Molecular detection and identification of tick-borne pathogens in Equus caballus and ticks from western Cuba.

Detección e identificación molecular de patógenos transmitidos por garrapatas en Equus caballus y garrapatas del occidente de Cuba.

HORSES; MOLECULAR diagnosis; RICKETTSIA; DERMACENTOR; TICKS; AMBLYOMMA; VECTOR-borne diseases; PATHOGENIC microorganisms; TICK infestations; FELIS

Babesia caballi, Theileria equi y varias especies de rickettsias son agentes de enfermedades transmitidas por vectores que afectan a los equinos. El objetivo del presente estudio fue detectar infecciones por B. caballi y T. equi en caballos e identificar rickettsias en caballos y garrapatas en la región occidental de Cuba. Se estandarizaron 2 ensayos de nPCR para la detección de B. caballi y T. equi. Se colectaron muestras de sangre de caballos, y garrapatas. Se realizó identificación por frotis sanguíneo y detección e identificación molecular de B. caballi, T. equi y Rickettsia sp. Se observaron formaciones intraeritrocíticas compatibles con B. caballi y T. equi. El nPCR mostró que el 25 % de las muestras fueron positivas para B. caballi, 73 % para T. equi y 20 % mostraron coinfección. Los resultados se confirmaron con la secuenciación parcial de los genes bc48 (B. caballi) y ema-1 (T. equi). La secuenciación del gen 18S de ARNr de T. equi demostró la presencia de al menos 2 genotipos de aislados de T. equi en Cuba. El ensayo de PCR en tiempo real y la secuenciación revelaron la presencia de Rickettsia amblyommatis en A. mixtum y Rickettsia felis en D. nitens. Como conclusiones estos resultados constituyen la primera evidencia molecular de B. caballi y T. equi en equinos y el primer reporte de R. felis en D. nitens en Cuba, lo que amplía el conocimiento sobre la distribución de patógenos y el espectro de vectores potenciales contribuyendo al fortalecimiento de los programas de manejo y control. [ABSTRACT FROM AUTHOR]

Babesia caballi, Theileria equi and several species of rickettsias are agents of vector-borne diseases that affect equines. The objective of this study was to detect infections by B. caballi and T. equi in horses and to identify rickettsias in horses and ticks in the western region of Cuba. Two nPCR assays were developed and standardized for the detection of B. caballi and T. equi. Blood samples from horses and ticks were collected. Identification by blood smear and molecular detection and identification of B. caballi, T. equi and Rickettsia spp. were carried out. Intraerythrocytic formations compatible with B. caballi and T. equi were observed. The nPCR showed that 25 % of the samples were positive for B. caballi, 73 % for T. equi and 20 % showed coinfection. The results were confirmed with the partial sequencing of the genes bc48 (B. caballi) and ema-1 (T. equi). The sequencing of the 18S rRNA gene of T. equi demonstrated the presence of at least two genotypes of T. equi isolates in Cuba. The real time qPCR assay and sequencing revealed the presence of Rickettsia amblyommatis in A. mixtum and Rickettsia felis in D. nitens. Conclusions: These results constitute the first piece of molecular evidence of B. caballi and T. equi in horses and the first report of R. felis in D. nitens in Cuba, which broadens the knowledge about the distribution of pathogens and the spectrum of potential vectors contributing to the strengthening of management and control programs. [ABSTRACT FROM AUTHOR]

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