Occurrence of Bartonella genotypes in bats and associated Streblidae flies from Maranhão state, northeastern Brazil / Ocorrência de genótipos de Bartonella em morcegos e moscas Streblidae no Estado do Maranhão, Nordeste do Brasil

Abstract Bartonella is a genus of emerging zoonotic bacteria that are mainly associated with mammalian erythrocytes and endothelial cells. Bats are natural reservoirs for a variety of important pathogens that impact human and animal health. Recent reports have highlighted the role of bats and bat flies in the maintenance of Bartonella. Here, we showed that none of the 29 bat DNA blood samples obtained from five bat species in São Luís Island, state of Maranhão, northeastern Brazil, were positive for Bartonella in qPCR assays targeting nuoG. On the other hand, three out of 15 DNA samples (20%) from flies in the family Streblidae were positive for Bartonella. The BLASTn results showed that the gltA and rpoB sequences shared identities ranging from 97.2% to 100%, with Bartonella sequences amplified from bats or bat flies from Costa Rica and Brazil. These findings were supported by phylogenetic analyses based on Bayesian inferences. The present study showed that Bartonella genotypes are present in bat flies, thus shedding some light on the distribution of bat fly-related Bartonella genotypes in South America.

Resumo Bartonella é um gênero de bactérias zoonóticas emergentes associadas principalmente a eritrócitos e células endoteliais de mamíferos. Morcegos são reservatórios naturais para uma variedade de patógenos importantes que afetam a saúde humana e animal. Além disso, estudos recentes destacaram o papel dos morcegos e de moscas associadas a morcegos na manutenção de Bartonella. No presente estudo, nenhuma das 29 amostras de DNA obtidas a partir do sangue de cinco espécies de morcegos amostrados na ilha de São Luís, estado do Maranhão, Nordeste do Brasil, foi positiva para Bartonella nos ensaios de qPCR direcionados ao gene nuoG. Por outro lado, três das 15 (20%) amostras de DNA de moscas da família Streblidae foram positivas para Bartonella. Os resultados do BLASTn mostraram que as sequências dos genes gltA e rpoB compartilharam identidade, variando de 97,2% a 100%, com as sequências de Bartonella amplificadas em morcegos ou moscas amostrados na Costa Rica ou Brasil. Tais resultados corroboraram as análises filogenéticas realizadas por Inferência Bayesiana. O presente estudo mostrou a ocorrência de Bartonella em moscas de morcegos, auxiliando a esclarecer a distribuição dos genótipos de Bartonella relacionadas a moscas Streblidae na América do Sul.

Preliminary screening of the larvicidal effect of Brevibacillus laterosporus strains against the blowfly Chrysomya megacephala(Fabricius, 1794) (Diptera: Calliphoridae)

: This study evaluated whether different strains of Brevibacillus laterosporus could be used to control larvae of the blowfly Chrysomya megacephala , a pest that affects both human and animal health.

Molecular and parasitological detection of Leishmania spp. in a dipteran of the species Tabanus importunus / Detecção molecular e parasitológica de Leishmania spp. em díptero da espécie Tabanus importunus

Leishmaniasis is an important chronic zoonosis caused by protozoa of the genus Leishmania spp. The major vectors of this protozoosis are sand flies, and Lutzomyia longipalpis is considered the main species implicated in the transmission of American Visceral Leishmaniasis in Brazil. The presence of the parasite's deoxyribonucleic acid (DNA) in ectoparasites such as ticks and fleas has prompted speculations about the existence of new vectors in the cycle of leishmaniasis. The aim of this paper is to report the molecular detection of Leishmania spp. in a horse fly of the species Tabanus importunus which parasitized an oligosymptomatic dog infected with Leishmania spp. Molecular amplification of the protozoan's DNA in the head, thoracic region and abdomen of the tabanid tested positive for Leishmania complex. This is the first report of the presence of DNA from Leishmania spp. in dipterous insects of the species T. importunus.

A leishmaniose é uma importante zoonose, de caráter crônico, causada por protozoários do gênero Leishmania spp. Esta protozoose tem como principal vetor os flebotomíneos, sendo que, no Brasil, o Lutzomyia longipalpis é a principal espécie incriminada na transmissão da leishmaniose Visceral Americana. A presença do ácido desoxirribonucleico (DNA) do parasito em ectoparasitos, como carrapatos e pulgas, tem gerado especulações quanto a existência de novos vetores no ciclo da leishmaniose. Foi objetivo deste estudo relatar a detecção molecular de Leishmania spp. em uma mutuca da espécie Tabanus importunus que parasitava um cão oligossintomático infectado por Leishmania spp. A análise molecular amplificou o DNA do protozoário na cabeça, na região torácica e no abdomen do tabanídeo, resultando como positivo para complexo Leishmania. Este é o primeiro relato da presença de DNA de Leishmania spp. em insetos dipteros da espécie T. importunus.

Primer registro para el estado Bolívar de Culex (Lutzia) bigoti Bellardi, 1862 (Diptera: Culicidae) / First record of Culex (Lutzia) bigoti Bellardi, 1862 (Diptera: Culicidae) un Bolivar state, Venezuela

En el presente trabajo se reporta por primera vez para el estado Bolívar la presencia de Culex (Lutzia) bigoti Bellardi. Con este nuevo registro la distribución geográfica de esta especie en Venezuela incluye hasta ahora doce (12) estados y el Distrito Federal

In this work we report for the first time the presence of Culex (Lutzia) bigoti Bellardi in Bolivar state. With this new record the geographical distribution of this species in Venezuela include so far twelve (12) states and the Distrito Federal

New record of Pterotaenia fasciata (Wiedemann) (Diptera, Ulidiidae) in Brazil, a probably mechanical vector of enteric bacteria

Pterotaenia fasciata is commonly recorded in rural areas in Argentina, but during a Diptera survey study developed in a reservoir which retains storm water from polluted canals in an urban area of Taboão da Serra municipality, SP, Brazil, we could capture P. fasciata adults. Enteric bacteria Escherichia coli T. Escherich, 1885 and Proteus sp. were isolated from P. fasciata collected in traps inside the reservoir and around it. Fecal coliforms and E. coli were found in the water of the reservoir. These records suggest that a high abundance of this species at urban areas with inadequate sewage canals should reveal these muscoid dipterans as mechanical vectors of enteric bacteria.

Pterotaenia fasciata é encontrada freqüentemente em áreas rurais na Argentina, mas durante um estudo de levantamento de Diptera em um reservatório de retenção de enchentes em uma área urbana do município de Taboão da Serra, SP, Brasil, foram capturados adultos de P. fasciata. As enterobactérias Escherichia coli T. Escherich, 1885 e Proteus sp. foram isoladas de P. fasciata coletada em armadilhas no reservatório e em seu entorno. Coliformes fecais e E. coli foram encontrados na água do reservatório. Esses registros sugerem que a alta abundância dessa espécie em áreas urbanas sem saneamento básico poderia indicar esses dípteros muscóides como vetores mecânicos de enterobactérias.

Comparison between Musca domestica and Chrysomya megacephala as carriers of bacteria in northern Thailand.

A comparison between the common house fly, Musca domestica, and the Oriental latrine fly, Chrysomya megacephala, was assessed for their potential as carriers of bacteria in urban areas of Chiang Mai Province, northern Thailand. C. megacephala was significantly more likely to carry bacterial species than M. domestica; however, no significant difference was found between the number of positive male and female flies within the same species. A total of 42 bacterial species were isolated. The most common bacterium isolated from M. domestica was coagulase-negative staphylococci (n=57) followed by Escherichia coli (n=10) and Viridans streptococci (n=10), while that of C. megacephala was non-fermentative gram-negative bacilli (n=59) followed by coagulase-negative staphylococci (n=54).

Biological activity of Bacillus thuringiensis strains against larvae of the Blowfly Chrysomya putoria (Wiedemann) (Diptera: Calliphoridae)

Different strains of Bacillus thuringiensis Berliner were proved to be a powerful biologic insecticide against larvae of several insect orders. Due to the epidemiological importance of blowflies of the Chrysomya Robineau-Desvoidy genus in the production of secondary cutaneous myiasis and mechanic transmission of pathogenic agents, the performance of two strains of B. thuringiensis (LFB-FIOCRUZ 907 and LFB-FIOCRUZ 856) was tested against larvae of Chrysomya putoria (Wiedemann). The LFB-FIOCRUZ 907 strain was tested in four different concentrations, added to the diet; the LFB-FIOCRUZ 856 strain was tested in three concentrations. C. putoria larvae showed sensibility to the treatment with the LFB-FIOCRUZ 907 strain at the tested concentrations. The higher concentration presented the best efficiency, causing higher mortality and reducing larval weight and adult emergence more intensely. The LFB-FIOCRUZ 856 strain showed low toxicity, sliglity reducing emergence time of adults at 326 mg/ 25 g concentration and larval weight at 326 mg/ 25 g and 86 mg/ 25 g concentrations.

Diferentes linhagens de Bacillus thuringiensis Berliner têm provado ser poderoso inseticida biológico contra larvas de várias ordens de insetos. Dada a importância epidemiológica das moscas do gênero Chrysomya Robineau-Desvoidy na produção de miíases cutâneas secundárias e transmissão mecânica de agentes patogênicos, avaliou-se a atividade de duas linhagens de B. thuringiensis (LFB-FIOCRUZ 907 e LFB-FIOCRUZ 856) sobre larvas de Chrysomya putoria (Wiedemann). A linhagem LFB-FIOCRUZ 907 foi testada em quatro diferentes concentrações misturadas à dieta e a linhagem LFB-FIOCRUZ 856 em três concentrações. As larvas de C. putoria apresentaram sensibilidade ao tratamento com a linhagem LFB-FIOCRUZ 907, nas concentrações testadas, sendo a concentração mais alta a de maior eficiência, causando maior mortalidade e reduzindo mais intensamente o peso larval e a taxa de emergência dos adultos. A linhagem LFB-FIOCRUZ 856 apresentou toxicidade muito baixa, redizindo ligeiramente a emergência dos adultos na concentração de 326 mg/ 25 g e o peso larval nas concentrações de 326 mg/ 25 g e 86 mg/ 25 g.

Biologia e exigências térmicas de Trichogramma pratissolii Querino & Zucchi (Hymenoptera: Trichogrammatidae) em hospedeiros alternativos / Biology and thermal requirements Trichogramma pratissolii Querino & Zucchi (Hymenoptera: Trichogrammatidae) on alternative hosts

O objetivo deste trabalho foi estudar a biologia e as exigências térmicas de Trichogramma pratissolii Querino & Zucchi em Anagasta kuehniella Zeller (Lepidoptera: Pyralidae) e Corcyra cephalonica Stainton (Lepidoptera: Pyralidae), nas temperaturas de 15, 18, 21, 24, 27, 30 e 33°C. A porcentagem de emergência de T. pratissolii em A. kuehniella foi maior a 27°C, e em C. cephalonica, foi maior às temperaturas de 24°C a 30°C. O limite térmico inferior de desenvolvimento (Tb) e a constante térmica (K) de T. pratissolii foram 12,59°C (Tb) e 122,85 graus-dias (K) em A. kuehniella e 11,73°C (Tb) e 139,80 graus-dias (K) em C. cephalonica. Os resultados indicam que ambos hospedeiros A. kuehniella e C. cephalonica podem ser utilizados para criação massal de T. pratissolii. A melhor temperatura para desenvolvimento da forma imatura e para a emergência de T. pratissolii em A. kuehniella foi 27°C e, em C. cephalonica, na faixa de 24°C a 30°C.

The objective of this work was to study the biology and determine the thermal requirements of Trichogramma pratissolii Querino & Zucchi reared on Anagasta kuehniella Zeller (Lepidoptera: Pyralidae) and Corcyra cephalonica Stainton (Lepidoptera: Pyralidae) at 15, 18, 21, 24, 27, 30 and 33°C. The highest percentage of emergence of T. pratissolii was obtained at 27°C on A. kuehniella and from 24°C to 30°C on C. cephalonica. The lower threshold temperature (Tb) and the thermal requirement (K) of T. pratissolii were 12.59°C (Tb) and 122.85 degree-days (K) when reared on A. kuehniella and 11.73°C (Tb) and 139.80 degree-days (K) on C. cephalonica. These results indicate that both A. kuehniella and C. cephalonica are suitable hosts for T. pratissolii mass rearing. The optimal temperature for the immature development and emergence of T. pratissolii on A. kuehniella was 27°C and 24°C to 30°C on C. cephalonica.

Pathogenicity of Metarhizium anisopliae for Ceratitis capitata (Wied. ) (Diptera: Tephritidae) in soil with different pesticides

This research intended to investigate if the presence of pesticides in the soil could affect the pathogenicity of Metarhizium anisopliae Metsch. (Sorokin) for Ceratitis capitata (Wied.) and assess the effect of conidia application as suspension or dry conidia. The fungicides chlorothalonyl and tebuconazol, the acaricide abamectin, the insecticide trichlorfon, and the herbicide ametrin were applied at the manufacturer-recommended doses. Soil samples were placed in glass flasks and were given the fungus as conidial suspension or dry. After pesticide application, 20 3rd-instar larvae were placed in the soil. The flasks were sealed with voile fabric and incubated at 27 ± 0.5°C for nine days, until adult emergence; incubation continued for four more days at room temperature. The total insect survival was significantly affected and pathogenic activity was detected from the pupa stage on. Pupa survival was reduced (P<0.05); the same occurred during the adult phase. No effect was observed at the larval stage. The pesticides applied to the soil affected the activity of M. anisopliae slightly: only in the dry conidia assay the fungicides chlorothalonyl and tebuconazole reduced (86.2 percent and 82.5 percent, respectively) the survival period of C. capitata compared to the control (95.0 percent). The techniques used for conidia application did not influence the total insect survival rate, but conidial suspension applied on soil surface reduced survival during the pupae and adult phases.

O presente trabalho objetivou investigar se a presença de agrotóxicos no solo afeta a patogenicidade de Metarhizium anisopliae Metsch. (Sorokin) para Ceratitis capitata (Wied.). Avaliou-se também o efeito das formas de aplicação dos conídios. Foram utilizados os fungicidas clorotalonil e tebuconazole, o acaricida abamectina, o inseticida triclorfom e o herbicida ametrina, aplicados nas doses recomendadas pelos fabricantes. Porções de solo contidas em frascos de vidro receberam o fungo na forma de suspensão de conídios ou como conídios secos incorporados. Após a aplicação dos agrotóxicos, 20 larvas de 3° ínstar foram colocadas no solo. Os frascos foram então vedados com tecido voile e incubados a 27 ± 0,5°C por nove dias, para iniciar a emergência dos adultos, e mais quatro dias à temperatura ambiente. A sobrevivência total do inseto foi substancialmente afetada. Observou-se a atividade patogênica a partir da fase pupal, com o fungo reduzindo (P < 0,05) a sobrevivência, o que também ocorreu na fase de adultos. A fase larval não foi afetada (P >0,05) pelo fungo. A presença de agrotóxicos no solo teve discreta ação na atividade de M. anisopliae; apenas com conídios secos incorporados, os fungicidas clorotalonil e tebuconazole reduziram em 86,2 por cento e 82,5 por cento, respectivamente, a sobrevivência total de C. capitata, enquanto no controle a redução foi de 95,0 por cento. A forma de aplicação dos conídios não influenciou a sobrevivência total do inseto, mas a aplicação da suspensão de conídios na superfície do solo reduziu a sobrevivência nas fases de pupa e adulto.

Effect of microwave irradiation on the blow fly Chrysomya megacephala (F.) (Diptera: Calliphoridae).

Chrysomya megacephala (F) is a blow fly species of medical importance, being pestiferous and a mechanical carrier of various microorganisms to human food. Various strategies to reduce the fly population in endemic areas are needed. We investigated the effects of microwaves at 2,450 MHz, with third-instar exposure times of 7, 10, 15, 30 and 60 seconds on the larvae. Survival of the larvae was monitored by their emergence. About 60% of the larvae lived after 7 seconds of exposure, most died by 15 seconds, and all were dead at 30 and 60 seconds. The number of survivals decreased with increasing exposure times. This indicates the effect of microwave irradiation on fly larval survival.

Karyotype characterization of Anastrepha fruit flies (Diptera: Tephritidae)

The study comprises the description of the C-banded mitotic chromosomes of five species of Anastrepha, A. amita Zucchi, A. turpiniae Stone, A. zenildae Zucchi, A. grandis (Macquart) and A. leptozona Hendel , and a reanalysis of the chromosomes of three other species, A. distincta Greene, A. obliqua (Macquart) and A. sororcula Zucchi. The species have a diploid number of 12, while A. leptozona has 2n = 10. Heteromorphism of the sex chromosomes is present in all of the species, which have a XY/XX sex chromosome system. A. grandis and A. leptozona have meta and submetacentric autosomes while all of the chromosomes in the other species are acrocentric. The species can be characterized unequivocally by the length and the pattern of blocks of heterochromatin in the sex chromosomes.

O estudo apresenta a descrição dos cromossomos mitóticos, através do bandamento-C, de cinco espécies de Anastrepha, A. amita Zucchi, A. turpiniae Stone, A. zenildae Zucchi, A. grandis (Macquart) and A. leptozona Hendel e uma reanálise dos carótipos de três outras espécies, A. distincta Greene, A. obliqua (Macquart) and A. sororcula Zucchi, segundo a mesma técnica. As espécies apresentam número diplóide igual a 12 cromossomos, exceto A. leptozona com 2n = 10. Os cromossomos sexuais são heteromórficos e com sistema XX/XY. Os autossomos são meta ou submetacêntricos em A. grandis and A. leptozona e acrocêntricos nas demais espécies. Os cromossomos sexuais apresentam comprimento e padrão de blocos de heterocromatina característicos para cada espécie.

Defense reactions of Dermatobia hominis (Diptera: Cuterebridae) larval hemocytes

The defense reactions against biological (Histoplasma capsulatum and Escherichia coli) and non-biological materials (China ink and nylon thread) were tested in vivo in third instar larvae of Dermatobia hominis. The cellular defense performed by larval hemocytes was observed under electron microscopy. China ink particles were phagocytosed by granular cells 5 h after injection. E. coli cells were internalized by granular cells as early as 5 min after injection and totally cleared 180 min post-injection, when many hemocytes appeared disintegrated and others in process of recovering. H. capsulatum yeasts provoked, 24 h after being injected, the beginning of nodule formation. Nylon thread was encapsulated 24 h after the introduction into the hemocoel. Our results suggest that granular cells were the phagocytic cells and also the responsible for the triggering of nodule and capsule formation. In the presence of yeasts cells and nylon thread, they released their granules that chemotactically attracted the plasmatocytes that on their turn, flattened to surround and isolate the foreign material.

Mechanical carrier of bacterial enteric pathogens by Chrysomya megacephala (Diptera: Calliphoridae) in Chiang Mai, Thailand.

Chrysomya megacephala was studied regarding its mechanically bacterial carrier in urban areas of Chiang Mai, northern Thailand. Fifty-six adult flies were randomly collected using sweep insect net during April-May, 1999 from 3 fresh food markets and examined for bacteriological isolation. Among them, 49 flies (87.5%) were bacterial carriers. The total 22 bacterial species and 8 groups were isolated. Three species previously reported as the bacterial enteric pathogens causing diarrheal disease were isolated from 5 flies, ie Aeromonas hydrophila, Edwardsiella tarda and Vibrio cholerae non-01, with their prevalence rates in flies being 3.579, 1.79% and 3.57%, respectively. Five possible bacteria enteric pathogens, ie Aermononas sobria, Citrobacter freundii, Escherichia coli, Providencia alcalifaciens and Pseudomonas aeruginosa, were isolated from 21 flies with the prevalence rates in flies being 5.34%, 3.57%, 26.79%, 7.14% and 1.79%, respectively. The bacterial load isolated from all 3 pathogenic species was entirely found more than 10 colony per fly, indicating the high chance for disease transmission via this fly species. C. megacephala may play the possible and/or important role of bacterial enteric pathogens transmission, thereby promoting the public health personnel for sanitation improvement in fresh food markets and fly control management in these particular areas.

Presencia de bacterias patógenas en las moscas / Presence of pathogenic bacteria in flies

Se lleva a efecto una investigación microbiológica en una muestra de la población de moscas en un área de salud del municipio Marianao. Se emplean selectivos para la búsqueda de enterobacterias patógenas, y se demuestra la presencia de contaminación en el 100% de las mismas. Se observa el predominio de la Pseudomona aeruginosa (87,5%) sobre estos gérmenes entéricos gramnegativos, también presentes. Se demuestra el carácter de vector de estos artrópodos, y la variedad de agentes que transportan

Disease agents carried by flies in Dacca city.

Studies were conducted to determine the different disease agents naturally carried by flies in Dacca city by collecting outdoor unbaited flies from 8 different representative locations and identifying the agents in the laboratory. Out of 96 fly pools containing 25 flies each, 62.5%, 46.9%, 15.6%, 8.3%, 8.3%, 15.6% and 6.2% of the pools were found positive for Ascaris lumbrioeides, Trichuris trichiura, Hookworm, Enterebious vermicularis eggs and Entamoeba coli, Trichomonas spp Giardia spp. cysts, respectively. Similarly, out of the same number of fly pools, 8.3%, 6.25%, 66.6%, 20.8% and 29.2% of the pools were found positive for Shigella flexner-3, Cholera vibrio, Escherichia coli, Staphylococcus aureus and Pseudomonas, respectively besides many other non-pathogenic bacteria.