某牛场犊牛大肠杆菌病的实验室诊断与药敏试验

2018年3月份,黑龙江省齐齐哈尔市某规模化奶牛场发生犊牛腹泻,为了确诊其病因,无菌采取10头腹泻犊牛粪便进行细菌分离鉴定、生化试验、PCR鉴定和药敏试验等。生化结果表明:符合大肠杆菌生化特性;PCR结果表明:引物能有效扩增大肠杆菌OmpA、K99、Sta、irp2、FyuA基因;药敏结果表明:大肠杆菌对恩诺沙星、左氟沙星、头孢曲松高度敏感;10头犊牛均为大肠杆菌感染,本研究为该牛场犊牛腹泻的病因分析和防治提供了指导。     

母鼠孕期和哺乳期连续染毒双酚A对子代雄鼠睾丸发育的影响

旨在探究双酚A (BPA)对子代雄鼠睾丸发育的影响。本研究将8周龄体重18～22 g的SPF级母鼠随机分为7组,每组4个重复,每个重复5只,A组每天给予普通蒸馏水,B组每天饮水给予0.05 mg·kg^-1 BPA,C组每天饮水给予0.5 mg·kg^-1 BPA组,D组每天饮水给予5 mg·kg^-1 BPA,E组每天饮水给予10 mg·kg^-1 BPA,F组每天饮水给予20 mg·kg^-1 BPA,G组每天饮水给予50 mg·kg^-1 BPA。F0代母鼠自怀孕起直至F1代子鼠断奶止饮水染毒BPA,F1代雄鼠于断奶(21日龄)处死。ELISA结果显示,母鼠染毒BPA剂量5 mg·kg^-1以上时,子代血清和睾丸组织BPA含量显著增加(P<0.05)。睾丸器官指数测定结果证明,染毒BPA剂量大于等于20 mg·kg^-1可导致子代雄鼠睾丸指数显著增大(P<0.05)。H&E染色显示,母鼠染毒BPA剂量大于等于10 mg·kg^-1可导致睾丸生精小管萎缩,小管间隙变大。彗星试验结果证明,染毒BPA大于等于5 mg·kg^-1可使子代睾丸细胞核DNA损伤显著上升(P<0.05)。免疫组化结果显示,母鼠染毒BPA剂量大于等于20 mg·kg^-1时子代睾丸雄激素受体(AR)表达量显著减少(P<0.05)。转录组测序结果显示,母鼠染毒50 mg·kg^-1BPA后子代雄鼠睾丸剪切体代谢通路U5 snRNA亚基编码基因Snrnp40上调,剪切体通用蛋白组件编码基因Hnrnpu下调,导致mRNA转录后修饰第一步受阻,荧光定量PCR结果与转录组测序结果一致,证实了转录组结果。结果表明,母鼠暴露于低剂量BPA可以引起子代睾丸发育异常,其分子机制可能与剪切体进行mRNA转录后修饰第一步反应受阻有关。     

Transient glucose malabsorption in two horses fact or artefact?

Two horses, presented for investigation of chronic weight loss despite normal to increased feed intake, had flat oral glucose absorption curves, suggesting malabsorption. The cause of the apparent malabsorption was not evident grossly or on light microscopic examination of the intestinal tract. Both horses survived long term and at follow-up examination had regained weight and their capacity to absorb glucose. These cases illustrate that flat glucose absorption curves may occur in horses with no obvious intestinal lesions, that they may revert to normal and that the results of these tests should be interpreted with caution.     

乳粉中发酵乳杆菌CECT5716微生物检测方法的建立

近年来乳粉中添加益生菌逐渐成为研究热点之一,为保证产品质量和安全,建立乳粉中发酵乳杆菌CECT5716的微生物检测方法。结果表明：在以缓冲蛋白胨水为稀释液进行10 倍系列稀释后,采用MRS平板涂布法,（36±1） ℃厌氧培养（48±2） h后,乳粉中发酵乳杆菌可准确计数,且能与乳粉中添加的双歧杆菌属细菌明显区分;该方法较GB 4789.35—2016《食品安全国家标准 食品微生物学检验 乳酸菌检验》方法更适合发酵乳杆菌计数,具有区分度高、培养时间短、操作简单、重复性好、准确性高的优点;该方法增加了菌株16S rDNA序列测定,以进一步对发酵乳杆菌进行分子鉴定;通过对20 份自制样品进行检测评价,表明所建立的方法与国标方法的准确性无显著性差异（P=0.127）。     

中国美利奴羊(新疆型)无角类型培育理论依据及测交试验结果

绵羊角的有无是由或主要由一个基因位点（Locus）上的三个等位基因（H、h’、h）所决定。三个等位基因的遗传方式是：H为显性无角基因，对h为完全显性，即Hh基因型均表现无角性状；h为隐性有角基因，即hh基因型均表现有角性状；h，为一个频率很低的基因，对h为不完全显性，即h＇h基因型仍表现为有角性状；但H对h’基因为不完全显性，即Hh’基因型可能出现角痕（角突或角基）。而h’h’基因型公羊表现为小角、发育残缺不全的矫型角，母羊表现为无角。培育无角革的关键是选择使用无角公羊，通过测交试验能确定无角公羊基因的纯合度，从而决定如何使用。     

暴发高致病禽流感疫区鸟类禽流感的血清学调查

本研究应用血凝抑制试验(H I),2004年初H 5亚型禽流感在中国内地发生前后,于湖北、广西二疫区市县的生态系中,自健康的鹰、鸽子、野鸭和鹌鹑中采集了54份血清,对其进行了血清学检测,并将获得的血清学检测数据进行了统计学分析。分析结果显示,在总共采集的54份血清样品中,抗H 6亚型禽流感病毒(H 6)抗体、抗H 9亚型禽流感病毒(H 9)抗体、抗H 5亚型禽流感病毒(H 5)抗体阳性率分别为64.8%、75.9%、29.6%。推断,这一时期,H 6亚型禽流感病毒(H 6)、H 9亚型禽流感病毒(H 9)和H 5亚型禽流感病毒(H 5)共循环于该生态系中,其中H 9、H 6亚型禽流感病毒长期、稳定循环于该生态系中,为该生态系禽流感主要流行血清型。血凝抑制试验检测结果发现,H 5亚型禽流感病毒抗体滴度较低,最高仅为640×,其阳性率为29.6%,推断其可能为新近传入的病毒或新近重组变异出现的新病毒,可能为与该次禽流感暴发相关的新病毒,这一推断还有待于通过对病毒基因序列的分析而做进一步验证。     

Thyroid Hormone-disrupting Effects and the Amphibian Metamorphosis Assay

There are continued concerns about endocrine-disrupting chemical effects, and appropriate vertebrate models for assessment of risk are a high priority. Frog tadpoles are very sensitive to environmental substances because of their habitat and the complex processes of metamorphosis regulated by the endocrine system, mainly thyroid hormones. During metamorphosis, marked alteration in hormonal factors occurs, as well as dramatic structural and functional changes in larval tissues. There are a variety of mechanisms determining thyroid hormone balance or disruption directly or indirectly. Direct-acting agents can cause changes in thyroxine synthesis and/or secretion in thyroid through effects on peroxidases, thyroidal iodide uptake, deiodinase, and proteolysis. At the same time, indirect action may result from biochemical processes such as sulfation, deiodination and glucuronidation. Because their potential to disrupt thyroid hormones has been identified as an important consideration for the regulation of chemicals, the OECD and the EPA have each established guidelines that make use of larval African clawed frogs (Xenopus laevis) and frog metamorphosis for screening and testing of potential endocrine disrupters. The guidelines are based on evaluation of alteration in the hypothalamic-pituitary-thyroid axis. One of the primary endpoints is thyroid gland histopathology. Others are mortality, developmental stage, hind limb length, snout-vent length and wet body weight. Regarding histopathological features, the guidelines include core criteria and additional qualitative parameters along with grading. Taking into account the difficulties in evaluating amphibian thyroid glands, which change continuously throughout metamorphosis, histopathological examination has been shown to be a very sensitive approach.     

转Xa23基因水稻的白叶枯病抗性及其遗传分析

在分离克隆抗白叶枯病基因Xa23研究中获得大量转基因水稻材料。为了系统研究转Xa23基因水稻的抗病稳定性和遗传模式, 本文通过逐株进行抗白叶枯病接种鉴定、PCR和Southern blot分子检测, 对一批转Xa23基因水稻植株进行了T₀代到T₂代的跟踪分析。结果表明, Xa23基因的整合和表达, 使感病受体品种牡丹江8号获得抗病性。由于Xa23基因插入受体基因组的位点不同, 同是单拷贝插入的转基因T₀代抗病植株, 其抗病程度有明显差异。T₀代植株的抗病程度, 可以准确、稳定地遗传到T₁代和T₂代。单拷贝转基因植株分离群体的抗感植株分离比接近3∶1, 表明转Xa23基因遵循孟德尔单基因遗传模式。已获得2个纯合的单拷贝转基因抗病株系, 它们的抗病程度稍有差别, 将用于外源基因插入位置效应分析和杂交稻抗病育种。     

基于AHP-CRITIC法的正交设计优选贞芪颗粒提取工艺及促生长作用初步研究

（目的）开发一种具有促生长作用的新中兽药贞芪颗粒,优化其提取工艺并对其促生猪生长作用进行初步研究,为其工业化生产提供参考。（方法）以总多糖、黄芪甲苷、毛蕊异黄酮葡萄糖苷、特女贞苷、红景天苷、干膏得率作为评价指标,采用层次分析法（AHP）、基于指标相关性的权重确定方法（CRITIC）、AHP-CRITIC混合加权法确定各评价指标的权重系数,根据其正交试验结果优化贞芪颗粒提取工艺参数。将100头保育猪随机分为5组,通过在日粮中添加不同剂量的颗粒,连续喂养30天,与空白组和阳性对照组对比,计算平均日增重（ADG）、平均日采食量（ADFI）和料肉比（F: G）。（结果）AHP-CRITIC混合加权法科学优选出最佳提取工艺参数为复方药材加12-13倍质量的水提取3次（第一次13倍）,每次2.5 h。3批验证试验综合评分均值为98.91,RSD为0.79%。经过30天饲喂,高、中、低剂量组和阳性对照组的料重比无显著性差异（P>0.05）,但都显著低于空白对照组（P<0.05）,其中中剂量组的料重比最低。（结论）AHP-CRITIC混合加权法确定的权重系数客观、真实,优选出的贞芪颗粒提取工艺经验证,合理、稳定,重复性好,此工艺条件下制备的颗粒对生猪具有促进生长的作用。     

Influence of acute exposure to a low dose of systemic insecticide fipronil on locomotor activity and emotional behavior in adult male mice

Fipronil (FPN) is a systemic insecticide that antagonizes the gamma-aminobutyric acid type A (GABA_A) receptors in insects. Recently, adverse effects of FPN on mammals have been reported, but most of those were caused by high doses of FPN and additives in the products. We investigated the effects of low-dose pure FPN on the emotional behavior of mice. Nine-week-old male mice conducted behavioral tests 24 hr after FPN administration by gavage at doses of 0.05 or 5 mg/kg based on the no-observed-effect level (NOEL), showed a significant increase in locomotor activity and dose-dependent responses on the time they spent in the central zone in the open field test. Pure FPN below the NOEL dose may affect the emotional behavior of mice.