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.     

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。     

芒属植物叶绿体InDel标记的开发与应用

研究利用芒属植物(芒02381和荻04005)叶绿体全基因组测序的结果,开发12对InDel标记。采用正交设计法优化cpInDelPCR体系,得到模板DNA、dNTPs、Mg2+、引物、TaqDNA聚合酶5个关键因素在体系中的最佳组合。12对InDel标记对43份芒属和2份甘蔗材料进行扩增,共扩增出538条条带,其中3对引物扩增出13条多态性条带,占总数的2.4%。将所有位点读带转化为数值矩阵并用UPGMA法聚类,在遗传相似系数1.4的水平上,和南荻聚为一类,而芒和与五节芒、双药芒、红山茅、尼泊尔芒、甘蔗聚为一类,4个具有杂交种特征的材料单独聚为一类。试验开发的芒属植物cpInDel标记可用于芒与荻的区分,指导芒属植物的杂交育种。     

“互联网+农业”模式下食用菌的营销策略

在"互联网+农业"融合发展背景下,食用菌企业需要借助互联网,构建市场主导、数字驱动的全新营销战略,助力产品营销。以"互联网+农业"模式为研究视角,分析食用菌产品营销现状,提出发挥技术优势、挖掘用户需求和建设优质平台等营销建议,以助力食用菌产品市场营销与效益转化。     

罹患口腔炎乌苏里蝮的相关生理学指标分析

口腔炎是人工养殖蛇类的常见病之一,为了深入了解和尽早做好预防工作,对罹患口腔炎的成年雄蛇乌苏里蝮(Gloydius ussuriensis)进行解剖观察,并检测了相关器官的系数、血液学指标和激素等。结果表明:整体肥满度、躯体肥满度、肾系数等数值罹患口腔炎蛇略高于健康蛇;而肝系数、精巢系数和脂肪体系数等罹患口腔炎的蛇低于健康雄蛇,且前两者呈显著性差异,后者呈极显著性差异。性激素检测结果显示罹患口腔炎的蛇血液睾酮和雌二醇含量均低于健康蛇,且均具极显著性差异。血细胞指标中,红细胞数量、红细胞分布均差、白细胞总数、淋巴细胞数量和淋巴细胞比率等均为罹患口腔炎的蛇高于健康雄蛇,且具显著性差异,但嗜中性粒细胞和淋巴细胞的比值(NLR)、中性粒细胞数量和比率、中间细胞数量及其比率均为患口腔炎的雄蛇低于健康蛇,且具显著性差异。结论:口腔炎不仅影响了主要代谢器官的器官系数的变化,而且血液多项指标和性激素水平也有明显差异。性激素含量的变化将对乌苏里蝮的正常生殖繁育产生不可忽视的影响。另外,NLR和红细胞分布宽度2个血常规指标可作为早期诊断炎症发生的首选参数。     

房地产销售系统的软件设计与实现

房地产业的飞速发展,给房地产销售管理带来了很多新的问题和任务,传统的管理模式已经不能适应现代房地产销售管理的需要,亟待建立一套房地产销售管理信息系统,以满足房地产管理信息化、科学化、现代化的需求。     

支管射流三通结构优化与试验

为了提高支管射流三通水力性能,改善滴灌的灌水均匀性,基于CFX数值模拟技术,对进口宽度为15 mm的支管射流三通进行结构优化.选取位差、劈距、劈尖半径和侧壁倾角为影响因素,通过四因素三水平正交设计了9组模型,边界条件设定为进口压力100 kPa.选取支管射流三通出口设计流量为评价标准,支管射流三通最优结构尺寸为位差5.5 mm、劈距113 mm、劈尖半径13 mm、侧壁倾角10°.此结构尺寸参数下的支管射流三通水力性能试验结果表明:在进口水压为100 kPa时,支管射流三通脉冲频率为148次/min,水头压力振幅为37.9 kPa,水头压力损失为16.7 kPa,出口流量为0.698 L/s;支管射流三通所接滴灌带长度为60 m时,与普通支管三通相比,支管射流三通的灌水均匀系数提高了2.78%,流量偏差率降低了4.72%.该研究可为射流技术在脉冲滴灌系统的研究、开发与应用提供理论依据.