Dynamic of Mayaro Virus Transmission in Aedes aegypti, Culex quinquefasciatus Mosquitoes, and a Mice Model.

Mayaro virus (MAYV) is transmitted by Haemagogus spp. mosquitoes and has been circulating in Amazon areas in the North and Central West regions of Brazil since the 1980s, with an increase in human case notifications in the last 10 years. MAYV introduction in urban areas is a public health concern as infections can cause severe symptoms similar to other alphaviruses. Studies with Aedes aegypti have demonstrated the potential vector competence of the species and the detection of MAYV in urban populations of mosquitoes. Considering the two most abundant urban mosquito species in Brazil, we investigated the dynamics of MAYV transmission by Ae. aegypti and Culex quinquefasciatus in a mice model. Mosquito colonies were artificially fed with blood containing MAYV and infection (IR) and dissemination rates (DR) were evaluated. On the 7th day post-infection (dpi), IFNAR BL/6 mice were made available as a blood source to both mosquito species. After the appearance of clinical signs of infection, a second blood feeding was performed with a new group of non-infected mosquitoes. RT-qPCR and plaque assays were carried out with animal and mosquito tissues to determine IR and DR. For Ae. aegypti, we found an IR of 97.5-100% and a DR reached 100% in both 7 and 14 dpi. While IR and DR for Cx. quinquefasciatus was 13.1-14.81% and 60% to 80%, respectively. A total of 18 mice were used (test = 12 and control = 6) for Ae. aegypti and 12 (test = 8 and control = 4) for Cx. quinquefasciatus to evaluate the mosquito-mice transmission rate. All mice that were bitten by infected Ae. aegypti showed clinical signs of infection while all mice exposed to infected Cx. quinquefasciatus mosquitoes remained healthy. Viremia in the mice from Ae. aegypti group ranged from 2.5 × 108 to 5 × 109 PFU/mL. Ae. aegypti from the second blood feeding showed a 50% IR. Our study showed the applicability of an efficient model to complete arbovirus transmission cycle studies and suggests that the Ae. aegypti population evaluated is a competent vector for MAYV, while highlighting the vectorial capacity of Ae. aegypti and the possible introduction into urban areas. The mice model employed here is an important tool for arthropod-vector transmission studies with laboratory and field mosquito populations, as well as with other arboviruses.

Effects of gamma radiation on the vector competence of Aedes aegypti (diptera: Culicidae) to transmit Zika virus.

One of the limitations of the Sterile Insect Technique (SIT), conventionally performed by ionizing radiation, regards separating males from females, which is not 100% effective. Some irradiated females may be released together with males in the field at SIT. The present study aimed to evaluate the influence of ionizing radiation on the ability of Aedes aegypti mosquitoes to transmit the Zika virus after exposing female pupae to a 40 Gy of gamma radiation. The results suggest that the genetic damage induced by exposure of females to this dose level promotes their total sterility, but it does not influence their vector competence. However, our data point out that ionizing radiation may decrease the proportion of infective mosquitoes.

Potential Nosocomial Infections by the Zika and Chikungunya Viruses in Public Health Facilities in the Metropolitan Area of Recife, Brazil.

Since 2015, the Dengue, Zika, and Chikungunya viruses gained notoriety for their impact in public health in many parts of the globe, including Brazil. In Recife, the capital of Pernambuco State, the introduction of ZIKV impacted human population tremendously, owing to the increase in the number of neurological cases, such as the Guillain−Barré and congenital Zika disorders. Later, Recife was considered to be the epicenter for ZIKV epidemics in Brazil. For arboviral diseases, there are some risk factors, such as climate changes, low socioeconomic conditions, and the high densities of vectors populations, that favor the broad and rapid dispersion of these three viruses in the city. Therefore, continuous arbovirus surveillance provides an important tool for detecting these arboviruses and predicting new outbreaks. The purpose of the present study was to evaluate the circulation of DENV, ZIKV, and CHIKV by RT-qPCR in mosquitoes collected in health care units from the metropolitan area of Recife (MAR), during 2018. A total of 2321 female mosquitoes (357 pools) belonging to two species, Aedes aegypti and Culex quinquefasciatus, were collected from 18 different healthcare units, distributed in five cities from the MAR. Twenty-three pools were positive for ZIKV, out of which, seventeen were of C. quinquefasciatus and six were of A. aegypti. Positive pools were collected in 11/18 health care units screened, with Cq values ranging from 30.0 to 37.4 and viral loads varying from 1.88 × 107 to 2.14 × 109 RNA copies/mL. Nosocomial Aedes- and Culex-borne transmission of arbovirus are widely ignored by surveillance and vector control programs, even though healthcare-associated infections (HAI) are considered a serious threat to patient safety worldwide. Although the results presented here concern only the epidemiological scenario from 2018 in MAR, the potential of hospital-acquired transmission through mosquito bites is being overlooked by public health authorities. It is, therefore, of the ultimate importance to establish specific control programs for these locations.

Effects of gamma radiation on the reproductive viability of Aedes aegypti and its descendants (Diptera: Culicidae).

This work evaluated the genetic damage in descendants of male pupae of Aedes (Stegomyia) aegypti (Diptera: Culicidae) separately exposed to 20, 30, and 40 Gy of gamma radiation in the context of Sterile Insect Technique (SIT). Despite the transmission of the dominant lethal mutation, the employed dose levels did not promote a marked reduction in adult mosquito emergence and fertility. This study emphasized that semi-sterilizing doses < 50 Gy for SIT of Aedes aegypti are not recommended.

First report of the L1014F kdr mutation in wild populations of Anopheles arabiensis in Cabo Verde, West Africa.

BACKGROUND: Due to the lack of vaccines, malaria control mainly involves the control of anopheline vectors (Anopheles spp.) using chemical insecticides. However, the prolonged and indiscriminate use of these compounds has led to the emergence of resistance in Anopheles populations in Africa. Insecticide resistance surveillance programs are less frequent in Cabo Verde than in other African countries. This study aimed to investigate the circulation of the L1014F and L1014S alleles in natural populations of Anopheles arabiensis collected from two sampling sites in the city of Praia, Cabo Verde. METHODS: Anopheles larvae were collected from the two sampling sites and reared in the laboratory until the adult stage. Mosquitoes were first morphologically identified by classical taxonomy and then by molecular species identification using molecular markers. All Anopheles arabiensis were subjected to PCR analysis to screen for mutations associated to resistance in the Nav gene. RESULTS: A total of 105 mosquitoes, all belonging to the Anopheles gambiae complex, were identified by classical taxonomy as well as by molecular taxonomy. Molecular identification showed that 100% of the An. gambiae senso lato specimens analyzed corresponded to An. arabiensis. Analysis of the Nav gene revealed the presence of L1014S and L1014F alleles with frequencies of 0.10 and 0.19, respectively. CONCLUSIONS: Our data demonstrated, for the first time, the presence of the L1014F allele in the An. arabiensis population from Cabo Verde, as well as an increase in the frequency of the kdr L1014S allele reported in a previous study. The results of this study demonstrate the need to establish new approaches in vector control programs in Cabo Verde.

Aedes aegypti continuously exposed to Bacillus thuringiensis svar. israelensis does not exhibit changes in life traits but displays increased susceptibility for Zika virus.

BACKGROUND: Aedes aegypti can transmit arboviruses worldwide, and Bacillus thuringiensis svar. israelensis (Bti)-based larvicides represent an effective tool for controlling this species. The safety of Bti and lack of resistance have been widely reported; however, little is known regarding the impact of the extensive use of these larvicides on the life traits of mosquitoes. Therefore, this study investigated biological parameters, including susceptibility to arbovirus, of an Ae. aegypti strain (RecBti) subjected to 29 generations of exposure to Bti compared with the RecL reference strain. METHODS: The biological parameters of individuals reared under controlled conditions were compared. Also, the viral susceptibility of females not exposed to Bti during their larval stage was analysed by oral infection and followed until 14 or 21 days post-infection (dpi). RESULTS: RecBti individuals did not display alterations in the traits that were assessed (fecundity, fertility, pupal weight, developmental time, emergence rate, sex ratio and haematophagic capacity) compared to RecL individuals. Females from both strains were susceptible to dengue serotype 2 (DENV-2) and Zika virus (ZIKV). However, RecBti females showed significantly higher rates of ZIKV infection compared with RecL females at 7 (90% versus 68%, Chi-square: χ2 = 7.27, df = 1, P = 0.006) and 14 dpi (100% versus 87%, Chi-square: χ2 = 7.69, df = 1, P = 0.005) and for dissemination at 7 dpi (83.3% versus 36%, Fisher's exact test: P < 0.0001, OR = 0.11, 95% CI 0.03-0.32). Quantification of DENV-2 and ZIKV viral particles produced statistically similar results for females from both strains. CONCLUSIONS: Prolonged exposure of Ae. aegypti larvae to Bti did not alter most of the evaluated biological parameters, except that RecBti females exhibited a higher vector susceptibility for ZIKV. This finding is related to a background of Bti exposure for several generations but not to a previous exposure of the tested females during the larval stage. This study highlights mosquito responses that could be associated with the chronic exposure to Bti in addition to the primary larvicidal effect elicited by this control agent.

Multiple Introductions Followed by Ongoing Community Spread of SARS-CoV-2 at One of the Largest Metropolitan Areas of Northeast Brazil.

Multiple epicenters of the SARS-CoV-2 pandemic have emerged since the first pneumonia cases in Wuhan, China, such as Italy, USA, and Brazil. Brazil is the third-most affected country worldwide, but genomic sequences of SARS-CoV-2 strains are mostly restricted to states from the Southeast region. Pernambuco state, located in the Northeast region, is the sixth most affected Brazilian state, but very few genomic sequences from the strains circulating in this region are available. We sequenced 101 strains of SARS-CoV-2 from patients presenting Covid-19 symptoms that reside in Pernambuco. Phylogenetic reconstructions revealed that all genomes belong to the B lineage and most of the samples (88%) were classified as lineage B.1.1. We detected multiple viral introductions from abroad (likely from Europe) as well as six local B.1.1 clades composed by Pernambuco only strains. Local clades comprise sequences from the capital city (Recife) and other country-side cities, corroborating the community spread between different municipalities of the state. These findings demonstrate that different from Southeastern Brazilian states where the epidemics were majorly driven by one dominant lineage (B.1.1.28 or B.1.1.33), the early epidemic phase at the Pernambuco state was driven by multiple B.1.1 lineages seeded through both national and international traveling.

Sequencing of ZIKV genomes directly from Ae. aegypti and Cx. quinquefasciatus mosquitoes collected during the 2015-16 epidemics in Recife.

Zika virus (ZIKV) is a negative sense RNA virus from the Flaviviridae family, which was relatively unknown until the first human epidemic in Micronesia, in 2007. Since then, it spread to French Polynesia and the Americas. Recife, the capital of Pernambuco state and epicenter of the Zika epidemic in Brazil, experienced a large number of microcephaly cases and other congenital abnormalities associated to the ZIKV infection from, 2015 to 16. Evidences suggest that both Aedes aegypti and Culex quinquefasciatus mosquitoes from Recife are capable of replicating and transmitting the virus. Here, we conducted high throughput sequencing of ZIKV genomes directly from Ae. aegypti and Cx. quinquefasciatus mosquitoes collected during the ZIKV epidemics in Recife, in order to investigate the variability and evolution of the virus. We obtained 11 draft ZIKV genomes derived from 5 pools from each Ae. aegypti and Cx. quinquefasciatus species. Genome coverage breadth ranged from 16 to 100% and average depth from 45 to 46,584×. Two of these genomes were obtained from pools of Cx. quinquefasciatus females with no sign of blood in the abdomen. Amino acid substitutions found here were not species-specific. In addition, molecular clock dating estimated that ZIKV draft genomes obtained here were co-circulating in other regions of the country during the epidemics. Overall results highlight that viral mutations and even minor variants can be detected in genomes directly sequenced from mosquito samples and insights about natural viral genomic variability and viral evolution can be useful when designing tools for mosquito control programs.

Zika virus detection, isolation and genome sequencing through Culicidae sampling during the epidemic in Vitória, Espírito Santo, Brazil.

BACKGROUND: Zika virus (ZIKV) has been isolated from many mosquito species in nature, but it is believed that the main vectors in urban environments are species of the genus Aedes. Here, we detected and isolated ZIKV in samples from Aedes aegypti, Aedes taeniorhynchus and Culex quinquefasciatus, collected during the Zika epidemic in Vitória, southeast Brazil. Using quantitative real-time polymerase chain reaction, ZIKV detection was performed in mosquito samples collected from February to April 2016. RESULTS: Overall, six pools of mosquitoes were positive for ZIKV: four of Cx. quinquefasciatus, one of Ae. aegypti and one of Ae. taeniorhynchus. Their genomes were sequenced. CONCLUSIONS: These results support and strengthen the hypothesis that other mosquito species can also be involved in ZIKV transmission.

Detection of alleles associated with resistance to chemical insecticide in the malaria vector Anopheles arabiensis in Santiago, Cabo Verde.

BACKGROUND: Mosquitoes of the Anopheles gambiae complex are the main malaria vectors worldwide. Due to the lack of a vaccine to prevent malaria, the principal way to reduce the impact of this disease relies on the use of chemical insecticides to control its vectors. However, the intensive use of such compounds has led to the emergence of insecticide resistance in several Anopheles populations in Africa. This study aimed to investigate the presence of resistance alleles in an Anopheles arabiensis population from the City of Praia, capital of the Archipelago Cabo Verde, one of the countries on the World Health Organization list of countries that are on a path to eliminate local transmission of malaria. METHODS: Larvae from the Anopheles genus were collected using a one-pint dipper in three areas of City of Praia. Larvae were fed and maintained until the emergence of adult mosquitoes, and these were morphologically identified. In addition, molecular identification was performed using IGS markers and all An. arabiensis samples were subjected to PCR to screen for mutations associated to resistance in the Ace-1, Nav and GSTE2 genes. RESULTS: From a total of 440 mosquitoes collected, 52.3% were morphologically identified as An. gambiae sensu lato (s.l.) and 46.7% as Anopheles pretoriensis. The molecular identification showed that 100% of the An. gambiae s.l. were An. arabiensis. The mutations G119S in the Ace-1 gene and L119F in the GSTE2 gene were screened but not found in any sample. However, sequencing analysis for GSTE2 revealed the presence of 37 haplotypes, 16 polymorphic sites and a high genetic diversity (π = 2.67). The L1014S mutation in the Nav (voltage-gated sodium channel gene) was detected at a frequency of 7.3%. CONCLUSION: This is the first study to investigate the circulation of insecticide resistance alleles in An. arabiensis from Cabo Verde. The circulation of the L1014S allele in the population of An. arabiensis in the city of Praia suggests that pyrethroid resistance may arise, be quickly selected, and may affect the process of malaria elimination in Cabo Verde. Molecular monitoring of resistance should continue in order to guide the development of strategies to be used in vector control in the study region.

Development and Validation of Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for Rapid Detection of ZIKV in Mosquito Samples from Brazil.

The rapid spread of Zika virus (ZIKV) represents a global public health problem, especially in areas that harbor several mosquito species responsible for virus transmission, such as Brazil. In these areas, improvement in mosquito control needs to be a top priority, but mosquito viral surveillance occurs inefficiently in ZIKV-endemic countries. Quantitative reverse transcription PCR (qRT-PCR) is the gold standard for molecular diagnostic of ZIKV in both human and mosquito samples. However, the technique presents high cost and limitations for Point-of-care (POC) diagnostics, which hampers its application for a large number of samples in entomological surveillance programs. Here, we developed and validated a one-step reverse transcription LAMP (RT-LAMP) platform for detection of ZIKV in mosquito samples. The RT-LAMP assay was highly specific for ZIKV and up to 10,000 times more sensitive than qRT-PCR. Assay validation was performed using 60 samples from Aedes aegypti and Culex quinquefasciatus mosquitoes collected in Pernambuco State, Brazil, which is at the epicenter of the Zika epidemic. The RT-LAMP had a sensitivity of 100%, specificity of 91.18%, and overall accuracy of 95.24%. Thus, our POC diagnostics is a powerful and inexpensive tool to monitor ZIKV in mosquito populations and will allow developing countries to establish better control strategies for this devastating pathogen.

Sensitivity of RT-PCR method in samples shown to be positive for Zika virus by RT-qPCR in vector competence studies

Abstract Tissue samples from mosquitoes artificially infected with Zika virus and shown to be positive by RT-qPCR were reexamined by RT-PCR. Using these samples we compared the two methods employed in virus RNA detection for vector competence studies. Results demonstrated that, albeit useful, RT-PCR gave false negatives with low viral loads (< 106 RNA copies/ml).

Sensitivity of RT-PCR method in samples shown to be positive for Zika virus by RT-qPCR in vector competence studies.

Tissue samples from mosquitoes artificially infected with Zika virus and shown to be positive by RT-qPCR were reexamined by RT-PCR. Using these samples we compared the two methods employed in virus RNA detection for vector competence studies. Results demonstrated that, albeit useful, RT-PCR gave false negatives with low viral loads (< 106 RNA copies/ml).

Tecnologias integradas para controle biológico, mecânico e genético de Aedes aegypti / Integrated technologies for biological, mechanical and genetic control of Aedes aegypti

Duas tecnologias alternativas para o controle de Aedes aegypti foram avaliadas: a aplicação espacial de larvicida biológico-Bti em potenciais criadouros peridomiciliares, e a liberação de machos estéreis para inviabilização reprodutiva das fêmeas do mosquito. As ações foram realizadas pelos Agentes dos Serviços de Saúde em 15 vilas da Ilha de Fernando de Noronha, e em uma área (900 imóveis) no bairro da Várzea/Recife/Pernambuco. A efetividade dos métodos foi avaliada por indicadores entomológicos,estimados pela presença, quantidade e viabilidade de ovos do mosquito, coletados em armadilhas, e por marcadores genéticos. A aplicação de Bti, com atomizador costal, ocorreu a cada 30 dias em ambas as áreas. Uma redução importante e sustentável da população de A. aegypti,por este método, foi alcançada em 2015/2016 na Várzea e, em 2016, na Ilha, onde a remoção de 18 toneladas de resíduos sólidos em 2015 contribuiu possivelmente para os resultados. Machos esterilizados com radiação gama foram produzidos em massa no laboratório e liberados em uma das vilas da Ilha. A análise espaço-temporal dos indicadores, de dez/2015a ago/2016, revelou redução expressiva da densidade populacional do mosquito e da diversidade genética da população local. Ambas as abordagens parecem ter reduzido o contato homem-vetor e os riscos de transmissão de arboviroses na Ilha, apesar da elevada competência vetorial da população local do mosquito para os vírus Zika e Dengue. Os métodos testados se mostraram eficientes e passíveis de serem integradas às ações do SUS voltadas ao controle de A. aegypti.

Two alternative technologies were evaluated for Aedes aegypti control:the spraying of a biological larvicide (Bti) in potential peridomiciliarybreeding sites and the release of sterile males to promote reproductionblockage in wild females. Actions were carried out by Agents of theHealth Services, in 15 villages of the Fernando de Noronha Island and in 900 properties from the district of Várzea, Recife-PE. The effectiveness of both methods was evaluated by entomological indicators, estimatedby the presence, quantity and viability of eggs from the mosquito collectedin traps and through genetic markers. Bti was delivered by backpacksprayer every 30 days in both areas. A significant and sustainablereduction of the A. aegypti population as a result of this technique wasachieved in 2015/2016 in Várzea and in 2016 in the Island, where it wasstrengthened by the removal of 18 tons of solid waste in 2015. Malessterilized with gamma radiation were mass-produced in the laboratoryand released in one village of the Island. The spatiotemporal analysis ofthe indicators, from Dec/2015 to Aug/2016, revealed a significant reductionin mosquito density, which impacted on the genetic diversity of thelocal population. Both approaches seem to have reduced human-vectorcontact and the risk of arbovirus transmission in the Island, althoughlocal mosquito population presented high vector competence to Zikaand Dengue virus. These methods were efficient and could be integratedinto SUS actions directed to A. aegypti control.

Vector competence of the Aedes aegypti population from Santiago Island, Cape Verde, to different serotypes of dengue virus.

BACKGROUND: Dengue is an arboviral disease caused by dengue virus (DENV), whose main vectors are the mosquitoes Aedes aegypti and Aedes albopictus. A. aegypti is the only DENV vector in Cape Verde, an African country that suffered its first outbreak of dengue in 2009. However, little is known about the variation in the level of vector competence of this mosquito population to the different DENV serotypes. This study aimed to evaluate the vector competence of A. aegypti from the island of Santiago, Cape Verde, to four DENV serotypes and to detect DENV vertical transmission. METHODS: Mosquitoes were fed on blood containing DENV serotypes and were dissected at 7, 14 and 21 days post-infection (dpi) to detect the virus in the midgut, head and salivary glands (SG) using RT-PCR. Additionally, the number of copies of viral RNA present in the SG was determined by qRT-PCR. Furthermore, eggs were collected in the field and adult mosquitoes obtained were analyzed by RT-PCR and the platelia dengue NS1 antigen kit to detect transovarial transmission. RESULTS: High rates of SG infection were observed for DENV-2 and DENV-3 whereas for DENV-1, viral RNA was only detected in the midgut and head. DENV-4 did not spread to the head or SG, maintaining the infection only in the midgut. The number of viral RNA copies in the SG did not vary significantly between DENV-2 and DENV-3 or among the different periods of incubation and the various titers of DENV tested. With respect to DENV surveillance in mosquitoes obtained from the eggs collected in the field, no samples were positive. CONCLUSION: Although no DENV positive samples were collected from the field in 2014, it is important to highlight that the A. aegypti population from Santiago Islands exhibited different degrees of susceptibility to DENV serotypes. This population showed a high vector competence for DENV-2 and DENV-3 strains and a low susceptibility to DENV-1 and DENV-4. Viral RNA copies in the SG remained constant for at least 21 dpi, which may enhance the vector capacity of A. aegypti and suggests the presence of a mechanism modulating virus replication in the SG.

Análise da competência vetorial para o vírus dengue em populações naturais de Aedes aegypti e Aedes albopictus de Pernambuco / Analysis of vector competence for dengue viruses of Aedes aegypti and Aedes albopictus natural populations from Pernambuco

Os mosquitos Aedes aegypti e Aedes albopictus são considerados os principais vetores da dengue, doença que afeta mais de 100 países e atinge, a cada ano, mais de 50 milhões de pessoas. Para que uma espécie seja incriminada como vetor de uma doença, ela deve ser capaz de se infectar com um determinado patógeno via oral, resistir à replicação deste no seu interior e transmiti-lo a um hospedeiro susceptível. Esta habilidade é denominada competência vetorial. Os rápidos avanços da biologia molecular têm facilitado o surgimento de novas abordagens que podem ser utilizadas para avaliar quantitativamente alguns aspectos da competência vetorial. Neste sentido, a RT-PCR quantitativa (qRT-PCR) tem sido amplamente usada em estudos sobre a interação vírus-vetor. O presente estudo visou avaliar o processo de infecção do vírus Dengue (DENV) em amostras de intestinos e glândulas salivares Ae. aegypti e Ae. albopictus, através da quantificação de partículas virais de três sorotipos de DENV via PCR em tempo real. Os resultados mostraram que as três populações de Ae. aegypti (RecLab, Recife e Petrolina) analisadas no presente estudo foram suscetíveis à infecção com DENV-1, DENV-2 e DENV-3. De uma maneira geral, as populações de Ae. aegypti de campo (Recife e Petrolina) foram mais competentes para a infecção do que a colônia de laboratório (RecLab). Considerando a espécie Ae. albopictus, as duas populações (laboratório e campo) também foram suscetíveis à infecção com os três sorotipos, porém a colônia de laboratório foi mais suscetível à infecção do que a população de campo. Os dados obtidos no presente estudo indicam que diferenças na carga viral usada durante a alimentação artificial podem influenciar nas taxas de infecção do vírus no intestino, porém não influenciam a disseminação dos vírus para as glândulas salivares. Outro aspecto importante foi o curto período de incubação extrínseco (PIE) apresentado pelas duas espécies, exceto na população de Ae.aegypti de Petrolina, diferente do que é descrito na literatura. Este resultado tem uma grande importância epidemiológica, pois quanto menor o PIE, maior será o tempo que a espécie vai ser capaz de transmitir o vírus para um hospedeiro sadio, aumentando assim as chances de transmissão de dengue. Além disso, revela a necessidade da implementação de estratégias mais rápidas para bloquear a transmissão da doença. Estas informações serão necessárias para os estudos de avaliação de risco de transmissão da dengue.

Prevalence of the bla (SHV) gene in Klebsiella pneumoniae isolates obtained from hospital and community infections and from the microbiota of healthy individuals in Recife, Brazil.

The aim of this study was to determine the prevalence of the bla (SHV) gene in Klebsiella pneumoniae isolates from hospital and community infections and from the normal microbiota of healthy individuals in Recife, PE, Brazil. Fifty-two K. pneumoniae isolates were analyzed regarding the presence of the bla (SHV) gene, using PCR, and eight isolates were analyzed by DNA sequencing. This gene was detected in 16 isolates from hospital infections, four from community infections, and nine from the normal microbiota. This was the first study to find the bla (SHV) gene in K. pneumoniae isolates from the normal microbiota. Through DNA sequencing of eight K. pneumoniae isolates from hospital and community infections, with a resistance phenotype indicative of extended-spectrum ß-lactamase production, a new SHV variant named SHV-122 was found. We also detected the presence of bla (SHV-1), bla (SHV-11), bla (SHV-28), and bla (SHV-108). The results show that in Recife, Brazil, K. pneumoniae isolates that presented resistance to oxyimino-ß-lactams had high prevalence and diversity of the bla (SHV) gene. We also conclude that there was a high presence of the bla (SHV) gene among isolates from the normal microbiota of healthy individuals.

Epidemiologia molecular do Aedes albopictus (Díptera: Culicidae) / Molecular epidemiology of Aedes albopictus (Diptera: Culicidae)

O objetivo deste estudo foi caracterizar a estrutura genética de populações naturais de Ae. albopictus coletadas na Região Metropolitana do Recife, no Rio de Janeiro e Minas Gerais e avaliar o seu papel como possível vetor na transmissão do Dengue. Para a análise de variabilidade genética foram utilizados dois genes do DNA mitocondrial: citocromo c oxidase II (COII) e nicotinamida adenina dinucleotídeo desidrogenase subunidade 4 (ND4). Nesta análise utilizando a técnica de SSCP foi identificado um total de oito haplótipos para o gene COII em 252 amostras analisadas e onze haplótipos para o gene ND4 em um total de 273 amostras. Os resultados sobre a análise da variância molecular (AMOVA) para ambos os genes sugeriram que essas populações estão sob forte influência da deriva genética, pois foi identificada uma alta diferenciação genética entre essas populações. Os resultados mostraram que as populações provenientes do município de Moreno foram as mais polimórficas para os dois genes, pois identificamos nestas populações a maioria dos haplótipos encontrados. O município de Moreno é circundado por uma área de mata onde podem existir populações de Ae. albopictus com uma alta variabilidade genética. As populações do Rio de Janeiro e Minas Gerais foram consideradas idênticas, pois apresentaram os mesmos haplótipos para o gene ND4. A análise do DNA mitocondrial associado à técnica de SSCP, constitui uma ferramenta eficiente na caracterização da estrutura genética de populações naturais. Este marcador também poderá ser útil na epidemiologia molecular para traçar rotas de dispersão desta espécie e monitorar o impacto das intervenções dos programas de controle de uma maneira rápida e precisa. Com a finalidade de investigar o envolvimento deste culicídeo na transmissão do vírus dengue foram coletados espécimes de Ae. albopictus em áreas randômicas e em residências onde foram relatados casos com suspeita de Dengue. Estas amostras foram testadas por RT-PCR e isolamento viral em culturas de células C6/36. Dos 217 pools de Ae. aegypti e Ae. albopictus testados para a detecção do vírus Dengue na técnica de RTPCR, 10 pools foram positivos. Destes, 9 pools foram de Ae. aegypti e apenas um pool de Ae. albopictus proveniente do município de Moreno contendo 2 fêmeas coletadas na fase adulta.