脉冲场凝胶电泳在沙门氏菌流行病学中的应用

脉冲场凝胶电泳是20世纪80年代发展起来的一种可用于分离10 kb至10 Mb分子质量DNA的新型凝胶电泳技术,已广泛应用于细菌分子分型、种群特异性鉴定及遗传分析等方面。文章介绍了脉冲场凝胶电泳的技术、应用状况及目前沙门氏菌的分型现状,并阐述了该技术在沙门氏菌流行病学研究方面的应用及发展前景。     

饲喂乳杆菌雏鸡对鸡白痢菌感染的影响

雏鸡饲喂乳杆菌培养物后,人工感染鸡白痢沙门氏菌,观察雏鸡盲肠主要正常菌群定植及雏鸡抵抗力情况。研究结果表明：人工感染鸡白痢沙门氏菌后,饲喂乳杆菌组同未饲喂乳杆菌组相比,雏鸡盲肠内正常菌群有一定变化,大肠杆菌数明显降低（Ｐ＜００１）,双歧杆菌和乳杆菌数明显增加（Ｐ＜００１）,而且死亡率降低２０％。     

Herd prevalence of Salmonella enterica infections in Danish slaughter pigs determined by microbiological testing

As a part of a nationwide programme to survey and control salmonella in pig herds, a microbiological survey of 1363 pig herds was performed in Denmark. A total of 13 468 slaughter pigs were examined at slaughter by culture of 5 g of caecal contents. Overall, 30 different serotypes of Salmonella enterica were isolated from 832 pigs (6.2%). The predominant serotype was S. Typhimurium, comprising 536 (64.4%) of the isolates. Four hundred and forty-eight isolates of S. Typhimurium were examined by phage typing, resulting in detection of 17 different phage types (definitive types, DT) with DT12 being the most frequent (49.1%).

Salmonella enterica was found in 302 herds (22.2%), S. Typhimurium was found in 61.1% of these. 279 (23.1%) large herds (producing more than 2600 slaughter pigs per year) were found to be salmonella positive compared with 23 (14.7 %) small herds (annual production of 500 to 550 slaughter pigs). Practical constraints in the study design did not allow for a firm conclusion on the interplay among herd size, geographical location and occurrence of salmonella.

In 284 of 302 infected herds (94.0%) only one serotype was detected. Infections with two different serovars were seen in 18 herds (6.0%).     

鸡白痢沙门氏菌与鸡伤寒沙门氏菌基因组消减文库的构建

为了解鸡白痢沙门氏菌与鸡伤寒沙门氏菌的基因组差别,筛选鸡白痢沙门氏菌特有的序列,利用SSH对鸡白痢沙门氏菌C79-13（实验方）与鸡伤寒沙门氏菌Sg9（驱动方）基因组进行了比较。选择四碱基内切酶Rsa I将C79-13与Sg9基因组DNA酶切,酶切的C79-13基因组DNA分成两份,分别与接头1和接头2R连接,然后进行两轮杂交和两轮PCR ,得到消减混合物, 将其与PCR?2.1克隆载体连接,转化感受态E.coli DH5α建立消减文库。结果共获得400个阳性克隆;筛选240个克隆制备质粒进行PCR检测,均扩增出100～2000 bp大小的片段。差异DNA消减文库的构建为进一步筛选、克隆鸡白痢沙门氏菌特异的未知新基因、建立分子鉴别新体系奠定了基础。     

Campylobacter spp., Salmonella spp., Verocytotoxic Escherichia coli,and Antibiotic Resistance in Indicator Organisms in Wild Cervids

Faecal samples were collected, as part of the National Health Surveillance Program for Cervids (HOP) in Norway, from wild red deer, roe deer, moose and reindeer during ordinary hunting seasons from 2001 to 2003. Samples from a total of 618 animals were examined for verocytotoxic E. coli (VTEC); 611 animals for Salmonella and 324 animals for Campylobacter. A total of 50 samples were cultivated from each cervid species in order to isolate the indicator bacterial species E. coli and Enterococcus faecalis/E. faecium for antibiotic resistance pattern studies. Salmonella and the potentially human pathogenic verocytotoxic E. coli were not isolated, while Campylobacter jejuni jejuni was found in one roe deer sample only. Antibiotic resistance was found in 13 (7.3%) of the 179 E. coli isolates tested, eight of these being resistant against one type of antibiotic only. The proportion of resistant E. coli isolates was higher in wild reindeer (24%) than in the other cervids (2.2%). E. faecalis or E. faecium were isolated from 19 of the samples, none of these being reindeer. All the strains isolated were resistant against one (84%) or more (16%) antibiotics. A total of 14 E. faecalis-strains were resistant to virginiamycin only. The results indicate that the cervid species studied do not constitute an important infectious reservoir for either the human pathogens or the antibiotic resistant microorganisms included in the study.     

乐果降解菌的分离、筛选和鉴定

筛选高效的有机磷农药乐果的降解菌。采集福建省福农生化有限公司排污口,沟口及草坪土壤进行乐果降解菌的分离鉴定。经含乐果的培养基分离培养、脂肪酸鉴定,总共得到17个属23株细菌,对这些菌通过消解试验进行复筛,得到4株对乐果有较好降解能力的菌株,沙门氏菌、阴沟肠杆菌、铜绿假单胞菌和枯草芽孢杆菌,在乐果浓度为1000mg/L时,48h的降解率分别为27.3%、26.6%、22.9%和18.1%。对乐果有降解作用的菌株存在种类多样性的特点。     

表达绿色荧光蛋白的重组减毒鼠伤寒沙门氏菌感染动力学

用表达绿色荧光蛋白(green fluorescent protein，GFP)的重组减毒鼠伤寒沙门氏菌X4550(pYAGFP)给BALB／c小鼠尾静脉注射和口服，时小鼠脏器进行细菌计数，并用间接ELISA法测定血清中抗细菌的抗体。结果表明，静脉注射重组菌的小鼠在肝脏、脾脏中重组菌数量有2个峰值，总的趋势是越来越少，而血清中抗体效价越来越高，可以初步推断，肝脏、脾脏中的细菌数量与血清中的抗体水平呈一定的相关性；口服重组菌的小鼠，在各脏器中重组菌的数量呈现正态分布的变化趋势，其中派伊尔结(Peyer’s patches，PPs)中细菌数量与血清中的抗体水平呈一定的相关性，而脾脏、肝脏及肠系膜淋巴结中细菌数量与血清中的抗体滴度无关。     

Nationwide surveillance of salmonella in the faeces of pigs in Japan

The prevalence of faecal carriage of salmonella in 5393 pigs reared on 218 pig farms located in 31 of 47 prefectures in Japan over the period July 2003 to June 2005 was investigated. We isolated 172 strains belonging to 20 serovars and one untypable Salmonella enterica from 169 pig faecal samples (3.1%) collected from 48 farms (22.0%). The most prevalent type of S. enterica was untypable O4,12:d:- which lacks phase 2 flagellar antigen, representing 29.1% (50/172) of all isolates. Of 26 S. enterica serovar Typhimurium isolates, 16 strains appeared to be definitive phage type 104 (DT104) by polymerase chain reaction.     

北京地区健康肉鸡携带沙门氏菌状况调查

禽源沙门氏菌是一种重要的人畜共患病原菌和食源性病原菌。肉鸡作为人肠炎沙门氏菌感染的主要来源,目前国内对其沙门氏菌携带状况的研究资料较少。为了解北京地区健康肉鸡中沙门氏菌的携带情况,本研究共采集该地区28个肉鸡养殖场的盲肠或泄殖腔拭子样品1310份,进行沙门氏菌的分离和鉴定,总共分离出沙门氏菌54株。血清分型结果表明,肠炎和爪哇安那为两种优势血清型,所占比例分别为31.5%和25.9%;其次是禽伤寒,占比9.3%。本研究为北京地区肉鸡场沙门氏菌病的防控及首都公共卫生安全提供了基础资料。     

基于CRISPR序列的肠道沙门氏菌分离株CSST分型

为验证一种新的CRISPR分型方法——CSST分型方法分型沙门氏菌的可行性,分别采用传统CRISPR与CSST两种分型方法,对20株临床肠道沙门氏菌分离株进行分型.结果显示:CSST分型方法同传统CRISPR分型方法分型沙门氏菌的结果完全一致,均将20株临床肠道沙门氏菌菌株分成了15种亚型,其中CR4/CT4和CR9/...