Detección molecular de patógenos emergentes de importancia médica y veterinaria en garrapatas capturadas sobre caballos domésticos / Molecular detection of emerging pathogens of medical and veterinary importance in ticks found in domestic horses

En la actualidad son varias las especies de patógenos emergentes de importancia médica y veterinaria transmitidos por garrapatas. Los estudios sobre estos agentes y sus enfermedades han sido escasos en Cuba. Conocer la presencia de algunos de estos patógenos en garrapatas cubanas que afectan el ganado equino. Se procesaron 95 garrapatas colectadas de caballos domésticos, conservadas en alcohol e identificadas taxonómicamente según claves convencionales. A cada una se le realizó extracción de ADN y posteriormente diferentes reacciones en cadena de la polimerasa utilizando cebadores específicos para los grupos microbianos Borrelia burgdorferi sensu lato, Anaplasma-Ehrlichia, y Babesia-Theileria. Cada uno de los productos de las reacciones en cadena de la polimerasa fue sometido a hibridaciones en línea reversa utilizando sondas para cada grupo en cuestión, así como específicas para las principales especies de estos. Las garrapatas estudiadas pertenecían a las especies Dermacentor (Anocentor) nitens (60 por ciento), Amblyomma cajennense (38 por ciento) y Rhipicephalus (Boophilus) microplus (2 por ciento). Se detectaron 7 garrapatas Dermacentor (Anocentor) nitens infectadas con bacterias del grupo Anaplasma/Ehrlichia, y no se pudo identificar la especie en cuestión con las sondas utilizadas. Una de estas garrapatas estaba además coinfectada con Babesia bovis. Se sugiere la circulación de una nueva especie de Anaplasma o Ehrlichia no reportada antes en Cuba, por lo que se necesita estudiar un número mayor de garrapatas, así como la incorporación de nuevas sondas en la hibridación en línea reversa u otras metodologías que permitan conocer con exactitud las especies que pudiesen afectar hoy día los caballos domésticos.

At present, there are several tick-borne emerging pathogen species of medical and veterinary importance. Few studies on these agents and its diseases have been made in Cuba. To determine the presence of some of these pathogens in Cuban ticks existing in the equine cattle. Ninety five ticks collected from domestic use horses were processed, preserved in alcohol and taxonomically identified according to the set classifications. Their DNA was extracted and subjected to several polymerase chain reactions with specific primers for microbial groups Borrelia burgdorferi sensu lato, Anaplasma-Ehrlichia, y Babesia-Theileria. Each of the products from polymerase chain reactions underwent reverse line blot hybridation using probes for each group as well as specific probes for the main species included in these groups. The studied ticks belonged to Dermacentor (Anocentor) nitens (60 percent), Amblyomma cajennense (38 percent) y Rhipicephalus (Boophilus) microplus (2 percent). Seven Dermacentor (Anocentor) nitens ticks infested with Anaplasma/Ehrlichia bacteria were detected but the species in question could not be detected by the used probes. One of these ticks was also co-infested with Babesia bovis. It is suggested that a new species of Anaplasma o Ehrlichia, not reported in Cuba before now, is circulating, so studying a higher number of ticks is needed and new probes in reverse line blot hybridation or other methodologies must be incorporated to allow exactly determining the species that may affect the Cuban domestic horses at present.

Evaluation of a modified culture medium for Borrelia burgdorferi sensu lato

The aim of the present study was to assess the possible use of a modified medium, prepared in the laboratory using the constituents of Barbour-Stonner-Kelly (BSK) medium and medium 199 as base, for the culture of Borrelia strains, comparing the growth of individual strains in this medium and in the BSK-H medium, and the protein profile and antigenic characteristics of Borrelia proteins expressed in these media. A qualitative evaluation of growth of Borrelia species was made with acceptable results (morphology and motility), but during a quantitative evaluation using the three main genospecies of Borrelia, the better results were obtained with a B. burgdorferi sensu stricto strain. The modified medium did not enable the growth of a B. afzelii strain. The protein profile and antigenic characteristic of the expressed proteins in the modified medium were studied with satisfactory results. These results suggest the modified medium as an alternative for the cultivation of Borrelia strains, with some limitations, in poorly-resourced laboratories.