Valéria Cristina Soares Pinheiro

Frequency, diversity, and productivity study on the Aedes aegypti most preferred containers in the city of Manaus, Amazonas, Brazil

The most preferred containers by Aedes aegypti were studied April and July (rainy and dry periods) in two Manaus neighbourhoods. In all, 2,700 premises and 13,912 containers were examined, most (87%) recorded outdoors. Out of the 13,100 inspected premises, only 1.6% showed to be positive for Aedes aegypti, summing up to 7,916 collected samples. Most frequently found containers outdoors in either neighbourhood regardless of rain or dry period were Bottles flasks and Storage, and indoors, Fixed, Flowerpots, and buckets. Productivity was estimated according to the number of premises and positive containers investigated, showing the actual container groups productivity. Considering both rainy and dry periods outdoors at Praça 14 the groups of Tyre, Flask, Bottle, Construction Equipment and Fixed, had the highest averages respectively. Construction Equipment and Flask groups were the most productive in Coroado in April. Flask, Construction Equipment and Storage groups stood out in July.

Evaluation of the residual effect of temephos on Aedes aegypti (Diptera, Culicidae) larvae in artificial containers in Manaus, Amazonas State, Brazil

Trial tests and container observations were conducted in households to verify the residual effect of temephos in Manaus, Amazonas State, Brazil. Three plastic buckets, three tin cans, and three tires filled with water from an artesian well and larvicide were used in the experiment, with twenty-five third-instar larvae, which remained exposed for 24h, followed by mortality readings. The same types of containers were selected from common households. Collection and counts followed by chemical treatment were carried out on the larvae that were found. Follow-up was performed weekly to verify recolonization by Aedes aegypti. The experiment showed 100% mortality in the plastic buckets until day 90, and 80% in the tin cans until day 30, decreasing from day 45 onwards. Mortality in the tires decreased to 35% in the first month. Household results showed 100% mortality for all containers after 24h and differentiated values in the subsequent readings. Larvae were observed on day 35 in a tin can and on day 21 in a gallon can. There was a large diversity of results in the tires, with recolonization observed from day 7 onwards.

Detection of dengue virus serotype 3 by reverse transcription-polymerase chain reaction in Aedes aegypti (Diptera, Culicidae) captured in Manaus, Amazonas

The detection of dengue virus serotypes from Aedes aegypti in Manaus, state of Amazonas was carried out using the reverse transcription-polymerase chain reaction technique. Fourteen pools out 82 (17.1%) were positive for DENV3, providing a minimal infection rate of 2.1% of all analyzed infected female specimens of three different areas of the city.

Conhecimentos de estudantes sobre Leishmaniose Visceral em escolas públicas de Caxias, Maranhão, Brasil

(2) Explique, resumidamente, exatamente a que corresponde essa função (tendo em vista que a maioria da população não conhece o funcionamento hierárquico da Polícia). Resposta: As principais funções do Diretor Geral de Saúde são: I. Coordenar as Unidades de Saúde da PMERJ que hoje compreendem 2 Hospitais, 4 Policlínicas, localizadas nos bairros de Cascadura, Olaria, São João de Meriti e na Cidade de Campos dos Goitacazes e um Centro de Fisioterapia e Reabilitação; II. Implementar políticas de saúde para o Sistema de Saúde da Polícia Militar; III. Gerir junto ao Comandante Geral e ao Chefe do Estado Maior da Polícia Militar os recursos do Fuspom (Fundo de Saúde da Polícia Militar); IV. Planejar junto ao Comando da Corporação o apoio de Resgate e Remoção nas ações de campo da tropa da PMERJ; V. Planejar, implementar a assistência médica hospitalar aos Policiais Militares e seus dependentes diretos. Este Diretor Geral de Saúde em atenção às questões formuladas como parte da entrevista da Revista Ciência & Saúde Coletiva da Fundação Oswaldo Cruz presta as seguintes informações:

Leishmania infection and blood food sources of phlebotomines in an area of Brazil endemic for visceral and tegumentary leishmaniasis

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.

Variation in Aedes aegypti (Linnaeus) (Diptera, Culicidae) infestation in artificial containers in Caxias, state of Maranhão, Brazil

INTRODUCTION Dengue is a serious public health problem worldwide, with cases reported annually in tropical and subtropical regions. Aedes aegypti (Linnaeus, 1762), the main vector of dengue, is a domiciliary species with high dispersal and survival capacities and can use various artificial containers as breeding sites. We assessed potential container breeding sites of A. aegypti in the municipality of Caxias, Maranhão, Brazil. METHODS In the initial phase, we analyzed 900 properties in 3 neighborhoods during the dry and rainy seasons (August-October 2005 and February-April 2006, respectively). During the second sampling period, September 2006-August 2007, we used 5 assessment cycles for 300 properties in a single neighborhood. RESULTS During the dry and rainy seasons, water-storage containers comprised 55.7% (n = 1,970) and 48.5% (n = 1,836) of the total containers inspected, and showed the highest productivity of immature A. aegypti; we found 23.7 and 106.1 individuals/container, respectively, in peridomicile sites. In intradomicile sites, water-storage containers were also the most important breeding sites with 86.4% (n = 973) and 85.6% (n = 900) of all containers and a mean of 7.9 and 108.3 individuals/container in the dry and rainy seaso-October 2006 (1,342). The highest number of positives (70) was recorded in May, mostly (94%) in storage containers. CONCLUSIONS Storage containers are the principal and most productive A. aegypti breeding sites and are a major contributing factor to the maintenance of this vector in Caxias.

Molecular characterization of the gene profile of Bacillus thuringiensis Berliner isolated from Brazilian ecosystems and showing pathogenic activity against mosquito larvae of medical importance

The occurrence of Aedes aegypti, Culex quinquefasciatus, and mosquitoes of the genus Anopheles potentiate the spread of several diseases, such as dengue, Zika, chikungunya, urban yellow fever, filariasis, and malaria, a situation currently existing in Brazil and in Latin America. Control of the disease vectors is the most effective tool for containing the transmission of the pathogens causing these diseases, and the bacterium Bacillus thuringiensis var. israelensis has been widely used and has shown efficacy over many years. However, new B. thuringiensis (Bt) strains with different gene combinations should be sought for use as an alternative to Bti and to prevent the resistant insects selected. Aiming to identify diversity in the Bt in different Brazilian ecosystems and to assess the pathogenicity of this bacterium to larvae of Ae. aegypti, C. quinquefasciatus, and Anopheles darlingi, Bt strains were obtained from the Amazon, Caatinga (semi-arid region), and Cerrado (Brazilian savanna) biomes and tested in pathogenicity bioassays in third-instar larvae of Ae. aegypti under controlled conditions in the laboratory. The isolates with larvicidal activity to larvae of Ae. aegypti were used in bioassays with the larvae of C. quinquefasciatus and An. darlingi and characterized according to the presence of 14 cry genes (cry1, cry2, cry4, cry10, cry11, cry24, cry32, cry44Aa, cry1Ab, cry4Aa, cry4Ba, cry10Aa, cry11Aa, and cry11Ba), six cyt genes (cyt1, cyt2, cyt1Aa, cyt1Ab, cyt2Aa and cyt2Ba), and the chi gene. Four hundred strains of Bt were isolated: 244 from insects, 85 from Amazon soil, and 71 from the Caatinga biome. These strains, in addition to the 153 strains isolated from Cerrado soil and obtained from the Entomopathogenic Bacillus Bank of Maranhão, were tested in bioassays with Ae. aegypti larvae. A total of 37 (6.7%) strains showed larvicidal activity, with positive amplification of the cry, cyt, and chi genes. The most frequently amplified genes were cry4Aa and cry4Ba, both occurring in 59.4% in these strains, followed by cyt1Aa and cyt2Aa, with 56.7% and 48% occurrence, respectively. Twelve (2.2%) strains that presented 100% mortality within 24h were used in bioassays to estimate the median lethal concentration (LC50) for Ae. aegypti larvae. Two strains (BtMA-690 and BtMA-1114) showed toxicity equal to that of the Bti standard strain, and the same LC50 value (0.003mg/L) was recorded for the three bacteria after 48h of exposure. Detection of the presence of the Bt strains that showed pathogenicity for mosquito larvae in the three biomes studied was possible. Therefore, these strains are promising for the control of insect vectors, particularly the BtMA-1114 strain, which presents a gene profile different from that of Bti but with the same toxic effect.

Occurrence of Aedes aegypti (Diptera, Culicidae) in a Dengue Transmission Area at Coastal Maranhão State, Brazil

This study aimed to estimate the occurrence of Aedes aegypti adults at distinct climatic seasons at neighbor- hoods from the municipality of Sao Luis, Maranhao State, Brazil, as well as to verify the presence of Dengue virus (DENV) in the specimens collected. A total of 320 properties were visited in eight neighborhoods that were previously randomly chosen. Samplings were made at three periods: dry season/2008, rainy season/2009 and dry season/2009. A to- tal of 563 Ae. aegypti mosquitoes were collected, with 141 of them collected during dry season/2008, 272 during rainy season/2009 and the remnant, during dry season/2009. Specimen were divided into lots and subjected to RT-semi-nested- PCR analysis and virus isolation was carried out using cell culture (C6/36 clone) of Ae. albopictus. The number of adults significantly varied at the neighborhoods of Coreia de Baixo, Lira, Joao Paulo, and Conjunto Cohatrac I. Molecular anal- yses of specimens showed no positivity for DENV. At the municipality of Sao Luis, seasonal climate variation might in- fluence the density of Ae. aegypti adults, however, other factors such as population density, sanitation conditions, and the presence of mosquito breeding sites ought to be assessed as important parameters influencing vector dispersion.

Circulation of Chikungunya virus in Aedes aegypti in Maranhão, Northeast Brazil

The simultaneous circulation of Dengue virus (DENV), Chikungunya virus (CHIKV) and Zika virus (ZIKV) arboviruses have placed Brazil among the main worldwide endemic areas. Brazilian Northeast region concentrates the highest incidence of infections caused by CHIKV and ZIKV. In Maranhão, the second biggest northeastern state, there are cases of human infections caused by these three arboviruses and presence of Aedes aegypti and Aedes albopictus vectors. In this context, this study aimed to investigate the circulation of CHIKV, DENV and ZIKV in Ae. aegypti and Ae. albopictus mosquitoes collected in urban areas of Barra do Corda, Caxias, Codó, São Luís and São Mateus do Maranhão municipalities in the state of Maranhão through Quantitative Reverse Transcription Polymerase Chain Reaction (RT-qPCR) technique. 428 Ae. aegypti and 1 Ae. albopictus were collected, which formed 44 pools. Three of these showed positive results for CHIKV: AR832767 (five Ae. aegypti female collected in Caxias), AR832784 and AR832785 (both composed of 20 Ae. aegypti female collected in São Mateus do Maranhão). This study consolidates information about CHIKV circulation in state of Maranhão, as well as the role of Ae. aegypti in the transmission of CHIKV in urban area.

DIVERSIDADE DE ANOPHELES E INFECÇÃO POR MALÁRIA NA ZONA DOS COCAIS, ESTADO DO MARANHÃO, BRASIL

A survey of the Anopheles species was conducted in the municipality of São Mateus, in the state of Maranhão. Species hematophagy habits, the influence of seasonality as well as the vector’s infection by Plasmodium were studied. The female Anopheles mosquitoes were captured between February and December 2010 and in March 2012, from 18h00 to 21h00, in intra and peridomiciles. PCR was used for the infection studies. A total 615 specimens of Anopheles were captured, of which 223 (36.2%) in the locality of Piquizeiro and 392 (63.8%) on the Retiro Velho Farm belonging to the species An. nuneztovari, An. darlingi, An. triannulatus, An. albitarsise, and An. oswaldoi. In Piquizeiro most specimens were captured in the intradomiciliary area, while on the Retiro Velho Farm they were mostly found in the peridomiciliary area. The results show the influence of seasonality and time of capture in the hematophagy frequency of the captured Anopheles. The negative result for Plasmodium infection demonstrates a reduction in the number of malaria cases in the State during the study period.