Background: Extra-Amazonian autochthonous Plasmodium vivax infections have been reported in mountainous regions surrounded by the Atlantic Forest in Espírito Santo state, Brazil.

DNA barcoding has been an effective tool for species identification in several animal groups. Here, we used DNA barcoding to discriminate between 47 morphologically distinct species of Brazilian sand flies. DNA barcodes correctly identified approximately 90% of the sampled taxa (42 morphologically distinct species) using clustering based on neighbor-joining distance, of which four species showed comparatively higher maximum values of divergence (range 4.23–19.04%), indicating cryptic diversity. The DNA barcodes also corroborated the resurrection of two species within the shannoni complex and provided an efficient tool to differentiate between morphologically indistinguishable females of closely related species. Taken together, our results validate the effectiveness of DNA barcoding for species identification and the discovery of cryptic diversity in sand flies from Brazil.

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BackgroundThe transmission of malaria in the extra-Amazonian regions of Brazil, although interrupted in the 1960s, has persisted to the present time in some areas of dense Atlantic Forest, with reports of cases characterized by particular transmission cycles and clinical presentations. Bromeliad-malaria, as it is named, is particularly frequent in the state of Espírito Santo, with Plasmodium vivax being the parasite commonly recognized as the aetiologic agent of human infections. With regard to the spatial and temporal distances between cases reported in this region, the transmission cycle does not fit the traditional malaria cycle. The existence of a zoonosis, with infected simians participating in the epidemiology, is therefore hypothesized. In the present study, transmission of bromeliad-malaria in Espírito Santo is investigated, based on the complete mitochondrial genome of DNA extracted from isolates of Plasmodium species, which had infected humans, a simian from the genus Allouata, and Anopheles mosquitoes. Plasmodium vivax/simium was identified in the samples by both nested PCR and real-time PCR. After amplification, the mitochondrial genome was completely sequenced and compared with a haplotype network which included all sequences of P. vivax/simium mitochondrial genomes sampled from humans and simians from all regions in Brazil.ResultsThe haplotype network indicates that humans and simians from the Atlantic Forest become infected by the same haplotype, but some isolates from humans are not identical to the simian isolate. In addition, the plasmodial DNA extracted from mosquitoes revealed sequences different from those obtained from simians, but similar to two isolates from humans.ConclusionsThese findings strengthen support for the hypothesis that in the Atlantic Forest, and especially in the state with the highest frequency of bromeliad-malaria in Brazil, parasites with similar molecular backgrounds are shared by humans and simians. The recognized identity between P. vivax and P. simium at the species level, the sharing of haplotypes, and the participation of the same vector in transmitting the infection to both host species indicate interspecies transference of the parasites. However, the intensity, frequency and direction of this transfer remain to be clarified.Electronic supplementary materialThe online version of this article (10.1186/s12936-017-2080-9) contains supplementary material, which is available to authorized users.

Alguns insetos transmissores de doenças procriam exclusivamente nas proximidades das residências. O Aedes aegypti, responsável por epidemias de dengue em cidades brasileiras, representa sério risco também para a febre amarela. Com o insucesso da campanha de erradicação do inseto, justifica-se a busca de criadouros fora do alcance das medidas de controle atualmente adotadas. Na Cidade de Vitória, ES, investigou-se a ocorrência de criadouros de Aedes aegypti na água coletada em bromélias nativas, sobre as rochas. Paralelamente, avaliou-se a infestação predial nas áreas urbanas contíguas. Em quatro das cinco áreas investigadas foram encontradas larvas de culicídeos nas bromélias, sendo que em duas foi identificado Aedes aegypti. A presença dos criadouros em bromélias não guardou relação com a infestação predial nas áreas próximas. Torna-se necessário definir se os criadouros em bromélias constituem focos primários do Aedes aegypti, ou se representam uma conseqüência da elevada infestação urbana.

BackgroundThe hypotheses put forward to explain the malaria transmission cycle in extra-Amazonian Brazil, an area of very low malaria incidence, are based on either a zoonotic scenario involving simian malaria, or a scenario in which asymptomatic carriers play an important role.ObjectivesTo determine the incidence of asymptomatic infection by detecting Plasmodium spp. DNA and its role in residual malaria transmission in a non-Amazonian region of Brazil.MethodsUpon the report of the first malaria case in 2010 in the Atlantic Forest region of the state of Espírito Santo, inhabitants within a 2 km radius were invited to participate in a follow-up study. After providing signed informed consent forms, inhabitants filled out a questionnaire and gave blood samples for PCR, and thick and thin smears. Follow-up visits were performed every 3 months over a 21 month period, when new samples were collected and information was updated.ResultsNinety-two individuals were initially included for follow-up. At the first collection, all of them were clearly asymptomatic. One individual was positive for Plasmodium vivax, one for Plasmodium malariae and one for both P. vivax and P. malariae, corresponding to a prevalence of 3.4% (2.3% for each species). During follow-up, four new PCR-positive cases (two for each species) were recorded, corresponding to an incidence of 2.5 infections per 100 person-years or 1.25 infections per 100 person-years for each species. A mathematical transmission model was applied, using a low frequency of human carriers and the vector density in the region, and calculated based on previous studies in the same locality whose results were subjected to a linear regression. This analysis suggests that the transmission chain is unlikely to be based solely on human carriers, regardless of whether they are symptomatic or not.ConclusionThe low incidence of cases and the low frequency of asymptomatic malaria carriers investigated make it unlikely that the transmission chain in the region is based solely on human hosts, as cases are isolated one from another by hundreds of kilometers and frequently by long periods of time, reinforcing instead the hypothesis of zoonotic transmission.Electronic supplementary materialThe online version of this article (10.1186/s12936-018-2263-z) contains supplementary material, which is available to authorized users.

In Brazil, Dengue (DENV) and Zika (ZIKV) viruses are reported as being transmitted exclusively by Aedes aegypti in urban settings. This study established the vectors and viruses involved in an arbovirus outbreak that occurred in 2019 in a rural area of Espírito Santo state, Brazil. Mosquitoes collected were morphologically identified, sorted in samples, and submitted to molecular analysis for arboviruses detection. Phylogenetic reconstruction was performed for the viral sequence obtained. All 393 mosquitoes were identified as Aedes albopictus. DENV-1 genotype V was present in one sample and another sample was positive for ZIKV. The DENV-1 clustered with viruses that have circulated in previous years in large urban centers of different regions in Brazil. This is the first report of A. albopictus infected by DENV and ZIKV during an outbreak in a rural area in Brazil, indicating its involvement in arboviral transmission. The DENV-1 strain found in the A. albopictus was not new in Brazil, being involved previously in epidemics related to A. aegypti, suggesting the potential to A. albopictus in transmitting viruses already circulating in the Brazilian population. This finding also indicates the possibility of these viruses to disperse across urban and rural settings, imposing additional challenges for the control of the diseases.

This study compares the distribution of anopheline mosquitoes in a malaria-endemic municipality (MAL) and a malaria-free municipality (FREE) in an area of the Atlantic Forest in Brazil. Simultaneous quarterly nightly captures were made in three locations in each municipality. One Shannon light trap (Shannon light traps were home made according to specifications published in Am. J. Trop. Med. 1939; 19: 131-140) (SLT) and five CDC light traps (a kind of automatic trap fed by batteries of 12 V and 7 amp/h, with dry ice as a source of CO2; John W. Hock Company, Gainesville, FL) (CLT) (two in the canopy and three at ground level) were operated from 6:00 p.m. to 6:00 a.m. More specimens were captured in MAL (362 in SLTs and 126 in CLTs) than in FREE (66 in SLTs and 59 in CLTs). For the SLTs, Simpson's dominance index was similar in MAL and FREE (D = 0.15 versus D = 0.203, P > 0.7), but Shannon's diversity index was higher for MAL = 1.969 versus H = 1.641, P < 0.01). For the CLTs, Simpson's dominance index was higher in MAL (D = 0.416 versus 0.2688, P < 0.001), and the Shannon diversity index was higher in FREE (H = 1.5222 versus H = 1.115, P < 0.01). In SLTs, Anopheles (Kerteszia) cruzii s.l. frequencies were higher in MAL (chi2 = 23.39; P = 0.000001). In CLTs, An. cruzii s.l. was present in all strata in MAL but only in the canopy inside the forest in FREE (17 specimens). An. cruzii s.l. represented a higher proportion of anophelines in MAL (chi2 = 31; P < 0.000001). The factors that differed in these two areas were anopheline species density and An. cruzii s.l. abundance and distribution.