Techniques To Improve The Maintenance Of A Laboratory Colony Of Nyssomyia Neivai (diptera: Psychodidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Background: The most critical phase in sand fly colonization is the high mortality in the larval instars. In this study, we sought out strategies for improving the colonization of Nyssomyia neivai, one of the vectors of cutaneous leishmaniasis agent in South America. Methods: A colony of Ny. neivai was established in the laboratory from a field population, and the productivity of adults was evaluated considering carrying capacity, diet for larvae and surface for oviposition. Results: The highest emergency rate of adults was achieved with the fewest couples inside 150 mL rearing chambers on a sterilized diet made of rabbit feces, rabbit food, soil and fish food and with vermiculite as a substrate for oviposition and the development of larvae. Conclusion: Our data on Ny. neivai colonization showed that the best adult productivities were achieved with fewer couples inside the rearing chambers; smaller rearing containers of 150 mL (due to less fungi growth); sterilized diet made of rabbit feces, rabbit food, soil and fish food; and vermiculite as the substrate for oviposition and development of larvae.8Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Programa de Apoio ao Desenvolvimento Cientifico da Faculdade de Ciencias Farmaceuticas at UNESP (PADC/FCFAr)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP [2012/23832-9

Attraction Of The Sand Fly Nyssomyia Neivai (diptera: Psychodidae) To Chemical Compounds In A Wind Tunnel.

Similar to other hematophagous insects, male and female sand flies must feed on plants to obtain sugar and, subsequently, energy to complete their life cycles. A large number of compounds emitted by plants may act as volatile signals to these insects. Primary alcohols have been detected in some plants, but in small amounts. In a previous report, the attractiveness of saturated primary alcohols with 7 to 9 carbons was evaluated for Lutzomyia longipalpis, the vector of American visceral leishmaniasis, with positive results. In the present study, a wide range of primary alcohols, 3 to 10 carbons, were tested to investigate their attractiveness to another sand fly species, Nyssomyia neivai, a putative vector of American cutaneous leishmaniasis. The mixture of compounds that induced the best sand fly response was also evaluated. Of the eight compounds evaluated, hexanol and octanol elicited the best attractive responses for sand fly females. Phytochemicals may be an interesting source of search for new sand fly attractants.814

Attraction of the sand fly Nyssomyia neivai (Diptera: Psychodidae) to chemical compounds in a wind tunnel

Similar to other hematophagous insects, male and female sand flies must feed on plants to obtain sugar and, subsequently, energy to complete their life cycles. A large number of compounds emitted by plants may act as volatile signals to these insects. Primary alcohols have been detected in some plants, but in small amounts. In a previous report, the attractiveness of saturated primary alcohols with 7 to 9 carbons was evaluated for Lutzomyia longipalpis, the vector of American visceral leishmaniasis, with positive results. In the present study, a wide range of primary alcohols, 3 to 10 carbons, were tested to investigate their attractiveness to another sand fly species, Nyssomyia neivai, a putative vector of American cutaneous leishmaniasis. The mixture of compounds that induced the best sand fly response was also evaluated. Of the eight compounds evaluated, hexanol and octanol elicited the best attractive responses for sand fly females.Phytochemicals may be an interesting source of search for new sand fly attractant

Mites associated with the eared dove Zenaida auriculata (Des Murs, 1847), in São Paulo State, Brazil

Orientador: Angelo Pires do PradoDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: Este trabalho objetivou o levantamento e a complementação das poucas informações sobre as espécies de ácaros associados à Zenaida auriculata na região de Campinas, SP [(Campinas (22°49'11''S, 47°4'12''O, alt.604m), Valinhos (22°58'14"S, 47°59'45"O, alt. 660m), Jaguariúna (22° 42' 20''S, 46° 59' 09''O, alt. 584m), Santa Bárbara D'oeste (22°45'13"S, 47°24'49"O, alt. 565m)] e em outras localidades como Ourinhos (22°58'44"S, 49°52'14"O, alt. 483m). No total foram analisadas 54 amostras, sendo que em 10 delas, não foram encontrados ácaros. Como resultado, obtivemos representantes das seguintes subordens: Astigmata: Falculiferidae - Falculifer isodontus Gaud & Barré, 1992; Byersalges talpacoti Cerný, 1975; Pterophagus spilosikyus Gaud & Barré, 1992; Dermoglyphidae - Dermoglyphus columbae Sugimoto, 1941; Analgidae: Diplaegidia columbae Buchholz, 1869; Diplaegidia columbigallinae Cerný, 1975; Epidermoptidae não identificado; Pyroglyphidae não identificado; Hypoderatidae: Hypodectes propus (Nitzch, 1861); Prostigmata - Cheyletiellidae: Ornithocheyletia columbigallinae Fain & Bochkov, 2002; Cheyletidae não identificado; Syringophilidae: Meitingsunes zenadourae Clark 1964; Mesostigmata - Macronyssidae: Ornithonyssus bursa (Berlese, 1888); Rhinonyssidae: Tinaminyssus zenaidurae (Crossley, 1952). As ocorrências de H. propus (deutoninfas parasitas subcutâneas), B. talpacoti, T. zenaidurae, P. spilosikyus, O. columbigallinae, O. bursa, Dermoglyphus columbae são relatadas pela primeira vez nos adultos deste hospedeiro. Foram encontradas 14 spp. de ácaros, distribuídas em: Astigmata 9, Prostigmata 3 e Mesostigmata 2. Sendo 1 sp.do aparelho respiratório; 4 spp. da pele; 2 spp nidícolas, uma delas hematófaga; 2 spp. calamícolas; 5 spp. plumícolas. O material testemunho foi depositado na coleção do Departamento de Biologia Animal, Instituto de Biologia, UNICAMP, além das fotografias dos ninhos, das aves e dos ácaros (coleção virtual)Abstract: This study aimed to add information on the species of mites associated with Zenaida auriculata in the region of Campinas, SP. [(Campinas (22°49'11''S, 47°4'12''W, alt.604m), Valinhos (22°58'14"S, 47°59'45"W, alt. 660m), Jaguariúna (22° 42' 20''S, 46° 59' 09''W, alt. 584m), Santa Bárbara D'oeste (22°45'13"S, 47°24'49"W, alt. 565m)] and in Ourinhos (22°58'44"S, 49°52'14"O, alt. 483m). A total of 54 samples were examined, and mites were found on 44 of them. We found representatives from the following taxa: Astigmata: Falculiferidae - Falculifer isodontus Gaud & Barré, 1992, Byersalges talpacoti Cerný, 1975; Pterophagus spilosikyus Gaud & Barré, 1992; Dermoglyphidae - Dermoglyphus columbae Sugimoto, 1941; Analgidae: Diplaegidia columbae Buchholz, 1869; Diplaegidia columbigallinae Cerný, 1975; an unidentified Epidermoptidae; an unidentified Pyroglyphidae; Hypoderatidae: Hypodectes propus (Nitzch, 1861); Prostigmata - Cheyletiellidae: Ornithocheyletia columbigallinae Fain & Bochkov 2002; an unidentified Cheyletidae; Syringophilidae Meitingsunes zenadourae (Clark, 1964); Mesostigmata - Macronyssidae: Ornithonyssus bursa (Berlese, 1888); Rhinonyssidae: Tinaminyssus zenaidurae (Crossley, 1952). The occurrence of Hypodectes propus (subcutaneous deutonymphs parasites), B. talpacoti, T. zenaidurae, P. spilosikyus, O. columbigallinae, O. bursa and Dermoglyphus columbae are reported for the first time in the adult form of this host. Fourteen species of mites were found, distributed as follow: Astigmata 9, Prostigmata 3 e Mesostigmata 2. One species was of respiratory system; four of skin; two was nidiculous, but only one was hematophagous; two of quill mites; five were feather mites. Voucher specimens were deposited at a collection of Department of Animal Biology, Biology Institute, UNICAMP, in addition to photographs of nests, birds and mites (virtual collection)MestradoParasitologiaMestre em Parasitologi

Study of biological aspects of Nyssomyia neivai (Diptera: Psychodidae) and improvement of colonization techniques in laboratory

Orientadores: Mara Cristina Pinto, Marili Villa Nova RodriguesTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: Algumas espécies de flebotomíneos (Diptera: Psychodidae), transmissoras de leishmania, são de difícil manutenção ou baixa produtividade em laboratório. Essas limitações prejudicam a realização de bioensaios que, de modo geral, utilizam grande quantidade de insetos. As tentativas de manutenção da espécie Nyssomyia neivai (=Lutzomyia neivai), considerada de importância na transmissão da leishmaniose tegumentar Americana, em laboratório são escassas, além de serem observadas altas taxas de mortalidade quando utilizados protocolos para outras espécies de flebotomíneos. Visando aumentar os conhecimentos de biologia básica de Ny. neivai e o aprimoramento de técnicas de criação mais específicas e eficientes, este estudo teve como objetivo principal investigar alternativas possíveis durante as diferentes fases de desenvolvimento do inseto. Basicamente quatro pontos cruciais no processo de criação foram focados: fonte alimentar sanguínea para as fêmeas, fonte alimentar de açúcares para os adultos, dieta das formas imaturas e investigação da presença de feromônios de oviposição. Entretanto, ao longo do processo de colonização de Ny. neivai, e das dificuldades encontradas, observamos outros pontos que poderiam apresentar relevância para um melhor rendimento no número final de adultos produzidos. Como resultado, foi obtida a maior produtividade de adultos e menor perda de larvas adicionando de cinco a dez casais em recipientes de criação (145 mL) para oviposição. A vermiculita mostrou-se um ótimo substrato a ser adicionado por cima do gesso na criação das larvas. A dieta artificial tradicional autoclavada, mostrou-se mais atrativa dentre as testadas e proporcionou o melhor desenvolvimento das formas imaturas. O camundongo mostrou-se a melhor alternativa como fonte alimentar sanguínea para os insetos. A alimentação artificial com membrana não foi eficiente para a manutenção da colônia, porém, para estudos de infecção de poucos indivíduos por micro-organismos, a alimentação artificial com membrana de pele de camundongo e sangue humano pode ser uma alternativa viável. Através da cromatografia gasosa, foram encontrados os carboidratos: frutose, glucose, isomaltose, ribose e trealose em insetos de campo e foi avaliada a curva de sobrevivência dos insetos alimentados com os três primeiros açúcares, separadamente, e com dois alimentos glicosados comerciais, resultando na melhor opção para a rotina laboratorial o alimento glicosado Favinho®. Na procura de feromônios de oviposição, experimentos de evidência biológica foram delineados e em todos esses experimentos não houve indicação da presença de voláteis na superfície dos ovos. Ainda na tentativa de isolamento químico de compostos nos ovos de Ny. neivai testes foram realizados com Microextração em fase sólida (SPME) e GC-MS. Esses testes apresentaram contaminantes nos cromatogramas, impedindo sua real análise e encobrindo possíveis picos de interesse. A partir desses resultados levanta-se a hipótese de que Ny. neivai não possua feromônios de oviposição, apresentando resposta visual. Porém, pode haver um sinergismo entre a resposta visual e os feromônios ainda não detectados pela metodologia adotadaAbstract: Some sandflies species (Diptera: Psychodidae), vector of leishmania are difficult to maintain or to produce in laboratory conditions. These limitations prejudice experiments that demand high number of insects. Nyssomyia neivai (=Lutzomyia neivai) is considered important in the transmission of the American Cutaneous Leishmaniasis and it is a specie with the features above. The attempts to maintain this specie in laboratory conditions are scarce and follow another phlebotomine species protocol resulting in a great mortality of insects. Aiming to increase the knowledge of the basic biology of Ny. neivai and the improvement of the rearing techniques to be more specific and efficient, the main purpose of this project was to investigate possible alternatives during the different development phases of the insect. Basically, four crucial points were focused: blood source for female sandflies, sugar source for adults, diet for immature stages and investigation of presence of oviposition pheromones. However, in this long process of colonization of Ny. neivai and the difficulties found, we were able to observe another points that could be relevant for a higher number of produced adults. The best result was achieved with five to ten couples inside a rearing pot (145 mL) for oviposition and vermiculite grains proved to be a great substrate to cover the plaster of Paris for the larvae rearing. The traditional diet, sterilized by autoclave, proved to be more attractive among the diets tested and provided better development for immature stages. The mice proved to be the best blood source for the female insects. The membrane feeding technique was not efficient to maintain the colony; however, it is a viable method for studies, with few sandflies, of infection with microorganism using mice skin as membrane and human blood as blood source. Fructose, glucose, isomaltose, ribose and trealose were the carbohydrates found in field insects, using gas chromatography, and the survival curve of sandflies was analyzed with the three first sugars mentioned above; two more commercial products were offered to the insects. As result, the commercial product Favinho® proved to be the better one to be used in the laboratory routine. Searching for oviposition pheromones, experiments of biological evidence were performed and the results revealed no indication of volatiles in the surface of the eggs. In the attempt to isolate chemical compounds, Ny. neivai eggs were exposed to Solid Phase Microextraction (SPME) and GC-MS. The tests result presented contamination in the chromatograms, preventing the real analysis and occulting possible peaks of interest. Thus, these results raise the hypothesis that there is no oviposition pheromones in Ny. neivai, presenting visual stimulus. Nevertheless, it could be a synergism between visual stimulus and pheromones not yet detected with the methodology adoptedDoutoradoRelações Antrópicas, Meio Ambiente e ParasitologiaDoutora em Biologia AnimalCAPE

Solid Phase Microextraction, Sand Flies, Oviposition Pheromones, Plaster Of Paris And Siloxanes What Is In Common?

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Sand flies are natural hosts of various microorganisms. Due to their epidemiological importance, sand fly colonies are kept in laboratories to be studied in terms of their biology and vector/host/parasite interactions. In order to investigate the presence of oviposition pheromones in Nyssomyia neivai, experiments using Solid Phase Microextraction (SPME) were performed. However, siloxanes which is an external class of contamination, present in breeding containers made by plaster used to maintain sand flies in colonies, may be hindered the experiments. Published by Elsevier B.V.1685053FAPESP (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo-Process) [2012/23832-9]Programa de Apoio ao Desenvolvimento Cientifico da Faculdade de Ciencias Farmaceuticas at UNESP (PADC/FCFAr)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Techniques To Improve The Maintenance Of A Laboratory Colony Of Nyssomyia Neivai (diptera: Psychodidae).

The most critical phase in sand fly colonization is the high mortality in the larval instars. In this study, we sought out strategies for improving the colonization of Nyssomyia neivai, one of the vectors of cutaneous leishmaniasis agent in South America. A colony of Ny. neivai was established in the laboratory from a field population, and the productivity of adults was evaluated considering carrying capacity, diet for larvae and surface for oviposition. The highest emergency rate of adults was achieved with the fewest couples inside 150 mL rearing chambers on a sterilized diet made of rabbit feces, rabbit food, soil and fish food and with vermiculite as a substrate for oviposition and the development of larvae. Our data on Ny. neivai colonization showed that the best adult productivities were achieved with fewer couples inside the rearing chambers; smaller rearing containers of 150 mL (due to less fungi growth); sterilized diet made of rabbit feces, rabbit food, soil and fish food; and vermiculite as the substrate for oviposition and development of larvae

Molecular detection of the blood meal source of sand flies (diptera: Psychodidae) in a transmission area of american cutaneous leishmaniasis, Parana state, Brazil

The feeding behavior of sand flies provides valuable information about the vector/host interactions and elucidates the epidemiological patterns of American cutaneous leishmaniasis (ACL) transmission. The aim of this study was to identify the blood meal sources of sand flies in endemic areas of leishmaniasis in Parana State through polymerase chain reaction (PCR) amplification of a prepronociceptin (PNOC) gene fragment and its subsequent DNA sequencing. Moreover, molecular assays were conducted to evaluate the sensitivity and reproducibility of the PNOC gene amplification. Besides that, a time-course digestion test of the blood using sand flies that fed artificially on BALB/c mice was performed. Of 1263 female sand flies collected in the field, 93 (3.6%) specimens were engorged and 27 allowed efficient amplification of the PNOC gene. These flies had fed on equine (Equus caballus), porcine (Sus scrofa) and canine (Canis lupus familiaris) species. The results also showed that the identification of the blood meal sources of the sand flies using the molecular method was directly linked to the level of digestion of the blood (time-course) and not to the amount of blood that had been ingested or to the presence of inhibitors in the blood143812CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPE