甘肃肉羊新品种选育群GDF9基因外显子2多态性及其与产羔性状关联分析

根据GenBank发布的绵羊GDF9基因外显子2的序列设计4对引物,采用PCR-SSCP技术分析GDF9基因外显子2在甘肃内羊新品种选育群羊中的单核苷酸多态性,并与产羔性状进行关联分析.结果表明,GDF9基因的扩增片段在所检测的新品种群羊中存在PCR-SSCP多态性,检测到3种基因型(AA、AB和BB),而在32只无角陶赛特母羊群中只检测到AA和AB基因型.测序结果显示,GDF9基因编码区第978位碱基发生A→G突变,但没有导致氨基酸的改变;第994位碱基发生G→A突变,导致Ⅴ变成Ⅰ(缬氨酸→异亮氨酸).新品种选育群羊产羔数的最小二乘均值关系为AB＞ AA＞ BB,统计分析结果初步表明3种基因型之间差异不显著(P＞0.05).故该区域可能不是影响新品种群羊繁殖力的功能结构区城.     

乐至黑山羊PRLR基因外显子10多态性与产羔数的关系研究

设计2对特异性引物对乐至黑山羊PRLR基因第10外显子进行了PCR-SSCP检测,并研究该基因与产子性能的相关性。结果表明,P1引物扩增片段不存在多态性;P2引物扩增片段存在多态性,表现为AA,AB,AD和CD 4种基因型,测序结果表明,4种基因型都在该片段第89、94、146和157位存在C→T、A→C、C→G、G→C的突变;此外AA型还在61位发生C→T的突变;AD型还在175位发生A→G的突变;CD型还在24位发生T→C的突变,96位发生C→T的突变,通过统计分析发现AD型平均产羔数优于其他3种基因型,并且与AB型差异达到显著水平(P<0.05)。因此认为PRLR基因对于乐至黑山羊产子性能有一定的影响。     

Second litter syndrome in Iberian pig breed: factors influencing the performance

Second litter syndrome (SLS) consists of a loss of prolificacy in the second parity (P2), when a sow presents the same or lower results for litter size than in the first parity (P1). This syndrome has been reported for modern prolific breeds but has not been studied for rustic breeds. The objectives of this study are to determine how and to what degree Iberian sows (a low productivity breed recently raised on intensive farms) are affected by SLS; to establish a target and reference levels; and to assess the factors influencing the performance. Analysed data correspond to 66 Spanish farms with a total of 126 140 Iberian sows. The average Iberian sow prolificacy in P1 was 8.91 total born (TB) and 8.47 born alive (BA) piglets, whereas in P2, it decreased by ?0.05 TB and ?0.01 BA piglets, suggesting some general incidence of SLS. At the sow level, 56.63% did not improve prolificacy in terms of BA piglets in P2, and 16.98% had a clear decrease in prolificacy, losing ≥3 BA piglets in P2. Within herds, a mean of 57.75% of sows showed SLS, with an evident decrease in the number of BA piglets in P2. The plausible target for the Iberian farm’s prolificacy comes from the quartile of farms with the lowest percentage of SLS sows within the farms with the highest prolificacy between P1 and P2 (mean of 8.77 BA). So, in this subset of farms (N = 17), 47.3% of sows improved their prolificacy in P2 (i.e. did not show SLS). Hence, half the sows could be expected to show SLS even on farms with a good performance. Finally, this study brings out the main factors reducing P2 prolificacy through SLS in the Iberian breed: later age at first farrowing, long first lactation length, medium weaning to conception interval and large litter size in P1. In conclusion, improving the reproductive performance of Iberian farms requires reducing the percentage of sows with SLS, paying special attention to those risk factors. The knowledge derived from this study can provide references for comparing and establishing objectives of performance on Iberian sow farms which can be used for other robust breeds.     

Improving productivity of Malpura breed by crossbreeding with prolific Garole sheep in India

Garole, a prolific small sized sheep breed of India was introduced in the Mutton Project of the Institute during 1997 to explore the possibility of incorporating fecundity gene into Malpura (M) sheep of semi-arid region of Rajasthan. The data on reproduction and production traits of M and Garole × Malpura (GM) half-breds was used in the study to assess the improvement in evolved crossbred stock. The overall percent gain in prolificacy in GM was 52.38, which increased to 75.73 in third parity. The GM compared to M ewes also weaned more lambs per ewe lambed (1 versus 1.41). The lambing rate averaged 91.75 and 148.21% in M and GM ewes, respectively. The twin lambing percentage was only 4.71% in M flock while; it reached up to 51.10% in GM. Moreover, 5.72% of GM ewes produced triplets also. The genetic group non-significantly affect ewe efficiency (EE), but GM produced 3.3% more total lambs weight per ewe lambed at weaning and 6.15% more total lamb weight per ewe lambed at 6-month of age. The body weights at different ages of GM half-breds were lower (P < 0.01) compared to contemporary M lambs. The parity of ewes significantly affects body weight from birth weight to 12-month of age. The survivability of GM half-breds was almost at par with the local M sheep. Results revealed that the fecundity genes responsible for increasing prolificacy have been incorporated into the GM and it might prove a valuable germplasm towards evolving a new prolific strain of sheep. The crossing of Garole with M increased the ewe efficiency.     

Prevention of suckling improves postpartum reproductive responses to hormone treatments in Pelibuey ewes

To determine the effects of suckling on postpartum (pp) reproductive efficiency in Pelibuey ewes, two experiments were performed. In Experiment 1, 112 ewes were randomly assigned to one of two groups at parturition: Without restriction of suckling (WRS) 24 h day(-1) for 60 days (n=56), and Weaned Ewes (WE), weaned at 40 days pp (n=56). On Day 30 pp, all ewes were given Prostaglandin (PGF2alpha) and one of four treatments (n=14): T1, intravaginal progestagen (FGA; 40 mg) for 12 days from day 30 pp+equine chorionic gonadotropin (eCG; 300 UI) until 2 days before removing FGA; T2, FGA was applied for 12 days; T3, a second application of PGF2alpha was given on day 40 pp+eCG on the same day; T4, a second injection of PGF2alpha was applied on day 40 pp only. In all the analyzed characteristics, the best results were obtained in WE. Within the WE group, the best treatment (P<0.05) was T1 with 85.7% of the ewes in oestrus, 71.4% pregnant and a prolificacy of 1.9. Within the WRS group the best results were observed in T1. In both groups, the lowest results (P<0.05) were obtained in T4. In Experiment 2, 75 ewes were randomly assigned to one of three groups (n=25) immediately after parturition: Group 1, Without restriction of suckling (WRS, as in Experiment 1); Group 2, with restriction of suckling (RS; suckling for 30 min day(-1)); Group 3, Early Weaning (EW: at 7 days pp). All ewes were given PGF2alpha at 30 days pp and the same hormonal treatment, FGA for 12 days+PGF2alpha and eCG 2 days before removing FGA. No differences were observed (P>0.05) between RS and EW for the presentation of oestrus (96% vs. 92%), pregnancy (72% vs. 76%) or prolificacy (1.9 vs. 1.9), although group WRS did not perform (P<0.05) as well as groups RS and EW for any measure of performance. In conclusion, the combination of hormonal treatment (FGA plus eCG) with weaning at 7 or 40 days pp, or restricted suckling, improves postpartum reproductive efficiency in Pelibuey ewes, demonstrating the inhibitory role of suckling on postpartum reproduction in this breed.     

Ovulation rate, embryo mortality and intrauterine growth retardation in obese swine with gene polymorphisms for leptin and melanocortin receptors

The Mediterranean Iberian pigs are obese genotypes, due to a leptin resistance syndrome related to leptin receptor gene polymorphisms. The Iberian pig is affected by a lower prolificacy when compared to lean breeds, and thus may constitute a good animal model for adverse effects of obesity in reproductive performance. The present study determined possible differences in rates of ovulation and embryo implantation and later incidence of embryo mortality and intrauterine growth retardation (IUGR) in sows of Iberian breed (n = 23) and highly-prolific lean commercial crosses (Large White × Landrace, n = 17) at two critical periods of swine pregnancy: Day 21 (just after achievement of trophoblast attachment) and Day 35 (just after completion of the transition from late embryo to early foetal stage). Two different reproductive performances were identified in the Iberian breed; 58.3% of the females had lower ovulation rates than LWxL (13.2 ± 2.3 vs 22.5 ± 1.6, P < 0.05), but 41.7% had a similar number of ovulations (18.2 ± 3.9). However, those Iberian sows having high ovulatory rates showed a high incidence of regression of corpora lutea and embryo losses between Days 21 and 35, which was not found in Iberian females with low ovulation rates; therefore, the number of viable embryos was similar in both Iberian groups (8.2 ± 1.0 and 8.4 ± 1.0) and lower that in highly-prolific LWxL (14.8 ± 1.8, P < 0.05). At Day 35, a total of 167 conceptuses (73 LWxL and 94 IB) were evaluated for IUGR. The LWxL conceptuses were longer and wider than Iberian (69.5 ± 0.1 and 64.4 ± 0.1 vs 49.9 ± 0.1 and 41.9 ± 0.1 mm, P < 0.0001). However, Iberian conceptuses were heavier than LWxL (107.4 ± 6.6 vs 68.6 ± 2.4 g) due to a lower quantity of fluids and a higher development of the placental tissues in comparison to the embryo itself. In conclusion, current study indicates a bimodal effect of obese genotypes on reproductive performances, either by lowering ovulation rate or by increased embryo losses in the first third of pregnancy.     

None of polymorphism of ovine fecundity major genes FecB and FecX was tested in goat

The polymorphism of mutation Q249R in BMPR-IB gene (FecB) and loci FecX^I, FecX^H, FecX^G, FecX^B in BMP15 gene was analyzed by forced PCR-RFLP method in 550 individuals from 6 flocks or breeds of goats with litter size varied from 1.4 to 2.7 including Boer (209), Haimen (128), second generation of Boer goat crossed with Huanghuai goat (82), Huanghuai (71), Nubi (37) and Matou (23) goat. None of mutations was detected in these goat breeds and their crossbreed. These results suggest that fecundity of goat is not linked to the same loci in BMPR-IB and BMP15 as sheep. Therefore, it is necessary to seek for other genes or loci in order to develop marker assistance selection techniques and study the prolific mechanism of the goat.     

Association Between PCR-SSCP of Bone Morphogenetic Protein 15 Gene and Prolificacy in Jining Grey Goats

The Jining Grey is a prolific local goat breed in P.R. China. Bone morphogenetic protein 15 (BMP15) gene that controls high fecundity of Inverdale, Hanna, Lacaune, Belclare, Cambridge, and Small Tailed Han ewes was studied as a candidate gene for the prolificacy of Jining Grey goats. According to the sequence of ovine BMP15 gene, six pairs of primers were designed to detect single nucleotide polymorphisms in exon 1 and exon 2 of the BMP15 gene in both high fecundity breed (Jining Grey goats) and low fecundity breeds (Boer, Liaoning Cashmere, and Inner Mongolia Cashmere goats) by single strand conformation polymorphism (SSCP). Two pairs of primers (F1/R1 and F2/R2) were used to amplify the exon 1. Four pairs of primers (F3/R3, F4/R4, F5/R5, and F6/R6) were used to amplify the exon 2. Only the products amplified by primer F5/R5 displayed polymorphism. Results indicated that two genotypes (AA and AB) were detected in prolific Jining Grey goats and only one genotype (AA) was detected in low fecundity goat breeds. In Jining Grey goats frequencies of genotypes AA and AB were 0.10 and 0.90, respectively. Sequencing revealed two point mutations (G963A and G1050C) of BMP15 gene in the AB genotype in comparison to the AA genotype. In Jining Grey goats the heterozygous AB does had 1.13 (p < 0.01) kids more than the homozygous AA does. These results preliminarily showed that the BMP15 gene is either a major gene that influences the prolificacy of Jining Grey goats or a molecular genetic marker in close linkage with such a gene.     

Prolactin Receptor as a Candidate Gene for Prolificacy of Small Tail Han Sheep

DNA polymorphism of the ovine prolactin receptor gene (PRLR) was investigated and used to study its effect on litter size in sheep. By means of PRLR gene sequence homology between sheep and human, three primer pairs were designed for polymerase chain reaction (PCR) amplification within intron 1 and exon 10 of the PRLR gene in sheep. In these parts of the gene the single nucleotide polymorphisms were detected by PCR-single strand conformation polymorphism (SSCP) in 314 Small Tail Han ewes. These poly-morphisms were used to study the associations with litter size. The results indicated that there were three genotypes (AA, AB and BB) detected by three primer pairs. For three primer pairs the frequency of allele A was 0.96, 0.79, 0.68; and the frequency of allele B was 0.04, 0.21, 0.32, respectively. The frequency of genotype AA was 0.93, 0.62, 0.51; the frequency of genotype AB was 0.06, 0.34, 0.34; the frequency of genotype BB was 0.01, 0.04, 0.15, respectively. The Small Tail Han ewes with genotype BB or AB had 0.64–0.76 or 0.44–0.54 more lambs than those with genotype AA, respectively. These results preliminarily showed that the prolactin receptor locus is either a major gene that influences the prolificacy in Small Tail Han sheep or is in close linkage with such a gene.     

Effects of human chorionic gonadotropin (hCG) and phenobarbital on the reproductive performance of fat-tailed Ghezel ewes

Increased ewe prolificacy and lambing rates, under certain conditions of sheep husbandry, may be economical and desirable. This paper reports on the effects of human chorionic gonadotropin (hCG) and phenobarbital (PB) on fertility and lambing rate in fat-tailed Ghezel sheep. One-hundred and five cyclic ewes (1.5 to 7.5 years of age) were randomly allotted to seven groups. Estrus was synchronized with two intramuscular injections of a prostaglandin F₂ analogue (PGF₂), eight days apart. Four groups were randomly assigned to the hCG and three groups to PB experiments. Twenty-four hours after the second injection of PGF₂, the ewes in four groups were injected (i.m.), either with 1 ml normal saline (control) or with 125, 250 or 500 IU hCG. Prolificacy of the ewes which conceived at first estrus increased in ewes receiving 500 IU of hCG, as compared with control group (an increase of 0.5 lamb/ewe) but fertility and lambing rate decreased in hCG-treated groups (P<0.05). Nine days after the second injection of PGF₂, ewes in the three remaining groups were orally fed empty gelatin capsules (control), 1.0 or 1.5 g of PB per day until the next estrus. Prolificacy and lambing rate increased (P<0.05) in ewes which received 1.0 g of PB (1.57 and 146.5%, respectively), as compared with the control groups (1.17 and 100.3%, respectively). It is concluded that daily feeding of 1.0 g of PB from day 7 of the estrous cycle until the next estrus period is an effective method of increasing lambing rate. hCG also increases the prolificacy, but it greatly decreases fertility and lambing rates. Since hCG is more convenient to administer on a flock basis, further experiments to improve its efficacy are warranted.