ABSTRACT Introduction: Cholera is an acute intestinal infection caused by toxigenic strains of Vibrio choleare. The rapid dissemination and emergence of the multiresistance that characterizes this pathogen could interfere with the success of antimicrobial therapy, so it is a priority to monitor changes in susceptibility patterns, as a transcendental part of the resistance control policy antimicrobial. Objective: To determine the behavior of antimicrobial resistance against the antimicrobials of interest used in the treatment, the presence of enzymatic virulence factors and whether there is a relationship between them. Methods: A descriptive cross-sectional study was conducted during July 2012 to December 2015. Where 500 isolates belonging to the cepary of the National Reference Laboratory for Acute Diarrheal Diseases of the Institute of Tropical Medicine "Pedro Kourí", from outbreaks of EDA of the national network of Microbiology laboratories in Cuba. Conventional phenotypic methods were applied to determine the behavior of antimicrobial resistance, the presence of enzymatic factors and their relationship with antimicrobial resistance. Results: The highest percentages of sensitivity were obtained against azithromycin (98%), doxycycline (96%) and ciprofloxacin (93%) and resistance to ampicillin (100%) and trimethoprim-sulfamethoxazole (99.4%). 44 isolated (8.8%) multi-resistant were found. All isolates had at least two extracellular enzymes as virulence factors, the most frequent: gelatinase (96%) and lecithinase (95%). Conclusions: There is a direct and proportional relationship between the presence of virulence factors and antimicrobial resistance, synergism that arises greater pathogenicity of the isolates studied from epidemic outbreaks.

RESUMEN Introducción: El cólera es una infección intestinal aguda causada por cepas toxigénicas de Vibrio choleare. La rápida diseminación y emergencia de la multirresistencia que caracteriza a este patógeno, podría interferir en el éxito de la terapia antimicrobiana, por lo que constituye una prioridad monitorear los cambios en los patrones de susceptibilidad, como parte trascendental de la política de control de la resistencia antimicrobiana. Objetivo: Determinar el comportamiento de la resistencia antimicrobiana frente a los antimicrobianos de interés empleados en el tratamiento, la presencia de factores de virulencia enzimáticos y si existe relación entre ambos. Métodos: Se realizó un estudio descriptivo de corte transversal durante julio de 2012 a diciembre de 2015. Se estudiaron 500 aislamientos pertenecientes al cepario del Laboratorio Nacional de Referencia de Enfermedades Diarreicas Agudas del Instituto de Medicina Tropical “Pedro Kourí”, procedentes de brotes de enfermedades diarreicas agudas de la red nacional de laboratorios de Microbiología de Cuba. Se aplicaron métodos convencionales fenotípicos para determinar el comportamiento de la resistencia antimicrobiana, la presencia de factores enzimáticos y la relación de estos con la resistencia antimicrobiana. Resultados: Los mayores porcentajes de sensibilidad se obtuvieron frente a azitromicina (98 %), doxiciclina (96 %) y ciprofloxacina (93%) y de resistencia frente a ampicilina (100 %) y trimetoprim-sulfametoxazol (99,4 %). Se encontraron 44 aislados (8,8 %) multirresistente. Todos los aislamientos poseían al menos dos enzimas extracelulares como factores de virulencia, las más frecuentes: gelatinasa (96 %) y lecitinasa (95 %). Conclusiones: Se evidencia una relación directa y proporcional entre la presencia de los factores de virulencia y resistencia antimicrobiana, sinergismo que surgiere mayor patogenicidad de los aislados estudiados procedentes de brotes epidémicos.

ABSTRACT A brief summary is made on the evolution of health coverage in the mountainous area of the province of Cienfuegos. It takes into account the complaint made by the Commander-in-Chief in the trial for the events of the Moncada barracks and his plea "History will absolve me" where the precarious situation of health in our fields is addressed and the solutions given from the establishment of the Rural Medical Service Program, the integration of Healthcare Teaching through Healthcare, the Piti Fajardo Contingent (6th year) and the creation by Fidel himself of the Family Medicine Program and the specialty of Comprehensive General Medicine, with which achieves health coverage. The graduation of the first specialists in Comprehensive General Medicine and Comprehensive General Stomatology trained entirely in the Clinics of the mountainous massif of our Cienfuegos Turquino Plan is noted as a colophon.

RESUMEN Se hace un breve resumen sobre la evolución de la cobertura salubrista en la zona montañosa de la provincia de Cienfuegos. Se tiene en cuenta la denuncia realizada por el Comandante en Jefe en el juicio por los hechos del cuartel Moncada y su alegato ¨La Historia me Absolverᨠdonde se aborda la situación precaria de la salud en nuestros campos y las soluciones dadas a partir del establecimiento del Programa del Servicio Médico Rural, la integración Docente Asistencial a través de los Asistenciales, del Contingente Piti Fajardo (6to año) y la creación por el propio Fidel del Programa de la Medicina Familiar y la especialidad de Medicina General Integral, con lo cual se logra la cobertura en salud. Se señala como colofón la graduación de los primeros especialistas en Medicina General Integral y Estomatología General Integral formados íntegramente en los Consultorios del macizo montañoso de nuestro Plan Turquino cienfueguero.

ABSTRACT Vibrio cholerae non-O1, non-O139 is the third bacterium group from the genus Vibrio most commonly causing diarrhea. It survives in aquatic environments, using the formation of biofilm as a survival mechanism facilitating the transmission of diarrheal disease. Multi-drug resistant V. cholerae isolates have been characterized since the year 1977, and some of the mechanisms involved include the production of extended-spectrum β-lactamases (ESBLs). The purpose of the study was to determine the formation of biofilm in Cuban isolates of V. cholerae non-O1, non-O139 causing acute diarrheal disease (ADD), and detect the production of ESBLs in those with total or intermediate resistance to ampicillin. A descriptive cross-sectional study was conducted from January 2014 to June 2015. The study sample was 55 previously characterized isolates obtained from the strain collection at the ADD National Reference Laboratory of Pedro Kourí Institute. For phenotypic determination of ESBLs, 43 were studied which were known to be susceptible to ampicillin. 54.5 % of the isolates tested positive for biofilm formation, with a predominance of those classified as moderate (46.6 %) and weak (36.6 %) biofilm producers. Of the 34 isolates resistant to ampicillin, 26.5 % were positive for ESBL production. Of the 9 with intermediate ampicillin resistance, 44.4 % were positive. The results of the present study contribute knowledge about their ability to persist in the environment, and provide insight into antimicrobial resistance mechanisms.

RESUMEN Vibrio cholerae no-O1, no-O139 es el tercer grupo de bacterias del género Vibrio que con más frecuencia producen diarreas. Sobrevive en los ambientes acuáticos, utilizando la formación de biopelícula como mecanismo de supervivencia que propicia la transmisión de la enfermedad diarreica. Desde 1977 se caracterizan aislados de V. cholerae con resistencia múltiple, y algunos de los mecanismos involucrados incluyen la producción de β-lactamasas de espectro extendido (BLEE). Este trabajo tuvo como objetivo determinar la formación de biopelícula en los aislados cubanos de V. cholerae no-O1, no-O139, causantes de enfermedad diarreica aguda (EDA), y detectar la producción de BLEE en aquellos con resistencia total e intermedia a ampicilina. Se realizó un estudio descriptivo de corte transversal, entre enero 2014 y junio 2015. Se estudiaron 55 aislados caracterizados previamente, que formaban parte del cepario del Laboratorio Nacional de Referencia de EDA del Instituto “Pedro Kourí”. Para la determinación fenotípica de BLEE se estudiaron 43, de los que ya se conocía su susceptibilidad a ampicilina. El 54,5 % de los aislados resultaron positivos a la formación de biopelícula, y predominaron los clasificados como formadores moderados (46,6 %) y débiles (36,6 %). De los 34 resistentes a ampicilina, 26,5 % resultaron positivos a la producción de BLEE. En el caso de los nueve aislados con resistencia intermedia a ampicilina, 44,4 % resultaron positivos. Los resultados del presente estudio contribuyen al conocimiento sobre la capacidad que tienen de persistir en el ambiente y permiten profundizar sobre los mecanismos de resistencia a los antimicrobianos.

ABSTRACT INTRODUCTION In the Caribbean region, acute diarrheal diseases caused by the Vibrio genus have increased in recent years, following the 2010 earthquake in Haiti. Based on its capsular lipopolysaccharide, Vibrio cholerae is classified into more than 200 serogroups, divided into O1, O139 and non-O1, non-O139. Non-O1 serogroups produce clinical features ranging from mild diarrhea to severe dehydration. In Cuba, circulation of non-O1, non-O139 V. cholerae has been reported both in outbreaks and sporadic cases. OBJECTIVE Describe the antimicrobial susceptibility of V. cholerae to the drugs of interest used in its treatment and verify the presence of enzymatic virulence factors. METHODS A descriptive, cross-sectional study was conducted in January through November 2014, based on 125 non-O1, non-O139 V. cholerae isolates obtained during 2013 and 2014 from patients with acute diarrheal disease (isolates from the National Reference Laboratory for Acute Diarrheal Diseases of the Pedro Kouri Tropical Medicine Institute, Havana). Bacteriological identification was performed according to conventional methods. Antimicrobial susceptibility was determined by the Bauer-Kirby agar diffusion method. The technique described by Robinson in 1986 was used to determine virulence factors, hemolytic activity and enzyme factors (DNase, elastase, gelatinase), and Karagozova’s method for the enzyme lecithinase. RESULTS Highest percentages of sensitivity were obtained for azithromycin (98.4%), doxycycline (97.6%) and ciprofloxacin (96.8%), and highest resistance values for ampicillin (60%), sulfonamide (46.4%) and trimethoprim–sulfamethoxazole (32%). Six resistance patterns were detected, four found in Cuba for the first time, as well as six patterns of multidrug resistance (4.8%). All isolates had at least two extracellular enzymes as virulence factors. The most frequent were gelatinase (90.4%) and lecithinase (77.6%). The percentage of virulence factors was lower in the group of isolates resistant to ≥2 antimicrobials. An inverse relationship was found between presence of enzymatic virulence factors and resistance in the isolates studied. CONCLUSIONS The results suggest that azithromycin, doxycycline and ciprofloxacin should continue to be used for treatment of V. cholerae-caused infections, and confirmed the presence of four new resistance patterns in isolates circulating in Cuba.

Parasitologists from different medical institutions in Havana City carried out the external validation of ENZYMEBA, a diagnostic procedure of intestinal amebiasis developed at the "Pedro Kourí" Institute of Tropical Medicine. To this end, serial faeces specimens from 212 individuals were collected and observed on the microscope (reference test). The ENZYMEBA immunoassay (validation test) was also made. On comparing ENZYMEBA with the microscopic examination, satisfactory indexes of sensitivity and specificity were found. No cross-reactions were detected in faeces specimens, where found other parasites were present, too. Taking into account that only one faeces specimen per patient is enough for the diagnosis of intestinal amebiasis with ENZYMEBA, this procedure may be useful in studies of therapeutical efficacy and prevalence.

Parasitólogos de diferentes instituciones médicas de Ciudad de La Habana realizaron la validación externa de ENZYMEBA, procedimiento diagnóstico de amebiosis intestinal desarrollado en el Instituto de Medicina Tropical "Pedro Kourí". Para ello, fueron colectadas muestras seriadas de heces de 212 individuos sobre las que se realizó la observación microscópica (prueba de referencia) y el inmunoensayo ENZYMEBA (prueba en validación). ENZYMEBA, comparada con el examen microscópico de heces, mostró satisfactorios índices de sensibilidad y especificidad. No se observaron reacciones cruzadas con muestras de heces en que estaban presentes otros parásitos. Además, si se tiene en cuenta que para el diagnóstico de amebiosis intestinal con ENZYMEBA es suficiente una sola muestra de heces por paciente, este procedimiento pudiera ser útil en estudios de eficacia terapéutica y de prevalencia.