An efficient method for the sanitary vitrification of bovine oocytes in straws

Background：At present,vitrification has been widely applied to humans,mice and farm animals.To improve the efficiency of vitrification in straw,bovine oocytes were used to test a new two-step vitrification method in this study.Results：When in vitro matured oocytes were exposed to 20%ethylene glycol（EG20） for 5 min and 40%ethylene glycol（EG40） for 30 s,followed by treatment with 30%glycerol（Gly30）,Gly40 or Gly50,a volume expansion was observed in Gly30 and Gly40 but not Gly50.This indicates that the intracellular osmotic pressure after a 30 s differs between EG40 and ranged between Gly40（approximately 5.6 mol/L） and Gly50（approximately 7.0 mol/L）.Since oocytes are in EG40 just for only a short period of time（30 s） and at a lower temperature（4℃）,we hypothesize that the main function of this step in to induce dehydration.Based on these results,we omitted the EG40 step,before oocytes were pretreated in EG20 for 5 min,exposed to pre-cooled（4℃） Gly50,for 30 s,and then dipped into liquid nitrogen.After warming,81.1%of the oocytes survived,and the surviving oocytes developed into cleavage stage embryos（63.5%） or blastocysts（20.0%） after parthenogenetic activation.Conclusions：These results demonstrate that in a two-step vitrification procedure,the permeability effect in the second step is not necessary.It is possible that the second step is only required to provide adequate osmotic pressure to condense the intracellular concentration of CPAs to a level required for successful vitrification.     

应用防虫网和黄板防治大棚辣椒秋延后栽培主要病虫害试验

为研究大棚秋延后辣椒主要病虫害无公害绿色防控技术,选择不同目数的防虫网与黄板配套应用,以本地传统化学农药防治方法为对照,调查烟粉虱、蚜虫和病毒病的为害情况,并统计大棚辣椒产量。结果表明,应用60目防虫网和黄板的防治效果较使用化学农药的好,能有效防治大棚烟粉虱、蚜虫和病毒病,操作简便,增产明显,值得在大棚蔬菜生产中推广应用。     

对虾病毒HHNBV DNA构建质粒的研究与分析

用ＰＵＣ１８构建了对虾病毒ＨＨＮＢＶＤＮＡ重组质粒，提取后经斑点杂交及酶切分析，证实质粒中插入片段为病毒ＤＮＡ，其中０５＃质粒的插入片段大于２Ｋｂ，与质粒的分子量相近。对０５＃质粒酶切后的Ｓｏｕｔｈｅｒｎ杂交亦取得了满意的结果。同时，还对０５＃质粒的插入片段进行了内切酶图谱分析，共进行了ＥｃｏＲｌ、Ｈｉｎｄｌｌｌ、Ｓｍａｌｌ、Ｐｓｔｌ、ＰＵＶｌ、Ｈｉｎｄｌｌ６种限制性内切酶分析，结果表明，只有Ｐｓｔｌ在插入片段的一端有一个酶切位点，酶切位点的确切位置有待于进一步确定。     

观赏鱼类疾病及其诊断方法

观赏鱼因经长期驯化变种而来比较娇贵,且所处环境也相对稳定,一旦管理不善很容易引起疾病。本文主要讨论了观赏鱼饲养过程中常见疾病的种类、发病原因、诊断方法等,重点介绍了:免疫学诊断技术、核酸诊断技术、核酸技术与免疫学相结合的方法。免疫学诊断技术包括单克隆抗体技术、荧光抗体技术等。核酸诊断技术包括核酸杂交技术、PCR(聚合酶链式反应)技术、16S rRNA检测技术。核酸技术与免疫学相结合的方法有PCR-ELISA法等。     

Enteropathogenic bacteria in dogs and cats: diagnosis, epidemiology, treatment, and control

This report offers a consensus opinion on the diagnosis, epidemiology, treatment, and control of the primary enteropathogenic bacteria in dogs and cats, with an emphasis on Clostridium difficile, Clostridium perfringens, Campylobacter spp., Salmonella spp., and Escherichia coli associated with granulomatous colitis in Boxers. Veterinarians are challenged when attempting to diagnose animals with suspected bacterial-associated diarrhea because well-scrutinized practice guidelines that provide objective recommendations for implementing fecal testing are lacking. This problem is compounded by similar isolation rates for putative bacterial enteropathogens in animals with and without diarrhea, and by the lack of consensus among veterinary diagnostic laboratories as to which diagnostic assays should be utilized. Most bacterial enteropathogens are associated with self-limiting diarrhea, and injudicious administration of antimicrobials could be more harmful than beneficial. Salmonella and Campylobacter are well-documented zoonoses, but antimicrobial administration is not routinely advocated in uncomplicated cases and supportive therapy is recommended. Basic practices of isolation, use of appropriate protective equipment, and proper cleaning and disinfection are the mainstays of control. Handwashing with soap and water is preferred over use of alcohol-based hand sanitizers because spores of C. difficile and C. perfringens are alcohol-resistant, but susceptible to bleach (1:10 to 1:20 dilution of regular household bleach) and accelerated hydrogen peroxide. The implementation of practice guidelines in combination with the integration of validated molecular-based testing and conventional testing is pivotal if we are to optimize the identification and management of enteropathogenic bacteria in dogs and cats.     

牛肌肉生长抑制素基因的研究进展

肌肉生长抑制素,属TGF-β超家族,是一种骨骼肌生长发育的负调控因子。其生物活性的丧失或受到抑制,会导致骨骼肌增生从而引起动物肌肉的过度发育,表现为双肌性状。本文综述了MSTN基因的发现、结构特点、作用机制、在牛科物种中的研究进展及其应用前景。     

益生素对断奶仔猪生产性能和腹泻率的影响

本试验选用35±2日龄的PIC五元杂交配套系断奶仔猪45头,将其随机分成3个试验组,即试验1组、试验2组和对照组,在试验组仔猪日粮中分别添加0.1%和0.3%的复合益生菌制剂,旨在研究益生素对断奶仔猪生产性能的影响。试验结果表明,益生素对断奶仔猪的生产性能有良好的促进作用,且腹泻率有所降低。     

Reactivity and prevalence of neutralizing antibodies against Japanese strains of bovine viral diarrhea virus subgenotypes

Bovine viral diarrhea virus (BVDV) field isolates show genetic and antigenic diversity. At least 14 subgenotypes of BVDV-1 and 4 of BVDV-2 have been identified in Artiodactyla worldwide. Of these, 6 subgenotypes of BVDV-1 and 1 of BVDV-2 have been isolated in Japan. Previously, we reported that each subgenotype virus expresses different antigenic characteristics. Here we investigated the reactivity of neutralizing antibodies against representative strains of Japanese BVDV subgenotypes using sera from 266 beef cattle to estimate the prevalence of this epidemic virus among cattle in Japan. Antibody titers at concentrations at least 4-fold higher than antibodies against other subgenotype viruses were considered subgenotype specific. Subgenotype-specific antibodies were detected from 117 (80.7%) of 145 sera samples (69.7% against BVDV-1a, 1.4% against BVDV-1b, 8.3% against BVDV-1c, and 1.4% against BVDV-2a). The results suggest that neutralization tests are useful in estimating currently epidemic subgenotypes of BVDV in the field.     

Evaluation of osteoinduction properties of the demineralized bovine foetal growth plate powder as a new xenogenic biomaterial in rat

The aim of this study was evaluation of osteoinductive properties of demineralized bovine foetal growth plate in submuscular transplantation (ectopic osteoinduction) as a new xenogenic biomaterial in rat model. Demineralized bovine foetal growth plate was ectopically implanted in 18 male Sprague–Dawley rats. In 18 of the animals under aseptic conditions two submuscular pouches were created between external and internal oblique abdominal muscles in the two flanks: the right was left empty (sham) and the left was filled with 20 mg of demineralized bovine foetal growth plate powder. Radiographs were taken in 2, 4 and 6 weeks after the surgery, then six animals were pharmacologically euthanized after 2, 4 and 6 weeks for histopathological evaluation. Results showed: (1) osteoinductivity of xenogenic demineralized bovine foetal growth plate powder, and (2) earlier mineralization of ectopically implanted demineralized bovine foetal growth plate in the submuscular implanted area. Our results show that submuscular implantation of xenogenic demineralized bovine foetal growth plate has osteoinductive properties in a rat model.     

History and selection imprinting on genetic relationships among bovine breeds analyzed trough five genes related with marbling

Many candidate genes have been suggested as responsible for marbling in beef cattle, for instance diacylglycerol O-acyltransferase 1, thyroglobulin, growth hormone, leptin and stearoyl CoA desaturase. The objective of the present work was to evaluate the polymorphisms of five SNPs of these candidate genes in 389 animals of 18 Bos Taurus and Bos indicus breeds. The obtained results were compared with ones previously obtained with STRs and loci related to milk production in these populations. Moreover we analyzed whether the phylogenies reconstructed using SNPs associated with marbling resulted in the known tree topology. The tree constructed with UPGMA, using genetic distance D_A, exhibit a topology partially consistent with the historical origin of breeds. The result observed in the Correspondence Analysis coincided with the topology of the UPGMA tree. This work allowed us to evaluate the five SNPs genetic diversity and to demonstrate that the grouping of the breeds may be the result of its history, selection process, or both at once.