Hemilucilia segmentaria (Fabricius, 1805) (Diptera: Calliphoridae) as new biological vector of eggs of Dermatobia hominis (Linnaeus Jr., 1781) (Diptera: Oestridae) in Reserva Biológica do Tinguá, Rio de Janeiro, Brazil

The aim of this note was to record for the first time the finding of Hemilucilia segmentaria acting as biological vector of Dermatobia hominis, during a study of the diversity of Calliphoridae at Reserva Biológica do Tinguá, Rio de Janeiro, Brazil. The insects were captured using traps baited with chicken vicera, for a period of 28-30 h twice per month. In the period of one year, 1987 insects were collected, 7.5% of which belonged to the H. segmentaria; of these a female was captured in May 2001, carrying a mass of 20 eggs on the left side of its abdomen.

Flutuação sazonal de Dermatobia hominis em peles bovinas oriundas de matadouro

Realizou-se o estudo do efeito da temperatura, precipitação pluviométrica e da umidade relativa sobre a flutuação sazonal de larvas de Dermatobia hominis em 5.142 peles de bovinos. As peles foram obtidas junto a um matadouro localizado no município de Piraí, estado do Rio de Janeiro, Brasil, durante o período de novembro de 1995 a outubro de 1997. Os maiores picos de peles parasitadas por berne foram observados em novembro de 1995 e dezembro de 1996. Picos menores de infestação foram observados nos meses de março e maio de 1996 e março de 1997, quando também foram observadas altas porcentagens de peles infestadas. O aumento na porcentagem de peles parasitadas acompanhou o aumento da temperatura média mensal, sendo que temperaturas iguais ou maiores que 25ºC se mostraram benéficas ao aumento na porcentagem de peles parasitadas, assim como a precipitação pluviométrica, onde observou-se que períodos de chuvas mensais acima de 200 mm³ favoreceram ao aumento da porcentagem de peles parasitadas. As menores porcentagens de peles parasitadas por berne foram observadas durante o período de junho a outubro de 1996 e de 1997, quando verificou-se temperaturas médias mensais variando entre 21ºC e 22,5ºC, sendo que estas não se mostraram favoráveis ao aumento na porcentagem de peles parasitadas, assim como, precipitações pluviométricas mensais menores que 50 mm³. Das 5.142 peles examinadas, 2.710 (52,7%) encontravam-se parasitadas pelo berne.

Dípteros fanídeos vetores de ovos de Dermatobia hominis em Campo Grande, Mato Grosso do Sul

Determinou-se a importância epidemiológica de dípteros Fanniidae na infestação de mosca-do-berne, por meio da identificação das espécies presentes, da determinação daquelas utilizadas por Dermatobia hominis na veiculação de seus ovos, bem como, pelo conhecimento da dinâmica populacional das espécies mais abundantes. Foram utilizadas cinco armadilhas iscadas com fígado bovino cru deteriorado e colocadas em uma mata ciliar margeada por uma área de pastagem com presença constante de bovinos. O estudo foi desenvolvido em uma área da Embrapa Gado de Corte, em Campo Grande, Estado de Mato Grosso do Sul, Brasil, localizada a 20º27'S e 54º37'W. A captura dos insetos foi realizada semanalmente durante o período de 09/08/1999 a 03/08/2000. Foi capturado um total de 40.629 moscas da família Fanniidae, pertencendo a cinco espécies do gênero Fannia: F. pusio, F. heydenii, F. bahiensis e F. longipila, e uma a ser identificada. A espécie mais freqüente foi F. pusio, com 63,20% do total capturado, seguida de F. heydenii, com 28,82%. Somente 0,44% do total de fêmeas de F. heydenii (45 exemplares) capturadas, principalmente nos meses de agosto e setembro, portavam ovos de D. hominis e o número médio, por indivíduo, foi de 15,98±7,13. Observaram-se ovos de D. hominis apenas na região abdominal dos vetores. F. heydenii predominou no período seco (maio a setembro) e início do período chuvoso do ano (outubro e novembro). O número de exemplares portando ovos de D. hominis foi maior no final do período seco do ano, o que explica a alta incidência deste parasito em bovinos nos meses de setembro e outubro.

Inflammatory reaction to the human bot-fly, Dermatobia hominis, in infested and reinfested mice

Two groups of mice were infested with first stage larvae of the human bot-fly, Dermatobia hominis (Linnaeus Jr) (Diptera: Oestridae). In the first group, skin biopsies were carried out 1, 3, 5, 7, 10 and 18 days after infestation. The second group was also infested but had all the larvae removed 5 days after infestation. The mice in the latter group were reinfested 4 weeks later and skin biopsies were carried out 1, 3, 5, 7, 10 and 18 days after reinfestation. In the first group, an inflammatory reaction began slowly, the neutrophils being the main inflammatory cells, eosinophils being scarce. The reaction progressed with time, developing a necrotic halo around the larvae containing inflammatory cells surrounded by fibroblasts. The inflammation invaded the adjacent tissue. In the second group, the inflammatory reaction was intense on the day immediately after reinfestation, the pattern being changed by the presence of a large number of eosinophils. Activated fibroblasts surrounding the necrotic area around the larvae appeared 3 days after reinfestation in the second group and 7 days after infestation in the first group. The results demonstrated that the previous contact with the antigens elicited the early arrival of eosinophils, probably through the chemotactic factors liberated by mast cells in the anaphylactic reaction.

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.

Elementos figurados da hemolinfa de Dermatobia hominis (Diptera: Cuterebridae): caracterização ao nível de microscopia óptica, em larvas do 2o. e 3o. instares

Foram examinados os hemócitos de larvas do 2º (L2) e 3º(L3) instares de Dermatobia hominis em nível de microscopia óptica e comparados com os de outras espécies encontradas na literatura. Nas L2 e em L3 com peso de até 200mg foram encontrados cinco tipos: Pro-hemócitos, Plasmatócitos, Vermiformes, Oenocitóides e Esfoliativas. A medida em que as L3 foram-se tornando mais idosas apareceram em seqüência os Granulócitos e Adipohemócitos, sendo raro encontrar-se Pro-hemócitos em L3 com peso acima de 500mg. Tipos intermediários entre Pro-hemócitos e Plasmatócitos e entre Granulócitos e Adipohemócitos também foram encontrados, fazendo-se supor que pro-hemócitos dão origem ao Plasmatócito e que este dá origem ao Granulócito que pode acumular grãos de lipídeos transformando-se em Adipohemócito. O Oenocitóide parece ter origem diferente dos demais tipos. Não foram encontradas formas transicionais entre Plasmatócito fusiforme e Vermiforme típica conforme aparece na literatura para algumas espécies. Embora sem ter característica de hemócitos, as células Esfoliativas são elementos que aparecem nos dois instares estudados.

Desenvolvimento das gônadas de Dermatobia hominis (Diptera: Cuterebridae)

O trabalho descreve o desenvolvimento das gônadas do berne (D. hominis) durante o período pupal. As pupas desenvolvidas de larvas com peso superior a 650 mg, deram imagos fêmeas, enquanto que as desenvolvidas daquelas pesando entre 500 e 650 mg deram macho, tendo havido um erro ao redor de 5%. Até o oitavo dia de pupação os testículos crescem mais que os ovários; a partir daí diminui o desenvolvimento, parando de crescer entre o vigésimo e vigésimo quinto dias. A espermatogênese inicia por volta do sétimo dia de pupa quando é grande o número de espermatócitos. No décimo dia alguns testículos apresentam considerável número de espermátides e os espermatozóides começam a aparecer por volta do vigésimo dia. A espermiogênese desenvolve-se sem interrupção e ao final da pupação quase toda loja testicular está repleta de espermatózóides. Os machos começam a nascer dois dias antes das fêmeas. Nessas, os ovaríolos aparecem formados por volta do oitavo dia de pupa; os folículos se individualizam por volta do vigésimo dia de pupa onde se distingue os trofócitos com núcleos politênicos e citoplasmas bem basófilos, enquanto o ovócito tem citoplasma mais acidófilo e núcleo com cromatina bastante frouxa. A vitelogênese tem início ao redor do vigésimo quinto dia de pupa e se completa ao nascimento da imago. A ligação das gônadas com suas respectivas estruturas somáticas acontece ao redor do décimo terceiro dia de pupação.

Ultrastructure of the ovary of Dermatobia hominis (Diptera: Cuterebridae): III. Gonial cell degeneration

We studied the ultrastructural aspects of pre-pupae and pupae ovaries of Dermatobia hominis. Physiological degeneration of gonial cells was observed: (a) after the ovarioles differentiation, in the oogonia residing in the apical region of the ovary; (b) at the beginning of vitellogenesis, in the cystoblasts close to the terminal filament. The significance of gonial cell degeneration was correlated with the physiological changes wich occur in the ovary during development.

Ultrastructure of the ovary of Dermatobia hominis (Diptera: cuterebridae). I. Development during the 3rd larval instar

The ultrastructure and distribution of gonial and somatic cells in the ovary of Dermatobia hominis was studied during the 3rd larval instar. In larvae weighing between 400 and 500 mg, the ovary is partially divided into basal and apical regions by oblong somatic cells that penetrate from the periphery; these cells show ovoid nucleus and cytoplasm full of microtubules. In both regions, gonial cells with regular outlines, large nucleus and low electron-density cytoplasm are scattered among the interstitial somatic cells. These later cells have small nucleus and electrodense cytoplasm. Clear somatic cells with small nucleus and cytoplasm of very low electron-density are restrict to the apical region of the gonad. Degenerating interstitial somatic cells are seen in the basal portion close to the ovary peduncle. During all this larval period the morphological features of the ovary remain almost the same. At the end of the period there is a gradual deposition of glycogen in the cytoplasm of the somatic cells, increase in the number and density of their mitochondria plus nuclear modification as membrane wrinkling and chromatin condensation in masses.

Ultrastructure of the ovary of Dermatobia hominis (Diptera: cuterebridae). II. Origin of the tunica propria in ovarioles

Ovaries up to the 8th day pupae of Dermatobia hominis were studied by transmission electron microscopy. Ovarioles were recognized in ovaries of 4-day old pre-pupae, surrounded by a thin tunica propria of acellular fibrilar material similar in structure to the internal portion of the external tunica of the ovary. There is continuity of the tunica propria and the ovarian tunica, indicating that the former structure originates from the tunica externa. In 5 to 7-day pupae the interstitial somatic cells from the apical region of the ovary, close to the ovarioles, show delicate filamentous material inside of their rough endoplasmic reticulum cisternae; similar material is seem among these cells. Our observations suggest that interstitial somatic cells do not originate the tunica propria but contribute to its final composition.

S-100 dendritic cells in normal and Dermatobia hominis infested cattle skin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Characterization of the excretory/secretory products of Dermatobia hominis larvae, the human bot fly

Proteolytic activity in excretory/secretory products (ESP) of first- (L1), second- (L2) and third-instar (L3) larvae of Dermatobia hominis was analyzed through gelatin-gel and colorimetric enzyme assays with the chromogenic substrates azocasein and BApNA. The functional characterization of proteases was based on inhibition assays including synthetic inhibitors. ESP were obtained from new-hatched larvae reared in the laboratory and from second- and third-instar larvae removed from naturally infested cattle. Gelatin-gel analysis evidenced few bands of proteolysis, predominantly of high apparent molecular masses, in ESP of L1, whereas in the gel of L2 and U ESP there was a wide range of proteolytic activity most of them not resolved in a single species. Azocasein assays revealed a progressive increase of protease activity from first- to third-instar larvae. Protease inhibitor assays revealed a predominance of metalloproteases in L1 ESP that could be related to a skin penetration process and to a diversion of host immune response. The predominance of serine proteases in L2 and L3 and the great tryptic activity presented by L3 ESP were attributed to an increasing trophic activity by the growing larvae, since the viability of adult flies strictly depends on larval abilities to assimilate nutrients from the host. Taking together, these results suggest that Dematobia larvae secrete/excrete different proteases that may be related to diverse functions during host penetration and infestation, which reinforces the relevance of the study of such proteolytic enzymes. (C) 2009 Elsevier B.V. All rights reserved.

Occurrence of Nuages and Lamellae Anulata during spermatogenesis in Dermatobia hominis (Diptera: Cuterebridae)

Various types of nuages and lamellae anulata can be found during Dermatobia hominis spermatogenesis. In spermatogonia, the nuages occur as granules juxtaposed to the cytoplasmic face of the nuclear envelope or as cytoplasmic granules similar toglycogen granules. In spermatocytes, in addition to the nuages, dense spherical bodies of approximately 1.0 µm in diameter are also observed. In the spermatids the nuages can be of the following types: perinuclear granules, spherical granules with diameters varying in length from 0.5 to 1.0 µm, granules similar to glycogen granules, granules with variable diameters which accumulate at the flagellum base forming the centriole adjunct, or remain in the cytoplasm. Nuages can also be observed in these cellular types as dense masses, without a definite outline and are common to animal germinal cells in general. The lamellae anulata on the other hand, are observed only in spermatocytes I and in early spermatids, being always immersed in electron-dense material of indefinite outline. In spermatids, the lamellae anulata are close to the nuclear envelope suggesting, in spite of opposing opinions, that these cells are envolved in the synthesis and transport of material from the nucleus to the cytoplasm.

Post-embrionic development of the digestive tube of Dermatobia hominis (Linnaeus) (Diptera, Cuterebridae)

The digestive tube of 2nd and 3rd instar larvae, pupae and newly emerged adults of Dermatobia hominis (Linnaeus, 1781) was studied anatomically. The specimens were dissected in buffer saline under a stereomicroscope, and the digestive tubes were placed on slides and fixed in 10% buffered formalin. Each tube was measured using a micrometric eye piece, and drawings were made with camera lucida. The results showed that the midgut, the hindgut and the Malpighian tubules with their ducts grow gradually during the larval development. The oesophagus and the salivary glands with their ducts grow only during the moult from the 2nd to the 3rd instar. In the pupal period, salivary glands grow gradually but disappeared after the 20th day. After metamorphosis the digestive tube regressed. This is expected since adult D. hominis lives about nine days without feeding. This fly, similar to other calyptratae muscoid flies shows no vestige of a crop during all post-embrionic development, and the adult has no salivary glands.

Desenvolvimento pós-embrionário do intestino anterior de Dermatobia hominis (Linnaeus Jr.) (Diptera, Cuterebridae)

Foregut in D. hominis (Linnaeus Jr., 1781) as the majority of the larval Diptera somatic tissue, is made up of polytenic cells, and grows at the expenses of the polytenization of its nuclei followed by the increase in size of each cell. The oesophagus, of ectodermic origem, is interiorly covered by a chitinous squamous epithelium that rests upon a very thin basal lamina. This sheet is surrounded by thick muscle bundles. The oesophagus intussuscepts the midgut forming the cardia. The cardia, with three epithelial layers: two internal ones, of ectodermal origin and one external of endodermic origin. At the anterior portion of the cardia, between these two types of epithelium, there is a cluster of small, non polytenic cells, forming the imaginal disk of the foregut. Metamoiphosis begins at the end of the larval period with signs of nuclear degeneration of all the polytenic cells, as well as the increase in number of the imaginal disk ones. The oesophagic portion intussuscepted into the cardia, everts; its cells suffer apoptosis and are replaced by the new cells growing from the imaginal disk. The external layer cells also degenerate and are pinched off into the lumen of the very anterior portion of the midgut. The newly formed oesophagus intussuscepts de novo to form the two internal layers of the adult cardia. At the same time the midgut regenerative cells grow anteriorly to form the new external layer of the adult cardia.