The causative agent of cholera, Vibrio cholerae, can enter into a viable but non-culturable (VBNC) state in response to unfavorable conditions. The aim of this study was to evaluate the in situ survival of V. cholerae in an aquatic environment of the Southern Caribbean Sea, and its induction and resuscitation from the VBNC state. V. cholerae non-O1, non-O139 was inoculated into diffusion chambers placed at the Cuare Wildlife Refuge, Venezuela, and monitored for plate, total and viable cells counts. At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state. Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells. Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season. Resuscitation studies showed significant recovery of cell cultivability with spent media addition (p < 0.05). These results suggest that V. cholerae can persist in the VBNC state in this Caribbean environment and revert to a cultivable form under favorable conditions. The VBNC state might represent a critical step in cholera transmission in susceptible areas.

El agente causal del cólera, Vibrio cholerae, puede entrar a un estado viable no cultivable (VNC) en respuesta a condiciones desfavorables. El objetivo de este estudio fue evaluar la supervivencia in situ de V. cholerae en un ambiente acuático al sur del Mar Caribe y su inducción y resucitación del estado VBNC. V. cholerae no-O1, no-O139 fue inoculado en cámaras de difusión ubicadas en el Refugio de Fauna Cuare, Venezuela, y monitoreado para contaje de colonias, células totales y viables. En 119 días de exposición al ambiente, el contaje de colonias fue < 10 UFC/mL y una fracción de la población bacteriana entró al estado VBNC. Adicionalmente, la viabilidad disminuyó dos órdenes de magnitud y ocurrieron cambios morfológicos de células bacilares a cocoides. Entre las variables del ambiente acuático, la salinidad presentó correlación negativa con el contaje de colonias. Los estudios de resucitación mostraron recuperación significativa de la cultivabilidad celular con adición de sobrenadantes de cultivos en crecimiento activo (p < 0.05). Estos resultados sugieren que V. cholerae puede persistir en estado VBNC en este ambiente de Caribe y revertir a una forma cultivable bajo condiciones favorables. El estado VBNC podría representar un paso crítico en la transmisión del cólera en áreas susceptibles.

The first report to our knowledge, of hyperinfection by Strongyloides stercoralis (HS) and hypereosinophilia, associated to immune suppression by Rituximab (the only drug received for the last one year and 10 months), in a patient with mantle-cell lymphoma (MCL), is presented. The patient has a 3-year history of MCL, and developed two accesses of HS during 2008, including meningitis, pneumonia and presence of larvae of S. stercoralis in the lungs. We had a unique chance to look at cytotoxicity of filariform larvae in the expectoration after Ivermectin treatment, showing immobilization and death of larvae, associated with eosinophils attached to the cuticle of the parasite.

Se presenta el primer reporte, hasta donde tengamos información, de hiperinfección por Strongyloides stercoralis (HS) e hipereosinofilia asociados a inmunosupresión por Rituximab (el único medicamento recibido durante 1 año y 10 meses), en un paciente con linfoma de células del manto (LCM). La paciente tuvo una historia de 3 años con LCM, y desarrolló 2 accesos de HS durante el 2008, incluyendo meningitis, neumonía y presencia de larvas de S. stercoralis en los pulmones. Se tuvo la oportunidad única de observar la citotoxicidad contra las larvas filariformes en la expectoración, luego del tratamiento con Ivermectina, mostrando la inmovilización y muerte de las larvas, asociada a la presencia de eosinófilos adheridos a la cutícula del parásito.

Caso de meningite por Staphylococcus warneri em paciente com hiperinfecção com Strongyloides stercoralis, possivelmente associada com tratamento de rituximab para linfoma de células do manto é relatada pela primeira vez na literatura. A paciente, mulher de 59 anos com história de três anos de linfoma aparentemente bem controlado com tratamento com quimioterapia-imunoterapia e, em seguida, somente imunoterapia e diagnóstico de estrongiloidíase. Meningite foi diagnosticada por cultura do líquido cefalorraquidiano e testada com sistema automatizado de placa. A paciente foi tratada com sucesso com vancomicina, embora a febre e a tosse produtiva não tenham desaparecido. Após graves sintomas gastrointestinais a paciente desenvolveu pneumonia três semanas mais tarde. Síndrome de hiperinfecção por S. stercoralis foi diagnosticada, com larvas abundantes nas fezes e expectoração.

A case of meningitis due to Staphylococcus warneri in a patient with a hyperinfection with Strongyloides stercoralis possibly associated with rituximab treatment for mantel cell lymphoma is reported for the first time in the literature. The patient was a 59-year-old woman, with a 3-year history of an apparently well controlled lymphoma after treatment with chemotherapy-immunotherapy and then immunotherapy alone, and diagnosis of strongyloidiasis. Meningitis was diagnosed by cerebrospinal fluid culture and tested with an automated plate system. The patient was successfully treated with vancomycin; although fever and productive cough persisted. Severe gastrointestinal symptoms and pneumonia developed three weeks later. Hyperinfection syndrome by S. stercoralis was diagnosed, with abundant larvae in feces and expectoration.

A proven case of human infection caused by Angiostrongylus costaricensis is reported for the first time in Venezuela. The patient was a 57-year-old female surgically operated because of signs of peritonitis with a palpable mass at the lower right quadrant of the abdomen. WBC count reported 16,600 cells/mm³, with 46% eosinophils. The tumoral aspect of ileocolic area and peritoneal lymph nodes prompted the resection of a large area of the terminal ileum, cecum, part of the ascending colon and a small part of the jejunum, where a small lesion was found. The pathology showed thickened areas of the intestinal wall with areas of hemorrhage and a perforation of the cecum. Histology showed intense eosinophil infiltration of the whole intestinal wall, granulomas with giant cells and eosinophils. Some of the granuloma surrounded round or oval eggs with content characterized by a large empty area, cells or embryo in the center, and sometimes nematode larvae. A cross section of an adult nematode worm was observed inside a branch of mesenteric artery. The intestinal affected area, the characteristics of the lesions, the presence of eggs in the submucosa with nematode larvae inside, and the observation of a nematode inside a mesenteric artery, makes sufficient criteria for the diagnosis of an infection by Angiostrongylus costaricensis.

Un caso comprobado de infección humana por Angiostrongylus costaricensis es reportado para la literatura por vez primera en Venezuela. El paciente fue una mujer de 57 años de edad intervenida quirúrgicamente debido a signos de peritonitis, con una masa palpable en la fosa ilíaca derecha del abdomen. El recuento de leucocitos reportó 16.600 células/mm³ con 46% eosinófilos. El aspecto tumoral del área íleocólica y las adenopatías peritoneales encontradas motivó la resección de un área grande del ileon terminal, ciego, parte del colon ascendente y una pequeña parte del yeyuno donde se encontró una pequeña lesión interpretada como metastásica. La patología mostró áreas engrosadas de la pared intestinal con zonas de hemorragia y perforación del ciego. El estudio histológico mostró una intensa infiltración eosinofílica de toda la pared intestinal, granulomas con células gigantes y eosinófilos. Algunos de los granulomas rodeaban huevos ovales o redondos con un contenido caracterizado por una gran área vacía, células o embriones en el centro, y en algunos casos una larva de nematodo. Un corte transversal de un verme nematodo adulto se observó dentro de una arteria mesentérica. El área intestinal afectada, las características de las lesiones, la presencia de huevos en la submucosa con larvas de nematodo dentro, y la observación de un nematodo dentro de una arteria mesentérica, aportan suficientes criterios para el diagnóstico de una infección por Angiostrongylus costaricensis.

Fasciolíase é uma doença emergente/re-emergente transmitida por vetores com a distribuição sabidamente mais ampla. Existem aproximadamente 17 milhões de pessoas infectadas em todo mundo, sendo a região andina a área mais afetada. Há uma necessidade importante para desenvolver ferramentas diagnósticas sensíveis e específicas para tratar cedo os pacientes e para evitar complicações. Neste trabalho avaliamos a resposta imune de seres humanos infectados comparando a duas preparações antigênicas: o extrato solúvel total (FhTSE) e o vômito (FhAWV) do verme adulto a fim de identificar as frações antigênicas específicas para Fasciola hepatica. Ambas as preparações foram processadas por SDS-PAGE e Western blot com os soros humanos de portadores de fasciolíase (F), outras parasitoses e indivíduos saudáveis. No immunoblot de FhTSE, os soros F reconheceram 16 faixas com PM entre 8 e 110 kDa, das quais as de 8, 9, 10, 38, 45 e 57 kDa foram específicas. Na preparação de FhAWV, os soros F reconheceram 9 faixas com PM entre 8 e 85 kDa, das quais as de 8, 12, 15 e 24 kDa foram específicas. Algumas faixas com reação cruzada foram evidentes com os soros dos pacientes com outras parasitoses, mais freqüentes com o FhTSE. As faixas dentro do PM mencionado, particularmente aquela de 8 kDa, mostraram ser específicas por outros autores, e merecem a caracterização adicional para seu uso potencial no diagnóstico imunológico.

Fascioliasis is an emerging/re-emerging vector-borne disease with the widest known distribution. Approximately 17 million people are infected around the world, being the Andean region the most affected area. There is an important necessity to develop sensitive and specific diagnostic tools to treat patients early and to avoid complications. In this paper we evaluated the immune response of infected humans against two antigenic preparations: the total soluble extract (FhTSE) and the adult worm vomit (FhAWV) in order to identify antigenic fractions specific for Fasciola hepatica. Both preparations were processed by SDS-PAGE and Western blot with human sera with fascioliasis (F), other parasitosis and healthy individuals. In the immunoblot of FhTSE, sera F recognised 16 bands with MW between eight and 110 kDa, from which those of 8, 9, 10, 38, 45 and 57 kDa were specific. In the preparation FhAWV, sera F recognised nine bands with MW from eight to 85 kDa, from which those of 8, 12, 15 and 24 kDa were specific. Some bands of cross-reaction were evident with sera from patients with other parasitoses, more frequent with the FhTSE. Bands within the MW mentioned, particularly that of eight kDa, have been shown to be specific by others, and deserve additional characterisation for their potential use in immunodiagnosis.

Resumen. Se reporta para la literatura parasitológica venezolana, un nuevo caso de infección humana por Fasciola hepatica. Se trata de una paciente de 81 años, asintomática, quien presentó durante exámenes de rutina, una eosinofilia de 21% y una eritrosedimentación de 26 mm/h. Estos valores fueron normales para el momento del tratamiento (4 meses después), así como también lo eran varias pruebas de funcionamiento hepático, química sanguínea y valores hematológicos. El número de huevos del parásito fue de 90 a 130/g de heces. La paciente fue exitosamente tratada con la formulación humana de triclabendazol en 2 dosis simples de 10 mg/kg cada una separadas por 24h, resultando negativas las evaluaciones coproparasitológicas realizadas a los 12, 18, 60 y 120 días post-tratamiento. Los mismos tests de laboratorio arriba mencionados no mostraron modificaciones excepto por un incremento leve en el contaje de eosinófilos en los días 2 y 11, así como un incremento en los valores de eritrosedimentación al día 11 post-tratamiento. Así mismo, no se registraron síntomas durante 2 meses de seguimiento post-tratamiento. La paciente probablemente adquirió la infección en su hogar, a través de la ingestión de lechuga adquirida en el comercio local, originalmente proveniente de un área endémica de F. hepatica bovina distante de su hogar, entre 4 y 7 meses previos a su tratamiento médico. Enfatizamos la necesidad de investigar esta parasitosis, la cual ha sido subvalorada como infección humana en Venezuela, probablemente debido al limitado conocimiento que de ella tiene el personal de salud, su sintomatología poco específica y la ausencia en el país de procedimientos diagnósticos más sensibles.

Abstract. A new case of human infection due to Fasciola hepatica is reported in the Venezuelan parasitological literature. The patient is an 81-year-old female asymptomatic, with an eosinophilia of 21% and eritrosedimentation rate of 26 mm/h and was found during a routine check up. These values were normal at the time of treatment (4 months later), as were several tests of hepatic function, blood chemistry and peripheral blood haematological values. The number of eggs of the parasite were between 90 and 130/g of faeces. The patient was successfully treated with the human formulation of triclabendazole at 2 single doses of 10mg/kg, each separated by 24h. The same laboratory tests mentioned above did not show modifications, except for a slight increase in the eosinophil counts at 2 and 11 days and erythrosedimentation rate at 11 days post-treatment. Coproparasitological observations carried out at 12, 18, 60 and 120 days post-treatment were negative. No clinical symptoms were registered up to 2 months after treatment. The patient most probably acquired the infection at home through the ingestion of commercially available lettuce from an endemic area of bovine fascioliasis, very distant from home, and between 4 and 7 months previous to treatment. We stress the need to investigate F. hepatica infections which he been neglected, probably because of limited knowledge by health workers, lack of specific symptoms and absence of more sensitive diagnostic procedures.