Retracted: Study of testis histology and sperm morphology in the Bunnei,Barbus sharpeyi (Gunther, 1874) induced by luteinizing hormone‐releasing hormone analogue and carp pituitary extract

This study was conducted to determine the effects of hormone treatment on testis structure in Barbus sharpeyi, as well as the morphology of sperm examined by scanning electronic microscopy (SEM). Male B. sharpeyi were divided into three groups (three fish per group) and injected with luteinizing hormone – releasing hormone analogue (LHRH–A₂) or carp pituitary extract (CPE). The first and second groups were treated with 10 μg kg^?1 LHRH–A₂ and metoclopramide (MET), and their testis were sampled pre‐ and Poststripping respectively. The third group received 2 mg kg^?1 CPE and were killed pre‐stripping. Based on the histological results obtained, the testicular connective tissue of the lumen was thicker, and seminal vesicles were of a lower volume, in fish injected with CPE in comparison to the other groups. After treatment with LHRH–A₂ and MET, not all spermatozoa within the testis were ejaculated, and only a small amount of sperm was obtained by abdominal stripping. The highest and lowest diameters of connective tissue within lobules were observed in fish receiving CPE and LHRH–A₂ treatments respectively. There was a significant difference (P < 0.05) in lobule space between the fish injected with the CPE and the fish injected with the LHRH–A₂ and MET. The SEM results revealed that the spermatozoa of B. sharpeyi were composed of a spherical to elliptical head, a cylindrical midpiece, and a lengthy flagellum. In conclusion, it was found that injection with LHRH–A₂ and MET improved the spermatogenic process in comparison to injection with CPE.     

Fertilization rate,motility, lipid peroxidation and pH changes after chilled storage of rainbow trout (Oncorhynchus mykiss) eggs and spermatozoa by a RMPI medium

The aim of this study was to evaluate the effects of Roswell Park Memorial Institute (RPMI) 1,640 medium on chilled storage of eggs and spermatozoa of rainbow trout (Oncorhynchus mykiss). After 3 days of storage, eggs in RPMI 1,640 media (pH 8.2, 9 and 10), Cortland medium and coelomic fluid were inseminated with fresh spermatozoa (Experiment 1). Eggs in RPMI 1,640 medium at pH 8.2 shown the lowest thiobarbituric acid‐reactive substances (TBARS, 0.053 ± 0.003 nmol/ml) and pH changes (from 8.20 ± 0.01 to 8.18 ± 0.01), the highest fertilization rate (82 ± 3%). Undiluted and diluted spermatozoa at ratios of 1:2 and 1:9 with RPMI 1,640 media (pH 8.2, 9 and 10) and Cortland medium were inseminated with fresh eggs (Experiment 2). Spermatozoa in RPMI 1,640 medium at pH 9 (1:9) caused the lowest TBARS content (0.037 ± 0.002 nmol/ml) and pH changes (from 9.00 ± 0.01 to 8.98 ± 0.01), the highest fertilization rate (77 ± 2%) and motility parameters. Based on Experiments 1 and 2, eggs and spermatozoa were stored for another 3 days in RPMI 1,640 medium at pH 8.2 and 9 (1:9) respectively (Experiment 3). Fertilization rate of storage eggs and spermatozoa in Experiment 3 was 79 ± 5%, showing successfully storage of rainbow trout gametes with the same medium.     

Motility of sea urchin Paracentrotus lividus spermatozoa in the post‐activation phase

The characterization of sperm motility patterns, particularly post‐activation changes, is the first step in setting up species‐specific protocols involving gamete management and embryo production, for both aquaculture and laboratory research purposes. This study is aimed at the characterization of the sperm motility pattern of the purple sea urchin Paracentrotus lividus. Semen samples were individually diluted in artificial sea water for sperm motility activation. They were then incubated at 18°C for up to 24 hr. Motility was evaluated on dilution, and 1 hr, 3 hr and 24 hr after activation, by computerized analyser. The semen fertilization capacity was also evaluated. Under our experimental conditions (dilution 1:1,000 in artificial sea water plus 0.05% BSA, 18°C, in the dark), P. lividus semen remained viable for up to 24 hr, as the total motile sperm and the fertilization percentages did not change significantly during the incubation time. In contrast, the mean curvilinear velocity and the subpopulation of rapid sperm (those having a curvilinear velocity > 100 µm/s) slightly but significantly decreased after 3 hr, thereafter remaining unchanged for up to 24 hr after activation. In conclusion, our results show that diluted P. lividus semen can be used for a longer period than that of most fish species, with no need for motility inhibition procedures, supporting its wider use in laboratory research. In addition, the development of artificial fertilization protocols for aquaculture production is simplified by long‐lasting sperm motility.     

Sperm motility and seminal plasma characteristics in Barbus sharpeyi (Günther, 1874)

Spermatozoa concentration, ionic composition, osmolality, glucose and total protein contents of seminal plasma and sperm motility were determined in Barbus sharpeyi (Cyprinidae, Teleosotei). Spermatozoa concentration ranged from 9.77 to 20.20 × 10⁹ spermatozoa mL^?1. Osmolality (mOsmol kg^?1) and ionic contents (mM L^?1) of the seminal plasma were 274.5±9.0, 70.0±3.4 Na⁺, 28.8±0.9 K⁺, 101.7±3.1 Cl^?, 0.9±0.1 Mg²⁺ and 2.1±0.1 Ca²⁺ respectively. Total protein and glucose were 5.3±0.2 g L^?1 and 76.7±4.3 mM L^?1 respectively. Sperm motility was initiated in a hypo‐osmotic condition, composed of either an ionic (KCl or NaCl) or a non‐ionic (sucrose) activation medium. Duration of sperm motility was very short: <2 min after activation in distilled water. Percentage of motile spermatozoa was significantly higher in an activation medium containing NaCl compared with that of distilled water. An activating medium containing NaCl or KCl higher than 150 mM or sucrose higher than 275 mM totally inhibited the activation of sperm motility. Immediately after sperm activation, wave(s) propagated along the flagellum, but waves were restricted to the proximal part of the flagellum (close to the head) at 1 min post activation. Studied characteristics in the present study were compared with those of other cyprinids for understanding inter‐species differences.     

中华乌塘鳢精子的生物学特性及其超低温保存

对中华乌塘鳢精子激活的生理生态学进行了研究，探讨了中华乌塘鳢精子超低温保存方法，中华乌塘鳢精子激活的适宜盐度为５－１５；其精子的激活不仅与激活溶溶盐度有关，而且与激活溶液的离子成份有关，激活溶液中Ｋ＾＋的存在一定程度上对精子的活动有抑帛和；中华乌塘鳢精子对ＰＨ的适应性强，在ＰＨ５．５－９．５范围内，激活率均在７０９％以上，尤以中性及弱酸性条件下的激活率最高，在ＰＨ６．０时精子活力最好。使用筛选出的     

Single layer centrifugation of stallion spermatozoa consistently selects the most robust spermatozoa from the rest of the ejaculate in a large sample size

Reasons for performing study: An improvement in sperm quality after single layer centrifugation (SLC) has been seen in previous studies using small sample sizes (for example, n = 10 stallions). There is a need to investigate whether this improvement is repeatable over several breeding seasons with a larger number of stallions (n ≥ 30 stallions). Objective: To make a retrospective analysis of the results of SLC performed on more than 250 sperm samples (176 ejaculates) from 31 stallions in 3 consecutive breeding seasons. Methods: Sperm quality (motility, proportion of morphologically normal spermatozoa and the proportion of spermatozoa with undamaged chromatin) was assessed before and after SLC. Results: All parameters of sperm quality examined were significantly better in sperm samples after SLC than in their unselected counterparts (P<0.001 for each parameter). The yield of spermatozoa obtained after SLC was influenced by the type of extender used and also by the concentration of spermatozoa in the original ejaculate, with fewer spermatozoa being recovered when the loading dose contained a high concentration of spermatozoa. The optimal concentration was approximately 100 × 10⁶/ml. Sperm concentration in the samples loaded on to the colloid influenced the sperm yield while the type of semen extender affected sperm quality and survival. Furthermore, the scaled‐up SLC method was found to be suitable for use with a range of ejaculates, with similar sperm kinematics being observed for standard and scaled‐up preparations. Conclusions: SLC consistently improved the quality of stallion sperm samples from a large number of ejaculates. The method could be scaled‐up, allowing larger volumes of ejaculate to be processed easily from a wide range of stallions.     

七带石斑鱼精子的超微结构

利用透射电子显微镜技术研究了七带石斑鱼（Epinephelus septemfasciatus）精子的超微结构,结果发现：七带石斑鱼精子由头部和尾部（鞭毛）两部分组成,没有明显的中片,头部前段无顶体,主要有高电子密度的球形细胞核,核中主要是致密的染色质,还分布有一些核泡,核外由核膜包围,将核与细胞质分隔开,核膜最前端紧贴质膜,核后端有植入窝,近端中心粒和中心粒间体位于其中。头部后端为袖套结构,由质膜包围的袖套结构延伸到后部,使整个头部呈椭球形,质膜后端沿鞭毛形成一个袖套腔,袖套结构中分布有中心粒复合体、线粒体以及囊泡等。精子细胞的细胞膜不光滑,呈现小的皱折。七带石斑鱼精子的尾部细长,紧接于头部细胞核,穿过袖套腔,通过基体与核紧密结合,中心粒复合体也位于此,且近端中心粒和远端中心粒呈钝角,鞭毛的轴丝由此形成,基体与鞭毛间的过渡区轴丝无中央微管,呈“9+0”型。尾部的主要结构为轴丝,其轴丝为典型的“9+2”结构,其起始端开始于前端基体,轴丝外仅有少许细胞质。     

超低温冷冻保存对大黄鱼精子酶活性的影响

研究超低温(-196℃)冷冻保存对大黄鱼(Pseudosiaena crocea)精子内总ATP酶、肌酸激酶(CK)、琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽还原酶(GR)等酶活性的影响。运用试剂盒分别测定了冷冻前后大黄鱼精子内酶活性的变化。结果表明,经过超低温冷冻保存后,大黄鱼精子的活力下降,精子内GR活性从(4.42±0.29)U·L-1增加到(58.93±2.26)U·L-1(P<0.05);其它几种酶的活性均显著下降(P<0.05),总ATP酶、CK、SDH的活性分别从冻前的(60.16±5.88)U·mL-1、(11.91±0.76)U·mL-1和(51±2.16)U·mL-1下降到(3.54±0.37)U·mL-1、(10.22±0.32)U·mL-1和(31.5±2.08)U·mL-1;LDH、SOD和CAT活性从冻前的(7 806.44±110.11)U·L-1、(42.65±1.56)U·mL-1和(119.91±8.10)U·mL-1下降到(2 654.13±70.06)U·L-1、(31.99±1.57)U·mL-1和(55.87±2.32)U·mL-1。超低温冷冻保存对大黄鱼精子活力和精子酶活性均有显著影响。     

The Comet assay for detection of DNA damage in canine sperm

Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno‐, terato‐ or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A—median: 31.55%, interquartile range (IQR): 30.18–38.01; group B—median: 0.90%, IQR: 0.65–1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = ?.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non‐normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs.     

超低温冷冻对俄罗斯鲟精浆和精子酶活性的影响

研究了超低温冷冻保存(-196℃)对俄罗斯鲟(Acipenser gueldenstaedti)精浆和精子中总三磷酸腺苷酶(AT-Pase)、琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)、肌酸激酶(CK)等酶活性的影响,并分别测定了冷冻前后俄罗斯鲟精浆和精子中酶的活性。结果显示,经过超低温冷冻保存后,俄罗斯鲟精子活力下降,精子内各酶活性均显著降低,添加冷冻保护液组精子中总ATPase、SDH、LDH和CK的活性分别从(198.47±14.43)U/mL、(30.00±2.65)U/mL、(6 982.29±24.32)U/L和(1.94±0.05)U/mL下降至(110.19±2.32)U/mL、(16.33±2.08)U/mL、(5 122.93±195.07)U/L和(1.49±0.14)U/mL。未添加抗冻剂组则分别下降至(2.25±0.33)U/mL、(11.67±0.58)U/mL、(4 488.04±78.33)U/L和(1.16±0.02)U/mL;精浆中酶的活性均显著升高,添加冷冻保护液组精浆总ATPase、SDH、LDH和CK活性分别从(12.70±0.57)U/mL、(7.50±0.71)U/mL、(2017.26±116.81)U/L和(2.93±0.59)U/mL升高至(92.49±5.18)U/mL、(13.33±0.58)U/mL、(3 688.97±172.67)U/L和(4.39±0.24)U/mL,未添加抗冻剂组则分别上升至(200.27±12.97)U/mL、(24.67±3.06)U/mL、(6 124.40±329.14)U/L和(5.20±0.16)U/mL。结果表明,超低温冷冻对俄罗斯鲟精浆和精子中酶活性及精子活力均有较大影响。