Characterization of the bacterial microbiome of Amblyomma scalpturatum and Amblyomma ovale collected from Tapirus terrestris and Amblyomma sabanerae collected from Chelonoidis denticulata, Madre de Dios- Peru.

BACKGROUND: Ticks are arthropods that can host and transmit pathogens to wild animals, domestic animals, and even humans. The microbiome in ticks is an endosymbiotic, pathogenic and is yet to be fully understood. RESULTS: Adult male Amblyomma scalpturatum (A. scalpturatum) and Amblyomma ovale (A. ovale) ticks were collected from Tapirus terrestris (T. terrestris) captured in the rural area of San Lorenzo Village, and males Amblyomma sabanerae were collected from Chelonoidis denticulate (C. denticulate) of the Gamita Farm in the Amazon region of Madre de Dios, Peru. The Chao1 and Shannon-Weaver analyses indicated a greater bacterial richness and diversity in male A. sabanerae (Amblyomma sabanerae; 613.65-2.03) compared to male A. scalpturatum and A. ovale (A. scalpturatum and A. ovale; 102.17-0.40). Taxonomic analyses identified 478 operational taxonomic units representing 220 bacterial genera in A. sabanerae and 86 operational taxonomic units representing 28 bacterial genera in A. scalpturatum and A. ovale. Of the most prevalent genera was Francisella (73.2%) in A. sabanerae, and Acinetobacter (96.8%) in A. scalpturatum and A. ovale to be considered as the core microbiome of A. sabanerae and A. scalpturatum/A. ovale respectively. CONCLUSIONS: We found a high bacterial diversity in male of A. sabanerae collected from C. denticulata showed prevalence of Francisella and prevalence of Acinetobacter in male A. scalpturatum and A. ovale collected from T. terrestris. The greatest bacterial diversity and richness was found in males A. sabanerae. This is the first bacterial metagenomic study performed in A. scalpturatum/A. ovale and A. sabanerae collected from T. terrestris and C. denticulata in the Peruvian jungle.

Quantitative Detection of Leishmania in Rhipicephalus microplus and Amblyomma sabanerae in the Peruvian Amazon Basin.

Leishmaniasis is a disease of public importance with a complex transmission cycle. A quantitative PCR was developed by using the small subunit of the ribosomal RNA gene (SSU rRNA) as a DNA target, which is conserved in all Leishmania species. A TaqMan ® probe was designed to have a high specificity. In all, 22 out of 23 (95.7%) ticks classified as R. microplus tested positive for Leishmania sp. The quantification was between 34.1 and 2197.1 parasites per tick in a range of 12 to 769 fg/uL. In addition, 9 out of 10 (90%) ticks classified as Amblyomma sabanerae tested positive for Leishmania sp. The quantification was between 448.6 and 5428.6 parasites per tick in a range of 157 to 1900 fg/µL. Leishmania sp. was identified in very high percentages in Rhipicephalus microplus and Amblyomma sabanerae from wild Pecari tajacu and Chelonoidis denticulata, in quantities of 34.1 and 5428.6 parasites per arthropod, and this could suggest that the ticks were parasitized by sucking blood from the animals from which they were collected. This is the first report about Leishmania parasites found in wild Rhipicephalus microplus and Amblyomma sabanerae, adding new information about the distribution and epidemiology of the parasite in sylvatic areas.

Characterization of the bacterial microbiome of Rhipicephalus (Boophilus) microplus collected from Pecari tajacu "Sajino" Madre de Dios, Peru.

Ticks are arthropods that can host and transmit pathogens to wild animals, domestic animals, and even humans. The bacterial microbiome of adult (males and females) and nymph Rhipicephalus microplus ticks collected from a collared peccary, Pecari tajacu, captured in the rural area of Botijón Village in the Amazon region of Madre de Dios, Peru, was evaluated using metagenomics. The Chao1 and Shannon-Weaver analyses indicated greater bacterial richness and diversity in female ticks (GARH; 375-4.15) and nymph ticks (GARN; 332-4.75) compared to that in male ticks (GARM; 215-3.20). Taxonomic analyses identified 185 operational taxonomic units representing 147 bacterial genera. Of the 25 most prevalent genera, Salmonella (17.5%) and Vibrio (15.0%) showed the highest relative abundance followed by several other potentially pathogenic genera, such as Paracoccus (7.8%), Staphylococcus (6.8%), Pseudomonas (6.6%), Corynebacterium (5.0%), Cloacibacterium (3.6%), and Acinetobacter (2.5%). In total, 19.7% of the detected genera are shared by GARH, GARM, and GARN, and they can be considered as the core microbiome of R. microplus. To the best of our knowledge, this study is the first to characterize the microbiome of ticks collected from P. tajacu and to report the presence of Salmonella and Vibrio in R. microplus. The pathogenic potential and the role of these bacteria in the physiology of R. microplus should be further investigated due to the possible implications for public health and animal health in populations neighboring the habitat of P. tajacu.

Molecular identification of Bartonella bacilliformis in ticks collected from two species of wild mammals in Madre de Dios: Peru.

OBJECTIVE: To study the presence of Bartonella bacilliformis in ticks collected from two wild mammals in Madre de Dios, Peru. RESULTS: A total of 110 ticks were collected. Among the 43 Amblyomma spp. extracted from the 3 Tapirus terrestris only 3 were positive for B. bacilliformis. In addition, 12 out of the 67 Rhipicephalus (Boophilus) microplus obtained from the 3 Pecari tajacu were positive for B. bacilliformis. For the first time B. bacilliformis have been detected in arthropods other than Lutzomyia spp. Further studies are required to elucidate the possible role of ticks in the spread of South American Bartonellosis.

Co-infection with Bartonella bacilliformis and Mycobacterium spp. in a coastal region of Peru.

OBJECTIVE: This study investigated an outbreak of Bartonellosis in a coastal region in Peru. RESULTS: A total of 70 (n = 70) samples with clinical criteria for the acute phase of Bartonellosis and a positive peripheral blood smear were included. 22.85% (n = 16) cases of the samples were positive for Bartonella bacilliformis by PCR and automatic sequencing. Of those positive samples, 62.5% (n = 10) cases were positive only for B. bacilliformis and 37.5% (n = 6) cases were positive to both Mycobacterium spp. and B. bacilliformis. The symptom frequencies were similar in patients diagnosed with Carrion's disease and those co-infected with Mycobacterium spp. The most common symptoms were headaches, followed by malaise and arthralgia.

Detection of Leishmania (V) guyanensis in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) collected from Pecari tajacu.

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.

Detección de Leishmania (V) guyanensis en ejemplares de Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) recolectados en pecaríes de collar (Pecari tajacu) / Detection of Leishmania (V) guyanensis in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) collected from Pecari tajacu

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.