益生菌大肠杆菌Nissle 1917抗逆性能、猪肠上皮细胞黏附率及抑菌效果研究

本试验旨在研究益生菌大肠杆菌Nissle 1917(Ec N)抗逆性能、猪肠上皮细胞黏附率及抑菌效果。采用体外法对Ec N进行生长曲线绘制和耐酸、耐胆盐、耐热性能的测定;以猪肠上皮细胞IPEC-J2细胞为体外细胞模型,考察了Ec N对该细胞的黏附率以及对致病菌大肠杆菌K88的黏附抑制率;同时通过蛋白质印迹法检测了Ec N对IPEC-J2细胞β-防御素-2和Toll样受体4的水平的影响。结果表明:1)Ec N对高酸、高胆盐和高温环境具有一定耐受能力。2)Ec N对IPEC-J2细胞的黏附作用以对数期最佳,黏附率达33.96%,显著高于迟缓期、稳定期和衰亡期(P0.05)。3)Ec N对致病菌大肠杆菌K88具有良好的抑制效果,黏附抑制率达87.84%。4)Ec N还能上调IPEC-J2细胞β-防御素-2和Toll样受体4水平。结果提示,益生菌Ec N具有较好的抗逆性能,能够良好地黏附猪肠上皮细胞,对致病菌大肠杆菌K88具有良好的抑制作用。     

大蒜素对番茄叶霉菌不同生理小种的抑制作用

研究大蒜素对番茄生产上叶霉菌主流生理小种1.2.3.4以及2014年新发现生理小种2.5、2.4.5和1.2.3.4.9的抑制作用,为开发安全、高效的植物源农药奠定基础。首先制备大蒜素抽提母液,采用定硫法对其中的大蒜素进行定量测定,发现当溶液中滴加4 m L浓硝酸、溶液p H值为2.0时能够更为准确地测得大蒜素的含量,试验测得大蒜素含量为0.52%。在此基础上,进行了大蒜素对4个叶霉菌生理小种菌丝生长、孢子萌发的抑制试验,离体叶片上的病害预防试验以及苗期药效试验。结果表明,当大蒜素剂量25μL/m L时,其对叶霉菌生理小种菌丝生长的抑制活性较高,其中对1.2.3.4的抑制率最高,对1.2.3.4.9的抑制率最低,对其他2个生理小种的抑制率差异不显著。当大蒜素剂量7.5μL/m L时,其对4个生理小种孢子萌发的抑制效应存在差异,其中对1.2.3.4.9的孢子萌发抑制率最低,对1.2.3.4的孢子萌发抑制率最高。当大蒜素剂量为125μL/m L时,其对叶霉菌4个生理小种在离体叶片上的预防效果均超过95%。苗期药效试验表明,在相等剂量下,大蒜素对叶霉菌的防治效果高于多菌灵和甲基托布津,说明大蒜素是一种可以替代化学药剂防治叶霉病的有效抑菌物质。     

艾纳香内生真菌抗细菌和炭疽菌的活性研究

为获得丰富的艾纳香内生真菌资源,筛选出抗性生防菌株,用于活性次生代谢产物的发现,通过水琼脂法对海南产艾纳香进行内生真菌分离,通过ITS序列分析,对其进行鉴定和分析。采用滤纸片法评价内生真菌发酵产物对细菌的抑制活性,采用平板对峙培养法评价艾纳香内生真菌对植物炭疽菌的拮抗作用。本研究共分离鉴定出191株内生真菌,分属于27属,45种,其中优势菌属为Diaporthe、Paraphoma、Ectophoma、Penicillium、Talaromyces、Hypoxylon、Phomopsis、Colletotrichum和Fusarium。抗细菌活性结果显示,Clonostachys、Fusarium和Diapothe属活性较强。抗炭疽病菌试验显示,Talaromyces、Ectophoma和Penicillium属活性较强。本研究初步探讨了海南产艾纳香内生真菌种类的多样性,并首次对其进行了抗病原菌活性筛查,获得的活性菌株可作为微生物源菌剂开发的重要材料。     

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。     

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.     

2019年湖南省部分地区鸡、鸭、猪源大肠杆菌耐药性调查

旨在调查2019年湖南省部分地区不同来源的大肠杆菌对抗生素的耐药水平,为养殖合理用药提供参考。从4个市畜禽养殖场采集猪、鸡和鸭粪便285份,使用麦康凯琼脂培养基和伊红美蓝培养基对大肠杆菌筛选,利用基质辅助激光解析电离飞行时间质谱进行鉴定,采用微量肉汤稀释法检测16种抗生素对大肠杆菌的最小抑菌浓度(MIC)。共分离出203株大肠杆菌(鸡源146株、鸭源20株和猪源37株),对四环素和氨苄西林耐药率最高,分别达到89.2%和84.7%;对头孢他啶、奥格门丁、黏菌素耐药率较低,分别为2.5%、2.0%和0.5%;所有菌株均对美罗培南敏感。抗3种及以上抗生素的菌株数目占比为87.7%,而仅5.42%的菌株对所有检测的抗生素敏感。另外,来源于减抗示范养殖场的大肠杆菌对大部分抗生素的耐药率,与来源于非减抗养殖场大肠杆菌相比未见显著差异。上述结果表明,湖南省动物源大肠杆菌的耐药水平仍处在较高水平,重视兽药使用管理并持续开展对大肠杆菌的耐药性监测十分必要。     

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.     

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     

Isolation and characterization of antimicrobial-resistant Escherichia coli from national horse racetracks and private horse-riding courses in Korea

Limited information is available regarding horse-associated antimicrobial resistant (AR) Escherichia (E.) coli. This study was designed to evaluate the frequency and characterize the pattern of AR E. coli from healthy horse-associated samples. A total of 143 E. coli (4.6%) were isolated from 3,078 samples collected from three national racetracks and 14 private horse-riding courses in Korea. Thirty of the E. coli isolates (21%) showed antimicrobial resistance to at least one antimicrobial agent, and four of the AR E. coli (13.3%) were defined as multi-drug resistance. Most of the AR E. coli harbored AR genes corresponding to their antimicrobial resistance phenotypes. Four of the AR E. coli carried class 1 integrase gene (intI1), a gene associated with multi-drug resistance. Pulsed-field gel electrophoretic analysis showed no genetic relatedness among AR E. coli isolated from different facilities; however, cross-transmissions between horses or horses and environments were detected in two facilities. Although cross-transmission of AR E. coli in horses and their environments was generally low, our study suggests a risk of transmission of AR bacteria between horses and humans. Further studies are needed to evaluate the risk of possible transmission of horse-associated AR bacteria to human communities through horse riders and horse-care workers.