睾丸支持细胞的功能、分离纯化及鉴定研究进展

支持细胞对维持精子形成过程中的微环境起决定作用,它可以通过分泌功能、细胞间连接形成的血睾屏障功能以及吞噬功能等来促进精子的形成过程,其发育异常会导致不同程度的雄性生殖缺陷。基于支持细胞在雄性动物生殖过程中的作用,体外培养高纯度支持细胞可成为研究睾丸两大核心功能-精子发生和性激素分泌功能相关调节机制重要的细胞模型。此外,体外培养睾丸支持细胞也可作为生殖毒理学等新兴热点领域的细胞模型,为评估和研究环境因素对雄性生殖的影响提供便利。因此,作者系统地归纳、总结了目前关于动物支持细胞生物功能的研究及常用的体外分离纯化、培养及鉴定方法,以期为利用动物支持细胞开展雄性生殖领域的研究提供参考。     

lncRNA作为竞争性内源分子的作用机制及研究进展

竞争性内源RNA （competing endogenous RNA,ceRNA）假说提出具有相同短序列非编码微小（microRNA,miRNA）应答元件（microRNA response element,MRE）的转录物通过竞争的方式结合miRNA,从而影响转录物的表达水平。ceRNA假说颠覆了miRNA与靶基因单向调控的传统观念,在RNA调控网络中具有重要的生物学意义。在众多转录物中,长链非编码RNA （long non-coding RNA,lncRNA）是一类序列长度超过200个核苷酸的非编码RNA,对lncRNA的研究涉及到遗传、分子生物、基因调控、疾病（癌症、神经系统疾病等）等领域。miRNA与lncRNA形成了一个相互作用的调控网络,lncRNA可作为ceRNA抑制miRNA的功能,从而影响后续基因的表达。近年来,随着生物信息学技术的发展,科研人员发现ceRNA作用机制不仅涉及到人类癌症疾病,而且在各种复杂动物的肌细胞分化、脂肪细胞分化和颗粒细胞凋亡等生物过程中也发挥重要的调控作用。作者追溯了ceRNA调控机制,分析了ceRNA网络调控的影响因素,综述了ceRNA在不同动物中调控miRNA的研究进展,为进一步研究lncRNA与miRNA的调控网络提供参考,为畜牧业发展及复杂动物疾病治疗提供新的思路。     

保温型刚性护面渠道设计

在刚性面层下设置一道性能良好的保温层来防止基土冻结,可以从根本上解决冻胀破坏的问题。这种带保温层的刚性护面渠道称为保温型刚性护面渠道,根据热工原理,提出了保温型刚性护面渠道横断面设计在构造方面的要求,进一步推导出计算保温层厚度的有关公式,并给出算例,还指出了需要进一步研究和探讨的问题。     

Isolation and Identification of BPIV3 Genotype C

To identify the infection agents from Ningxia Hui Autonomous region, where feedlot cattle indicated bovine respiratory disease complex (BRDC), the M gene of the bovine parainfluenza virus type 3 was amplified by RT-PCR.The PCR product was ligated to pMD18-T vector and cloned to E.coli DH5α.The positive clones were sequenced and compared with the reference strains in GenBank by the molecular biology software.Sequence alignment results showed that a BPIV3 strain was isolated from the samples and named NX49, the M gene of NX49 included 1 056 nucleotides.Evolutionary analysis showed that the NX49 belonged to BPIV3 C genotype and shared 99.4% nucleotide identity with that of the SD0835 isolated in Shandong province.The characterization of the NX49 demonstrated that it was sensitive to temperature, acid and organic matter.The presence of Mg²⁺ showed no protection against the treatment at high temperature.The HA test suggested that the NX49 enables to agglutinate the guinea pig RBC at 4 ℃ and the titer was 1∶4.The study isolated a BPIV3 genotype C strain successfully, which facilitate the study of molecular evolution and epidemiology of BPIV3 in China.     

Effects of apple pomace proportion levels on the fermentation quality of total mixed ration silage and its digestibility,preference and ruminal fermentation in beef cows

Four Japanese black beef cows were used in a 4 × 4 Latin square to evaluate the fermentation quality, digestibility, ruminal fermentation and preference of total mixed ration (TMR) silages prepared with differing proportions of apple pomace (AP). Experimental treatments were the control (no AP added, CAP), 5% (low, LAP), 10% (medium, MAP) and 20% (high, HAP) of TMR dry matter (DM) as AP. All TMR silages were well preserved. Ethanol was produced in silages containing AP and the amount increased with the proportion of AP (P < 0.05). Nutrient digestibility with LAP, MAP and HAP treatment was lower than that with CAP treatment (P < 0.05). The ruminal molar proportion of acetic acid increased (P < 0.05), but the ruminal ammonia‐N concentration decreased (P < 0.05) as the proportion of AP increased. The preference of the animals was highest for HAP, followed by MAP, CAP and LAP. This study demonstrates that decrease in nutrient digestibility might be related to the ethanol produced naturally from AP. Therefore, the proportion of AP in TMR silages should be less than 5% of dietary DM.     

ASVCP guidelines: quality assurance for portable blood glucose meter (glucometer) use in veterinary medicine

Portable blood glucose meters (PBGM, glucometers) are a convenient, cost effective, and quick means to assess patient blood glucose concentration. The number of commercially available PBGM is constantly increasing, making it challenging to determine whether certain glucometers may have benefits over others for veterinary testing. The challenge in selection of an appropriate glucometer from a quality perspective is compounded by the variety of analytic methods used to quantify glucose concentrations and disparate statistical analysis in many published studies. These guidelines were developed as part of the ASVCP QALS committee response to establish recommendations to improve the quality of testing using point‐of‐care testing (POCT) handheld and benchtop devices in veterinary medicine. They are intended for clinical pathologists and laboratory professionals to provide them with background knowledge and specific recommendations for quality assurance (QA) and quality control (QC), and to serve as a resource to assist the provision of advice to veterinarians and technicians to improve the quality of results obtained when using PBGM. These guidelines are not intended to be all‐inclusive; rather they provide a minimum standard for management of PBGM in the veterinary setting.     

Content and distribution of different phosphorus forms in broiler chickens

To understand the background value of phosphorus in chickens, the quantitative distribution of different phosphorus forms, including total phosphorus (TP), free phosphate (FP) and phospholipid (PL), in viscera, blood and bones of broiler chickens was investigated. Results showed that phosphorus contents exhibited significant differences in different parts of chickens. TP content of breast and thigh meat was over 5.0 g/kg, in which most of the phosphorus was in the form of water‐soluble phosphates. TP content in viscera was higher than that in meat, and spleen was observed to contain the highest amount of phosphorus (10.0 g/kg). In all tested organs, FP and PL contents in liver were the highest, ranging between 1207–1989 and 81–369 mg/kg respectively. TP content in chicken bone was in the range of 52 716–136 643 mg/kg, and FP content in the bone was relatively lower than that in chicken meat.     

Comparison of sampling sites and detection methods for Haemophilus parasuis

Objective To improve the isolation rate and identification procedures for Haemophilus parasuis from pig tissues. Design Thirteen sampling sites and up to three methods were used to confirm the presence of H. parasuis in pigs after experimental challenge. Procedure Colostrum‐deprived, naturally farrowed pigs were challenged intratracheally with H parasuis serovar 12 or 4. Samples taken during necropsy were either inoculated onto culture plates, processed directly for PCR or enriched prior to being processed for PCR. The recovery of H parasuis from different sampling sites and using different sampling methods was compared for each serovar. Results H parasuis was recovered from several sample sites for all serovar 12 challenged pigs, while the trachea was the only positive site for all pigs following serovar 4 challenge. The method of solid medium culture of swabs, and confirmation of the identity of cultured bacteria by PCR, resulted in 38% and 14% more positive results on a site basis for serovars 12 and 4, retrospectively, than direct PCR on the swabs. This difference was significant in the serovar 12 challenge. Conclusion Conventional culture proved to be more effective in detecting H parasuis than direct PCR or PCR on enrichment broths. For subacute (serovar 4) infections, the most successful sites for culture or direct PCR were pleural fluid, peritoneal fibrin and fluid, lung and pericardial fluid. For acute (serovar 12) infections, the best sites were lung, heart blood, affected joints and brain. The methodologies and key sampling sites identified in this study will enable improved isolation of H parasuis and aid the diagnosis of Glässer's disease.     

Characterization,phylogenetic analyses and pathogenicity of Bipolaris setariae associated with brown stripe disease on sugarcane in Yunnan,China

Brown stripe disease is a severe foliar fungal disease of sugarcane worldwide and is widespread in all sugarcane planting areas in China. Brown stripe is a major disease that seriously affects the output and quality of the sugarcane industry in Yunnan Province, China's second-largest sugar base, while the pathogen of this disease remains not yet fully understood. To address this, we isolated and identified the fungi associated with 68 leaf samples showing typical symptoms of brown stripe from 22 sugarcane varieties in different areas of Yunnan Province. A total of 113 isolates were obtained, which were morphologically similar. Of these, 64 representative isolates were sequenced for the internal transcribed spacer region (ITS), GAPDH and EF-1α loci. All representative isolates grouped with the type strain of Bipolaris setariae in the phylogenetic trees inferred with individual and concatenated sequences of ITS, GAPDH and EF1-α. Pathogenicity test results showed that B. setariae strains were able to induce typical symptoms of brown stripe. The results obtained in this study clarify that only B. setariae is associated with sugarcane brown stripe in Yunnan, China. It is recorded here for the first time as a pathogen causing sugarcane brown stripe in Yunnan, and it is able to infect many major cultivars and new varieties, posing a new threat to the sugar industry in Yunnan Province. In addition, these results provide the scientific basis for the future breeding of disease-resistant varieties and effective prevention and control of sugarcane brown stripe disease.     

二化螟dsRNA降解酶基因的克隆与表达分析

为解析二化螟Chilo suppressalis体内参与降解双链RNA(double-stranded RNA,dsRNA)的关键核酸酶的功能,克隆二化螟不同的非专一性核酸酶（non-specific nuclease,NUC）基因,并对这些基因进行生物信息学分析和组织定量表达分析,同时对dsRNA降解酶（dsRNA degrading nuclease,dsRNase）活力的组织分布进行研究。结果显示,共克隆获得5个NUC基因,其中有4个编码dsRNase亚家族基因（CsdsRNase1～CsdsRNase4）和1个编码Endonuclease G亚家族基因（CsEndoG）。5个NUC基因的开放阅读框核苷酸序列长度范围为828～1 338 bp,编码275～445个氨基酸残基,其分子量大小为31.68～49.57 kD,预测等电点为5.48～9.42。CsdsRNase1和CsdsRNase2含有信号肽序列,两者相似度极高,且与家蚕Bombyx mori和斜纹夜蛾Spodoptera litura中具有dsRNA降解酶活力的dsRNase同源聚类;CsdsRNase1和CsdsRN...