Pathological analysis of batch safety testing of veterinary vaccines using small laboratory animals

Batch safety tests (BSTs) of veterinary vaccines are conducted using small laboratory animals to assure the safety of vaccines according to several criteria, including clinical signs and change in body weight. Although the latter is used as an evaluation index in BSTs, there have been no reports on the internal changes that affect body weight during the test period. Therefore, we analyzed BST via pathological examination of the tested animals. Here, BSTs were performed for 176 batches using mice and 126 batches using of guinea pigs. Most of the gross findings could be classified into four lesion types (nodules, adhesions, ascites, no apparent signs), with only one vaccine inducing lesions that could not be classified into any of these four types. Histopathological examination revealed that the reactions caused by BST were pyogenic and/or granulomatous inflammation. Nodular or adhesive lesions comprised more severe pyogenic granulomatous inflammation than ascites or cases with no apparent gross lesions. These nodular or adhesive lesions were more frequently induced by vaccines that contained an adjuvant than by vaccines that did not contain an adjuvant. The cases with “exceptional” gross findings histologically presented severe necrosis of the hematopoietic system. Additional testing showed that these “exceptional” lesions were induced when a specific type of light liquid paraffin was injected along with other vaccine additives. Our results show that body weight loss and/or lesions during BST were induced by proinflammatory properties of the tested vaccines and that BST is a sensitive method for detecting unexpected effects of vaccine components.     

Carrier rate of the c.235G>C mutation in the bovine isoleucyl-tRNA synthetase (IARS) gene of Japanese Black cows at Kagoshima prefecture, Japan,and analysis of the metabolic profiling and reproductive performance of heterozygous cows

Bovine isoleucyl-tRNA synthetase (IARS) disorder, a major cause of weak calf syndrome, is caused by a homozygous missense (c.235G>C) mutation in the bovine IARS gene of Japanese Black (JB) cattle, which was identified in 2013. However, the extent to which the carrier rate has changed at Kagoshima prefecture, Japan, and whether the carrier status is associated with any clinical or reproductive problems, have yet to be ascertained. In this study, using a real-time polymerase chain reaction-based genotyping assay, we determined the carrier rate in a regional JB cow population at Kagoshima prefecture. Comparative analyses were performed on the metabolic profile test (MPT) results and reproductive performance data obtained for heterozygous carrier and homozygous wild-type cows. In 2009 and 2018, DNA samples were collected from 130 and 462 clinically healthy JB cows, respectively, in Kagoshima prefecture. MPT results and reproductive performance data were evaluated for 62 cows, comprising four heterozygous carriers and 58 wild-type cows. Genotyping revealed that the carrier rate was 6.9% in 2009 and 1.5% in 2018, the difference of which was statistically significant (P<0.005). There were no statistically significant differences between the carrier and wild-type cows with respect to either MPT results or reproductive performance, indicating that the carrier cows have necessary IARS activity to maintain minimal health and reproductive potential.     

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.     

不同氨基酸对泰万菌素发酵影响的研究

通过单因素试验考察了泰万菌素发酵培养基中不同氨基酸种类对其发酵过程的影响,并利用正交试验对发酵培养基中3种主要氨基酸的用量配比进行了优化,结果表明发酵培养基加入0.2%亮氨酸、0.1%色氨酸、0.1%甘氨酸对泰万菌素发酵转化率影响显著,发酵摇瓶效价从14000μ/mL提高到18000μ/mL,效价提高了28.5%,完成了对泰万菌素提高发酵转化率和发酵单位的目的,并为大规模商业化生产泰万菌素奠定了良好的基础。     

我国和欧盟兽药质量监督抽检工作机制对比研究

为更好地完善我国兽药质量监督抽检计划,通过对欧盟药品抽检管理工作机制的研究,本文分析了欧盟和我国兽药监督抽检管理方式的不同,从抽检责任部门、抽检目的、抽检计划内容、抽检计划制定流程、抽检品种的遴选和检测参数的确定等6方面进行了对比分析,探讨我国兽药质量监督抽检工作的管理中可以借鉴的方面。     

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。     

鹅大肠埃希菌的分离鉴定及系统进化树分析

为对发病鹅感染致病性大肠埃希菌的临床治疗提供指导并防控致病性大肠埃希菌感染,本研究无菌采集腹泻鹅及病死鹅脾脏、肝脏等组织器官进行病原菌分离培养、染色观察、生化鉴定、致病性试验,对菌株部分基因进行测序、构建遗传进化树并进行药敏试验。结果显示,普通琼脂培养基上生长出灰白色、圆形、凸起、边缘整齐菌落,麦康凯琼脂培养基上长出玫红色、光滑圆润菌落;分离菌株经革兰氏染色,镜下可见革兰氏阴性菌,两端钝圆、大小中等,多呈单个存在的杆状菌;致病性试验表明,该分离菌株对小鼠、雏鹅均有较高致死率;遗传进化树结果显示,分离菌株与患者粪便分离株（GenBank登录号：CP024992.1）同源性最高,达99.5%;体外抑菌试验发现菌株耐药情况较严重,对大多数抗菌药均有较强抗性,仅头孢噻呋对该分离株有较强的抑菌作用,合理用药后病情得到有效控制。本研究为有效防治鹅大肠埃希菌病提供了理论依据,对规模化鹅场防控大肠埃希菌等其他细菌性疾病具有重要价值。     

低血糖生成指数液态奶的开发及测试

为获得营养均衡的调制牛奶,满足人体的多种营养需求,添加多种维生素和矿物质,设计出一款适用于糖尿病人、亚健康人群、体重管理、皮肤管理等人群饮用的低血糖生成指数（Glycemic Index,GI）的牛奶。稳定性试验结果表明,静置12 天后,牛奶无显著脂肪上浮,体系稳定性良好;人体GI值试验结果表明,调制牛奶的餐后血糖值上升和下降均较葡萄糖餐后血糖值平稳,有较好稳定餐后血糖的效果,表明开发的调制牛奶为低GI食品（GI值=22.86）。人体饱腹感试验结果显示,和普通纯牛奶对比,调制牛奶具有显著提高饱腹感的效果。     

四川凉山地区奶牛乳房炎主要病原菌分离鉴定及药敏情况分析

通过开展凉山地区奶牛乳房炎主要病原菌的分离鉴定和药敏情况试验,为指导凉山地区奶牛场临床合理使用抗生素以及提高抗生素的防治效果提供理论指导。通过采样、增菌培养、细菌鉴定、药敏试验、数据统计分析等方法,结果发现193 份患有乳房炎的乳样中有134 份分离出12 种病原菌,其中65 份检出两种及以上菌,占检出数的48.50%﹔共分离出234 株不重复菌株。分离率居前五位为：葡萄球菌63 株（26.9%）、大肠杆菌56 株（23.8%）、肺炎克雷伯菌42 株（17.9%）、肠球菌24 株（10.3%）、链球菌13 株（5.6%）。对本试验分离到的大肠杆菌、葡萄球菌、肺炎克雷伯菌等12 种菌中的10 种细菌药敏试验结果显示,不论是革兰氏阳性菌还是革兰氏阴性菌对试验选取的17 种常见抗生素均表现不同程度的耐药,有部分菌株还表现严重的多重耐药,有些菌株对部分药物耐药率达100.00%。