Effects of Addition of Tissue‐Type Plasminogen Activator in In Vitro Fertilization Medium on Bovine Embryo Development and Quality

Plasminogen activators/Plasmin system plays pivotal role in regulating reproductive functions of mammals. Here, we examined the effects of modification of in vitro fertilization medium (IVF medium) with the addition of tissue‐type plasminogen activator (t‐PA), on bovine embryo development and quality, assessed by quantification of expression of various genes related to metabolism, oxidation, implantation and apoptosis. In addition, plasminogen activator activity (PAA) and plasminogen activator inhibition (PAI) were measured in the spent media. After conventional IVM, 2016 cumulus‐oocyte complexes (COCs) were divided into four groups with modified composition of the IVF medium containing t‐PA and/or its inhibitor epsilon‐aminocaproic acid (control, t‐PA, t‐PA+ε‐ACA, ε‐ACA). Presumptive zygotes were cultured for 8 days in synthetic oviductal fluid (SOF) medium; gene expression studies were carried out on morulae and blastocysts. t‐PA alone significantly suppressed cleavage and blastocyst formation rates, but this effect was neutralized by the addition of ε‐ACA. PAA in the treated group was significantly reduced by ε‐ACA, but without total elimination. Significant differences were detected in the expression of genes related to apoptosis and/or cell cycle arrest (BAX, BCL2L1, KAT2B) between embryos produced in t‐PA‐modified media and controls, giving an overall notion that the inferior developmental competence of treated embryos may be attributed to apoptotic phenomena induced by t‐PA. In conclusion, it appears that excessive t‐PA content in the IVF media, suppresses blastocyst formation rate, possibly due to induction of apoptotic phenomena.     

Impact of 24-h Cooling Prior to Freezing on the Survival of Domestic Cat (Felis catus) Epididymal Sperm

The aim of this study was to investigate the impact of a 24-h cooling period prior to freezing on domestic cat epididymal sperm viability. Fifteen tomcats were submitted to routine orchiectomy and sperm samples were retrieved from both epididymides in a Tris–glucose–20% egg yolk extender. For each tomcat, the diluted sperm was split into two equal volumes and cooled to 5°C at a rate of 0.5°C/min; one sample for 60 min (control) and the other for 24 h (cooled). After the cooling period, samples from both groups were frozen using an identical freezing protocol. Sperm samples were evaluated in three different periods: immediately after harvesting, after cooling at 5°C for 24 h (cooled group) and after freezing–thawing of control and cooled groups. Evaluations consisted of sperm motility and progressive status, sperm morphology and plasma membrane integrity (PMI) using two fluorescent probes. After cooling for 24 h, a decrease (p < 0.05) in sperm motility, progressive status and PMI was observed when compared to sperm samples immediately after collection. Comparing the results obtained after thawing, no difference (p < 0.05) was found regarding sperm motility, progressive status, PMI and sperm morphology between control and cooled groups. The results from the present study show that cooling cat epididymal spermatozoa at 5°C for 24 h prior to freezing does not lead to major damage of spermatozoa impairing the freeze–thaw process.     

Are ovine fenugreek (Trigonella foenum‐graecum) staggers and kangaroo gait of lactating ewes two clinically and pathologically similar nervous disorders?

Fenugreek staggers has occurred in sheep in Victoria, as both an acute and a chronic syndrome. Signs included quadraparesis, a high stepping fore limb gait and a 'bunny-hopping' hind limb gait. Changes consistent with acute oedema were found in the brain and spinal cord of acute cases, and Wallerian degeneration in the peripheral nerves of chronic cases. Kangaroo gait occurred in ewes in New South Wales, and the clinical signs and microscopic changes were remarkably similar to those of fenugreek staggers. Although the diet associated with each is different the causal agent may be the same.     

日光温室内反光膜补光的实践与理论探析

试验结果表明 ,温室后墙悬挂反光膜进行补光后 ,对温室内测定点处最大光照度值无影响 ,但平均提高测定点处水平光照度值 15 4 8% ;温室内凌晨时气温、地温、墙温和被反光膜遮掩处墙温分别比对照温室低 1 1℃、1 5℃、0 9℃和 1 8℃ ,但正午时分别比对照温室高 1 6℃、1 4℃、0 3℃和低 11 4℃。温室经济产量增加 7 4 % ,当期经济效益增加 2 1元 /m2 。     

Deleterious Effects of Progestagen Treatment in VEGF Expression in Corpora Lutea of Pregnant Ewes

The aim of the current study was to determine the possible effects of progestagen oestrous synchronization on vascular endothelial growth factor (VEGF) expression during sheep luteogenesis and the peri‐implantation period and the relationship with luteal function. At days 9, 11, 13, 15, 17 and 21 of pregnancy, the ovaries from 30 progestagen treated and 30 ewes cycling after cloprostenol injection were evaluated by ultrasonography and, thereafter, collected and processed for immunohistochemical evaluation of VEGF; blood samples were drawn for evaluating plasma progesterone. The progestagen‐treated group showed smaller corpora lutea than cloprostenol‐treated and lower progesterone secretion. The expression of VEGF in the luteal cells increased with time in the cloprostenol group, but not in the progestagen‐treated group, which even showed a decrease between days 11 and 13. In progestagen‐treated sheep, VEGF expression in granulosa‐derived parenchymal lobule capillaries was correlated with the size of the luteal tissue, larger corpora lutea had higher expression, and tended to have a higher progesterone secretion. In conclusion, the current study indicates the existence of deleterious effects from exogenous progestagen treatments on progesterone secretion from induced corpora lutea, which correlate with alterations in the expression of VEGF in the luteal tissue and, this, presumably in the processes of neoangiogenesis and luteogenesis.     

Wheat-sensitive enteropathy in Irish setter dogs: possible age-related brush border abnormalities

The pathogenesis of a wheat-sensitive enteropathy was explored in a litter bred from two Irish setters with a naturally occurring enteropathy. Jejunal biopsies from all eight progeny exhibited morphological changes comparable to those in the parents, while biochemical abnormalities appeared to be related to age. In biopsies obtained from the first group of four dogs at eight months, the activities of alkaline phosphatase and of leucyl-2-naphthylamidase were almost undetectable while disaccharidases were unaltered. In contrast, analytical subcellular fractionation of biopsies obtained from the second group of four dogs at nine months showed that specific activities now reflected a major deficiency of brush border alkaline phosphatase, and normal brush border leucyl-2-naphthylamidase accompanied by elevated soluble activity. Further studies are indicated to determine whether these findings represent an age-related abnormality affecting specific microvillus membrane proteins.     

小鼠血清抗磺胺二甲氧嘧啶抗体的研制

将磺胺二甲氧嘧啶(SDM)与载体牛甲状腺球蛋白(BTG)偶联,构建SDM完全抗原SDM-BTG。用SDM-BTG免疫小鼠,诱导小鼠产生血清抗SDM抗体。ELISA检测表明:SDM-BTG免疫小鼠所产生的多克隆抗体与SDM-BSA包被抗原的结合有较强的特异性和较高的亲和力。