日粮鱼油对高脂日粮饲喂小鼠发情周期和机体产热的影响

本研究旨在探讨鱼油对高脂日粮饲喂小鼠发情周期和机体代谢产热的影响。试验选用36只4周龄C57BL/6 J雌性小鼠,随机分成3组（n=12）：对照组、高脂组和高脂+鱼油组。对照组饲喂标准啮齿动物饲料（AIN-93G）,高脂组和高脂+鱼油组分别饲喂高脂日粮（脂肪提供60%能量）和添加5%鱼油（等能替代猪油）的高脂日粮。试验期间,对小鼠体组成（12周龄）、整体代谢（16周龄）、褐色脂肪温度（18周龄）、体核温度（直肠温度,18周龄）和发情周期（20周龄）等进行检测。试验结束后,眼球采血分离血清,检测促卵泡激素（follicle-stimulating hormone,FSH）和雌二醇（estradiol,E2）的水平。此外,采集皮下脂肪、腹部脂肪和肩胛间褐色脂肪,称重并使用Western blot检测脂肪组织中产热相关基因的蛋白表达（UCP1、Cyto C）,使用实时荧光定量PCR检测褐色脂肪组织中产热基因的mRNA表达（UCP1, PRDM16,PGC1α,Cidea,Elovl3）。结果显示,与对照组相比,高脂日粮显著增加了小鼠的体脂含量（12周龄）及皮下和腹部脂肪的沉积量（21周龄）（P<0.05）,而添加鱼油显著降低了高脂饮食引起的体脂含量增加（P<0.05）。另外,高脂日粮导致小鼠的发情周期紊乱,伴随着周期延长、发情期缩短,以及血清中FSH和E2的水平降低（P<0.05）,而添加鱼油可缓解高脂日粮导致的小鼠发情周期紊乱,提高血清中FSH和E2的水平（P<0.05）。同时,添加鱼油可增加高脂饲喂小鼠肩胛间褐色脂肪（interscapular brown adipose tissue,iBAT）和腹股沟白色脂肪（inguinal white adipose tissue,iWAT）中产热相关基因的表达（P<0.05）,进而促进iBAT激活/产热和iWAT褐色化。结果提示,日粮鱼油可缓解高脂日粮导致的发情周期紊乱,可能与BAT激活和WAT褐色化造成的机体代谢产热增强有关。     

长白和大白猪主要生长性状的遗传参数估计

旨在分析母猪的出生年份、出生季节、初生重、开测日龄等固定效应对长白、大白猪主要生长性状的影响,并对目标生长性状进行遗传参数估计（遗传力、遗传方差、表型相关和遗传相关）,为猪的遗传改良提供基本依据。本试验利用GLM模型分析试验猪群（398头长白猪和1 176头大白猪）的固定效应对猪生长性状的影响,并采用多性状动物模型对目标性状进行遗传参数估计。目标生长性状包括达100 kg体重日龄（age to 100 kg,AGE）、达100 kg背膘厚（backfat to 100 kg,BF）、100 kg平均日增重（average daily gain to 100 kg,ADG）。研究表明,在大白和长白猪中,猪的出生年、出生季、初生重以及开测日龄对生长性状均具有极显著的影响（P<0.001）;长白猪的AGE、ADG和BF的遗传力分别为0.321、0.327和0.324,大白猪对应性状的遗传力分别为0.454、0.469和0.408;长白猪的ADG和AGE之间的遗传相关、表型相关分别为-0.990、-0.995,大白猪的ADG和AGE之间的遗传相关、表型相关分别为-0.993、-0.998,均呈现较强的负相关。长白、大白猪的生长性状（AGE、ADG、BF）均属于中等遗传力性状,其出生年份、出生季节、初生重和开测日龄对猪的生长性状影响较大。在遗传参数估计分析时,提高样本数量并提升表型数据质量,可以增加遗传参数估计的可靠性。本研究中的生长性状遗传参数估计结果较为可靠,可为后续的遗传改良提供参考。     

一株乳源松鼠葡萄球菌ExhC基因的克隆及序列分析

松鼠葡萄球菌(Staphylococcus sciuri,S.sciuri)是引起奶牛乳房炎(Cow Mastitis)的病原菌之一,其主要致病因子是该菌产生的脱皮毒素C(Exfoliative Toxin C,ExhC)。为了深入了解该基因的分子特征,本试验对从乳房炎患牛牛乳中分离的一株松鼠葡萄球菌ExhC基因(Genbank登录号:MT845354)进行了克隆及序列分析。按GenBank收录的ExhC基因设计并合成引物,利用PCR对ExhC基因进行扩增,并对扩增后的ExhC基因进行测序及分析。结果表明,松鼠葡萄球菌ExhC基因的开放阅读框序列长度为837bp,共编码278个氨基酸。通过Blast模块对ExhC基因进行同源性分析,得到5株相似序列,与ExhC基因序列相比同源性均为99.04%。系统进化树图指示该基因与其余五株相似序列属于不同的分支,遗传距离较远。利用TMHMM对松鼠葡萄球菌ExhC基因837bp序列跨膜区进行预测,结果显示ExhC蛋白的第1~277位氨基酸在细胞膜外,5~25位氨基酸所在位置为跨膜区。通过ExhC氨基酸序列分析和ExhC蛋白二级结构、三级结构预测,推断第97位氨基酸插入Asn以及其他位点氨基酸的突变可能会导致松鼠葡萄球菌功能区变化和毒株毒力的相对变化,这将给松鼠葡萄球菌致病机理的研究奠定基础。     

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。     

湖南长沙记录到绿背姬鹟

2019年10月3日,在湖南省长沙市烈士公园(112°59′55.74″E,28°12′48.08″N)内一池塘周边的阔叶林中观察并拍摄到一种鹟科鸟类。经查阅文献,对比该鸟的形态特征后,鉴定为绿背姬鹟(Ficedula elisae),为湖南省新纪录鸟种。     

宁夏黄灌区退水过程中氮污染特征模拟

宁夏黄灌区是我国商品粮生产基地之一,农业高强度和化肥高投入对灌区水环境和黄河水安全造成了严重威胁。农田退水中氮素迁移流失的网室土柱模拟实验表明:1每次施肥1~2d后(穗肥是1周后),田面水中全氮量均出现峰值,不断下降后逐渐趋于稳定,全氮量随时间的变化符合对数衰减规律;23次施肥后,种植水稻的土柱各土层渗漏水中的全氮量高于未种植水稻的土柱,水稻根系的发育,一定程度上促进了氮素向下迁移,加剧了氮素流失的风险,水稻收获前控制氮的渗漏损失尤为重要。     

不同灌溉方式下冬小麦穗部性状与产量关系的试验研究

采用田间试验,以豫麦69为试验材料,研究了不同水分处理下常规灌溉和一体化垄作沟灌的冬小麦产量与穗部性状的相互关系。结果表明,同常规灌溉方式相比,一体化垄作沟灌方式下,冬小麦的穗粒数、籽粒质量及产量分别增加了5.5356%、7.5489%、7.7454%,但穗数减少了0.4302%。常规灌溉和一体化垄作沟灌方式下,产量与穗数、穗粒数以及籽粒质量均正相关,但一体化垄作沟灌方式下的相关系数较常规灌溉方式大。一体化垄作灌溉有利于改善农田小气候、发挥作物的边行优势和提高小麦产量。     

Construction and Identification of Yeast Surface Display Libraries of Ovis aries DRA Gene Exon 2

The specific primers were designed according to Ovis aries DRA gene sequence deposited in GenBank and the multiple cloning site of the plasmid pYD1,which was a vector used for protein surface display on Saccharomyces cerevisiae.The gene encoding DRA was amplified by PCR using the genomic RNA of Ovis aries.The 762 bp fragment was cloned and released in GenBank and registration number was KR422362.The PCR product was inserted into the yeast surface display plasmid vector pYD1 by double enzyme digestion.It was indicated that DRA gene was successfully integrated into the genome.Dot mutation was made at both ends of exon 2 in DRA gene for making restriction enzyme cutting site and design the exon 2 specific primers according to mutated Ovis aries DRA gene sequence.Sequenced exon 2 amplification products based on DNA pooling of sheep large sample template was analyzed the polymorphic loci.The polymorphic exon 2 246 bp fragment was obtained by double enzyme digestion and connected to surface display restructuring mutation carriers pYD1-DRA by the same double enzyme digestion,and then we successfully constructed yeast surface display libraries.We transformed it into Saccharomyces cerevisiae EBY100 cell.Yeast monoclone was identified by PCR amplification and sequencing,and we confirmed that DRA gene had been integrated into Saccharomyces cerevisiae genome.After galactose induced,it was detected that DRA gene library had been successfully demonstrated on the yeast cell surface under the fluorescence microscope by immunofluorescence method.     

Screening of Probiotics Inhibiting E. coli K88 in vitro

The probiotics tested in the experiment were isolated from the intestinal of weaning piglets.The isolated probiotics and E.coli K88 were inoculated into the culture of intestinal porcine epithelial cell-1 (IPEC-1).The activity of lactate dehydrogenase (LDH) in the supernatant was measured after incubating for 2.5 h.At the same time, the probiotics and E.coli K88 were co-cultured in vitro, the number of E.coli K88 was counted and the probiotics which could be resistant to the E.coli K88 were selected 2.5 h later.The results showed that the Lactobacillus casei and Bacillus coagulans could significantly reduce LDH activity (P<0.05), decrease the damage of E.coli K88; Bacillus coagulans could inhibit the growth of E.coli K88.At the same time, Bacillus coagulans could resist high temperature, acid and bile salt.The results showed that Bacillus coagulans strains had great potential as the application of probiotics strains.The methods could be used as a model of screen probiotics which could inhibit the growth of E.coli K88 in vitro.     

Establishment of a Duplex Real-time RT-PCR Assay for the Differential Detection of Nipha Virus and Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus

To establish a rapid,sensitive and specific assay for the differential detection of Nipah virus (NiV) and highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV),a duplex Real-time RT-PCR was developed with specific primers and probes targeting to the special sequences of NiV M gene and HP-PRRSV nsp2 gene by optimization of reaction conditions.The performance of the assay was linear ranging from 4.6×10¹ to 4.6×10⁷ copies/μL for RNA standard control of NiV M (NiV-M-RNA) and from 4.1×10¹ to 4.1×10⁸ copies/μL for RNA standard control of HP-PRRSV nsp2 (HP-PRRSV-nsp2-RNA),and detection limits of the assay was 46 copies for the NiV-M-RNA and 4.1 copies for the HP-PRRSV-nsp2-RNA,respectively.The coefficients of variation (CVs) of both inter-assay and intra-assay repeatability were less than 2.0%,showing good repeatability.The assay was able to specifically detect NiV and HP-PRRSV simultaneously without cross-reaction with classical swine fever virus (CSFV),porcine epidemic diarrhea virus (PEDV),swine influenza virus (SIV),porcine parvovirus (PPV),pseudorabies virus (PRV) and porcine circovirus type 2 (PCV2).Of the 236 samples from pigs for both NiV and HP-PRRSV detection by the established assay,all the samples were negative for NiV,8 samples were HP-PRRSV positive.In conclusion,this assay offers a useful approach for the differential detection of NiV and HP-PRRSV in clinical specimens from the pigs.