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Hand-held insect nets are the standard method for capturing vector mosquitoes of sylvatic arboviruses; however, occupational risks and biases due to individual skill and attractiveness are important limitations. The use of chemical attractants and automatic traps could be an alternative to resolve these limitations. This study compares the yields achieved using nets with those employing electrical traps with CO2 and BG-Lure®, near the ground and in the canopy strata (6.0 and 8.0 m high). The study was conducted at the Cantareira State Park, which is in the Brazilian Atlantic Forest biome. In the 18 collections performed, 3570 specimens of 52 taxa were obtained. The most frequent species captured near the ground were Wyeomyia confusa and Limatus durhamii, whereas Sabethes albiprivus, Sabethes purpureus, and Haemagogus leucocelaenus were the most frequent in the canopy. The nets resulted in greater species richness and abundance, followed by the trap employing CO2. The combination of CO2 traps with BG-Lure® did not improve performance. The use of BG-Lure® alone resulted in low abundance and a low number of species. Our results demonstrate that the use of traps with CO2 can be complementary to collections with nets; however, for species of epidemiological interest such as those of the genera Haemagogus and Sabethes, especially in the canopy, the net remains the method of choice.

Metagenomic studies of mosquito viromes demonstrated a more diverse composition than just an exclusive composition of pathogenic arboviruses transmitted to humans. In our study, the virome of 866 female mosquitoes collected throughout 2020 at the São Paulo Zoo, located in the city of São Paulo/SP—Brazil, was obtained. Specifically, in this paper, we describe a new virus found by viral RNA extraction and next-generation MiSeq sequencing of a group of 23 specimens of Anopheles (Nys.) strodei. The complete genome with a length of 9709 nucleotides was characterized by a positive orientation and a single strand, with a single large ORF, which encodes a polyprotein of 2987 amino acids. The phylogenetic analysis showed an association with the viral family Iflaviridae and the Riboviria realm. We carried out comparisons with translated sequences of the capsid regions of other iflavirus, and the identities in relation to our sequence were below the minimum limit of 90%, indicating that possibly it is a new species of iflavirus. Our findings contribute to expanding knowledge of virome composition among mosquito species in Brazil and globally. Moreover, we provide a viral genome reference specific to this geographic region and Culicidae family of mosquitoes. This resource facilitates future in silico recognition and assembly of viral genomes within metagenomic datasets.

Zoological gardens in forest areas host a large diversity of vertebrate species (exotic and indigenous, free-living and captive, migrant and resident), resulting in an artificial proximity of animal species that would never share the same environment in natural conditions. The presence of mosquitoes enables the transmission of vector-borne pathogens, as is the case with avian malaria parasites. The mild infections in some bird species may become a serious threat to others that do not possess a natural resistance. Thus, the identification of the potential vectors of these parasites is important for future control of these agents, aiming at the bird species conservation. In this study, we collected over 2000 mosquitoes in Sao Paulo Zoo and analyzed them through molecular methods. Six different mosquito species yielded positive for the targeted parasite DNA. We demonstrated that these culicids had fed mainly on bird species and we reported three mosquito species that have never been previously incriminated as potential vectors of these parasites, enabling the use of more specific measures for vigilance and mosquito control. Avian malaria parasites are widespread parasites transmitted by Culicidae insects belonging to different genera. Even though several studies have been conducted recently, there is still a lack of information about potential vectors of Plasmodium parasites, especially in Neotropical regions. Former studies with free-living and captive animals in Sao Paulo Zoo showed the presence of several Plasmodium and Haemoproteus species. In 2015, a pilot study was conducted at the zoo to collect mosquitoes in order to find out (i) which species of Culicidae are present in the study area, (ii) what are their blood meal sources, and (iii) to which Plasmodium species might they be potential vectors. Mosquitoes were morphologically and molecularly identified. Blood meal source and haemosporidian DNA were identified using molecular protocols. A total of 25 Culicidae species were identified, and 6 of them were positive for Plasmodium/Haemoproteus DNA. Ten mosquito species had their source of blood meal identified, which were mainly birds, including some species that were positive for haemosporidian parasites in the former study mentioned. This study allowed us to expand the list of potential vectors of avian malaria parasites and to improve our knowledge of the evolutionary and ecological relationships between the highly diverse communities of birds, parasites, and vectors present at Sao Paulo Zoo.

Some ecological parameters and the distribution of vectors in the municipality of Eldorado, Vale do Ribeira Region, São Paulo, were studied. Entomological surveys were carried out from September 2019 to March 2021. It was observed that a few ecological parameters, including richness, abundance, diversity, and equitability, were typical of a modified environment, where artificial ecotopes maintain the presence of sandflies throughout the year. A total of 11,668 sandflies were captured. The presence of five taxa were observed in Eldorado, with low diversity and high dominance of Nyssomyia intermedia next to Ny. neivai, which are sympatric species. The results presented reinforce the importance of these species in anthropized areas in the transmission of cutaneous leishmaniasis (CL) agents and the need for entomological monitoring. Psathyromyia pascalei was encountered for the first time in the municipality, expanding the known area of distribution of this species in a modified environment.

The hand-net is the standard method for capturing mosquitoes with sylvatic diurnal activity in disease outbreaks in Brazil. However, occupational risks and biases related to the collectors' abilities and attractiveness are important limitations. In this study, we compared hand-nets with automatic traps (CDC) associated to CO2 and BG-Lure in the Vassununga State Park, a Brazilian Savanna protection area. The collections carried out over 27 days on the ground and the forest canopy. A total of 1555 mosquitoes were obtained in 20 taxa. The diversity index ranged between 1.12 and 1.79 and the dominance index from 0.22 to 0.40. The dominant species on the ground was 'Aedes scapularis' (46.0%), and in the canopy, Hg. 'janthinomys/capricornii' (31.9%). 'Haemagogus leucocelaenus' was rare ('n' = 2). The hand-net resulted in the greatest diversity and abundance of species in both strata, followed by the traps associated with CO2. A low degree of similarity was observed between the hand-net on the ground compared to the other capture methods. The use of BG-Lure alone resulted in a low number of specimens. In conclusion, the hand-net is still the method of choice for collecting arbovirus vectors in the diurnal period, especially yellow fever vectors.

Information systems are essential instruments in managing resources, in the evaluation of the epidemiological situation, and for decision-making at all hierarchical levels. Technological advances have allowed the development of systems that meet these premises. Therefore, it is recommended to consider the optimization of data entry and its immediate georeferencing in order to obtain information in real time. To meet this objective, we describe the application introduction process for the implementation of the digital collection of primary data and its integration with the database through synchronization with the SisaWeb platform (Information System for surveillance and control of Aedes aegypti), developed to meet the needs of the Arbovirus Surveillance and Control Program in the state of São Paulo, Brazil. For this purpose, the application—SisaMob—was conceived in the Android Studio development environment, Google®, following the same guidelines as the traditional collection method. Tablets equipped with the Android® operating system were used. To evaluate the implementation of the application, a semi-structured test was applied. The results highlighted that 774.9% (27) of the interviewees evaluated its use positively and, replacing the standard bulletin, 61.1% (22) of the users considered it regular to excellent. The automatic collection of geographic coordinates represented the greatest innovation in the use of the portable device, with reductions in errors and in the time taken to complete the report in the field. The integration to SisaWeb allowed obtaining information in real-time, being easily presented in tabular and graphic modes and spatially arranged through maps, making it possible to monitor the work at a distance, and allowing preliminary analyses during the data collection process. For the future, we must improve the mechanisms for assessing the effectiveness of information, increase the potential of the tool to produce more accurate analyses, which can direct actions more efficiently.

The identification of mosquito species is necessary for determining the entomological components of disease transmission. However, identification can be difficult in species that are morphologically similar. The cytochrome c oxidase subunit I (COI) DNA barcode region is considered a valuable and reliable diagnostic tool for mosquito species recognition, including those that belong to species complexes. Mansonia mosquitoes are found in forests near swampy areas. They are nocturnal and are highly attracted to light. Hematophagous adult females exhibit aggressive biting behavior and can become infected with and transmit pathogens during their feeding, including some epizootic viruses and avian malaria. In Brazil, twelve Mansonia species have been reported. In a recent study from the São Paulo Zoo in Brazil, three morphologically distinct species were collected and identified, namely: Mansonia (Mansonia) indubitans, Ma. (Man.) pseudotitillans and Ma. (Man.) titillans. However, confirmation of these species by molecular identification was unsuccessful due to a lack of COI sequences in the GenBank database. Thus, this research aimed to describe the COI DNA barcode sequences of some morphologically characterized Mansonia (Man.) species from Brazil and to determine their utility in delimiting species collected from the Atlantic Forest and Brazilian Savanna. Accordingly, we provide tools for the genetic identification of species that play a significant role in pathogen transmission in wildlife and potentially humans. We show that the delimitation of Mansonia species via five different approaches based on COI DNA sequences (BI, NJ, ASAP, bPTP and GMYC) yield basically the same groups identified by traditional taxonomy, and we provide the identification of specimens that were previously identified only up to the subgenus level. We also provide COI sequences from two Mansonia species that were not previously available in sequence databases, Ma. wilsoni and Ma. pseudotitillans, and thus contribute to the ongoing global effort to standardize DNA barcoding as a molecular means of species identification.

Mosquito-borne diseases have a significant public health threat worldwide, with arboviruses accounting for a high proportion of infectious diseases and mortality annually. Brazil, in particular, has been suffering outbreaks of diseases transmitted by mosquito viruses, notably those of the genus, such as dengue, Zika, and chikungunya. Against this background, the São Paulo Zoo is an intriguing ecological niche to explore the virome of mosquitoes, potentially shedding light on the dynamics of arbovirus transmission within a confined setting. In this study, we conducted a comprehensive metagenomic analysis of mosquitoes collected from diverse habitats within the zoo, focusing on the , , and genera. From 1,039 contigs of viral origin, we identified 229 viral species infecting mosquitoes, with the orders , and being the most prevalent and abundant. The difference in virome composition was primarily driven by mosquito host species rather than specific collection sites or trap height. Despite environmental disparities, the virome remained remarkably uniform across different areas of the zoo, emphasizing the strong association between mosquito species and their viral communities. Furthermore, we identified a core virome shared among mosquito species, highlighting potential cross-species transmission events and underscoring the need for targeted surveillance and control measures. These results contribute to our understanding of the interplay between mosquitoes, the environment, and viruses, providing valuable insights for disease intervention strategies in mosquito-borne diseases.

The effect of employing collars impregnated with deltamethrin 4% (DM4) to control canine visceral leishmaniasis (CVL) was evaluated. as were the individual factors associated with this infection. A cohort study that included household dogs was conducted between 2002 and 2006. The presence of pathognomonic signals, peridomiciliary sleep habits and breed were the main factors associated with the infection. The use of DM4 collars contributed to the reduction of CVL with an effectiveness of 66%, and the dogs’ survival rate was greater than 90% at 50 months. In conclusion, the adoption of DM4 collars reduced the number of euthanized canines and in the incidence of CVL, and this reduction was sustained for one year after discontinuing the use of the collar.