Potential Vectors of Dirofilaria immitis (Leidy, 1856) in Itacoatiara, Oceanic Region of Niterói Municipality, State of Rio de Janeiro, Brazil

Dirofilaria immitis is a widespread mosquito-borne parasite that causes dirofilariasis, a commonly diagnosed disease of dogs that is rarely reported in cats and humans. A mosquito survey was conducted in Itacoatiara in the State of Rio de Janeiro, from March 1995 to February 1996, using canine, feline and human baits. A total of 3,667 mosquitoes were dissected for D. immitis larvae, representing 19 species and 10 genera. From those, Ae. scapularis, Ae taeniorhynchus, Cx. quinquefasciatus, Cx. declarator, Cx. saltanensis and Wy. bourrouli were found infected with D. immitis parasites, and among those, only the first three harbored infective larvae. The majority of larvae were found in the Malpighian tubules (889/936), and larval melanization was observed in the two Aedes species. In descending order, the best vectors were Ae. scapularis, Ae. taeniorhynchus, and Cx. quinquefasciatus which alternate seasonally in importance. Cx. quinquefasciatus is suggested to be a vector to cats. The potential transmission of D. immitis parasites by these three vectors to man is discussed.

Ecology of mosquitoes (Diptera: Culicidae) in areas of Serra do Mar State Park, State of São Paulo, Brazil. I - Monthly frequency and climatic factors

The ecology of mosquitoes were studied (Diptera: Culicidae) in areas of Serra do Mar State Park, State of São Paulo, Brazil. Systematized monthly human bait collections were made three times a day, for periods of 2 or 3 h each, in sylvatic and rural areas for 24 consecutive months (January 1991 to December 1992). A total of 24,943 specimens of adult mosquitoes belonging to 57 species were collected during 622 collective periods. Coquillettidia chrysonotum was the most frequent collected mosquito (45.8%) followed by Aedes serratus (6.8%), Cq. venezuelensis (6.5%), Psorophora ferox (5.2) and Ps. albipes (3.1%). The monthly averages of temperature and relative humidity were inserted in the ten-year average limits of maximum and minimum of the previous ten-years. Rainfall accompanied the curve of the ten-year averages. Those climatic factors were influential in the incidence of some species; temperature: Anopheles cruzii, An. mediopunctatus, Ae. scapularis, Ae. fulvus, Cq. chrysonotum, Cq. venezuelensis, Runchomyia reversa, Wyeomyia dyari, Wy. confusa, Wy. shannoni, Wy. theobaldi and Limatus flavisetosus; relative humidity: Ae. serratus, Ae. scapularis, Cq. venezuelensis and Ru. reversa; rainfall: An. cruzii, Ae. scapularis, Ae. fulvus, Cq. venezuelensis Ru. reversa, Wy. theobaldi and Li. flavisetosus.

Ecology of mosquitoes (Diptera: Culicidae) in areas of Serra do Mar State Park, State of São Paulo, Brazil. II - Habitat distribution

The mosquito (Diptera: Culicidae) ecology was studied in areas of Serra do Mar State Park, State of São Paulo, Brazil. Systematized biweekly human bait collections were made three times a day, for periods of 2 or 3 h each, in sylvatic and rural areas for 24 consecutive months (January 1991 to December 1992). A total of 24,943 adult mosquitoes belonging to 57 species were collected during 622 collective periods. Aedes scapularis, Coquillettidia chrysonotum, Cq. venezuelensis, Wyeomyia dyari, Wy. longirostris, Wy. theobaldi and Wy. palmata were more frequently collected at swampy and at flooded areas. Anopheles mediopunctatus, Culex nigripalpus, Ae. serratus, Ae. fulvus, Psorophora ferox, Ps. albipes and the Sabethini in general, were captured almost exclusively in forested areas. An. cruzii, An. oswaldoi and An. fluminensis were captured more frequently in a residence area. However, Cx. quinquefasciatus was the only one truly eusynanthropic. An. cruzii and Ae. scapularis were captured feeding on blood inside and around the residence, indicating that both species, malaria and arbovirus vectors respectively, may be involved in the transmission of these such diseases in rural areas.

Comparison of dengue infection in human mononuclear leukocytes with mosquito C6/36 and mammalian Vero cells using flow cytometry to detect virus antigen

Fluorescent activated cell sorter (FACS) analysis is useful for the detection of cellular surface antigens and intracellular proteins. We used this methodology in order to detect and quantify dengue antigens in highly susceptible cells such as clone C6/36 (Aedes albopictus) and Vero cells (green monkey kidney). Additionally, we analyzed the infection in vitro of human peripheral blood mononuclear leukocytes (PBML). FACS analysis turned out to be a reliable technique to quantify virus growth in traditional cell cultures of C6/36 as well as Vero cells. High rates of infection were achieved with a good statistical correlation between the virus amount used in infection and the percentage of dengue antigen containing cells detected in infected cultures. We also showed that human monocytes (CD14+) are preferred target cells for in vitro dengue infection among PBML. Monocytes were much less susceptible to virus infection than cell lines but they displayed dengue antigens detected by FACS five days after infection. In contrast, lymphocytes showed no differences in their profile for dengue specific immunofluorescence. Without an animal model to reproduce dengue disease, alternative assays have been sought to correlate viral virulence with clinical manifestations and disease severity. Study of in vitro interaction of virus and host cells may highlight this relationship.

Ecology of mosquitoes (Diptera: Culicidae) in areas of Serra do Mar State Park, State of São Paulo, Brazil. III - daily biting rhythms and lunar cycle influence

The ecology of mosquito species (Diptera: Culicidae) was studied in areas of the Serra do Mar State Park, State of São Paulo, Brazil. The influence of the lunar cycle and the daily biting rhythms of mosquito populations were analyzed. Systematized biweekly human bait collections were made in a silvatic environment for 24 consecutive months (January 1991 to December 1992). A total of 20,591 specimens of adult mosquitoes belonging to 55 species were collected from 545 catches. Sabethini species were captured exclusively during daylight periods, with the exception of Trichoprosopon digitatum, while members of Anophelinae predominated during nocturnal hours. Members of the subfamily Culicinae that were collected primary during nocturnal periods included Culex nigripalpus, Coquillettidia chrysonotum and Cq. venezuelensis while daytime catches included Psorophora ferox and Ps. albipes. Others members of culicines mosquitoes that were collected during both day and night included: Aedes serratus, Ae. scapularis and Ae. fulvus. Lunar cycles did not appear to influence the daily biting rhythms of most mosquito species in the area, but larger numbers of mosquitoes were collected during the new moon. Ae. scapularis were captured mainly during the full moon.

Vector competence of Culex quinquefasciatus say from different regions of Brazil to Dirofilaria immitis

The vector competence of Culex quinquefasciatus from five localities in Brazil to Dirofilaria immitis was evaluated experimentally. Females from each locality were fed on an infected dog (~ 6 microfilariae/µl blood). A sample of blood fed mosquitoes were dissected approximately 1 h after blood meal. These results demonstrated that all had ingested microfilariae (mean, 4.8 to 24.6 microfilariae/mosquito). Fifteen days after the infected blood meal, the infection and infective rates were low in all populations of Cx. quinquefasciatus. The mean number of infective larvae detected in the head and proboscis of these mosquitoes was 1-1.5. The vector efficiency, the number of microfilariae ingested/number of infective larvae, was low for all populations of Cx. quinquefasciatus. However, the survival rate for all populations was high (range 50-75%). The survival rate of Aedes aegypti assayed simultaneously for comparison was low (24.7%), while the vector efficiency was much higher than for Cx. quinquefasciatus. These data suggest that the vector competence of all assayed populations of Cx. quinquefasciatus to D. immitis in Brazil is similar and that this species is a secondary vector due to its low susceptibility. Nevertheless, vector capacity may vary between populations due to differences in biting frequency on dogs that has been reported in Brazil.

Dengue situation in Brazil by year 2000

Dengue virus types 1 and 2 have been isolated in Brazil by the Department of Virology, Instituto Oswaldo Cruz, in 1986 and 1990 respectively, after many decades of absence. A successful continental Aedes aegypti control program in the Americas, has been able to eradicate the vector in most countries in the 60's, but the program could not be sustained along the years. Dengue viruses were reintroduced in the American region and the infection became endemic in Brazil, like in most Central and SouthAmerican countries and in the Caribbean region, due to the weaning of the vector control programs in these countries. High demographic densities and poor housing conditions in large urban communities, made the ideal conditions for vector spreading. All four dengue types are circulating in the continent and there is a high risk of the introduction in the country of the other two dengue types in Brazil, with the development of large epidemics. After the Cuban episode in 1981, when by the first time a large epidemic of dengue hemorrhagic fever and dengue shock syndrome have been described in the Americas, both clinical presentations are observed, specially in the countries like Brazil, with circulation of more than one dengue virus type. A tetravalent potent vaccine seems to be the only possible way to control the disease in the future, besides rapid clinical and laboratory diagnosis, in order to offer supportive treatment to the more severe clinical infections.

Yellow fever

With the infestation by Aedes aegypti, urban yellow fever might already exist. This did not occur because of either the lacking of a sufficient contact between the diseased individual and the A. aegypti or perhaps because this, after sixty years without transmitting the virus, needs an adaptation phase to infecting again.

The use of bacterial larvicides in mosquito and black fly control programmes in Brazil

Bacillus spp. based larvides are increasingly replacing, with numerous advantages, chemical insecticides in programmes for controlling black fly and mosquito populations. Brazil was among the pioneers in adopting Bacillus thuringiensis israelensis (B.t.i) to control black flies. However, the major current mosquito control programme in Brazil, the Programme for Eradication of Aedes aegypti launched in 1997, only recently decided to replace temephos by B.t.i based larvicides, in the State of Rio de Janeiro. In the last decade, works developed by research groups in Brazilian institutions have generated a significant contribution to this subject through the isolation of B. sphaericus new strains, the development of new products and the implementation of field trials of Bacillus efficacy against mosquito species under different environmental conditions.

Expression of mosquito active toxin genes by a Colombian native strain of the gram-negative bacterium Asticcacaulis excentricus

Mosquito control with biological insecticides, such as Bacillus sp. toxins, has been used widely in many countries. However, rapid sedimentation away from the mosquito larvae feeding zone causes a low residual effect. In order to overcome this problem, it has been proposed to clone the Bacillus toxin genes in aquatic bacteria which are able to live in the upper part of the water column. Two strains of Asticcacaulis excentricus were chosen to introduce the B. sphaericus binary toxin gene and B. thuringiensis subsp. medellin cry11Bb gene cloned in suitable vectors. In feeding experiments with these aquatic bacteria, it was shown that Culex quinquefasciatus, Aedes aegypti, and Anopheles albimanus larvae were able to survive on a diet based on this wild bacterium. A. excentricus recombinant strains were able to express both genes, but the recombinant strain expressing the B. sphaericus binary toxin was toxic to mosquito larvae. Crude protease A. excentricus extracts did not degrade the Cry11Bb toxin. The flotability studies indicated that the recombinant A. excentricus strains remained in the upper part of the water column longer than the wild type Bacillus strains.

Identification of entomopathogenic Bacillus isolated from Simulium (Diptera, Simuliidae) larvae and adults

Entomopathogenic bacteria isolated from Simulium larvae and adults from breeding sites in the states of São Paulo and Rio de Janeiro, Brazil, were identified as 18 strains of Bacillus thuringiensis and one of B. sphaericus. Most of these strains were serotyped according to their flagellar antigens. However, nine of the B. thuringiensis samples, could not be serotyped and were designated as "autoagglutinating"; they were also shown to be toxic in preliminary tests against Aedes aegypti larvae. Additionally, B. sphaericus was also shown to be toxic towards Culex quinquefasciatus larvae.

Ochlerotatus albifasciatus in rain pools of Buenos Aires: seasonal dynamics and relation to environmental variables

The immature stages of Ochlerotatus albifasciatus develop in temporary pools. The present study aims at evaluating the seasonal dynamics of the aquatic stages of this mosquito, also analyzing the relationship among their presence and breeding success to some relevant climatic and environmental variables in the ephemeral rain pools of an urban park. Nineteen cohorts of O. albifasciatus that developed synchronously after rain events were recorded in all seasons. The proportions of mosquito-positive pools were significantly higher during the fall-winter period than in the spring-summer months (p < 0.001). The presence of this mosquito species was positively related to the amount of rain (p < 0.001), whereas negatively correlated to air temperature (p < 0.05) within a 5.2 to 29.7ºC range. The distribution of the number of cohorts per pool throughout the year was grouped (variance/mean: 3.96), indicating that these habitats were not equally suitable as breeding sites. The immature stages of O. albifasciatus were detected in pools belonging to all of the categories of surface area, depth, duration, vegetation cover, and insolation. However, the proportion of pools where immature mosquitoes were detected was positively and significantly related to surface, depth, duration, and vegetation cover. On the other hand, the proportion of mosquito-positive pools was higher at an intermediate insolation degree. Our results suggest that although preimaginal stages were present in all seasons, high temperatures may be unfavorable to larval development, and substrate vegetation may regulate water temperature. The positive relationship between the proportion of mosquito-positive pools and pool size and duration might reflect a strategy of O. albifasciatus to accomplish immature development.

Laboratory evaluation of Mesocyclops annulatus (Wierzejski, 1892) (Copepoda: Cyclopidea) as a predator of container-breeding mosquitoes in Argentina

In laboratory bioassays we tested the predatory capacity of the copepod Mesocyclops annulatus on Aedes aegypti and Culex pipiens larvae. A single adult female of M. annulatus caused 51.6% and 52.3% mortality of 50 first instar larvae of Ae. aegypti and Cx. pipiens respectively, in a 72 h test period. When alternative food was added to the containers, mortality rates declined to 16% and 10.3% for Ae. aegypti and Cx. pipiens respectively. When 50 first instar larvae of each of the two mosquito species tested were placed together with a single adult female of M. annulatus, mortality rates were 75.5% for Ae. aegypti larvae and 23.5% for Cx. pipiens larvae in a three day test period. Different density of adult females of M. annulatus ranged from 5 to 25 females produced mortality rates of Ae. aegypti first instar larvae from 50% to 100% respectively. When a single adult female of M. annulatus was exposed to an increasing number of first-instar Ae. aegypti larvae ranging from 10 to 100, 100% mortality was recorded from 1 to 25 larvae, then mortality declined to 30% with 100 larvae. The average larvae killed per 24 h period by a single copepod were 29.

Mortality in immatures of the floodwater mosquito Ochlerotatus albifasciatus (Diptera: Culicidae) and effects of parasitism by Strelkovimermis spiculatus (Nematoda: Mermithidae) in Buenos Aires Province, Argentina

Life tables were constructed for six cohorts of immature stages of the floodwater mosquito Ochlerotatus albifasciatus (Macquart) in a park in Buenos Aires, highlighting the mortality attributable to the parasitic nematode, Strelkovimermis spiculatus Poinar & Camino. Two cohorts were selected to compare parasite incidence in all mosquito stages when low and high parasitism occurred. Development time of Oc. albifasciatus from first instar to adult was 7.7-10 days in the spring, 6 days in the summer, and 10.9-21.9 days in the fall. Survival was estimated as 0-1.4% in the spring, 2% in the summer and 0.2-4.4% in the fall. The highest "K" value (Killing power) occurred during a fall cohort when prevalence of the parasite was 86.9%, and the lowest in a spring cohort. Parasitism occurred during all seasons, but S. spiculatus persisted to adult only in the summer and fall, when adult mosquitoes developed from parasitized third and fourth instars larvae. The abundance of S. spiculatus differed between old and young larvae only when parasite prevalence was the highest. Although pupae and adults of Oc. albifasciatus were parasitized, no pupal mortality attributable to parasitism was recorded. The proportion of parasitized adults ranged from 14.2% and 5.7% in the two cohorts compared. Pupal wet weight and adult wing lengths did not differ between parasitized and unparasitized individuals.