The housefly Musca domestica as a mechanical vector of Clostridium difficile

Background - Clostridium difficile is a bacterial healthcare-associated infection that may be transferred by houseflies (Musca domestica) due to their close ecological association with humans and cosmopolitan nature. Aim - To determine the ability of M. domestica to transfer C. difficile both mechanically and following ingestion. Methods - M. domestica were exposed to independent suspensions of vegetative cells and spores of C. difficile, then sampled on to selective agar plates immediately postexposure and at 1-h intervals to assess the mechanical transfer of C. difficile. Fly excreta was cultured and alimentary canals were dissected to determine internalization of cells and spores. Findings - M. domestica exposed to vegetative cell suspensions and spore suspensions of C. difficile were able to transfer the bacteria mechanically for up to 4 h upon subsequent contact with surfaces. The greatest numbers of colony-forming units (CFUs) per fly were transferred immediately following exposure (mean CFUs 123.8 +/− 66.9 for vegetative cell suspension and 288.2 +/− 83.2 for spore suspension). After 1 h, this had reduced (21.2 +/− 11.4 for vegetative cell suspension and 19.9 +/− 9 for spores). Mean C. difficile CFUs isolated from the M. domestica alimentary canal was 35 +/− 6.5, and mean C. difficile CFUs per faecal spot was 1.04 +/− 0.58. C. difficile could be recovered from fly excreta for up to 96 h. Conclusion - This study describes the potential for M. domestica to contribute to environmental persistence and spread of C. difficile in hospitals, highlighting flies as realistic vectors of this micro-organism in clinical areas.

The gene transformer-2 of Anastrepha fruit flies (Diptera, Tephritidae) and its evolution in insects

Background: In the tephritids Ceratitis, Bactrocera and Anastrepha, the gene transformer provides the memory device for sex determination via its auto-regulation; only in females is functional Tra protein produced. To date, the isolation and characterisation of the gene transformer-2 in the tephritids has only been undertaken in Ceratitis, and it has been shown that its function is required for the female-specific splicing of doublesex and transformer pre-mRNA. It therefore participates in transformer auto-regulatory function. In this work, the characterisation of this gene in eleven tephritid species belonging to the less extensively analysed genus Anastrepha was undertaken in order to throw light on the evolution of transformer-2. Results: The gene transformer-2 produces a protein of 249 amino acids in both sexes, which shows the features of the SR protein family. No significant partially spliced mRNA isoform specific to the male germ line was detected, unlike in Drosophila. It is transcribed in both sexes during development and in adult life, in both the soma and germ line. The injection of Anastrepha transformer-2 dsRNA into Anastrepha embryos caused a change in the splicing pattern of the endogenous transformer and doublesex pre-mRNA of XX females from the female to the male mode. Consequently, these XX females were transformed into pseudomales. The comparison of the eleven Anastrepha Transformer-2 proteins among themselves, and with the Transformer-2 proteins of other insects, suggests the existence of negative selection acting at the protein level to maintain Transformer-2 structural features. Conclusions: These results indicate that transformer-2 is required for sex determination in Anastrepha through its participation in the female-specific splicing of transformer and doublesex pre-mRNAs. It is therefore needed for the auto-regulation of the gene transformer. Thus, the transformer/transfomer-2 > doublesex elements at the bottom of the cascade, and their relationships, probably represent the ancestral state ( which still exists in the Tephritidae, Calliphoridae and Muscidae lineages) of the extant cascade found in the Drosophilidae lineage ( in which tra is just another component of the sex determination gene cascade regulated by Sex-lethal). In the phylogenetic lineage that gave rise to the drosophilids, evolution co-opted for Sex-lethal, modified it, and converted it into the key gene controlling sex determination.

Transformation of external sensilla to chordotonal sensilla in the cut mutant of Drosophila assessed by single-cell marking in the embryo and larva

The cut gene of Drosophila melanogaster is an identity selector gene that establishes the program of development and differentiation of external sense organs. Mutations in the cut gene cause a transformation of the external sense organs into chordotonal organs, originally assessed by the use of immunostaining methods [Bodmer et al. (1987): Cell, 51:293-307]. Because of evidence that axonal projections of the transformed neurons within the central nervous system are not completely switched in cut mutants, the transformation of the four cells making up a sense organ was reassessed using single-cell staining with fluorescent dye and differential interface contrast (DIC) microscopy of the embryo and larva. The results provide strong evidence that all cells of the sense organs are completely transformed, exhibiting the morphologies and organelles characteristic of chordotonal sense organs. A comparison of the structures of external sense organs and chordotonal organs indicates that a number of the differences could be due to the degree of development of common structures, and that cut or downstream genes modulate effector genes that are normally utilized in both receptor types. The possible derivation of insect chordotonal and external sense organs from a receptor type found in crustaceans is discussed in the light of arthropod phylogenetics and the molecular genetics of sense organ development. (C) 1997 Wiley-Liss, Inc.

maT - A clade of transposons intermediate between mariner and Tc1

A group of transposons, named maT, with characteristics intermediate between mariner and Tc1 transposons, is described. Two defective genomic copies of MdmaT from the housefly Musca domestica, with 85% identity, were found flanking and imbedded in the MdalphaE7 esterase gene involved in organophosphate insecticide resistance. Two cDNA clones, with 99% identity to each other and 72%-89% identity to the genomic copies were also obtained, but both represented truncated versions of the putative open reading frame. A third incomplete genomic copy of MdmaT was also identified upstream of the putative M. domestica period gene. The MdmaT sequences showed high identity to the transposable element Bmmar1 from the silk-worm moth, Bombyx mori, and to previously unidentified sequences in the genome of Caenorhabditis elegans. A total of 16 copies of full-length maT sequences were identified in the C elegans genome, representing three variants of the transposon, with 34%-100% identity amongst them. Twelve of the copies, named CemaT1, were virtually identical, with eight of them encoding a putative full length, intact transposase. Secondary structure predictions and phylogenetic analyses confirm that maT elements belong to the mariner-Tc1 superfamily of transposons, but their intermediate sequence and predicted structural characteristics suggest that they belong to a unique clade, distinct from either mariner-like or Tc1-like elements.

A honeybee storage protein gene, hex 70a, expressed in developing gonads and nutritionally regulated in adult fat body

In preparing for metamorphosis, insect larvae store a huge amount of proteins in hemolymph, mainly hexamerins. Out of the four hexamerins present in the honeybee larvae, one, HEX 70a, exhibited a distinct developmental pattern, especially since it is also present in adults. Here, we report sequence data and experimental evidence suggesting alternative functions for HEX 70a, besides its well-known role as an amino acid resource during metamorphosis. The hex 70a gene consists of 6 exons and encodes a 684 amino acid chain containing the conserved hemocyanin N, M, and C domains. HEX 70a classifies as an arylphorin since it contains more than 15% of aromatic amino acids. In the fat body of adult workers, hex 70a expression turned out to be a nutrient-limited process. However, the fat body is not the only site for hex 70a expression. Both, transcript and protein subunits were also detected in developing gonads from workers, queens and drones, suggesting a role in ovary differentiation and testes maturation and functioning. In its putative reproductive role, HEX 70a however differs from the yolk protein, vitellogenin, since it was not detected in eggs or embryos. (C) 2008 Elsevier Ltd. All rights reserved.

Dípteros muscóides como vetores mecânicos de ovos de helmintos em jardim zoológico, Brasil

OBJETIVO: Verificar as espécies de dípteros muscóides capazes de veicular ovos e larvas de helmintos e avaliar o potencial de contaminação dos dípteros capturados. MÉTODOS: A pesquisa foi realizada em dois pontos distintos do Jardim Zoológico da cidade do Rio de Janeiro, RJ, no período de maio de 1996 a abril de 1998. As capturas dos dípteros foram realizadas semanalmente com armadilhas contendo peixe em putrefação, que permaneceram expostas durante uma hora nos dois pontos: local 1- próximo à lixeira do zoológico e o local 2- perto do recinto do hipopótamo e das aves de rapina. Foram capturadas 41.080 moscas, sendo a espécie Chrysomya megacephala mais representativa com 69,34%, seguida de Chrysomya albiceps 11,22%, Musca domestica 7,15%, Chrysomya putoria 4,52%, Fannia sp. 3,12%, Ophyra sp. 2,53% e Atherigona orientalis 2,08%. As moscas capturadas tiveram a superfície dos corpos lavadas com água destilada e os tubos digestivos dissecados. RESULTADOS: Das espécies estudadas, C. megacephala e M. domestica apresentaram maior quantidade de ovos de helmintos na superfície do corpo e no conteúdo intestinal. Ovos de Ascaridoidea e Trichinelloidea prevaleceram no conteúdo intestinal de C. megacephala. Dos ovos de helmintos encontrados na superfície do corpo e no conteúdo intestinal foram identificados: Ascaris sp., Toxascaris sp., Toxocara sp., Trichuris sp., Capillaria sp., Oxiurídeos, Triconstrogilídeos e Acantocephala. Também foram encontradas larvas de helmintos na superfície do corpo dos dípteros. Houve diferenças significativas (nível de 5%, pelo teste F) entre os diferentes pontos de capturas em relação ao número de ovos de helmintos encontrados nos dípteros. CONCLUSÕES: As fezes dos animais do jardim zoológico, encontradas freqüentemente nos abrigos e lixeiras, contribuíram para a proliferação dos dípteros muscóides, que assumem importante papel na veiculação de ovos de helmintos, principalmente pelo contato direto do corpo do díptero com o alimento dos animais.

Diptera of sanitary importance associated with composting of biosolids in Argentina

OBJECTIVE: Odorous compounds produced at the biosolids composting plant in Bariloche (NW Patagonia) attract a variety of insects, mainly belonging to the order Diptera. In order to characterize these flies, collected specimens were taxonomically identified, their community characteristics were described and their sanitary and synanthropic importance and autochthonous or introduced character were determined. METHODS: Sampling was performed from October 1999 until March 2000. Adults were collected using an entomological net, and larvae and puparia were obtained from the composting material and incubated to obtain adults. Richness, abundance and sex ratio were calculated. RESULTS: A total of 9 taxa of Diptera were identified: Sarconesia chlorogaster, Phaenicia sericata, Calliphora vicina, Cochliomya macellaria, Ophyra sp, Muscina stabulans, Musca domestica, Sarcophaga sp and Fannia sp. Specimens of Anthomyiidae, Acaliptratae and one larva of Eristalis tenax were also found. Ophyra sp. was the most abundant taxa. All the captured Diptera belonged to introduced taxa. Most of them are considered to be eusynanthropic and/or hemisynanthropic and have sanitary importance as they may cause myiasis and pseudomyiasis. The high number of females registered and the finding of immature stages indicated that flies can develop their complete life cycle on biosolid composting windrows. CONCLUSIONS: The characterization of flies obtained in this study may be useful for defining locations of urban or semi-urban composting facilities. It also highlights the importance of sanitary precautions at such plants.

Effects of eucalyptol on house fly (Diptera: Muscidae) and blow fly (Diptera: Calliphoridae)

The effects of eucalyptol were evaluated against the house fly, Musca domestica L., and blow fly, Chrysomya megacephala (F.). The bioassay of adults, using topical application, indicated that M. domestica males were more susceptible than females, with the LD50 being 118 and 177 µg/fly, respectively. A higher LD50 of C. megacephala was obtained; 197 µg/fly for males and 221 µg/fly for females. Living flies of both species yielded a shorter life span after being treated with eucalyptol. The bioassay of larvae, using the dipping method on the third instar, showed that M. domestica was more susceptible than C. megacephala, with their LC50 being 101 and 642 µg/µl, respectively. The emergence of adults, which had been treated with eucalyptol in larvae, decreased only in M. domestica. Having the volatile property, fumigation or impregnated paper test of eucalyptol or the efficacy of repellence or attractiveness merits further investigations to enhance bio-insecticidal efficacy.

Some ultrastructural superficial changes in house fly (Diptera: Muscidae) and blow fly (Diptera: Calliphoridae) larvae induced by eucalyptol oil

The ultrastructural superficial changes in third instar house fly (Musca domestica) and blow fly (Chrysomya megacephala) induced by eucalyptol oil were observed using scanning electron microscopy. Dipped in 0.902 g/ml eucalyptol for 30 sec, the larvae integument of both species showed significant aberrant appearance of the body surface, particularly swelling integument, bleb formation, partial breach and deformation of spines.

Plantas nativas como fuente de biopesticidas: explorando distintas interacciones planta-organismo blanco dirigidas al control de plagas, malezas y microorganismos.

Continuando con nuestra búsqueda de biopesticidas de potencia relevante en plantas nativas, en este proyecto nos concentraremos en encontrar sustancias que controlen eficientemente hormigas cortadoras, moscas, cucarachas, mosquitos y malezas. En la primera parte del presente proyecto se determinará mediante aislamiento bioguiado, los principios activos de las plantas que en un trabajo anterior, mostraron mayor inhibición del forrajeo de la hormiga Acromyrmex lundi, Aristolochia argentina y Lantana grisebachii, como también de aquellas que inhibieron al hongo simbionte aislado del nido, A. argentina y Flourensia oolepis. Posteriromente se evaluará la efectividad de estos extractos y principios activos a campo contra A. lundi determinando dosis efectivas para inhibir el forrajeo y/o el desarrollo del hongo simbionte. Luego se extenderá los resultados a otras especies de hormigas cortadoras como A. crassispinus y A. striatus. En segundo lugar proponemos el desarrollo de una metodología para aumentar la efectividad de aceites esenciales (AE) y terpenos (T) contra distintas plagas domésticas aprovechando la interacción de componentes de aceites esenciales en la intoxicación del insecto. Hemos demostrado que Musca domestica al ser fumigada con T puros, por separado, oxida a la mayoría de ellos mediante citocromo P450. Sin embargo cuando es fumigada con AE (mezcla de T) detoxifica al T mayoritario preferentemente, mientras que los T minoritarios están en condiciones de intoxicar al insecto ya que no serían blanco del P450. En este proyecto determinaremos la DL50 de 10 T en moscas tratadas y sin tratar con un inhibidor de P450 (butóxido de piperonilo). Con estos datos podremos elegir mezclas de terpenos tales que uno de ellos sea blanco de P450 y otro/s intoxiquen al insecto. Nos proponemos verificar este mecanismo en M. domestica y luego extenderlo a otros insectos domésticos (cucarachas, mosquitos). Este estudio facilitará el desarrollo de bioinsecticidas mas eficientes para cada insecto disminuyendo sus costos y favoreciendo su aplicación futura. En la tercera parte de este proyecto proponemos aislar los principios activos fitotóxicos de Cortadeira rudiuscula y Ophryosporus charua, cuyos extractos inhibieron selectivamente la germinación de mono- y dicotiledóneas, respectivamente. También se determinará el tiempo de vida media en suelo, y la dosis efectiva a campo con el fin de evaluar a dichas especies como futuros herbicidas naturales selectivos.

Flutuação de Diptera em granja avícola, Pelotas, Rio Grande do Sul, Brasil

In order to investigate the population fluctuation of Diptera in a poultry house in Pelotas, Rio Grande do Sul, Brazil, six collection methods were utilized: 1 (0 to 7 day-old feces from chickens), 2 (7 to 14 day-old feces), 3 (14 to 21 day-old feces), 4 (0 to 21 day-old feces), 5 (accumulated feces) and 6 (tube trap). Analyses of polynomial regression were accomplished independent of the collection method. The survey was conducted from August 1998 to July 1999 in chicken houses at the Conjunto Agrotécnico Visconde da Graça. A total of 28,720 Diptera were collected, including the following species: Coproica sp. and Telomerina flavipes (Meigen, 1830) (15,640); Drosophila repleta Wollaston, 1858 (9,229); Dohrniphora cornuta (Bigot, 1857) (2,539); Ischiolepta scabricula (Haliday, 1833) (544); Lestodiplosis sp. (320); Muscina stabulans (Fallen, 1817) (159); Musca domestica L., 1758 (143); Drosophila melanogaster Meigen, 1830 (95); Telmatoscopus albipunctatus Williston, 1893 (21); Rhegmoclema sp. (14); Fannia canicularis (L., 1761) (7); Stomoxys calcitrans (L., 1758) (2); and unidentified species of Psychodidae (6) and Muscidae (1). The greatest number of species occurred in October, November and December and the fewest in August, September and April. The greatest abundance of Diptera was recorded in October (9,092), while the lowest index of capture was noted in April (658). The population fluctuation was estimated for Coproica sp. and T. flavipes, D. repleta, D. cornuta, I. scabricula and Lestodiplosis sp.

Contribuição ao conhecimento das larvas dos Sarcophagidae com especial referência ao esqueleto cefálico (Diptera)

The author publishes a comparative study of eleven species of Sarcophagid flies and gives a redescription of the larval stages of Musca domestica L. as a model. The work was made upon material obtained from Sarcophagidae reared in the Laboratory. Some of them were parasitic flies from Insects and other invertebrates. Protodexia was reared using the domestic cockroach (Periplaneta americana) instead of the Orthroptera or Mantodea its true hosts. The larvae obtained by dissection of female abdomen was reared in flesh or agarhorse serum. The last proceeding is very good since the skins of all larval stages can be conserved for study and it is possible to observe the ecdyses. Some of the larvae prefer dead snails (Bulimulus and Fruticicola) and is able to destroy larvae of other species found in the same molluscs. The first stage maggot can be obtained by dissections of dried female specimens and furnishes very good characters to determine the species and establish the philogenetical relationship of the genera in the family. Th pseudocephalon presents very curious ornaments or grooves in some species (Oxysarcodexia). Sometimes there is a pigmented capsule covering a great part of the pseudocephalon (Titanogrypa). The cephaloskeletal sclerietes have a peculiar shape and constitution for every species, mainly in the first stage maggot.

Substratos utilizados para a criação de dípteros caliptratos em uma área urbana do Município do Rio de Janeiro

Em uma área urbana do município do Rio de Janeiro, foram criados 10.988 dípteros caliptratos, pertencentes a quatro famílias e 22 espécies. As criações foram efetuadas em quatro etapas durante os anos de 1986 a 1987. Foram utilizados como substratos: peixe, fígado, carcaça de camundongo, fezes humanas e caninas, camarão, siri, banana, manga mamaão e tomate. As espécies criadas com maior freqüência foram: Fania sp (subgrupo pusio), Atherigona orientalis, Phaenicia eximia, Paraphrissopoda chrisostoma, Chrisomyia megacephala, Ophyra solitaria, Musca domestica, Synthesiomyia nudiseta, Phaenicia cuprina, Ophyra aenescens, Sarcophagula occidua, Morellia flavicornis, e Sarcodexia innota.

Levantamento de microhimenópteros parasitóides de dípteros de importância, médico-veterinária no Brasil

De larvas e pupas de Musca domestica, Chrysomya albiceps, Cochliomyia homivorax, Stomoxys calcitrans e Syntesiomyia nudiseta coletadas em diversos ambiente, em São Paulo, Paraná, Mato Grosso, Mato Grosso do Sul e Minas Gerais, foram obtidas dez espécies de microhimenópteros parasitóides da supermamília Chalcidoidea, algumas assinaladas pela primeira vez no Brasil.

Calyptrate diptera (Muscidae and Anthomyidae) of the state of Rio de Janeiro: I. Synanthropy

The synanthropy of Muscidae and Anthomyidae was studied in three ecologically distinct areas of the metropolitan region of Rio de Janeiro. Using baits such row fish, decomposing bovine liver, fresh human faeces and mashed banana it was found that Synthesiomyia nudiseta, Atherigona orientalis, and Musca domestica are highly synanthropic in contrast with Neomuscina pictipenis, Phaonantho devia and Morellia maculipenis found exclusively in the forest.