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.     

Sperm subpopulations influence the pregnancy rates in cattle

This study aimed to study the characteristics and subpopulations of spermatozoa from bulls with low and high reproductive performance based on pregnancy rates. Based on historical records of pregnancy rate from four farms, 24 bulls were selected. Two groups were established, with low pregnancy rates (n = 12; LOW), including bulls that presented pregnancy rates <52.27% (33.33% to 51.81%); and a group with high pregnancy rates (n = 12; HIGH), with pregnancy rates >52.27% (52.27% to 69.64%), after fixed-time artificial insemination (FTAI). The thawed sperm straws were analysed to sperm kinetics, morphology, plasma membrane integrity and sperm subpopulations. The LOW group exhibited a higher proportion of static cells (p < .05). In contrast, the HIGH group showed greater percentages for membrane integrity and total and progressive motility, and cells with fast and medium velocity (p < .05). In the cluster procedures, four sperm subpopulations were established. The low-fertility bulls presented the highest percentage of subpopulation 2 (41.46%), characterized by slow and progressive spermatozoa. The high-fertility bulls exhibited the highest percentage of subpopulation 3 (37.17%), characterized by fast and nonlinear spermatozoa. Results from this study indicated that bulls with greater percentages of fast and nonlinear spermatozoa seem to have greater fertilization capacity and the subpopulations analysis can be considered a tool to identify ejaculates with high fertility.     

The influence of crossbreeding on collagen solubility and tenderness of Infraspinatus and Semimembranosus muscles of semi‐intensively reared young bulls

The aim of the present study was to investigate the effects of crossbreeding on collagen content and solubility, shear force (WBSF) and the eating quality of Infraspinatus (INF) and Semimembranosus (SM) muscles of young bulls and the relationships between collagen content and solubility, shear force and the eating quality of beef. The experimental material comprised muscles of crossbred young bulls (about 600 days old) of Polish Holstein‐Friesian (PHF) × Limousine (LM) (n = 10), PHF × Charolaise (CH) (n = 9), PHF × Hereford (HER) (n = 9) breeds. The crossbreeding influenced WBSF, aroma and taste, total, water‐soluble, acid‐soluble, total soluble and insoluble collagen content, as well as the acid‐soluble, total soluble and insoluble collagen proportions. WBSF was significantly negatively correlated with sensorial tenderness and water‐soluble collagen content. The eating quality of beef obtained from different crossbreds was similar; however, the meat from PHF × LM and PHF × HER bulls had lower WBSF values than PHF × CH bulls. © 2016 Japanese Society of Animal Science     

Effect of apple cultivars and storage periods on the virulence of Neofabraea spp.

Bull’s eye rot is a typical quiescent postharvest apple disease in major fruit-growing areas. The susceptibility of different apple cultivars to Neofabraea spp. (N. vagabunda and N. malicorticis) was assessed, with Granny Smith showing the most resistance and Cripps Pink the most susceptibility. To assess the factors involved in conidial germination, Neofabraea spp. were grown on crude protein extracts (CPEs) collected from apple fruits at different storage periods. Fungal germ tube growth rate and pathogenic enzyme (cellulase and xylanase) activity were assessed. Results showed that CPEs collected after 2 and 4 months of storage progressively stimulated conidial germination and germ tube elongation, while a lesser effect was observed from CPEs after 1 month of storage. Xylanase proved to be the main degrading enzyme secreted by all the isolates, while cellulase was produced only by N. vagabunda isolates. Overall, the isolate ID02 was the most virulent, based on more rapid germ tube elongation and greater activity of the lytic enzymes.     

Effects of glutathione treatments during sperm washing and in vitro fertilization on the in vitro early development of embryos of Japanese Black cattle

This study was conducted to examine the effects of adding glutathione (1 mM) to media used for sperm washing and in vitro fertilization (IVF) on the improvement of early development of embryos produced using cryopreserved spermatozoa of the less IVF-competent bull (the one considered unqualified as spermatozoa supplier for the production of bovine blastocysts using IVF). The cryopreserved spermatozoa of this bull were characterized by normal motility and lower ATP content and blastocyst productivity than those of IVF-competent bulls. The addition of glutathione to the sperm washing medium was more effective in improving the productivity of blastocysts and ATP content than the addition of glutathione to the IVF medium or no glutathione addition at all (control). These results suggest that this simple method may be used to improve the potential of cryopreserved spermatozoa of less IVF-competent bulls to fertilize oocytes in vitro and to induce normal embryonic development after fertilization.     

精粗比条件下新疆芦苇在断奶初期奶公犊瘤胃内动态降解的研究

选用3头三月龄左右装有瘤胃瘘管的荷斯坦公犊,按3×3完全拉丁方试验设计,喂给三种日粮,日粮组成按精粗比分别表示为:Ⅰ:70∶30;Ⅱ:60∶40;Ⅲ:50∶50,研究不同日粮组成下新疆芦苇的DM、CP以及NDF在断奶初期奶公犊瘤胃内动态降解。结果表明:精粗比为60∶40日粮时,其CP的消失率和降解参数显著低于精粗比为70∶30和50∶50的日粮(P<0.05);其DM和NDF的消失率和降解参数显著高于精粗比为70∶30和50∶50的日粮(P<0.05)。     

牛蛙腐皮病病变皮肤亚显微结构的观察

为揭示牛蛙腐皮病的病理形态和发病的机理，采用透射电镜对病蛙不同病期的病变皮肤进行了观察，皮肤病变早期可见角化层脱落，状上皮细胞连接疏松，桥粒减少，随着病情发展细胞间隙增宽，桥粒进一步减少，细胞核固缩、溶解，细胞裂解；病变后期，表皮层崩解，真皮层胶原纤维溶解，炎症区有大量炎症细胞浸润，并可见吞噬了细菌的单核与中性粒细胞。     

Adfluvial and fluvial life history variations and migratory patterns of a relict charr, Salvelinus confluentus, stock in west-central Idaho, USA

Abstract –  Life history strategies and migratory patterns of 71 adult radio-tagged bull trout, Salvelinus confluentus , were studied in the Secesh River watershed within the South Fork Salmon River (SFSR) sub-basin in west-central Idaho, USA during 2003 and 2004. In both years, upstream migrations occurred during late June and early July, migrations into two spawning tributaries during late July and early August, spawning from mid-August through mid-September, and rapid downstream (postspawning) migrations from late August to mid-September. Primary over-wintering areas were Loon Lake, the lower Secesh River (downstream of Loon Creek), and the lower SFSR (downstream of the confluence with the Secesh River). Loon Lake evidently provides sufficient production to allow the adfluvial life history strategy to persist and predominate in the Secesh River, while the fluvial life history strategy was previously found to predominate in the nearby East Fork SFSR. Adfluvial, nonconsecutive-year migrations were the predominant life history strategy. Only seven fish made consecutive-year migrations to Lake Creek; however, only one of these fish, a female, utilised a spawning tributary in both years and showed spawning tributary fidelity. Three consecutive-year migrants and three in-season migrants showed over-wintering site fidelity by returning to Loon Lake in September, 2004. The life history variations observed for bull trout in the Secesh River watershed are similar to those observed for bull trout throughout their range and to those of other charr species worldwide, yet the development of distinct migration patterns demonstrate the adaptability of the species to a range of available habitats.     

肝素或钙离子载体诱导牛、羊冻精体外获能的研究

本文采用肝素或钙离子载体(I-A)诱导牛、羊冻精体外获能,根据对去透明带仓鼠卵的穿透情况评定获能效果,并用台盼蓝·姬姆萨(TG)染色观察了精子的死活及顶体状态.结果发现,牛、羊冻精经肝素或I-A处理后与卵子一起培养,2 h穿透仓鼠卵,4 h开始形成雄原核,6 h形成发育良好的雄原核.牛冻精用50μg/mL肝素及0.1μmol/L I-A处理效果最好,穿透率分别为72.8％和92.3％;羊则以20μg/mL肝素及0.3μmol/L I-A为宜,穿透率分别为73.1％和68.O％.咖啡因与I-A或肝素并用均有协同作用,能提高精子的穿透能力,并能促进卵内原核的发育.肝素和I-A均能在体外诱导牛、羊冻精的顶体反应,有顶体反应活精子百分率与穿透率呈强正相关,羊冻精比较脆弱,获能处理后活力较低.     

公牛精母细胞减数分裂与精子发生的研究

本研究用睾丸组织切片和细胞悬浮液制备的减数分裂与联会复合体标本，观察研究了公牛扁牛减数分裂及精子发生过程。与以往的研究结果不同的是，发现有些公牛扁牛的部分精母细胞能够完成减数分裂的全过程，直至出现精子。但在整个过程中伴随着大量异常，破坏了精子生成的机能。