Vibrio cholerae O139: How great is the threat of a pandemic?

The emergence of the new strain Vibrio cholerae O139 and its rapid spread in Bangladesh and India together with its detection in several other countries, have raised the question whether this constitutes the beginning of the eighth pandemic of cholera, and if so, how large a threat it poses. In an attempt to answer this question, epidemic spread patterns of Vibrio cholerae O139 strain in Bangladesh were studied. Initially the epidemic moved quickly and affected the entire coastal and estuarine tidal plains of southern Bangladesh. In the flood plains of the northern regions it affected mostly the north‐eastern and north‐central areas, at a slower pace than in the southern areas. In the beginning the new strain totally displaced both biotypes (classic and El Tor ) of Vibrio cholerae O1. Nearly 2 years after its initial detection, striking differences in the distribution of V. cholerae O139 and O1 were observed. In most northern areas, the new strain was replaced by V. cholerae O1, whereas in the southern coastal regions, the O139 strain continues to dominate epidemics. The study suggests that the O139 strain may become endemic in the coastal ecosystem. The threat of a pandemic, therefore, may not be as large as it first seemed.     

Emergence of tetracycline-resistant Vibrio cholerae O1 serotype Inaba, in Kolkata, India

Out of 2,235 diarrheal stool samples collected from patients admitted to the Infectious Diseases Hospital, Kolkata, 343 cases were positive for Vibrio cholerae (341, V. cholerae O1 and 2, O139). During the year 2004, infections caused by V. cholerae serotype Ogawa and Inaba were 93 and 7%, respectively, while in 2005, the Inaba isolation rate rose to 88% as compared to 12% for Ogawa. Susceptibility to antimicrobial agents revealed that the O1 strains were resistant to multiple antibiotics (ampicillin, co-trimoxazole, furazolidone, nalidixic acid and streptomycin) with reduced susceptibility to ciprofloxacin. Increased isolation of tetracycline-resistant strains (27.3% for Ogawa and 15% for Inaba) was noted in 2005. It appears that the population might be at risk of infection by the Inaba serotype and that tetracycline may not be useful for the treatment.     

Increasing drug resistance in Vibrio cholerae O1 and non-O1 strains isolated from diarrheal cases in Japan

Drug resistance trends were investigated for 271 Vibrio cholerae O1 (V.c O1) and 401 V. cholerae non-O1 (V.c non-O1) strains isolated from mainly imported diarrheal cases during 1981-2001 in Japan. The results of drug resistance test using 8 drugs (CP, TC, SM, KM, ABPC, ST, NA, and NFLX) showed that 34.7% of the V. c O1 strains and 15.7% of V.c non-O1 strains were multi-drug or mono-drug resistant. The incidence of drug resistant strains has increased since 1991, and it has been remarkable in V.c O1 strains that increased from 1.2% in 1981-1985 to 70.8% in 1996-2001. The drug resistance patterns of the resistant strains classified into 6 types in V.c O1 and 21 types in V.c non-O1. The prevalent patterns recognized were SM (75.5%), CP.TC.SM.ST (10.6%) and CP.SM.ST (8.5%) in V.c O1, and SM (25.4%) and ABPC (25.4%) in V.c non-O1. Ten V.c O1 strains (3.7%) and 10 V.c non-O1 strains (2.5%) were multi-drug resistant including TC. Among those, 13 strains were isolated from travelers who returned to Japan from Thailand. One V.c O1 strain (0.4%) and 6 V.c non-O1 strains (1.5%) were NA high-resistant and fluoroquinolones low-sensitive. Among those, 4 strains were isolated from travelers who returned to Japan from India.     

霍乱弧菌O1群和O139群nhaA基因的克隆和变异性研究

目的 克隆霍乱弧菌O1群和O139群nhaA基因，并分析其变异性。方法收集我国1988—2000年散发霍乱弧菌O1群和O139群40株，用聚合酶链反应(PCR)扩增nhaA基因，克隆至pcDNA3载体，通过序列测定分析其同源性和变异性。结果 分别从霍乱弧菌Ol群和O139群扩增出约1．2kb的nhaA基因片段，基因序列分析表明，我国霍乱散发O1群和O139群的nhaA基因与Genbank中霍乱弧菌O1群野毒株参考序列同源性分别为99％和96％。编码氨基酸的突变率分别为2％和11％，nhaA基因重要的结构和功能决定簇(Aspl33、Aspl63、Aspl64、His225、Leu73和Gly338)未发生变异；第203、221位的氨基酸，O1群和O139群发生相同的变异。结论 nhaA基因编码氨基酸的变异可能是霍乱弧菌适应外环境变化的结果。     

Shifting prevalence of major diarrheal pathogens in patients seeking hospital care during floods in 1998, 2004, and 2007 in Dhaka, Bangladesh

Bangladesh experienced severe flooding and diarrheal epidemics in 2007. We compared flood data from 2007 with 2004 and 1998 for diarrheal patients attending the ICDDR,B hospital in Dhaka. In 2007, Vibrio cholerae O1 (33%), rotavirus (12%), and enterotoxigenic Escherichia coli (ETEC) (12%) were most prevalent. More severe dehydration was seen in 2007 compared with 2004 and 1998 (P < 0.001). In 2007, V. cholerae O1 Inaba (52%) and Ogawa (48%) were seen, whereas in 2004 and 1998 it was primarily Inaba and the Ogawa types, respectively (P < 0.001). In 2007, 51% of ETEC produced the heat labile toxin (LT) (P < 0.001 compared with 2004), 22% expressed the heat stable (ST) (P < 0.001), and 27% were ST/LT positive (P = 0.231). The CS7 colonization factor (CF) was the most prevalent in 2007 (20% compared with 6% in 2004; P = 0.05). Our findings demonstrate alterations in clinical features and phenotypic changes of major bacterial pathogens in the recent Bangladesh flood.     

Antimicrobial-resistant bacterial diarrhea in rural western Kenya

Bacterial diarrheal diseases cause substantial morbidity and mortality in sub-Saharan Africa, but data on the epidemiology and antimicrobial susceptibility patterns of enteric bacterial pathogens are limited. Between May 1997 and April 1998, a clinic-based surveillance for diarrheal disease was conducted in Asembo, a rural area in western Kenya. In total, 729 diarrheal specimens were collected, and 244 (33%) yielded >or=1 bacterial pathogen, as determined by standard culture techniques; 107 (44%) Shigella isolates, 73 (30%) Campylobacter isolates, 45 (18%) Vibrio cholerae O1 isolates, and 33 (14%) Salmonella isolates were identified. Shigella dysenteriae type 1 accounted for 22 (21%) of the Shigella isolates. Among 112 patients empirically treated with an antimicrobial agent and whose stool specimens yielded isolates on which resistance testing was done, 57 (51%) had isolates that were not susceptible to their antimicrobial treatment. Empiric treatment strategies for diarrheal disease in western Kenya need to be reevaluated, to improve clinical care.     

Emergence and evolution of Vibrio cholerae O139

The emergence of Vibrio cholerae O139 Bengal during 1992-1993 was associated with large epidemics of cholera in India and Bangladesh and, initially, with a total displacement of the existing V. cholerae O1 strains. However, the O1 strains reemerged in 1994 and initiated a series of disappearance and reemergence of either of the two serogroups that was associated with temporal genetic and phenotypic changes sustained by the strains. Since the initial emergence of the O139 vibrios, new variants of the pathogen derived from multiple progenitors have been isolated and characterized. The clinical and epidemiological characteristics of these strains have been studied. Rapid genetic reassortment in O139 strains appears to be a response to the changing epidemiology of V. cholerae O1 and also a strategy for persistence in competition with strains of the O1 serogroup. The emergence of V. cholerae O139 has provided a unique opportunity to witness genetic changes in V. cholerae that may be associated with displacement of an existing serogroup by a newly emerging one and, thus, provide new insights into the epidemiology of cholera. The genetic changes and natural selection involving both environmental and host factors are likely to influence profoundly the genetics, epidemiology, and evolution of toxigenic V. cholerae, not only in the Ganges Delta region of India and Bangladesh, but also in other areas of endemic and epidemic cholera.     

O139霍乱弧菌肠毒素核苷酸序列分析

目的 探讨Ｏ１３９霍乱弧菌肠毒素（ＣＴＸ）核苷酸与Ｏ１群霍乱ＣＴＸ毒素核苷酸序列异同。方法 用聚合酶链反应、ＤＮＡ序列分析测定２株Ｏ１３９群、２株Ｏ１群古典型、２株Ｏ１群Ｅ１Ｔｏｒ型霍乱弧菌ＣＴＸＡ２－Ｂ亚单位核苷酸。结果 ２株Ｏ１３９群霍乱弧菌均含有ＣＴＸＡ２－Ｂ亚单位基因,Ｏ１３９群与Ｏ１群ＣＴＸＡ２－Ｂ核苷酸同源性为９７．１％～９８．９％结论 Ｏ１３９群与Ｏ１群霍乱弧菌ＣＴＸ核苷酸序列一致。     

河南省某一淡水养殖环节中非O1/O139群霍乱弧菌毒力基因及分子分型分析

目的 了解河南省淡水养殖环节中非O1/O139群霍乱弧菌毒力基因分布及分子分型情况。方法 对河南省淡水养殖环节中50株非O1/O139群霍乱弧菌和3株病人来源菌株进行全基因测序,利用PubMLST-Vc数据库分析其序列分型(ST),利用最小生成树关系图分析进化关系,通过VFDB数据库获得其毒力基因分布。结果 来源于淡水养殖环节和来源于病人的非O1/O139群霍乱弧菌53株菌株均具有黏附、趋化运动、抗吞噬、毒素及酶类等功能的8类毒力相关因子基因,缺失辅助定植、毒素共调、分泌等功能的毒力相关因子基因和副霍乱肠毒素等4个毒素基因。部分毒力相关因子或部分菌株的毒力相关因子毒力基因不全。与3株来源于病人的菌株相比,二者毒力因子基因携带情况相近,除MSHA Ⅳ型菌毛毒力因子mshA基因、荚膜多糖wbuB基因、wbfY基因、rmlB基因、血红素受体hutA基因及Ⅲ型分泌系统vscC2和vcrD2基因外,部分菌株二者携带相同的毒力因子基因。53株非O1/O139群霍乱弧菌分属19个ST型,ST4和ST5是优势ST型,养殖环节来源的菌株与病人来源的菌株分属不同的ST型。19个ST型等位基因位点变异差异数在1~7个,其中分离自淡水养殖环节的非O1/O139群霍乱弧菌菌株17个ST型等位基因位点变异差异数在1~7个,病人来源的非O1/O139群霍乱弧菌菌株2个ST型与分离自淡水养殖环节的非O1/O139群霍乱弧菌菌株ST1、ST2、ST6和ST10属同一簇,与ST1有6个等位基因位点存在差异。结论 河南省淡水养殖环节非O1/O139群霍乱弧菌菌株MLST分型多样化,携带多种毒力相关因子,不同来源菌株携带的毒力基因相同,虽然分属不同的ST型,但还是存在食品安全风险,提醒有关部门采取措施进行防控。     

2011—2018年厦门市集美区霍乱监测结果分析

目的 对厦门市集美区2011—2018年霍乱监测结果进行分析,了解集美区水产品和外环境水体中霍乱弧菌的污染状况,尽早发现霍乱病例,为采取有效的防治措施提供依据.方法 采集水产品、外环境水体及腹泻病人粪便标本,根据《全国霍乱监测方案》《霍乱诊断标准》(WS 289-2008)和《霍乱防治手册(第6版)》的规定进行检测分析...     

Are the environmental niches of Vibrio cholerae O139 different from those of Vibrio cholerae O1 El Tor?

BACKGROUND: Vibrio cholerae are known to be normal inhabitants of surface water. However, the environmental niches of the different strains of cholera are not well known, and therefore, populations at risk for cholera outbreaks cannot be clearly identified. METHODS: This study identifies environmental risk factors for cholera caused by V. cholerae O1 El Tor and O139 and environmental niches of the two strains present in Matlab, a cholera endemic area of Bangladesh. The study year was 1993, the year that the O139 strain first appeared in the study area. Patients who had either strain of cholera identified in a laboratory were included in the study. A geographic information system was used to map the household locations of the patients, to describe the human sanitary environment and population density, and to address potential anthropogenic and environmental risk factors of the disease. Spatial point pattern and exploratory spatial data analysis techniques were used to define the environmental niches of the two cholera strains. RESULTS: The study suggests the niches of O1 El Tor and O139 strains of V. cholerae appear to be similar, based on common environmental risk factors. CONCLUSIONS: The results of this study support a theory that O1 El Tor could possibly be replaced by the newer O139 strain in the future.     

Seasonal epidemics of cholera inversely correlate with the prevalence of environmental cholera phages

The relationship among (i) the local incidence of cholera, (ii) the prevalence in the aquatic environment of Vibrio cholerae, and (iii) bacterial viruses that attack potentially virulent O1 and O139 serogroup strains of this organism (cholera phages) was studied in Dhaka, Bangladesh. Over nearly a 3-year period, we found that significantly more environmental water samples contained either a phage or a phage-susceptible V. cholerae strain than both (P < 0.00001). The number of cholera patients varied seasonally during this period and frequently coincided with the presence of pathogenic V. cholerae strains in water samples that otherwise lacked detectable cholera phages. Interepidemic periods were characterized by water samples containing cholera phages but no viable bacteria. Our data support the conclusion that cholera phages can influence cholera seasonality and may also play a role in emergence of new V. cholerae pandemic serogroups or clones.     

Genomic diversity of 2010 Haitian cholera outbreak strains

The millions of deaths from cholera during the past 200 y, coupled with the morbidity and mortality of cholera in Haiti since October 2010, are grim reminders that Vibrio cholerae, the etiologic agent of cholera, remains a scourge. We report the isolation of both V. cholerae O1 and non-O1/O139 early in the Haiti cholera epidemic from samples collected from victims in 18 towns across eight Arrondissements of Haiti. The results showed two distinct populations of V. cholerae coexisted in Haiti early in the epidemic. As non-O1/O139 V. cholerae was the sole pathogen isolated from 21% of the clinical specimens, its role in this epidemic, either alone or in concert with V. cholerae O1, cannot be dismissed. A genomic approach was used to examine similarities and differences among the Haitian V. cholerae O1 and V. cholerae non-O1/O139 strains. A total of 47 V. cholerae O1 and 29 V. cholerae non-O1/O139 isolates from patients and the environment were sequenced. Comparative genome analyses of the 76 genomes and eight reference strains of V. cholerae isolated in concurrent epidemics outside Haiti and 27 V. cholerae genomes available in the public database demonstrated substantial diversity of V. cholerae and ongoing flux within its genome.     

Plasmid-borne multiple drug resistance in Vibrio cholerae serogroup O1, biotype El Tor: evidence for a point-source outbreak in Bangladesh

In 1979, an outbreak of plasmid-borne, multiply drug-resistant Vibrio cholerae serogroup O1 biotype El Tor (V. cholerae O1) occurred in the Matlab area of Bangladesh. The outbreak could have resulted from the introduction into the area of a single resistant strain or from multiple conjugations of drug-sensitive V. cholerae O1 with C plasmids in other environmental flora. Resistant strains were phage typed to determine their relatedness, and plasmid studies were conducted to determine the frequency of C plasmids in nonvibrio flora of family contacts of cholera patients. Forty-one (85%) of 48 resistant strains of V. cholerae O1 examined belonged to two closely related phage types new to the area, whereas 59 drug-sensitive strains from the same period were primarily of two different phage types. Group C plasmids were in nonvibrio strains from five of 36 family contacts of patients with drug-resistant cholera but none of 82 family contacts of patients with sensitive cholera. This outbreak most likely began from the introduction into the area of a single, multiply drug-resistant strain of V. cholerae O1. C plasmids detected in the nonvibrio flora of family contacts probably came from the resistant strain of V. cholerae O1.     

Genetic diversity and virulence potential of environmental Vibrio cholerae population in a cholera-endemic area

To understand the evolutionary events and possible selection mechanisms involved in the emergence of pathogenic Vibrio cholerae, we analyzed diverse strains of V. cholerae isolated from environmental waters in Bangladesh by direct enrichment in the intestines of adult rabbits and by conventional laboratory culture. Strains isolated by conventional culture were mostly (99.2%) negative for the major virulence gene clusters encoding toxin-coregulated pilus (TCP) and cholera toxin (CT) and were nonpathogenic in animal models. In contrast, all strains selected in rabbits were competent for colonizing infant mice, and 56.8% of these strains carried genes encoding TCP alone or both TCP and CT. Ribotypes of toxigenic O1 and O139 strains from the environment were similar to pandemic strains, whereas ribotypes of non-O1 non-O139 strains and TCP(-) nontoxigenic O1 strains diverged widely from the seventh pandemic O1 and the O139 strains. Results of this study suggest that (i) the environmental V. cholerae population in a cholera-endemic area is highly heterogeneous, (ii) selection in the mammalian intestine can cause enrichment of environmental strains with virulence potential, (iii) pathogenicity of V. cholerae involves more virulence genes than currently appreciated, and (iv) most environmental V. cholerae strains are unlikely to attain a pandemic potential by acquisition of TCP and CT genes alone. Because most of the recorded cholera pandemics originated in the Ganges Delta region, this ecological setting presumably favors extensive genetic exchange among V. cholerae strains and thus promotes the rare, multiple-gene transfer events needed to assemble the critical combination of genes required for pandemic spread.     

A 4-year study of the epidemiology of Vibrio cholerae in four rural areas of Bangladesh

How Vibrio cholerae spreads around the world and what determines its seasonal peaks in endemic areas are not known. These features of cholera have been hypothesized to be primarily the result of environmental factors associated with aquatic habitats that can now be identified. Since 1997, fortnightly surveillance in 4 widely separated geographic locations in Bangladesh has been performed to identify patients with cholera and to collect environmental data. A total of 5670 patients (53% <5 years of age) have been studied; 14.3% had cholera (10.4% due to V. cholerae O1 El Tor, 3.8% due to O139). Both serogroups were found in all locations; outbreaks were seasonal and often occurred simultaneously. Water-use patterns showed that bathing and washing clothes in tube-well water was significantly protective in two of the sites. These data will be correlated with environmental factors, to develop a model for prediction of cholera outbreaks.     

Enterotoxigenic Escherichia coli causing cholerogenic syndrome during an interepidemic period of cholera in North India

A study was undertaken to look for heat-labile enterotoxin producing Escherichia coli (LT-ETEC) in patients with acute watery diarrhea resembling cholera in Chandigarh in North India. Two periods were selected, an interepidemic (2001) and an epidemic (2002) period. Heat-labile enterotoxin (LT) was detected using reverse passive latex agglutination test. LT-ETEC strains were serotyped and antimicrobial susceptibility was studied. LT-ETEC was detected in 16 of 21 stool samples during 2001.Other organisms isolated included Vibrio cholerae O1 Ogawa and Aeromonas hydrophila in 2 and 3 patients, respectively. During the epidemic period, 4 of 17 samples tested positive for LT-ETEC, whereas V. cholerae, Shigella, and Salmonella were isolated in 91, 2, and 1 patient, respectively. Similar features such as acute watery diarrhea and dehydration were common to patients examined in the interepidemic period and those examined during the cholera epidemic. The serotypes isolated were O15 (8), O63 (2), O148 (2), and O158 (2), and the remaining strains were untypable. More than 50% of patients were adults. Antimicrobial susceptibility was as follows: amoxycillin (3/20), nalidixic acid (6/20), trimethoprim (5/20), chloramphenicol (8/20), amikacin (19/20), gentamicin (13/20), cefotaxime (15/20), and ciprofloxacin (10/20). High fluoroquinolone resistance in ETEC may be a serious cause of concern for travelers visiting this region, and thus thereis a need to monitor drug resistance in this pathogen.     

Genomic characterization of non-O1, non-O139 Vibrio cholerae reveals genes for a type III secretion system

Non-O1, non-O139 Vibrio cholerae can cause gastroenteritis and extraintestinal infections, but, unlike O1 and O139 strains of V. cholerae, little is known about the virulence gene content of non-O1, non-O139 strains and their phylogenetic relationship to other pathogenic V. cholerae. Comparative genomic microarray analysis of four pathogenic non-O1, non-O139 strains indicates that these strains are quite divergent from O1 and O139 strains. Genomic sequence analysis of a non-O1, non-O139 strain (AM-19226) that appeared particularly pathogenic in experimental animals suggests that this strain carries a type III secretion system (TTSS) that is related to the TTSS2 gene cluster found in a pandemic clone of Vibrio parahaemolyticus. The genes for this V. cholerae TTSS system appear to be present in many clinical and environmental non-O1, non-O139 strains, including at least one clone that is globally distributed. We hypothesize that the TTSS present in some pathogenic strains of non-O1, non-O139 V. cholerae may be involved in the virulence and environmental fitness of these strains.     

Surveillance of various enteropathogenic bacteria from diarrheal cases during 1989-1999 in Kobe City]

Incidence of various enteropathogenic bacteria was examined from diarrheal faecal samples that were collected from the patients of Kobe City General Hospital and some station hospitals (23,862), and from overseas travelers (2,855) over a period of decade (1989-1999) in Kobe. A total of 1,580 strains were isolated from domestic and 331 strains from overseas travelers. The results are as follows. 1) Thirteen kinds of enteropathogenic bacteria were isolated from domestic diarrheal cases (6.6%). Salmonella was the most predominant bacteria followed by Campylobacter, Vibrio parahaemolyticus, enteropathogenic Escherichia coli and Shigella. 2) Eleven kinds of enteropathogenic bacteria were isolated from overseas diarrheal travelers (11.6%). The most frequently isolated species was Salmonella, followed by Vibrio parahaemolyticus, Shigella and Plesiomonas shigelloides. 3) Of Salmonella strains isolated from domestic and overseas diarrheal cases, serovar Enteritidis was the most predominant. Other frequent serovars in both cases were Typhimurium, Tennessee, Hadar, Infantis, Blockley and Montevideo. 4) Antibiotics resistant rate of the isolated Salmonella strains was 42.6% for domestic samples and 29.3% for overseas diarrheal cases. In domestic cases. Enteritidis was resistance to streptomycin only and the multiple antibiotic resistance was observed in Typhimurium serovars. In overseas samples, the multiple antibiotic resistance was seen in a few Typhimurium, Anatum and Blockley strains. 5) Among Shigella, S. sonnei was isolated from both domestic and overseas cases. The frequency of acquiring infection was the highest in India, followed by Indonesia, Thailand and Nepal. 6) With reference to the incidence of the members of the genus Vibrio, Aeromonas and Plesiomonas, V. parahaemolyticus were abundant from domestic samples where as V. parahaemolyticus, P. shigelloides, Vibrio cholerae non-O1 and Vibrio cholerae O1 were isolated more frequently from overseas samples. The frequency of acquiring infection was the highest in Thailand, followed by Indonesia and India.     

Seasonality and antimicrobial resistance pattern of Vibrio cholerae in a tertiary care hospital of North India

We retrospectively analysed the seasonal distribution of cholera and the antimicrobial resistance pattern of Vibrio cholerae isolates over a 5-year period from January 1998 to December 2002. Of 3213 stool specimens processed from 3213 patients with acute watery diarrhoea during this period, 431 samples (13.4%) were found positive for V. cholerae. There were 423 V. cholerae O1 biotype E1 Tor, 2 V. cholerae O139 and six isolates of non-O1 non-O139. The highest number of cholera cases occurred in May-June followed by July-August. Cases started appearing in April for all years except in the year 2002 when three cases occurred in the first week of March. A large number (90.25% strains) were resistant to at least one antibiotic.