Spermatic and oxidative profile of domestic cat (Felis catus) epididymal sperm subjected to different cooling times (24, 48 and 72 hours)

Cooling stored epididymal samples for several days allows facilities to transport and process genetic material post‐mortem. Improvements to this practice allow the preservation of sperm from domestic cats, which are the ideal study model for wild felids. However, the modifications in spermatic features and the oxidative profile are not fully understood in cats. This information is necessary for the development of biotechniques, such as new extenders for cryopreservation. Therefore, the purpose of this study was to evaluate the spermatic and oxidative profile in samples from the epididymal cauda of domestic cats cooled at 5°C for 24, 48 and 72 hr. Spermatozoa were collected from the epididymis cauda. Evaluations consisted of computer‐assisted sperm analysis (CASA), plasma membrane integrity (eosin/nigrosin), acrosome integrity (fast green/rose bengal), sperm morphology, sperm DNA integrity (toluidine blue), mitochondrial activity (3′3 diaminobenzidine), activity of the antioxidant enzymes glutathione peroxidase (GPx) and superoxide dismutase (SOD), measurement of lipid peroxidation (TBARS) and protein oxidation. A decrease in sperm motility parameters was observed after 72 hr of cooling (i.e. total and progressive) with a higher percentage of minor (37.7 ± 6.3%) and total defects (53.4 ± 6.3%). Additionally, a decrease in high mitochondrial activity (Class I: 16.6 ± 2.2%) occurred after 72 hr. The decrease in motility rates after a long cooling time probably was caused by the increase in sperm abnormalities. A long cooling time causes cold shock and mitochondrial exhaustion, but there was no observed change with the oxidative stress condition. Therefore, cat epididymal sperm stored at 5°C appear to maintain a high quality for up to 48 hr of cooling time.     

A fast,low‐cost and efficient method for the diagnosis of sperm DNA fragmentation in several species

Sperm DNA fragmentation is a condition that interferes directly in the reproductive efficiency. Currently, there are several methods for assessing the sperm DNA integrity, such as Alkaline Comet, TUNEL and Sperm Chromatin Structure Assay. However, many of these techniques are laborious and require high‐precision equipment. Thus, the development of new techniques can optimize the evaluation of sperm DNA damage. Therefore, the aim of this study was to standardize the toluidine blue (TB) stain technique for the analysis of DNA fragmentation of dog, cat, bull, stallion and ram spermatozoa. For this purpose, we used six animals of each specie (n = 30), in reproductive age. Sperm was collected by different methods according to the particularities of each species, and such samples were divided into two aliquots: a sperm sample was kept at 5°C (considered as intact sperm DNA), and the remaining samples were submitted to the induction of DNA fragmentation by exposure to ultraviolet light for 4 hr. Samples were then mixed with the intact sample to obtain known and progressive proportions of sperm with fragmented DNA (0%, 25%, 50%, 75% and 100%). Semen smears were performed and subjected to staining with TB. Blue‐stained spermatozoa were considered to have DNA fragmentation. We observed high linear regression coefficients between the expected proportion of damaged DNA and the results of TB for dog, cat, ram, bull and stallion samples. In conclusion, TB stain was considered a fast and effective technique for the study of spermatozoa DNA in several species.     

Effects of five cryoprotectants on proliferation and differentiation‐related gene expression of frozen‐thawed bovine calf testicular tissue

The cryopreservation of testicular tissue is a potential method for preserving male fertility. However, the effect of cryopreservation on bovine calf testicular tissue is scarce. This study investigated the effect of different cryoprotectants on bovine calf testicular tissue at the molecular level. Testicular tissue from ten immature bovine calves (6 months) was collected after slaughter and cryopreserved in an extender containing different concentrations of the following five cryopreservation solutions (CP): bovine serum albumin (BSA) with 5% dimethyl sulfoxide (DMSO), trehalose with 5% DMSO, DMSO and glycerol and ethylene glycol (EG). After 7‐day cryopreservation, the expression levels of three spermatogonial stem cell (SSC)‐related genes, octamer‐4 (OCT4), KIT ligand (MGF/SCF) and kit oncogene (C‐KIT), were investigated by quantitative PCR (qPCR). The cell viability was highest for the tissues preserved with 30 mg/ml BSA (77.82% ± 1.22) and 40 mg/ml trehalose (74.23% ± 1.16) compared with other groups (p < 0.05), and the level of expression of the three genes was highest with 30 mg/ml BSA (p < 0.05). Compared with other CPs, the 30 mg/ml BSA and 40 mg/ml trehalose have the better cryopreserve protection. The 30 mg/ml BSA is the most viable media for the cryopreservation of testicular tissue from cattle.     

紫花苜蓿茎秆组织中木质素的分布与沉积模式

以甘农5号紫花苜蓿(Medicago sativa‘Gannong No.5’)为材料,采用组织化学染色方法研究了茎秆中木质素的发生、分布与沉积规律。结果表明,苜蓿茎秆中木质素的分布与生长部位密切相关,在茎秆顶端初生维管组织中仅木质部有木质素分布,随节间下移,木质素开始在初生韧皮纤维、次生木质部和髓射线中大量沉积;茎秆中存在G、S两种木质素,S木质素的发生迟于G木质素;在维管束之间,两种木质素均以"厚角组织处维管束→厚角组织间维管束"的模式沉积,但在维管束内,木质素沉积表现出异质性,G木质素沉积模式为"初生木质部导管→初生韧皮纤维→次生木质部和髓射线";S木质素沉积模式为"初生韧皮纤维→次生木质部和髓射线→髓薄壁细胞"。分析认为,紫花苜蓿茎秆中木质素特殊的沉积模式可能是其对北温带生长环境的一种适应对策。     

新疆乌鲁木齐市犬布鲁氏菌病流行病学调查

为了解新疆乌鲁木齐市犬布鲁氏菌病流行特征,分析犬感染风险因素,初步掌握养犬人员对布鲁氏菌病的认知程度和行为特点,从而提出合理的预防对策和控制措施,采用流行病学横断面研究方法,通过估计流行率计算抽样数量,然后随机抽样,采用IELISA方法检测光滑型和粗糙型犬布鲁氏菌,同时结合调查问卷,获得乌鲁木齐市犬布鲁氏菌病流行率、感染风险因素和养犬人员对布鲁氏菌病认知情况,并对调查结果采用SPSS和Excel软件进行数据统计分析。结果显示:乌鲁木齐市犬布鲁氏菌病总流行率为25.5%(95%CI:20.9%~30.1%),且犬种和牛羊种并存,公共卫生威胁严重;从不同群体看,牧区流行率最高,为41.5%(95%CI:31.0%~52.0%);外购、散养、常接触牛羊、喂生肉、不处理犬粪便等行为是乌鲁木齐市犬布鲁氏菌感染的潜在风险因素,其中"常接触牛羊"(OR=1.97,95%CI:1.47~15.35)、"喂生肉"(OR=5.06,95%CI:1.69~20.74)是主要风险因素;乌鲁木齐市养犬人员对布鲁氏菌病的认知程度低,个人防护意识差。因此,应加强对犬感染牛羊种布鲁氏菌的公共卫生防控,减少犬与牛羊的直接接触,不喂生肉,定期开展犬布鲁氏菌病专项检测,同时加强宣传,提高养犬人员的布鲁氏菌病认知度。     

对兽医卫生信息资源整合共享的思考

近年来,兽医卫生信息化建设蓬勃发展,信息资源愈加丰富,但存在各自为政、重复采集、一数多源、标准规范缺失、共享利用不足等问题。为打通数据壁垒,深入挖掘信息资源价值,提升兽医卫生风险防控能力,本文提出了统筹规划国家、省级两级信息平台建设,制定标准规范,加强数据共享交换和大数据分析应用的兽医卫生信息资源整合共享思路。     

亚龙湾海域生态旅游及环境保护

从地理位置、水质及沉积物、红树林、珊瑚礁海岸等方面介绍亚龙湾生态自然环境特点，分析其生态旅游发展现状、形式及成效，提出对亚龙湾生态环境保护的建议。     

椰子组培研究四十年

组织培养对于椰子快速繁殖以及全球椰子产业发展具有重要意义。本文就40年来国内外在椰子组培快繁技术,包括椰子组培技术的重要性,不同外植体如胚芽、花序、子房等在组织培养中的应用与优劣,不同培养基和培育环境对组培效果的影响等要素,进行了详细的总结和讨论。同时总结了椰子组培快繁技术的进展,分析了目前限制椰子组培快繁技术发展的因素,提出了应对策略与具体建议。本文对椰子快繁技术及椰子种质资源保存具有参考价值。     

不同固定条件对几种植物样品超微结构的影响

为比较几种固定条件对不同植物材料样品电镜超微结构图像清晰度的影响。以3个不同科属4种植物[油棕(Elaeis gineansis Jacq.)、拟南芥(Arabidopsis thaliana.)、花生(Arachis hypogaea Linn.)、柱花草(Stylosanthes guianensias Sw.)]为研究主体代表,探讨了2.5%戊二醛与4%戊二醛固定过夜、锇酸固定2 h及过夜的制备方法对其叶片超微结构的影响,并成功观察了柱花草接种胶苞炭疽野生型菌株CH008后的病菌侵入过程。结果表明:油棕、拟南芥、花生及柱花草叶片经4%戊二醛与1%锇酸固定液固定过夜,其组织成像的效果均较为理想,各种细胞器整体信息丰富、结构组织完好、线条清晰。表明,此制备方法能够较好的保存植物叶片和细胞组织,提高制样的成功率及观察分辨率。