were grouped according to erythrocyte potassium type, haemoglobin type, and GSH type. It was found that haemoglobin and potassium type were not correlated with GSSG concentration; however, GSSG concentration was found to be significantly correlated with GSH concentration. This relationship may explain previously reported differences in ATPase activity and may reflect further metabolic differences in the erythrocytes of GSH-high and GSH-low type Merino sheep.     

Erythrocyte oxidized glutathione in Australian Merino sheep.

The concentration of GSSG was determined in the erythrocytes of Merino sheep. These sheep were grouped according to erythrocyte potassium type, haemoglobin type, and GSH type. It was found that haemoglobin and potassium type were not correlated with GSSG concentration; however, GSSG concentration was found to be significantly correlated with GSH concentration. This relationship may explain previously reported differences in ATPase activity and may reflect further metabolic differences in the erythrocytes of GSH-high and GSH-low type Merino sheep.     

Volatiles from Merino fleece evoke antennal and behavioural responses in the Australian sheep blow fly Lucilia cuprina

To identify flystrike‐related volatile compounds in wool from Merino sheep, the attractiveness of wool to Lucilia cuprina Wiedmann (Diptera: Calliphoridae) was examined. First, a selection of wool samples guided by previous knowledge of sheep lines, predicted to be more susceptible or more resistant to flystrike, was tested. The attractiveness of the 10 samples selected was not associated with field susceptibility: two samples from the more resistant line were identified as most attractive and two samples from the more susceptible line were identified as least attractive, based on the behavioural assays with gravid flies. Comparison of the headspace volatiles of these samples, using solid phase microextraction and gas chromatography‐mass spectrometry‐electroantennographic detection, revealed octanal and nonanal to be present in the attractive wool samples that elicited responses from the fly antenna. Furthermore, the two compounds were not present in wool that was least attractive to L. cuprina. In laboratory bioassays, octanal and nonanal evoked antennal and behavioural responses in gravid L. cuprina, thus confirming their potential role as semiochemicals responsible for attracting L. cuprina to Merino sheep.     

Carbonic anhydrase activity in blood of early sheep fetuses

• 1.1. The level of carbonic anhydrase activity in the red cells was measured in sheep fetuses at different times after conception: 39, 56, 77, 90 and 140 days, the last being close to full term. Measurements were also made on blood from four of the mothers.
• 2.2. There was a low level of the enzyme present in the 39 day fetuses (0.037 enzyme units (E.U.)/100 μg Hb) and its increase up to 90 days of gestation (0.19 E.U./100 μg Hb) had a form approximating exponential.
• 3.3. The earliest levels were only 11% of the full term levels and only 4% of the adult levels previously reported.
• 4.4. Even the earliest samples were of blood that was fetal rather than embryonic but these results are the earliest carbonic anhydrase activities reported in this mammal.

     

Occurrence of haploid and haploid/diploid mosaic embryos in untreated and androstenedione-immune Australian Merino sheep

Chromosome counts were obtained from 73 out of 177 (41%) early cleavage-stage Merino embryos. A further 13 embryos were classified as probably diploid. Chromosome aberrations were found in 8 (11%) embryos, one of which was aneuploid and the remainder (9.6%) had euploid abnormalities. If the probable diploid embryos are included, the incidence of euploid aberrations falls to 8.1%. Of the abnormal embryos there was one aneuploid with 2N = 55, two haploids, four haploid/diploid mosaics and one zygote with 4 haploid metaphase plates. Two additional zygotes had 4 interphase pronuclei. Four of the euploid abnormalities were attributable to the entry of two or more spermatozoa and therefore polyspermy is the largest single factor leading to chromosomally aberrant embryos in this population of Merino ewes.     

Structure of the epidermis of Australian Merino sheep over a 12-month period

Light-microscopic examination of frozen sections of skin taken from the dorsal thoraco-lumbar region of Australian Merino sheep in winter revealed that the thickness of the epidermis plus a sudanophilic layer was 24.9 micron in the interfollicular region. The uncornified epidermis (10.9 micron) was separated from the sudanophilic layer (14.0 micron) by a thin stratum corneum. It was concluded that the bulk of the sudanophilic layer was emulsified sebum in which was embedded a disorganized collection of desquamated cornified cells. Although large variances were observed in the thickness of the uncornified epidermis and of the sudanophilic layers between sheep and both within the between blocks of tissue obtained from individual sheep, there were no strong seasonal effects on either epidermal structure or layer thickness over a 12-month period. These results suggest that the Australian Merino differs from Finnish Landrace X Dorset Horn ewes, which are reported to possess, at least in winter, a thicker uncornified epidermis and a thicker stratum corneum that could be divided into two zones and was uniformly permeated by lipid.     

Genetic polymorphism in Spanish Merino sheep

A Spanish Merino sheep population is characterized, for the first time, according to its frequencies for a total of nine polymorphic loci: three blood group factor systems, A, B and C, and the following red cell or serum polymorphisms: haemoglobin (Hb), carbonic anhydrase (CA), 'X protein', transferrin (Tf), arylesterase (EsA) and albumin (Al). Another locus, amylase (Am), did not show polymorphism.     

A large-scale program in laparoscopic intrauterine insemination with frozen-thawed semen in Australian Merino sheep in Argentine Patagonia

This report shows the results of a large-scale laparoscopic intrauterine insemination program on a flock of Australian Merino sheep in Argentine Patagonia. The study was carried out on a total of 1824 ewes (3-to-7-yr-old) and 480 ewe hoggets (19-20 months old) on 2 farms in the southeastern region of Santa Cruz Province, in April and May 1996. The animals, divided into 15 groups, were synchronized with vaginal sponges containing 60 mg medroxyprogesterone acetate for 14 d and injected with 200 IU PMSG upon sponge removal. Estrus was screened every 12 h by means of vasectomized marker rams. The animals were inseminated laparoscopically by the intrauterine route using 2 schemes: 1) at a fixed time (12 h) after estrus detection, or 2) at a fixed time (60 h) after sponge removal irrespective of estrus. Pregnancy was determined at 30 d by transrectal ultrasound imaging. The results showed that 1) the onset of estrus occurs most often between 24 and 48 h after sponge removal, 2) ewe hoggets undergo estrus significantly earlier than sexually mature ewes, 3) in those animals showing estrus, there appears to be no relationship between fertility (as assessed by pregnancy outcome) and time of estrus, 4) there is a significant association between the percentage of estrus occurrence and pregnancy rate, 5) fertility is significantly higher in ewes than in hoggets, 6) for practical purposes insemination at a fixed time after the onset of estrus has no advantage over that of to insemination at a fixed time after sponge removal. It is concluded that large-scale laparoscopic intrauterine insemination can be successfully applied in Australian Merino ewes and ewe hoggets in low-productivity areas such as that of Argentine Patagonia and that estrus detection is unnecessary when insemination is performed at 60 h after sponge removal.     

Uterine presensitization and embryo survival and growth in Booroola Merino x South Australian Merino ewes

The fertility enhancing effects of semen were examined following the intra-uterine insemination of killed spermatozoa plus seminal plasma 17 d prior to insemination with viable spermatozoa. Three experiments were conducted: two on 1.5-yr old and 2.5 to 5.5 yr-old Booroola Merino x South Australian Merino ewes in 1986 and one on 1.5 yr-old ewes in 1987. Differences between treatment and control groups for the percentage of ewes exhibiting estrus by Days 21 and 35 following fertile insemination, the percentage of ewes with viable embryos at Day 35, the number and weight of viable embryos per ewe, the nubmer of caruncular implantation sites and the progesterone level were not statistically significant (P>0.05). There were no statistically significant treatment by experiment interactions for any of the variables examined. Inflammation and edema of the endometrial tissue was not observed following the presensitization treatment.     

A single nucleotide polymorphism on chromosome 10 is highly predictive for the polled phenotype in Australian Merino sheep

The aim of this study was to fine map the genomic location of the Horns locus in the Australian Merino sheep population and to identify markers that can be used to predict the horn phenotype. A linkage disequilibrium analysis of horn data from Australian Merino sheep mapped the Horns locus to a small region on chromosome 10. A single nucleotide polymorphism in the region was found to be highly predictive for the polled phenotype in an experimental population of Merino sheep. This was owing to a dominance effect of one of the alleles when inherited maternally. It was suggested that a genetic test would provide a good predictor of the polled phenotype. Finally, an evaluation of industry data showed that the SNP is at very different frequencies in Poll Merino sheep that have been bred for polledness (based on phenotype alone) compared with the Merino sheep breed.     

Piglets born from centrifuged and vitrified early and peri-hatching blastocysts

Cryopreservation of zona-intact porcine embryos has been relatively unsuccessful to date, although some success has been obtained with lipid reduced morulae and early blastocysts. This study adapted some vitrification protocols used successfully with late blastocysts for use with early zona-intact blastocysts, using actin depolymerization, centrifugation, and open-pulled (OPS) straws. Initially, Day 6 peri-hatching blastocysts were collected, cultured for 40 min in 7.5 microg/ml cytochalasin B and vitrified in 6.5 M glycerol and 6% BSA (VS1) in either heat-sealed (HS) or open straws (OS). The post-thaw survival of those stored in HS was 15.4% after 24 and 48 h in vitro; storage in OS significantly improved survival (58.8% for both 24 and 48 h). When similar stage blastocysts were cultured in cytochalasin B and vitrified with 8 M ethylene glycol and 7% polyvinylpyrrolidone (PVP; VS2) in OS, survival was 44.4 and 33.3% for 24 and 48 h, respectively. Day 5 late morulae and early blastocysts were collected, cultured with cytochalasin B, and centrifuged or left intact (control), then vitrified with VS1 in HS or OS, or vitrified in VS2 in OS only. None of the intact control embryos survived thawing and 48 h culture in vitro. Centrifuged early blastocysts vitrified with VS1 showed good post-thaw survival in culture when stored in HS (62.8 and 60.5% for 24 and 48 h, respectively), or OS (75 and 63.6%). When vitrified with VS2 in OS, survival improved (80 and 76.7%). Peri-hatching blastocysts were vitrified in VS1, and early blastocysts were vitrified with VS1 and VS2. All blastocysts were stored in OS. The embryos were recovered and transferred to Day 4 and 5 pseudopregnant recipients (for Day 5 and 6 blastocysts, respectively). Of the five recipients receiving peri-hatching blastocysts, two became pregnant and delivered a total of eight piglets. All three recipients of early blastocysts vitrified in VS1 had a delayed return to estrus; while of the four receiving embryos vitrified with VS2, two were delayed in returning to estrus, and one was confirmed pregnant after 45 days. A litter of five piglets, one male and four female, was produced at 116 days of gestation. To our knowledge, this is the first litter of piglets produced from early blastocysts vitrified without micromanipulation to remove polarized lipid droplets.     

Factors influencing lamb survival in a high fecundity Booroola Merino x South Australian Merino flock

The effects of year of lambing, age of ewe and litter size on lamb survival and birthweight, and the effects of ewe mating weight and pregnancy wastage (ovulation rate minus litter size) on birthweight were examined in Booroola Merino x South Australian Merino ewes. Year of lambing, litter size and their interaction were significant (P<0.001) sources of variation for lamb survival. When birthweight was included as a linear and quadratic covariate for lamb survival, year of lambing and litter size and their interaction remained as statistically significant sources of variation. Year of lambing, litter size and pregnancy wastage contributed significantly (P<0.05) to variation in birthweight. Ewe liveweight at mating was not an important source of variation (P>0.05). Birthweight was significantly (P<0.05) reduced with an increase in pregnancy wastage. Improvement of birthweight of multiple birth lambs has some potential for increasing lamb survival. Other factors influencing lamb survival (year of lambing, litter size, pregnancy wastage) require further study so that strategies for reducing lamb loss in high fecundity Merino flocks can be developed.     

Polyploid abnormalities in day 3 and day 5 merino sheep embryos

The chromosome complement was assessed in Merino sheep embryos collected at 3 and 5 days after the onset of oestrus. Donor ewe treatments were: untreated, or immunized against androstenedione (day 3); and untreated, or treated with follicle-stimulating hormone (FSH), or treated with FSH plus immunization against androstenedione (day 5). No significant differences in the frequency of chromosomally abnormal embryos between treatment groups within each age group were observed, so the data have been combined. Euploid abnormalities were observed in 10.8% of the day-3 embryos (4/37), with the abnormalities being one 1n, one 3n and two 5n. Embryos with euploidy (10%) were also observed at day 5, with three 1n/2n mosaics and a 3n embryo present in a sample of 40. These data suggest that chromosomally aberrant embryos are not lost before day 5 of development.     

Fertilization and embryo loss in Booroola Merino x South Australian Merino ewes: Effect of the F gene

The effects of Booroola genotype (F+, ++); the number of ovulations per ewe (one, two or three); and the age of a ewe (2.5 yr vs 3.5 to 6.5 yr) on the percentage of ova fertilized, embryo loss and fetal loss were examined in Booroola x South Australian Merino ewes slaughtered on Days 4, 21 and 90 after insemination. Ewes slaughtered on Day 90 were examined by real-time ultrasound imaging (RUI) on Day 45. Fertilization failure was independent of ewe genotype, ovulation rate and age of ewe, and it was not an important source of wastage (F+, 9.4%; ++, 6.7%). Most embryo loss occurred during the first 21 d (F+, 54.7%; ++, 40.3%). Interpretation of the effects of genotype and ovulation rate on embryo wastage measured on Days 21, 45 and 90 was obscured by significant (P < 0.05) genotype and ovulation rate interactions with the day of slaughter/RUI. The effect of age on embryo loss was not significant (P > 0.05). Reasons for the high rate of wastage observed in this experiment require further study.