Prevalence and characteristics of extended spectrum β-lactamase-producing Escherichia coli from bovine mastitis cases in China

The aim of the study was to investigate the prevalence and characterization of extended-spectrum β-lactamase(ESBL)-producing Escherichia coli isolated from bovine mastitis cases in China. Chrom ID ESBL agar was used to confirm ESBL-producing E. coli. PCR and DNA sequencing were employed to characterize the genotype of ESBL-producers. Antimicrobial susceptibility was measured by disc diffusion. Overall, 73 of 318 E. coli isolates(22.96%) were identified as ESBL-producers. Of these ESBL-producing E. coli, the prevalence of blaCTX-M and bla_(TEM-1) was 97.26 and 71.23%, respectively. The predominant CTX-M-type ESBL was CTX-M-15(65.75%), followed by CTX-M-14(10.96%), CTX-M-55(9.59%), CTX-M-64(5.48%), CTX-M-65(4.11%) and CTX-M-3(1.37%). This study is the first report of CTX-M-64 and CTX-M-65 in E. coli isolated from bovine mastitis. Furthermore, 72 ESBL-producing E. coli isolates(98.63%) were found to be multidrug-resistance. This study noted high prevalence and rates of antimicrobial resistance of ESBL-producing E. coli isolates from bovine mastitis cases in China.     

Detection of antimicrobial resistance and virulence-related genes in Streptococcus uberis and Streptococcus parauberis isolated from clinical bovine mastitis cases in northwestern China

The objectives of this study were to investigate antimicrobial resistance of Streptococcus uberis and Streptococcus parauberis isolated from cows with bovine clinical mastitis in China and to examine the distribution of resistance- and virulence-related gene patterns. Antimicrobial susceptibility was determined by the E-test. Genes encoding antimicrobial resistance and invasiveness factors were examined by PCR. A total of 27 strains were obtained from 326 mastitis milk samples. Streptococcus parauberis isolates (n=11) showed high resistance to erythromycin (90.9%), followed by tetracycline (45.5%), chloramphenicol (36.4%) and clindamycin (27.3%). Streptococcus uberis isolates (n=16) were highly resistant to tetracycline (81.3%) and clindamycin (62.5%). Both species were susceptible to ampicillin. The most prevalent resistance gene in S. uberis was tetM (80.0%), followed by blaZ (62.5%) and ermB (62.5%). However, tetM, blaZ, and ermB genes were only found in 27.3, 45.5, and 27.3%, respectively, of S. parauberis. In addition, all of the isolates carried at least one selected virulence-related gene. The most prevalent virulence-associated gene pattern in the current study was sua+pauA/skc+gapC+hasC detected in 22.2% of the strains. One S. uberis strain carried 7 virulence-associated genes and belonged to the sua+pauA/skc+gapC+cfu+hasA+hasB+hasC pattern. More than 59.3% of analysed strains carried 4 to 7 virulence-related genes. Our findings demonstrated that S. parauberis and S. uberis isolated from clinical bovine mastitis cases in China exhibited diverse molecular ecology, and that the strains were highly resistant to antibiotics commonly used in the dairy cow industry. The data obtained in the current study contribute to a better understanding of the pathogenesis of bacteria in mastitis caused by these pathogens, and the findings are relevant to the development of multivalent vaccines and targeted prevention procedures.     

奶牛乳腺炎源大肠杆菌中耶尔森菌强毒力岛相关基因的检测及序列分析

【目的】研究奶牛乳腺炎源大肠杆菌中耶尔森菌HPI携带情况及其与O血清型的关系,并对部分菌株的相关基因序列进行分析。【方法】从中国北京、内蒙古、甘肃、四川、重庆、云南、贵州等7个省市部分地区1 260份临床型和隐性奶牛乳腺炎奶样中分离得到190株大肠杆菌,对分离菌株进行耶尔森菌强毒力岛核心区irp2基因、fyuA基因及HPI毒力岛在大肠杆菌染色体中插入位置的鉴定,分析HPI毒力岛的携带情况及其与分离菌株O血清型之间的关系。【结果】190株大肠杆菌分离株中,irp2基因阳性率为26.31%（50/190）,fyuA基因阳性率为18.94%（36/190）。50株HPI+分离株中检出asn_tRNA_intB 基因32株,阳性率为64%（32/50）。本试验克隆的irp2基因(273 bp）、fyuA基因(1 071 bp）、asn_tRNA_intB基因(1 512 bp）均与已发表序列高度同源,同源性分别在97.1%、98.2%、97.2%以上,且其HPI毒力岛大多位于大肠杆菌染色体的asn_tRNA位点上。【结论】耶尔森菌HPI在奶牛乳腺炎源大肠杆菌中广泛流行分布,但也存在差异,而不同血清型菌株携带HPI的倾向性可能只与特定的血清型有一定的关系。     

Month-Wise Prevalence of Subclinical Mastitis in Dairy Cows in Guangdong Province,China

To evaluate the month-wise prevalence of subclinical mastitis (SM) and its relations with climatic temperature and humidity, and to provide references for control and prevention of mastitis in dairy cows in Guangdong, China, California mastitis test method was used to determine the monthly prevalence of SM (MPSM, %) of lactating Holstein cows from 11 dairy farms in Guangdong Province, China, including about 516 cows per monthly examination. The average MPSM on a cow and a quarter basis were 30 and 13% respectively, and there was a positive linear correlation between monthly mean air temperature (MT, ?C) and MPSM on a cow basis (r=0.763, P=0.004) or a quarter basis (r=0.577, P=0.049), but there were no correlations between MT and MPSM on a cow or a quarter basis (P>0.05). There was a shift in MPSM trend that the highest MPSM (38.4%) was not for the hottest July with MT at 30.8?C, but for the later September at 27.9?C. The farms need to develop new housing conditions to control cowshed temperatures in the hot season to reduce the mastitis prevalences in the posthot months.     

Estimation of the potential geographical distribution of a new potato pest (Schrankia costaestrigalis) in China under climate change

Global food security is threatened by the impacts of the spread of crop pests and changes in the complex interactions between crops and pests under climate change. Schrankia costaestrigalis is a newly-reported potato pest in southern China. Early-warning monitoring of this insect pest could protect domestic agriculture as it has already caused regional yield reduction and/or quality decline in potato production. Our research aimed to confirm the potential geographical distributions (PGDs) of S. costaestrigalis in China under different climate scenarios using an optimal MaxEnt model, and to provide baseline data for preventing agricultural damage by S. costaestrigalis. Our findings indicated that the accuracy of the optimal MaxEnt model was better than the default-setting model, and the minimum temperature of the coldest month, precipitation of the driest month, precipitation of the coldest quarter, and the human influence index were the variables significantly affecting the PGDs of S. costaestrigalis. The highly- and moderately-suitable habitats of S. costaestrigalis were mainly located in eastern and southern China. The PGDs of S. costaestrigalis in China will decrease under climate change. The conversion of the highly- to moderately-suitable habitat will also be significant under climate change. The centroid of the suitable habitat area of S. costaestrigalis under the current climate showed a general tendency to move northeast and to the middle-high latitudes in the 2030s. The agricultural practice of plastic film mulching in potato fields will provide a favorable microclimate for S. costaestrigalis in the suitable areas. More attention should be paid to the early warning and monitoring of S. costaestrigalis in order to prevent its further spread in the main areas in China’s winter potato planting regions.     

Neopestalotiopsis eucalypti,a causal agent of grapevine shoot rot in cutting nurseries in China

Grapevine (Vitis vinifera L.) is an economically important fruit crop in the world, and China ranks first in the production of grapes with approximately 15% of the world's total yield. However, diseases that cause the death of grapevine shoots pose a severe threat to the production of grapes. In this study, the fungus Neopestalotiopsis eucalypti was identified as a causal pathogen of grapevine shoot rot based on the morphology of conidia and a phylogenetic analysis. The phylogenetic analysis was performed with three isolates based on the combined sequence of internal transcribed spacer (ITS) region of ribosomal DNA, part of the translation elongation factor 1-alpha (Tef) and the β-tubulin (Tub2) genes. The three isolates were all identified as N. eucalypti. Pathogenicity tests of the three fungal isolates were conducted on grapevines shoots in vitro and in vivo. The results showed that all three fungal isolates caused severe rot lesions on the inoculated grapevine shoots, and N. eucalypti was re-isolated from the inoculated grapevine shoots. Therefore, N. eucalypti was confirmed as a causal agent of the grapevine shoot rot. This is the first report of N. eucalypti causing grapevine shoot disease in China.     

奶牛乳腺炎葡萄球菌生物被膜形成及相关基因分析

 【目的】研究临床分离的奶牛乳腺炎葡萄球菌生物被膜形成能力,相关基因的分布及二者之间的关系;【方法】利用通用硅胶片法培养葡萄球菌生物被膜,经充分洗涤后对生物被膜进行银染定性判断菌株形成生物被膜的能力,通过生物被膜菌落结晶紫染色法定量检测菌株生物被膜的形成能力,并用扫描电镜观察被膜结构,对葡萄球菌bap、icaAD、icaBC、sar、agr、sigB、clfaA、clfaB、fnbpA和fnbpB进行PCR扩增检测。【结果】 银染法定性结果显示137株葡萄球菌中有120株形成肉眼可见的生物被膜,生物被膜形成率为87.6%;结晶紫染色定量检测与硅胶粘附的有132株,不粘附的有5株。试验所用的137株葡萄球菌中,57株能扩增出bap基因;有43株和54株分别能扩增出icaAD和icaBC;有73株、49株和38株分别扩增出sigB、sar和agr;分别有76株和50株染色体中存在clfaA和clfaB;fnbpA和fnbpB分别在52株和26株菌株中扩增出。【结论】推测bap、sigB、sar、icaAD和icaBC基因是生物被膜形成的重要相关基因;agr及粘附素基因clfaA、clfaB、fnbpA和fnbpB对生物被膜形成的作用不明显。     

Characterization of blaCTX-M Gene in One Klebsiella pneumoniae Isolate from Sick Chickens in China

Two Klebsiella pneumoniae isolates (Kpc1 and Kpc2) were obtained from liver samples of seven dead chickens and identified with Vitek-32 automated identification system. Antimicrobial susceptibilities were determined by the microdilution broth method. Detection of genes encoding class A β-lactamases was performed by PCR amplification, and cloning of the ESBL gene was by plasmid restriction and fragments ligation. Conjugation assay, transformation experiments and plasmid profile analysis were performed. The incompatibility group of ESBL-carrying plasmid was determined by the PCR-based replicon typing method. Lastly, the genetic environment was analysed by direct sequencing of the DNA surrounding the ESBL gene. The genes associated with tetracycline and gentamicin resistance were also sought by PCR. The results revealed that the ESBL phenotype-negative strain Kpc2 only showed resistance to ampicillin, amoxicillin, tetracycline, and doxycycline and carried bla_TEM-1 and tet(A) genes. The ESBL-producing strain Kpc1 exhibited multidrug resistant phenotype and harbored bla_TEM-1, bla_CTX-M-14, tet(A), tet(B), and rmtB genes. K. pneumoniae Kpc1 contained four plasmids with molecular sizes of approximately 59, 6.9, 2.8, and 1.6 kb, but only a 59-kb plasmid, carried bla_TEM-1 and bla_CTX-M-14 genes, was observed in its transconjugant. The incompatibility group of plasmid carrying bla_CTX-M-14 gene could not be determined. The bla_CTX-M-14 gene was flanked upstream by an ISEcp1 insertion sequence and downstream by an IS903 element. This work shows that CTX-M-14 is present in K. pneumoniae isolates from chickens in China. The bla_CTX-M-14 gene was associated with an upstream ISEcp1 insertion sequence. Our results underline the need for continuous surveillance of the prevalence and evolution of this CTX-M-type β-lactamase in China.     

广东省鸭疫里默氏杆菌流行病学监测及遗传进化关系

【目的】明确广东地区鸭疫里默杆菌Riemerella anatipestifer的血清型、耐药状况及遗传进化关系。【方法】从规模化鸭场分离鉴定鸭疫里默氏杆菌,通过玻片凝集试验鉴定血清型;利用试管两倍稀释法测试抗菌药物的最低抑菌浓度,分析药物的敏感性;采用全基因组测序技术分析序列特征并构建核心基因组遗传进化树。【结果】共分离鉴定鸭疫里默氏杆菌168株,血清1、2、3、5、6、7、8、10型均有流行,血清1型的菌株高达54.17%(91/168),其次为2型,占27.97%(47/168)。48株代表性菌株对庆大霉素、卡那霉素、盐酸环丙沙星表现高度耐药,耐药率均超过80%;对土霉素、盐酸四环素、盐酸金霉素、氧氟沙星、诺氟沙星、磺胺二甲嘧啶、磺胺对甲氧嘧啶的耐药率均在60%以上;对阿莫西林、头孢噻肟和大观霉素的耐药率低于30%。受试菌株对5～12种药物耐药,共有44种耐药谱型。成功获得46株菌株全基因组序列,共检出6种耐药基因,其中,耐药基因erm(F)和tet(X)的检出率较高,分别为73.91%(34/46)和82.60%(38/46),同时携带2种以上耐药基因的菌株占95.65%(44...     

Genomic characteristics of Dickeya fangzhongdai isolates from pear and the function of type IV pili in the chromosome

Dickeya fangzhongdai, the causal agent of bleeding canker of pear, is a new member of the Dickeya genus and the only one that infects woody plants. Recent studies have reclassified several Dickeya isolates as D. fangzhongdai, which were isolated from various environments, including water, Phalaenopsis sp. and Aglaonema sp. To provide genomic characterization of D. fangzhongdai isolates from pear, the genomes of D. fangzhongdai strain JS5 (=China General Microbiological Culture Collection Center, CGMCC 1.15464^T=DSM 101947^T), along with two other isolates, LN1 and QZH3, were sequenced and compared to those of other Dickeya spp. Homology greater than 99% was observed among three D. fangzhongdai strains. Plasmid, type IV secretion system (T4SS) and type IV pili (TFPs) were found in genomes of D. fangzhongdai isolates. Comparative analysis of the type III secretion systems (T3SS), type III secretion effectors (T3SE), plant cell wall degradation enzymes (PCWDE) and membrane transport proteins of Dickeya spp. showed some differences which might reflect the variations of virulence, phylogenetic and phenotypic characteristics of Dickeya spp. In addition, deletion mutant of TFP in D. fangzhongdai JS5 showed no twitching motility and reduced virulence and biofilm formation. The fingdings of the distinctive plasmid, T4SS and TFPs, as well as the differences of T3SE, PCWDE and membrane transport proteins make D. fangzhongdai isolates unique. These results also suggested that acquisition of virulence genes by horizontal gene transfer might play some role in the genetic variation of D. fangzhongdai.     

I群禽腺病毒血清4型广东株的分离鉴定及分子特性

[目的]对广东地区疑似心包积液–肝炎综合征患病鸡进行腺病毒分离鉴定并研究其分子特征.[方法]采集的病料样品,通过鸡胚有限稀释法经SPF鸡胚传代分离纯化病毒,进行致病性试验,并对病毒的hexon、fiber-2、ORF19和ORF29基因进行PCR扩增,分析其分子特征.[结果]hexon基因序列分析显示此次分离的毒株为Ⅰ群禽腺病毒血清4型(命名为WM-XC株),SPF鸡接种分离毒株后,致死率达90%,死亡鸡只出现明显的心包积液、肝炎等变化,肝脏病理切片可观察到明显的嗜酸性包涵体,显示分离毒株具有致病性.fiber-2基因序列同经典毒株ON1和KR5相应序列的相似性达96.1%和97.0%,与2015年以来国内分离的毒株序列相似性则达到100%;ORF29序列与2013年国内分离到的毒株JSJ13和JN13的相应序列相比存在33个碱基的缺失;WM-XC分离株与2015年以来国内分离毒株序列相同,但相比经典株ON1和KR-5,缺失ORF19序列.[结论]此次从广东地区分离到的血清4型Ⅰ群腺病毒,和近年国内其他地区分离的流行毒株一样,其fiber-2、ORF19和ORF29基因序列与经典毒株差异明显.     

牛乳源大肠杆菌生物被膜形成及抗生素的作用

为研究大肠杆菌生物被膜（Biofilms，BF）对耐药性的影响及抗生素对BF形成的作用，以陕西关中地区乳房炎奶牛的乳样中分离的 35 株大肠杆菌为研究对象，通过刚果红琼脂和结晶紫染色法测定菌体形成 BF 的能力；通过药物敏感性试验检测 BF 形成前后菌体耐药性的变化；用结晶紫染色法结合荧光显微镜观察不同质量浓度抗生素对 BF 的影响。结果显示，35 株大肠杆菌中有 19 株（54.29%）为 BF 阳性，以形成强、中膜能力为主（84.21%）；大肠杆菌形成BF后使头孢噻呋、链霉素、四环素、环丙沙星、庆大霉素、青霉素的最小抑菌质量浓度及最小杀菌质量浓度分别提高4~16 倍和8~64 倍；抗生素对BF形成的抑制和清除作用随着药物质量浓度增大而加强，其中环丙沙星、头孢噻呋的效果较好。     

亚热带养殖海湾皱瘤海鞘生物沉积的现场研究

皱瘤海鞘是亚热带海域分布最广和数量最多的附着生物种类之一.2012年1月-7月在典型的亚热带养殖海湾-大亚湾大鹏澳海域,利用沉积物捕集器现场测定了皱瘤海鞘(Styela plicata)的生物沉积速率,并测定了生物沉积物中有机物(OM)、总碳(TC)、总氮(TN)、有机碳(OC)和有机氮(ON)含量.结果显示:皱瘤海鞘的生物沉积速率变化范围为每天145.5-1011.8m/个,平均每天516.0mg/个,海鞘的生物沉积速率变化范围为每天154.8-1065.8 mg/g干重,平均每大463.3 mg/g干重.海鞘生物沉积物中OM、TC、OC、TN和ON含量分别为14.38％、10.80％、2.87％、3.06％和0.86％,高于自然沉积物中的含量,分别为13.39％、7.36％、2.32％、2.29％和0.67％.其中TC和ON含量要显著高于自然沉积物(P＜0.05).皱瘤海鞘的OM、TC、OC、TN和ON的生物沉积速率分别为每天74.20,55.73,14.80,15.79和4.43 mg/个.实验期间附着在浮筏养殖设施和养殖牡蛎壳上的海鞘密度变化范围为54.9-222.1个/m2,平均147.5个/m2,养殖海域单位面积的海鞘生物沉积速率平均为每天76.1 g/m2,是自然沉积速率(平均每天62.7 g/m2)的1.21倍,其中OM、TC、OC、TN和ON的平均沉积速率分别为每天10.94,8.21,2.18,2.32和0.65 g/m2.据此可推算,大鹏澳筏式牡蛎养殖海区(约103 hm2)皱瘤海鞘的年生物沉积物负荷为29000 t,其中OM,TC,OC,TN和ON分别4100,3100,820,870和240 t.研究结果说明,海鞘等附着生物在大规模浅海贝类养殖中对养殖生态环境的影响也不容忽视.     

黄瓜种传镰刀菌种类的鉴定及其致病性研究

为明确我国主栽黄瓜品种的种子中携带镰刀菌的种类及其危害,对来自我国黄瓜主产区21个黄瓜品种的种子进行种传镰刀菌检测,从种胚和种子外部分离得到镰刀菌分离物9个,采用形态学及分子生物学的方法进行鉴定,并研究其对黄瓜种子发芽和幼苗致病性的影响.结果表明:9个分离物中,4个分离物为尖孢镰刀菌(Fusarium oxysporum),4个分离物为串珠镰刀菌(F.moniliforme),1个分离物为再育镰刀菌(F.proli feratum).4个尖孢镰刀菌分离物对黄瓜种子发芽均有显著影响,发芽指数、活力指数、根长和鲜重等指标均显著降低,且能导致黄瓜幼苗出现典型的枯萎症状,经柯赫氏法则检验证明其具有致病性.其他分离物对黄瓜种子发芽也有一定的抑制作用,但没有致病性.     

Genetic Diversity of Chinese Soybean mosaic virus Strains and Their Relationships with Other Plant Potyviruses Based on P3 Gene Sequences

Soybean mosaic virus （SMV）, a member of the genus Potyvirus, is a major pathogen of soybean plants in China, and 16 SMV strains have been identified nationwide based on a former detailed SMV classification system. As the P3 gene is thought to be involved in viral replication, systemic infection, pathogenicity, and overcoming resistance, knowledge of the P3 gene sequences of SMV and other potyviruses would be useful in efforts to know the genetic relationships among them and control the disease. P3 gene sequences were obtained from representative isolates of the above-mentioned 16 SMV strains and were compared with other SMV strains and 16 Potyvirus species from the National Center for Biotechnology GenBank database. The P3 genes from the 16 SMV isolates are composed of 1041 nucleotides, encoding 347 amino acids, and share 90.7-100% nucleotide （NT） sequence identities and 95.1-100% amino acid （AA） sequence identities. The P3 coding regions of the 16 SMV isolates share high identities （92.4-98.9% NT and 96.0-100% AA） with the reported Korean isolates, followed by the USA isolates （88.5-97.9% NT and 91.4-98.6% AA）, and share low identities （80.5-85.2% NT and 82.1-84.7% AA） with the reported HZ 1 and P isolates from Pinellia ternata. The sequence identities of the P3 genes between SMV and the 16 potyviruses varied from 44.4 to 81.9% in the NT sequences and from 21.4 to 85.3% in the AA sequences, respectively. Among them, SMV was closely related to Watermelon mosaic virus （WMV）, with 76.0-81.9% NT and 77.5-85.3% AA identities. In addition, the SMV isolates and potyvirus species were clustered into six distinct groups. All the SMV strains isolated from soybean were clustered in Group I, and the remaining species were clustered in other groups. A multiple sequence alignment analysis of the C-terminal regions indicated that the P3 genes within a species were highly conserved, whereas those among species were relatively variable.     

Seedling and adult plant resistance to leaf rust in 46 Chinese bread wheat landraces and 39 wheat lines with known Lr genes

Wheat leaf rust,caused by Puccinia triticina(Pt),is an important foliar disease that has an important influence on wheat yield.The most economic,safe and effective way to control the disease is growing resistant cultivars.In the present study,a total of 46 wheat landraces and 34 wheat lines with known Lr(leaf rust resistance)genes were inoculated with 16Pt pathotypes for postulating seedling resistance gene(s)in the greenhouse.These cultivars and five wheat differential lines with adult plant resistance(APR)genes(Lr12,Lr22b,Lr34,Lr35 and Lr37)were also evaluated for identification of slow rusting resistance in the field trials in Baoding,Hebei Province of China in the 2014–2015 and 2015–2016 cropping seasons.Furthermore,10 functional molecular markers closely linked to 10 known Lr genes were used to detect all the wheat genotypes.Results showed that most of the landraces were susceptible to most of the Pt pathotypes at seedling stage.Nonetheless,Lr1 was detected only in Hongtangliangmai.The field experimental test of the two environments showed that 38 landraces showed slow rusting resistance.Seven cultivars possessed Lr34 but none of the landraces contained Lr37 and Lr46.Lr genes namely,Lr9,Lr19,Lr24,Lr28,Lr29,Lr47,Lr51 and Lr53 were effective at the whole plant stage.Lr18,Lr36 and Lr45 had lost resistance to part of pathotypes at the seedling stage but showed high resistance at the adult plant stage.Lr34 as a slowing rusting gene showed good resistance in the field.Four race-specific APR genes Lr12,Lr13,Lr35 and Lr37 conferred good resistance in the field experiments.Seven race-specific genes,Lr2b,Lr2c,Lr11,Lr16,Lr26,Lr33 and LrB had lost resistance.The 38 landraces showed slow rusting resistance to wheat leaf rust can be used as resistance resources for wheat resistance breeding in China.     

西藏那曲牦牛源多杀巴氏杆菌荚膜分型及其毒力基因检测

为确定2018年7—11月西藏那曲牦牛发病的死亡原因,以安多县、班戈县、聂荣县和色尼区的26头病死牦牛内脏组织为研究对象,利用常规细菌培养法和特异性基因PCR方法,对分离菌进行鉴定,并对其荚膜血清类型、毒力基因以及致病性进行研究。结果表明:纯化得到13株分离菌,分离菌的菌落特征、菌体形态、生化鉴定结果均与多杀巴氏杆菌相符,并扩增出多杀巴氏杆菌特异性基因Kmt-1和16S rDNA,菌株Pm-1和Pm-2的Kmt-1基因序列与印度、伊朗、巴基斯坦、俄罗斯的牛源多杀巴氏杆菌的同源性均达98%以上;13株菌的荚膜分型鉴定结果均为B型;对分离菌进行已知的23种毒力基因检测结果显示,此次分离到的13株菌毒力基因的数量在16～19种,ptfA、fimA等16种毒力基因检出率为100%,tadD、toxA等4种毒力基因检出率为0,且每个地区分离株毒力基因的分布相同;致病性试验中对照组小鼠全部存活,处理组小鼠死亡时间为24～72 h,死亡率为100%,病变主要表现为脾脏、肺脏、肝脏、心脏出血坏死。导致此次那曲地区牦牛发病死亡的原因为荚膜B型多杀巴氏杆菌病,本研究可为该病的防治提供依据,也为下一步预防疫苗的研究提供了优势菌株。     

Pathotypes and Genetic Diversity of Chinese Collections of Elsinoë fawcettii Causing Citrus Scab

Two scab diseases are currently recognized on citrus: citrus scab, caused by Elsinoë fawcettii, and sweet orange scab, caused by E. australis. Although these pathogens are economically important, there is no molecular data on these species in China. Here we use internal transcribed spacer sequence data to report on host-specificity and genetic relationships among 46 isolates collected from the main citrus varieties grown across China. All strains isolated were E. fawcettii. Based on pathogenicity testing on 9 different citrus species, isolates were divided into 11 pathotypes (SM, FBHR, SJCR, SPOJCR, SR, SOJG, SPOJC, SRGC, Lemon and two unnamed pathotypes). SM is a new pathotype, and two isolates did not fit into any of the known pathotypes of E. fawcettii. Inter-simple sequence repeat (ISSR-PCR) assays separated the E. fawcettii isolates into 10 subgroups; the groupings basically corresponded to the pathogenicity test.     

鸽源大肠杆菌O157：非H7菌株的致病性和耐药性分析

为查明南京市某赛鸽公棚赛鸽出现腹泻、死亡的致病病原,本研究对采集的病料进行病原分离鉴定,分析致病菌的血清型、生物被膜表型、毒力基因、耐药菌谱,并对分离菌株进行了人工感染试验。研究表明:1)从患病鸽组织、拭子和粪便中分离出的细菌初步诊断为大肠杆菌O157;分离菌株在NA平板上长出白色、光滑的圆形菌落,在CT-SMAC平板上长出紫红色菌落。2)分离菌株的16S rRNA基因序列与大肠杆菌的相似性达98%~99%,命名为E.c.86246。3)血清学试验表明分离菌株为大肠杆菌O157:非H7菌株,动力试验进一步证实其为动力株。4)分离菌株携带tccp1014、eaeA和hly毒力基因,可导致雏鸡胃肠道出现不同程度的功能损伤,甚至导致死亡;其半数致死量为2.0×10⁵ CFU/mL,具有较强的致病性。5)分离菌株携带tetA、tetM、sul2、aadA1和bla_TEM 5种耐药基因,具有较弱的生物被膜形成能力,对四环素、氨基糖苷类等抗菌药物严重耐药,对阿莫西林、氧氟沙星和诺氟沙星敏感。综上,本研究分离的鸽源大肠杆菌O157:非H7菌株,具有动力性,有较强的致病性和一定的耐药性,为大肠杆菌O157的流行病学调查、致病机理分析和抗菌药物的精准施治提供理论依据和数据参考。     

Development of a high-efficiency sex pheromone formula to control Euproctis pseudoconspersa

The tea tussock moth (Euproctis pseudoconspersa) is one of the most destructive chewing pests in tea plantations and causes a serious allergic reaction on the skin of tea plantation workers. The sex pheromone components of its Japanese population were first identified as 10,14-dimethylpentadecyl isobutyrate (10Me14Me-15:iBu) and 14-methylpentadecyl isobutyrate (14Me-15:iBu), with a stereogenic center. Only 10Me14Me-15:iBu has been identified in the Chinese E. pseudoconspersa population. However, field tests have shown that 10Me14Me-15:iBu cannot meet the demand of effective pest control in China. To develop a high-efficiency E. pseudoconspersa sex pheromone formula, electroantennogram (EAG) recordings of (S)- and (R)-enantiomers of 10Me14Me-15:iBu and 14Me-15:iBu were obtained in the present study. The results demonstrated that the EAG responses of male antennae to (R)-enantiomers were significantly higher than responses to the (S)-enantiomers, and 14Me-15:iBu also elicited EAG activity. Field tests showed that the catch numbers of male moths by (R)-enantiomers were significantly higher (P<0.05) than those of (S)-enantiomers. Addition of 14Me-15:iBu significantly increased the catch numbers of both the (S)- and (R)-enantiomers. The efficient pheromone formula containing 0.75 mg (R)-10Me14Me-15:iBu and 0.1 mg 14Me-15:iBu showed significantly higher attractiveness than commercial pheromone products. Our study demonstrated that (R)-10Me14Me-15:iBu was the major sex pheromone component of E. pseudoconspersa, and 14Me-15:iBu might be the minor sex pheromone component. Furthermore, a high-efficiency sex pheromone formula for E. pseudoconspersa control was defined in this study.