GroEL PCR- RFLP - An efficient tool to discriminate closely related pathogenic Vibrio species.

Vibrio sp. are autochthonous to marine and estuarine waters. Several species of Vibrio are pathogens. It is of utmost importance to detect and discriminate the Vibrio sp. that are often involved in food and water borne infections. Since 16S rRNA based identification has limited utility in differentiating the closely related pathogenic species from non pathogenic species, we have evaluated the discriminatory power of groEL PCR-RFLP for identification of closely related Vibrio sp. Accordingly, in the current study, the efficiency of groEL PCR- RFLP for detection and accurate differentiation of known pathogens among Vibrio sp. such as V. cholerae, V. parahaemolyticus, V. vulnificus, V. mimicus, V. fluvialis, V. alginolyticus, V. anguillarum was evaluated. PCR amplified groEL gene fragment of each Vibrio sp. was digested separately using 5 restriction enzymes viz. Hha1, Rsa1, Alu1, Dde1 and Mbo1. The accuracy of the method was further validated by insilico restriction analysis of multiple strains of each species using NEBcutter. The method proved to be efficient for detection and differentiation of Vibrio species under study. Phylogenetic analysis also revealed groEL gene to be a better phylogenetic marker for Vibrio compared to 16S rRNA. Hence, the method can be employed for accurate detection of Vibrio sp. including the closely related species.

Characterization of blooming algae and bloom-associated changes in the water quality parameters of traditional pokkali cum prawn fields along the South West coast of India.

In tropical shrimp farms, especially in traditional pokkali shrimp ponds, poor water quality management can lead to serious threats like heavy algal blooms and frequent disease outbreaks. An investigation regarding the frequent disease outbreaks in selected pokkali shrimp pond adjoining the Cochin backwaters has been conducted. Water quality parameters were analyzed during the study period. Results indicate a considerable fluctuation in the concentration of dissolved oxygen (DO) in the study in the shrimp farm where the algal bloom was observed. Poor water exchange along with nutrient loading from adjacent housing areas resulted in heavy algal bloom in the pond which led to hypoxic conditions in early morning and supersaturation of DO in the afternoon. It also led to considerably high alkaline pH. High levels of total ammonia nitrogen (TAN) were recorded in the sampling sites. Heavy algal bloom was observed throughout the study period in the selected pond especially in the summer. Anabaenopsis elenkinii and Merismopedia elagans which were the dominant species from the culture pond caused the bloom. Characterization and percentage distribution of bacteria present in the water and those associated with the algal blooms were carried out. The algae were found to support greater diversity of bacteria when compared to water. Pathogenic species like Aeromonas hydrophila and Vibrio parahaemolyticus were encountered in the study. Experimental challenge studies using Artemia as a model showed that the V. parahaemolyticus isolates were highly pathogenic. Hence, this study reveals how algal growth supports opportunistic pathogens in great diverse in a shrimp pond and causes frequent disease outbreaks under favourable conditions.

Comparison of cyclosporine A with methylprednisolone for treatment of canine atopic dermatitis: a parallel, blinded, randomized controlled trial.

The objective of this multicentre, parallel, blinded, randomized controlled study was to evaluate the efficacy and the safety of cyclosporine (CsA group, 117 dogs) in comparison with methylprednisolone (MP group, 59 dogs) in the treatment of atopic dermatitis for 4 months. Mean induction dose of both drugs (5 mg/kg CsA, 0.75 mg/kg MP) was tapered over time according to the clinical response. At the end of the study, the mean estimated percentage reduction from baseline (confidence interval) of lesion scores was 52% (44-59) and 45% (35-56), and the reduction in pruritus score was 36% (27-43) and 33% (23-43) in dogs in the CsA and MP groups, respectively. These percentages were not significantly different between groups. A significantly better overall assessment of efficacy was obtained in the CsA-treated dogs (76 vs. 63% responses excellent or good in the CsA compared with MP group). CsA-treated dogs presented a higher frequency of gastrointestinal disorders, mainly vomiting, but MP dogs tended to be more susceptible to infections. There was no remarkable change over baseline of the haematological and biochemical parameters in the two groups.

Résumé Le but de cette étude contrôlée, multicentrique, parallèle, en aveugle et randomisée, était d'évaluer l'efficacité et l'inocuité de la cyclosporine (groupe CsA, 117 chiens) en comparaison de la methylprednisolone (groupe MP, 59 chiens) pour le traitement de la dermatite atopique pendant 4 mois. La dose moyenne d'induction pour les deux molécules (5 mg kg−1 CsA, 0.75 mg kg−1 MP) était progressivement diminuée en fonction de la réponse clinique observée. A la fin de l'étude, le pourcentage moyen de réduction des scores lésionnels et les intervalles de confiance étaient de 52% (44-59) pour le groupe CsA et de 45% (35-56) pour le groupe MP, alors que le pourcentage moyen de réduction du prurit était de 36 % (27-43) pour la CsA et de 33% (23-43) pour la MP. Ces pourcentages ne sont pas significativement différents entre les groupes. L'amélioration a été jugée significativement meilleure dans le groupe CsA (76% vs 63% des réponses étaient cotées excellentes ou bonnes dans le groupe CsA comparé au groupe MP). Les chiens traités par la CsA ont présenté significativement plus de troubles gastro-intestinaux (notamment des vomissements) mais les chiens du groupe MP se sont avérés plus sensibles aux infections. Aucune modification remarquable des paramètres hématologiques ou biochimiques n'a été remarquée dans les deux groupes de chiens.

Resumen El objetivo de este estudio multicéntrico, en paralelo, ciego y al azar controlado fue evaluar la eficacia y seguridad de la ciclosporina (grupo CsA, 117 perros) en comparación con la metilprednisolona (grupo MP, 59 perros) en el tratamiento de dermatitis atópica durante 4 meses. La dosis media de inducción para ambas drogas (5 mg kg−1 CsA, 0.75 mg kg−1 MP) fue disminuida con el tiempo de acuerdo con la respuesta clínica. Al final del estudio, el porcentaje de reducción medio estimado desde la línea basal (intervalo de confianza) de la puntuación de las lesiones fue del 52% (44-59) y 45% (35-56), y la reducción de la puntuación del prurito fue del 36% (27-43) y 33% (23-43) en los perros de los grupos de CsA y MP, respectivamente. La diferencia entre los porcentajes de ambos grupos no fue significativa. En la estimación global, la eficacia fue mejor en el grupo de perros tratados con CsA (76% de respuestas excelentes o buenas en el grupo CsA comparado con el 63% en el grupo MP). Los perros tratados con CsA mostraron una mayor frecuencia de alteraciones gastro-intestinales, principalmente vómitos, mientras que los perros tratados con MP presentaron mayor susceptibilidad a infecciones. No se observaron diferencias remarcables en los niveles basales de los parámetros hematológicos y bioquímicos en ambos grupos.