交叉中和血清1型和血清3型鸭甲肝病毒单克隆抗体的制备与鉴定

鸭甲型肝炎（duck hepatitis A,DHA）是危害养鸭业的主要疾病之一,是一种雏鸭的急性、高度致死性传染病。本研究旨在研制一种对我国流行的鸭甲型肝炎血清1型（DHAV-1）和血清3型（DHAV-3）病毒具有交叉中和作用的单克隆抗体（monoclonal antibody,McAb）。人工合成12条DHAV-1和DHAV-3的VP1蛋白共有的抗原表位肽,分别与钥孔血蓝蛋白（keyhole limpet hemocyanin,KLH）载体偶联作为免疫抗原,免疫BALB/c小鼠制备McAb。通过检测McAb与DHAV-1和DHAV-3的交叉反应性,测定McAb对病毒增殖的抑制效率、中和效价以及攻毒保护率等来筛选McAb。本研究共获得了12株稳定分泌McAb的杂交瘤细胞株,其中6株（C1、C4、C7、C12、C13、C16）分泌的McAb同时与DHAV-1和DHAV-3发生特异性交叉反应; C1、C4、C7、C16可以抑制DHAV-1和DHAV-3在鸭胚中的增殖,抑制率在75.34%～100.00%不等;对DHAV-1和DHAV-3的中和效价：C1（1：3&1：5）、C4（1：3&1：3）、C7（1：10&1：11）和C16（1：9&1：9）;C7和C16对DHAV-1和DHAV-3攻毒雏鸭的保护率较高,分别为“70%、80%”和“100%、60%”。本研究成功研制出对DHAV-1和DHAV-3具有交叉中和活性的McAb 4株,其中2株对病毒的感染具有良好的预防效果,为DHA的防控提供了新材料和新思路。     

广东省养禽场肠球菌耐药性及毒力因子流行分布特征

旨在调查和分析广东省养禽场肠球菌的亚型屎肠球菌和粪肠球菌耐药性及其毒力因子流行分布特征,为控制禽源肠球菌耐药性传播、保障公共卫生安全提供理论依据。作者于2018年从广东省4个养禽场采集肠道样品493份,进行屎肠球菌和粪肠球菌的分离鉴定;采用琼脂二倍稀释法测定肠球菌的最小抑菌浓度（MIC）;PCR方法检测肠球菌的耐药基因和毒力基因。结果显示：1）共分离到125株肠球菌,其中粪肠球菌84株（鸡源66株,鸭源18株）;屎肠球菌41株,均来自鸡肠道样本。2）菌株对四环素、多西环素、红霉素几乎全部耐药,对氟苯尼考和氯霉素的耐药率高达89.60%和74.40%。屎肠球菌耐药率普遍高于粪肠球菌,而粪肠球菌对环丙沙星和利奈唑胺的耐药率高于屎肠球菌;鸭源粪肠球菌对利奈唑胺的耐药率（94%）显著高于鸡源粪肠球菌（39.4%）,屎肠球菌对利奈唑胺均敏感。从鸡分离的1株粪肠球菌对万古霉素耐药。3）耐药基因在屎肠球菌中的检出率高于粪肠球菌,鸭源分离株检出率高于鸡源。耐药基因tetL、fexA、ermB最为流行,检出率均高于90%。其次是optrA基因,检出率为73.60%,poxtA和fexB的检出率均低于20%。在3株鸭源粪肠球菌中检测出cfr基因。4）已检测的毒力基因中efaA的携带率最高,为63.04%（58/92）,其他依次为gelE（54.35%,50/92）、ace（47.83%,44/92）、asa1（44.57%,41/92）。对环丙沙星及高浓度氨基糖苷类耐药的菌株及携带cfr基因的菌株,大多携带agg、asal、gelE或ace。本研究显示养殖场禽源肠球菌耐药严重,鸭源肠球菌对利奈唑胺耐药率高,耐药基因和毒力基因流行且多样,且检测出人医临床重要抗生素耐药基因,应加强对养禽场肠球菌耐药性监测。     

Identification of the core motif of the BRCA2 C-terminal RAD51-binding domain by comparing canine and human BRCA2

Mammary tumors are the most common tumors in women and non-spayed female dogs. One of the reasons for mammary tumors is mutations of the tumor suppressor gene, BRCA2. BRCA2 participates in homologous recombination repair by interacting with the RAD51 recombinase. BRCA2 has two RAD51-binding domains, consisting of BRC repeats and the C-terminal RAD51-binding domain, respectively. Although several studies have addressed the function of the C-terminal RAD51-binding domain of human BRCA2, the amino acid sequences required for the RAD51-interaction activity remain unclear. In this study, the C-terminal RAD51-binding domains of canine and human BRCA2 were compared; the canine domain displayed a weaker interaction with RAD51. This difference was attributed to the C-terminal portion of the domain via a comparison between canine and human domains. Furthermore, peptides shorter than those previously reported displayed RAD51-interacting activity, and a core motif of this domain consisting of 25 amino acids was identified. Since a mutation (S3323N) was reported in the core motif of this domain, the effect of this mutation was evaluated. The mutant exhibited similar RAD51-binding activity as that of the wild-type protein, suggesting that the mutation was functionally neutral. These data suggested that the C-terminal portion of the BRCA2 C-terminal RAD51-binding domain influenced its RAD51-interaction activity, and a minimum core motif of 25 amino acids was identified in this domain. These data may help clarify BRCA2 function, as well as the tumorigenic effects of BRCA2 mutation.     

A survey of antimicrobial resistance in Escherichia coli isolated from wild sika deer (Cervus nippon) in Japan

We examined the antimicrobial susceptibility of 848 Escherichia coli isolates from 237 feces samples of wild sika deer (Cervus nippon) captured between 2016 and 2019 in 39 of the 47 prefectures of Japan. Five of the 237 wild sika deer (2.1%) carried E. coli with resistance to at least one antimicrobial, and all the resistant isolates showed resistance to tetracycline. The resistant isolates contained antimicrobial resistance genes that were similar to those in E. coli derived from humans and farm animals. Although wild sika deer are not currently likely to be a source for the transmission of antimicrobial resistance in Japan, they can potentially mediate antimicrobial resistance spread by coming into contact with humans, animals, and their surroundings.     

Prevalence of antimicrobial-resistant staphylococci in nares and affected sites of pet dogs with superficial pyoderma

Currently, antimicrobial-resistant staphylococci, particularly methicillin-resistant Staphylococcus pseudintermedius (MRSP), are frequently isolated from canine superficial pyoderma in Japan. However, little is known regarding the nasal prevalence of MRSP in pet dogs. Here, we determined the prevalence of antimicrobial-resistant staphylococci in nares and affected sites of pet dogs with superficial pyoderma. Of the 125 nares and 108 affected sites of pet dogs with superficial pyoderma, 107 (13 species) and 110 (eight species) staphylococci strains, respectively, were isolated. The isolation rate of S. pseudintermedius from pyoderma sites (82/110 strains, 74.5%) was significantly higher than that from nares (57/107 strains, 53.3%) (P<0.01). Notably, the prevalence of MRSP (18/57 strains, 31.6%) in nares was equivalent to that in pyoderma sites (28/82 strains, 34.1%). Furthermore, the phenotypes and genotypes of antimicrobial resistance in MRSP strains from nares were similar to those from pyoderma sites. Our findings revealed that the prevalence of antimicrobial-resistant staphylococci in the nares of pet dogs with superficial pyoderma is the same level as that in affected sites. Therefore, considerable attention should be paid to the antimicrobial resistance of commensal staphylococci in companion animals.     

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.     

抗菌肽粗提物对肉鸡生长性能、粪便微生物菌群结构及经济效益的影响

为分析抗菌肽粗提物对肉鸡生长性能、粪便微生物菌群结构及经济效益的影响,试验选择体重相近的21日龄白羽肉鸡600只,随机分成4组,每组15个重复,每个重复10只,1组为空白对照组饲喂基础日粮,试验2、3、4组在基础日粮中分别添加1.0%、1.5%、2.0%的抗菌肽粗提物,试验期为21 d。结果表明：（1）试验2、3、4组的平均采食均高于1组（P > 0.05）|试验3、4组的平均日增重较1组比分别提高11.0%、9.4%（P ＜ 0.05）,料重比较1组比分别降低18.5%、15.2% （P ＜ 0.05）|（2）试验3、4组粪便中大肠杆菌的含量较1组分别降低14.3%、18.0%（P ＜ 0.05）,试验3、4组粪便中乳酸杆菌、双歧杆菌的含量较1组比分别提高4.6%、6.7%、14.5%、12.3%（P ＜ 0.05）|（3）试验3、4组每只肉鸡经济效益较1组比分别提高19.3%、17.7%。综上,1.5%抗菌肽粗提物添加可以提高白羽肉鸡生长性能,改善粪便微生物菌群结构,提高其经济效益。 [关键词] 抗菌肽粗提物|白羽肉鸡|生长性能|粪便微生物菌群结构|经济效益     

酸与碱处理对海带多糖提取及其抗氧化活性的影响

本试验采用柠檬酸提取法和碱提取法从海带中提取多糖组分,以总抗氧化能力和羟自由基（·OH）为衡量抗氧化活性指标,比较这两种提取方法得到海带粗多糖产率和抗氧化活性。试验结果表明：柠檬酸提取法和碱提取法的提取率分别为15.92%和6.05%,所提取的海带粗多糖具有良好的抗氧化活性,且柠檬酸提取法抗氧化活性更高。 [关键词] 柠檬酸提取法|碱提取法|海带多糖|抗氧化活性     

菌丝霉素分泌型丁酸梭菌防治罗非鱼链球菌病的应用研究

为解决罗非鱼链球菌感染难题,本研究在抗菌肽菌丝霉素分泌型丁酸梭菌制剂的基础上,对其进行了抑菌活性、稳定性和罗非鱼养殖应用研究。结果发现:表达产物菌丝霉素对常见致病菌如大肠杆菌、沙门氏菌、金黄色葡萄球菌、蜡样芽孢杆菌和猪链球菌均有较好的抑菌活性。选用米糠作为菌丝霉素分泌型丁酸梭菌的保护剂载体,此制剂可在常温保存一年。通过上述的应用基础试验,初步建立一套菌丝霉素分泌型丁酸梭菌制剂养殖应用方案,此方案可显著降低罗非鱼链球菌病发病率58.6%以上。本研究为丁酸梭菌制剂在防治罗非鱼链球菌病应用方面提供了参考,此制剂有望替代抗生素,促进罗非鱼养殖业的持续健康发展。