Isolation,Identification and Analysis of Serotype and Antibiotic Susceptibility of Pathogenic Salmonella from Chickens

To investigate the situations of predominant strain and antibiotic resistance of pathogenic Salmonella from chickens in Guangxi Zhuang Autonomous region, Salmonella was pre-enriched and isolated from tissues of clinical suspected chickens, and the Salmonella isolates were identified by biochemical test using ID 32E System for the identification of Enterobacteriaceae of VITEK System ATB Expression, serotypes were determined by slid agglutination test, antibiotic susceptibility was analyzed by ATB VET Susceptibility Test Strip of Enterobacteriaceae antibiotics.The results showed that 34 Salmonella strains were obtained from 310 clinical samples, of which 1 strain belonged to A serogroup, 1 to C2 serogroup, 15 to B serogroup, 14 to D serogroup and 3 to untyped serogroup, and S.typhimurium of B serogroup and S.pullorum of D serogroup were the predominant serotypes.All Salmonella isolates were sensitive to meropenem, imipenem, spectinomycine and apramycine, and the resistance rates to chloramphenicol, kanamycine and gentamicine were less than 10%, and the resistance rates to amoxicillin, streptomycine, flumequine, oxolinique, sulfamethizol, tetracycline and nitrofurantoine ranged between 50% and 90%, while the resistance rates to penicilline, oxacilline, fusidique, rifampicine and metronidazole reached to 100%.The results indicated that the serotype distribution of pathogenic Salmonella from chickens exhibited regional characteristics, and S.typhimurium and S.pullorum were the predominant serotypes in Guangxi Zhuang Autonomous region, and antibiotic resistance of Salmonella isolates was very serious which should be highly concerned.     

Isolation,Identification and Multidrug-resistant Analysis of the New Sequence Type ST989 Strain of Enterococcus faecium

Gram-positive cocci were isolated from half a month diseased lambs with a fatal infection in a sheep farm in Minhe county, Qinghai province.The clinical symptoms observed were drooping, reduced feed intake, emaciated and fever.The infected lambs died several hours after the disease been observed.Necropsy of two dying lambs revealed that for one lamb, the liver was enlarged and capsule of the liver dropped, and other organs had no obvious changes;For the other one, the organs had no obvious changes.After gram staining, gram-positive cocci were observed from all the samples.Partial Tuf gene sequences of these isolates were amplified and sequenced for species identification, BLASTn conparison result revealed that they had the highest sequence similarity with that of Enterococcus faecium (E.faecium) AUS0085 (99.8%), we thus primarily confirmed that these strains belonged to E.faecium.Antimicrobial susceptibility test indicated that these strains could be divided into two classes.Class 1 was resistant to β-lactams, aminoglycosides, macrolides, tetracyclines, quinolones, rifamycins, lincomycin and clindamycin;Susceptible to chloromycetin and vancomycin.Class 2 was resistant to β-lactams, aminoglycosides, tetracyclines, quinolones, rifamycins;Intermediate resistance to erythromycin and roxithromycin;But susceptible to clarithromycin, azithromycin, clindamycin, lincomycin, chloromycetin and vancomycin.Multilocus sequence typing (MLST) based on seven housekeeping genes for E.faecium revealed that E.faecium lby1 strain in class 1 and E.faecium lbg3 strain in class 2 were defined as a new sequence type ST989.Phylogenetic tree analysis revealed the representative strain of these isolates, E.faecium lbg3, was clustered together with E.faecium ST468.The pathogenicity test revealed that E.faecium lby1 and lbg3 strains both had pathogenicity.     

PA-X基因的长度变化对H3N2犬流感病毒复制能力和致病性的影响

H3N2犬流感病毒(canine influenza virus, CIV)已在中国多地的犬群中流行,是禽流感跨宿主感染并形成新分支的近期案例。研究表明,PA-X基因与甲型流感病毒适应新宿主的能力相关,且其长度能够影响甲型流感病毒的复制及致病能力。为了解PA-X基因的长度变化对H3N2 CIV复制能力及致病力的影响,本研究利用H3N2 CIV的8质粒操作系统,拯救了三株重组H3N2 CIV毒株：PA-X基因表达大小为232个氨基酸多肽的亲本病毒CIV＿PA-X＿232;对PA编码区第191、192位氨基酸的密码子进行改造,PA-X基因不表达蛋白的重组病毒CIV＿PA-X＿Knock;对PA+1编码区第232位氨基酸进行突变,PA-X基因表达大小为252个氨基酸多肽的重组病毒CIV＿PA-X＿252。通过比较3株重组病毒的聚合酶活性,在MDCK细胞中的复制效率及对小鼠致病性的差异,来评价表达不同长度的PA-X基因对H3N2 CIV的影响。结果显示,CIV＿PA-X＿252和CIV＿PA-X＿Knock的聚合酶活性显著(P<0.05)高于CIV＿PA-X＿232,且CIV＿PA-X＿...     

‘芽黄’红瑞木炭疽病病原菌的分子鉴定

‘芽黄’红瑞木炭疽病在上海普遍发生。本研究从上海迪士尼园区采集疑似炭疽病的红瑞木病叶,进行了分离培养,获得2种培养形态的炭疽菌,致病性测定证明其为致病菌。联合核糖体转录间隔区、3-磷酸甘油醛脱氢酶、钙调蛋白和β-微管蛋白2多基因序列,使用最大似然法和贝叶斯分析法分别构建了多基因系统进化树。结果显示,分离获得的2株炭疽菌菌株分别与暹罗炭疽菌Colletotrichum siamense和胶孢炭疽菌C.gloeosporioides聚集在一个分支,辅以形态学特征,鉴定引起‘芽黄’红瑞木炭疽病的病原菌为暹罗炭疽菌和胶孢炭疽菌,这是国内关于红瑞木炭疽病的首次报道。     

Phaeosphaeria sp.HD-06菌株对马唐的致病性分析及初步鉴定

为了得到能够有效抑制马唐Digitaria sanguinalis生长的生防微生物,在农田采集马唐种子,进行萌发试验,观察种子的萌发状况,从发病的种子和马唐幼苗基部分离致病菌,通过形态学观察以及ITS序列分析对致病菌进行鉴定。结果从罹病的马唐幼苗基部分离得到一株致病菌HD-06,该菌株的发酵液用水稀释至60%浓度时,对马唐种子的萌发抑制率和幼苗发病率均达到100%,这说明菌株HD-06对马唐有较好的生物防治作用。根据其形态学特点及ITS序列分析结果,鉴定菌株HD-06为暗球腔菌属Phaeosphaeria的真菌。这些结果为深入研究其对马唐的防治作用奠定了基础。     

Potential host range of four Phytophthora interspecific hybrids from Clade 8a

In recent years several interspecific hybrids have been reported in the plant pathogenic oomycete genus Phytophthora. Due to the large genotypic and phenotypic changes, these hybrids might have broader or more limited host ranges compared with their parental species. It is crucial to understand the host range of Phytophthora hybrids to minimize the economic losses caused by their infection. The potential host range of four hybrids belonging to Clade 8a of the Phytophthora phylogenetic tree was investigated in this study. Thirty species of herbaceous plants as well as eight species of woody plants were inoculated and monitored for any symptom of infection. In addition, the detached twigs of 32 tree species, fruits of six plant species, tubers of potato, and roots of carrot and sugar beet were investigated for susceptibility to these hybrids. Almost all hybrids caused severe rot on all tested fruits, tubers, and roots, although different isolates showed different pathogenicity on detached tree twigs. All hybrids tested had a different host range compared with their parental species: they were able to infect plants outside the host range of their parents, infect hosts of both parental species, although these parents did not have overlapping hosts, or, in some cases, they were not able to infect hosts infected by the parents.     

芝麻棒孢叶斑病病原菌鉴定与致病力比较

 为明确芝麻叶斑病病原菌及其致病力,本研究以河南不同地区芝麻上分离所得叶斑病分离物为研究对象,通过形态观察和分子鉴定探讨这些分离物的分类属性,并且进行离体叶片和活体茎杆接菌,进一步探究了不同菌株的致病力差异。结果显示分离所得的6株芝麻叶斑病分离物的菌落形态、生长速率和产孢量存在差异,分生孢子多数呈倒棍棒状,具隔膜,6个菌株的ITS序列与GenBank中Corynespora cassiicola的一致性达99%以上,结合形态特征与ITS序列将其鉴定为山扁豆生棒孢。室内人工接菌芝麻发现,这6个菌株的致病力存在显著的差异;在芝麻叶片上,20180909-03属强致病力菌株,20180821-01、20180909-05和20180824-01属中等致病力菌株,20180904-02-02和SF1-1属弱致病力菌株;在芝麻茎杆上,20180824-01致病力最强,其次是20180821-01、20180909-03和20180909-05,SF1-1和20180904-02-02致病力最弱。研究结果证明山扁豆生棒孢不同菌株在芝麻上的致病力存在明显的分化。     

东北地区水稻纹枯病菌致病性及遗传多样性分析

为明确东北地区水稻纹枯病菌致病性及其群体遗传多样性,将2018年采自东北三省13个水稻主产区的病样分离纯化,共获得176个立枯丝核菌菌株。采用离体叶片接种法测定菌株致病力,利用SRAP分子标记技术分析其遗传多样性。致病力测定结果表明,东北三省各地区菌株致病力存在分化现象,供试的176个菌株中,强致病力菌株占全部菌株23.3%;中等致病力菌株占61.9%;弱致病力菌株占14.8%。地理来源与致病力无明显相关性。根据UPGMA聚类分析,当相似系数为0.63时,176个菌株被划分为6个类群。聚类结果与地理来源具有相关性,但与致病力无明显相关性。利用16对SRAP引物分析176个立枯丝核菌菌株遗传多样性,共得到2 237个扩增条带,多态性频率为82.16%;基因多样度(h)、Shannon信息指数(I)分别为0.2827、0.4150。东北各地区立枯丝核菌群体间遗传距离与地理距离呈一定正相关。群体遗传分化系数Gst=0.3319,基因流Nm=1.0063,说明东北地区立枯丝核菌群体遗传分化度较高且群体间存在基因交流。AMOVA分析结果表明,群体内遗传分化为67.84%,遗传变异主要发生在群体内部。