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1.
Mem. Inst. Oswaldo Cruz ; 114: e180598, 2019. tab, graf
Article in English | LILACS | ID: biblio-1040630

ABSTRACT

Anopheles bellator is a primary malaria vector in the Atlantic Forest. Partial sequences of timeless and Clock genes were used to assess the genetic differentiation of five Brazilian populations, which showed strong population structure (e.g. high F ST values and fixed differences) in all pairwise comparisons between Bahia sample and the others from Paraná, São Paulo and Rio de Janeiro states. Also, the resulting phylogenetic trees clearly grouped the sequences from Bahia in a different cluster with high bootstrap values. Among southern and southeastern populations low levels of genetic differentiation were found suggesting a general stability of the genetic structure.


Subject(s)
Animals , Genetic Variation/genetics , Mosquito Vectors/genetics , Anopheles/genetics , Phylogeny , Brazil , Forests , Sequence Analysis, DNA , Mosquito Vectors/classification , Malaria/transmission , Anopheles/classification
2.
Biomédica (Bogotá) ; 38(supl.2): 117-126, ago. 2018. tab, graf
Article in Spanish | LILACS | ID: biblio-974013

ABSTRACT

Introducción. A pesar de los recientes reportes de infección con Plasmodium spp. en poblaciones relacionadas con los linajes noroeste y sureste, Anopheles triannulatus no está incriminado como vector de la transmisión de malaria en Colombia. La diversidad genética puede delimitar la información sobre el flujo génico y la diferenciación poblacional entre localidades con malaria. Objetivo. Estimar la diversidad genética de An. triannulatus en cinco municipios con alta y baja incidencia de malaria en el departamento de Córdoba. Materiales y métodos. La recolección entomológica se hizo entre agosto y noviembre de 2016 en los municipios de Tierralta, Puerto Libertador, Montelíbano, Sahagún y Planeta Rica. Como marcador genético, se utilizó la región de código de barras de ADN del gen mitocondrial COI. El análisis genético incluyó la estimación de los parámetros de diversidad haplotípica, estructura genética y flujo génico, la prueba D de neutralidad de Tajima, la red de haplotipos y las relaciones filogenéticas. Resultados. Se obtuvieron 148 secuencias parciales de 655 nucleótidos del gen COI, de los cuales se derivaron 44 haplotipos. Los haplotipos H2 y H21 fueron los más frecuentes en las poblaciones. Los valores de la prueba D de Tajima fueron negativos y no significativos (p>0,10). Los estimadores de estructura genética (FST=0,01427) y de flujo génico (Nm=17,27) evidenciaron que no hubo diferenciación genética en las poblaciones muestreadas debido al importante intercambio de migrantes. Mediante las inferencias filogenéticas y la red de haplotipos, se identificó una sola especie sin diferenciación geográfica o de linajes en el rango geográfico estudiado. Conclusión. La diversidad genética calculada para An. triannulatus en este contexto, indicó que las poblaciones están en un intercambio constante.


Introduction: Anopheles triannulatus is not incriminated as a vector of malaria transmission in Colombia despite recent reports of infection with Plasmodium spp. in populations related to the northwestern and southeastern lineages. Genetic diversity can delimit information about gene flow and population differentiation in localities with malaria. Objective: To estimate the genetic diversity of An. triannulatus in five municipalities with high and low incidence of malaria in the department of Córdoba. Materials and methods: The entomological collections were done between August and November, 2016, in Tierralta, Puerto Libertador, Montelíbano, Sahagún, and Planeta Rica. We used the COI barcoding fragment as molecular marker. The genetic analysis included the estimation of genetic parameters such as the diversity haplotype, the genetic structure, the gene flow, the Tajima's D test, the haplotype network, and the phylogenetic relationship. Results: We obtained 148 sequences with a length of 655 nucleotides of the COI gene, from which we derived 44 haplotypes. The H2 and H21 haplotypes were the most frequent in the populations. The values of the Tajima's D test were negative and not significant (p>0.10). The genetic structure index (FST=0.01427) and the gene flow (Nm=17.27) evidenced no differentiation between sampled populations due to the high exchange of migrants. Using phylogenetic inferences and the haplotype network, we identified one single species without geographic differentiation or lineages in the geographic range studied. Conclusions: The genetic diversity calculated for An. triannulatus in this context indicated stable populations in constant exchange.


Subject(s)
Anopheles/genetics , Genetic Variation , Haplotypes , Colombia , Gene Flow
3.
Salud pública Méx ; 60(1): 48-55, Jan.-Feb. 2018. graf
Article in English | LILACS | ID: biblio-903862

ABSTRACT

Abstract: Objective: To analyze the transcription pattern of neuropeptides in the ontogeny of a malaria vector, the mosquito Anopheles albimanus. Materials and methods: The transcription pattern of Crustacean CardioActive peptide (CCAP), corazonin, Ecdysis Triggering Hormone (ETH), allatostatin-A, orcokinin, Insulin Like Peptide 2 (ILP2), Insulin Like Peptide 5 (ILP5) and bursicon was evaluated using qPCR on larvae (1st - 4th instar), pupae and adult mosquitoes. Results: Unlike in other insects, transcripts of CCAP (70.8%), ETH (60.2%) and corazonin (76.5%) were expressed in 4th instar larvae, probably because these three neuropeptides are associated with the beginning of ecdysis. The neuropeptide ILP2 showed higher transcription levels in other stages and orcokinin decreased during the development of the mosquito. Conclusion: The CCAP, corazonin and ETH neuropeptides are potential targets for the design of control strategies aimed at disrupting An. albiamnus larval development.


Resumen: Objetivo: Describir la expresión de neuropéptidos durante la ontogenia del mosquito vector de la malaria Anopheles albimanus. Material y métodos: Se midió la expresión de CCAP, corazonina, ETH, allatostatina, orcokinina, ILP2, ILP5 y bursicon en larvas de primer (2mm), segundo (4mm), tercer (5mm) y cuarto (6mm) estadio, pupas y mosquitos adultos, mediante qPCR. Resultados. A diferencia de otros insectos en donde, CCAP, corazonina y ETH se expresan principalmente en estadios pupales, en An. albimanus se expresaron mayoritariamente en larvas de cuarto estadio, CCAP tuvo 70.8% de expresión relativa, corazonina 76.5% y ETH 60.2%. ILP2 fue el neuropéptido que más se expresó en el primer, segundo y tercer estadio y orcokinina disminuyó durante el desarrollo del mosquito. Conclusión. Los péptidos estudiados se expresaron en todos los estadios de desarrollo del mosquito. Sin embargo, su expresión varió en cada uno de ellos. Los neuropéptidos CCAP, corazonina y ETH, que son esenciales para la transformación de lavas a pupas, pueden ser blancos potenciales para el diseño de estrategias de control dirigidas a interrumpir el desarrollo larvario de An. albimanus.


Subject(s)
Animals , Neuropeptides/biosynthesis , Molting/genetics , Insect Proteins/biosynthesis , Anopheles/genetics , Transcription, Genetic , Neuropeptides/genetics , Gene Expression Regulation, Developmental , Insect Proteins/genetics , Real-Time Polymerase Chain Reaction , Larva , Malaria , Anopheles/growth & development
4.
Salud pública Méx ; 60(1): 77-85, Jan.-Feb. 2018. tab, graf
Article in English | LILACS | ID: biblio-903841

ABSTRACT

Abstract: Objective: To analyze the current knowledge of pathogen-insect interactions amenable for the design of molecular-based control strategies of vector-borne diseases. Materials and methods: We examined malaria, dengue, and Chagas disease pathogens and insect molecules that participate in interactions during their vectors infection. Results: Pathogen molecules that participate in the insect intestine invasion and induced vector immune molecules are presented, and their inclusion in transmission blocking vaccines (TBV) and in genetically modify insect (GMI) vectors or symbiotic bacteria are discussed. Conclusion: Disruption of processes by blocking vector-pathogen interactions provides several candidates for molecular control strategies, but TBV and GMI efficacies are still limited and other secondary effects of GMI (improving transmission of other pathogens, affectation of other organisms) should be discarded.


Resumen: Objetivo: Analizar el conocimiento actual de las interacciones patógeno-insecto susceptibles a incluirse en el diseño de estrategias moleculares para el control de enfermedades transmitidas por vectores. Material y métodos: Se examinaron los agentes causales de la malaria, el dengue y la enfermedad de Chagas, y las moléculas de insectos que participan en interacciones durante la infección de sus vectores. Resultados: Se presentan moléculas de patógenos que participan en la invasión del intestino del insecto y moléculas inmunes inducidas en los vectores. Se discute su inclusión en vacunas bloqueadoras de transmisión (VBT) y en la modificación genética de vectores (MGI) o de sus bacterias simbióticas. Conclusión: La interrupción de procesos mediante el bloqueo de las interacciones patógeno-vector proporciona varios candidatos para las estrategias de control molecular, pero la eficacia de VBT y MGI es aún limitada y los efectos secundarios de MGI (aumento de la transmisión de otros patógenos y afectación de otros organismos) deben descartase.


Subject(s)
Animals , Insect Control/methods , Chagas Disease/prevention & control , Dengue/prevention & control , Dengue Virus/physiology , Host-Pathogen Interactions/genetics , Malaria/prevention & control , Plasmodium/physiology , Trypanosoma cruzi/physiology , Aedes/genetics , Reduviidae/genetics , Reduviidae/virology , Mosquito Vectors/genetics , Anopheles/genetics
5.
Ciênc. saúde coletiva ; 21(7): 2179-2188, Jul. 2016.
Article in English | LILACS | ID: lil-785920

ABSTRACT

Abstract The resistance of some species of Anopheles to chemical insecticides is spreading quickly throughout the world and has hindered the actions of prevention and control of malaria. The main mechanism responsible for resistance in these insects appears to be the target site known as knock-down resistance (kdr), which causes mutations in the sodium channel. Even so, many countries have made significant progress in the prevention of malaria, focusing largely on vector control through long-lasting insecticide nets (LLINs), indoor residual spraying and (IRS) of insecticides. The objective of this review is to contribute with information on the more applied insecticides for the control of the main vectors of malaria, its effects, and the different mechanisms of resistance. Currently it is necessary to look for others alternatives, e.g. biological control and products derived from plants and fungi, by using other organisms as a possible regulator of the populations of malaria vectors in critical outbreaks.


Resumo A resistência de algumas espécies de Anopheles a inseticidas químicos está se espalhando rapidamente por todo o mundo e tem dificultado as ações de prevenção e controle da malária. O principal mecanismo responsável pela resistência nestes insetos parece ser o sítio-alvo conhecido como resistência knock-down resistance (kdr), que causa mutações no canal de sódio. Mesmo assim, muitos países fizeram progressos significativos na prevenção da malária, concentrando-se em grande parte no controle do vetor através redes inseticidas de longa duração (RILD), e de pulverização residual interna (PRI) de inseticidas. O objetivo desta revisão é contribuir com informações sobre os inseticidas mais aplicados para o controle dos principais vetores da malária, seus efeitos, e os diferentes mecanismos de resistências. Atualmente é necessário olhar para outras alternativas, como por exemplo, controle biológico e produtos derivados de plantas e fungos, pela utilização de outros organismos como um possível regulador de populações de vetores da malária em surtos críticos.


Subject(s)
Animals , Anopheles/drug effects , Anopheles/genetics , Insecticide Resistance , Insecticides/pharmacology , Agriculture
6.
Mem. Inst. Oswaldo Cruz ; 111(5): 335-346, May 2016. tab, graf
Article in English | LILACS | ID: lil-782048

ABSTRACT

Salivary gland polytene chromosomes of 4th instar Anopheles darlingi Root were examined from multiple locations in the Brazilian Amazon. Minor modifications were made to existing polytene photomaps. These included changes to the breakpoint positions of several previously described paracentric inversions and descriptions of four new paracentric inversions, two on the right arm of chromosome 3 and two on the left arm of chromosome 3 that were found in multiple locations. A total of 18 inversions on the X (n = 1) chromosome, chromosome 2 (n = 7) and 3 (n = 11) were scored for 83 individuals from Manaus, Macapá and Porto Velho municipalities. The frequency of 2Ra inversion karyotypes in Manaus shows significant deficiency of heterozygotes (p < 0.0009). No significant linkage disequilibrium was found between inversions on chromosome 2 and 3. We hypothesize that at least two sympatric subpopulations exist within the An. darlingi population at Manaus based on inversion frequencies.


Subject(s)
Animals , Anopheles/genetics , Chromosome Inversion/genetics , Insect Vectors/genetics , Polytene Chromosomes/genetics , Salivary Glands , Anopheles/classification , Brazil , Chromosome Mapping , Insect Vectors/classification
7.
Belo Horizonte; s.n; 2015. 136 p.
Thesis in English | ColecionaSUS, LILACS, ColecionaSUS | ID: biblio-942729

ABSTRACT

A malária humana é uma doença provocada por parasitas do gênero Plasmodium, os quais na natureza requerem de um mosquito anofelíno para completar o seu ciclo de vida e serem transmitidos a um hospedeiro humano. Nas Américas, o Brasil tem a maior incidência de malária, sendo responsável por 41% dos casos. Com o aparecimento do sequenciamento de nova geração e das ferramentas bioinformática relacionados, grandes avanços foram alcançados em relação à montagem de genomas e transcriptomas de anofelinos, assim como na exploração de estratégias de paratransgenesis para interromper a transmissão da malária. No entanto, os vetores neotropicais da malária encontram-se longe dos vetores da África e Ásia no que refere a estes conhecimentos. Este estudo é parte de um esforço contínuo para montar o genoma do Anopheles aquasalis, um vetor neotropical da malária humana, que atualmente posiciona-se como um excelente modelo de transmissão da malária no Brasil. Em paralelo ao sequenciamento do genoma, e para maximizar os dados gerados, optamos por focar em duas tarefas pontuais e viáveis: explorar a diversidade e composição do consórcio bacteriano associado ao anofelino; assim como montar e caracterizar o genoma mitocondrial desta espécie.


O sequenciamento metagenômico ̈shotgun ̈ e o programa MG-RAST foram utilizados para fazer um ̈screening ̈ das bactérias associadas à pupas de A .aquasalis criadas em laboratório. O consórcio bacteriano predito é composto por 74 gêneros contendo bactérias marinhas e bioluminescentes. No nível taxonômico de família bacteriana, identificamos 14 OTUs compartilhadas entre anofelinos americanos e africanos. Além disso, foram comparadas cinco comunidades bacterianas associadas a duas espécies de anofelinos: A. aquasalis e Anopheles gambiae. Foi identificada uma associação significativa (NPMANOVA p <0,05) entre a composição da comunidade bacteriana e o ambiente aquático (laboratório ou condições semi-naturais) nas quais cada hospedeiro anofelino foi criado. Atualmente, o entendimento da filogenia do gênero Anopheles é limitado e as informações sobre o tempo de divergência dentro da linhagem de mosquitos é escassa.


Apresentamos a sequencia de 15,393 pb correspondente ao genoma mitocondrial de A. aquasalis. Quando comparado com outros mitogenomas anofelinos relevantes, observou-se alta similaridade na composição dos genomas assim como características estruturais conservadas. Através de análises Bayesianas, reconstruímos as relações filogenéticas e estimamos a data de divergência entre 22 anofelinos e outras espécies de dípteros. Descobrimos que o mais recente ancestral entre as subfamílias Nyssorhynchus e Anopheles +Cellia existiu ~ 83 milhões anos atrás (MYA). Estimou-se que A. aquasalis divergiu do complexo do Anopheles albitarisis faz ~ 28 MYA, e faz ~ 38 MYA do Anopheles darlingi. A distribuição estreita e o peculiar nicho ecológico do A. aquasalis, além de considerar a sua adaptação a ambientes larvários com água salobra fizeram nos perguntar se a sua história evolutiva deixou uma marca na arquitetura do seu genoma, assim como sobre a estrutura da comunidade bacteriana associada a este anofelino.


Subject(s)
Male , Female , Humans , Anopheles/genetics , Genome, Mitochondrial/genetics , Malaria/genetics , Microbiota/genetics , Plasmodium/genetics
8.
Mem. Inst. Oswaldo Cruz ; 109(5): 672-684, 19/08/2014. tab, graf
Article in English | LILACS | ID: lil-720419

ABSTRACT

Anophelines harbour a diverse microbial consortium that may represent an extended gene pool for the host. The proposed effects of the insect microbiota span physiological, metabolic and immune processes. Here we synthesise how current metagenomic tools combined with classical culture-dependent techniques provide new insights in the elucidation of the role of the Anopheles-associated microbiota. Many proposed malaria control strategies have been based upon the immunomodulating effects that the bacterial components of the microbiota appear to exert and their ability to express anti-Plasmodium peptides. The number of identified bacterial taxa has increased in the current “omics” era and the available data are mostly scattered or in “tables” that are difficult to exploit. Published microbiota reports for multiple anopheline species were compiled in an Excel® spreadsheet. We then filtered the microbiota data using a continent-oriented criterion and generated a visual correlation showing the exclusive and shared bacterial genera among four continents. The data suggested the existence of a core group of bacteria associated in a stable manner with their anopheline hosts. However, the lack of data from Neotropical vectors may reduce the possibility of defining the core microbiota and understanding the mosquito-bacteria interactive consortium.


Subject(s)
Animals , Anopheles/genetics , Anopheles/microbiology , Insect Vectors/genetics , Metagenomics , Microbiota , Geography, Medical , Microbiota/genetics , Phylogeny
9.
Mem. Inst. Oswaldo Cruz ; 109(4): 473-479, 03/07/2014. tab, graf
Article in English | LILACS | ID: lil-716299

ABSTRACT

Since 1984, Anopheles (Kerteszia) lepidotus has been considered a mosquito species that is involved in the transmission of malaria in Colombia, after having been incriminated as such with epidemiological evidence from a malaria outbreak in Cunday-Villarrica, Tolima. Subsequent morphological analyses of females captured in the same place and at the time of the outbreak showed that the species responsible for the transmission was not An. lepidotus, but rather Anopheles pholidotus. However, the associated morphological stages and DNA sequences of An. pholidotus from the foci of Cunday-Villarrica had not been analysed. Using samples that were caught recently from the outbreak region, the purpose of this study was to provide updated and additional information by analysing the morphology of female mosquitoes, the genitalia of male mosquitoes and fourth instar larvae of An. pholidotus, which was confirmed with DNA sequences of cytochrome oxidase I and rDNA internal transcribed spacer. A total of 1,596 adult females were collected in addition to 37 larval collections in bromeliads. Furthermore, 141 adult females, which were captured from the same area in the years 1981-1982, were analysed morphologically. Ninety-five DNA sequences were analysed for this study. Morphological and molecular analyses showed that the species present in this region corresponds to An. pholidotus. Given the absence of An. lepidotus, even in recent years, we consider that the species of mosquitoes that was previously incriminated as the malaria vector during the outbreak was indeed An. pholidotus, thus ending the controversy.


Subject(s)
Animals , Female , Male , Anopheles/anatomy & histology , Anopheles/genetics , Genitalia, Male/anatomy & histology , Anopheles/classification , Base Sequence , Colombia , DNA, Mitochondrial/genetics , DNA, Ribosomal Spacer/genetics , Larva/anatomy & histology , Larva/classification , Larva/genetics , Molecular Sequence Data , Phylogeny
10.
Mem. Inst. Oswaldo Cruz ; 108(5): 605-615, ago. 2013. tab, graf
Article in English | LILACS | ID: lil-680764

ABSTRACT

To evaluate whether environmental heterogeneity contributes to the genetic heterogeneity in Anopheles triannulatus, larval habitat characteristics across the Brazilian states of Roraima and Pará and genetic sequences were examined. A comparison with Anopheles goeldii was utilised to determine whether high genetic diversity was unique to An. triannulatus. Student t test and analysis of variance found no differences in habitat characteristics between the species. Analysis of population structure of An. triannulatus and An. goeldii revealed distinct demographic histories in a largely overlapping geographic range. Cytochrome oxidase I sequence parsimony networks found geographic clustering for both species; however nuclear marker networks depicted An. triannulatus with a more complex history of fragmentation, secondary contact and recent divergence. Evidence of Pleistocene expansions suggests both species are more likely to be genetically structured by geographic and ecological barriers than demography. We hypothesise that niche partitioning is a driving force for diversity, particularly in An. triannulatus.


Subject(s)
Animals , Anopheles/genetics , Genetic Variation/genetics , Insect Vectors/genetics , Anopheles/classification , Brazil , Gene-Environment Interaction , Geography , Insect Vectors/classification , Population Density
11.
Mem. Inst. Oswaldo Cruz ; 108(3): 312-320, maio 2013. tab, graf
Article in English | LILACS | ID: lil-676981

ABSTRACT

To assess differentiation and relationships between Anopheles lesteri and Anopheles paraliae we established three and five iso-female lines of An. lesteri from Korea and An. paraliae from Thailand, respectively. These isolines were used to investigate the genetic relationships between the two taxa by crossing experiments and by comparing DNA sequences of ribosomal DNA second internal transcribed spacer (ITS2) and mitochondrial DNA cytochrome c oxidase subunit I (COI) and subunit II (COII). Results of reciprocal and F1-hybrid crosses between An. lesteri and An. paraliae indicated that they were compatible genetically producing viable progenies and complete synaptic salivary gland polytene chromosomes without inversion loops in all chromosome arms. The pairwise genetic distances of ITS2, COI and COII between these morphological species were 0.040, 0.007-0.017 and 0.008-0.011, respectively. The specific species status of An. paraliae in Thailand and/or other parts of the continent are discussed.


Subject(s)
Animals , Female , Anopheles/genetics , Crosses, Genetic , DNA, Mitochondrial/genetics , DNA, Ribosomal Spacer/genetics , Anopheles/classification , Electron Transport Complex IV/genetics , Karyotyping , Republic of Korea , Sequence Analysis, DNA , Thailand
12.
Indian J Exp Biol ; 2013 Apr; 51(4): 273-279
Article in English | IMSEAR | ID: sea-147592

ABSTRACT

Malaria is a complex disease that afflicts human today. Malaria epidemiology is associated with drug resistance in parasite and differential distribution and insecticide resistance in vector. Efforts are being made to eradicate malaria but burden of malaria is still increasing. Vector control is essential for malaria prevention strategies. Knowledge of population genetic structure is pre-requisite for determining prevention strategies, particularly using transgenic mosquitoes. Population genetic study can predict level of gene flow between different populations. Anopheles stephensi Liston is urban vector of malaria in Indo-Pakistan subcontinent. About 12% of malaria cases of malaria in India are contributed by A. stephensi. Studies conducted on population genetics of A. stephensi using various markers in different parts of the world are discussed in this communication.


Subject(s)
Animals , Anopheles/genetics , DNA, Mitochondrial/genetics , DNA, Ribosomal/genetics , Genetic Markers/genetics , Genetics, Population , Geography , Humans , India , Insect Vectors/genetics , Insecticide Resistance/genetics , Malaria/parasitology , Malaria/prevention & control , Microsatellite Repeats/genetics , Pakistan , Random Amplified Polymorphic DNA Technique
13.
São Paulo; s.n; 2012. 121 p.
Thesis in Portuguese | LILACS | ID: lil-643301

ABSTRACT

Introdução - A malária é uma das principais doenças humanas do mundo e afeta principalmente as populações pobres em regiões tropicais e subtropicais onde as condições ambientais são favoráveis tanto para a proliferação dos agentes etiológicos como dos mosquitos vetores. No Brasil, An. darlingi é considerado vetor primário de plasmódios humanos. Devido à importância médica, esse inseto tem sido objeto de campanhas de controle populacional. No entanto, não foi considerada a ocorrência de microevolução em An. darlingi e consequentemente, a possibilidade de populações diferentes apresentarem características biológicas distintas. Objetivo Verificar a presença de variabilidade genética e morfológica em populações de An. darlingi no Brasil. Metodologia - Foram analisados e comparados 26 atributos da morfologia externa de ovos de nove populações, bem como a morfometria geométrica da forma alar de espécimes de 10 localidades diferentes de Anopheles darlingi. Além disso, foram empregadas sequências do gene mitocondrial citocromo oxidase subunidade I (COI) para analisar a estrutura populacional de An. darlingi. Resultados e Conclusão Apesar dos atributos dos ovos apresentarem variação, somente as amostras de Tocantins e Pará foram diferenciadas das demais populações. As variações nas estruturas externas dos ovos são provavelmente adaptativas, com influência de fatores ambientais, como temperatura, umidade e disponibilidade de alimento para as fêmeas. A comparação da morfometria geométrica da asa demonstrou que existe maior similaridade entre as populações da costa (estados do Espírito Santo , do cerrado (estados de Goiás e Tocantins), do norte do rio Amazonas (estados do Amazonas e Amapá), e do sul do rio Amazonas (estados de Mato Grosso e Pará).


Subject(s)
Animals , Anopheles/anatomy & histology , Anopheles/genetics , Insect Vectors/anatomy & histology , Insect Vectors/genetics , Brazil , Electron Transport Complex IV/genetics , Wings, Animal/anatomy & histology , Zygote/ultrastructure
14.
Mem. Inst. Oswaldo Cruz ; 106(8): 1017-1023, Dec. 2011. graf, mapas, tab
Article in English | LILACS | ID: lil-610980

ABSTRACT

To elucidate the Anopheles nuneztovari s.l. taxonomic status at a microgeographic level in four malaria endemic localities from Antioquia and Córdoba, Colombia, fragments of the cytochrome oxidase subunit I (COI) and the white gene were used. The COI analysis showed low genetic differentiation with fixation index (F ST) levels between -0.02-0.137 and Nm values between 3-∞, indicating the presence of high gene flow among An. nuneztovari s.l. populations from the four localities. The COI network showed a single most common haplotype, type 1 (n = 55), present in all localities, as the likely ancestral haplotype. Analysis of the white gene showed that An. nuneztovari s.l. populations from both departments grouped with haplotypes 19 and 20, which are part of lineage 3 reported previously. The results of the present study suggest that An. nuneztovari s.l. is a single taxon in the area of the present study.


Subject(s)
Animals , Female , Anopheles/genetics , Electron Transport Complex IV/genetics , Genetic Variation/genetics , Insect Vectors/genetics , Anopheles/classification , Anopheles/enzymology , Colombia , Haplotypes , Insect Vectors/classification , Insect Vectors/enzymology , Malaria/transmission , Phylogeny
15.
Mem. Inst. Oswaldo Cruz ; 106(6): 705-715, Sept. 2011. ilus, tab
Article in English | SES-SP, LILACS, SES-SP | ID: lil-602054

ABSTRACT

Phylogenetic relationships among species of the Myzorhynchella Section of Anopheles (Nyssorhynchus) were investigated using the nuclear ribosomal DNA second internal transcribed spacer (ITS2), the nuclear whitegene and mitochondrial cytochrome oxidase subunit I (COI) regions. The recently described Anopheles pristinus and resurrected Anopheles guarani were also included in the study. Bayesian phylogenetic analyses found Anopheles parvus to be the most distantly related species within the Section, a finding that is consistent with morphology. An. pristinus and An. guarani were clearly resolved from Anopheles antunesi and Anopheles lutzii, respectively. An. lutzii collected in the same mountain range as the type locality were found within a strongly supported clade, whereas individuals from the southern state of Rio Grande do Sul, tentatively identified as An. lutzii based on adult female external morphology, were distinct from An. lutzii, An. antunesi and from each other, and may therefore represent two new sympatric species. A more detailed examination of An. lutzii sensu latoalong its known geographic range is recommended to resolve these anomalous relationships.


Subject(s)
Animals , Female , Anopheles/genetics , DNA, Ribosomal Spacer/genetics , Electron Transport Complex IV/genetics , Genes, Insect/genetics , Anopheles/classification , Bayes Theorem , Cell Nucleus/genetics , Mitochondria/enzymology , Phylogeny , Species Specificity
16.
São Paulo; s.n; 2011. 184 p.
Thesis in Portuguese | LILACS | ID: lil-643294

ABSTRACT

Os mosquitos são responsáveis pela transmissão de várias doenças ao homem, entre elas malária, dengue e filarioses. O Laboratório de Biologia Molecular de Vetores do Departamento de Epidemiologia da Faculdade de Saúde Pública da Universidade de São Paulo vem desenvolvendo projetos de pesquisa relacionados aos aspectos moleculares e ao controle genético dos mosquitos vetores, principalmente dos gêneros Anopheles e Culex. A identificação das espécies contidas nesses gêneros é dificultada pela falta de características marcantes nas fêmeas adultas, e pelo fato de muitos desses pertencerem a complexos de espécies crípticas. Estes problemas levaram a utilização de abordagens moleculares, com a utilização de marcadores mitocondriais e ribossômicos para auxiliar na taxonomia e a responder questões filogenéticas. Entre os estudos discutidos nessa tese, é mostrado o Anopheles darlingi como espécie monotípica e o An. oswaldoi como um complexo de espécies. Já os estudos com espécies do gênero Culex indicaram a presença de alguns complexos e de espécies em processos de especiação. Sabe-se que a ocorrência de espécies crípticas entre os vetores de patógenos e o desenvolvimento de técnicas que permitam a correta identificação é crucial em programas de controle. O ressurgimento de parasitoses e arboviroses transmitidas por mosquitos devido principalmente a resistência desses aos inseticidas levaram ao surgimento de estratégias alternativas de controle, tais como a utilização de mosquitos geneticamente modificados para supressão ou substituição de populações vetoras. Embora a competência e a viabilidade da modificação genética de mosquitos tenham sido demonstradas em laboratório, ainda não se sabe como os transgenes podem ser inseridos nas populações de mosquitos na natureza. Foram propostas várias abordagens possíveis, como o uso de elementos de transposição. Porém, o êxito de qualquer mecanismo dependerá em parte do custo no fitness dos mosquitos que é imposto pela presença do transgene e, desse modo, estudos de fitness são essenciais para a seleção de linhagens de mosquitos transgênicos que poderão ser utilizadas em programas de controle.


Subject(s)
Animals , Anopheles/genetics , Mosquito Control/methods , Culex/genetics , Culicidae/genetics , Genetic Markers
17.
Article in English | IMSEAR | ID: sea-135578

ABSTRACT

Background & objectives: Under the national antimalaria programme DDT was introduced in early 1950s for vector control and later hexachloro cyclohexane (HCH) followed by malathion and recently synthetic pyrethroids in 1990s to manage the insecticide resistance in Anopheles culicifacies. Subsequent replacement led to development of multiple resistances in An. culicifacies in Surat district in Gujarat State. Indoor residual spray (IRS) was completely withdrawn in southern villages in Surat in 2002. This study was undertaken in these areas to study the persistence of resistance to DDT, malathion and deltamethrin after sequential withdrawal of IRS with these insecticides at different times. Methods: Susceptibility tests on An. culicifacies were conducted using standard WHO methods and kits. Mortality, knockdown time and lethal times were calculated for An. culicifacies exposed to WHO prescribed diagnostic concentrations of different insecticide impregnated papers. Results: Persistence of DDT-resistance was observed even after 30 yr of its withdrawal from IRS. Similarly, persistence of malathion resistance was also observed after 9 yr of its withdrawal from IRS, while reversal of deltamethrin-resistance was observed very fast within 2-3 yr after its withdrawal from IRS in 2002. Interpretation & conclusion: Present data indicate that the quantum of reversion of insecticide resistance in a population is relative and depends on the genetic stability of the respective resistance genes in the mosquitoes. In the present study withdrawal of pyrethroid-IRS resulted in increased susceptibility against pyrethroids alone and was independent of existence of resistance to insecticides of other groups. This study emphasizes that appropriate rotation of different insecticides; including carbamates may prevent or delay the onset of resistance.


Subject(s)
Animals , Anopheles/drug effects , Anopheles/genetics , Anopheles/physiology , DDT/toxicity , Genetics, Population , India , Insecticide Resistance/genetics , Insecticides/toxicity , Malathion/toxicity , Mortality , Mosquito Control/methods , Nitriles/toxicity , Pyrethrins/toxicity , Regression Analysis , Time Factors
18.
Rev. Soc. Bras. Med. Trop ; 43(4): 409-415, jul.-ago. 2010. mapas, tab
Article in English | LILACS | ID: lil-556007

ABSTRACT

INTRODUCTION: Isoenzymatic analyses were performed involving species of the Nyssorhynchus and Anopheles subgenera in order to estimate the intra and interspecies genetic variability. METHODS: Mosquitoes were caught at different localities in the Amazon region. The collection and rearing of mosquitoes in the laboratory followed specific protocols. For the genetic variability analyses, the technique of horizontal electrophoresis on starch and starch-agarose gel with appropriate buffer systems was used. The alloenzyme variation was estimated using the Biosys-1 software. RESULTS: Out of the 13 loci, eight were polymorphic. Anopheles nuneztovari presented the largest number of alleles per locus, while the smallest number was detected in Anopheles marajoara from Macapá. The largest number of polymorphic loci was found for Anopheles marajoara from Maruanum and the smallest for Anopheles benarrochi (Guayará Mirim). Anopheles darlingi (Macapá) presented the greatest heterozygosity (Ho = 0.167 ± 0.071), while the lowest heterozygosity (Ho = 0.045 ± 0.019) was observed in Anopheles intermedius (Pacoval) of the subgenus Anopheles. Wright's F coefficient revealed considerable genetic structuring between the populations of Anopheles darlingi (Fst = 0.110) and between the populations of Anopheles marajoara (Fst = 0.082). CONCLUSIONS: Considering all the species studied, the genetic distance ranged from 0.008 to 1.114. The greatest distance was between Anopheles mattogrossensis and Anopheles oswaldoi, while the smallest was between the Anopheles benarrochi populations.


INTRODUÇÃO: Análises isoenzimáticas foram realizadas envolvendo espécies dos subgêneros Nyssorhynchus e Anopheles para estimar a variabilidade genética intra e interespecífica. MÉTODOS: Os mosquitos foram capturados em diferentes localidades da região Amazônica. A coleta e a criação dos mosquitos em laboratório foram conforme protocolos específicos. Na análise da variabilidade genética empregou-se a técnica de eletroforese horizontal em géis de amido e amido-agarose com sistemas tampões apropriados. A variação alozímica foi estimada pelo Programa Biosys-1. RESULTADOS: Dos 13 loci, oito foram polimórficos. Anopheles nuneztovari apresentou o maior número de alelos por loco, sendo o menor detectado para Anopheles albitarsis (Macapá). O maior número de loci polimórficos foi detectado em Anopheles marajoara (Maruanum) e o menor, em Anopheles benarrochi (Guayará Mirim). Anopheles darlingi (Macapá) apresentou a maior heterozigosidade (Ho = 0,167 ± 0.071) e a menor (Ho = 0,045 ± 0.019) foi observada em Anopheles intermedius (Pacoval) do subgênero Anopheles. O coeficiente F de Wright evidenciou considerável estruturação genética entre populações de Anopheles darlingi (Fst= 0,110) e entre as populações de Anopheles marajoara (Fst= 0,082). CONCLUSÕES: Considerando todas as espécies estudadas, a distância genética variou de 0,008 a 1,144, onde a maior distância foi entre Anopheles mattogrossensis e Anopheles oswaldoi e a menor, entre as populações de Anopheles benarrochi.


Subject(s)
Animals , Female , Anopheles/genetics , Genetic Variation/genetics , Polymorphism, Genetic/genetics , Anopheles/classification , Anopheles/enzymology , Brazil , Electrophoresis , Isoenzymes/genetics
19.
Bol. malariol. salud ambient ; 50(1): 95-107, jul. 2010. ilus, tab
Article in Spanish | LILACS | ID: lil-630430

ABSTRACT

Se reporta por primera vez una lista de especies de anofelinos y culicinos presentes en el territorio indígena del Bajo Caura, estado Bolívar. Entre larvas y adultos se colectaron en total ocho especies de anofelinos pertenecientes a los subgéneros Anopheles Meigen, Lophopodomyia Antunes, Stethomyia Theobald, Nyssorhynchus Blanchard del género Anopheles Meigen y Chagasia bathana Dyar y 10 géneros de culicinos entre los cuales se identificaron siete especies. Se identificaron y caracterizaron los criaderos con base a la hidrología en arroyo, caño, manantial, laguna, pantano y charco. En colectas de estadíos inmaduros la especie más abundante fue Anopheles triannulatus (Neiva & Pinto), mientras que en colectas de adultos con cebos humanos y trampas CDC la especie más abundante fue el vector de malaria An. darlingi Root. Las trampas de luz ultravioleta resultaron ineficientes para capturar anofelinos y culicinos. Se reporta por primera vez para el estado Bolívar los géneros Coquillettidia Dyar y Johnbelkinia Zavortink, así como las especies Aedes (Ochlerotatus) fulvus (Wiedemann) y Ae. (Och.) serratus (Theobald)


This is the first report of anophelines and culicines species in the indigenous territory of the Lower Caura River, Bolívar State. A total of 8 species of anophelines belonging to the subgenus Anopheles Meigen, Lophopodomyia Antunes, Stethomyia Theobald, Nyssorhynchus Blanchard of the genus Anopheles Meigen and Chagasia bathana Dyar and 10 genera of culicines were collected and 7 species identified. Larval habitats were identified and characterized based on the hidrology in stream, small river, spring, lagoon, swamp and pool. Culicines were also collected in artificial and natural containers. The most abundant species in collections of inmature stages was Anopheles triannulatus (Neiva & Pinto), while in adult collections on human landing catches and CDC light traps the most abundant species was the malaria vector An. darling Root. Ultra violet up draft light traps were inefficient to collect anophelines and culicines. It is reported for the first time the presence in Bolívar State of the genus Coquillettidia Dyar and Johnbelkinia Zavortink, and the species Aedes (Ochlerotatus) fulvus (Wiedemann) and Ae. (Och.) serratus (Theobald)


Subject(s)
Humans , Adult , Anopheles/physiology , Anopheles/genetics , Anopheles/immunology , Anopheles/virology , Epidemiology/classification , Epidemiology/history , Epidemiology/trends , Malaria , Public Health
20.
Mem. Inst. Oswaldo Cruz ; 105(3): 278-285, May 2010. ilus
Article in English | LILACS | ID: lil-547297

ABSTRACT

Anopheles (Nyssorhynchus) pristinus Nagaki & Sallum, n. sp. of the Myzorhynchella Section is described based on morphological characters of adult females, males, fourth-instar larvae, pupae and male genitalia. Anopheles (Nyssorhynchus) antunesi Galvão & Amaral is characterized to fix its identity and distinguish it from An. pristinus. The eggs of An. antunesi are described for the first time. Molecular characterization employing sequences of the COI mitochondrial gene and the ITS2 region of ribosomal DNA are provided for each species. An. antunesi and An. pristinus are compared with morphologically similar species of the Myzorhynchella Section. The results of the present study suggest that the new species has been misidentified as both An. antunesi and Anopheles lutzii Cruz. An. antunesi and An. pristinus are sympatric, occurring at high altitudes in Serra da Mantiqueira, Southeastern Brazil.


Subject(s)
Animals , Female , Male , Anopheles , Anopheles/anatomy & histology , Anopheles/classification , Anopheles/genetics , Anopheles/ultrastructure , DNA, Ribosomal Spacer/genetics , Genes, Insect/genetics , Genes, Mitochondrial/genetics , Genitalia, Male/anatomy & histology , Larva , Pupa , Species Specificity
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