Recently, the endobacteria Wolbachia has emerged as a biological tool for the control of arboviruses. Thus, we investigated the rate of natural infection by Wolbachia in Culicidae species from ...Maranhão, Brazil. For this, we amplified the Wolbachia surface protein gene (wsp) from mosquitoes collected in six localities of Maranhão, and positive samples were subjected to new analysis using group-specific primers. In total, 448 specimens comprising 6 genera and 18 species of mosquitoes were analyzed. Wolbachia DNA was PCR-detected in 7 species, three of which are new records: Aedes scapularis (Rondani, 1848), Coquillettidia juxtamansonia (Chagas, 1907) and Cq. venezuelensis (Theobald, 1912), in addition to Ae. albopictus (Skuse, 1894) and Culex quinquefasciatus Say, 1823, which are commonly described as permissive to maintain this bacterium in natural environments, and two species of the subgenera Anopheles (Nyssorhynchus) Blanchard, 1902 and Culex (Melanoconion) Theobald, 1903 which could not be identified at species level. The infection rate of all species ranged from 0 to 80%, and the average value was 16.5%. This study increases the knowledge about the prevalence of Wolbachia in the culicid fauna and may help in selecting strains for biological control purposes.
Wolbachia pipientis (Hertig) is an endosymbiotic microorganism widespread among arthropods and other invertebrate hosts, and employed in strategies to reduce the incidence of arthropod-borne ...diseases. Here, we used a PCR-based approach for 16S RNA and wsp genes to investigate the prevalence, geographical distribution, and strains of Wolbachia in sand flies (Diptera: Psychodidae: Phlebotominae), the main vectors of the causative agents of leishmaniasis, from three biomes in Brazil: Amazon, Cerrado, and Caatinga. We found that: 1) Wolbachia DNA is present in most (66.7%) of the sampled sand fly species, including vectors of Leishmania spp. (Ross, Trypanosomatida: Trypanosomatidae), 2) the prevalence of Wolbachia DNA varies among species and populations, 3) some strains of Wolbachia may have wider geographical and host range in South America, and 4) two phylogenetic distinct wsp sequences might represent two novel strains for Wolbachia in South America sand flies. Those findings increase the basic knowledge about Wolbachia in South American sand flies and might foster further researches on its use to reduce the transmission of sand fly-borne parasites.
This study aimed to analyze the spatiotemporal distribution of leishmaniases, and contribute to the knowledge of their epidemiological dynamics from 2007 to 2017 in the municipality of Caxias, ...Maranhão, Northeast Brazil. Data on American tegumentary leishmaniasis (ATL) and human visceral leishmaniasis (HVL) were obtained in the Epidemiological Surveillance Sector of Caxias, while data on canine visceral leishmaniasis (CVL) were obtained in the Zoonoses Surveillance Unit. For data analysis and spatial representation of leishmaniasis cases, the geoprocessing of the data was performed, and the geometric features of the state of Maranhão, Caxias, and the disease registration sites were obtained from the shapefile database of the Brazilian Institute of Geography and Statistics. Geostatistics was used to create maps based on the Kernel density method, starting from the points, producing a raster file for each case with several data frames, allowing the instantaneous comparison of the phenomena. During the study period, ATL, HVL, and CVL were reported in Caxias, accounting for 114,304 and 8,498 cases, respectively. The geoprocessing analysis showed that leishmaniasis is widely distributed in the urban area of Caxias. However, there are risk areas for the transmission of these diseases to humans and dogs, associated with deforestation and urban expansion, and may vary over time. Preventive measures must focus on risk areas, including conservation efforts and urban planning, in order to reduce the transmission of leishmaniases.
Biting midges are widespread in Brazilian natural ecosystems. However, deforestation and other activities that impact the environment are reducing natural habitats where biting midges proliferate. ...The objective of this study was to verify whether there is variation in the composition, richness, abundance, and seasonality of biting midges between wild and rural environments, in a forest area with intense deforestation. Biting midges were captured using 6 traps installed at an average height of 1.5 m in the peridomicile, intradomicile, and deciduous seasonal forests, once a month from May 2012 to April 2013. In total, 2,182 specimens of 13 species of the genus Culicoides were captured. Species richness was similar in the intradomicile (13 species), forest (12), and peridomicile (11), but species diversity was greater in the peridomicile (H' = 0.803) compared with the intradomicile (H' = 0.717) and forest (H' = 0.687). The order of species dominance varied between the forest (Culicoides paucienfuscatus Barbosa > Culicoides leopodoi Ortiz > Culicoides foxi Ortiz > Culicoides ignacioi Forattini) and peridomicile + intradomicile habitats (C. paucienfuscatus > C. foxi > C. filariferus Hoffman > C. ignacioi). The activity of these dipterans was strongly influenced by meteorological variables, as biting midges are predominant in the rainy season (80.7% of specimens), when higher rainfall, relative humidity, and lower temperatures prevail. The abundance of biting midges was higher in the peridomicile + intradomicile (83.7% of specimens) compared with the degraded forest (16.3%), a result that reflects the loss of forest habitat due to intense and progressive deforestation.
The synergistic effect of light-emitting diodes (LEDs) and kairomones on the attraction of sand flies to light traps was evaluated. Octenol and lactic acid were used as chemical attractants. Green ...LEDs and the incandescent lamps were used as light attractants. Five CDC-type light traps with the respective combination of attractants (incandescent lamp, incandescent lamp + chemical attractant, green LED, green LED + chemical attractant, and chemical attractant alone without light) were set between 18:00 and 06:00 following a Latin square design. A total of 6,536 sand flies and 16 species were collected. The most frequent species collected was Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera, Psychodidae) accounting for 43.21% of all individuals. Order of success (mean, SD) of lactic acid attractant fly capture was as follows: LED + lactic acid (36.83 ± 4.74), LED alone (34.87 ± 4.61), incandescent lamp + lactic acid (22.80 ± 3.19), incandescent lamp alone (12.67 ± 2.03), and lactic acid (0.46 ± 0.13). Order of success of octenol attractant fly capture was as follows: LED + octenol (37.23 ± 5.61), LED alone (35.77 ± 5.69), incandescent lamp + octenol (18.63 ± 3.28), incandescent lamp alone (14.67 ± 2.86), and octenol alone (1.80 ± 0.65). With exception of lactic acid + incandescent light, chemical synergists played no part in significantly increasing light trap capture of phlebotomine sand flies. However, the use of LEDs, with or without such attractants, provided significantly higher capture compared to the incandescent lamp with or without such chemicals, showing that LEDs are suitable and efficient light sources for surveillance and monitoring of phlebotomine sand flies in Brazil.
The aims of the study were to determine the blood feeding preferences of sandflies and to identify species of Leishmania that infected phlebotomines in Caxias, Maranhão, Brazil, an area that is ...highly endemic for leishmaniasis. Sandflies were captured in light traps located in the peridomiciliary environments of randomly selected houses in urban and rural settings between 1800 and 0600 hours on new moon days between March 2013 and February 2015. DNA extracts from 982 engorged female sandflies were submitted to fragment length polymorphism analysis to identify infecting species of Leishmania, and blood sources were identified for 778 of these specimens. Infection by Leishmania infantum was detected in Lutzomyia longipalpis, Lu. whitmani and Lu. termitophila; L. infantum/L. braziliensis in Lu. longipalpis, Lu. whitmani and Lu. trinidadensis; L. shawi in Lu. longipalpis; L. mexicana in Lu. longipalpis; L. braziliensis in Lu. longipalpis and Lu. whitmani; L. guyanensis in Lu. longipalpis and Lu. termitophila; L. amazonensis in Lu. longipalpis and L. lainsoni or L. naiffi in Lu. longipalpis, while Lu. longipalpis and Lu. trinidadensis were infected with unidentified Leishmania sp. Blood sources were identified in 573 individual phlebotomines and the preferred hosts were, in decreasing order, chicken, dog, rodent and human with lower preferences for pig, horse, opossum and cattle. Lu. longipalpis and Lu. whitmani performed mixed feeding on man, dog and rodent, while Lu. longipalpis was the most opportunistic species, feeding on the blood of all hosts surveyed, but preferably on dog/chicken, dog/rodent and rodent/chicken. Our findings reveal the concomitant circulation of Leishmania species that cause visceral leishmaniasis and tegumentary leishmaniasis in the study area, and explain the occurrence of autochthonous human cases of both clinical forms of leishmaniasis in Caxias, Maranhão. The results support our hypothesis that, in the municipality of Caxias, transmission of Leishmania occurs in close proximity to humans.
In this study, we analyzed species composition, richness, and monthly and seasonal abundance of sand flies in an urban area, municipality Caxias, state Maranhão, endemic for American visceral and ...tegumentary leishmaniasis. Sand flies were caught using Center for Disease Control (CDC) light traps in peridomicile (animal shelter) and extradomicile (forest) environments twice a month for 1 yr. A total of 16,332 specimens belonging to 20 species were captured. The predominant species identified in this study were Lutzomyia longipalpis (Diptera: Psychodidae) (Lutz and Neiva 1912) (43.08%) and Lutzomyia whitmani (Diptera: Psychodidae) (Antunes and Coutinho 1939) (43.02%), which occurred throughout the year (every month). The highest abundance of phlebotomine was recorded during the dry season (53.75%) and in the urban peridomicile environment (59.1%), wherein 17 species were found. In the study area, L. longipalpis and L. whitmani are reported to be effective visceral or tegumentary leishmaniasis vectors, respectively, due to their abundance throughout the year.
Biting midges of the genus Culicoides are small insects associated with the transmission of several pathogens, which requires the correct identification of the species, for implementation of ...effective strategies against these insects. However, many species are difficult to identify only by morphological characters. Therefore, the use of molecular methods can help in the taxonomy and systematics of this group. Here, the DNA barcode approach was evaluated for nine species of Culicoides from the State of Maranhão, Brazil. We generated 39 sequences from a 476 bp (base pairs) fragment of the cytochrome c oxidase subunit I (COI) mitochondrial gene. To assess the usefulness of COI barcodes for the identification of these species, paired genetic distances from intra and interspecific comparisons and phylogenetic trees were generated in MEGA and RAxML/BEAST softwares, respectively. In addition, species delimitation was performed using the PTP, GMYC, and ABGD algorithms. The intra and interspecific genetic distances showed a clear distinction between them, demonstrating that, for the taxa studied, there can hardly be ambiguous identifications with barcodes. In the same sense, the phylogenetic reconstruction resulted in well-supported clades for all morphospecies analyzed.
Sand fly (Diptera: Psychodidae) taxonomy is complex and time-consuming, which hampers epidemiological efforts directed toward controlling leishmaniasis in endemic regions such as northeastern Brazil. ...Here, we used a fragment of the mitochondrial cytochrome c oxidase I (COI) gene to identify sand fly species in Maranhão State (northeastern Brazil) and to assess cryptic diversity occurring at different spatial scales. For this, we obtained 148 COI sequences of 15 sand fly species (10 genera) from Maranhão (fine spatial scale), and joined them to COI sequences from other Brazilian localities (distant about 2,000 km from Maranhão, broad spatial scale) available in GenBank.We revealed cases of cryptic diversity in sand flies both at fine (Lutzomyia longipalpis (Lutz and Neiva) and Evandromyia termitophila (Martins, Falcão and Silva)) and broad spatial scales (Migonemyia migonei (França), Pressatia choti (Floch and Abonnenc), Psychodopygus davisi (Root), Sciopemyia sordellii (Shannon and Del Ponte), and Bichromomyia flaviscutellata (Mangabeira)). We argue that in the case of Bi. flaviscutellata, the cryptic diversity is associated with a putative new species. Cases in which DNA taxonomy was not as effective as morphological identification possibly involved recent speciation and/or introgressive hybridization, highlighting the need for integrative approaches to identify some sand fly species. Finally, we provide the first barcode sequences for four species (Brumptomyia avellari (Costa Lima), Evandromyia infraspinosa (Mangabeira), Evandromyia evandroi (Costa Lima and Antunes), and Psychodopygus complexus (Mangabeira)), which will be useful for further molecular identification of neotropical species.
The aim of this study was to investigate whether temperature, rainfall, and humidity influence the annual distribution of Culicoides Latreille 1809, species abundance and richness in rural areas on ...São Luís Island in the state of Maranhão, Brazil. Biting midges were collected in housing for domestic animals in the peridomestic areas of 10 homes. A CDC light trap was installed in each peridomestic area from 6:00 p.m. to 6:00 a.m. once a month for 12 mo. In total, 23,633 individuals belonging to 26 Culicoides species were captured. The most abundant species were Culicoides diabolicus Hoffman, 1925 (27.43%), C. ignacioi Forattini, 1957 (25.9%), C. flavivenulus Costa Lima, 1937 (15.53%), C. insignis Lutz, 1913 (10.66%), C. filariferus Hoffman, 1939 (6.21%), C. boliviensis Spinelli & Wirth, 1984 (6.07%), C. foxi Ortíz, 1950 (2.83%), and C. leopoldoi Ortíz, 1951 (2.54%). Species richness and abundance were greater during the rainy season (24 species; 88% of the individuals) than during the dry season (18 species; 12% of individuals). However, persistent rain on the days the midges were collected or in the preceding 24 h adversely affected abundance. Mean monthly maximum temperature and rainfall on the day of the collection adversely affected Culicoides abundance and richness.