Yersinia enterocolitica. Isolation from faeces of adults and children in Queensland

Over a 22-month period, all 5075 faecal specimens received by the microbiology laboratory of the Mater Misericordiae Public Hospital, Brisbane, were screened for the presence of Yersinia enterocolitica by culturing on cefsulodinirgasan-novobiocin agar medium. Y. enterocolitica was isolated from 37 patients (0.7% of all specimens). Ten patients were 10 years of age or older, and presented predominantly with pain in the abdomen; of these, five underwent an appendicectomy. Mesenteric adenitis was diagnosed histologically in three patients and Y. enterocolitica was isolated from a lymph-node biopsy specimen in one of these. The other 27 patients were under 5 years of age, and presented predominantly with diarrhoea of 24 hours' to six weeks' duration. In seven cases, another recognized faecal pathogen was also isolated. The isolation rate of Y. enterocolitica was one-quarter that of Salmonella and seven times that of Shigella.     

小肠结肠炎耶尔森菌O:3血清型喹诺酮耐药的分子机制研究

目的 研究中国部分地区小肠结肠炎耶尔森菌菌株的耐药特征,为小肠结肠炎耶尔森菌感染的预防控制工作提供依据.方法 自2002年至2007年,选择几个省市在不同季节对腹泻人群进行了该病原菌的分离,并对分离到的菌株进行血清学分型、生物学分型、药敏试验以及gyrA和parC基因突变检测.结果 通过对该菌喹诺酮耐药决定区gyrA和parC基因的变异情况检测发现,在22株萘啶酸耐药的菌株中,有20株gyrA基因检出氨基酸变异,其中83位点共检出Ser(AGC)→Arg(AGA或AGG)突变12株,Ⅱe(ATC)突变3株,Cys(TGC)1株,87位点检出Gly(GGC)突变2株,Tyr(TAC)和Asn(AAC)突变各1株;3株萘啶酸敏感O:3血清型小肠结肠炎耳耶尔森菌未检出上述氨基酸位点突变,而parC基因无论耐药株或敏感株均未检出突变.结论 我国小肠结肠炎耶尔森菌对萘啶酸的耐药比较严重,这可能是因为氟喹讲酮类抗生素是临床最常用药物之一,而高水平用药,用药时间过长,以及动物饲料中抗生素的使用可能是小肠结肠炎耶尔森菌对喹诺酮类抗生素耐药的根本原因.氟喹诺酮的过度应用使萘啶酸耐药株选择性形成.     

Comparison of lipopolysaccharide and protein immunogens from pathogenic Yersinia enterocolitica bio-serotype 1B/O:8 and 2/O:9 using SDS-PAGE

Objective Yersinia enterocolitica is an extracellular pathogen and its related antigens interact with the host immune system.We investigated the difference in immunological characteristics between a highly pathogenic and poorly pathogenic strain of Y.enterocolitica.Methods We used SDS-PAGE and western blotting to characterize lipopolysaccharide (LPS),Yersinia outer membrane proteins (Yops),membrane proteins,and whole-cell proteins from poorly pathogenic Y.enterocolitica bio-serotype 2/O:9,isolated from China,and highly pathogenic bio-serotype 1B/O:8,isolated from Japan.Results These two strains of Y.enterocolitica had different LPS immune response patterns.Comparison of their Yops also showed differences that could have accounted for their differences in pathogenicity.The membrane and whole-cell proteins of both strains were similar; immunoblottting showed that the 35 kD and perhaps the 10 kD proteins were immunogens in both strains.Conclusion The major antigens of the two strains eliciting the host immune response were the LPS and membrane proteins,as shown by comparing protein samples with reference and purified preparations.     

丽江市家犬中致病性耶尔森菌流行病学调查

目的 了解丽江市鼠疫疫源地内家犬中携带致病性耶尔森菌的流行情况及其病原学特征。方法 选择丽江市古城区和玉龙县各2个乡镇,逐户采集犬肛拭子及血液标本。肛拭子经4 ℃冷增菌后,采用耶尔森选择培养基对小肠结肠炎耶尔森菌、鼠疫耶尔森菌和假结核耶尔森菌三种致病性耶尔森菌进行分离;对所分菌株再进行生化鉴定、生物分型、血清分型和毒力基因型检测;对血液标本进行鼠疫F1抗体的检测。结果 共采集467份肛拭子,分离到4株小肠结肠炎耶尔森菌,未分离到鼠疫耶尔森菌及假结核耶尔森菌;4株小肠结肠炎耶尔森菌中,2株为O∶3血清型和非1A生物型,毒力基因齐全为致病株;另2株为1A生物型血清型未定,毒力基因阴性为非致病株;467份血液标本中26份为F1抗体阳性,其中1份阳性标本与1份分离到小肠结肠炎耶尔森菌的肛拭子来自同一只家犬。结论 丽江市家犬中携带小肠结肠炎耶尔森菌,且有致病性菌株;此外,在一只家犬中,发现存在着鼠疫耶尔森菌与小肠结肠炎耶尔森菌共感染的现象。     

Serum anti-Yersinia antibody in Chinese patients with Kawasaki disease

BACKGROUND: Many infectious agents have been implicated as an etiology to develop Kawasaki disease (KD). In Taiwan, studies on the relationship between Yersinia and KD have not been reported. METHODS: We measured sera for anti-Yersinia antibodies by using enzyme immunoassay (EIA) in 31 patients with KD and 60 healthy children (HC). Yersinia strains included Y. pseudotuberculosis I, II, III, IV, V, VI and Y. enterocolitica O3, O8 and O9. RESULTS: Data of 31 patients with KD showed that for the IgG antibody, serum anti-Y. pseudotuberculosis II, III, Y. O8 and O9 antibody were significantly higher when compared to the HC. Except for Y. pseudotuberculosis IV, all other Yersinia strains of either IgA or IgM antibodies increased significantly in patients with KD vs. the HC. If we compared the number of patients who had significant elevation of OD and those of HC, we found IgA anti-Yersinia antibodies (PST I, PST II, O3, O8, O9), IgM (PST VI, O8) and IgG (PST II, O8, O9) were significantly elevated in KD patients than in HC. A significant relationship was present between KD with myocarditis and increased anti-Yersinia antibody titer. CONCLUSIONS: The findings in this study suggest that preceding Yersinia infection may play a role in the pathogenesis of KD. Further study of the relationship between KD with myocarditis and increased anti-Yersinia antibody is needed.     

产毒性和致病性大肠杆菌中小肠结肠炎耶尔森菌强毒力岛的检测

目的　了解小肠结肠炎耶尔森菌强毒力岛在产毒性和致病性大肠杆菌中的分布,探讨毒力岛在大肠杆菌中的结构和功能。方法　对毒力岛的关键基因irp2、fyua和asn-intB进行PCR扩增和DNA测序,并对irp2和fyua进行原位杂交。结果　在检测的93株肠产毒性大肠杆菌和10株肠致病性大肠杆菌基因irp2的检出率分别为32.25％（30/93）和30％（3/10）,fyua的阳性率为21.51％（20/93）和30％（3/10）,而且这些阳性菌株中的毒力岛大部分连接到天门冬氨酸tRNA（asn tRNA）位点。结论　小肠结肠炎耶尔森菌强毒力岛在产毒性和致病性大肠杆菌中较高的阳性率,对于大肠杆菌毒力的变化和毒力的调控以及细菌毒力的进化可能具有重要意义。     

国内首次发现携带耶尔森菌HPI毒力岛rip^-2也基因的阴沟肠杆菌

目的研究感染性腹泻患者粪便中分离的阴沟肠杆菌携带的小肠结肠炎耶尔森菌HPI毒力岛irp-^2基因，并评价其菌株的毒力和耐药情况。方法PCR扩增法检测毒力岛irp^-2基因，小白鼠腹腔注射法捡测毒力，药敏试验采用K—B纸片扩散法。结果从感染性腹泻患者粪便中分离的9株阴沟肠杆菌均扩增出irp^-2基因片段，其相对分子量与对照菌株小肠结肠炎耶尔森菌WA株（08型）的相应PCR片段一致，并具有较强的毒力。结论检出的阴沟肠杆菌含有小肠结肠炎耶尔森菌HPI毒力岛irp^-2基因且为具有毒力的菌株，与致病性密切相关。     

产毒性和致病性大肠杆菌中小肠结肠炎耶尔森菌强毒力岛的检测

目的 了解小肠结肠炎耶尔森菌强毒力岛在产毒性和致病性大肠杆菌中的分布，探讨毒力岛在大肠杆菌中的结构和功能。方法 对毒力岛的关键基因irp2,fyua和asn-intB进行PCR扩增和DNA测序，并对irp2和fyua进行原位杂交。结果 在检测的93株肠产毒性大肠杆菌的10株肠致病性大肠杆菌基因irp2的检出率分别为32.25%(30/93)和30%（3/10),fyua的阳性率为21.51%(20/93)和30%（3/10）。而且这些阳性菌株中的毒力岛大部分连接到天门冬氨酸tRNA(asntRNA）位点。结论 小肠结肠炎耶尔森菌强毒力岛在产毒性和致病性大肠肝菌中较高的阳性率，对于大肠杆菌毒力的变化和毒力的调控以及细菌毒力的进化可能具有重要意义。     

Bacterial enteropathogens isolated in childhood diarrhoea in Kuala Lumpur--the changing trend

A retrospective review of all stool samples obtained from children aged < 16 years with diarrhoea from University of Malaya Medical Centre (UMMC), Kuala Lumpur, from 1978 to 1997 was undertaken to ascertain the pattern of bacterial pathogens causing diarrhoea in children in an urban area in Malaysia. Of 26444 stool samples processed, 2989 (11%) were positive. The five most common bacterial pathogens isolated were non-typhoidal Salmonella (57%), enteropathogenic E. coli (EPEC) (14%), Shigella spp. (11%), Campylobacter spp. (5%) and Aeromonas spp. (4%). There was a significant reduction in the average percentage of positive isolation during the last 5 years of the study period as compared to the first 5 years (15.0% vs. 7.2%; r = -0.92, p = 0.0001). EPEC and Shigella spp. were less commonly isolated in the last five years compared with the first five years of the study (6% vs 21% p < 0.001 for E. coli; 7% vs 22%, p < 0.001 for Shigella spp.). This information is important for public health education in reducing the incidence of childhood diarrhoea further, and in the selection of appropriate antimicrobials in the management of extra-intestinal complications of childhood diarrhoea.     

Cryptosporidiosis and childhood diarrhoea in a rural community in Kenya

Cryptosporidiosis has come to recent prominence as a cause of diarrhoea throughout the world in both adults and children, producing either an acute self-limiting diarrhoea or a protracted chronic diarrhoea which can be fatal in immunosuppressed patients. This study was therefore conducted to assess the prevalence of the infection among children less than five years of age in a rural community in Kiambu District, Kenya. 1420 diarrhoea specimens from children less than five years which were processed for bacteriology and parasitology were also examined for Cryptosporidium oocysts, using the modified Ziehl-Neelsen (ZN) acid-fast stain, 3.8% of all the diarrhoea samples were positive for oocysts. 320 non-diarrhoeic stools from children in the same age group were also examined and were all negative for Cryptosporidium oocysts. The results of this study would imply that infection with Cryptosporidium is associated with acute childhood diarrhoea in Kenya.     

A multistate outbreak of infections caused by Yersinia enterocolitica transmitted by pasteurized milk

In June and July 1982, a large interstate outbreak of Yersinia enterocolitica infections caused by an unusual serotype occurred in Tennessee, Arkansas, and Mississippi. Eighty-six percent of cases had enteritis characterized by fever, abdominal pain, and diarrhea. In three separate case-control studies, drinking milk pasteurized by plant A was statistically associated with illness. In a survey of randomly chosen households, 8.3% of persons who recalled having drunk milk from plant A during the suspect period experienced a yersiniosislike illness. Inspection of the plant and cultures of the available raw and pasteurized milk did not reveal the source or mechanism of contamination or a breach in normal pasteurizing technique. Although outbreaks of enteric disease caused by pasteurized milk are rare in the United States, the ability of Y enterocolitica to grow in milk at refrigeration temperatures makes pasteurized milk a possible vehicle for virulent Y enterocolitica. The extent to which milk is responsible for sporadic cases of yersiniosis is unknown.     

Pleiotropic effect of tatC mutation on metabolism of pathogen Yersinia enterocolitica

Objective To analyze the impact of depletion of the twin arginine translocation （TAT） system on virulence and physiology of Yersinia enterocolitica for a better understanding of its pathogenicity. Methods We constructed a △tatC：：Sp^R mutant of Yersinia enterocolitica by P1 phage mediated transduction using Escherichia coli K-12 △tatC：：Sp^R strain as a donor. Results A Pl-mediated genetic material transfer was found between the two species of enterobacteria, indicating a great potential of acquisition of antibiotic resistance in emergency of a new threatening pathogen by genetic material exchanges. Periplasmic trimethylamine N-oxidase reductase activity was detected in the wild type E enterocolitica strain and translocation of this enzyme was completely abolished by the △tatC：：Sp^R mutation. In addition, the △tatC：：Sp^R mutation showed a pleiotropic effect on the metabolism of E enterocolitica. However, the tat mutation did not seem to affect the mobility and virulence of Y. enterocolitica under the conditions used. Conclusion Unlike other pathogenic bacteria studied, the TAT system of E enterocolitica might play an important role in the pathogenic process, which is distinct from other pathogens, such as Pseudomonas aeruginosa and enterohemorrhagic E. coli O 157：H7.     

Yersinia enterocolitica enteritis in children.

Five patients were admitted to Children's Hospital National Medical Center with manifestations of Yersinia enterocolitica infections. Four of five had primarily enteric illness, while the fifth had associated ocular and joint involvements. In three of five cases, the disease was self-limiting. Two of the patients (No. 1 and 2) may have had their disease improved by antibiotic therapy. Increased alertness to the potential of Y enterocolitica to cause a variety of syndromes is necessary to make an accurate bacteriologic diagnosis and to expedite medical treatment.     

建立环介导等温扩增方法检测腹泻患者中小肠结肠炎耶尔森菌

目的建立快速的检测方法对腹泻患者中的小肠结肠炎耶尔森菌进行快速检测。方法利用析因设计试验方法建立并优化检测小肠结肠炎耶尔森菌的环介导等温扩增技术（LAMP）,并与普通PCR法对临床腹泻标本进行检测比较。结果析因设计试验确定LAMP最佳反应体系：Mg2＋＋为2．0mmol,内引物FIP／BIP为1．6umol,dNTPs为1．6mmol,时间为60rain,反应温度为63℃。对12种细菌进行LAMP扩增,仅小肠结肠炎耶尔森菌获得阳性扩增结果,小肠结肠炎耶尔森菌基因组DNA和纯培养物的检测灵敏度分别为38．05fg和15CFU／ml。对腹泻样本进行直接检测,检测限为150CFU／g。539例腹泻患者粪便标本中,LAMP成功检出13例样本中含有小肠结肠炎耶尔森菌,普通PCR检出11例,灵敏度为100％,特异度为99．62％。结论本方法检测小肠结肠炎耶尔森菌特异性强、灵敏度高．且操作简单、快速、检测成本低．适合基层单位及现场廊用．     

Species distribution and antibiotic resistance of shigella isolates in an urban community in Malaysia

There is an increasing trend for Shigella isolates worldwide to be resistant to commonly prescribed antibiotics. The species distribution and antibiotic resistance of Shigella species isolated from children in Kuala Lumpur, Malaysia from 1978 to 1997 was reviewed. Three hundred and eighty six isolates were positive for Shigella species, representing 1.4% (95% CI: 1.3%-1.6%) of the 26320 total stool specimens and 13% (95% CI: 11.8%-14.2%) of 2986 isolates positive for bacterial pathogens. Shigella flexneri, constituting 74% of all isolates in the first five years of the study, decreased by 40% during the last five years (95% CI of decrease: 22.1%-57.9%), p-value < 0.0001) to 34%. There was a significant reduction (chi2 for linear trend = 77.6, p-value < 0.001) in the number of Shigella isolates as a percentage of total stool isolates obtained. 58% of the 241 isolates tested for antibiotic sensitivity were resistant to at least one antibiotic, and 42% wEre multi-resistant to three or more antibiotics. Shigella species was not a common pathogen among children admitted with diarrhoea in Kuala Lumpur, and was more likely to be resistant to commonly prescribed antibiotics.     

食源致病菌96 微孔板DNA 诊断芯片的研制及在一起食物中毒突发事件中的应用

目的研制能同时检测9种常见食源致病菌的96微孔板DNA诊断芯片。方法针对金黄色葡萄球菌、沙门氏菌、大肠杆菌O157:H7(Stx1和Stx2)、志贺氏菌、产单核细胞李斯特菌、蜡样芽胞杆菌、小肠结肠炎耶尔森菌、副溶血弧菌、霍乱弧菌等9种最常见的食源性致病菌,首先选择合适的保守基因,设计种特异性的PCR引物(5'端标记生物素)和检测探针,通过参数优化建立一管多重PCR扩增体系;然后按5×5的阵列格式将探针点制到96微孔板的微孔内,通过条件优化建立稳定的PCR产物与固化探针的微孔杂交体系;采用链霉抗生物素蛋白碱性磷酸酶和化学显色底物NBT/BCIP来检测特异性的PCR杂交产物。结果20株标准菌株验证已建立的食源致病菌微孔板DNA诊断芯片,获得比较特异和稳定的实验结果。在一起食物中毒事件中,该芯片在采样后12h内检出金黄色葡萄球菌,与传统的细菌分离培养和生化鉴定的结果相符。结论本研究建立的新型食源致病菌96微孔板DNA诊断芯片具有快速、准确、自动化和高通量等特点,为快速应对食物中毒等突发公共卫生事件提供了非常有价值的检测技术。     

Prevalence of bacterial pathogens and susceptibility patterns from clinical sources in Trinidad

During a 12-month period (January-December, 1997), bacterial isolates of specimens from in-patients and out-patients of the Eric Williams Medical Sciences Complex (EWMSC) were reviewed. A total of 3,513 specimens were processed, 43.1% from in-patients and 56.9% from out-patients. Of the 3,513 specimens, 1129 (32.1%) yielded positive cultures. Micro-organisms from wounds, sputum and genital tract accounted for 90.2%, 51.5% and 31.8%, respectively, of all isolates. E coli (17.4%) and Enterococci (12.2%) were the predominant isolates and were also the major pathogens from blood stream infections, 25.8% and 18.2%, respectively, followed by P aeruginosa, 15.2%. High levels of resistance were seen to ampicillin, augmentin (amoxicillin-clavulanic acid) and tetracycline. The most effective antibiotics were ceftazidime (no resistance in E coli Citrobacter spp, non-typhoidal Salmonella and Group B streptococci, 63.2% resistance in Acinetobacter spp, 15.2% in Enterobacter spp, 17.4% in Klebsiella spp.], gentamicin [no resistance in Enterobacter and Citrobacter spp, and 89.5% in Acinetobacter spp), erythromycin (no resistance in Groups A and B streptococci, 85.1% in S aureus and S pneumoniae). The spectrum of isolates will provide clinicians with data on which to base their "best guess" aetiologic agent and choice of antibiotics when faced with infectious diseases in areas where laboratory assistance is not readily available.     

多重聚合酶链反应检测食品中致病性小肠结肠炎耶尔森菌和伤寒沙门氏菌

[目的 ]快速准确地检测致病性小肠结肠炎耶尔森菌和伤寒沙门氏菌 .[方法 ]根据小肠结肠炎耶尔森菌的ail基因和ST的H1 d基因顺序设计二对引物 ,通过聚合酶链反应检测食品中的小肠结肠炎耶尔森菌和伤寒沙门氏菌 ,扩增产物经电泳 ,分别出现与已知阳性对照细菌条带大小一致的 170和 4 5 8bp的ail和H1 d基因片断的条带时即判定该样品为阳性 .[结果和结论 ]本法特异性强、灵敏度高 ,能在60h内出结果 ,适用于食品中小肠结肠炎耶尔森菌和伤寒沙门氏菌的快速检验 .     

Mixed infections in childhood diarrhoea: results of a community study in Kiambu District, Kenya

One thousand four hundred and twenty diarrhoea specimens from 846 children aged 0 to 60 months were collected and analysed for bacteria, parasites and rotavirus over a 16 month period, from June 1985 to September 1986 inclusive. The study was conducted in 4 villages situated in Kiambu District, Kenya. All the specimens were analysed for rotavirus and parasites, including Cryptosporidium. The majority of the specimens were analysed for enteropathogenic Escherichia coli (EPEC), Shigella, Salmonella, Campylobacter and Aeromonas. Only 387 specimens were analysed for enterotoxigenic E. coli (ETEC). However, of this proportion analysed for ETEC, 33% were positive. A total of 344 specimens were negative for any organisms while a further 140 were only positive for parasites which have been implicated as being pathogenic, including Ascaris lumbricoides, Hymenolepis nana, Trichomonas hominis and Blastocysts hominis were considered to be at least potentially pathogenic and capable of causing diarrhoea. An average of only 29.4% of these organisms occurred as single isolates. The remaining infections were mixed, with a maximum of 7 potentially pathogenic organisms occurring together in a single specimen. The associations of certain organisms were significant, notably Campylobacter with Giardia lamblia. Campylobacter with EPEC, EPEC with Ascaris, and G. lamblia with rotavirus. The latter was a negative association.     

实时定量聚合酶链反应检测腹泻粪便中小肠结肠炎耶尔森菌的研究与评价

目的 建立一种快速准确的分子生物学方法检测腹泻粪便中小肠结肠炎耶尔森菌,以替代繁琐费时的培养鉴定方法.方法 应用针对致病性小肠结肠炎耶尔森菌染色体上yst基因而设计的特异引物和探针进行实时定量聚合酶链反应（PCR,探针法）,检测致病性（7种血清型）和非致病性（5种）小肠结肠炎耶尔森菌、耶尔森菌属内的其他菌种（8种）及其他肠道细菌（8种）.此外,实时定量PCR检测经过培养方法鉴定的腹泻粪便200例,并比较两种方法的吻合度.结果 引物和探针对致病性小肠结肠炎耶尔森菌100%特异,与其他细菌无交叉反应.在纯培养物和粪便中,最低检测浓度分别为102菌落形成单位（CFU）/ml和103 CFU/g.在200例样本中,培养鉴定方法和实时定量PCR检出同样18例阳性样本,吻合度为100%.结论 实时定量PCR反应对致病性小肠结肠炎耶尔森菌高度特异且与培养鉴定方法有很好的吻合性又方便快捷,可应用于临床检测.