Long-Term Trends in Mosquito Vector Populations and Their Impact on Yellow Fever Outbreaks in Atlantic Forest Fragments of Rio De Janeiro, Brazil (2016-2021).

Among all living beings, mosquitoes account for the highest number of human fatalities. Our study aimed to determine mosquito egg abundance fluctuation from 2015 to 2020, in order to observe which years had the highest mosquito vector densities and whether they coincided with yellow fever virus outbreaks in both human and nonhuman primates. The study area included Atlantic Forest fragments in the state of Rio de Janeiro. Studies from the Diptera Laboratory at FIOCRUZ were selected and compared along a timeline period of the field collections. The highest peak in egg abundance from the analyzed studies was observed from 2016 to 2017 and from 2015 to 2016. The lowest egg abundance was during the collection periods from 2018 to 2019 and 2019 to 2020. The species with the highest abundance throughout all the periods of the studies analyzed was Haemagogus leucocelaenus, representing 87% of all epidemiological species identified. The species with the lowest abundance was Hg. Janthinomys, representing only 1%. Monitoring the population of mosquitoes is imperative for disease surveillance, as the rise in specimens of various vector species directly impacts the occurrence of yellow fever cases in both nonhuman primates and human populations.

The role of forest fragmentation in yellow fever virus dispersal.

The intense process of deforestation in tropical forests poses serious challenges for the survival of biodiversity, as well as for the human species itself. This scenario is supported by the increase in the incidence of epidemics of zoonotic origin observed over the last few decades. In the specific case of sylvatic yellow fever (YF), it has already been shown that an increase in the transmission risk of the causative agent (yellow fever virus - YFV) is associated with areas with a high degree of forest fragmentation, which can facilitate the spread of the virus. In this study we tested the hypothesis that areas with more fragmented landscapes and a higher edge density (ED) but a high degree of connectivity between forest patches favor YFV spread. To this end, we used YF epizootics in non-human primates (NHPs) in the state of São Paulo to build direct networks, and used a multi-selection approach to analyze which landscape features could facilitate YFV spread. Our results showed that municipalities with the potential to spread the virus exhibited a higher amount of forest edge. Additionally, the models with greater empirical support showed a strong association between forest edge density and the risk of occurrence of epizootic diseases, as well as the need for a minimum threshold of native vegetation cover to restrict their transmission. These findings corroborate our hypothesis that more fragmented landscapes with a higher degree of connectivity favor the spread of YFV, while landscapes with fewer connections tend to act as dead zones for the circulation of the virus.

Physical Attributes of Tree Holes in the Atlantic Forest Edges: Evaluating Their Association with the Presence and Abundance of Immature Haemagogus leucocelaenus.

Sylvatic yellow fever (SYF) was recently a health issue in Brazil (2016-2019) because transmission was facilitated by a high density of vectors, amplifying hosts, and low vaccine coverage of the human population, especially in urban forests in the Southeast Region of Brazil. Moreover, urban forest edges are more likely to have contact between human and sylvatic vector mosquito populations. Here, we show the association between abiotic and biotic features of tree holes as Haemagogus leucocelaenus rearing sites in Cantareira State Park in Atlantic Forest edges. The analyzed physical features of the tree holes were diameter at breast height, tree hole opening diameter, depth, trunk diameter, tree hole volume, collected volume, height (varying from 0.02 to 4.2 m above ground), and the presence of Culicidae species other than Hg. leucocelaenus. We analyzed 105 positive and 68 negative water samples for larval presence and found no differences between them, suggesting the lack of specific physical characteristics in these categories. Hg. leucocelaenus larval abundance was correlated with the collected volume and opening diameter of tree holes. The tree species that most represented negative breeding sites were Euplassa cantareirae, Guarea macrophylla, Psychotria suterella, and Tibouchina pulchra. Four significant clusters as areas with a high risk of SYV were identified by Get-Ordis spatial analysis. Although Hg. leucocelaenus larvae were found in tree holes with high water levels, their occurrence was regulated by that of other mosquito species. Our findings contribute to clarifying immature vector ecology in tree holes related to human exposure to SYF in urban forest edges.

Occurrence of the Sylvatic Yellow Fever Vector Mosquito Haemagogus leucocelaenus (Diptera: Culicidae) in an Atlantic Forest Fragment of the Touristic State of Rio de Janeiro, Brazil.

The yellow fever virus is estimated to cause 30,000 deaths each year worldwide, with the majority of cases and deaths occurring in Africa. The virus is also endemic to Central and South America, including northern and western Brazil. The sylvatic cycle of the virus is related to wild and rural areas, with nonhuman primates as the primary host and wild mosquitoes, specifically from the genera Haemagogus, as vectors. The diversity of the mosquito community plays a significant role in the increase of pathogen transmission to humans. In the present study, we detected fluctuation in populations of vector mosquitoes using ovitraps for Culicidae egg collection. The study area is a forest fragment of the Atlantic Forest, one of the most threatened biomes in Brazil. This biome has been suffering significant deforestation due to anthropic activity. Worryingly, the proximity of human populations to forest environments increases the risk of spreading disease from forest fragments to urban areas. Our findings showed that the highest egg abundance occurred in December 2019, with a significant difference (p = 0.005) between rainy and dry seasons. Most eggs were collected during the rainy period. Subsequent quantification of specimens from epidemiologically relevant species hatched from field-collected eggs resulted in 1,131 (86%) Haemagogus leucocelaenus (Dyar & Shannon, 1924), 111 (8%) Aedes terrens (Walker, 1856), 47 (4%) Aedes albopictus (Skuse, 1894), and 21 (2%) Haemagogus janthinomys (Dyar, 1921). Finally, we assessed the behavior of different vector species performing oviposition on the same breeding site. The highest correlation coefficient was observed between Ae. albopictus and Ae. terrens (rho = 0.52) concerning other Culicidae species. Therefore, we believe that Culicidae population surveillance is crucial for disease monitoring since the increase in specimens of a number of vector species influences the emergence of yellow fever cases in nonhuman primates and human populations.

Report of Mosquito Vectors of Arboviruses from a Federal Conservation Unit in the Atlantic Forest, Rio de Janeiro State, Brazil.

Arbovirus infections, such as dengue, zika, chikungunya, and yellow fever, are a major public health problem worldwide. As the main vectors, mosquitoes have been classified by the Center for Disease Control and Prevention as one of the deadliest animals alive. In this ecological study, we analyzed the population dynamics of important genera and species of mosquito vectors. Mosquito immatures were collected using ovitraps and at natural breeding sites: bamboos and bromeliads. Adult mosquitoes were captured using CDC traps with CO2, Shannon traps, and manual suction tubes. Collections took place during the rainy and dry seasons from 2019 to 2020 in the Serra dos Órgãos National Park, Rio de Janeiro state, Brazil. The highest number of species was recorded in the ovitraps, followed by CDC and bromeliads. The breeding site with the lowest diversity was bamboo, though it showed the highest level of evenness compared to the other breeding sites. The medically important genera reported were Haemagogus spp., Aedes spp., Culex spp., and Wyeomyia spp. Culicid eggs increased in the rainy season, with a peak in November 2019 and January and February 2020, and lower abundance in the dry season, from September to October 2019. Mosquito eggs had a strong positive correlation (ρ = 0.755) with temperature and a moderate positive correlation (ρ = 0.625) with rainfall. This study shows how environmental variables can influence the ecology of disease-vector mosquitoes, which are critical in the maintenance of arbovirus circulation in a threatened biome within the most densely populated region of Brazil.

The influence of landscape structure on the dispersal pattern of yellow fever virus in the state of São Paulo.

Man-made changes to the landscape play a crucial role in altering the epidemiologic patterns of infectious diseases, mainly as a result of pathogen spillover. Sylvatic yellow fever is ideally suited to modeling of this phenomenon as the risk of transmission of the disease as well as its circulation and dispersal are associated with forest fragmentation. In this study we investigated the temporal dispersal pattern of yellow fever virus (YFV) by means of confirmed cases of epizootics in non-human primates in municipalities in the state of São Paulo where there was no recommendation for vaccination in 2017. We analyzed the resistance to dispersal associated with different classes of land use and the geographic distances between the different locations where epizootics were recorded. The model that best explained the temporal dispersal pattern of YFV in the study area indicated that this was influenced by the geographic distance between collection locations and by the permeability of the forest edges (150 m) at the interface with the following core areas: Water, Agricultural, Non-Forest Formation and Forestry. Water, Agricultural, Urban and Forest core areas and the interfaces between the latter two formed important barriers to circulation of the virus. These findings indicate that fragmentation of vegetation tends to decrease the time taken for pathogens to spread, while conservation of forest areas has the opposite effect.

Natural Infection and Vertical Transmission of Zika Virus in Sylvatic Mosquitoes Aedes albopictus and Haemagogus leucocelaenus from Rio de Janeiro, Brazil.

Zika virus (ZIKV) was recently introduced into the Western Hemisphere, where it is suspected to be transmitted mainly by Aedes aegypti in urban environments. ZIKV represents a public health problem as it has been implicated in congenital microcephaly in South America since 2015. Reports of ZIKV transmission in forested areas of Africa raises the possibility of its dispersal to non-human-modified environments in South America, where it is now endemic. The current study aimed to detect arboviruses in mosquitoes collected from areas with low human interference in Rio de Janeiro, Brazil. Using a sensitive pan-flavivirus RT-PCR, designed to detect the NS5 region, pools of Ae. albopictus and Haemagogus leucocelaenus, were positive for both ZIKV and yellow fever (YFV). Virus RNA was detected in pools of adult males and females reared from field-collected eggs. Findings presented here suggest natural vertical transmission and infection of ZIKV in Hg. leucocelaenus and Ae.albopitcus in Brazil.

Influence of landscape composition and configuration on the richness and abundance of potential sylvatic yellow fever vectors in a remnant of Atlantic Forest in the city of São Paulo, Brazil.

The landscape's structure can play a relevant role in epidemic patterns of arboviruses, influencing factors such as abundance, movement, and dispersal ability in arthropod vectors and vertebrate hosts, besides promoting alterations in the rate of potential infectious contacts between these organisms. In the Americas, yellow fever (YF) exhibits only the sylvatic cycle, in which the virus circulates in sylvatic areas among non-human primates, being transmitted by mosquitoes of the Haemagogus and Sabethes genera. In this study, we investigate some aspects of the landscape in relation to diversity and abundance of culicid species associated with YF transmission. Studies were performed in the Cantareira State Park, a remnant of the Atlantic Forest located in Greater Metropolitan São Paulo, Brazil, where the YF virus circulated recently with dozens of deaths in howler monkeys (Alouatta guariba), in addition to reported human cases. Mosquito collections were carried out monthly from February 2015 to April 2017. Mosquitoes were collected from three sites using battery-powered aspirator (12-volt battery), CDC, and Shannon traps for adults, and suction samplers and entomological spoons in breeding sites to collect immature forms. 703 mosquitoes belonging to 12 species of the Aedini and Sabethini tribes were collected. Aedes scapularis and Psorophora ferox exhibited higher abundance, while Haemagogus leucocelaenus, the main vector of YF in São Paulo state, showed lower abundance in all sampled areas. The site with longer edge between forest area and anthropic area presented more richness and abundance of YF vector species, while the site with larger forest cover area and shorter edges between forest and anthropic areas exhibited an inverse pattern. Statistically significant differences were observed between the composition of potential YF vector species among the investigated sites. Although Hg. leucocelaenus occurred in all sampled sites, the different patterns of distribution and abundance of other mosquitoes such as Aedes scapularis and Psorophora ferox suggest that these species may be involved in the transmission of sylvatic YF in the study area.

Predicting Yellow Fever Through Species Distribution Modeling of Virus, Vector, and Monkeys.

Mapping yellow fever (YF) risk is often based on place of infection of human cases, whereas the circulation between nonhuman primates (NHP) and vectors is neglected. In 2008/2009, YF devastated NHP at the southern limit of the disease in the Americas. In view of the recent expansion of YF in Brazil, we modeled the environmental suitability for YF with data from 2008/2009 epizootic, the distribution of NHP (Alouatta spp.), and the mosquito (Haemagogus leucocelaenus) using the maximum entropy algorithm (Maxent) to define risk areas for YF and their main environmental predictors. We evaluated points of occurrence of YF based on dates of confirmed deaths of NHP in three periods, from October 2008 to: December 2008, March 2009, and June 2009. Variables with greatest influence on suitability for YF were seasonality in water vapor pressure (36%), distribution of NHP (32%), maximum wind speed (11%), annual mean rainfall (7%), and maximum temperature in the warmest month (5%). Models of early periods of the epizootic identified suitability for YF in localities that recorded NHP deaths only months later, demonstrating usefulness of the approach for predicting the disease spread. Our data supported influence of rainfall, air humidity, and ambient temperature on the distribution of epizootics. Wind was highlighted as a predicting variable, probably due to its influence on the dispersal of vectors infected with YF in fragmented landscapes. Further studies on the role of wind are necessary to improve our understanding of the occurrence of YF and other arboviruses and their dispersal in the landscape.

Distribution of the Mosquito Communities (Diptera: Culicidae) in Oviposition Traps Introduced into the Atlantic Forest in the State of Rio de Janeiro, Brazil.

The Atlantic Rainforest of South America is one of the major biodiversity hotspots of the world and serves as a place of residence for a wide variety of Culicidae species. Mosquito studies in the natural environment are of considerable importance because of their role in transmitting pathogens to both humans and other vertebrates. Community diversity can have significant effects on the risk of their disease transmission. The objective of this study was to understand the distribution of mosquito communities using oviposition traps in a region of the Atlantic Forest. Sampling was carried out in Bom Retiro Private Natural Reserve (RPPNBR), located in Casimiro de Abreu, Rio de Janeiro, using oviposition traps, which were set in the forest environment, from October 2015 to December 2016. The canonical correspondence analysis was used to assess the influence of the climatic variables (precipitation, maximum dew point, and direction) throughout the seasons on the population density of the mosquito species. The results showed that population density was directly influenced by climatic variables, which acted as a limiting factor for the mosquito species studied. The climatic variables that were significantly correlated with the density of the mosquito species were precipitation, maximum dew point, and direction. Haemagogus janthinomys was positively correlated with the three climatic variables, whereas Haemagogus leucocelaenus was positively correlated with precipitation and maximum dew point, and negatively correlated with direction.

Haemagogus leucocelaenus and Other Mosquitoes Potentially Associated With Sylvatic Yellow Fever In Cantareira State Park In the São Paulo Metropolitan Area, Brazil

The aim of this work was to investigate whether Haemagogus leucocelaenus and other mosquito species associated with sylvatic transmission of yellow fever virus are present in Cantareira State Park (CSP) in the São Paulo Metropolitan Area (SPMA). From October 2015 to March 2016, adult mosquitoes were captured with the Centers for Disease Control and Prevention traps, manual battery-powered aspirators, and Shannon traps; larvae and pupae were collected in natural and artificial breeding sites. A total of 109 adult mosquito specimens and 30 immature forms belonging to 11 taxonomic categories in 4 genera (Aedes, Psorophora, Sabethes, and Haemagogus) were collected, including Hg. leucocelaenus, the main vector of yellow fever. The entomological findings of the present study indicate that the area is of strategic importance for yellow fever surveillance not only because of the significant numbers of humans and nonhuman primates circulating in CSP and its vicinity but also because it represents a potential route for the disease to be introduced to the SPMA...

Haemagogus leucocelaenus and Other Mosquitoes Potentially Associated With Sylvatic Yellow Fever In Cantareira State Park In the São Paulo Metropolitan Area, Brazil.

The aim of this work was to investigate whether Haemagogus leucocelaenus and other mosquito species associated with sylvatic transmission of yellow fever virus are present in Cantareira State Park (CSP) in the São Paulo Metropolitan Area (SPMA). From October 2015 to March 2016, adult mosquitoes were captured with the Centers for Disease Control and Prevention traps, manual battery-powered aspirators, and Shannon traps; larvae and pupae were collected in natural and artificial breeding sites. A total of 109 adult mosquito specimens and 30 immature forms belonging to 11 taxonomic categories in 4 genera (Aedes, Psorophora, Sabethes, and Haemagogus) were collected, including Hg. leucocelaenus, the main vector of yellow fever. The entomological findings of the present study indicate that the area is of strategic importance for yellow fever surveillance not only because of the significant numbers of humans and nonhuman primates circulating in CSP and its vicinity but also because it represents a potential route for the disease to be introduced to the SPMA.

Isolation of yellow fever virus (YFV) from naturally infectied Haemagogus (Conopostegus) leucocelaenus (diptera, cukicudae) in São Paulo State, Brazil, 2009 / Isolamento do vírus da Febre Amarela de mosquitos naturalmente infectados Haemagogus (Conopostegus) leucocelaenus (Diptera, Culicidae), São Paulo, Brasil, 2009

After detecting the death of Howlers monkeys (genus Alouatta) and isolation of yellow fever virus (YFV) in Buri county, São Paulo, Brazil, an entomological research study in the field was started. A YFV strain was isolated from newborn Swiss mice and cultured cells of Aedes albopictus - C6/36, from a pool of six Haemagogus (Conopostegus) leucocelaenus (Hg. leucocelaenus) mosquitoes (Dyar & Shannon) collected at the study site. Virus RNA fragment was amplified by RT-PCR and sequenced. The MCC Tree generated showed that the isolated strain is related to the South American I genotype, in a monophyletic clade containing isolates from recent 2008-2010 epidemics and epizootics in Brazil. Statistical analysis commonly used were calculated to characterize the sample in relation to diversity and dominance and indicated a pattern of dominance of one or a few species. Hg. leucocelaenus was found infected in Rio Grande do Sul State as well. In São Paulo State, this is the first detection of YFV in Hg. leucocelaenus.

Após a detecção de morte de macacos Bugios (gênero Alouatta) e isolamento do vírus da Febre Amarela (YFV) no município de Buri, Estado de São Paulo, Brasil, foi iniciada uma investigação entomológica em campo. Uma cepa de YFV foi isolada em camundongos recém-nascidos e cultura de células de Aedes albopictus - C6/36, a partir de um lote de seis mosquitos Haemagogus (Conopostegus) leucocelaenus (Hg leucocelaenus) Dyar & Shannon coletados no local de estudo. RNA do vírus foi amplificado por RT-PCR e seqüenciado. A topologia gerada indica que a cepa isolada está relacionada ao genótipo South American I, em clado monofilético englobando isolados recentes de epidemias e epizootias entre 2008 e 2009. Análises estatísticas geralmente usadas caracterizaram a amostra em relação à diversidade e dominância, indicando dominância relativa de uma ou poucas espécies. Hg. leucocelaenus foi detectado infectado também no Rio Grande do Sul. No Estado de São Paulo trata-se da primeira detecção do YFV em Hg leucocelaenus.

Encontro de Haemagogus (Conopostegus) leucocelaenus (Diptera: Culicidae), no Município de Porto Alegre, Estado do Rio Grande do Sul / Finding of Haemagogus (Conopostegus) leucocelaenus (Diptera: Culicidae), in the municipality of Porto Alegre, State of Rio Grande do Sul

Em novembro de 2006, foi realizada uma investigação entomológica numa mata nativa do município de Porto Alegre, Estado do Rio Grande do Sul. O resultado foi a captura de várias espécies Culicidae, incluindo o primeiro registro da presença de Haemagogus leucocelaenus na localidade. Esta espécie é considerada vetora do vírus da febre amarela em alguns municípios do Estado, motivando esta comunicação para alertar sobre o potencial da área para circulação do agente etiológico desta doença.

In November 2006, an entomological investigation was carried out in a native forest in the municipality of Porto Alegre, State of Rio Grande do Sul. The result was the capture of several Culicidae species, including the first recording of the presence of Haemagogus leucocelaenus in this locality. This species is considered to be a vector for the yellow fever virus in some municipalities of this State, and this was the motivation for the present communication, in order to warn regarding the potential of this area for circulation of the etiological agent for this disease.