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.

Considered ecologically generalist, Didelphis albiventris is reported as reservoir for different species of parasites, especially Trypanosoma cruzi. However, the knowledge about the influence of T. cruzi on hematological parameters of free-living opossum remains scarce. The present study aimed to evaluate the influence of T. cruzi on hematological parameters of white-ear opossums (D. albiventris) from Campo Grande, Mato Grosso do Sul, Brazil. The blood samples and biometric data were collected from 40 opossums captured by Tomahawk and Sherman traps in six urban forest fragments located in the city. The health of these animals was inferred, mainly, by means of blood parameters (PCV, RBC, WBC, MCV and WBC differential). Molecular detection of T. cruzi infection was performed by nested polymerase chain reaction (nPCR), using 18S and 24Sα rDNA region as target. Paired-t-test and simple linear regression were used for statistical analysis. No significant difference was observed between the averages of hematological variables in relation to gender and body condition. The molecular diagnosis showed that 32.5% (13/40) of the opossums were infected by T. cruzi, which presented lymphocytosis (3.4 ± 1.5) and eosinophilia (0.09 ± 0.13). Path analysis showed that T. cruzi infection resulted in increased numbers of lymphocytes and indirectly decreased the body condition of opossums. Moreover T. cruzi infection resulted in a direct effect on decrease of MCV. Overall, our results suggest that T. cruzi infection may represent a risk to health of opossums since the lymphocytosis may cause a secondary damage on body condition of infected animals.