[Aeromonas spp asociated to acute diarrheic disease in Cuba: case-control study]. / Aeromonas spp asociada a enfermedad diarreica aguda en Cuba: estudios de casos y controles.

The members of the genus Aeromonas are currently considered important gastrointestinal pathogens in different geographical areas. From February 1985 to January 2005 several case-control studies were coordinated by the National Reference Laboratory for Diarrheal Diseases from the Pedro Kouri Institute. The study purpose was to analyze a possible pathogenic role for Aeromonas spp in Cuban children with acute diarrhea. In that period 2,322 children less than 5 years old with acute diarrhea were studied for diarhoeal pathogens and another group of 2,072 non hospitalized children without diarrhea during the similar time from the same geographical areas and matched by ages were recruited. In the group of children with diarrheas (cases), Aeromonas spp. was isolated in 166 (7.15%) and in the control group the microorganism was found in only 35 (1.76%). When Aeromonas isolation rates were compared between both groups, we found that probability to isolate this specie was significantly higher in cases than in controls (OR = 4.48, 95% IC: 3.05-6.60; P < 0.001). The Aeromonas species more frequently isolated were A. caviae, A. hydrophila, and A. veronii bv sobria. Other enteric pathogens detected in children with diarrhea were: Shigella spp in 418 (18%) (P < 0.0001), Salmonella spp in 53 (2.3%) (P < 0.01), and enteropathogenic E. coli in 58 (2.49%) (P < 0.05).

Aeromonas spp asociada a enfermedad diarreica aguda en Cuba: estudios de casos y controles / Aeromonas spp asociated to acute diarrheic disease in Cuba: case-control study

The members of the genus Aeromonas are currently considered important gastrointestinal pathogens in different geographical areas. From February 1985 to January 2005 several case-control studies were coordinated by the National Reference Laboratory for Diarrheal Diseases from the Pedro Kouri Institute. The study purpose was to analyze a possible pathogenic role for Aeromonas spp in Cuban children with acute diarrhea. In that period 2,322 children less than 5 years old with acute diarrhea were studied for diarhoeal pathogens and another group of 2,072 non hospitalized children without diarrhea during the similar time from the same geographical areas and matched by ages were recruited. In the group of children with diarrheas (cases), Aeromonas spp. was isolated in 166 (7.15%) and in the control group the microorganism was found in only 35 (1.76%). When Aeromonas isolation rates were compared between both groups, we found that probability to isolate this specie was significantly higher in cases than in controls (OR = 4.48, 95% IC: 3.05-6.60; P < 0.001). The Aeromonas species more frequently isolated were A. caviae, A. hydrophila, and A. veronii bv sobria. Other enteric pathogens detected in children with diarrhea were: Shigella spp in 418 (18%) (P < 0.0001), Salmonella spp in 53 (2.3%) (P < 0.01), and enteropathogenic E. coli in 58 (2.49%) (P < 0.05).

Los miembros del género Aeromonas son considerados patógenos importantes del tracto gastrointestinal en diferentes áreas geográficas. De febrero de 1985 a enero de 2005 se realizaron estudios de casos y controles en el Laboratorio Nacional de Referencia de Enfermedades Diarreicas Agudas del Instituto Pedro Kourí con el objetivo de conocer el comportamiento de los microorganismos pertenecientes al género Aeromonas en niños con diarreas en Cuba. La muestra estuvo constituida por 2.322 niños bajo 5 años de edad, ingresados por enfermedad diarreica aguda y como grupo control se estudió un total de 2.072 niños con edades y áreas geográficas similares que acudieron a los hospitales correspondientes en ese mismo período de tiempo. En el grupo de niños que presentaron diarreas, Aeromonas spp fue aislada en 166 (7,15%), y en los controles fue encontrada en 35 (1,76%). Al comparar la positividad para Aeromonas entre ambos grupos, la probabilidad de diagnosticar este microorganismo fue 4,28 veces mayor en los casos que en los controles (OR = 4,28; IC al 95%: 2,96-6,20; P < 0,001). Las especies más frecuentemente aisladas fueron A. caviae, A. hydrophila, y A. veronii bv sobria. Otros enteropatógenos aislados en niños que presentaban diarrea fueron: Shigella spp. en 418 (18%) (P < 0,0001, Salmonella spp en 53 (2,3%) (P < 0,01),) y E. coli enteropatógena en 58 (2,49%) (P < 0,05).

[Phenotypic characteristics and virulence factors in Aeromonas strains isolated from patients with diarrheic disease in Cuba]. / Caracterización fenotípica y factores de virulencia en cepas de Aeromonas aisladas de pacientes con enfermedad diarreica aguda en Cuba.

Fifty four strains of Aeromonas spp were isolated from patients with acute diarrheic episodes by using Aerokey II and Aeroesquema methods. In vitro antimicrobial susceptibility and virulence factors were analyzed. The most frequently isolated specie was Aeromonas caviae. Over 75% of strains exhibited resistance to penicillins and ce-phalosporins; for the other antibiotic groups resistance was under 20%. Twenty six strains (48.1 %) were multiresist-ant. At least one virulence factor among those evaluated in the study was present in 53 (98.1%) of the 54 strains.

Caracterización fenotípica y factores de virulencia en cepas de Aeromonas aisladas de pacientes con enfermedad diarreica aguda en Cuba / Phenotypic characteristics and virulence factors in Aeromonas strains isolated from patients with diarrheic disease in Cuba

Fifty four strains of Aeromonas spp were isolated from patients with acute diarrheic episodes by using Aerokey II and Aeroesquema methods. In vitro antimicrobial susceptibility and virulence factors were analyzed. The most frequently isolated specie was Aeromonas caviae. Over 75 percent of strains exhibited resistance to penicillins and ce-phalosporins; for the other antibiotic groups resistance was under 20 percent. Twenty six strains (48.1 percent) were multiresist-ant. At least one virulence factor among those evaluated in the study was present in 53 (98.1 percent) of the 54 strains.

Se identificaron 54 cepas de Aeromonas aisladas de pacientes con enfermedad diarreica aguda mediante los métodos Aerokey II y Aeroesquema. Se determinó la susceptibilidad antimicrobiana y algunos factores de virulencia. La especie encontrada en mayor frecuencia fue Aeromonas caviae. Se observaron valores de resistencia por sobre 75 por ciento para penicilinas y cefalotina; para el resto de los antimicrobianos estos valores se encontraron bajo 20 por ciento>; 26 cepas (48,1 por ciento) presentaron multi-resistencia. Al menos un factor de virulencia de los investigados estuvo presente en 53 (98,1 por ciento) de las 54 cepas analizadas.

Adapting to contour-clamped homogeneous electric field minichamber technology the PulseNet protocols to resolve XbaI-DNA fragments of Salmonella serotype Braenderup.

We propose cost-effective protocols for preparing and resolving the PulseNet universal DNA size standard in contour-clamped homogeneous electric field (CHEF) minichambers. Intact DNA molecules were prepared with protease-free solutions, and electrophoresis separations of the DNA standards needed 5.5h, giving band pattern resolutions similar to those attained with the PulseNet protocols standardized in CHEF chambers.

Caracterización microbiológica de cepas de Vibrio cholerae no-O1 aisladas en Cuba / Microbiological characterization of non-01 Vibrio cholerae isolated in Cuba

Se estudiaron 422 cepas de Vibrio cholerae no-O1 procedentes de 9 provincias del país, de ellas 9 aisladas de un brote de enfermedad de transmisión hídrica. En la totalidad de las cepas se determinó la susceptibilidad antimicrobiana y la presencia de factores de virulencia. En las 9 cepas procedentes del brote, se realizó además, el estudio de macrorrestricción de ADN mediante la técnica de electroforesis de campo pulsado. Se demostró por primera vez en Cuba y el Caribe, la circulación de cepas de V. cholerae no-O1 atípicas (resistentes al compuesto vibriostático O129 y al trimetoprim-sulfametoxazol). El comportamiento de la susceptibilidad antimicrobiana demostró por primera vez la circulación en Cuba de 2 patrones diferentes de resistencia (ampicilina, trimetoprim/sulfametoxazol, sulfonamida y tetraciclina, trimetoprim/sulfametoxazol, sulfonamida). La frecuencia de cepas resistentes al trimetoprim-sulfametoxazol se mantuvo similar en todo el período de estudio. Sin embargo, se produjo un decrecimiento de la resistencia a la ampicilina y un aumento de la resistencia a la tetraciclina. Los principales factores de virulencia que se encontraron fueron la gelatinasa, la hemolisina, la elastasa y la adherencia a células HEp-2. Sin embargo, las cepas del brote mostraron mayores porcentajes que el resto, para la presencia de la toxina termoestable y la presencia de fimbrias. Los resultados de los estudios molecular y epidemiológico permitieron dar una respuesta acelerada y precisa sobre la etiología del primer brote de enfermedad de transmisión alimentaria.

The study of 422 non-01 Vibrio cholerae strains from nine provinces, 9 of them isolated from a water-borne disease outbreak, was performed. All the strains exhibited antimicrobial susceptibility and virulence factors. The nine strains from the outbreak were subjected to a DNA macrorestriction study based on the pulsed field electrophoresis technique. For the first time in Cuba and the Caribbean, the circulation of atypical non-01 V. Cholerae strains (resistent to vibriostatic compound 0129 and trimethoprim/sulfamethoxazole). The behavior of antimicrobial susceptibility evinced for the first time the circulation of two different resistence patterns in Cuba (ampicilline, trimethoprim/sulfamethoxazole, sulfonamide and tetracycline, trimethoprim/sulfamethoxazole, sulfonamide). The frequency of trimethoprim/sulfamethoxazole-resistent strains was similar during the whole period of study. However, resistance to ampicilline decreased whereas resistance to tetracycline increased. The main found virulence factors were gelatinase, hemolysine, elastase and adherence to Hep-2 cells. On the other hand, the outbreak strains showed higher percentages than the others due to the presence of heat-liable toxin and fimbriae. The results of the molecular and epidemiological studies allowed giving a speedy and accurate response that explained the etiology of the first food-borne disease outbreak.

Caracterización microbiológica de cepas de Vibrio cholerae no-O1 aisladas en Cuba / Microbiological characterization of non-01 Vibrio cholerae isolated in Cuba

Se estudiaron 422 cepas de Vibrio cholerae no-O1 procedentes de 9 provincias del país, de ellas 9 aisladas de un brote de enfermedad de transmisión hídrica. En la totalidad de las cepas se determinó la susceptibilidad antimicrobiana y la presencia de factores de virulencia. En las 9 cepas procedentes del brote, se realizó además, el estudio de macrorrestricción de ADN mediante la técnica de electroforesis de campo pulsado. Se demostró por primera vez en Cuba y el Caribe, la circulación de cepas de V. cholerae no-O1 atípicas (resistentes al compuesto vibriostático O129 y al trimetoprim-sulfametoxazol). El comportamiento de la susceptibilidad antimicrobiana demostró por primera vez la circulación en Cuba de 2 patrones diferentes de resistencia (ampicilina, trimetoprim/sulfametoxazol, sulfonamida y tetraciclina, trimetoprim/sulfametoxazol, sulfonamida). La frecuencia de cepas resistentes al trimetoprim-sulfametoxazol se mantuvo similar en todo el período de estudio. Sin embargo, se produjo un decrecimiento de la resistencia a la ampicilina y un aumento de la resistencia a la tetraciclina. Los principales factores de virulencia que se encontraron fueron la gelatinasa, la hemolisina, la elastasa y la adherencia a células HEp-2. Sin embargo, las cepas del brote mostraron mayores porcentajes que el resto, para la presencia de la toxina termoestable y la presencia de fimbrias. Los resultados de los estudios molecular y epidemiológico permitieron dar una respuesta acelerada y precisa sobre la etiología del primer brote de enfermedad de transmisión alimentaria(AU)

The study of 422 non-01 Vibrio cholerae strains from nine provinces, 9 of them isolated from a water-borne disease outbreak, was performed. All the strains exhibited antimicrobial susceptibility and virulence factors. The nine strains from the outbreak were subjected to a DNA macrorestriction study based on the pulsed field electrophoresis technique. For the first time in Cuba and the Caribbean, the circulation of atypical non-01 V. Cholerae strains (resistent to vibriostatic compound 0129 and trimethoprim/sulfamethoxazole). The behavior of antimicrobial susceptibility evinced for the first time the circulation of two different resistence patterns in Cuba (ampicilline, trimethoprim/sulfamethoxazole, sulfonamide and tetracycline, trimethoprim/sulfamethoxazole, sulfonamide). The frequency of trimethoprim/sulfamethoxazole-resistent strains was similar during the whole period of study. However, resistance to ampicilline decreased whereas resistance to tetracycline increased. The main found virulence factors were gelatinase, hemolysine, elastase and adherence to Hep-2 cells. On the other hand, the outbreak strains showed higher percentages than the others due to the presence of heat-liable toxin and fimbriae. The results of the molecular and epidemiological studies allowed giving a speedy and accurate response that explained the etiology of the first food-borne disease outbreak(AU)

Estudio de marcadores fenotípicos y de susceptibilidad antimicrobiana en cepas de Escherichia coli entéricas / Study of phenotypical and antimicrobial susceptibility markers in enteric Escherichia coli strains

Se analizaron 41 cepas de Escherichia coli, aisladas en niños menores de 5 años con diarreas agudas, procedentes de las diferentes provincias del país. Fueron probados 4 importantes determinantes fenotípicos: sorbosa, sorbitol, enterohemolisina y la serología de O157: H7, para seleccionar las cepas de la categoría enterohemorrágica o productora de toxina shiga. De igual modo fueron caracterizadas por el método de biotipaje y de susceptibilidad antimicrobiana. El uso de las pruebas fenotípicas mostró 6 cepas con características presuntivas, de estas, 4 mostraron mayor probabilidad de pertenecer a la categoría productora de toxina shiga. En el ensayo de susceptibilidad antimicrobiana las cepas mostraron elevada resistencia, sobre todo para la ampicilina y el trimetropin-sulfametoxasol. Otro hallazgo de interés fueron los valores de resistencia y de susceptibilidad intermedia obtenidos para el augmentin, aztreonan y ceftriaxona. Se alcanzaron 12 patrones de resistencia antimicrobiana y de ellos 10 resaltaron por ser multirresistentes.

Forty strains of Escherichia coli isolated from children under 5 years of age with acute diarreas, coming from different provinces of the country , were analyzed. Four important phenotypical determinants were tested: sorbosa, sorbitol, enterohemolysin and 0157: H7 serology, in order to select those strains from enterohemorrhagic or Shiga toxin-producing category. Likewise, they were characterized by biotyping and antimicrobial susceptibility methods. The use of phenotypical tests showed six strains with presumptive characteristics, four of which were most likely to be Shiga toxin-producing strains. In antimicrobial susceptibility test, the strains showed high resistance mainly to ampicillin and trimethrophin-sulfamethoxasole. Another interesting finding were intermediate resistance and susceptibility values to augmentin, aztreonan and ceftriaxone. There were 12 antimicrobial resistance patterns of which 10 were multi-resistant.

Estudio de marcadores fenotípicos y de susceptibilidad antimicrobiana en cepas de Escherichia coli entéricas / Study of phenotypical and antimicrobial susceptibility markers in enteric Escherichia coli strains

Se analizaron 41 cepas de Escherichia coli, aisladas en niños menores de 5 años con diarreas agudas, procedentes de las diferentes provincias del país. Fueron probados 4 importantes determinantes fenotípicos: sorbosa, sorbitol, enterohemolisina y la serología de O157: H7, para seleccionar las cepas de la categoría enterohemorrágica o productora de toxina shiga. De igual modo fueron caracterizadas por el método de biotipaje y de susceptibilidad antimicrobiana. El uso de las pruebas fenotípicas mostró 6 cepas con características presuntivas, de estas, 4 mostraron mayor probabilidad de pertenecer a la categoría productora de toxina shiga. En el ensayo de susceptibilidad antimicrobiana las cepas mostraron elevada resistencia, sobre todo para la ampicilina y el trimetropin-sulfametoxasol. Otro hallazgo de interés fueron los valores de resistencia y de susceptibilidad intermedia obtenidos para el augmentin, aztreonan y ceftriaxona. Se alcanzaron 12 patrones de resistencia antimicrobiana y de ellos 10 resaltaron por ser multirresistentes(AU)

Forty strains of Escherichia coli isolated from children under 5 years of age with acute diarreas, coming from different provinces of the country , were analyzed. Four important phenotypical determinants were tested: sorbosa, sorbitol, enterohemolysin and 0157: H7 serology, in order to select those strains from enterohemorrhagic or Shiga toxin-producing category. Likewise, they were characterized by biotyping and antimicrobial susceptibility methods. The use of phenotypical tests showed six strains with presumptive characteristics, four of which were most likely to be Shiga toxin-producing strains. In antimicrobial susceptibility test, the strains showed high resistance mainly to ampicillin and trimethrophin-sulfamethoxasole. Another interesting finding were intermediate resistance and susceptibility values to augmentin, aztreonan and ceftriaxone. There were 12 antimicrobial resistance patterns of which 10 were multi-resistant(AU)

[Study of phenotypical and antimicrobial susceptibility markers in enteric Escherichia coli strains]. / Estudio de marcadores fenotípicos y de susceptibilidad antimicrobiana en cepas de Escherichia coli entéricas.

Forty strains of Escherichia coli isolated from children under 5 years of age with acute diarreas, coming from different provinces of the country , were analyzed. Four important phenotypical determinants were tested: sorbosa, sorbitol, enterohemolysin and 0157:H7 serology, in order to select those strains from enterohemorrhagic or Shiga toxin-producing category. Likewise, they were characterized by biotyping and antimicrobial susceptibility methods. The use of phenotypical tests showed six strains with presumptive characteristics, four of which were most likely to be Shiga toxin-producing strains. In antimicrobial susceptibility test, the strains showed high resistance mainly to ampicillin and trimethrophin-sulfamethoxasole. Another interesting finding were intermediate resistance and susceptibility values to augmentin, aztreonan and ceftriaxone. There were 12 antimicrobial resistance patterns of which 10 were multi-resistant.

[Microbiological characterization of non-O1 Vibrio cholerae isolated in Cuba]. / Caracterización microbiológica de cepas de Vibrio cholerae no-O1 aisladas en Cuba.

The study of 422 non-01 Vibrio cholerae strains from nine provinces, 9 of them isolated from a water-borne disease outbreak, was performed. All the strains exhibited antimicrobial susceptibility and virulence factors. The nine strains from the outbreak were subjected to a DNA macrorestriction study based on the pulsed field electrophoresis technique. For the first time in Cuba and the Caribbean. The circulation of atypical non-01 V cholerae strains (resistent to vibriostatic compound 0129 and trimethoprim/sulfamethoxazole). The behavior of antimicrobial susceptibility evinced for the first time the circulation of two different resistence patterns in Cuba (ampicilline, trimethoprim/ sulfamethoxazole, sulfonamide and tetracycline, trimethoprim/ sulfamethoxazole, sulfonamide). The frequency of trimethoprim/ sulfamethoxazole-resistent strains was similar during the whole period of study. However, resistance to ampicilline decreased whereas resistance to tetracycline increased. The main found virulence factors were gelatinase, hemolysine, elastase and adherence to Hep-2 cells. On the other hand, the outbreak strains showed higher percentages than the others due to the presence of heat-liable toxin and fimbriae. The results of the molecular and epidemiological studies allowed giving a speedy and accurate response that explained the etiology of the first food-borne disease outbreak.

Estudio de correlación entre cepas Escherichia coli sorbosa negativa y su capacidad de producir enterotoxinas / Study of correlation between negative sorbose E. coli strains and their enterotoxin-producing capacity

Se estudiaron 50 cepas de Escherichia coli sorbosa negativas, aisladas de niños con diarreas líquidas de aparición espontánea. La identificación hasta especie se realizó por métodos convencionales, incluidos los marcadores fenotípicos sorbosa y sorbitol. Las técnicas de reacción en cadena de la polimerasa e hibridación de ADN, fueron realizadas para confirmar la capacidad de producción de enterotoxinas. Se obtuvo muy buena correlación entre las cepas de Escherichia coli sorbosa negativas y su capacidad de elaborar enterotoxinas termolábil y termoestable. El estudio demostró un alto porcentaje de estas cepas con fragmentos de genes codificadores, 94 y 44 % respectivamente, predominando la enterotoxina termoestable. Se obtuvieron los principales biotipos primarios de importancia taxonómica y epidemiológica. Se demostró que el método de la sorbosa posee cualidades que lo distinguen como marcador fenotípico, con 100 % de sensibilidad y 88 % de especificidad, revelándose como método de diagnóstico presuntivo práctico y de gran utilidad para los Laboratorios de Microbiología Clínica.

Fifty negative sorbose Escherichia coli strains isolated from children who suffered spontaneously occurring fluid diarrheas were studied. Identification up to the level of species was performed by conventional methods including phenotypical markers known as sorbose and sorbitol. The PCR and DNA gene hybridization were applied to confirm the enterotoxin-producing capacity. A good correlation between negative sorbose Escherichia coli and their capacity of producing heat-labile and heat -stable enterotoxin was observed. The study showed a high percentage of these strains with encoding gene fragments, 94 and 44% respectively, being heat-stable enterotoxin the predominant. The main primary biotypes of taxonomic and epidemiological importance were obtained. It was demonstrated that the sorbose method has certain qualities that make it stand out as a phenotypical marker, with 100% sensitivity and 88% specificity, and as a practical presumptive diagnostic method that is very useful for Clinical Microbiology Laboratories.

Estudio de correlación entre cepas Escherichia coli sorbosa negativa y su capacidad de producir enterotoxinas / Study of correlation between negative sorbose E. coli strains and their enterotoxin-producing capacity

Se estudiaron 50 cepas de Escherichia coli sorbosa negativas, aisladas de niños con diarreas líquidas de aparición espontánea. La identificación hasta especie se realizó por métodos convencionales, incluidos los marcadores fenotípicos sorbosa y sorbitol. Las técnicas de reacción en cadena de la polimerasa e hibridación de ADN, fueron realizadas para confirmar la capacidad de producción de enterotoxinas. Se obtuvo muy buena correlación entre las cepas de Escherichia coli sorbosa negativas y su capacidad de elaborar enterotoxinas termolábil y termoestable. El estudio demostró un alto porcentaje de estas cepas con fragmentos de genes codificadores, 94 y 44 por ciento respectivamente, predominando la enterotoxina termoestable. Se obtuvieron los principales biotipos primarios de importancia taxonómica y epidemiológica. Se demostró que el método de la sorbosa posee cualidades que lo distinguen como marcador fenotípico, con 100 por ciento de sensibilidad y 88 por ciento de especificidad, revelándose como método de diagnóstico presuntivo práctico y de gran utilidad para los Laboratorios de Microbiología Clínica(AU)