Serotypes and phage types of Vibrio cholerae in Mysore.

A total of 565 samples of acute diarrhoeal stools from children received over a period of twelve months from November 1996 to October 1997, yielded 289 (51.15%) isolates of V. cholerae. Out of these, 277 (95.85%) were V. cholerae 01 serotype Ogawa, 7 (2.42%) were V. cholerae 0139 and 5 (1.73%) were V. cholerae non-O1 non-0139. Phage typing of the isolates was carried out by NICED Calcutta. Out of 78 isolates, 69 (88.46%) belong to phage type T27. Of the remaining, 3 (3.85%), 3(3.85%), 2(2.56%) and 1(1.28%) belong to the phage types T26, T13, T8 and T4 respectively.     

Non-cholera vibrios in enterobacteriologic routine]

Of 3250 diarrheal stools received for microbiologic diagnosis at a private clinical laboratory in Recife, Brazil, strains of Vibrio were isolated from 55 (1.7%). The study was carried out from May 1989 through May 1991. For recovering Vibrio, fecal samples were enriched in alkaline peptone water supplemented with 2% NaCl and subcultured on thiosulfate-citrate-bile salts-sucrose agar (TCBS). Of the recovered species, V. parahaemolyticus was most commonly found (24 strains), followed by V. furnissii (15 strains), V. cholerae non-01 (6 strains), V. alginolyticus (4 strains), V. fluvialis (2 strains), and Vibrio sp. (1 strain). The low isolation rate of Vibrio raises doubts about the cost-effectiveness of the use of TCBS in the routine enterobacteriologic workup of clinical laboratories.     

The characterization of Vibrio cholerae isolated in Kenya in 1983

A total of 245 strains of Vibrio cholerae 01 and two strains of V. cholerae non-01 were isolated and collected from diarrhoeal patients in Homa Bay District Hospital and the other medical facilities in Nyanza Province, Kenya in 1983. The majority of V. cholerae 01 tested were Ogawa type (with the exception of nine Inaba type), biotype E1 Tor (except one untypable strain) and Celebes original type (except one cured type). Haemolytic activity to sheep red blood cells was detected in 75.5% of isolates. Out of 245 strains of V. cholerae 01, 184 were resistant to tetracycline, streptomycin and ampicillin. All were sensitive to chloramphenicol and nalidixic acid. Only one strain of V. cholerae 01 was sensitive to all five antimicrobial agents tested. An environmental cholera survey was done after the cholera outbreak subsided. Twenty strains of V. cholerae non-01 were isolated from water samples in Nyanza Province but none of V. cholerae 01 was isolated.     

Outbreak of cholera associated with crab brought from an area with epidemic disease.

From 31 March through 3 April 1991, 8 New Jersey residents developed severe, watery diarrhea after eating crabmeat brought back in the suitcase of a traveler to Ecuador. Stool cultures yielded toxigenic Vibrio cholerae O1, serotype Inaba, biotype El Tor from 4 persons, and vibriocidal antibody titers were > or = 1:640 in 7 persons, indicating recent infection with Vibrio cholerae O1. Eating crab was statistically associated with illness (P = .006); however, no leftover crabmeat was available for testing. All 8 patients fully recovered and no cases of secondary transmission were reported. This was the first reported incident of cholera in the continental United States associated with food transported from an area with epidemic disease. Discouraging the transport of perishable souvenir seafood may prevent further outbreaks.     

A comparative study of enterotoxigenic Escherichia coli, Shigella, Aeromonas, and Vibrio as etiologies of diarrhea in northeastern Thailand

The incidence of enterotoxigenic Escherichia coli (ETEC), Shigella, Aeromonas, and Vibrio was determined in patients with diarrhea seen at a hospital in northeastern Thailand, and compared with the incidence of these bacteria in household contacts and their neighbors. ETEC was identified in 17%, Shigella in 9%, Aeromonas in 9%, V. parahaemolyticus in 5%, and non-01 V. cholerae in 2% of 299 patients with diarrhea. These five species of bacteria were isolated more often from patients with diarrhea than persons without diarrhea (P less than 0.001). ETEC was found more often in household contacts (22/141) and neighbors (18/147) of index cases than in persons living in homes not associated with ETEC infections (32/1,318; P less than 0.001). While Shigella was isolated less often in family contacts (3/76) and neighbors (4/93) of patients with shigellosis, this enteric pathogen was also isolated more often from contacts than persons not associated with Shigella infection (13/1,437; P less than 0.001). Both Aeromonas and non-01 V. cholerae can also be enteric pathogens; further efforts should be made to define the enteropathogenicity of these bacteria.     

Toxigenic Vibrio cholerae serogroup O141-associated cholera-like diarrhea and bloodstream infection in the United States

Toxigenic Vibrio cholerae serogroup O141 has been associated with sporadic cholera-like diarrhea and bloodstream infection in the United States. Consumption of seafood and proximity to the coast may increase the risk of infection. All V. cholerae isolates recovered from stool samples of patients with diarrhea or from a normally sterile site should be serogrouped and assessed for cholera toxin production. Improved surveillance and case-control studies are needed to further characterize illness and risk factors for V. cholerae O141 infection.     

Complexity of rice-water stool from patients with Vibrio cholerae plays a role in the transmission of infectious diarrhea

At the International Centre for Diarrhoeal Disease Research, Bangladesh, one-half of the rice-water stool samples that were culture-positive for Vibrio cholerae did not contain motile V. cholerae by standard darkfield microscopy and were defined as darkfield-negative (DF(-)). We evaluated the host and microbial factors associated with DF status, as well as the impact of DF status on transmission. Viable counts of V. cholerae in DF(-) stools were three logs lower than in DF(+) stools, although DF(-) and DF(+) stools had similar direct counts of V. cholerae by microscopy. In DF(-) samples, non-V. cholerae bacteria outnumbered V. cholerae 10:1. Lytic V. cholerae bacteriophage were present in 90% of DF(-) samples compared with 35% of DF(+) samples, suggesting that bacteriophage may limit culture-positive patients from producing DF(+) stools. V. cholerae in DF(-) and DF(+) samples were found both planktonically and in distinct nonplanktonic populations; the distribution of organisms between these compartments did not differ appreciably between DF(-) and DF(+) stools. This biology may impact transmission because epidemiological data suggested that household contacts of a DF(+) index case were at greater risk of infection with V. cholerae. We propose a model in which V. cholerae multiply in the small intestine to produce a fluid niche that is dominated by V. cholerae. If lytic phage are present, viable counts of V. cholerae drop, stools become DF(-), other microorganisms bloom, and cholera transmission is reduced.     

Vibrios and Aeromonas

There are many similarities in the Vibrionaceae that cause human illness in the United States (see Table 1). Vibrios are characteristically indigenous to marine, estuarine, and brackish environments. They are distributed mainly in Gulf of Mexico coastal water, and these organisms "bloom" when the water is warm. Outbreaks of disease in humans frequently occur in summer, coinciding with multiplication of vibrios in warm water. Sporadic cases and small outbreaks of cholera continue to occur in persons living on or near the Gulf of Mexico, but infection in most persons is unrecognized. In fact, more serious and frequent illnesses result from V. vulnificus wound infections and from gastroenteritis caused by vibrios other than V. cholerae 01. Underlying hepatic or neoplastic disease (especially leukemia) apparently increases the likelihood and severity of illnesses caused by V. vulnificus and Aeromonas. Some Vibrionaceae produce clinical illness by means of enterotoxins identical or similar to cholera toxin. For many others, hemolysins, cytotoxins, and other exotoxins are necessary to produce disease; the importance of these virulence factors often is not known or the importance of these virulence factors often is not known or is of doubtful significance. Also, purported pathogenicity as demonstrated by animal models, such as fluid accumulation in ligated ileal loops, is quite nonspecific and needs to be interpreted cautiously. For Plesiomonas, a mode of pathogenesis has not been discovered. Eating raw shellfish (frequently raw oysters) has been linked epidemiologically to enteric infections with most of these bacteria; foreign travel and exposure to seawater are other frequently observed epidemiologic associations with infection. Foreign travel, particularly to the Yucatan Peninsula of Mexico, has been strongly associated with the acquisition of non-01 V. cholerae and Plesiomonas organisms. Most Vibrionaceae in the United States are susceptible in vitro--and illnesses from them are responsive--to tetracycline, trimethoprim-sulfamethoxazole, and other common antimicrobial agents. However, as for other bacteria that cause diarrhea, the main treatment for uncomplicated disease is the judicious replacement of fluids and electrolytes lost in diarrhea. A loose network of surveilance for these organisms comprises hospital and public health laboratories in Gulf coastal states that plate diarrheal stools on TCBS agar. As recognized pathogens are more assiduously screened for, and as newly identified vibrios are definitely included or excluded as enteric pathogens, the clinical importance of these members of the Vibrionaceae family should become clearer.     

Transmissibility of cholera: in vivo-formed biofilms and their relationship to infectivity and persistence in the environment

The factors that enhance the waterborne spread of bacterial epidemics and sustain the epidemic strain in nature are unclear. Although the epidemic diarrheal disease cholera is known to be transmitted by water contaminated with pathogenic Vibrio cholerae, routine isolation of pathogenic strains from aquatic environments is challenging. Here, we show that conditionally viable environmental cells (CVEC) of pathogenic V. cholerae that resist cultivation by conventional techniques exist in surface water as aggregates (biofilms) of partially dormant cells. Such CVEC can be recovered as fully virulent bacteria by inoculating the water into rabbit intestines. Furthermore, when V. cholerae shed in stools of cholera patients are inoculated in environmental water samples in the laboratory, the cells exhibit characteristics similar to CVEC, suggesting that CVEC are the infectious form of V. cholerae in water and that CVEC in nature may have been derived from human cholera stools. We also observed that stools from cholera patients contain a heterogeneous mixture of biofilm-like aggregates and free-swimming planktonic cells of V. cholerae. Estimation of the relative infectivity of these different forms of V. cholerae cells suggested that the enhanced infectivity of V. cholerae shed in human stools is largely due to the presence of clumps of cells that disperse in vivo, providing a high dose of the pathogen. The results of this study support a model of cholera transmission in which in vivo-formed biofilms contribute to enhanced infectivity and environmental persistence of pathogenic V. cholerae.     

Cholera in the United States, 1995-2000: trends at the end of the twentieth century

To evaluate recent trends in cholera in the United States, surveillance data from all cases of laboratory-confirmed toxigenic Vibrio cholerae O1 and O139 infection reported to the Centers for Disease Control and Prevention between 1995 and 2000 were reviewed. Sixty-one cases of cholera, all caused by V. cholerae O1, were reported. There was 1 death, and 35 (57%) of the patients were hospitalized. Thirty-seven (61%) infections were acquired outside the United States; 14 (23%) were acquired through undercooked seafood consumed in the United States, 2 (3%) were acquired through sliced cantaloupe contaminated by an asymptomatically infected food handler, and no source was identified for 8 (13%) infections. The proportion of travel-associated infections resistant to trimethoprim-sulfamethoxazole, sulfisoxazole, streptomycin, and furazolidone increased from 7 (8%) of 88 in 1990-1994 to 11 (31%) of 35 in 1995-2000. Foreign travel and undercooked seafood continue to account for most US cholera cases. Antimicrobial resistance has increased among V. cholerae O1 strains isolated from ill travelers.     

Non-O1 Vibrio cholerae Bacteremia in Patients with Cirrhosis: 5-yr Experience from a Single Medical Center

Objectives: To assess the clinical features and susceptibility of cirrhotic patients to non-01 Vibrio cholerae bacteremia and to provide our therapeutic experiences in this rare and highly lethal infection. Methods: Twenty-eight blood culture isolates of non-01 V. cholerae were identified by our clinical microbiology laboratory between July 1989 and June 1994. Patients with underlying cirrhosis and the aforementioned bacteremia were retrospectively reviewed. Results: Twenty-one cirrhotic patients (16 male, five female; mean age, 50.9 yr; range 28–67 yr) were identified and classified as Child B (6 cases) and Child C (15 cases). Bacteremic episodes occurred most often from March to September. Seafood ingestion (seven cases) and seawater exposure (two cases) were risk factors, but nosocomial infections were also noted in six cases. Presenting symptoms and signs included ascites (95.2%), fever (81%), abdominal pain (52.4%), diarrhea (33.3%), and cellulitis with bullae Formation (19%). Concurrent spontaneous bacterial peritonitis was determined in 10 cases, seven with positive ascites cultures. Antibiotic therapy (either cephalothin with gentamicin or ceftriaxone alone) cured most of the bacteremic episodes. The overall case-fatality rate was 23.8%, hut 75% of the deaths were observed in patients with skin manifestation. Conclusions: Patients with decompensated cirrhosis are susceptible to non-01 V. cholerae bacteremia and should not ingest raw sea food or expose skin wounds to salt water. A high index of suspicion and early administration of antibiotics may lower the mortality rate.     

Emergence of Vibrio cholerae O1 biotype El Tor serotype Inaba causing outbreaks of cholera in Orissa, India

A total of 431 rectal swabs, collected from acute diarrheal cases at a surveillance site and at different diarrheal outbreak areas of Orissa from May to October 2005, were bacteriologically analyzed. Out of 265 culture-positive samples, Vibrio cholerae O1 was isolated in 56 samples (20.8%), of which 37 were the Inaba serotype and 19 were the Ogawa. The antibiogram profile revealed that all the V. cholerae O1 Ogawa and Inaba serotypes were uniformly sensitive to ampicillin, chloramphenicol, gentamicin, ciprofloxacin, norfloxacin and tetracycline. The V. cholerae O1 Inaba serotypes were resistant to furazolidone and nalidixic acid, while the Ogawa strains were resistant to furazolidone, nalidixic acid and neomycin. The multiplex polymerase chain reaction (PCR) assay on some selected strains of both serotypes revealed that all the strains were positive for ctxA and tcpA genes showing biotype El Tor. The present study revealed the emergence of V. cholerae O1 biotype El Tor serotype Inaba, which caused sporadic outbreaks of cholera in 2005. The outbreaks of diarrheal disorders in one geographical area of the state (in the Pattamundai area, Kendrapara district) in 2005 were due to V. cholerae O1 Ogawa, whereas the other outbreaks in other areas (Puri, Khurda and Dhenkanal districts) from August to October 2005 were due to V. cholerae O1 serotype Inaba. This is the first report that an emergence of V. cholerae O1 serotype Inaba caused sporadic outbreaks of cholera in different parts of Orissa. Switching over of V. cholerae O1 Ogawa strains to Inaba, causing diarrheal outbreaks in Orissa, needs close monitoring.     

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.     

Attachment of toxigenic Vibrio cholerae 01 to various freshwater plants and survival with a filamentous green alga, Rhizoclonium fontanum

In Bangladesh, cholera epidemics occur twice a year. V. cholerae 01 are readily isolated from the environment only during epidemics. The interepidemic reservoirs or sites of survival and multiplication of V. cholerae are still unknown. Investigations were carried out with various fresh-water plants as possible reservoirs of V. cholerae in the environment. Attachment to and acute population changes of V. cholerae on various plant surfaces was used as a screening technique to screen a particular plant species for survival studies. Five plant species, Anabaena variabilis, Rhizoclonium fontanum, Cladophora sp., Fontinalis antipyretica and Elodea canadensis were used for attachment experiments. Among these plants, R. fontanum showed the best attachment. On the basis of attachment results, survival experiments were carried out with R. fontanum. At 0.05% salinity toxigenic V. cholerae 01 survive longer in the presence of R. fontanum than in medium without algae.     

Molecular tracking of the lineage of strains of Vibrio cholerae O1 biotype El Tor associated with a cholera outbreak in Andaman and Nicobar Islands, India

A large outbreak of acute watery diarrhoea involving all age groups of mongoloid tribal aborigines occurred during October-November, 2002 in the Nancowry group of Andaman and Nicobar Islands in the Indian Ocean. Twenty-one of the 67 stool samples from 67 patients were positive for toxigenic Vibrio cholerae O1, serotype Ogawa biotype El Tor, which showed striking similarity in its antibiogram with some of the strains of V. cholerae O1 Serotype Ogawa biotype El Tor isolated in Kolkata. The Nancowry and Kolkata isolates were compared with molecular tools involving random amplified polymorphic DNA (RAPD) fingerprinting, ribotyping and pulsed-field gel electrophoresis (PFGE). RAPD fingerprinting and ribotyping techniques revealed that all the V. cholerae strains associated with the outbreak in these islands were clonal in nature and identical to a population of isolates obtained from Kolkata since 1993. PFGE could discriminate within these Kolkata isolates further and established that a particular subtype of this population reached the remote Nancowry islands and was responsible for the outbreak.     

Vector potential of houseflies (Musca domestica) in the transmission of Vibrio cholerae in India

It is well known that diarrhoeal infections due to Vibrio cholerae are spread through fecal-oral route of transmission. In the present study an attempt was made to isolate and identify V. cholerae from houseflies, Musca domestica collected from a low socioeconomic area in Delhi, India, where an outbreak of cholera was encountered. Of the ten fly pools examined, six (60%) were positive for V. cholerae. Of these six pools, three (50%) showed V. cholerae Ogawa T2 El Tor and one (17.5%) V. cholerae non-O1. Two isolates could not be typed. During the outbreak period, V. cholerae Ogawa T2 El Tor was isolated from stools of patients suffering from diarrhoea. These findings suggest that houseflies act as mechanical vectors of V. cholerae biotype El Tor and may help in their dissemination. The present study highlights the recovery of V. cholerae El Tor from M. domestica which, to the authors knowledge, has not been reported previously.     

Clinical studies in persistent diarrhea: dietary management with green banana or pectin in Bangladeshi children

BACKGROUND & AIMS: Because of the beneficial intestinal effects of dietary fibers, we have evaluated the therapeutic effects of green banana or pectin in children with persistent diarrhea. METHODS: In a double-blind trial, 62 boys, age 5-12 months, were randomly given a rice-based diet containing either 250 g/L of cooked green banana (n = 22) or 4 g/kg pectin (n = 19) or the rice-diet alone (control, n = 21), providing 54 kcal/dL daily for 7 days. Stool weight and consistency, frequency of vomiting and purging, and duration of illness were measured. RESULTS: Most children (60%) had no pathogens isolated from stools, 17% had rotavirus, 5% Vibrio cholerae, 4% Salmonella group B, and 11% had enterotoxigenic Escherichia coli infections. By day 3 posttreatment, significantly (P < 0.001) more children recovered from diarrhea receiving pectin or banana than controls (59%, 55%, and 15%, respectively). By day 4, these proportions correspondingly increased to 82%, 78%, and 23%, respectively, the study diet groups being significantly (P < 0.001) different than controls. Green banana and pectin significantly (P < 0.05) reduced amounts of stool, oral rehydration solution, intravenous fluid, and numbers of vomiting, and diarrheal duration. CONCLUSIONS: Green banana and pectin are useful in the dietary management of persistent diarrhea in hospitalized children and may also be useful to treat children at home.     

Seasonal, nontoxigenic Vibrio cholerae O1 Ogawa infections in the Eastern Region of Saudi Arabia.

BACKGROUND: Surveillance for Vibrio cholerae in the Eastern Region of Saudi Arabia has been ongoing since 1985 to detect and prevent local proliferation of imported cholera. In 1996 and 1997 the authors performed additional microbiologic and epidemiologic assessment of V. cholerae surveillance to better characterize a recurrent summertime pattern of V. cholerae infections in the Eastern Region of Saudi Arabia. METHODS: All health facilities routinely submitted stool or rectal swab specimens for isolation of V. cholerae from patients with gastroenteritis. In addition, specimens were taken from expatriate workers and household contacts of persons with confirmed V. cholerae infection. Forty-two isolates were evaluated for cholera enterotoxin by enzyme-linked immunosorbent assay, cholera toxin polymerase chain reaction, and Y1 adrenal cell assay; 12 isolates also were characterized by pulsed-field gel electrophoresis (PFGE). Interviews about potential exposures were done for all V. cholerae infections. RESULTS: Vibrio cholerae O1 serotype Ogawa biotype El Tor was identified in 113 gastroenteritis patients (6.0 per 100,000 population per year), 28 asymptomatic expatriate workers, and 16 of 982 household contacts of index patients. All symptomatic infected persons had mild illness that was not typical of cholera, and all 42 isolates evaluated were nontoxigenic. All 12 isolates evaluated by PFGE had an indistinguishable pattern (pattern 81). Infections appeared in late May, decreased in mid-July through August, increased again in September, and disappeared from December through April. Infections had a uniform geographic distribution and affected all ages. No linkage was identified between affected households, or between community cases and food-handlers or domestic servants. DISCUSSION: Surveillance in the Eastern Region of Saudi Arabia has identified a novel strain of nontoxigenic V. cholerae O1 Ogawa. This strain probably has a local environmental reservoir. Since cholera toxin is the primary virulence factor involved in the cause of cholera, assays for cholera toxin should be included in cholera surveillance.     

Surveillance of bacterial pathogens associated with acute diarrhea in Lima, Peru.

OBJECTIVES: A study was conducted in Lima, Peru, from January to April 1995, to determine the bacterial pathogens associated with acute diarrhea in adults, their susceptibility to common antimicrobials, the risk factors involved in cholera transmission, and the best clinical predictors of cholera. METHODS: A random sample of adult patients with acute diarrhea was studied. Epidemiologic and clinical data and risk factors to acquire diarrheal diseases were evaluated. Identification of bacteria and susceptibility to antimicrobials were determined. RESULTS: The study included 336 patients. Vibrio cholerae O1 (52.7%), Shigella spp. (4. 8%), and Salmonella spp (2.7%) were the pathogens most commonly isolated. No resistance to antimicrobials was observed. Patients with cholera had less access to municipal water (P = 0.0018) and were less likely to have homes connected to a sewage system (P = 0. 0003) or to have indoor toilet facilities (P = 0.0001) than those without cholera. Liquid stools (odds ratio [OR] = 16.51; confidence interval [CI] = 13.71-19.02; P = 0.003), severe dehydration (OR = 2. 48; CI = 1.57-3.38; P = 0.0083), generalized cramps (OR = 4.63; CI = 3.10-6.17, P < 0.0001), and washerwoman's hands (OR = 2.45; CI = 1. 55-3.34; P = 0.017) were the best clinical predictors of cholera in this setting. CONCLUSIONS: Cholera is still prevalent in Lima, and people living in environments with low sanitary conditions are especially at risk. Clinical signs of severe dehydration and liquid stools were the best predictors of cholera.     

食品与临床分离的致病性副溶血性弧菌耐药性比较

目的比较食品与临床分离的致病性副溶血性弧菌耐药性差异,为该菌耐药机制研究提供基础。方法运用K-B纸片法,对上海市28株临床菌株、18株食品源菌株的副溶血性弧菌进行耐药性监测,以PCR分析菌株的耐药基因。再用多位点测序技术揭示46株副溶血性弧菌的遗传多样性,针对相同进化分支中不同来源的分离株进行耐药性比较。结果28株临床菌株的耐药率(100%)明显高于18株食品源分离株(88.9%),而且临床菌株全部为多重耐药性菌株(n=28),而食品源中仅有2株具有多重耐药性。临床菌株所携带的耐药基因数量比食品源分离菌株更多、种类更为丰富。多位点测序分型结果也显示相同的趋势,在同一进化分支中,临床菌株的耐药性也显著高于食品分离株。结论本研究系统地比较食品与临床分离的致病性副溶血性弧菌耐药表型与耐药基因型的差异,分析了耐药性形成的原因,为副溶血性弧菌耐药性的起源、传播与控制提供依据。