Mosquito isolates of Ross River virus from Cairns, Queensland, Australia

During 1996-1998 60,619 mosquitoes were collected around Cairns, Australia and processed for Alphavirus isolation. Thirty-three isolates of Ross River (RR) virus were made from 9 species, Aedes imprimens, Aedes kochi, Aedes notoscriptus, Aedes vigilax, Culex annulirostris, Culex gelidus, Mansonia septempunctata, Verrallina (formerly Aedes) carmenti, and Verrallina lineatus. Attempts to isolate RR virus from 121 Aedes aegypti were unsuccessful. Twenty six (79%) of the isolates came from within 1 km of a colony of spectacled flying-foxes, Pteropus conspicillatus. The minimum infection rate for these mosquitoes was 1.0 compared with 0.2 per 1,000 for mosquitoes trapped at all other sites. Ross River virus has not previously been isolated from Ae. imprimens, Cx. gelidus, Ma. septempunctata, Ve. carmenti, or Ve. lineatus. This is also the first isolation of an arbovirus from Cx. gelidus in Australia. In conclusion, the vector status of Ve. carmenti, Ae. aegypti and Mn. septempunctata warrants further study. This study also provides evidence that P. conspicillatus may be a reservoir host.

Point source inoculation of Mesocyclops (Copepoda : Cyclopidae) gives widespread control of Ochlerotatus and Aedes (Diptera : Culicidae) immatures in service manholes and pits in North Queensland, Australia

This study details the novel application of predacious copepods, genus Mesocyclops, for control of Ochlerotatus tremulus (Theobald) group and Aedes aegypti (L.) mosquito larvae in subterranean habitats in north Queensland, Australia. During June 1997, 50 Mesocyclops sp. I were inoculated into one service manhole in South Townsville. Wet season rainfall and flooding in both 1998 and 2000 was responsible for the dispersal of copepods via the underground pipe system to 29 of 35 manholes over an area of 1.33 km(2). Significant reductions in Aedes and Ochlerotatus larvae ensued. In these habitats, Mesocyclops and Metacyclops were able to survive dry periods, when substrate moisture content ranged from 13.8 to 79.9%. At the semiarid inland towns of Hughenden and Richmond, cracking clay soil prevents drainage of water from shallow service pits where Oc. tremulus immatures numbered from 292-18,460 per pit. Introduction of Mesocyclops copepods into these sites during May 1999 resulted in 100% control of Oc. tremulus for 18 mo. One uninoculated pit subsequently became positive for Mesocyclops with resultant control of mosquito larvae.

Flavivirus isolations from mosquitoes collected from western Cape York Peninsula, Australia, 1999-2000

After the 1st appearance of Japanese encephalitis virus (JE) on mainland Australia in 1998, a study was undertaken to investigate whether JE had become established in enzootic transmission cycles on western Cape York Peninsula. Adult mosquitoes were collected during the late wet season from Kowanyama and Pormpuraaw in April 1999, and Pormpuraaw and Barr's Yard in April 2000. Despite processing 269,270 mosquitoes for virus isolation, no isolates of JE were obtained. However, other flaviviruses comprising Murray Valley encephalitis virus, Kunjin virus, Alfuy virus, and Kokobera virus (KOK) were isolated. Isolates of the alphaviruses Ross River virus, Barmah Forest virus (BF), and Sindbis virus (SIN) also were obtained. The majority (88%) of isolates were from members of the Culex sitiens subgroup. Single isolates of KOK, BF and SIN were obtained from Ochlerotatus vigilax, Oc. normanensis, and Anopheles bancroftii, respectively. The isolations of flaviviruses during the late wet season indicate that conditions were suitable for flavivirus activity in the area. No evidence was found to suggest that JE has become established in enzootic transmission cycles on western Cape York, although study sites and field trips were limited.

Studies on mosquitoes (Diptera: Culicidae) and anthropic environment: 8- Survey of adult behaviour of Spissipes Section species of Culex (Melanoconion) in South-Eastern Brazil

A study of Spissipes Section of Culex (Melanoconion) adults behaviour was carried out from August 1992 through December 1993 in human dominated (anthropic) environment in the Ribeira Valley, S.Paulo State, Brazil. By sampling at several sites it the dominance of Culex ribeirensis and Cx. sacchettae became evident even through a total number of ten species was recorded. Those two mosquitoes showed a clear tendency to frequent the domiciliary environment where they were caught, both indoor and outdoor, through the use of the human bait. In the outside environments, the residual patchy forests seems to display a concentration role, from which these adults spread to the open land and reach the dwellings. As their vector competence has been demonstrated through the virus isolations in natural conditions, it is advisable to pay attention to the presence of these mosquitoes in the man-made environment.

Comparison of feeding and defecation patterns between fifth-instar nymphs of Triatoma patagonica (Del Ponte, 1929) and Triatoma infestans (Klug, 1934) under laboratory conditions

Among the vectors of Chagas disease, Triatoma patagonica is a species in the process of adaptation to the human environment. However, its vector competence is not well known. This study had the aim of evaluating and comparing feeding and defecation patterns among fifth-instar nymphs of Triatoma patagonica and Triatoma infestans that were fed ad libitum. The results showed that nymphs of Triatoma patagonica had a feeding pattern similar to that of Triatoma infestans. Sixty nine percent and 58% of nymphs of Triatoma patagonica and Triatoma infestans, respectively, produced their first defecation within five minutes after being fed. Triatoma patagonica defecated during feeding, with an average time until first defecation that was shorter than that of Triatoma infestans (3.4 and 6.2 min, respectively). The nymphs of Triatoma patagonica were capable of defecating during or immediately after feeding.

Twenty-eight years of Aedes albopictus in Brazil: a rationale to maintain active entomological and epidemiological surveillance

INTRODUCTION: Aedes albopictus was first detected in Brazil in 1986. This mosquito species presents a major threat to public health because Brazilian populations have shown substantial vector competence for arboviruses such as dengue and chikungunya. METHODS: We updated the records of Ae. albopictus in several States of Brazil, focusing on areas in which its presence had been reported after 2002. RESULTS: Twenty-eight years after its arrival in Brazil, Ae. albopictus has been detected in 24 of 27 States. CONCLUSIONS: The rapid spread of this species and its high vector competence demonstrate the danger of Ae. albopictus in Brazil.

Malpighian tubule polytene chromosomes of Culex quinquefasciatus (Diptera, Culicinae)

Dipteran polytene chromosomes provide an excellent model for understanding in species complexes, as well as for structural and functional cytogenetics. The status of species in the Culex pipiens complex is controversial and the use of polytene chromosomes for cytogenetic analysis in the subfamily Culicinae has been difficult because of methodological problems. In this study, Malpighian tubule polytene chromosomes were obtained from young (0 to 12 h, 20ºC) and old (20 to 42 h, 28ºC) laboratory-bred C. pipiens quinquefasciatus pupae. The chromosome maps for this species were constructed and compared with published data for C. pipiens pipiens and C. p. quinquefasciatus. Although the banding patterns were conserved between subspecies, analysis of the structural variations in the bands and interbands revealed differences apparently related to the physiological stage and ecogeographical strain. The organization of the centromeric regions in larval and pupal chromosomes showed greater similarity to each other than did those of pupal and adult chromosomes. The use of pupal polytene chromosomes for in situ hybridization with vector competence probes is discussed.

A technique for preparing polytene chromosomes from Aedes aegypti (Diptera, Culicinae)

Polytene chromosome preparations were obtained from larval, pupal and adult female Malpighian tubules of Aedes aegypti. The Malpighian tubules of the pupae (0-4 h old) from larvae reared at 20ºC provided the best cytogenetic analysis. The interaction of nucleic acids and proteins that influence the spreading of the chromosomes could be reduced with the preparation technique of the sheets submitted to a stronger treatment starting with the hypotony of tissue and successive bathings with acetic acid. A simple technique should facilitate molecular cytogenetics used in the location of resistance and vector competence genes.

Dengue virus detection by using reverse transcription-polymerase chain reaction in saliva and progeny of experimentally infected Aedes albopictus from Brazil

Oral susceptibility and vertical transmission of dengue virus type 2 (DENV-2) in an Aedes albopictus sample from Rio de Janeiro was estimated. The infection (36.7%) and transmission (83.3%) rates for Ae. albopictus were higher than those of an Ae. aegypti colony used as control, 32.8 and 60%, respectively. Fourth instar larvae and females descendants of 48.5 and 39.1% of experimentally infected Ae. albopictus showed to harbor the virus. The oral susceptibility and the high capacity to assure vertical transmission exhibited by Ae. albopictus from Brazil reinforce that this species may play a role in the maintenance of the virus in nature and be a threat for dengue control in the country.

Preliminary data on the performance of Aedes aegypti and Aedes albopictus immatures developing in water-filled tires in Rio de Janeiro

A monthly survey of Aedes aegypti and Aedes albopictus immatures in discarded tires at a site in metropolitan Rio de Janeiro showed that Ae. albopictus was much more abundant in the rainy season, but Ae. aegypti abundance showed a less clear seasonal pattern. Pupal masses for Ae. albopictus showed a seasonal trend. In contrast, Ae. aegypti pupae did not show any clear trend in weight. Large Ae. albopictus pupae were found in the warmer months, when water volume was higher, pH lower, and larval abundance lower. Further studies should be carried out to assess how seasonal variations in body size may impact vector competence of these species in Brazil.

Mosquito (Diptera: Culicidae) Diversity of a Forest-Fragment Mosaic in the Amazon Rain Forest

To study the impact of Amazonian forest fragmentation on the mosquito fauna, an inventory of Culicidae was conducted in the upland forest research areas of the Biological Dynamics of Forest Fragments Project located 60 km north of Manaus, Amazonas, Brazil. The culicid community was sampled monthly between February 2002 and May 2003. CDC light traps, flight interception traps, manual aspiration, and net sweeping were used to capture adult specimens along the edges and within forest fragments of different sizes (1, 10, and 100 ha), in second-growth areas surrounding the fragments and around camps. We collected 5,204 specimens, distributed in 18 genera and 160 species level taxa. A list of mosquito taxa is presented with 145 species found in the survey, including seven new records for Brazil, 16 new records for the state of Amazonas, along with the 15 morphotypes that probably represent undescribed species. No exotic species [Aedes aegypti (L.) and Aedes albopictus (Skuse)] were found within the sampled areas. Several species collected are potential vectors of Plasmodium causing human malaria and of various arboviruses. The epidemiological and ecological implications of mosquito species found are discussed, and the results are compared with other mosquito inventories from the Amazon region.

Density and survival rate of Culex quinquefasciatus at Parque Ecologico do Tiete, Sao Paulo, Brazil

Parity rate, gonotrophic cycle length, and density of a Culex quinquefasciatus female population was estimated at the Parque Ecologico do Tiete (PET), Sao Paulo, Brazil. Adult Cx. quinquefasciatus females were collected from vegetation along the edges of a polluted drainage canal with the use of a battery-powered backpack aspirator from September to November 2005 and from February to April 2006. We examined 255 Cx. quinquefasciatus ovaries to establish the parity rate of 0.22 and determined the gonotrophic cycle length under laboratory conditions to be 3 and 4 days. From these data, we calculated the Cx. quinquefasciatus survival rate to be 0.60 and 0.68 per day. Density of the Cx. quinquefasciatus female (5.71 females per m(2)) was estimated based on a population size of 28,810 individuals divided by the sampled area of 5,040 m(2). Results of all experiments indicate medium survivorship and high density of the Cx. quinquefasciatus female population. This species is epidemiologically relevant in the PET area and should be a target of the vector control program of Sao Paulo municipality.

TICK INNATE IMMUNITY

Ticks are blood feeding parasites transmitting a wide variety of pathogens to their vertebrate hosts. The vector competence of ticks is tightly linked with their immune system. Despite its importance, our knowledge of tick innate immunity is still inadequate and the limited number of sufficiently characterized immune molecules and cellular reactions are dispersed across numerous tick species. The phagocytosis of microbes by tick hemocytes seems to be coupled with a primitive complement-like system, which possibly involves self/nonself recognition by fibrinogen-related lectins and the action of thioester-containing proteins. Ticks do not seem to possess a pro-phenoloxidase system leading to melanization and also coagulation of tick hemolymph has not been experimentally proven. They are capable of defending themselves against microbial infection with a variety of antimicrobial peptides comprising lysozymes, defensins and molecules not found in other invertebrates. Virtually nothing is known about the signaling cascades involved in the regulation of tick antimicrobial immune responses. Midgut immunity is apparently the decisive factor of tick vector competence. The gut content is a hostile environment for ingested microbes, which is mainly due to the antimicrobial activity of hemoglobin fragments generated by the digestion of the host blood as well as other antimicrobial peptides. Reactive oxygen species possibly also play an important role in the tick-pathogen interaction. The recent release of the Ixodes scapularis genome and the feasibility of RNA interference in ticks promise imminent and substantial progress in tick innate immunity research.

Susceptibility of mosquito vectors to Dirofilaria immitis on Madeira Island, Portugal

Dirofilaria immitis (Leidy, 1856), an agent of heartworm disease, is an important parasite from both the veterinary standpoint and as a model to study human filariasis. It is a mosquito-borne filarial nematode which inhabits the right ventricle and pulmonary arteries of dogs. D. immitis is an important disease agent on Madeira Island with about 30% of dogs testing positive for this worm. Nevertheless, the vectors of this parasite in Madeira have never been studied, nor has the interaction between pathogen and vector, or the environmental variables that might influence heartworm transmission. Innate susceptibility to infection is only one component of vector competence, and field isolation of naturally infected mosquitoes has shown the capability of D. immitis to exploit a great diversity of vector species under natural conditions. The purpose of this work was to determine which mosquitoes are vectors of heartworm disease, the relation between population density and environment, and the association between immune response of the vector to the filarial parasite. Seasonal abundance of Culex theileri and Culex pipiens molestus was studied. Correlation and canonical correspondence analysis were performed using abundance data of these two species with selected weather variables, including mean temperature, relative humidity and accumulated precipitation. The most important factor determining Cx. theileri abundance was accumulated precipitation, while Cx. pipiens molestus abundance did not have any relationship with weather variables. Field studies were performed to verify whether Cx. theileri Theobald functions as a natural vector of D. immitis on Madeira Island, Portugal. Cx. theileri tested positive for D. immitis for the first time. The same study was made regarding Cx. p. molestus. Two abnormal L2 stage filarial worms were found in Malpighian tubules in field caught Cx. p. molestus. In the laboratory, two strains of Cx. p. molestus were studied for their susceptibility to D. immitis. None presented infective-stage larvae. Finally, because Cx. p. molestus is an autogenous mosquito, we evaluated the reproductive costs when this mosquito mounts an immune response against D. immitis in the absence of a blood meal. This mosquito showed an active immune response when inoculated intrathoracically with microfilariae (mf) of the heartworm. The ovaries from mosquitoes undergoing melanotic encapsulation developed more eggs than those which could not melanize the mf. This fact is contradictory with some previous studies of reproductive costs in Armigeres subalbatus and Ochlerotatus trivittatus, and it was the first time that an autogenous mosquito was used to study this subject.

Comportamento adaptativo de Lutzomyia longipalpis (Diptera: Psychodidae) às áreas urbanas da cidade de Teresina-PI.

The sand fly Lutzomyia longipalpis (Diptera: Psychodidae) is currently appointed as the main vector of visceral leishmaniasis in the Americas. The growth of cities in areas originally endemics to American Visceral Leishmaniasis (AVL) resulted in the spread of the disease at the same time that observed the adaptation of this species to the urban environment.Changes in behavior of L.longipalpis that enabled the adapt to increasing losings of biodiversity, as well as the frequent exposure of the vector to insecticides evident in urban areas, could justify the increasing population of the species and consequently the spread of disease for these environments .Thus, we selected sixty houses spread among three areas with increasing stages of occupation of an area endemic for AVL in Teresina-PI. We evaluated the correlation between the density of L.longipalpis captured and different aspects, such as population density of animals, vegetation cover and socio-economic aspects in each house. In addition to the correlations, the feeding preference of the vector between the predominant plant species in the neighborhoods, as well as the presence of metabolic mechanisms of resistance among the captured insects were tested. The results showed that over the growing occupations, represented by three areas, L.longipalpis demonstrate its adaptive nature through an apparent opportunistic behavior in relation to sources of carbohydrates and blood. On the evolutionary point of view, this behavior may have favored its vector competence in urban areas among the limited presence of food sources, as well as in various environments encountered.