Flight height preference for oviposition of mosquito (Diptera: Culicidae) vectors of sylvatic yellow fever virus near the hydroelectric reservoir of Simplício, Minas Gerais, Brazil.

In this study, the oviposition behavior of mosquito species exhibiting acrodendrophilic habits was investigated. The study was conducted near the Simplicio Hydroelectic Reservoir (SHR) located on the border of the states of Minas Gerais and Rio de Janeiro, Brazil. Samples were collected using oviposition traps installed in forest vegetation cover between 1.70 and 4.30 m above ground level during the months of April, June, August, October, and December of 2011. Haemagogus janthinomys (Dyar), Haemagogus leucocelaenus (Dyar and Shannon), Aedes albopictus (Skuse), and Aedes terrens (Walker) specimens were present among the collected samples, the first two of which being proven vectors of sylvatic yellow fever (SYF) in Brazil and the latter is a vector of dengue in mainland Asia. As the data set was zero-inflated, a specific Poisson-based model was used for the statistical analysis. When all four species were considered in the model, only heights used for egg laying and months of sampling were explaining the distribution. However, grouping the species under the genera Haemagogus Williston and Aedes Meigen showed a significant preference for higher traps of the former. Considering the local working population of SHR is very large, fluctuating, and potentially exposed to SYF, and that this virus occurs in almost all Brazilian states, monitoring of Culicidae in Brazil is essential for assessing the risk of transmission of this arbovirus.

Cohabitation with aggressive hosts: description of a new microhisterid species in nests of a ponerine ant with ecological notes.

A new clown beetle species, Bacanius neoponerae, is described from Mexican nests of the arboreal ponerine ant Neoponera villosa found in the tank bromeliad Aechmea bracteata. Adult beetles were found in brood chambers or inner refuse piles, but also outside the ant nests, in decaying organic matter between the bromeliad leaves. No direct interactions between ants and microhisterid beetles could be observed. Several lines of evidence suggest a close relationship either with the ants, specific microhabitats within the ant nests or the bromeliads. Sample site elevation, colony size, monthly rainfall and collecting site were the main variables predicting the association. Almost half of the N. villosa colonies were associated with the microhisterids, and larger colonies favored their presence, especially during the driest months of the year. Two specimens were found in a nest of another ant species, Camponotus atriceps, also inhabiting A. bracteata. The new species is the seventh of the genus Bacanius reported from Mexico. This is the second time a species of this genus is associated with ants, and the fourth record of a histerid beetle cohabiting with ponerine ants. The small size of these beetles and their very protective body structure may facilitate their cohabitation with such aggressive hosts.

Yellow fever virus envelope protein expressed in insect cells is capable of syncytium formation in lepidopteran cells and could be used for immunodetection of YFV in human sera.

BACKGROUND: Yellow fever is an haemorrhagic disease caused by a virus that belongs to the genus Flavivirus (Flaviviridae family) and is transmitted by mosquitoes. Among the viral proteins, the envelope protein (E) is the most studied one, due to its high antigenic potencial. Baculovirus are one of the most popular and efficient eukaryotic expression system. In this study a recombinant baculovirus (vSynYFE) containing the envelope gene (env) of the 17D vaccine strain of yellow fever virus was constructed and the recombinant protein antigenicity was tested. RESULTS: Insect cells infected with vSynYFE showed syncytium formation, which is a cytopathic effect characteristic of flavivirus infection and expressed a polypeptide of around 54 kDa, which corresponds to the expected size of the recombinant E protein. Furthermore, the recombinant E protein expression was also confirmed by fluorescence microscopy of vSynYFE-infected insect cells. Total vSynYFE-infected insect extracts used as antigens detected the presence of antibodies for yellow fever virus in human sera derived from yellow fever-infected patients in an immunoassay and did not cross react with sera from dengue virus-infected patients. CONCLUSIONS: The E protein expressed by the recombinant baculovirus in insect cells is antigenically similar to the wild protein and it may be useful for different medical applications, from improved diagnosis of the disease to source of antigens for the development of a subunit vaccine.

Multivariate discrimination between two cryptic Haemagogus species associated with the transmission of yellow fever virus in the Americas.

Mosquitoes of the genus Haemagogus are important vectors of yellow fever virus and other arboviruses and are the principal reservoirs of these viruses in nature. Haemagogus capricornii and Haemagogus janthinomys are closely related species between which females are morphologically cryptic. A morphometric study of these species was performed on male and female specimens from 14 municipalities in Brazil. Morphometric analyses were able to distinguish females. Multivariate morphometrics may be a useful tool for taxonomic studies of cryptic species in this group.

Scanning electron microscopy of the egg of Haemagogus tropicalis.

Haemagogus tropicalis is strictly a forest-dwelling species from the fertile valley area of the Amazônia forest. It is a diurnal mosquito, and the oviposition sites for the species include tree holes. The eggs of Hg. tropicalis used in this study were from females captured on Combú Island, situated across from the city of Belém, Guajará Bay, state of Pará, at 1 degrees 25'S latitude and 48 degrees 25'W longitude. The eggs are elliptical and approximately 575 microm long with a width of approximately 144 microm. The ventral surface of the chorionic reticulum has regular chorionic cells with hexagonal and sometimes pentagonal ornamentation. Each chorionic cell has a thick external chorionic reticulum with regular borders. The interior of the chorionic cells have small, evenly distributed tubercles, and the dorsal external chorionic reticulum appears porous. The micropylar apparatus, located on the anterior area of the egg, was formed by a collar with a well-developed frame. Centrally, the micropylar disc had a diameter of approximately 20 microm and the micropylar orifice is 2.1 microm in diameter. These data may enable construction of taxonomic keys for identifying eggs of Haemagogus species.

Contribution to the knowledge of Saprinus Erichson, 1834 of forensic relevance from Lebanon (Coleoptera, Histeridae).

Many histerid beetles are necrophilous on carrion during both active and advanced stages of decomposition. In this study, 13 species of Saprinus were recorded on carrion from Lebanon, containing eight that are new for the Lebanese fauna. The following Saprinus species are newly recorded from Lebanon: 1) Saprinus (S.) caerulescens caerulescens (Hoffmann, 1803); 2) S. (S.) calatravensis Fuente, 1899; 3) S. (S.) chalcites (Illiger, 1807); 4) S. (S.) godet (Brullé, 1832); 5) S. (S.) maculatus (P. Rossi, 1792); 6) S. (S.) strigil Marseul, 1855; 7) S. (S.) submarginatus J. Sahlberg, 1913; and 8) S. (S.) tenuistrius sparsutus Solsky, 1876. The peak activity was recorded, key for the species is provided, and habitus images and male genitalia are illustrated in order to facilitate their taxonomic identifications. Saprinus species are diverse and common on animal carcass; they were likewise collected from a human cadaver in Lebanon. Preliminary comments on biology and distribution of the studied species are given. Our paper represents the first faunistic study on Histeridae from Lebanon. A rigorous research program regarding the biology of Saprinus in Lebanon and the neighbouring countries would greatly improve the knowledge of the diversity, activity, and possible forensic value of Saprinus.

Influence of spatial heterogeneity on an emerging infectious disease: the case of dengue epidemics.

The importance of spatial heterogeneity and spatial scales (at a village or neighbourhood scale) has been explored with individual-based models. Our reasoning is based on the Chilean Easter Island (EI) case, where a first dengue epidemic occurred in 2002 among the relatively small population localized in one village. Even in this simple situation, the real epidemic is not consistent with homogeneous models. Conversely, including contact heterogeneity on different scales (intra-households, inter-house, inter-areas) allows the recovery of not only the EI epidemiological curve but also the qualitative patterns of Brazilian urban dengue epidemic in more complex situations.

Scanning electron microscopy study of the egg of Haemagogus (Haemagogus) capricornii Lutz, 1904 (Diptera: Culicidae).

Morphological details are provided for the dorsal and ventral surfaces of both extremities and the micropylar area of eggs of Haemagogus (Haemagogus) capricornii Lutz, captured in the Biological Reserve of Tinguá, State of Rio de Janeiro, Brazil. The eggs were observed by scanning electron microscopy with a morphometrical analysis of the main structures. The outer chorionic cells on the ventral surface were extremely regular, such as those observed in Hg. equinus and Hg. janthinomys. The tubercles present differences in form, size, and distribution. Filaments to attach to the substrate were observed in this species.

New species of Omalodes and redefinition of the tribe Omalodini (Coleoptera: Histeridae: Histerinae).

A new species of Omalodini, Omalodes atacamanus sp. nov., is described from Chile. A redefinition of Omalodini is proposed and an identification key to the genera is provided. The new species can be distinguished from other species of Omalodes mainly by all tibiae having a row of setae on outer submarginal region. Omalodini sensu nov. is herein restricted to a lineage of Ebonius Lewis, 1885 and Omalodes Dejean, 1833. The tribe has known distribution across Latin America with some species reaching the southern USA.

Socio-economic and Climate Factors Associated with Dengue Fever Spatial Heterogeneity: A Worked Example in New Caledonia.

BACKGROUND/OBJECTIVES: Understanding the factors underlying the spatio-temporal distribution of infectious diseases provides useful information regarding their prevention and control. Dengue fever spatio-temporal patterns result from complex interactions between the virus, the host, and the vector. These interactions can be influenced by environmental conditions. Our objectives were to analyse dengue fever spatial distribution over New Caledonia during epidemic years, to identify some of the main underlying factors, and to predict the spatial evolution of dengue fever under changing climatic conditions, at the 2100 horizon. METHODS: We used principal component analysis and support vector machines to analyse and model the influence of climate and socio-economic variables on the mean spatial distribution of 24,272 dengue cases reported from 1995 to 2012 in thirty-three communes of New Caledonia. We then modelled and estimated the future evolution of dengue incidence rates using a regional downscaling of future climate projections. RESULTS: The spatial distribution of dengue fever cases is highly heterogeneous. The variables most associated with this observed heterogeneity are the mean temperature, the mean number of people per premise, and the mean percentage of unemployed people, a variable highly correlated with people's way of life. Rainfall does not seem to play an important role in the spatial distribution of dengue cases during epidemics. By the end of the 21st century, if temperature increases by approximately 3 °C, mean incidence rates during epidemics could double. CONCLUSION: In New Caledonia, a subtropical insular environment, both temperature and socio-economic conditions are influencing the spatial spread of dengue fever. Extension of this study to other countries worldwide should improve the knowledge about climate influence on dengue burden and about the complex interplay between different factors. This study presents a methodology that can be used as a step by step guide to model dengue spatial heterogeneity in other countries.

[Scanning electron microscopy of eggs of Haemagogus leucocelaenus (Diptera: Culicidae)]. / Microscopia eletrônica de varredura de ovos de Haemagogus leucocelaenus (Diptera: Culicidae).

OBJECTIVE: To observe morphological details of the eggs of Haemagogus (Conopostegus) leucocelaenus, seen for the first time via scanning electron microscopy (SEM), with morphometric analysis of the main structures. METHODS: Eggs of Hg. leucocelaenus were obtained from females captured in the Biological Reserve of Tinguá, State of Rio de Janeiro. Some of the eggs were kept for hatching and others underwent processing for scanning electron microscopy studies. Three eggs were submitted to morphometric analysis. The material was fixed in 2.5% glutaraldehyde and postfixed in 1% osmium tetroxide, both in 0.1M, pH 7.2 sodium cacodylate buffer, then dehydrated in ethanol and dried using the critical point method. This was then set up on metallic supports, covered with gold and observed using the Jeol 5310 scanning electron microscope. Measurements were made with the aid of the Semafore analysis software coupled to the electron microscope. RESULTS: The eggs presented elliptical outlines of approximately 574 mm in length and 169 mm in width, with an egg index (l/w ratio) of 3.39 mm. The exochorion was extremely regular and had ornamentation that was usually hexagonal but sometimes pentagonal. Tubercles were observed on the chorionic cells, symmetrically arranged in relation to the longitudinal axis. Inside the cells, there were smaller, individualized tubercles, some arranged peripherally and others grouped to a greater or lesser extent in the center. The surface of the chorionic reticulum did not present rugosity. The micropylar apparatus was formed by a prominent continuous collar of 8.32 mm in thickness, with a slightly irregular surface. The micropylar disk was very evident, and was continuous with the collar. The micropyle was seen at the center of this disk, measuring 1.6 mm and with a micropylar apparatus of 27.3 mm in diameter. CONCLUSIONS: The ornamentation of the exochorion presents differences in relation to the tubercles of chorionic cells and the external chorionic reticulum between the eggs of Hg. Leucocelaenus, in comparison with the eggs of Hg. janthinomys and Hg. equinus, and also in relation to those of Aedes aegypti, Ae. albopictus and Ae. bahamensis. In various aspects, the eggs of Hg. leucocelaenus have more resemblance to those of Hg. Equinus than those of Hg. janthinomys, with greater differences presented in relation to the eggs of Hg. spegazzinii and Hg. lucifer.

[Susceptibility of Aedes aegypti larvae to the insecticide temephos in the Federal District, Brazil]. / Suscetibilidade de larvas de Aedes aegypti ao inseticida temefós no Distrito Federal.

OBJECTIVE: To study the susceptibility status of Aedes aegypti to the organophosphate insecticide temephos. METHODS: Samples of Ae. aegypti larvae were obtained, using ovitraps, from eight cities of the Federal District, central Brazil, in 2000 and 2001. Larvae were submitted to the diagnostic dose of 0.012 mg/l temephos, as recommended by standard World Health Organization methodology. Field populations were tested in parallel with reference strains Rockefeller and DIVAL, from the Environmental Surveillance Directory (DIVAL) insectary. The concentration and purity of temephos solutions were verified by gas chromatography. Correlation calculations were performed using StatView - SAS Institute Inc., version 5. Student's t test was used for detecting differences in susceptibility, with significance levels of alpha=0.05. RESULTS: In 2000, Ae. aegypti larvae populations from Taguatinga, Guara, and Nucleo Bandeirante showed resistance to temephos, with mortality ranging from 54.1 to 63.4%. The populations from Gama, Planaltina, and Sobradinho showed altered levels of susceptibility (mortality ranging from 83.6 to 92.8%). The population from Ceilandia was the only susceptible one, with 98% mortality. In 2001, all populations tested were resistant (44.4 to 66.4% mortality). No significant correlation was found between the susceptibility of populations and the distance between the cities of origin, or the amount of insecticide applied in the years preceding the study. CONCLUSIONS: Ae. aegypti susceptibility to temephos is changing in the Federal District. It is essential to continue monitoring the resistance of this vector to insecticides in order to ensure the efficiency of programs aimed at vector control and at the protection of human health.

Aedes albopictus may not be vector of dengue virus in human epidemics in Brazil.

Over 60,500 dengue cases were reported in the state of Espírito Santo (ES), Brazil, between 1995 and 1998. The study's purpose was to identify whether Aedes albopictus was transmitting the dengue virus during an epidemic in the locality of Vila Beth nia (Viana County),Vitória, ES. From April 3 to 9, 1998, blood and serum samples were collected daily for virus isolation and serological testing. Four autochthonous cases were confirmed through DEN 1 virus isolation and two autochthonous cases through MAC ELISA testing. Of 37 Ae. aegypti and 200 Ae. albopictus adult mosquitoes collected and inoculated, DEN1 virus was isolated only from a pool of two Ae. aegypti female mosquitoes. The study results suggest that Ae. albopictus still cannot be considered an inter-human vector in dengue epidemics in Brazil.

A case study of the influence of local weather on Aedes aegypti (L.) aging and mortality.

The survival rate of mosquitoes is an important topic that affects many aspects of decision-making in mosquito management. This study aims to estimate the variability in the survival rate of Ae. aegypti, and climate factors that are related to such variability. It is generally assumed that the daily probability of mosquito survival is independent of natural environment conditions and age. To test this assumption, a three-year fieldwork (2005-2007) and experimental study was conducted at Fortaleza-CE in Brazil with the aim of estimating daily survival rates of the dengue vector Aedes aegypti under natural conditions in an urban city. Survival rates of mosquitoes may be age-dependent and statistical analysis is a sensitive approach for comparing patterns of mosquito survival. We studied whether weather conditions occurring on a particular day influence the mortality observed on that particular day. We therefore focused on the impact of daily meteorological fluctuations around a given climate average, rather than on the influence of climate itself. With regard to survival time, multivariate analyses using the stepwise logistic regression model, adjusted for daily temperature, relative humidity, and saturated vapor pressure deficit (SVPD), suggest that age, the seasonal factor, and the SVPD were the most dependent mortality factors. Similar results were obtained using the Cox proportional hazard model, which explores the relationships between the survival and explanatory variables.

A revision of the genus Mecistostethus Marseul (Histeridae, Histerinae, Exosternini).

We revise the genus Mecistostethus Marseul, sinking the monotypic genus Tarsilister Bruch as a junior synonym. Mecistostethus contains six valid species: Mecistostethus pilifer Marseul, Mecistostethus loretoensis (Bruch), comb. n., Mecistostethus seagorumsp. n., Mecistostethus carltonisp. n., Mecistostethus marseulisp. n., and Mecistostethus flechtmannisp. n. The few existing records show the genus to be widespread in tropical and subtropical South America, from northern Argentina to western Amazonian Ecuador and French Guiana. Only a single host record associates one species with the ant Pachycondyla striata Smith (Formicidae: Ponerinae), but it is possible that related ants host all the species.

A revision of the genus Kaszabister Mazur (Histeridae, Histerinae, Exosternini).

We revise the four species of Kaszabister Mazur, 1972, one of which, Kaszabister barrigaisp. n., is described as new. The other species in the genus are Kaszabister rubellus (Erichson, 1834), Kaszabister ferrugineus (Kirsch, 1873) and Kaszabister carinatus (Lewis, 1888). The species are principally known from the subtropics of South America, with one in Central America. Lectotypes are designated for Kaszabister rubellus and Kaszabister ferrugineus, and a key is provided for all the species. Ants of the genus Solenopsis Westwood, mainly Solenopsis invicta Buren and Solenopsis saevissima (Smith), are documented as hosts of three of the four species.

The influence of local environment on the aging and mortality of Aedes aegypti (L.): case study in Fortaleza-CE, Brazil.

It is generally assumed that the daily probability of survival of mosquitoes is independent of age. To test this assumption we have conducted a three-year experimental fieldwork study (2005-2007) at Fortaleza-CE in Brazil, determining daily survival rates of the dengue vector Aedes aegypti (L.). Survival rates of adult Ae. aegypti may be age-dependent and the statistical analysis is a sensitive approach for comparing patterns of mosquito survival. The mosquito survival data were better fit by a Weibull survival function than by the more traditionally used Gompertz or logistic survival functions. Gompertz, Weibull, or logistic survival functions often fit the survival, and the tails of the survival curves usually appear to fall between the values predicted by the three functions. We corroborate that the mortality of Ae. aegypti in semi-natural conditions may no more be considered as a constant phenomenon during the life of adult mosquitoes but varies according to the age and environmental conditions under a tropical climate. This study estimates the variability in the survival rate of Ae. aegypti and environmental factors that are related to such variability. The statistical analysis shows that the fitting ability, concerning the hazard function, was in decreasing order: Seasonal Cox, the three-parameter Gompertz, and the three-parameter Weibull, that was similar to the three-parameter logistic. The advantage of using the Cox model is that it is convenient for exploring the relationship between survival and several explanatory variables. The Cox model has the advantage of preserving the variable in its original quantitative form and of using a maximum of information. The survival analyses indicate that mosquito mortality is both age- and environment-dependent.

Climate-based models for understanding and forecasting dengue epidemics.

BACKGROUND: Dengue dynamics are driven by complex interactions between human-hosts, mosquito-vectors and viruses that are influenced by environmental and climatic factors. The objectives of this study were to analyze and model the relationships between climate, Aedes aegypti vectors and dengue outbreaks in Noumea (New Caledonia), and to provide an early warning system. METHODOLOGY/PRINCIPAL FINDINGS: Epidemiological and meteorological data were analyzed from 1971 to 2010 in Noumea. Entomological surveillance indices were available from March 2000 to December 2009. During epidemic years, the distribution of dengue cases was highly seasonal. The epidemic peak (March-April) lagged the warmest temperature by 1-2 months and was in phase with maximum precipitations, relative humidity and entomological indices. Significant inter-annual correlations were observed between the risk of outbreak and summertime temperature, precipitations or relative humidity but not ENSO. Climate-based multivariate non-linear models were developed to estimate the yearly risk of dengue outbreak in Noumea. The best explicative meteorological variables were the number of days with maximal temperature exceeding 32°C during January-February-March and the number of days with maximal relative humidity exceeding 95% during January. The best predictive variables were the maximal temperature in December and maximal relative humidity during October-November-December of the previous year. For a probability of dengue outbreak above 65% in leave-one-out cross validation, the explicative model predicted 94% of the epidemic years and 79% of the non epidemic years, and the predictive model 79% and 65%, respectively. CONCLUSIONS/SIGNIFICANCE: The epidemic dynamics of dengue in Noumea were essentially driven by climate during the last forty years. Specific conditions based on maximal temperature and relative humidity thresholds were determinant in outbreaks occurrence. Their persistence was also crucial. An operational model that will enable health authorities to anticipate the outbreak risk was successfully developed. Similar models may be developed to improve dengue management in other countries.