Direct Evidence on the Contribution of a Missense Mutation in GDF9 to Variation in Ovulation Rate of Finnsheep

peer-reviewedThe Finnish Landrace (Finnsheep) is a well known high-prolificacy sheep breed and has been used in many countries as a source of genetic material to increase fecundity of local breeds. Analyses to date have indicated that mutations with a large effect on ovulation rate are not responsible for the exceptional prolificacy of Finnsheep. The objectives of this study were to ascertain if: 1) any of 12 known mutations with large effects on ovulation rate in sheep, or 2) any other DNA sequence variants within the candidate genes GDF9 and BMP15 are implicated in the high prolificacy of the Finnish Landrace breed; using material from lines developed by divergent selection on ovulation rate. Genotyping results showed that none of 12 known mutations (FecBB, FecXB, FecXG, FecXGR, FecXH, FecXI, FecXL, FecXO, FecXR, FecGE, FecGH, or FecGT) were present in a sample of 108 Finnsheep and, thus, do not contribute to the exceptional prolificacy of the breed. However, DNA sequence analysis of GDF9 identified a previously known mutation, V371M, whose frequency differed significantly (P<0.001) between High and Low ovulation rate lines. While analysis of ovulation rate data for Finnsheep failed to establish a significant association between this trait and V371M, analysis of data on Belclare sheep revealed a significant association between V371M and ovulation rate (P<0.01). Ewes that were heterozygous for V371M exhibited increased ovulation rate (+0.17, s.e. 0.080; P<0.05) compared to wild type and the effect was non-additive (ovulation rate of heterozygotes was significantly lower (P<0.01) than the mean of the homozygotes). This finding brings to 13 the number of mutations that have large effects on ovulation rate in sheep and to 5, including FecBB, FecGE, FecXO and FecXGR, the number of mutations within the TGFβ superfamily with a positive effect on prolificacy in the homozygous state

Effects of extended grazing during mid, late or throughout pregnancy, and winter shearing of housed ewes, on ewe and lamb performance

peer-reviewedA flock of March-lambing ewes was used to evaluate the effects of (i) extended (deferred, winter) grazing of pasture during mid, late or throughout pregnancy, and (ii) winter shearing of ewes housed during mid and late pregnancy, on lamb birth weight and subsequent growth to weaning. Ewes (n = 265) were allocated at random to five treatments for the period from 7 December (~ day 47 of pregnancy) to lambing. The treatments were: housed shorn (HS), housed unshorn (HU), grazing throughout (EG), grazing to 20 January followed by housing (EGH), housed to 20 January followed by grazing (HEG). From 1 March to lambing the HEG and EG ewes were dispersed on the paddocks intended for grazing post lambing. All ewes were offered a concentrate supplement during the final 6 weeks of pregnancy. Housed ewes were offered grass silage while ewes on extended grazing were allocated 1.3 kg herbage dry matter per head per day from swards that had been closed for approximately 10 weeks. Ewes plus lambs (except triplet-rearing ewes which were grazed separately) from all treatments were grazed together post lambing, grouped according to lambing date. For treatments HS, HU, EGH, HEG and EG gestation lengths were 147.0, 145.6, 146.3, 146.6 and 146.9 (s.e. 0.34, P < 0.001) days, lamb birth weights were 4.9, 4.3, 4.4, 4.6 and 5.0 (s.e. 0.10, P < 0.001) kg, and lamb weaning weights were 34.6, 32.1, 33.3, 33.8 and 34.9 (s.e. 0.66, P < 0.001) kg, respectively. Extended grazing in mid and late pregnancy resulted in 35% and 65%, respectively, of the increase in lamb birth weight associated with extended grazing throughout. Treatment effects on lamb birth weight were associated with those on weaning weight (P < 0.01, R2 = 0.93). It is concluded that extended grazing or shearing of housed ewes increased lamb birth weight and subsequent weaning weight. The increased lamb birth weight from deferred grazing in mid pregnancy was probably due to improved protein utilisation from the grazed herbage. Meanwhile, the increase

Genetic and non-genetic factors affecting lamb growth and carcass quality.

End of Project ReportsThe work undertaken under this project concerned the effects of genetic and non-genetic factors on lamb growth, both pre and post-weaning, and carcass traits. The principal objective of the genetic studies was to estimate the performance effects of selecting terminal sires on the basis of the lean meat index (LMI) which is produced for pedigree lambs in flocks that participate in the national Breed Improvement Programme operated by the Department of Agriculture and Food. The merits of the Beltex breed, recently introduced to this country, were also evaluated on comparisons with Texel and Suffolk sires. Estimates of within-breed genetic variation for growth and carcass traits were obtained.European Union Structural Funds (EAGGF

An evaluation of two grassland-based systems of mid-season prime lamb production using prolific ewes of two genotypes

peer-reviewedA 4-year study was undertaken to evaluate the effects of two contrasting management systems [year-round grazing (YRG) and normal seasonal grazing followed by indoor feeding during winter (GWF)] on performance of mid-season lambing ewes. On the GWF system, the annual stocking rate was 14.4 ewes/ha, grass silage was conserved for winter feeding indoors, and the ewes were lambed indoors and were then turned out to pasture. The YRG system was stocked at 10.5 ewes/ha, was grazed during the winter, had outdoor lambing and the animals had access to all the farmlet for summer grazing. The ewes were Belclare and Cheviot × Belclare which were balanced across systems. Mean lambing dates and fertiliser N application rates were 20 and 30 March, and 85 and 92 kg/ha, for the GWF and YRG systems, respectively. Concentrate supplementation during late pregnancy was similar on both systems. For the GWF and YRG systems, litter size, lamb mortality, number of lambs reared, birth weight (kg), weaning weight (kg) and lamb carcass output (kg/ha) were 2.17 and 2.24 (s.e. 0.038), 10.1 and 13.8% (P = 0.05), 1.77 and 1.78 (s.e. 0.042), 4.0 and 4.7 (s.e. 0.05, P < 0.001), 27.9 and 30.8 (s.e. 0.25, P < 0.001) and 469 and 348, respectively. Belclare ewes had a higher litter size (2.34 v 2.07; s.e. 0.038, P < 0.001) and number of lambs reared per ewe joined (1.86 v 1.69; s.e. 0.048, P < 0.01) than the Cheviot × Belclare ewes. There were no significant interactions between system and ewe breed type. It is concluded that the YRG system of prime lamb production was sustainable using prolific ewes but at a reduced stocking rate (−26%) and with greater lamb mortality relative to the GWF system. Ewe genotypes with a mean litter size of up to 2.34 lambs are suitable for both systems. Lamb carcass output of 501 kg/ha was achieved from a primarily grass-based system of mid-season prime lamb production using prolific ewes (Belclare)

A note on the use of FTA™ technology for storage of blood samples for DNA analysis and removal of PCR inhibitors

peer-reviewedFTA™ technology is widely used across many molecular disciplines for sample capture, storage and analysis. The use of this technology for the long-term storage of blood samples for DNA analysis was examined as well as its potential to remove inhibitors from DNA samples previously extracted from blood with PCR inhibitors remaining. It was found that blood spots stored on FTA™ cards for 8 years at room temperature gave successful PCR products and that FTA™ cards are a useful tool for removing substances in samples which interfere with or inhibit, the PCR reaction

Investigation of Prolific Sheep from UK and Ireland for Evidence on Origin of the Mutations in BMP15 (FecXG, FecXB) and GDF9 (FecGH) in Belclare and Cambridge Sheep

peer-reviewedThis paper concerns the likely origin of three mutations with large effects on ovulation rate identified in the Belclare and Cambridge sheep breeds; two in the BMP15 gene (FecXG and FecXB) and the third (FecGH) in GDF9. All three mutations segregate in Belclare sheep while one, FecXB, has not been found in the Cambridge. Both Belclare and Cambridge breeds are relatively recently developed composites that have common ancestry through the use of genetic material from the Finnish Landrace and Lleyn breeds. The development of both composites also involved major contributions from exceptionally prolific ewes screened from flocks in Ireland (Belclare) and Britain (Cambridge) during the 1960s. The objective of the current study was to establish the likely origin of the mutations (FecXG, FecXB and FecGH) through analysis of DNA from Finnish Landrace and Lleyn sheep, and Galway and Texel breeds which contributed to the development of the Belclare breed. Ewes with exceptionally high prolificacy (hyper-prolific ewes) in current flocks on Irish farms were identified to simulate the screening of ewes from Irish flocks in the 1960s. DNA was obtained from: prolific ewes in extant flocks of Lleyn sheep (n = 44) on the Lleyn peninsula in Wales; hyper-prolific ewes (n = 41); prolific Galway (n = 41) ewes; Finnish Landrace (n = 124) and Texel (n = 19) ewes. The FecXG mutation was identified in Lleyn but not in Finnish Landrace, Galway or Texel sheep; FecXB was only found among the hyper-prolific ewes. The FecGH mutation was identified in the sample of Lleyn sheep. It was concluded from these findings that the Lleyn breed was the most likely source of the FecXG and FecGH mutations in Belclare and Cambridge sheep and that the FecXB mutation came from the High Fertility line that was developed using prolific ewes selected from commercial flocks in Ireland in the 1960′s and subsequently used in the genesis of the Belclare.Financial support through the Teagasc Walsh Fellowship Scheme, Genesis Faraday SPARK award (Lleyn survey) and Science Foundation Ireland (07/SRC/B1156) is gratefully acknowledged

Response to Teladorsagia circumcincta infection in Scottish Blackface lambs with divergent phenotypes for nematode resistance

peer-reviewedThe objective of this study was to identify Scottish Blackface lambs that were at the extremes of the spectrum of resistance to gastrointestinal nematodes and characterise their response to an experimental nematode challenge. Lambs (n = 90) were monitored for faecal egg count (FEC) (2 samples from each of 2 independent natural infections). The most resistant (n = 10) and susceptible (n = 10) individuals were selected and challenged with 30,000 Teladorsagia circumcincta larvae (L3) at 9 months of age. Response to infection was monitored by measuring FEC, plasma pepsinogen, serum antibodies against nematode larval antigens and haematology profile, until necropsy at 71 days post infection. Worm burden, worm fecundity and the level of anti-nematode antibodies in abomasal mucosa were determined at necropsy. FEC was consistently higher in susceptible animals (P < 0.05), validating the selection method. Worm fecundity was significantly reduced in resistant animals (P = 0.03). There was also a significant correlation (r = 0.88; P < 0.001) between the number of adult worms and FEC at slaughter. There was no effect of phenotype (resistance/susceptibility) on plasma pepsinogen or on haematology profile. Phenotype had a significant effect on the level of anti-nematode IgA antibodies in serum (P < 0.01), reflecting a higher peak in resistant animals at day 7 post infection. It is concluded that significant variation in the response to gastrointestinal nematode challenge exists within the Scottish Blackface population with resistant animals displaying significantly lower FEC, lower worm fecundity and higher concentration of anti-nematode IgA antibodies in serum.Kathryn McRae was supported by a Teagasc Walsh fellowship and the Allan and Grace Kay Overseas Scholarship

Gastrointestinal nematode control practices on lowland sheep farms in Ireland with reference to selection for anthelmintic resistance

Gastrointestinal parasitism is a widely recognised problem in sheep production, particularly for lambs. While anthelmintics have a pivotal role in controlling the effects of parasites, there is a paucity of data on how farmers use anthelmintics. A representative sample of Irish lowland farmers were surveyed regarding their parasite control practices and risk factors that may contribute to the development of anthelmintic resistance. Questionnaires were distributed to 166 lowland Irish sheep producers. The vast majority of respondents treated their sheep with anthelmintics. Lambs were the cohort treated most frequently, the majority of farmers followed a set programme as opposed to treating at sign of disease. A substantial proportion (61%) administered four or more treatments to lambs in a 'normal' year. Departures from best practice in anthelmintic administration that would encourage the development of anthelmintic resistance were observed. In conclusion, in the light of anthelmintic resistance, there is a need for a greater awareness of the principles that underpin the sustainable use of anthelmintics and practices that preserve anthelmintic efficacy should be given a very high priority in the design of helminth control programmes on each farm. To this end, given that veterinary practitioners and agricultural advisors were considered to be the farmer's most popular information resource, the capacity of these professions to communicate information relating to best practice in parasite control should be targeted

AI For Sheep Using Frozen-thawed Semen.

End of Project ReportInternational experience has been that cervical insemination of sheep with frozen-thawed semen usually yields unacceptably low pregnancy rates (10 to 30%). An exceptional case has been Norway where non-return rates in on-farm usage are around 60%. The objective of the work described in this report was to develop an AI procedure for Irish conditions, based initially on Norwegian protocols, using semen from individual rams. Such a procedure would greatly facilitate and enhance genetic improvement programmes for sheep. The work undertaken had two separate aspects:- (i) studies on semen, including processing and freezing methods, laboratory evaluation of semen quality post thawing and the relationship of in vitro evaluation to fertilisation rate in vivo (ii) studies on pregnancy rate following AI in relation to issues such as ram breed effects, effects of synchronisation, operator differences and the role of ewe breed inducing the timing of ovulation and various physical and physiological assessments of the cervix at AI. The main results in relation to semen studies were that, while a range of differential staining procedures could be used to objectively evaluate semen with respect to proportion of live speramatozoa and the integrity of sperm cells after thawing, these results were not useful as indicators of fertilisation capacity in vivo. The in vitro fertilisation (IVF) of sheep oocytes recovered from abattoir material gave promising results as a method for evaluating the fertilisation capacity of frozen-thawed semen. The technique requires further validation.Department of Agriculture, Food and the Marin

The Solvent–Solid Interface of Acid Catalysts Studied by High Resolution MAS NMR

High-resolution magic angle spinning (HRMAS) NMR spectroscopy was used to study the effect of mixed solvent systems on the acidity at the solid−liquid interface of solid acid catalysts. A method was developed that can exploit benefits of both solution and solid-state NMR (SSNMR) by wetting porous solids with small volumes of liquids (μL/mg) to create an interfacial liquid that exhibits unique motional dynamics intermediate to an isotropic liquid and a rigid solid. Results from these experiments provide information about the influence of the solvent mixtures on the acidic properties at a solid−liquid interface. Importantly, use of MAS led to spectra with full resolution between water in an acidic environment and that of bulk water. Using mixed solvent systems, the chemical shift of water was used to compare the relative acidity as a function of the hydration level of the DMSO-d6 solvent. Nonlinear increasing acidity was observed as the DMSO-d6 became more anhydrous. 1H HR-MAS NMR experiments on a variety of supported sulfonic acid functionalized materials, suggest that the acid strength and number of acid sites correlates to the degree of broadening of the peaks in the 1H NMR spectra. When the amount of liquid added to the solid is increased (corresponding to a thicker liquid layer), fully resolved water phases were observed. This suggests that the acidic proton was localized predominantly within a 2 nm distance from the solid. EXSY 1H−1H 2D experiments of the thin layers were used to determine the rate of proton exchange for different catalytic materials. These results demonstrated the utility of using (SSNMR) on solid−liquid mixtures to selectively probe catalyst surfaces under realistic reaction conditions for condensed phase systems