Bacterial microbiomes from vertically-transmitted fungal inocula of the leaf-cutting ant Atta texana

Microbiome surveys provide clues for the functional roles of symbiotic microbial communities and their hosts. In this study, we elucidated bacterial microbiomes associated with the vertically transmitted fungal inocula (pellets) used by foundress queens of the leaf-cutting ant Atta texana as starter-cultures for new gardens. As reference microbiomes, we also surveyed bacterial microbiomes of foundress queens, gardens and brood of incipient nests. Pseudomonas, Acinetobacter, Propionibacterium and Corynebacterium were consistently present in high abundance in microbiomes. Some pellet and ant samples contained abundant bacteria from an Entomoplasmatales-clade, and a separate PCR-based survey of Entomoplasmatales bacteria in eight attine ant-genera from Brazil placed these bacteria in a monophyletic clade within the bacterial genus Mesoplasma. The attine ant-Mesoplasma association parallels a similar association between a closely related, monophyletic Entomoplasmatales-clade and army ants. Of thirteen A. texana nests surveyed, three nests with exceptionally high Mesoplasma abundance died, whereas the other nests survived. It is unclear whether Mesoplasma was the primary cause of mortality, or Mesoplasma became abundant in moribund nests for non-pathogenic reasons. However, the consistent and geographically widespread presence of Mesoplasma suggests an important functional role in the association with attine ants

Perfil de susceptibilidad a Temefos en poblaciones de Aedes aegypti (Diptera: Culicidae) de Ciudad del Este - Alto Paraná, Paraguay

En Paraguay, el control del mosquito Aedes aegypti involucrado en la transmisión de varias arbovirosis implica la utilización del Temefos, un organofosforado que ha sido utilizado por los programas nacionales para el control vectorial por más de dos décadas en busca de la reducción de los estadios larvarios. En vista de la necesidad de evaluar periódicamente la actividad larvicida del compuesto químico mencionado, este estudio tuvo como objetivo monitorear el perfil de susceptibilidad de larvas de Ae. aegypti al Temefos. Para ello se aplicó un estudio analítico experimental con ensayos biológicos tipo dosis - respuesta, utilizando larvas del tercer estadio de la primera generación procedentes de una colonia de mosquitos colectada en Ciudad del Este. Las larvas fueron expuestas a la acción del Temefos a diferentes concentraciones definidas por un pre- test. Se registraron valores correspondientes al número de larvas expuestas y mortalidad al término de cada ensayo. Los resultados fueron concentración letal CL50 = 0,00966 mg/L y CL90 = 0,03015mg/L, a partir de estos valores se obtuvieron los indicadores cuantitativos de resistencia, Razón de resistencia RR50 = 2,3734 y RR90 = 4,1643 respectivamente. Este último es un indicativo de resistencia baja en las poblaciones de Ae. aegypti evaluadas, acorde con rangos estandarizados (RR>3<5). Los resultados observados en las poblaciones silvestres de larvas revelan una situación de alerta, considerando que el presente estudio evidenció un proceso de resistencia incipiente al Temefos. Finalmente, basados en los resultados se recomienda plantear y ejecutar estrategias basadas en acciones que permitan preservar la actividad larvicida de este compuesto, evitando el aumento progresivo de resistencia en las poblaciones silvestres

Detecção de genes diferencialmente expressos na formiga urbana Camponotus vittatus (Hymenoptera, Formicinae)

Social insects inspire great curiosity, interest and a wish to know the social life molecular basis. Differentially expressed genes identification has been used with the objective to understand either the gene function and the molecular mechanisms of different biological processes. In this study, two protocols for the total RNA extraction from Camponotus vittatus were tested and the Differential Display Reverse Transcriptase Polymerase Chain Reaction (DDRT-PCR) technique was used with the aim to identify differentially expressed genes in caste, sex and development stages of this ant. The RNA extraction method, based in Chomczynski e Sacchi (1987), was the most efficacious. The DDRT-PCR technique was efficient for the detection of differentially expressed genes from C. vittatus. Three expressed sequence tags (ESTs) were sequenced and the analysis with bioinformatics tools failed in detect similarity with those sequences from data banks. These sequences may be new and, perhaps, specific of C. vittatus. They were deposited in dbEST database. A differentially expressed fragment, specific of works, was obtained with the primers HT11G and AP03. The results of its sequence and the analysis by bioinformatics revealed that this EST was, in fact, been isolated from the endosymbiotic bacteria of C. vittatus because a grol domain was detected and was observed high similarity with many proteins of the chaperonin family, a kind of protein present in the endosymbiotic bacterias of this genus.Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorMestre em Genética e BioquímicaInsetos sociais despertam grande curiosidade, interesse e uma vontade de se compreender as bases moleculares da vida social. A identificação de genes diferencialmente expressos tem sido usada para o entendimento não só da função gênica, mas também dos mecanismos moleculares de diferentes processos biológicos. Nesse estudo, foram testados dois protocolos para extração de RNA total de Camponotus vittatus e a técnica de Differential Display Reverse Transcriptase Polymerase Chain Reaction (DDRT-PCR) foi utilizada para a identificação de genes diferencialmente expressos em casta, sexo e nos estágios de desenvolvimento dessa formiga. O método de extração de RNA baseado em Chomczynski e Sacchi (1987) demonstrou ser o mais eficaz. A técnica DDRTPCR foi eficiente para a detecção de genes diferencialmente expressos de C. vittatus. Três expressed sequence tags (ESTs) foram seqüenciadas e análises por bioinformática não detectaram similaridade com seqüências depositadas em banco de dados, podendo ser novas e, talvez, específicas de C. vittatus. Essas seqüências foram depositadas no banco de dados dbEST. Um fragmento diferencialmente expresso em operária foi obtido com a combinação dos primers: HT11G e AP03. Os resultados do seqüenciamento e das análises por bioinformática revelaram que tal EST foi isolada da bactéria endossimbionte de C. vittatus, pois foi detectada a presença de um domínio grol e alta similaridade com diversas proteínas da família das chaperoninas, proteínas presentes nas bactérias endossimbiontes do gênero

Formigas urbanas como vetores de propagação bacteriana no Hospital de Clínicas da Universidade Federal de Uberlândia

Trabalho de Conclusão de Curso (Graduação)Este trabalho teve como objetivo verificar a presença de bactéricas em formigas urbanas colhidas em setores do HC-UFU, no Campus e em residências, bem como avaliar o perfil de resistência dessas bactérias frente a alguns antimicrobianos analisando, inclusive, amostras retiradas dos ambientes de coleta a partir de esfregaço com swab

Caracterização do transcriptoma e genoma mitocondrial da formiga cortadeira Atta laevigata (Formicidae : Attini)

Formigas cortadeiras do gênero Atta, popularmente conhecidas como saúvas, são as mais derivadas dentro da tribo Attini. Apresentam grande importância ecológica, porém, pelo hábito de cortarem folhas para manutenção do fungo simbionte e pelo enorme tamanho das colônias, causam muitos prejuízos às lavouras, pastagens e plantações, sendo consideradas pragas agrícolas. Atta laevigata Smith, 1858 apresenta vasta distribuição pelo Brasil e é responsável pela herbivoria de inúmeras plantas dicotiledôneas, gramíneas e espécies nativas de diferentes biomas. O presente trabalho teve como objetivos a caracterização parcial do transcriptoma e do genoma mitocondrial de A. laevigata. Foram caracterizadas 2006 sequências únicas do transcriptoma, a partir de uma biblioteca de cDNA preparada com indivíduos inteiros da formiga. Entre essas sequências, 16 provavelmente representam genes com grande número de transcritos. Esses 16 genes estão relacionados a três funções celulares: (i) conservação de energia através de reações redox na mitocôndria; (ii) estrutural, pelo citoesqueleto e músculos; (iii) regulação da expressão gênica e metabolismo. Considerando o estilo de vida e processos biológicos chaves para essas formigas, 146 sequências foram identificadas com base na sua utilização para o controle de cortadeiras pragas. A partir de dados da biblioteca de cDNA e procedimentos envolvendo primer walking, o genoma mitocondrial de A. laevigata foi parcialmente caracterizado, apresentandose com 17920 pb, maior, portanto, do que outros já descritos em Hymenoptera, mesmo considerando-se a impossibilidade de determinação da sequência de uma pequena porção do mtDNA, envolvendo a região controle, uma parte do 12S e os tRNAs S1, V e M. Como já descrito para outros mitogenomas, o de A. laevigata apresentou alto conteúdo AT, os mesmos 13 genes codificadores...Leafcutter ants from Atta genus, popularly known as “saúvas”, are the most derived of the tribe Attini. They have major ecological importance, but, because of their habit of cutting leaves for the maintenance of the symbiotic fungus and the huge colony size, they impose severe economic damages to plantations, pastures, and agriculture, being considered as agriculture pests. Atta laevigata shows wide distribution in Brazil and it is responsible for the herbivory of many dicots, grass, and native species from different biomes. The present work aimed to characterize the transcriptome and the mitochondrial genome of A. laevigata. 2,006 unique sequences of the transcriptome were characterized from a cDNA library constructed with whole individuals. Among those sequences, 16 are likely from genes with high number of transcripts. Those 16 genes are related with three cellular functions: (i) energy conservation through redox reactions in mitochondria; (ii) cytoskeleton and muscle structuring; (iii) regulation of gene expression and metabolism. Based on lifestyle and key biological processes of these ants, 146 sequences were identified with potential use for controlling pest leafcutters. Using data from cDNA library and primer walking proceedings, the mitochondrial genome of A. laevigata was partially characterized with 17,920 bp, being larger than the others already described for Hymenoptera. A small part of the mtDNA was not sequenced, including the control region, a portion of 12S and tRNAs S1, V, and M. As described before for other mitogenomes, A. laevigata mtDNA displayed high AT contain, the same 13 proteincoding genes and the two ribosomal subunits with length and location according to the hypothetic ancestral mitogenome. Rearrangements were found for the tRNAs, but the most remarkable difference were the high number and longer length of intergenic regions presented in the mtDNA... (Complete abstract click electronic access below)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

The Mitochondrial Genome of the Leaf-Cutter Ant Atta laevigata: A Mitogenome with a Large Number of Intergenic Spacers

In this paper we describe the nearly complete mitochondrial genome of the leaf-cutter ant Atta laevigata, assembled using transcriptomic libraries from Sanger and Illumina next generation sequencing (NGS), and PCR products. This mitogenome was found to be very large (18,729 bp), given the presence of 30 non-coding intergenic spacers (IGS) spanning 3,808 bp. A portion of the putative control region remained unsequenced. The gene content and organization correspond to that inferred for the ancestral pancrustacea, except for two tRNA gene rearrangements that have been described previously in other ants. The IGS were highly variable in length and dispersed through the mitogenome. This pattern was also found for the other hymenopterans in particular for the monophyletic Apocrita. These spacers with unknown function may be valuable for characterizing genome evolution and distinguishing closely related species and individuals. NGS provided better coverage than Sanger sequencing, especially for tRNA and ribosomal subunit genes, thus facilitating efforts to fill in sequence gaps. The results obtained showed that data from transcriptomic libraries contain valuable information for assembling mitogenomes. The present data also provide a source of molecular markers that will be very important for improving our understanding of genomic evolutionary processes and phylogenetic relationships among hymenopterans

The Mitochondrial Genome of the Leaf-Cutter Ant <i>Atta laevigata</i>: A Mitogenome with a Large Number of Intergenic Spacers

<div><p>In this paper we describe the nearly complete mitochondrial genome of the leaf-cutter ant <i>Atta laevigata</i>, assembled using transcriptomic libraries from Sanger and Illumina next generation sequencing (NGS), and PCR products. This mitogenome was found to be very large (18,729 bp), given the presence of 30 non-coding intergenic spacers (IGS) spanning 3,808 bp. A portion of the putative control region remained unsequenced. The gene content and organization correspond to that inferred for the ancestral pancrustacea, except for two tRNA gene rearrangements that have been described previously in other ants. The IGS were highly variable in length and dispersed through the mitogenome. This pattern was also found for the other hymenopterans in particular for the monophyletic Apocrita. These spacers with unknown function may be valuable for characterizing genome evolution and distinguishing closely related species and individuals. NGS provided better coverage than Sanger sequencing, especially for tRNA and ribosomal subunit genes, thus facilitating efforts to fill in sequence gaps. The results obtained showed that data from transcriptomic libraries contain valuable information for assembling mitogenomes. The present data also provide a source of molecular markers that will be very important for improving our understanding of genomic evolutionary processes and phylogenetic relationships among hymenopterans.</p></div

Is there an efficient trap or collection method for sampling Anopheles darlingi and other malaria vectors that can describe the essential parameters affecting transmission dynamics as effectively as human landing catches? - A Review

Distribution, abundance, feeding behaviour, host preference, parity status and human-biting and infection rates are among the medical entomological parameters evaluated when determining the vector capacity of mosquito species. To evaluate these parameters, mosquitoes must be collected using an appropriate method. Malaria is primarily transmitted by anthropophilic and synanthropic anophelines. Thus, collection methods must result in the identification of the anthropophilic species and efficiently evaluate the parameters involved in malaria transmission dynamics. Consequently, human landing catches would be the most appropriate method if not for their inherent risk. The choice of alternative anopheline collection methods, such as traps, must consider their effectiveness in reproducing the efficiency of human attraction. Collection methods lure mosquitoes by using a mixture of olfactory, visual and thermal cues. Here, we reviewed, classified and compared the efficiency of anopheline collection methods, with an emphasis on Neotropical anthropophilic species, especially Anopheles darlingi, in distinct malaria epidemiological conditions in Brazil

Organization of the <i>A. laevigata</i> mitogenome compared with those of the ancestor and other ants.

<p>All protein and rRNA-coding genes are in the same direction and position found in other Hymenoptera and hypothetical pancrustacean ancestral sequences. Genes encoded by the N strand are underlined; the remaining genes are encoded by the J strand. The control region of <i>A. laevigata</i> (gray) is incomplete. Shaded genes in pancrustacean ancestral sequence indicate rearrangements and arrows indicate position shifts of tRNA genes compared to it. Black arrow: <i>trnV</i> translocation from the <i>lrRNA-srRNA</i> junction to the <i>srRNA-ND2</i> junction; grey arrow: <i>trnI-trnQ-trnM</i> became <i>trnM-trnI-trnQ</i>; blue arrow: <i>trnK</i> and <i>trnD</i> swapped positions; red arrow: <i>trnN</i> translocation from the <i>trnA-trnR-trnN-trnS₁-trnE-trnF</i> cluster to a position upstream of <i>srRNA</i>, with an inversion. This figure was adapted from Gotzek et al. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0097117#pone.0097117-Gotzek1" target="_blank">[13]</a>.</p

Is there an efficient trap or collection method for sampling Anopheles darlingi and other malaria vectors that can describe the essential parameters affecting transmission dynamics as effectively as human landing catches? - A Review

Made available in DSpace on 2015-05-19T13:26:05Z (GMT). No. of bitstreams: 2 license.txt: 1914 bytes, checksum: 7d48279ffeed55da8dfe2f8e81f3b81f (MD5) jose_limaetal_IOC_2014.pdf: 912695 bytes, checksum: 7f717b4969a07d7a9cff1ef6ad248b1a (MD5) Previous issue date: 2014Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Fisiologia e Controle de Artrópodes Vetores. Rio de Janeiro, RJ, Brasil / Instituto de Biologia do Exército. Laboratório de Entomologia. Rio de Janeiro, RJ, Brasil / 3Fundação de Medicina Tropical Doutor Heitor Vieira Dourado. Manaus. AM, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Transmissores de Hematozoários.. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Fisiologia e Controle de Artrópodes Vetores. Rio de Janeiro, RJ, Brasil / Instituto de Biologia do Exército. Laboratório de Entomologia. Rio de Janeiro, RJ, Brasil.Odebrecht Angola - Projectos e Serviços Ltda. Luanda, Angola.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Transmissores de Hematozoários.. Rio de Janeiro, RJ, Brasil.Distribution, abundance, feeding behaviour, host preference, parity status and human-biting and infection rates are among the medical entomological parameters evaluated when determining the vector capacity of mosquito species. To evaluate these parameters, mosquitoes must be collected using an appropriate method. Malaria is primarily transmitted by anthropophilic and synanthropic anophelines. Thus, collection methods must result in the identification of the anthropophilic species and efficiently evaluate the parameters involved in malaria transmission dynamics. Consequently, human landing catches would be the most appropriate method if not for their inherent risk. The choice of alternative anopheline collection methods, such as traps, must consider their effectiveness in reproducing the efficiency of human attraction. Collection methods lure mosquitoes by using a mixture of olfactory, visual and thermal cues. Here, we reviewed, classified and compared the efficiency of anopheline collection methods, with an emphasis on Neotropical anthropophilic species, especially Anopheles darlingi, in distinct malaria epidemiological conditions in Brazil