The relationship among bats, ectoparasites and associated microorganisms is important to investigate how humans can become exposed to zoonotic agents. Even though the diversity of Bartonella spp. in bats and ectoparasites has been previously reported, the occurrence of gltA genotypes within hosts has not been assessed so far. We aimed to investigate the genetic diversity of Bartonella spp. in non-hematophagous bats and associated ectoparasites by assessing cloned gltA Bartonella genotypes in intra- and inter-hosts levels, as well as by using three additional molecular markers. Overall, 13.5% (18/133) bat blood samples, 17.18% bat flies (11/64) and 23.8% (5/21) Macronyssidae mite pools showed to be positive for Bartonella spp. Seventeen positive samples were submitted to gltA-cloning and three clones were sequenced for each sample. We also obtained 11, seven and three sequences for nuoG, rpoB and ftsZ genes, respectively. None were positive for the other target genes. We found at least two genotypes among the three gltA-cloned sequences from each sample, and 13 between all the 51 sequences. Among the nuoG, rpoB and ftsZ sequences we found eight, five and three genotypes, respectively. In the phylogenetic analysis, the sequences were positioned mainly in groups related to Bartonella identified in rodents, bats and bat flies. Herein, we showed the genetic diversity of Bartonella in bat’s blood and associated ectoparasites samples at both intra- and inter-host levels.

The Anaplasmataceae family (order Rickettsiales) encompasses obligately intracellular bacteria of the genera Anaplasma, Ehrlichia, and Neorickettsia. Together with Coxiella burnetii (Coxiellaceae family, order Legionellales), these bacteria represent important causative agents of diseases in humans and animals. The scarcity of studies that investigated the occurrence of these agents in bats and their associated ectoparasites, emphasizes the need to achieve a better understanding of the role of these animals in the maintenance of such bacteria. Herein, 418 samples (133 blood, 135 spleen, and 150 ectoparasites) are collected from 135 non-hematophagous bats belonging to 12 species in a periurban area of Campo Grande city, Mato Grosso do Sul state, midwestern Brazil. In the results, 1.65% (7/418), 12.04% (50/418), and 13.63% (57/418) of samples are positive in PCR assays for Anaplasma spp. (16S rRNA gene), Ehrlichia spp. (dsb gene), and Neorickettsia spp. (16S rRNA gene), respectively. Anaplasma spp. and Neorickettsia spp. are detected in one (5.26%) Ornithodoros hasei tick larva. Ehrlichia spp. is detected in 14% of bat flies (represented by Megistopoda aranea, Trichobius costalimai, and Strebla hertigi), 6% of tick larvae (O. hasei), 12% of Spinturnicidae mites (represented by Periglischrus sp., P. torrealbai, and P. acutisternus), and 38% of Macronyssidae mites (Steatonyssuss sp.). The obtained sequences are observed to be similar to Anaplasma phagocytophilum (97.42–97.6% identified), Ehrlichia minasensis (96.73–100% identified), Neorickettsia risticii (96.7–100% identified), and Neorickettsia findlayensis (95.07–100% identified) by BLASTn analyses, and closely related to Ehrlichia ruminantium by phylogenetic analyses based on the gltA gene. No bat samples (blood/spleen) are positive in the qPCR assay for C. burnetii based on the IS1111 gene. The present work shows, for the first time, the occurrence of Anaplasmataceae in bats and associated ectoparasites (ticks, mites, and bat flies) from Brazil.

Zika virus (ZIKV) was first discovered in 1947 in Uganda but was not considered a public health threat until 2007 when it found to be the source of epidemic activity in Asia. Epidemic activity spread to Brazil in 2014 and continued to spread throughout the tropical and subtropical regions of the Americas. Despite ZIKV being zoonotic in origin, information about transmission, or even exposure of non-human vertebrates and mosquitoes to ZIKV in the Americas, is lacking. Accordingly, from February 2017 to March 2018, we sought evidence of sylvatic ZIKV transmission by sampling whole blood from approximately 2000 domestic and wild vertebrates of over 100 species in West-Central Brazil within the active human ZIKV transmission area. In addition, we collected over 24,300 mosquitoes of at least 17 genera and 62 species. We screened whole blood samples and mosquito pools for ZIKV RNA using pan-flavivirus primers in a real-time reverse-transcription polymerase chain reaction (RT-PCR) in a SYBR Green platform. Positives were confirmed using ZIKV-specific envelope gene real-time RT-PCR and nucleotide sequencing. Of the 2068 vertebrates tested, none were ZIKV positive. Of the 23,315 non-engorged mosquitoes consolidated into 1503 pools tested, 22 (1.5%) with full data available showed some degree of homology to insect-specific flaviviruses. To identify previous exposure to ZIKV, 1498 plasma samples representing 62 species of domestic and sylvatic vertebrates were tested for ZIKV-neutralizing antibodies by plaque reduction neutralization test (PRNT₉₀). From these, 23 (1.5%) of seven species were seropositive for ZIKV and negative for dengue virus serotype 2, yellow fever virus, and West Nile virus, suggesting potential monotypic reaction for ZIKV. Results presented here suggest no active transmission of ZIKV in non-human vertebrate populations or in alternative vector candidates, but suggest that vertebrates around human populations have indeed been exposed to ZIKV in West-Central Brazil.

Studies involving parasitism between ectoparasitic flies and bats are scarce for urban environments, which makes it difficult to find association patterns in these altered environments. Herein, we describe the bat flies community, and estimate the prevalence, mean intensity of infestation, infracommunities, and specificity of bat fliesassociations in urban remnants of Cerrado. We collected 371 bat flies of 14 species and 944 bats of 17 species. The most abundant species of bats were Artibeus planirostris (n = 312), Artibeus lituratus (n = 197), Carollia perspicillata (n = 124) and Platyrrhinus lineatus (n = 110). Regarding bat flies, Megistopoda aranea (n = 148) and Trichobius joblingi (n = 118) were the most abundant. The prevalence of bat flies ranged from 0.3 to 43.5% and the mean infestation intensity from 1 to 12 (mean general of 2.04). We found 16 bat flies infracommunities on the most representative hosts (n > 30). Concerning specificity, 71% of bat flies were associated with a single host species (monoxenic). Our results indicate that bats were most parasitized by a single bat flies species, and prevalence and specificity between bat flies and bats were lower in these urban environments.