Insecticide resistance and genetic structure of Aedes aegypti populations from Rio de Janeiro State,Brazil

Vector control largely relies on neurotoxic chemicals, and insecticide resistance (IR) directly threatens their effectiveness. In some cases, specific alleles cause IR, and knowledge of the genetic diversity and gene flow among mosquito populations is crucial to track their arrival, rise, and spread. Here we evaluated Aedes aegypti populations’ susceptibility status, collected in 2016 from six different municipalities of Rio de Janeiro state (RJ), to temephos, pyriproxyfen, malathion, and deltamethrin. We collected eggs of Ae. aegypti in Campos dos Goytacazes (Cgy), Itaperuna (Ipn), Iguaba Grande (Igg), Itaboraí (Ibr), Mangaratiba (Mgr), and Vassouras (Vsr). We followed the World Health Organization (WHO) guidelines and investigated the degree of susceptibility/resistance of mosquitoes to these insecticides. We used the Rockefeller strain as a susceptible positive control. We genotyped the V1016I and F1534C knockdown resistance (kdr) alleles using qPCR TaqMan SNP genotyping assay. Besides, with the use of Ae. aegypti SNP-chip, we performed genomic population analyses by genotyping more than 15,000 biallelic SNPs in mosquitoes from each population. We added previous data from populations from other countries to evaluate the ancestry of RJ populations.All RJ Ae. aegypti populations were susceptible to pyriproxyfen and malathion and highly resistant to deltamethrin. The resistance ratios for temephos was below 3,0 in Cgy, Ibr, and Igg populations, representing the lowest rates since IR monitoring started in this Brazilian region. We found the kdr alleles in high frequencies in all populations, partially justifying the observed resistance to pyrethroid. Population genetics analysis showed that Ae. aegypti revealed potential higher migration among some RJ localities and low genetic structure for most of them. Future population genetic studies, together with IR data in Ae aegypti on a broader scale, can help us predict the gene flow within and among the Brazilian States, allowing us to track the dynamics of arrival and changes in the frequency of IR alleles, and providing critical information to improving vector control program.     

Body size-associated survival and dispersal rates of Aedes aegypti in Rio de Janeiro

The daily survival rate of Aedes aegypti (L) (Diptera: Culicidae) is one of the most important parameters in mathematical models of dengue transmission. In this report, we evaluate the effect of adult body size on the survival and dispersal rates of male and female Ae. aegypti, the primary dengue vector in Brazil. Independent of larval diet (i.e. size), the daily survival rate of females recaptured in the field was greater than that of males (smaller: t = 5.617; d.f. = 15; P < 0.05; larger: t = 4.241; d.f. = 16; P < 0.05). Larger males lived longer than smaller males (t = 2.2893; d.f. = 7; P < 0.05), but no size effect was observed for females (t =- 0.086; d.f. = 19; P= 0.932). The daily survival rate of smaller females was similar to that of larger females (0.712 and 0.730, respectively, as calculated by an exponential model, and 0.743 and 0.779, respectively, calculated by a non-linear model), and they dispersed further than larger females (mean distances travelled were 78.8 m and 40.9 m, respectively; t =- 10.22; d.f. = 323; P < 0.05). Adult body size did not influence male dispersal distances (t = 0.904; d.f. = 206; P= 0.367). Given our evidence that smaller females appear to have similar lifespans and evidence from other studies that they bite more frequently during a single gonotrophic cycle than larger females, our results suggest that smaller females have a higher vectorial capacity.     

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.     

Vectorial competence of Aedes aegypti (Linnaeus 1762) Rio de Janeiro strain, to Dirofilaria immitis (Leidy 1856)

Dirofilaria immitis (Leidy 1856), a nematode parasite, is the etiologic agent of canine heartworm disease and mosquitoes are essential intermediate hosts. Mosquito susceptibility to the worms differ with species, strains and also among individuals of the same strain. To evaluate the degree of susceptibility of Rio de Janeiro laboratory raised strain of Aedes aegypti, we fed mosquitoes on canine blood with different densities of microfilariae (mf). There was no significant difference in the rate of development among the three different densities of mf. Infective larvae were found in the head and proboscis of all mosquitoes provided bloodmeals with different densities of mf after the 11th day post-infection. The infection rate of mosquitoes after ingestion of blood containing 3,000 mf/ml, 5,000 mf/ml and 7,000 mf/ml were 55.3%, 66.7% and 100%, respectively. The vector efficiency indices ranged from 1.6 to 9.3. The finding of L3 stage larvae, high infection rates and vector efficiency indices suggest that Ae. aegypti, Rio de Janeiro laboratory strain, is a potential vector of D. immitis, although of low efficiency.     

Aedes aegypti resistance to temephos during 2001 in several municipalities in the states of Rio de Janeiro, Sergipe, and Alagoas, Brazil

For more than 30 years temephos, an organophosphate insecticide, has been the sole larvicide used in Brazil in the control of Aedes aegypti. Organophosphates were also used for adult control, being replaced by pyrethroids since 1999. In this same year the Brazilian Health Foundation started the coordination of the Ae. aegypti Insecticide Resistance Monitoring Program. In the context of this program, our group was responsible for the detection of temephos resistance in a total of 12 municipalities in the states of Rio de Janeiro (RJ), Alagoas (AL), and Sergipe (SE) during 2001. In each municipality, a pool of mosquitoes collected from different districts was used, with the exception of Rio de Janeiro city, where eight districts have been separately evaluated. Exposure of larvae to the diagnostic dose of temephos revealed resistance in all localities examined, with mortality levels ranging from 4% (Pilares district, Rio de Janeiro, RJ) to 61.9% (Campos dos Goytacazes, RJ). Quantification of mortality showed resistance ratios from 6.1 (Aracaju, SE) to 16.8 (S?o Gon?alo, RJ and Penha district, Rio de Janeiro, RJ). The national dengue control program is presently using these data to subside insecticide resistance management.     

Parity and ovarian development of Aedes aegypti and Ae. albopictus (Diptera: Culicidae) in metropolitan Rio de Janeiro.

Vector blood-feeding frequency, parity, and ovarian development are important factors that can influence pathogen transmission. Parity rates of the dengue vectors Aedes aegypti and Ae. albopictus were determined from females collected from August 2002 to July 2004 in metropolitan Rio de Janeiro. A high frequency of parous Ae. aegypti (92.9%, n = 550) and Ae. albopictus (99.1%, n = 320) females suggested high survivorship of both species. A total of 69% of wild-caught Ae. aegypti females had blood in the midgut compared to 19% of Ae. albopictus. For Ae. aegypti, red-colored midgut contents were associated with ovaries in early stages of development, and brown-colored midguts were associated with ovaries in late stages of maturation. Ovaries of Ae. aegypti females without blood in the midgut were most frequently in stages I and V of Christophers.     

Dispersal of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in an urban endemic dengue area in the State of Rio de Janeiro,Brazil

Experimental releases of female Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus were performed in August and September 1999, in an urban area of Nova Igua u, State of Rio de Janeiro, Brazil, to estimate their flight range in a circular area of 1,600 m where 1,472 ovitraps were set. Releases of 3,055 Ae. aegypti and 2,225 Ae. albopictus females, fed with rubidium (Rb)-marked blood and surgically prevented from subsequent blood-feeding, were separated by 11 days. Rb was detected in ovitrap-collected eggs by atomic emission spectrophotometry. Rb-marked eggs of both species were detected up to 800 m from the release point. Eggs of Ae. albopictus were more numerous and more heterogeneously distributed in the area than those of Ae. aegypti. Eggs positively marked for Rb were found at all borders of the study area, suggesting that egg laying also occurred beyond these limits. Results from this study suggest that females can fly at least 800 m in 6 days and, if infected, potentially spread virus rapidly.     

Natural skip oviposition of the mosquito Aedes aegypti indicated by codominant genetic markers

This study examines the use of codominant restriction fragment length polymorphism (RFLP) markers to estimate the number of sibling families found within and among oviposition sites used by the mosquito Aedes aegypti (L) (Diptera: Culicidae). Estimates were made using pairwise relatedness (rxy) calculations based on alleles shared between individuals. Genotypes for eight laboratory mosquito families were determined at six RFLP loci and the observed allele frequencies were used to generate simulated distributions of rxy from full-sibling and unrelated pairs of individuals. The midpoint (mp) between the means of the pairwise rxy distributions was used to discriminate full-sibling families from unrelated families. Clusters of individuals with rxy values higher than the mp value were grouped as putative sibling families. This method was tested by calculating actual rxy for all pairwise comparisons of the known laboratory full-sibling and paternal half-sibling families, followed by upgma cluster analysis to group sibling families. The technique was then used for sibling estimations on wild caught mosquitoes collected at three locations in Trinidad, West Indies. From field populations, 35 families were estimated among 122 individuals tested with an average of 6.2 families per container. Members of 19 predicted families clustered as groups across multiple containers, providing molecular evidence for skip-oviposition behaviour in Ae. aegypti females, whereby individual females oviposit in more than one container.     

Viral hepatitis markers in antepartum and postpartum women in Rio de Janeiro,Brazil

A seroprevalence study was carried out among a group of women in Rio de Janeiro to determine the prevalence of different markers for viral hepatitis given the limited data among healthy populations. Blood samples collected and tested from 874 women before or after delivery in a public county maternity hospital demonstrated age to be directly related to markers for hepatitis A virus and hepatitis B virus (HBV) infection. The prevalence of HBV and hepatitis C virus infection were lower than that observed in the blood donor population and might be explained by the younger age group and gender.     

Geographic genetic variation in populations of the dengue virus vector Aedes aegypti

Isoenzyme variation was assessed in 79 mosquito samples of Aedes aegypti, and susceptibility to a dengue 2 virus strain was evaluated in 83 samples. Analysis of FST values, differentiation indexes, and geographic distances separating populations revealed that genetic differences between populations depended on the species' history of migration and colonization. Three major clusters were identified: (1). the sylvan form, Ae. ae. formosus, from West Africa and some islands in the Indian Ocean; (2). the domestic form, Ae. ae. aegypti, from Southeast Asia and South America; and (3). Ae. ae. aegypti populations from the South Pacific islands. Two groups were identified on the basis of susceptibility to the dengue virus: (1). populations with high infection rates, mostly the Ae. ae. aegypti form, and (2). mosquitoes with lower infection rates, specifically Ae. ae. formosus. Other evolutionary and epidemiological implications of the genetic variability of Ae. aegypti are also discussed.     

The genetic diversity and phenotypic characterisation of Streptococcus agalactiae isolates from Rio de Janeiro, Brazil

Streptococcus agalactiae isolates are more common among pregnant women, neonates and nonpregnant adults with underlying diseases compared to other demographic groups. In this study, we evaluate the genetic and phenotypic diversity in S. agalactiae strains from Rio de Janeiro (RJ) that were isolated from asymptomatic carriers. We analysed these S. agalactiae strains using pulsed-field gel electrophoresis (PFGE), serotyping and antimicrobial susceptibility testing, as well as by determining the macrolide resistance phenotype, and detecting the presence of the ermA/B, mefA/E and lnuB genes. The serotypes Ia, II, III and V were the most prevalent serotypes observed. The 60 strains analysed were susceptible to penicillin, vancomycin and levofloxacin. Resistance to clindamycin, chloramphenicol, erythromycin, rifampin and tetracycline was observed. Among the erythromycin and/or clindamycin resistant strains, the ermA, ermB and mefA/E genes were detected and the constitutive macrolides, lincosamides and streptogramin B-type resistance was the most prevalent phenotype observed. The lnuB gene was not detected in any of the strains studied. We found 56 PFGE electrophoretic profiles and only 22 of them were allocated in polymorphism patterns. This work presents data on the genetic diversity and prevalent capsular serotypes among RJ isolates. Approximately 85% of these strains came from pregnant women; therefore, these data may be helpful in developing future prophylaxis and treatment strategies for neonatal syndromes in RJ.     

Variation in Aedes aegypti (Diptera: Culicidae) container productivity in a slum and a suburban district of Rio de Janeiro during dry and wet seasons

Seasonal variation in container productivity and infestation levels by Aedes aegypti were evaluated in two areas with distinct levels of urbanization degrees in Rio de Janeiro, a slum and a suburban neighborhood. The four most productive containers can generate up to 90% of total pupae. Large and open-mouthed containers, such as water tanks and metal drums, located outdoors were the most productive in both areas, with up to 47.49% of total Ae. aegypti pupae collected in the shaded sites in the suburban area. Water-tanks were identified as key containers in both areas during both the dry and rainy seasons. Container productivity varied according to seasons and urbanization degree. However, the mean number of pupae per house was higher in the suburban area, but not varied between seasons within each area (P > 0.05). High infestation indexes were observed for both localities, with a house index of 20.5-21.14 in the suburban and of 9.56-11.22 in the urban area. This report gives potential support to a more focused and cost-effective Ae. aegypti control in Rio de Janeiro.     

Diphtheria-neutralizing antibody levels in healthy adults from Rio de Janeiro, Brazil

In Brazil, until 2004, the immunization policy against diphtheria involved childhood vaccination with no official routine booster dose administered after 15 years of age. This study assessed functional antibody levels against diphtheria among blood donors. A total of 140 blood samples were collected, and diphtheria antitoxin levels were evaluated by Vero cell neutralization test. The mean age of the population was 34 years old (range: 18-61 years); 37.8% females and 62.2% males. Overall, 30.7% (95%, CI: 23.4-38.7) individuals presented neutralizing antitoxin antibody titers < 0.01 IU/ml; 42.1% (95%, CI: 34.1-50.4) showed values between 0.01-0.09 IU/ml and, 27.1% (95%, CI: 20.2-34.9) had (3) 0.1 IU/ml. In the subgroup of individuals with history of diphtheria immunization during childhood (85%), a number of 28.5% showed unprotective levels of circulating neutralizing antibody (< 0.01 IU/ml). Despite the continuous progress of immunization programs directed to Brazilian population, currently healthy adults remain susceptible to diphtheria.     

Temporal genetic variation in Aedes aegypti populations in Ho Chi Minh City (Vietnam)

Aedes aegypti, the main vector of dengue viruses in Asia, displays variation in population density over time. The larval habitats of this species being unevenly distributed and transient (depending on cycles of drought and flood), the forces generating temporal variation in gene frequencies in populations are studied. We sampled seven mosquito populations from Ho Chi Minh City (Vietnam) and its suburbs on five occasions between April 1999 and August 2000. We investigated genetic variation by studying isoenzyme and microsatellite polymorphism and susceptibility to a dengue 2 virus strain. Ae. aegypti populations collected during the dry season (January-April) showed genetic differentiation (F(ST) = 0.016, P < 10(-6) for isoenzymes) and showed more differentiated infection rates of the dengue 2 virus. The genetic structure of the population is less marked during the rainy season (F(ST) = 0.081, P < 10(-6)). Thus, environmental factors, such as rainfall and factors related to human activity, such as breeding site density and insecticide treatment, control the genetic structure of Ae. aegypti populations in the short term. The implications of studies of this kind for the design of future control programmes are discussed.     

The invasion of urban forest by dengue vectors in Rio de Janeiro.

The invasion of a secondary forest within the city of Rio de Janeiro by Aedes aegypti and Ae. albopictus was evaluated from July 1997 to June 1998 through collections of immature stages in ovitraps set at 1 m, 10 m, 100 m, 500 m, and 1,000 m into the forest from houses on the periphery. Both mosquito species were much more abundant close to houses (1-10 m). Aedes aegypti was not collected beyond 100 m, while Ae. albopictus was the most abundant species overall and in ovitraps at all distances from houses. Abundances of Ae. albopictus were significantly correlated with time-lagged rainfall and with abundances of Ae. aegypti. Co-occurrences of Ae. albopictus in traps with Ae. aegypti and Limatus durhami, but not with Culex dolosus, were more likely close to houses. The results suggest that the urban forest is a refuge for both Aedes species, but especially for Ae. albopictus, whose abundance both near houses and in the forest raises concern that this invader may transmit arboviruses to humans that are presently restricted to the sylvan environment.     

Loss of genetic diversity in farmed populations of Colossoma macropomum estimated by microsatellites

The genetic variability of four Colossoma macropomum broodstocks, three from fish farms in different regions and one from the natural environment in Brazil, was analyzed using microsatellite markers. The wild progeny (n = 30) were caught in the Solimões–Amazonas River, at the varzea lakes; this location is used to mature the fish from larvae to juveniles. The three fish farms were selected according to the age of their lineages and broodstock availability: DNOCS (n = 21) is located in the Ceará State, representing the oldest lineage of cultivated tambaqui in Brazil; Balbina (n = 30) is located in the Amazonas State, representing the youngest stocks of tambaqui farmed in Brazil (approximately 15 years); and UFRPE (n = 30) is located in the Pernambuco State and is considered to be a mixed stock formed from the DNOCS and Balbina lineages. The analysis of 13 microsatellite loci indicated the occurrence of a variability reduction in the farmed populations; the UFRPE stock was the population with the highest diversity level. Low values of molecular coancestry were found in these populations. Additionally, significant differences in the R_ST values among the populations were detected, as was the occurrence of genetic structure. The genetic loss found in these populations may have been influenced by the founder effect. Because no breeding programs were during the entire production period and no pedigree records were kept for these broodstocks, we suggest that a wild population might be used as an important genetic resource to increase the genetic diversity of renewal stock lineages.     

Bone sporotrichosis: 41 cases from a reference hospital in Rio de Janeiro,Brazil

BackgroundBone sporotrichosis is rare. The metropolitan region of Rio de Janeiro is hyperendemic for zoonotic sporotrichosis and the bone presentations are increasing.MethodsWe studied a retrospective cohort of 41 cases of bone sporotrichosis, diagnosed from 1999–2016. The inclusion criteria was fungal culture isolation from any clinical specimen associated to bone involvement (radiography and/or computed tomography) compatible with fungal osteomyelitis or histopathological findings of bone material compatible with sporotrichosis. Molecular identification was performed when possible.ResultsMale patients represented 58.5% of the cases, with a cohort median age of 43 years. Immunosuppressive conditions were present in 68.3% of the patients, mostly HIV coinfection (51.2%). Multifocal bone involvement (more than one anatomical segment) was diagnosed in 61% of the patients, while 39% presented unifocal involvement. The bones of the hands were the most affected (58.5%), followed by the feet (41.5%) and tibia (26.8%). Multifocal group was characterized by a higher proportion of males (p = 0.0045) with immunosuppressive conditions (p = 0.0014). Amphotericin B followed by oral itraconazole was the main treatment, with a median time of 16.7 months (1.5 to 99.2 months), and cure of 53.7% of the patients (84.6% of immunocompetent and 39.3% of immunocompromised patients). Sequelae occurred in 12.2% of the patients—amputations (7.3%) and ankylosis (4.9%), while 22% died in the course of the disease. Sporothrix brasiliensis was the causative agent in all the 9 (22%) performed cases.ConclusionsBone sporotrichosis is a chronic, challenging condition with prolonged treatment, often with poor results and sequelae.