Bullous cellulitis as an extraordinary manifestation of a Vibrio cholerae O1 Ogawa infection

@#Vibrio cholerae is a gram-negative bacterium synonymous with its namesake disease, cholera. Thus, gastrointestinal symptoms are the norm and V. cholerae is very rarely associated with skin and soft tissue infections. We describe a case of a 63-year-old Chinese woman with multiple medical comorbidities on corticosteroid therapy who developed fever and a painful swelling on her left leg after being pricked by a branch while gardening. There was no abdominal pain, vomiting or diarrhea. A diagnosis of bullous cellulitis was made clinically, and blood was sent for bacteriological culture. A beta-hemolytic commashaped gram-negative bacillus was isolated from the blood. It was also oxidase-positive and produced an acid/alkaline (A/K) reaction on triple sugar iron agar. It was identified biochemically as Vibrio cholerae. After additional testing, it was found to be of the O1 serogroup and Ogawa serotype. The infection resolved following a 10-day course of high-dose co-trimoxazole therapy.

Expression in Saccharomyces boulardii of recombinant Toxin-coregulated pilus A subunit (TcpA) of Vibrio cholerae O1

Cholera is endemic in over 50 countries with an estimated mortality of 100,000-120,000. Vaccination is considered the complementary key to prevent and control cholera; therefore, alternative vaccine preparations are needed. Toxin Co-regulated Pilus is part of the toxR virulence regulon, which is necessary for colonization in the intestinal mucosa. In order to express Vibrio cholerae TcpA protein in Saccharomyces boulardii, the expression plasmid pYES2 was constructed by inserting tcpA gene isolated from local Vibrio cholerae Eltor Inaba isolates. The new construct was transferred into Saccharomyces boulardii cells and the expression of tcpA gene was induced from the GAL1 promoter by adding galactose to the medium. The SDS-PAGE and Western blot analysis showed the presence of TcpA in yeast. These results showed that Saccharomyces boulardii is a promising host to express Vibrio cholerae toxin TcpA as the first step in attempt to produce an oral Vibrio cholerae vaccine(AU)

El cólera es endémico en más de 50 países. Se estima una mortalidad entre 100.000 - 120.000 debido a esta enfermedad. La vacunación se considera una medida complementaria para prevenir y controlar el cólera, por lo tanto, se necesitan preparaciones vacunales alternativas a las existentes. El Pili corregulado con la toxina, es parte del regulón de virulencia toxR, y es necesario para la colonización en la mucosa intestinal. Para expresar la proteína tcpA de Vibrio cholerae en Saccharomyces boulardii, se construyó el plásmido de expresión pYES2 insertando el gen tcpA obtenido a partir de aislamientos locales de Vibrio cholerae El Tor Inaba. La nueva construcción se transfirió a las células de Saccharomyces boulardii y se indujo la expresión del gen tcpA a partir del promotor GAL1 mediante la adición de galactosa al medio. El análisis mediante SDS-PAGE y Western blot demostró la presencia de TcpA en levaduras. Los resultados demostraron que Saccharomyces boulardii es un hospedero prometedor para expresar el gen tcpA de Vibrio cholerae como el primer paso en el intento de producir una vacuna oral contra Vibrio cholerae(AU)

Comparison of colonization ability of O1 and O139 Vibrio cholerae strains on soft-shelled turtle's surface / 中华预防医学杂志

Objective@#To study the preferred colonization sites of O1 Vibrio cholerae (V.cholerae) and the colonization ability difference for O1 and O139 V. cholerae on soft-shelled turtle's surface.@*Methods@#8 O1 and O139 V. cholerae strains were obtained from branch of diarrheal diseases, Chinese center for disease control and prevention. 63 soft-shelled turtles weighing 150 g and 9 cm in length (diameter of calipash) were selected for use in the study. The preferred colonization sites and proliferation trend were studied by using bioluminescent imaging method. The colonization factors for O1 V. cholerae strains were studied by constructing colonization gene mutant strains (VC1897dmshA, VC1897dgbpA and VC1897dtcpA), performing competition colonization assays and analyzing the competitive indexes. After pairing off O1 and O139 strains respectively to perform 16 competition groups, the colonization difference of these two strains were studied by competition colonization assays.@*Results@#The colonization sites by V. cholerae on soft-shelled turtles surfaces was clustered. More V. cholerae strains colonized on turtle's calipash and carapace on dorsal side and less strain colonized on ventral side. The competition colonization assays showed that colonization ability of O1 serogroup mshA mutant strains were 7.26 times lower than VC1897dlacZ. Besides, the CI value (O139/O1) of 11 out of the 16 competition groups were greater than 2 (between 2.07 and 59.84). Two groups showed values of 1.43 and 0.93 respectively and 3 groups lower than 0.7.@*Conclusion@#The preferred colonization sites for O1 V. cholerae strains on body surface were observed.MSHA was one of the main colonization factors for its colonization. Our study suggested that in general, O139 V. cholerae strains have stronger colonization ability than O1 strains. Besides, strains isolated from soft-shelled turtles tend to have stronger colonization ability than strains isolated from patients.

Susceptibilidad antimicrobiana en aislamientos cubanos de Vibrio cholerae O1 procedentes de muestras clínicas / Antimicrobial susceptibility in Cuban isolates of Vibrio cholerae O1 obtained from clinical samples

Introducción: la re-emergencia de cólera en Haití estableció un nuevo reservorio para el incremento de la séptima pandemia. Esto provocó su diseminación a República Dominicana y a otros países de la región del Caribe, como Cuba y México. Objetivo: estudiar la susceptibilidad antimicrobiana de aislamientos de Vibrio cholerae O1, serotipo Ogawa, biotipo El Tor aisladas de pacientes durante el evento epidemiológico de cólera ocurrido en Cuba entre junio de 2012 y agosto de 2013. Métodos: se realizó el estudio de la susceptibilidad antimicrobiana in vitro en 144 aislamientos de V. cholerae, mediante el método de Bauer-Kirby frente a nueve antimicrobianos: ampicilina, sulfonamida, trimetoprim/sulfametoxazol, cloranfenicol, tetraciclina, doxiciclina, azitromicina, ciprofloxacina y gentamicina, según las normas del Instituto de Estándares de Laboratorio Clínico de los Estados Unidos de América. Resultados: el total de los aislamientos resultaron resistentes al trimetoprim-sulfametoxazol; el 98,7 por ciento lo fue a la sulfonamida y el 90,3 por ciento a la ampicilina. Se obtuvieron valores de sensibilidad intermedia para ciprofloxacina (30,6 por ciento) y cloranfenicol (27,1 por ciento). Se apreciaron niveles de sensibilidad superior al 92 por ciento a los antimicrobianos de primera línea en el tratamiento de la enfermedad (doxiciclina, tetraciclina y azitromicina), así como también a la gentamicina. No se observaron cepas multirresistentes. Conclusiones: los datos aportados por este trabajo demuestran la efectividad in vitro de los antimicrobianos utilizados en el tratamiento la enfermedad diarreica aguda causada por V. cholerae en Cuba(AU)

Introduction: re-emergence of cholera in Haiti created a new reservoir for the increase of the seventh pandemic. This resulted in its spread to the Dominican Republic and other Caribbean countries, such as Cuba and Mexico. Objectives: study the antimicrobial susceptibility of isolates of Vibrio cholerae O1, Ogawa serotype, El Tor biotype, obtained from patients during the cholera epidemiological event occurring in Cuba from June 2012 to August 2013. Methods: a study was conducted of 144 V. cholerae isolates using the Bauer-Kirby method to determine in vitro susceptibility to nine antimicrobials: ampicillin, sulfonamide, trimethoprim/sulfamethoxazole, chloramphenicol, tetracycline, doxycycline, azithromycin, ciprofloxacin and gentamicin, in compliance with standards from the U.S. Clinical and Laboratory Standards Institute. Results: all isolates were resistant to trimethoprim/sulfamethoxazole; 98.7 percent to sulfonamide and 90.3 percent to ampicillin. Intermediate sensitivity values were obtained for ciprofloxacin (30.6 percent) and chloramphenicol (27.1 percent). Sensitivity levels above 92 percent were found for first-line antimicrobials (doxycycline, tetracycline and azithromycin), as well as gentamicin. Multi-drug resistant strains were not found. Conclusions: results reveal the effectiveness in vitro of the antimicrobials used in Cuba to treat acute diarrheal disease caused by V. cholerae(AU)

Vibrio cholerae O1 from superficial water of the Tucunduba Stream, Brazilian Amazon

Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized.

Vibrio cholerae O1 from superficial water of the Tucunduba Stream, Brazilian Amazon

Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized.

Significância de anticorpos vibriocidas circulantes em área pós-epidêmica de diarréia, São Bento do Una, Estado de Pernambuco / The importance of circulating vibriocidal antibodies in a pos-epidemic region of diarrhea, São Bento do Una, State of Pernambuco

Verificou-se o nível de anticorpos vibriocidas em 41 indivíduos adultos, sem história passada ou presente de diarréia por Vibrio cholerae O1, residentes no município de São Bento do Una, Pernambuco. Nessa localidade ocorreu no início de 2004 um surto de diarréia, com múltiplos agentes bacterianos envolvidos, incluindo o vibrião colérico. Foi empregado o teste da microtitulação de anticorpos séricos vibriocidas, anti-Ogawa e anti-Inaba, considerando-se como indicativo de infecção por Vibrio cholerae O1, os títulos vibriocidas > 1:640. A freqüência dos reagentes foi de 36 (87,8 por cento) para o sorovar Ogawa, o que evidencia a possível circulação do vibrião colérico, durante e/ou após a epidemia de diarréia.

The levels of vibriocidal antibodies were investigated among 41 adults without any past or present history of diarrhea due to Vibrio cholerae O1 who were living in the municipality of São Bento do Una, Pernambuco. A diarrhea outbreak occurred in this locality at the beginning of 2004, involving multiple bacterial agents, including Vibrio cholerae. The microtitration test was used to investigate the presence of anti-Ogawa and anti-Inaba vibriocidal serum antibodies. Vibriocidal titers e" 1:640 were considered indicative of infection by Vibrio cholerae O1. The frequency of the reagents was 36 (87.8 percent) for the Ogawa serovar, which showed that Vibrio cholerae O1 was possibly circulating during and/or after the diarrhea epidemic.

Detection of viable and viable nonculturable Vibrio cholerae O1 through cultures and immunofluorescence in the Tucumán rivers, Argentina

Vibrio cholerae has been sporadically isolated from rivers in Tucumán, Argentina, since the outbreak in 1991. The aim of this study was to determine the environmental reservoir of the bacterium in these rivers, assessing the presence of Vibrio cholerae non-O1 and O1 (the latter both in its viable culturable and non culturable state) and its relationship to environmental physicochemical variables. 18 water samplings were collected in the Salí River (in Canal Norte and Banda) and the Lules River between 2003 and 2005. Physical-chemical measurements (pH, water temperature, electrical conductivity and dissolved oxygen) were examined. Vibrio cholerae was investigated with conventional culture methods and with Direct Immunofluorescence (DFA-VNC) in order to detect viable non culturable organisms. All isolated microorganisms corresponded to Vibrio cholerae non-O1 and non-O139 (Lules 26 percent, Canal Norte 33 percent and Banda 41 percent). The majority was found during spring and summer and correlated with temperature and pH. Non culturable Vibrio cholerae O1 was detected year round in 38 of the 54 water samples analyzed. Application of the Pearson correlation coefficient revealed that there was no relationship between positive immunofluorescence results and environmental physicochemical parameters. Genes coding for somatic antigen O1 were confirmed in all DFA-VNC-positive samples, whereas the virulence-associated ctxA and tcpA genes were confirmed in 24 samples.

Vibrio cholerae tem sido isolado esporadicamente nos rios da Província de Tucumán, Argentina, desde outubro de 1991. O objetivo deste estudo foi localizar os reservatórios nestes rios, identificar a presença de Vibrio cholerae O1 (em estado cultivável e não cultivável) e relacionar a presença desta bactéria com as variações físico-químicos da água. Foram coletadas dezoito amostras de água do rio Salí (nas localidades de Canal Norte e Banda) e do rio Lules, entre 2003 e 2005. Estas foram submetidas a análises físico-químicos como determinação de pH, temperatura, condutibilidade elétrica e oxigênio dissolvido. A presença de Vibrio cholerae foi verificada por métodos de cultivo convencional e por imunofluorescência direta (DFA-VNC). Todos os microrganismos isolados foram não O1 e não O139 (Lules 26 por cento, Canal Norte 33 por cento e Banda 41 por cento). A maioria foi encontrada na primavera e verão, indicando uma relação com a temperatura e pH. Das 54 amostras analisadas por DFA-VNC, 38 Vibrio cholerae não cultivável, foram detectadas em todas as épocas do ano. As amostras positivas foram confirmadas por PCR para o antígeno somático O1 e para os genes de virulência ctxA e tcpA. Coeficiente de correlação de Pearson revelou que não há relação entre os resultados obtidos por imunofluorescência e a variação dos parâmetros físico-químicos.