Mapping genomic regions for reproductive traits in beef cattle: Inclusion of the X chromosome

Although the second largest chromosome of the genome, the X chromosome is usually excluded from genome-wide association studies (GWAS). Considering the presence and importance of genes on this chromosome that are involved in reproduction, the aim of this study was to evaluate the effect of its inclusion in GWAS on reproductive traits (scrotal circumference [SC], early pregnancy [P16] and age at first calving [AFC]) in a Nelore herd. Genotype data from 3,263 animals with the above-mentioned phenotypes were used. The results showed an increase in the variances explained by the autosomal markers for all traits when the X chromosome was not included. For SC, there was an increase of more than 10% for the windows on chromosomes 2 and 6. For P16, the effect was increased by almost 20% for windows on chromosome 5. The same pattern was found for AFC, with an increase of more than 10% for the most important windows. The results indicate that the noninclusion of the X chromosome can overestimate the effects of autosomes on SC, P16 and AFC not only because of the additive effect of the X chromosome itself but also because of its epistatic effect on autosomal genes.     

Determination of nucleotide sequence of SRY gene in sika deer (Cervus nippon)

The aim of the present study was to determine the whole nucleotide sequence of the open reading frame of the sex‐determining region Y (SRY‐ORF) in wild sika deer. The SRY gene of wild sika deer was obtained by polymerase chain reaction (PCR) with DNA from blood samples. The whole nucleotide sequence of the SRY‐ORF in wild sika deer consisted of 687 bp and encoded 229 deduced amino acids. In comparison with the bovine SRY gene, the percentage of nucleotide sequence homology was 91.0% in the overall ORF, and those of the N‐terminal, high mobility group (HMG) box, and C‐terminal regions within ORF were 88.9%, 96.2% and 87.9%, respectively. The nucleotide sequences of sika deer SRY‐ORF characterized in the present study can be used for phylogenetic analysis or sexing in wild sika deer.     

On ancestors of dog breeds with focus on Weimaraner hunting dogs

Paternally inherited Y chromosomal markers and maternally inherited mitochondrial (mt) DNA sequences were investigated in 27 dog breeds (Canis familiaris), of which the Weimaraner hunting dog was studied in greater detail. Altogether, nine potentially polymorphic markers of the Y chromosome were examined as well as parts of the canine mt genome (1947 base pairs) in 111 male dogs and four wolves for comparison. Twenty Y chromosomal and fifty-nine mitochondrial DNA (mtDNA) haplotypes were identified in the canine breeds and wolves. In 34 Weimaraners, four distinct Y chromosomal haplotypes were observed as well as three mtDNA types thus reflecting at least four male and three female ancestors for the current population in Germany. Tracing patri- and matrilineages, several entries in the Weimaraner stud book cannot be reconciled with the male-only, Y chromosomal neither the female-only, mt inheritance patterns, respectively. The investigated breeds represent 9 of 10 groups defined by the Fédération Cynologique Internationale (FCI). The level of Y chromosomal and especially mtDNA diversity was immense considering the relatively small number of individuals investigated per breed. Unique haplotypes were found only in a few breeds and the wolf. Other haplotypes were shared among several breeds, also across different FCI groups, suggesting that these canine breeds had common male and female ancestors.     

Competition Horses Housed in Single Stalls (I): Behavior and Activity Patterns during Free Exercise According to Its Configuration

Although welfare of animals has become more important, housing horses in systems that limit natural behavior patterns is still widespread. Single stalls are the predominant housing system in Germany, especially for competition horses. Free exercise on fields or paddocks helps to improve welfare for the animals, but allowing it is not taken for granted by many horse keepers. In the present study, three common management practices were investigated with regard to their effect on the behavior of six competition horses housed in single stalls: 2-hour turnout after training in groups of two horses (group turnout [GT]), solitary turnout (ST) after training for 2 hours, and no turnout (NT) in addition to training. The aim of the study was to determine the differences in the horses’ behavior during turnout, training, and in the stable caused by the treatments. This article focuses on the behavior and activity patterns during turnout. The horses’ behavior was observed directly by the scan sampling method (5-minute interval), and the distance covered during turnout was measured by global positioning system devices. During turnout, the horses spent most of the time eating. After 1 hour of being turned out, eating decreased, and occupation, locomotion, and social interaction increased. When turnout was allowed in groups (GT), the horses spent more time in social interactions and less time walking and trotting than in ST. Therefore, they also covered a greater distance in ST than in GT. The distance covered in ST also showed a greater deviation both within and between days compared with GT.     

Exploring polymorphisms and their effects on reproductive traits of the INHA and INHβA genes in three goat breeds

In this study, we report the analysis of INHA and INHβA gene polymorphisms in 786 goats of three breeds: Xinong Saanen (SN), Guanzhong (GZ) and Boer (BG). We identified three new allelic variants: P1–C80G and/126G (GenBank accession no. HQ202573) in the three goat breeds and P2–C936T (GenBank accession no. HQ202572) in SN and GZ goat breeds. At P1 locus, AA, AB and BB genotypes were found in the three goat breeds. At P2 locus, CC and CT genotypes were found in SN and GZ goat breeds. After comparing genotype distribution within the three goat breeds, BG had conspicuous differences from SN and GZ (P < 0.001) at P2 locus. The SNP locus was in Hardy–Weinberg disequilibrium at P1 locus in the three goat breeds (P < 0.05). At P2 locus, the SNP locus was in Hardy–Weinberg disequilibrium in SN and GZ goat breeds (P < 0.05). Association of polymorphisms with litter size was done at P1 locus in the three goat breeds. The result showed that AA genotype had remarkable litter size at P1 locus in the three goat breeds (P < 0.05). Therefore, these results suggest that INHA gene is a strong candidate gene that affects litter size in goats.     

京海黄鸡IGFBP-1基因遗传多态性及其与生长性状的关联分析

以京海黄鸡、AA鸡、尤溪麻鸡、边鸡等4个鸡品种为研究对象,采用PCR-SSCP方法检测胰岛素样生长因子结合蛋白1(insulin-like growth factor-binding protein-1,IGFBP-1)第4外显子的多态性,并分析其对京海黄鸡生长性能的遗传效应。结果显示,在IGFBP-1第4外显子区检测到2处变异(5550T→C、5692AAT插入)。对于P3扩增片段,在尤溪麻鸡群体中检测到AA、AB、BB 3种基因型,在AA鸡和边鸡品种中检测到AA和AB 2种基因型,京海黄鸡中只检测到AA基因型;对于P4扩增片段,在4个鸡品种中均检测到CC、CD、DD 3种基因型。最小二乘分析结果表明,京海黄鸡3种基因型个体的初生重存在显著差异(P＜0.05)。     

Stallion spermatozoa selected by single layer centrifugation are capable of fertilization after storage for up to 96?h at 6°C prior to artificial insemination

Background

One of the challenges faced by equine breeders is ensuring delivery of good quality semen doses for artificial insemination when the mare is due to ovulate. Single Layer Centrifugation (SLC) has been shown to select morphologically normal spermatozoa with intact chromatin and good progressive motility from the rest of the ejaculate, and to prolong the life of these selected spermatozoa in vitro. The objective of the present study was a proof of concept, to determine whether fertilizing ability was retained in SLC-selected spermatozoa during prolonged storage.

Findings

Sixteen mares were inseminated with SLC-selected sperm doses that had been cooled and stored at 6°C for 48 h, 72 h or 96 h. Embryos were identified in 11 mares by ultrasound examination 16–18 days after presumed ovulation.

Conclusion

SLC-selected stallion spermatozoa stored for up to 96 h are capable of fertilization.     

磷酸二酯酶在猪卵巢的表达与相关中药对其活性的影响

为了测定猪卵巢组织磷酸二酯酶（PDE）同工酶基因表达类型与活性规律，研究相关中药的作用机理，为繁殖障碍性疾病治疗积累资料。作者提取中国实验用小型猪卵巢组织总RNA，应用反转录聚合酶链反应（RT-PCR）测定PDE同工酶在其中的表达。选取催情散等复方及单味药，运用高效液相色谱法（HPLC）根据作用前后cAMP/cGMP含量的变化，分析对PDE活性的影响。结果检测的18个PDE亚型中除PDE4C外，PDE 1A、1B、1C、2A、3A、3B、4A、4B、4D、5A、7A、7B、8A、8B、9A、10A、11A mRNA在卵巢组织中均有表达。卵巢组织PDE活性较强，其中cGMP-PDE活性大于cAMP-PDE活性。催情散对卵巢组织cGMP-PDE活性均有较强的抑制作用，其中单味药淫阳藿作用最强。研究结果表明卵巢组织含有丰富的PDE同工酶，在通过环核苷酸水平卵巢机能调节中发挥着重要作用，中药催情散的作用与显著抑制cGMP-PDE活性有关。     

鸭生长激素(GH)基因编码区及调控区多态性分析

根据鸭生长激素基因编码区及调控区的序列设计8对引物,利用PCR-SSCP方法对北京鸭、西湖野鸭、樱桃谷鸭、金定鸭、山麻鸭、荆江鸭、绍兴鸭、缙云麻鸭等8个鸭种进行单核苷酸多态性分析。结果共发现3个突变位点,分别为230处（C→G）、244处（C→A）和3 701处（C→T）。前两处突变位于5′调控区,3 701处突变位于编码区第4外显子,但该编码区的突变是沉默突变,3′调控区表现了高度的保守性。统计结果发现：⑴在5′调控区基因座上,金定鸭的等位基因B频率显著高于其他品种;⑵在外显子4基因座上,基因型频率的分布与品种有关,且肉用型鸭的CC基因型频率显著高于蛋用型。可以推测,本研究所检测到的基因座可能与生产性能相关。     

用响应面分析法优化桑叶水溶性多糖的提取工艺

为了高效提取桑叶中的水溶性多糖,通过单因素试验考察了提取温度、提取时间、原料质量浓度和提取次数对多糖得率的影响,然后采用响应面分析法优化桑叶水溶性多糖的提取工艺。以提取温度和原料质量浓度为主要因素进行中心组合试验,建立了多糖得率测算模型的回归方程,确认提取温度和原料质量浓度对多糖得率都有显著影响。优化后的提取工艺在温度90℃、提取时间80 min、原料质量浓度0.042 g/mL的条件下提取1次,桑叶水溶性多糖的最高得率达到1.92%。