ABSTRACT
Abstract The aim of the present study was to detect Cercopithifilaria bainae and other tick-borne pathogens and to perform molecular characterization of the tick Rhipicephalus sanguineus s.l. collected from dogs. Ticks (n = 432, including 8 larvae, 59 nymphs, and 365 adults) were sampled from domiciled dogs (n = 73) living in Campo Grande, Mato Grosso do Sul (Midwest Brazil). All ticks were morphologically identified as R. sanguineus. Genomic DNA was extracted in pools (three to five ticks per animal) and was used for definition of R. sanguineus haplotypes (based on 16S rRNA analysis) and pathogen identification (Cercopithifilaria sp., Ehrlichia canis, Anaplasma platys, Hepatozoon canis, Babesia vogeli and Rickettsia spp.). Rhipicephal us sanguineus specimens were identified as haplotypes A and B. DNA of Cercopithifilaria bainae (43.83%; 32/73), Ehrlichia canis (24.65%; 18/73), Anaplasma platys (19.17%; 14/73), and Hepatozoon canis (5.47%; 4/73) was detected. The identity of pathogens was confirmed by DNA sequence analysis. The present study confirms the presence of haplotypes A and B of R. sanguineus in the state of Mato Grosso do Sul and its importance as a vector of several pathogens of veterinary concern. Finally, this is the first report to identify C. bainae in ticks in the Midwestern region of Brazil.
Resumo O objetivo do presente estudo foi detectar Cercopithifilaria bainae e outros patógenos transmitidos por carrapatos e realizar a caracterização molecular do carrapato Rhipicephalus sanguineus s.l. coletado em cães. Carrapatos (n = 432, incluindo 8 larvas, 59 ninfas e 365 adultos) foram amostrados de cães domiciliados (n = 73) residentes no município de Campo Grande, Mato Grosso do Sul (centro-oeste do Brasil). Todos os carrapatos foram identificados morfologicamente como R. sanguineus. O DNA genômico foi extraído em pools (três a cinco carrapatos por animal), seguido pela definição de haplótipos (com base no gene 16S rRNA) e pela investigação de patógenos (Cercopithifilaria sp., Ehrlichia canis, Anaplasma platys, Hepatozoon canis, Babesia vogeli e Rickettsia spp.). Os espécimes coletados foram identificados como haplótipos A e B de R. sanguineus. Foram detectados DNA de Cercopithifilaria bainae (43,83%; 32/73), Ehrlichia canis (24,65%; 18/73), Anaplasma platys (19,17%; 14/73) e Hepatozoon canis (5,47%; 4/73). A identidade dos patógenos foi confirmada por análise de sequência de DNA. O presente estudo confirma a circulação dos haplótipos A e B de R. sanguineus no estado de Mato Grosso do Sul e sua importância como vetor de vários patógenos de interesse veterinário. Finalmente, este é o primeiro relato de C. bainae em carrapatos na região centro-oeste do Brasil.
Subject(s)
Animals , Arachnid Vectors/parasitology , Rhipicephalus sanguineus/parasitology , Dogs/parasitology , Rickettsia/isolation & purification , Rickettsia/genetics , Babesia/isolation & purification , Babesia/genetics , Brazil , RNA, Ribosomal, 16S/genetics , Eucoccidiida/isolation & purification , Eucoccidiida/genetics , Ehrlichia canis/isolation & purification , Ehrlichia canis/genetics , Anaplasma/isolation & purification , Anaplasma/geneticsABSTRACT
Resumen Introducción. En estudios previos se detectó la presencia de Leishmania infantum en Rhipicephalus sanguineus, lo cual planteaba la posibilidad de que R. sanguineus transmitiera la leishmaniasis a una variedad de huéspedes. Objetivo. Identificar Leishmania (Viannia) spp. en garrapatas recolectadas en animales silvestres de una zona endémica para leishmaniasis. Materiales y métodos. Se hicieron 81 extracciones individuales de ADN en las garrapatas recogidas de tres tapires o dantas (Tapirus terrestres) y tres pecaríes de collar (Pecari tajacu) cazados en Madre de Dios, Perú. Las garrapatas recolectadas se identificaron taxonómicamente y se prepararon para la identificación del cinetoblasto (kDNA) de Leishmania (Viannia) spp. mediante reacción en cadena de la polimerasa (PCR), así como de la especie de Leishmania mediante PCR de fusión de alta resolución (High Resolution Melt, HRM). Resultados. Se detectó el kDNA de Leishmania (V) spp. en tres garrapatas silvestres de R. (Boophilus) microplus, Canestrini, 1888, recolectadas en un pecarí de collar cazado en la selva de Madre de Dios. El análisis mediante HRM-PCR evidenció que una de las muestras positivas de kDNA tenía una curva compatible con L. (V) guyanensis. Conclusión. Los resultados evidenciaron la presencia de ADN de L. (V) guyanensis en R. (Boophilus) microplus, probablemente adquirida después de picar al pecarí. Es importante hacer nuevos estudios para aclarar la participación de R. (Boophilus) microplus en la transmisión de la leishmaniasis.
Abstract Introduction: Previous studies identified the presence of Leishmania infantum in Rhipicephalus sanguineus and indicated the possibility that it could transmit leishmaniasis to a variety of hosts. Objective: To identify parasites of Leishmania (Viannia) spp. in ticks collected from wild animals in an endemic area for leishmaniasis. Materials and methods: We performed 81 individual DNA extractions from ticks collected from three Tapirus terrestris and three Pecari tajacu in Madre de Dios, Perú. Ticks were taxonomically identified and they were subsequently prepared to identify Leishmania (Viannia) spp. kDNA by PCR and the species of Leishmania by HRM-PCR. Results: Leishmania (Viannia) kDNA was detected in three wild ticks of the species R. microplus, collected from a collard peccary (P. tajacu) hunted in the forests of Madre de Dios. The HRM-PCR showed that one of the positive samples had a kDNA curve compatible with L. (V) guyanensis. Conclusion: The results showed the presence of L. (V) guyanensis DNA in R. microplus possibly acquired after biting a collarde peccary. Therefore, it is important to design future studies to clarify R. microplus involvement in the transmission of leishmaniasis.
Subject(s)
Animals , Male , Arachnid Vectors/parasitology , Artiodactyla/parasitology , Tick Infestations/veterinary , Leishmania guyanensis/isolation & purification , Rhipicephalus/parasitology , Perissodactyla/parasitology , Peru/epidemiology , Species Specificity , Tick Infestations/parasitology , Disease Reservoirs , Leishmaniasis, Mucocutaneous/transmission , Leishmaniasis, Mucocutaneous/epidemiology , Polymerase Chain Reaction , Leishmania guyanensis/genetics , DNA, Kinetoplast/analysis , Endemic DiseasesABSTRACT
A survey of malaria in Northwest Mindoro, Occidental, Mindoro, Philippines is reported. Three species of human plasmodia were identified from 600 blood films examined. The overall prevalence of malaria was 7% (2.8% P. falciparum, 4.3% P. vivax, 0.7% P. malariae). The prevalence of malaria was highest (24%) among children 0 to 15 years of age and only 4 cases (12%) were found among persons over the age of 15. Males and females were equally infected. Study sites with the highest slide-positivity rate were located in the foothill regions which corresponded with the observed presence of two malaria vectors, Anopheles minimus flavirostris and An. maculatus. These sites appeared to be adequate for future studies of drug-resistance, although large numbers of suitable candidates would not be expected.