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.     

紫茎泽兰对萨能奶山羊血常规及重要脏器病理学的影响

紫茎泽兰(Eupatorium adenophorum)在中国作为入侵物种,可造成食草动物中毒死亡,并给畜牧业带巨大的损失。为研究反刍动物采食紫茎泽兰后的中毒机理,本研究探讨了日粮中添加不同剂量的紫茎泽兰对萨能奶山羊(Saanen goat)血常规及肝、脾、肾的病理学影响。选择16只5~6月龄健康萨能奶山羊,随机分成对照组(日粮中未添加紫茎泽兰)和试验Ⅰ、Ⅱ、Ⅲ组(日粮配比中分别添加紫茎泽兰40%、60%和80%),每组4只。试验Ⅰ、Ⅱ、Ⅲ组分别饲喂含40%、60%和80%紫茎泽兰的混合日粮3个月,每两周检测一次血常规,试验末屠宰奶山羊,并观察其肝、肾、脾的病理学变化。结果表明,与对照组相比,各试验组血液中的白细胞(white blood cell,)和中性粒细胞(neutrophil,NEUT)均显著升高(P0.05),试验后期极显著高于对照组(P0.01);而各试验组血液中淋巴细胞(lymphocyte,LY)与血红蛋白(hemoglobin,HGB)均呈下降趋势,试验后期均极显著低于对照组(P0.01)。肝和脾肿大,肝组织出现淤血,肝细胞发生水泡变性及脂肪变性等,肾组织伴有出血、坏死、颗粒变性及水泡样变性的现象。因此,本研究结果说明紫茎泽兰对萨能奶山羊血液指标有显著影响,且长期摄入紫茎泽兰可引起动物肝、肾、脾等主要实质器官不同程度的损伤。     

检疫犬基地建设与发展策略探讨

检疫犬应用技术作为一种特殊的检验检疫手段,目前已被30多个国家或地区应用于检验检疫领域。许多国家或地区已建有自己的国家级工作犬训练基地。近年来,由于出入境业务量的迅速增长,我国对检疫犬数量的需求逐年攀升,明确要建成2个国家级检疫犬繁殖训养中心。检疫犬基地作为培养检疫犬的重要场所,其基地面积、位置规划、人力资源、种犬储备、幼犬来源等的科学合理规划,对保证其可持续发展具有重要意义。检疫犬基地的可持续发展,需要准确定位功能需求,搞好种犬繁育,统筹储备人力资源,建设生态环保基地,挖掘和利用社会资源,以及拓展其它功能建设,从而做到行业引领。     

2018年安徽省部分地区A型禽流感流行率横断面研究

为了解安徽省A型禽流感病毒感染情况,2018年4—5月,在安徽省北部、中部和南部7个市,随机选取各类型场点173个,在每个场点内随机采集35份咽喉/泄殖腔和环境拭子样品,使用荧光RT-PCR进行A型、H5亚型、H7亚型流感病毒核酸检测。结果显示：在4个养殖场、3个活禽批发市场和17个农贸市场检出了A型禽流感病毒,场点阳性率分别为2.9%（4/140）、75.0%（3/4）、80.9%（17/21）,未检出H5和H7亚型流感病毒核酸;A型禽流感群体流行率为11.2%（95%CI：6.5%~15.8%）,个体流行率为1.2%（95%CI：0.9%~1.5%）;活禽交易市场内,禽群感染A型禽流感病毒的风险较高（OR=136,95%CI：32.7~549.4）。结果表明,A型禽流感病毒在养殖和交易环节存在一定的污染,而在活禽交易市场的污染更严重,是今后禽流感防控的重点,需继续加强活禽交易市场的管理和消毒灭源工作。     

犊牛魏氏梭菌的分离鉴定及致病性研究

2017年1月,山东省某规模化奶牛场10～20日龄犊牛出现腹泻、拉血便、呼吸困难等症状,部分犊牛突然死亡。通过流行病学调查和尸体剖检,初步怀疑为魏氏梭菌感染。采集病料进行病理组织学检查、细菌分离鉴定、生化试验、16S及毒素基因检测等,证明疫情由A型魏氏梭菌所致。对死亡犊牛的肠内容物及分离菌株进行致病性研究,发现肠内容物及分离菌株具有较强的致病性。药敏试验结果显示,该菌对青霉素、头孢噻呋、恩诺沙星等表现极敏感。根据病原确诊情况及药敏试验结果,指导临床用药,及时控制了牛场疫情。     

干旱区工业废渣粉煤灰回填材料拌合物的流动性能研究

针对干旱区存在大量的粉煤类废弃物及回填工程中作业面受限或结构物死角存在压实不到位的工程问题,提出以粉煤灰为主要成分、以及部分砂、水泥的流动化回填材料,分析拌合物流动性能受材料掺量、时间影响的变化规律,建议采用坍落扩展度作为流动性能评价指标,水泥掺量对流动性能影响甚微,坍落扩展度与水固比成线性关系;同一种集料,不同粉煤灰掺量拌合物达到相同的流动性能,水固比呈先降后升的变化趋势;拌合物坍落扩展度在前60min内降低显著,随时间变化,坍落扩展度趋于一致,拌合物中集料颗粒越小,保持流动性的能力越强.     

鳢肠对棉铃虫生长发育的影响

通过饲料混毒法测定鳢肠对棉铃虫生长发育的影响。结果表明,随饲料中鳢肠比例的加大和棉铃虫幼虫取食时间的延长,鳢肠对棉铃虫幼虫的生长发育具有明显的抑制作用。幼虫取食鳢肠比例高的饲料后,棉铃虫蛹质量显著降低,但羽化率和单雌日均产卵量与对照处理差异不显著;给棉铃虫幼虫饲喂鳢肠明显延长了幼虫和成虫历期,但对蛹历期的影响较小。     

Identification of organic acid-related genes and their expression profiles in two pear (Pyrus pyrifolia) cultivars with difference in predominant acid type at fruit ripening stage

‘Yandangxueli’ is a pear cultivar with predominant citric acid in the ripe fruit, different from most of pear cultivars such as ‘Gengtouqing’ in which malic acid is the predominant acid type. It was found that ‘Yandangxueli’ accumulated citric acid for three times against that in ‘Gengtouqing’ at fruit ripening stage. To investigate the mechanism of citric acid accumulation in ‘Yandangxueli’, organic acids content, gene expression and enzyme activity were studied in both cultivars. Five genes, Pp:mtCs, Pp:cyAco, Pp:cyIdh, Pp:mtMdh and Pp:cyMe which encoded citric synthase (CS), cytosolic aconitase (cyACO), NADP-dependent isocitrate dehydrogenase (NADP-IDH), NAD-dependent malate dehydrogenase (NAD-MDH) and NADP-dependent malic enzyme (NADP-ME) respectively, were identified from pear fruit. Their expression profiles and the corresponding enzyme activities were determined throughout fruit development in both cultivars. Results from these enzymes indicated that there were no strict relationship between gene expression, enzyme activity and citric acid accumulation. Expression analysis for two Py:vVAtp genes encoding vacuolar H⁺-ATPase A subunit and one Py:vVpp gene encoding Vacuolar H⁺-pyrophosphatase showed that they were all with up-regulated expression at the later development stage of ‘Yandangxueli’ but with down-regulated expression in ‘Gengtouqing’. Therefore, it is concluded that the different ability in citric acid transportation and storage might be involved in the high citric acid content in ‘Yandangxueli’.     

Dvl1及其突变体腺病毒载体的构建与在MSCs中表达鉴定

Dvl1及其结构缺失突变体ΔDIX（dsh and axin）和ΔDEP（dsh,Egl-10 and pleckstrin）腺病毒载体的构建并验证其在MSCs（mesenchymal stem cells）中的表达情况。将目的基因定向克隆至pAdTrack-CMV穿梭载体上,并经PmeⅠ线性化后在BJ5183细菌中与pAdEasy-1骨架质粒同源重组,获得重组腺病毒载体,在QBI-293A细胞中包装及扩增,实时荧光定量PCR和Western bolt验证Dvl1在MSCs中的表达情况。经PCR、PacⅠ单酶切鉴定及测序分析,成功构建Ad-Dvl1、Ad-ΔDIX和Ad-ΔDEP腺病毒载体,目的基因序列与GenBank报道一致,并选出150 MOI为最适感染MSCs的感染复数,成功构建了Ad-Dvl1、Ad-ΔDIX和Ad-ΔDEP腺病毒载体,并获得高滴度的病毒子,qPCR和Western blot证实Dvl1在MSCs中高表达并增加了β-catenin在细胞中的累积,为进一步研究Dvl1在MSCs迁移过程的作用奠定了基础。