Ana Caroline Dalla Bona

Paridade de Anopheles cruzii em Floresta Ombrófila Densa no Sul do Brasil

OBJECTIVE To determine the parity and ovarian development of Anopheles cruzii species during the seasons. METHODS Collections were carried out fortnightly in the morning in the Palmito State Park in the municipality of Paranaguá, Southern Brazil, between April 2004 and April 2005. Adult mosquitoes were captured using human landing rate. Dissections were performed using Detinovas and Polovodovas methods and follicular development was assessed following Christophers and Mers criteria. RESULTS A total of 208 specimens of Anopheles cruzii were dissected. Most females dissected were nulliparous in the seasons; 14.4% of them were found to be nulliparous above Christophers and Mers stage II, which shows previous blood meal prior to the first oviposition. It was observed that Anopheles cruzii populations comprised young mosquitoes, probably due to high mortality among parous females. CONCLUSIONS The likely gonotrophic discordance is epidemiologically relevant because female mosquitoes can search for more than one host to complete the maturation of their eggs.

Insecticide resistance and genetic variability in natural populations of Aedes (Stegomyia) aegypti (Diptera: Culicidae) from Colombia

Mosquito control prevails as the most efficient method to protect humans from the dengue virus, despite recent efforts to find a vaccine for this disease. We evaluated insecticide resistance and genetic variability in natural populations of Aedes aegypti (Linnaeus, 1762) from Colombia. This is the first Colombian study examining kdr mutations and population structure. Bioassays with larvae of three mosquito populations (Armenia, Calarca and Montenegro) were performed according to the World Health Organization (WHO) guidelines, using Temephos. For the analysis of the Val1016Ile mutation and genetic diversity, we sampled recently-emerged adults from four mosquito populations (Armenia, Calarca, Montenegro and Barcelona). Following the WHO protocol, bioassays implemented with larvae showed resistance to Temephos in mosquito populations from Armenia (77% ± 2) and Calarca (62% ± 14), and an incipient altered susceptibility at Montenegro (88% ± 8). The RR95 of mosquito populations ranged from 3.7 (Montenegro) to 6.0 (Calarca). The Val1016Ile mutation analysis of 107 genotyped samples indicates that 94% of the specimens were homozygous for the wild allele (1016Val) and 6% were heterozygous (Val1016Ile). The 1016Ile allele was not found in Barcelona. Genetic variability analysis found three mitochondrial lineages with low genetic diversity and gene flow. In comparison with haplotypes from the American continent, those from this study suggest connections with Mexican and North American populations. These results confirm that a continuous monitoring and managing program of A. aegypti resistance in the state of Quindio is required.

Genetic profile and molecular resistance of Aedes (Stegomyia) aegypti (Diptera: Culicidae) in Foz do Iguaçu (Brazil), at the border with Argentina and Paraguay

The genetic variability of populations of Aedes (Stegomyia) aegypti (Linnaeus, 1762) is often low due to the intense selection pressure caused by chemical control measures. In this study, we evaluated the susceptibility of larvae and adults of this mosquito to chemical insecticides, the frequency of the Val1016IIe mutation, and the genetic variability of the mitochondrial ND4 gene fragment in the urban area of Foz do Iguacu, Parana. The populations of A. aegypti in the southern and central regions of the city were resistant to the diagnostic dose of temephos 0.0162 ppm. Additionally, we detected difference in susceptibility in the northern population. The resistance ratios (RR95) were 3.8, 4.1, and 4.6 for the populations in the north, central, and south of the city, respectively. The entire population of A. aegypti in Foz do Iguacu is resistant to pyrethroids. The mortality rates are as follows: 72.2% for cypermethrin (CD: 146 mg i.a./m2) and 57% for deltamethrin (CD: 18 mg i.a./m2). These results corroborate the hypothesis that the mutated 1016Ile allele is present in all analyzed strata. Out of the 234 samples genotyped for the Val1016Ile mutation, 15% were homozygous dominant for the wild allele (Val/Val), 62% were heterozygous (Va/Ile) and 23% were homozygous for the recessive mutation (Ile/Ile). With respect to the genetic variability of the mitochondrial ND4 gene fragment, 93% of the sequences analyzed belonged to haplotype 1, and 7% belonged to haplotype 2. The genetic diversity was low, the fixation index was not significant, and gene flow was high. The control of A. aegypti in Foz do Iguacu using temephos and pyrethroids may be compromised because of the reduced vector susceptibility. Populations of A. aegypti, that undergoes a sudden reduction in effective population size and become resistant to pyrethroids may differ from the original population in vector capacity.

Crepuscular activity of culicids (Diptera, Culicidae) in the peridomicile and in the remaining riparian forest in Tibagi river, State of Paraná, Brazil

Crepuscular activity of culicids (Diptera, Culicidae) in the peridomicile and in the remaining riparian forest in Tibagi river, State of Parana, Brazil. Human-attracted mosquitoes were collected for one hour, around sunset time (half hour before and half after), from April to December 2006, in two environments (riparian forest and near houses), in Tibagi river basin, Palmeira municipality, State of Parana. Seven-hundred forty-nine mosquitoes, belonging to 13 species, were collected. Psorophora champerico Dyar & Knab, 1906 (42.86%) and Psorophora discrucians (Walker, 1856) (40.59%) were the most frequent species. No significant differences between quantities of Ps. champerico (t = -0.792; d.f. = 16; p = 0.43) and Ps. discrucians (t = 0.689; d.f. = 16; p = 0.49) obtained in riparian forest and near houses were observed, indicating similar conditions for crepuscular activity of these species in both environments. Psorophora champerico and Ps. discrucians responded (haematophagic activity) to environmental stimuli associated with the twilight hours differently in distinct habitats studied. The former species is registered for the first time in the Atlantic forest biome.

Genetic diversity of dengue virus serotypes 1 and 2 in the State of Paraná, Brazil, based on a fragment of the capsid/premembrane junction region

INTRODUCTION The precise identification of the genetic variants of the dengue virus is important to understand its dispersion and virulence patterns and to identify the strains responsible for epidemic outbreaks. This study investigated the genetic variants of the capsid-premembrane junction region fragment in the dengue virus serotypes 1 and 2 (DENV1-2). METHODS Samples from 11 municipalities in the State of Paraná, Brazil, were provided by the Central Laboratory of Paraná. They were isolated from the cell culture line C6/36 (Aedes albopictus) and were positive for indirect immunofluorescence. Ribonucleic acid (RNA) extracted from these samples was submitted to the reverse transcription polymerase chain reaction (RT-PCR) and nested PCR. RESULTS RT-PCR revealed that 4 of the samples were co-infected with both serotypes. The isolated DENV-1 sequences were 95-100% similar to the sequences of other serotype 1 strains deposited in GenBank. Similarly, the isolated DENV-2 sequences were 98-100% similar to other serotype 2 sequences in GenBank. According to our neighbor-joining tree, all strains obtained in this study belonged to genotype V of DENV-1. The DENV-2 strains, by contrast, belonged to the American/Asian genotypes. CONCLUSIONS The monitoring of circulating strains is an important tool to detect the migration of virus subtypes involved in dengue epidemics.

Evaluation of arboviruses of public health interest in free-living non-human primates (Alouatta spp., Callithrix spp., Sapajus spp.) in Brazil

INTRODUCTION The aim of the present study was to evaluate the presence of arboviruses from the Flavivirus genus in asymptomatic free-living non-human primates (NHPs) living in close contact with humans and vectors in the States of Paraná and Mato Grosso do Sul, Brazil. METHODS NHP sera samples (total n = 80, Alouatta spp. n = 07, Callithrix spp. n = 29 and Sapajus spp. n = 44) were screened for the presence of viral genomes using reverse transcription polymerase chain reaction and 10% polyacrylamide gel electrophoresis techniques. RESULTS All of the samples were negative for the Flavivirus genome following the 10% polyacrylamide gel electrophoresis analysis. CONCLUSIONS These negative results indicate that the analyzed animals were not infected with arboviruses from the Flavivirus genus and did not represent a risk for viral transmission through vectors during the period in which the samples were collected.