Yellow Fever Virus Maintained by Sabethes Mosquitoes during the Dry Season in Cerrado, a Semiarid Region of Brazil, in 2021.

In recent decades, waves of yellow fever virus (YFV) from the Amazon Rainforest have spread and caused outbreaks in other regions of Brazil, including the Cerrado, a savannah-like biome through which YFV usually moves before arriving at the Atlantic Forest. To identify the vectors involved in the maintenance of the virus in semiarid environments, an entomological survey was conducted after confirmation of yellow fever (YF) epizootics at the peak of the dry season in the Cerrado areas of the state of Minas Gerais. In total, 917 mosquitoes from 13 taxa were collected and tested for the presence of YFV. Interestingly, mosquitoes of the Sabethes genus represented 95% of the diurnal captured specimens, displaying a peak of biting activity never previously recorded, between 4:30 and 5:30 p.m. Molecular analysis identified three YFV-positive pools, two from Sabethes chloropterus-from which near-complete genomes were generated-and one from Sa. albiprivus, whose low viral load prevented sequencing. Sa. chloropterus was considered the primary vector due to the high number of copies of YFV RNA and the high relative abundance detected. Its bionomic characteristics allow its survival in dry places and dry time periods. For the first time in Brazil, Sa. albiprivus was found to be naturally infected with YFV and may have played a role as a secondary vector. Despite its high relative abundance, fewer copies of viral RNA were found, as well as a lower Minimum Infection Rate (MIR). Genomic and phylogeographic analysis showed that the virus clustered in the sub-lineage YFVPA-MG, which circulated in Pará in 2017 and then spread into other regions of the country. The results reported here contribute to the understanding of the epidemiology and mechanisms of YFV dispersion and maintenance, especially in adverse weather conditions. The intense viral circulation, even outside the seasonal period, increases the importance of surveillance and YFV vaccination to protect human populations in affected areas.

Phytochemical prospection, hemagglutinating and insecticidal activity of saline extracts from the seeds of Tamboril (Enterolobium contortisiliquum) Vell. Morong (Fabaceae) on Aedes aegypti (Diptera: Culicidae) / Prospecção fitoquímica, atividade hemaglutinante e inseticida dos extratos salinos das sementes de Tamboril (Enterolobium contortisiliquum) Vell. Morong (Fabaceae) sobre o Aedes aegypti (Diptera: Culicidae)

Abstract This study evaluated the insecticidal activity of crude extracts from Enterolobium contortisiliquum (Vell.) seeds on eggs and larvae of A. aegypti, and also verified the phytochemical profile and the presence of lectins in the extract. The 0.15 M NaCl saline solution was used as the extracting substance. For tests with eggs and larvae, the crude extract was used in its raw form (RCE) and boiled at 100º C for 5 min (BCE). Concentrations of 4.68; 9.37; 18.75; 28.13; 37.13 and 46.89 mg/mL, with distilled water as a negative control. Assays were performed in triplicate. The results were subjected to analysis of variance, Tukey's test and Log-Probit analysis to determine LC50 and LC90. BCE showed better results on eggs than RCE, managing to prevent the hatching of larvae in 81.66% ± 10.40 of treated eggs, at a concentration of 46.89 mg/mL. The LC50 and LC90 were set at 35.95 and 52.67 mg/mL, respectively. In tests with larvae, concentrations of 46.89 and 37.13 mg/mL, for RCE and BCE, caused 100% mortality in 24 hours of exposure. Larval mortality at the other concentrations increased with exposure time extending to 48 h. RCE, at 48 h exposure is the most promising extract on larvae (E = 72.77%, LC90 = 10.86 mg/mL). In RCE, the presence of lectins and secondary metabolites: flavonoids, xanthones and phenols, were detected. The results demonstrate the potential of E. contortisiliquum seed extracts with ovicidal and larvicidal action on A. aegypti.

Resumo Esse estudo avaliou a atividade inseticida dos extratos brutos das sementes de Enterolobium contortisiliquum (Vell.) sobre ovos e larvas do A. aegypti, verificou também o perfil fitoquímico e a presença de lectinas no extrato. A solução salina de NaCl 0,15 M foi utilizada como substância extratora. Para os ensaios com ovos e larvas, o extrato bruto foi utilizado na forma crua (RCE) e fervida a 100º C por 5 min (BCE). Foram testadas as concentrações de 4,68; 9,37; 18,75; 28,13; 37,13 e 46,89 mg/mL, tendo a água destilada como controle negativo. Os ensaios foram realizados em triplicada. Os resultados foram submetidos à análise de variância, Teste de Tukey e análise Log-Probit para determinar CL50 e 90. O BCE apresentou melhores resultados sobre os ovos do que o RCE, conseguindo impedir a eclosão das larvas de 81,66% ± 10,40 dos ovos tratados, na concentração de 46,89 mg/mL. As respectivas CL50 e 90, foram definidas em 35,95 e 52,67 mg/mL, respectivamente. Nos testes com larvas, as concentrações de 46,89 e 37,13 mg/mL, para RCE e BCE, causaram 100% de mortalidade em 24 horas de exposição. A mortalidade larval nas demais concentrações aumentou com o tempo de exposição estendendo-se para 48 h. RCE, com 48 h de exposição é o extrato mais promissor sobre as larvas (E = 72,77%, CL90 = 10,86 mg/mL). Em RCE, a presença de lectinas e os metabólitos secundários: flavonoides, xantonas e fenóis, foram detectadas. Os resultados demonstram o potencial dos extratos das sementes de E. contortisiliquum com ação ovicida e larvicida sobre o A. aegypti.

Categorization by Score of Mosquito Species (Diptera: Culicidae) Related to Yellow Fever Epizootics in Argentina.

The mosquito-borne yellow fever virus (YFV) is the cause of a zoonotic disease, with both sylvatic and urban cycles. Some mosquito species have been associated directly with transmission of the virus in South America, although the importance given to each species varies depending on the bibliography, geographical location, or year of publication. For Argentina, few publications have assessed the involvement of mosquito species, especially those included in the sylvatic cycle. Therefore, the goal of our paper was to gather all the information available in South America and categorize all mosquito species potentially involved in the YFV transmission cycle in Argentina according to incriminating vector criteria. Based on three main characteristics ('Hosts', 'Mosquito vector', and 'YF outbreak'), we generated scales of importance by species, one exclusively for Argentina and another for all of South America, Sabethes albiprivus Theobald, 1903 was the most important species in Argentina; whereas the most important species in South America were Haemagogus janthinomys Dyar 1921, Hg. leucocelaenus Dyar and Shannon, 1924, and Sa. chloropterus Von Humboldt, 1819. Our review highlights the lack of research that evaluates the importance of these species for YFV transmission in Argentina, while serving as a starting point to establish priorities for research on the bionomics and vector status of these species.

Bacterial diversity in Haemagogus leucocelaenus (Diptera: Culicidae) from Vale do Ribeira, São Paulo, Brazil.

INTRODUCTION: Mosquitoes (Diptera: Culicidae) are vectors that transmit numerous pathogens to humans and other vertebrates. Haemagogus leucocelaenus is a mosquito associated with transmission of yellow fever virus. The insect gut harbors a variety of microorganisms that can live and multiply within it, thus contributing to digestion, nutrition, and development of its host. The composition of bacterial communities in mosquitoes can be influenced by both biotic and abiotic factors. The goal of this study was to investigate the bacterial diversity of Hg. leucocelaenus and verify the differences between the bacterial communities in Hg. leucocelaenus from three different locations in the Atlantic tropical rain forest and southeastern state of São Paulo State, Brazil. RESULTS: The phylum Proteobacteria was found in mosquitoes collected from the three selected study sites. More than 50% of the contigs belong to Wolbachia, followed by 5% Swaminathania, and 3% Acinetobacter. The genus Serratia was found in samples from two locations. CONCLUSIONS: Wolbachia was reported for the first time in this species and may indicates that the vector competence of the populations of the species can vary along its geographical distribution area. The presence of Serratia might facilitate viral invasion caused by the disruption of the midgut barrier via action of the SmEnhancin protein, which digests the mucins present in the intestinal epithelium.

Winter survival of Aedes aegypti (Diptera: Culicidae) eggs at its southern limit distribution.

Aedes aegypti (L.) (Diptera: Culicidae) survives in the egg stage under unfavorable environmental conditions. In this study, we assessed the survival of Ae. aegypti eggs under natural winter conditions as an overwintering mechanism. To address this, field collected eggs (from Buenos Aires city) were exposed for three months during the winter season in three sites of Santa Rosa and General Acha (La Pampa province, Argentina). Eggs survival and hatching response were analyzed considering two factors (site of exposure and time of oviposition). Of the 1397 analyzed eggs, 936 (67%) were viable at the end of the study. Egg survival showed to be high in all sites (between 53% and 84%), except in one site of Santa Rosa. Also, eggs survival was higher in eggs laid in May (94%) (late-fall) as compared to those laid in March (61%) and April (56%) (early- and mid-fall respectively). Eggs hatching response was only affected by the time of oviposition, being lower for eggs laid in May (33%) as compared to March (38%) and April (50%). These results provide information regarding the winter resistance of Ae. aegypti at the limit of its distribution in temperate regions. Given the high percentages of survival found in this study, we consider that the low winter temperatures would not be a limitation for its establishment in southern areas.

Determinants of Aedes mosquito density as an indicator of arbovirus transmission risk in three sites affected by co-circulation of globally spreading arboviruses in Colombia, Ecuador and Argentina.

BACKGROUND: The global impact of Zika virus in Latin America has drawn renewed attention to circulating mosquito-borne viruses in this region, such as dengue and chikungunya. Our objective was to assess socio-ecological factors associated with Aedes mosquito vector density as a measure of arbovirus transmission risk in three cities of potentially recent Zika virus introduction: Ibagué, Colombia; Manta, Ecuador; and Posadas, Argentina, in order to inform disease mitigation strategies. METHODS: We sampled Aedes mosquito populations in a total of 1086 households, using indoor and peridomestic mosquito collection methods, including light traps, resting traps, traps equipped with chemical attractant and aspirators. For each sampled household, we collected socio-economic data using structured questionnaires and data on microenvironmental conditions using iButton data loggers. RESULTS: A total of 3230 female Aedes mosquitoes were collected, of which 99.8% were Aedes aegypti and 0.2% were Aedes albopictus. Mean female Aedes mosquito density per household was 1.71 (standard deviation: 2.84). We used mixed-effects generalized linear Poisson regression analyses to identify predictors of Aedes density, using month, neighborhood and country as random-effects variables. Across study sites, the number of household occupants [incidence rate ratio (IRR): 1.08, 95% confidence interval (CI): 1.01-1.14], presence of entry points for mosquitoes into the household (IRR: 1.51, 95% CI: 1.30-1.76) and presence of decorative vegetation (IRR: 1.52, 95% CI: 1.22-1.88) were associated with higher Aedes density; while being in the highest wealth tertile of household wealth (IRR: 0.78, 95% CI: 0.66-0.92), knowledge of how arboviruses are transmitted (IRR: 0.94, 95% CI: 0.89-1.00) and regular emptying of water containers by occupants (IRR: 0.79, 95% CI: 0.67-0.92) were associated with lower Aedes density. CONCLUSIONS: Our study addresses the complexities of arbovirus vectors of global significance at the interface between human and mosquito populations. Our results point to several predictors of Aedes mosquito vector density in countries with co-circulation of multiple Aedes-borne viruses, and point to modifiable risk factors that may be useful for disease prevention and control.

Sexual Competitiveness and Induced Egg Sterility by Aedes aegypti and Aedes albopictus Gamma-Irradiated Males: A Laboratory and Field Study in Mexico.

The sterile insect technique may prove useful for the suppression of mosquito vectors of medical importance in regions where arboviruses pose a serious public health threat. In the present study, we examined the effects of sterilizing irradiation doses across different ratios of fertile:irradiated males on the mating competitiveness of Ae. aegypti and Ae. albopictus under laboratory and field-cage conditions. For both species, the percentage of females inseminated and the number of eggs laid over two gonotrophic cycles varied significantly in mating treatments involving 1:1, 1:5, and 1:10 fertile:irradiated males compared to controls of entirely fertile or entirely irradiated males but was not generally affected by the irradiation dose. Egg hatching was negatively affected in females exposed to increasing proportions of irradiated males in both laboratory and field cages. Male competitiveness (Fried's index) values varied from 0.19 to 0.58 in the laboratory and were between 0.09 and 1.0 in field cages, depending on th species. Competitiveness values were negatively affected by th eirradiation dose in both species under field-cage conditions, whereas in the laboratory, Ae. albopictus was sensitive to the dose but Ae. aegypti was not. In general, male competitiveness was similar across all mating regimes. Most importantly, induced egg sterility was positively correlated with the proportion of irradiated males present in the mating treatments, reaching a maximum of 88% under field-cage conditions for both Ae. aegypti and Ae. albopictus males treated with 50 and 40 Gy irradiation, respectively. These results indicate that sterile males produced at our facility are suitable and competitive enough for field pilot SIT projects and provide guidance to decide the optimal sterile:fertile ratios.

Action of Metarhizium brunneum (Hypocreales: Clavicipitaceae) Against Organophosphate- and Pyrethroid-Resistant Aedes aegypti (Diptera: Culicidae) and the Synergistic Effects of Phenylthiourea.

Dengue, yellow fever, Zika, and chikungunya arboviruses are endemic in tropical countries and are transmitted by Aedes aegypti. Resistant populations of this mosquito against chemical insecticides are spreading worldwide. This study aimed to evaluate the biological effects of exposure of pesticide-sensitive Ae. aegypti larvae (Rockefeller) to conidia of the entomopathogen, Metarhizium brunneum, laboratory strains ARSEF 4556 and V275, and any synergistic activity of phenylthiourea (PTU). In addition, to investigate the nature of any cross-resistance mechanisms, these M. brunneum strains were tested against the Rockefeller larvae and two temephos- and deltamethrin-resistant wild mosquito populations from Rio de Janeiro. Treatment of Rockefeller larvae with 106 conidia/ml of ARSEF 4556 and V275 fungal strains resulted in significant decreased survival rates to 40 and 53.33%, respectively (P < 0.0001), compared with untreated controls. In contrast, exposure to 104 or 105 conidia/ml showed no such significant survival differences. However, the addition of PTU to the conidia in the bioassays significantly increased mortalities in all groups and induced a molt block. Experiments also showed no differences in Ae. aegypti mortalities between the fungal treated, wild pesticide-resistant populations and the Rockefeller sensitive strain. The results show the efficacy of M. brunneum in controlling Ae. aegypti larvae and the synergistic role of PTU in this process. Importantly, there was no indication of any cross-resistance mechanisms between Ae. aegypti sensitive or resistant to pesticides following treatment with the fungi. These results further support using M. brunneum as an alternative biological control agent against mosquito populations resistant to chemical insecticides.

Mosquito abundance in a Dirofilaria immitis hotspot in the eastern state of Rio de Janeiro, Brazil.

Coastal lowlands in the state of Rio de Janeiro, Brazil, support high numbers of heartworm-infected dogs. Microfilariae of heartworm need to be ingested by a potencial mosquito vector in order to develop into infective larvae and infect a new host. Ochlerotatus taeniorhynchus and Ochlerotatus scapularis are the primary vector species in the coastal lowlands of metropolitan Rio de Janeiro; thus, the aim of this study was to investigate whether these two species were abundant enough at the heartworm hotspot in the eastern area of the state to be important to the local parasite's life cycle. The study was conducted at the Massambaba sandbank (22°55'45″S;42°18'51″W), where canine heartworm prevalence was 53.1%. Mosquitoes were captured monthly using two traps with CO2, as well as eventual human landing. A total of 6118 mosquitoes of 16 species were collected. Ochlerotatus taeniorhynchus (37.5%), Culex quinquefasciatus (19.3%), Oc. scapularis (18.2%), and Coquillettidia sp. (10.5%) were the most abundant species. The monthly variation in frequency was marked for Oc. scapularis and Cx. quinquefasciatus and steadier for Oc. taeniorhynchus and Coquillettidia sp. The abundance of the two Ochlerotatus species reaffirms that they are the primary natural vectors for D. immitis in the coastal Rio de Janeiro state lowlands.

Leptolegnia chapmanii (Straminipila: Peronosporomycetes) as a future biorational tool for the control of Aedes aegypti (L.).

The aim of the present review is to summarize the current knowledge about Leptolegnia chapmanii as a pathogen of mosquito larvae. To this end, we present data on its identification, distribution, host range and effects on non-target organisms, effects of environmental factors, in vitro growth, release and persistence in anthropic environments, and effect combined with other insecticides. The data presented allow confirming its potential as a biocontrol agent.

Mayaro Virus in Child with Acute Febrile Illness, Haiti, 2015.

Mayaro virus has been associated with small outbreaks in northern South America. We isolated this virus from a child with acute febrile illness in rural Haiti, confirming its role as a cause of mosquitoborne illness in the Caribbean region. The clinical presentation can mimic that of chikungunya, dengue, and Zika virus infections.

Species-specific escape of Plasmodium sporozoites from oocysts of avian, rodent, and human malarial parasites.

BACKGROUND: Malaria is transmitted when an infected mosquito delivers Plasmodium sporozoites into a vertebrate host. There are many species of Plasmodium and, in general, the infection is host-specific. For example, Plasmodium gallinaceum is an avian parasite, while Plasmodium berghei infects mice. These two parasites have been extensively used as experimental models of malaria transmission. Plasmodium falciparum and Plasmodium vivax are the most important agents of human malaria, a life-threatening disease of global importance. To complete their life cycle, Plasmodium parasites must traverse the mosquito midgut and form an oocyst that will divide continuously. Mature oocysts release thousands of sporozoites into the mosquito haemolymph that must reach the salivary gland to infect a new vertebrate host. The current understanding of the biology of oocyst formation and sporozoite release is mostly based on experimental infections with P. berghei, and the conclusions are generalized to other Plasmodium species that infect humans without further morphological analyses. RESULTS: Here, it is described the microanatomy of sporozoite escape from oocysts of four Plasmodium species: the two laboratory models, P. gallinaceum and P. berghei, and the two main species that cause malaria in humans, P. vivax and P. falciparum. It was found that sporozoites have species-specific mechanisms of escape from the oocyst. The two model species of Plasmodium had a common mechanism, in which the oocyst wall breaks down before sporozoites emerge. In contrast, P. vivax and P. falciparum sporozoites show a dynamic escape mechanism from the oocyst via polarized propulsion. CONCLUSIONS: This study demonstrated that Plasmodium species do not share a common mechanism of sporozoite escape, as previously thought, but show complex and species-specific mechanisms. In addition, the knowledge of this phenomenon in human Plasmodium can facilitate transmission-blocking studies and not those ones only based on the murine and avian models.

Risk factors for West Nile virus infection and disease in populations and individuals.

West Nile virus (WNV) is a mosquito-borne enveloped positive-strand RNA virus that emerged in North America in 1999 in New York City. Over the past 15 years, WNV has become established throughout the USA and has spread into Canada, Mexico and the Caribbean. CDC reports indicate >41,000 clinical cases, including more than 1700 fatalities. An estimated 3 million people in the USA may have been infected to date. Infection with WNV is dependent on many factors including climate, mosquito habitats and immunologically naïve bird populations. In addition, variations within individuals contribute to the risk of severe disease, in particular, advanced age, hypertension, immunosuppression and critical elements of the immune response. Recent advances in technology now allow detailed analysis of complex immune interactions relevant to disease susceptibility.

Chikungunya virus and its mosquito vectors.

Chikungunya virus (CHIKV), a mosquito-borne alphavirus of increasing public health significance, has caused large epidemics in Africa and the Indian Ocean basin; now it is spreading throughout the Americas. The primary vectors of CHIKV are Aedes (Ae.) aegypti and, after the introduction of a mutation in the E1 envelope protein gene, the highly anthropophilic and geographically widespread Ae. albopictus mosquito. We review here research efforts to characterize the viral genetic basis of mosquito-vector interactions, the use of RNA interference and other strategies for the control of CHIKV in mosquitoes, and the potentiation of CHIKV infection by mosquito saliva. Over the past decade, CHIKV has emerged on a truly global scale. Since 2013, CHIKV transmission has been reported throughout the Caribbean region, in North America, and in Central and South American countries, including Brazil, Columbia, Costa Rica, El Salvador, French Guiana, Guatemala, Guyana, Nicaragua, Panama, Suriname, and Venezuela. Closing the gaps in our knowledge of driving factors behind the rapid geographic expansion of CHIKV should be considered a research priority. The abundance of multiple primate species in many of these countries, together with species of mosquito that have never been exposed to CHIKV, may provide opportunities for this highly adaptable virus to establish sylvatic cycles that to date have not been seen outside of Africa. The short-term and long-term ecological consequences of such transmission cycles, including the impact on wildlife and people living in these areas, are completely unknown.