Sylvatic Transmission Research Articles

First Report of Trypanosoma cruzi Infection in Salivary Gland of Bats from the Peruvian Amazon.

.In the Americas, 8 million people are infected with Chagas disease, and an additional 90 million people are at risk for infection. Little is known about the role bats play in the sylvatic transmission cycle of Trypanosoma cruzi, the parasite causing Chagas disease. Here, we captured bats in the villages of Palmiche, Pachacutec, Nuevo San Martin, and Mayuriaga located in the Datem del Marañon Province in Loreto, Peru. Venous blood samples were collected by cardiac puncture or from the upper extremities, and trypanosomatids were identified by microscopy and molecularly. We collected blood samples from 121 bats on filter paper for molecular studies and 111 slides for microscopic examination of thin and thick blood smears from 16 different bat species. The prevalence of trypanosomatids in all bats species was 34.7% (42/121) and the prevalence of T. cruzi was 4.1% (5/121). In hematophagous bat species, the prevalence of trypanosomatids and T. cruzi was 36.9% (27/73) and 2.7% (2/73), respectively. In non-hematophagous bats, the prevalences of trypanosomatids and T. cruzi were 31.2% (15/48) and 6.2% (3/48), respectively. Also, we confirm the presence of T. cruzi in salivary glands of hematophagous bats Diaemus youngi. These results suggest a sylvatic cycle of trypanosomatid transmission in which bats may harbor infectious T. cruzi parasites that could be transmitted to humans via hematophagous bat bites or salivary contamination by non-hematophagous bats of vegetables consumed by humans.

Evaluation of resting traps to examine the behaviour and ecology of mosquito vectors in an area of rapidly changing land use in Sabah, Malaysian Borneo

BackgroundWidespread deforestation occurring in the tropics is hypothesized to impact the transmission of vector-borne diseases (VBD). Predicting how environmental changes will impact VBD transmission is dependent on understanding the ecology and behaviour of potential vector species outside of domestic settings. However there are few reliable sampling tools for measuring the habitat preference and host choice of mosquito vectors; with almost none suitable for sampling recently blood-fed, resting mosquitoes. This study evaluated the use of two mosquito traps: the resting bucket (RB) and sticky resting bucket (SRB) traps relative to CDC backpack aspiration (CDC) for sampling mosquitoes resting in a range of habitats representing a gradient of deforestation. Eight habitats were selected for sampling around two villages in Kudat District, Malaysian Borneo, to reflect the range of habitats available to mosquitoes in and around human dwellings, and nearby forest habitats where reservoir hosts are present: secondary forest (edge, interior and canopy); plantations (palm and rubber); and human settlements (inside, under and around houses).ResultsOver 31 days, 2243 mosquitoes were collected in 5748 discrete collections. Nine mosquito genera were sampled with Aedes and Culex species being present in all habitats and most abundant. RB and CDC backpack aspiration were most efficient for sampling Culex whereas CDC backpack aspiration and SRB were most efficient for Aedes. Most Aedes identified to species level were Ae. albopictus (91%), with their abundance being highest in forest edge habitats. In contrast, Culex were most abundant under houses. Most blood-fed mosquitoes (76%) were found in human settlements; with humans and chickens being the only blood source.ConclusionsRB and SRB traps proved capable of sampling mosquitoes resting in all sampled habitats. However, sampling efficiency was generally low (c.0.1 per trap per day), necessitating traps to be deployed in high numbers for mosquito detection. None of the traps were effective for sampling zoonotic malaria vectors; however, SRB collected relatively higher numbers of the dengue vector Ae. albopictus. The higher abundance of mosquitoes in forest edge habitats indicates the potential value of these traps for investigating sylvatic dengue transmission. This study has demonstrated the merits in application of simple resting traps for characterising mosquito vector resting behaviour outside of the home.

Identification of White-tailed Deer (Odocoileus virginianus) as a Novel Reservoir Species forTrypanosoma cruziin Texas, USA

Chagas disease, a vector-borne parasitic infection caused by Trypanosoma cruzi, represents a significant source of morbidity and mortality in the Americas. Mammalian reservoir species play a large role in propagating the sylvatic transmission cycle of this disease, and this cycle can spill over, resulting in human infections. Our understanding of the wildlife species implicated in propagating this transmission cycle is incomplete. We investigated white-tailed deer ( Odocoileus virginianus) as a potential novel reservoir for this parasite. Only one of the 314 hunter-harvested deer hearts collected across Texas, was PCR-positive (0.3%) for T. cruzi. This finding has potential implications for deer hunters, because it indicates that there might be a risk of blood-borne transmission during the field-dressing process. Hunters should be strongly encouraged to wear gloves and other personal protective equipment when handling carcasses to prevent exposure to infected blood.

Persistence of Yellow fever virus outside the Amazon Basin, causing epidemics in Southeast Brazil, from 2016 to 2018.

BackgroundYellow fever (YF) is endemic in the Brazilian Amazon Basin, and sporadic outbreaks take place outside the endemic area in Brazil. Since 2016, YF epidemics have been occurring in Southeast Brazil, with more than 1,900 human cases and more than 1,600 epizooties of non-human primates (NHPs) reported until April 2018. Previous studies have demonstrated that Yellow fever virus (YFV) causing outbreaks in 2017 formed a monophyletic group.Methodology/Principal findingsAiming to decipher the origin of the YFV responsible for the recent epidemics, we obtained nucleotide sequences of YFV detected in humans (n = 6) and NHPs (n = 10) from Minas Gerais state during 2017–2018. Next, we performed evolutionary analyses and discussed the results in the light of epidemiological records (official numbers of YFV cases at each Brazilian Federative unit, reported by the Brazilian Ministry of Health). Nucleotide sequences of YFV from Southeast Brazil from 2016 to 2018 were highly conserved and formed a monophyletic lineage (BR-YFV_2016/18) within the genotype South America I. Different clusters were observed within lineage BR-YFV_2016/18, one containing the majority of isolates (from humans and NHPs), indicating the sylvatic transmission of YFV. We also detected a cluster characterized by two synapomorphies (amino acid substitutions) that contained YFV only associated with NHP what should be further investigated. The topology of lineage BR-YFV_2016/18 was congruent with epidemiological and temporal patterns of the ongoing epidemic. YFV isolates detected in 2016, in São Paulo state were located in the most basal position of the lineage, followed by the isolates from Minas Gerais and Espírito Santo obtained in 2017 and 2018. The most recent common ancestor of the lineage BR-YFV_2016/18 dated to 2015 (95% credible intervals = 2014–2016), in a period that was coincident with the reemergence of YFV in the Midwest region of Brazil.ConclusionsThe results demonstrated a single introduction of YFV in the Southeast region and the silent viral circulation before the onset of the outbreaks in 2016. Evolutionary analyses combined with epidemiological records supported the idea that BR-YFV_2016/18 was probably introduced from the Midwest into the Southeast region, possibly in São Paulo state. The persistence of YFV in the Southeast region, causing epidemics from 2016 to 2018, suggests that this region presents suitable ecological and climatic conditions for YFV maintenance during the epidemic and interepidemic seasons. This fact poses risks for the establishing of YF enzootic cycles and epidemics, outside the Amazon Basin in Brazil. YF surveillance and studies of viral dynamics deserve particular attention, especially in Midwest, Southeast and neighbor regions which are the main areas historically associated with YF outbreaks outside the Amazon Basin. YFV persistence in Southeast Brazil should be carefully considered in the context of public health, especially for public health decision-makers and researchers.

Epidemiological and genetic studies suggest a common Leishmania infantum transmission cycle in wildlife, dogs and humans associated to vector abundance in Southeast Spain

Leishmania infantum infection was investigated in 202 wild carnivores, rodents and lagomorphs in Southeast Spain using a real-time PCR (rtPCR) in skin and organ samples, mostly spleen. Lesions compatible with leishmaniosis were not observed in any of the animals. Prevalence defined as the percentage of rtPCR-positive animals was 32% overall, and 45% in foxes (n = 69), 30% in rabbits (n = 80) and stone martens (n = 10), 19% in wood mice (n = 16), 0% in black rats (n = 10) and ranged between 0% and 100% in other minoritarian species including badgers, wild cats, wolves, raccoons, genets and hares. Most infected rabbits were rtPCR-positive in skin and not in spleen samples and the opposite was the case for foxes (p < 0.05). L. infantum prevalence was lowest in spring following months of non-exposure to phlebotomine sand fly vectors, and spatially matched recently estimated Phlebotomus perniciosus vector abundance and the prevalence of subclinical infection in dogs and humans. Prevalence increased with altitude and was greater in drier and less windy South and West compared to the coastal Southeast of the study area (p < 0.05). Genetic diversity of L. infantum from foxes, investigated by PCR-restriction fragment length polymorphisms of kinetoplast DNA, revealed B genotype in all animals, which is frequent in people and dogs in the Iberian Peninsula and Morocco. The study provides further evidence that subclinical L. infantum infection is widespread in wildlife with prevalence depending on environmental factors and that parasite tissue tropism may vary according to host species. Moreover, it suggests that sylvatic and domestic transmission cycles are closely interconnected.

Echinococcus multilocularis and Echinococcus shiquicus in a small mammal community on the eastern Tibetan Plateau: host species composition, molecular prevalence, and epidemiological implications

BackgroundThe eastern part of the Tibetan Plateau is now recognized as an endemic region with the highest reported human infection rates in the world of human alveolar echinococcosis (AE) caused by Echinococcus multilocularis. Existing epidemiological studies on AE have mainly focused on the synanthropic environment, while basic parasitological and ecological aspects in wildlife host species remain largely unknown, especially for small mammal hosts. Therefore, we examined small mammal host species composition, occurrence, and the prevalence of both E. multilocularis and E. shiquicus in Shiqu County (Sichuan Province, China), eastern Tibetan Plateau.ResultsIn total, 346 small mammals from five rodent and one pika species were trapped from four randomly set 0.25 ha square plots. Two vole species, Lasiopodomys fuscus (n = 144) and Microtus limnophilus (n = 44), and the plateau pika (Ochotona curzoniae) (n = 135), were the three most-dominant species trapped. Although protoscoleces of E. multilocularis and E. shiquicus were only observed in L. fuscus and O. curzoniae, respectively, cox1 and nad1 gene DNA of E. shiquicus was detected in all the small mammal species except for Neodon irene, whereas E. multilocularis was detected in the three most-dominant species. The overall molecular prevalence of Echinococcus species was 5.8 (95% CI: 3.3–8.2%) ~ 10.7% (95% CI: 7.4–14.0%) (the conservative prevalence to the maximum prevalence with 95% CI in parentheses), whereas for E. multilocularis it was 4.3 (95% CI: 2.2–6.5%) ~ 6.7% (95% CI: 4.0–9.3%), and 1.5 (95% CI: 0.2–2.7%) ~ 4.1% (95% CI: 2.0–6.1%) for E. shiquicus. The prevalence of both E. multilocularis and E. shiquicus, was significantly higher in rodents (mainly voles) than in pikas. Phylogenetic analyses revealed that Echinococcus haplotypes of cox1 from small mammal hosts were actively involved in the sylvatic and anthropogenic transmission cycles of E. multilocularis in the eastern Tibetan Plateau.ConclusionsIn contrast to previous studies, the current results indicated that rodent species, rather than pikas, are probably more important natural intermediate hosts of E. multilocularis and E. shiquicus in the eastern Tibetan Plateau. Thus, understanding interspecific dynamics between rodents and pikas is essential to studies of the echinococcosis transmission mechanism and human echinococcosis prevention in local communities.

Oropouche Fever: A Review.

Oropouche fever is an emerging zoonotic disease caused by Oropouche virus (OROV), an arthropod transmitted Orthobunyavirus circulating in South and Central America. During the last 60 years, more than 30 epidemics and over half a million clinical cases attributed to OROV infection have been reported in Brazil, Peru, Panama, Trinidad and Tobago. OROV fever is considered the second most frequent arboviral febrile disease in Brazil after dengue fever. OROV is transmitted through both urban and sylvatic transmission cycles, with the primary vector in the urban cycle being the anthropophilic biting midge Culicoides paraensis. Currently, there is no evidence of direct human-to-human OROV transmission. OROV fever is usually either undiagnosed due to its mild, self-limited manifestations or misdiagnosed because its clinical characteristics are similar to dengue, chikungunya, Zika and yellow fever, including malaria as well. At present, there is no specific antiviral treatment, and in the absence of a vaccine for effective prophylaxis of human populations in endemic areas, the disease prevention relies solely on vector control strategies and personal protection measures. OROV fever is considered to have the potential to spread across the American continent and under favorable climatic conditions may expand its geographic distribution to other continents. In view of OROV’s emergence, increased interest for formerly neglected tropical diseases and within the One Health concept, the existing knowledge and gaps of knowledge on OROV fever are reviewed.

Trypanosoma cruzi Transmission Among Captive Nonhuman Primates, Wildlife, and Vectors

Natural infection of captive nonhuman primates (NHPs) with Trypanosoma cruzi (agent of Chagas disease) is an increasingly recognized problem in facilities across the southern USA, with negative consequences for NHP health and biomedical research. We explored a central Texas NHP facility as a nidus of transmission by characterizing parasite discrete typing units (DTU) in seropositive rhesus macaques (Macaca mulatta), identifying the wildlife reservoirs, and characterizing vector infection. In seropositive NHPs, we documented low and intermittent concentrations of circulating T. cruzi DNA, with two DTUs in equal proportions, TcI and TcIV. In contrast, consistently high concentrations of T. cruzi DNA were found in wild mesomammals at the facility, yet rodents were PCR-negative. Strong wildlife host associations were found in which raccoons (Procyon lotor) harbored TcIV and opossums (Didelphis virginiana) harbored TcI, while skunks (Mephitis mephitis) were infected with both DTUs. Active and passive vector surveillance yielded three species of triatomines from the facility and in proximity to the NHP enclosures, with 17% T. cruzi infection prevalence. Interventions to protect NHP and human health must focus on interrupting spillover from the robust sylvatic transmission in the surrounding environment.

Diversity and seasonal abundances of mosquitoes at potential arboviral transmission sites in two different climate zones in Switzerland.

Pathogens of medical or veterinary significance that are transmitted by mosquitoes (Diptera: Culicidae) are (re-)emerging in Europe [e.g. West Nile virus (WNV), Dirofilaria nematodes]. Little is known about the spatiotemporal abundances of mosquito species in Switzerland. Therefore, mosquito population dynamics were investigated, focusing on areas of risk for sylvatic or synanthropic transmission, such as natural sites and suburban sites on either side of the Alpine crest. Repeated collections were made using Centers for Disease Control (CDC) traps, juvenile sampling and ovitrapping. A total of 122 831 mosquito specimens of 21 taxa were identified. Levels of mosquito species richness were similar at suburban sites and in natural zones in Switzerland. Mosquito abundances and seasonality were analysed with generalized linear mixed models based on 382 CDC trap samples (29 454 females) and revealed Aedes annulipes/cantans, Aedes geniculatus, Aedes japonicus, Aedes sticticus, Aedes vexans, Coquillettidia richiardii and Culex pipiens/torrentium as the dominant species overall. Abundances of these species were season-dependent in most cases. There was an effect of site with regard to abundance (higher in natural zones), but not with respect to seasonality. Together with data on vector competence and the host preferences of different species, the present data contribute to assessments of risk for pathogen transmission. For example, both natural and suburban environments seem feasible as sites for amplification cycles of WNV and transmission to mammals.

Evidence for multiple sylvatic transmission cycles during the 2016–2017 yellow fever virus outbreak, Brazil

ObjectivesSince December 2016, Brazil has experienced an unusually large outbreak of yellow fever (YF). Whether urban transmission may contribute to the extent of the outbreak is unclear. The objective of this study was to characterize YF virus (YFV) genomes and to identify spatial patterns to determine the distribution and origin of YF cases in Minas Gerais, Espírito Santo and Rio de Janeiro, the most affected Brazilian states during the current YFV outbreak. MethodsWe characterized near-complete YFV genomes from 14 human cases and two nonhuman primates (NHP), sampled from February to April 2017, retrieved epidemiologic data of cases and used a geographic information system to investigate the geospatial spread of YFV. ResultsAll YFV strains were closely related. On the basis of signature mutations, we identified two cocirculating YFV clusters. One was restricted to the hinterland of Espírito Santo state, and another formed a coastal cluster encompassing several hundred kilometers. Both clusters comprised strains from humans living in rural areas and NHP. Another NHP lineage clustered in a basal relationship. No signs of adaptation of YFV strains to human hosts were detected. ConclusionsOur data suggest sylvatic transmission during the current outbreak. Additionally, cocirculation of two distinct YFV clades occurring in humans and NHP suggests the existence of multiple sylvatic transmission cycles. Increased detection of YFV might be facilitated by raised awareness for arbovirus-mediated disease after Zika and chikungunya virus outbreaks. Further surveillance is required, as reemergence of YFV from NHPs might continue and facilitate the appearance of urban transmission cycles.

Limited Evidence for Infection of Urban and Peri-urban Nonhuman Primates with Zika and Chikungunya Viruses in Brazil.

Chikungunya virus (CHIKV) and Zika virus (ZIKV) emerged in the Americas in 2013. Limited antigenic variability of CHIKV and ZIKV may restrict urban transmission cycles due to population protective immunity. In Africa, sylvatic transmission cycles involving nonhuman primates (NHP) are known for CHIKV and ZIKV, causing cyclic reemergence in humans. To evaluate whether sylvatic cycles can be expected in Latin America, we tested 207 NHP collected between 2012 and 2017 in urban and peri-urban settings in Brazil for infection with ZIKV and CHIKV. No animal tested positive for viral RNA in genus-specific and species-specific reverse transcription-PCR (RT-PCR) assays. In contrast, six animals (2.9%) from the families Atelidae, Callitrichidae, and Cebidae showed ZIKV-specific antibodies and 11 (5.3%) showed CHIKV-specific antibodies in plaque reduction neutralization tests (PRNT). Reactivity was monotypic against either ZIKV or CHIKV in all cases, opposing unspecific virucidal activity of sera. PRNT endpoint titers were low at 1:40 in all NHP, and positive specimens did not correspond to the likely dispersal route and time of introduction of both arboviruses. All antibody-positive samples were therefore tested against the NHP-associated yellow fever virus (YFV) and Mayaro virus (MAYV) and against the human-associated dengue virus (DENV) by PRNT. Two ZIKV-positive samples were simultaneously DENV positive and two CHIKV-positive samples were simultaneously MAYV positive, at titers of 1:40 to 1:160. This suggested cross-reactive antibodies against heterologous alphaviruses and flaviviruses in 24% of ZIKV-positive/CHIKV-positive sera. In sum, low seroprevalence, invariably low antibody titers, and the distribution of positive specimens call into question the capability of ZIKV and CHIKV to infect New World NHP and establish sylvatic transmission cycles. IMPORTANCE Since 2013, Zika virus (ZIKV) and chikungunya virus (CHIKV) have infected millions of people in the Americas via urban transmission cycles. Nonhuman primates (NHP) are involved in sylvatic transmission cycles maintaining ZIKV and CHIKV in the Old World. We tested NHP sampled during 2012 to 2017 in urban and peri-urban areas severely affected by ZIKV and CHIKV in Brazil. Seroprevalence and antibody titers were low for both viruses. Additionally, we found evidence for infection by heterologous viruses eliciting cross-reactive antibodies. Our data suggest that urban or peri-urban NHP are not easily infected by ZIKV and CHIKV despite intense local transmission. These data may imply that the ZIKV and CHIKV outbreaks in the Americas cannot be sustained in urban or peri-urban NHP once human population immunity limits urban transmission cycles. Investigation of diverse animals is urgently required to determine the fate of the ZIKV and CHIKV outbreaks in the Americas.

Q fever epidemic in Cayenne, French Guiana, epidemiologically linked to three-toed sloth.

A Q fever epidemic occurred in 2013 in a small military residential area in Cayenne, French Guiana. A retrospective cohort study was conducted to identify Q fever risk factors. Confirmed acute Q fever case was defined as positive serology (IgM ≥ 50 and phase II IgG ≥ 200) and/or positive qPCR on serum or blood. In addition, wild mammals were captured at the study site and tested by serology and real-time PCR performed on blood, vaginal swabs and ticks. The attack rate was 20 percent (11/54). All the cases were symptomatic with fever >38.5 °C and community-acquired pneumonia for four cases. Log binomial multivariate models identified two independent risk factors associated with Q fever: to clean the house (RRa = 7.5 CI95% [1.03-55.3]) and to carry a three-toed sloth in arms (RRa = 2.6 CI95% [1.1-5.8]). Eighteen marsupial individuals were captured, all PCRs were negative but 17% (3/18) had a positive serology. Another study conducted after the epidemic found only one (1/4) three-tooth sloth (Bradypus tridactylus) with feces highly infectious for C. burnetii MST17. The same strain C. burnetii genotype 17 has been laboratory- confirmed in this mammal and in human cases. These results support the implication of three-toed-sloth in this epidemic. Human contamination mainly occurs through inhalation of infectious aerosols as suggested by high relative risk associated with house cleaning activities and pulmonary forms of the disease, and through direct contact with three- toed-sloth. Positive serological results among marsupials confirm wildlife exposure and suggest a more complex sylvatic transmission cycle among wild mammals.

Molecular Characterization of Rabies Virus from Foxes in the Sultanate of Oman

Rabies was firstly recorded in Oman in August 1990, when a school boy had been bitten by a fox. Within a year, then the virus was recorded among red foxes (<i>Vulpes vulpes</i>). Unlike other developing countries, dog is not the usual agent of rabies transmission in Oman but fox considered the main reservoir (sylvatic transmission). The present study was Carried out to determine the molecular epidemiology of fox rabies virus in Oman. The nucleoprotein (N) genes were amplified from brain of three rabid foxes obtained during (2009-2012). The nucleotide sequences of the genes were subjected to molecular and phylogenic analysis. The phylogenic analysis was performed in relation to Omani foxes isolates and other sequences from the Saudi Arabia (isolated in 1987) available in the GenBank. Results indicated that only one genetic group of fox rabies virus appeared to be circulating in Oman since 1990. The study also showed that the recent isolate (2012) of Omani rabies virus from foxes is closely related (97%) to those viruses isolated from foxes in Saudi Arabia. According to the obtained results, It is clear that since its first diagnosis in 1990 the rabies fox virus has not gone through significant virus variant in Oman, It seems that rabies is maintained endemically in Oman through wild foxes. In conclusion the findings reported here highlight the remarkable genetic stability of rabies virus.

The Asia-Pacific origins of the current outbreaks of Zika virus

Zika virus (ZIKV) is a mosquito-borne arbovirus from the Flaviviridae family, first isolated in 1947 from a monkey in Uganda. In the ensuing decades up to the 2000s, there have been sporadic reports of infections and seropositivity in humans in Africa and Asia1,2. The first isolation of ZIKV outside Africa was from Aedes aegypti mosquitoes in Malaysia in 19663. Seropositivity has also been reported in wild monkeys in Malaysia3, although the relevance of this in sylvatic transmission of ZIKV is unknown. These studies suggest that there was endemic and mostly undetected transmission in Asia during this period. Re-emergence from Asia has now brought this relatively neglected virus into the focus of global attention.

Experimental Zika Virus Infection of Neotropical Primates.

The establishment of a sylvatic reservoir of Zika virus (ZIKV) in the Americas is dependent on the susceptibility of primates of sufficient population density, the duration and magnitude of viremia, and their exposure to the human mosquito-borne transmission cycle. To assess the susceptibility of squirrel (Saimiri sp.) and owl monkeys (Aotus sp.) to infection, we inoculated four animals of each species with ZIKV from the current epidemic. Viremia in the absence of detectible disease was observed in both species and seroconversion occurred by day 28. ZIKV was detected in the spleen of three owl monkeys: one at 7 days postinoculation (dpi) and two at 14 dpi. This study confirms the susceptibility to ZIKV infection of two Neotropical primate species that live in close proximity to humans in South America, suggesting that they could support a widespread sylvatic ZIKV cycle there. Collectively, establishment of a ZIKV sylvatic transmission cycle in South America would imperil eradication efforts and could provide a mechanism for continued exposure of humans to ZIKV infection and disease.

Detection of Dengue Virus in Bat Flies (Diptera: Streblidae) of Common Vampire Bats, Desmodus rotundus, in Progreso, Hidalgo, Mexico.

Blood-feeding arthropods play a major role in the transmission of several flaviviruses, which represent an important problem for human health. Currently, dengue is one of the most important arboviral emerging diseases worldwide. Furthermore, some previous studies have reported the presence of viral nucleic acids and antibodies against dengue virus (DENV) in wild animals. Our knowledge of the role played by wildlife reservoirs in the sylvatic transmission and maintenance of DENV remains limited. Our objective was to screen blood-feeding ectoparasites (bat flies) and their common vampire bat (Desmodus rotundus) hosts, for flaviviruses in Hidalgo, Mexico. We detected Flavivirus sequences in 38 pools of ectoparasites (Diptera: Streblidae, Strebla wiedemanni and Trichobius parasiticus) and 8 tissue samples of D. rotundus by RT-PCR and semi-nested PCR using FlaviPF1S, FlaviPR2bis, and FlaviPF3S primers specific for NS5, a gene highly conserved among flaviviruses. Phylogenetic inference analysis performed using the maximum likelihood algorithm implemented in PhyML showed that six sequences clustered with DENV (bootstrap value = 53.5%). Although this study supports other reports of DENV detection in bats and arthropods other than Aedes mosquitoes, the role of these ectoparasitic flies and of hematophagous bats in the epidemiology of DENV still warrants further investigation.

Chikungunya Outbreak in Kedougou, Southeastern Senegal in 2009–2010

BackgroundIn Senegal, Chikungunya virus (CHIKV), which is an emerging mosquito-borne alphavirus, circulates in a sylvatic and urban/domestic cycle and has caused sporadic human cases and epidemics since 1960s. However, the real impact of the CHIKV sylvatic cycle in humans and mechanisms underlying its emergence still remains unknown.MethodologyOne thousand four hundred nine suspect cases of CHIKV infection, recruited from 5 health facilities located in Kedougou region, south-eastern Senegal, between May 2009 to March 2010, together with 866 serum samples collected from schoolchildren from 4 elementary schools in May and November 2009 from Kedougou were screened for anti-CHIKV immunoglobulin (Ig)M antibodies and, when appropriate, for viral nucleic acid by real-time polymerase chain reaction (rPCR) and virus isolation. In addition, mosquitoes collected in the same area from May 2009 to January 2010 were tested for CHIKV by rPCR and by virus isolation, and 116 monkeys sera collected from March 2010 to May 2010 were tested for anti-CHIKV IgM and neutralizing antibodies.ResultsThe main clinical manifestations of the CHIKV suspect cases were headache, myalgia, and arthralgia. Evidence for CHIKV infection was observed in 1.4% (20 of 1409) of patients among suspect cases. No significant difference was observed among age or sex groups. In addition, 25 (2.9%) students had evidence of CHIKV infection in November 2009. Chikungunya virus was detected in 42 pools of mosquitoes, mainly from Aedes furcifer, and 83% of monkeys sampled were seropositive.ConclusionsOur findings further documented that CHIKV is maintained in a sylvatic transmission cycle among monkeys and Aedes mosquitoes in Kedougou, and humans become infected by exposure to the virus in the forest.

High risk for chikungunya virus to initiate an enzootic sylvatic cycle in the tropical Americas.

BackgroundChikungunya virus (CHIKV) has dispersed in the Americas since 2013, and its range of distribution has overlapped large forested areas. Herein, we assess vector competence of two sylvatic Neotropical mosquito species, Haemagogus leucocelaenus and Aedes terrens, to evaluate the risk of CHIKV to initiate a sylvatic cycle in the continent.Methodology/Principal findingsHaemagogus leucocelaenus and Ae. terrens from the state of Rio de Janeiro, Brazil were orally challenged with the two CHIKV lineages circulating in the Americas. Fully engorged females were kept in incubators at 28±1°C and 70±10% humidity and examined at 3 and 7 days after virus exposure. Body (thorax plus abdomen), head and saliva samples were analyzed for respectively determining infection, dissemination and transmission. Both Hg. leucocelaenus and Ae. terrens exhibited high infection and dissemination rates with both CHIKV isolates at 7 dpi, demonstrating that they are susceptible to CHIKV, regardless of the lineage. Remarkably, Hg. leucocelaenus expectorated infectious viral particles as rapidly as 3 days after the infectious blood meal, displaying higher values of transmission rate and efficiency than Ae. terrens. Nevertheless, both species were competent to experimentally transmit both CHIKV genotypes, exhibiting vector competence similar to several American Aedes aegypti.Conclusions/SignificanceThese results point out the high risk for CHIKV to establish a sylvatic transmission cycle in the Americas, which could be a serious health issue as CHIKV would become another zoonotic infection difficult to control in the continent.

Hosts and vectors of Trypanosoma cruzi discrete typing units in the Chagas disease endemic region of the Paraguayan Chaco.

Active Trypanosoma cruzi transmission persists in the Gran Chaco region, which is considered hyperendemic for Chagas disease. Understanding domestic and sylvatic transmission cycles and therefore the relationship between vectors and mammalian hosts is crucial to designing and implementing improved effective control strategies. Here we describe the species of triatomine vectors and the sylvatic mammal reservoirs of T. cruzi, in different localities of the Paraguayan and Bolivian Chaco. We identify the T. cruzi genotypes discrete typing units (DTUs) and provide a map of their geographical distribution. A total of 1044 triatomines and 138 sylvatic mammals were captured. Five per cent of the triatomines were microscopically positive for T. cruzi (55 Triatoma infestans from Paraguay and one sylvatic Triatoma guasayana from Bolivia) and 17 animals (12·3%) comprising eight of 28 (28·5%) Dasypus novemcinctus, four of 27 (14·8%) Euphractus sexcinctus, three of 64 (4·7%) Chaetophractus spp. and two of 14 (14·3%) Didelphis albiventris. The most common DTU infecting domestic triatomine bugs was TcV (64%), followed by TcVI (28%), TcII (6·5%) and TcIII (1·5%). TcIII was overwhelmingly associated with armadillo species. We confirm the primary role of T. infestans in domestic transmission, armadillo species as the principal sylvatic hosts of TcIII, and consider the potential risk of TcIII as an agent of Chagas disease in the Chaco.

Culex (Melanoconion) panocossa from peninsular Florida, USA

Culex (Melanoconion) panocossa is a suspected vector of Venezuelan equine encephalitis virus in Central America. Prior to this report, Cx. panocossa was known from Central America (Belize, Costa Rica, El Salvador, Guatemala, southern Mexico, Panama), northern South America (Colombia, Venezuela) and the Greater Antilles (Cuba and Jamaica). Larvae (n=5) and adults (n=4286) of Cx. panocossa were collected at two locations near Homestead, FL, which indicates substantial established populations of this probable vector species in the continental US. Since larvae of Cx. panocossa are associated with Pistia spp. (water lettuce), the distribution of this mosquito is likely to expand in Florida, where water lettuce is a major invasive plant in freshwater ecosystems. The putative establishment of Cx. panocossa in Florida is of significant concern from a public health perspective, as its proliferation in developed areas could link historically sylvatic transmission foci of Everglades virus with populated centers such as the greater Miami Metropolitan area.