Preparation of transgenic Musca domestica by microinjection method / 生物工程学报

The transposon vector containing enhanced green fluorescent protein (EGFP) was injected into early housefly (Musca domestica L.) eggs by microinjection method to realize stable gene expression in vivo for verification, and to study housefly gene function. A borosilicate glass micro injection needle suitable for microinjection of housefly eggs was made, the softening treatment conditions of housefly egg shells were explored, and a microinjection technology platform suitable for housefly was constructed with a high-precision microsyringe Nanoject Ⅲ as the main body. The recombinant plasmid PiggyBac-[3×P3]-EGFP containing the eye-specific 3×P3 promoter and EGFP and the stable genetic expression helper plasmid pHA3pig helper were microinjected into the treated housefly eggs. After emergence, the eye luminescence was observed, and the expression and transcription level of EGFP were detected. The results showed that the normal hatching rate of housefly eggs was 55% when rinsed in bleaching water for 35 s. The hardness of the egg shell treated for 35 s was suitable for injection and the injection needle was not easy to break. About 3% of the emerged housefly eyes had green fluorescence. Through further molecular detection, EGFP specific fragments with a size of 750 bp were amplified from DNA and RNA of housefly. Through the technical platform, the stable expression of reporter genes in housefly can be conveniently and effectively realized, and a bioreactor with housefly as the main body can be established, which provides certain reference value for subsequent research on housefly gene function.

Larvicidal and adulticidal effects and ultrastructural changes of larvae midgut epithelium of musca domestica (diptera: muscidae) fed with bacillus thuringiensis var. kyushuensis

Abstract INTRODUCTION: Musca domestica is resistant to many insecticides; hence, biological control is a suitable alternative. METHODS: We evaluated the lethality of strain Btk176 towards the larval and adult M. domestica and the histopathological effects in the larvae midgut. RESULTS: We observed 99% larval and 78.9% adult mortality within 48 hours of spore ingestion (dosage, 2.4×108 CFU/ml). The histopathological effects were consistent with cytotoxicity. PCR analysis showed the presence of the cry1Ba gene. Transmission electron microscopy revealed a bipyramidal parasporal body. Thurigiensin activity was not detected. CONCLUSIONS: The serovar, Btk176 might be a potential biocontrol agent for houseflies.

Infectivity of housefly, Musca domestica (Diptera: Muscidae) to different entomopathogenic fungi

Abstract The housefly Musca domestica is a worldwide insect pest that acts as a vector for many pathogenic diseases in both people and animals. The present study was conducted to evaluate the virulence of different local isolates of Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea on M. domestica using two bioassay techniques: (1) adult immersion and (2) a bait method applied to both larvae and adults. The results showed evidence of a broad range of responses by both stages (larvae and adults) to the tested isolates of B. bassiana, M. anisopliae and I. fumosorosea. These responses were concentration-dependent, with mortality percentages ranging from 53.00% to 96.00%. Because it resulted in lower LC50 values and a shorter lethal time, B. bassiana (Bb-01) proved to be the most virulent isolate against both housefly larvae and adults. Sublethal doses of the tested isolates were also assessed to evaluate their effect on M. domestica fecundity and longevity. The fungal infections reduced housefly survival regardless of their sex and also decreased egg production in females.

Chemical composition and insecticidal activity of Cymbopogon citratus essential oil from Cuba and Brazil against housefly / Composição química e atividade inseticida do óleo essencial de Cymbopogon citratus de Brasil e Cuba contra mosca doméstica

Essential oil of Cymbopogon citratus collected from Brazil and Cuba was tested to a chemical characterization and then was tested on the post-embryonic development of Musca domestica. The chemical composition analysis by GC-MS of the oils from Brazil/Cuba allowed the identification of 13 and 12 major constituents respectively; nine of them common to both. In the both oils, the main components were the isomers geranial and neral, which together form the compound citral. This corresponds to a total of 97.92%/Brazil and 97.69%/Cuba of the compounds identified. The monoterpene myrcene, observed only in the sample of Cuba, presented a large relative abundance (6.52%). The essential oil of C. citratus (Brazil/Cuba) was dissolved in DMSO and tested at concentrations of 5, 10, 25, 50, 75 and 100% and citral was prepared by mixing 16.8 mg with 960 µL DMSO. Both essential oils and monoterpene citral were applied topically to newly-hatched larvae (1µL/larva). The results showed a lethal concentration (LC50) of 4.25 and 3.24% for the Brazilian and Cuban essential oils, respectively. Mortalities of larval and newly-hatched larvae to adult periods were dose-dependent for the two both oils as for monoterpene citral, reaching 90%. Both essential oils and citral caused morphological changes in adult specimens.

O óleo essencial de Cymbopogon citratus, coletado no Brasil e em Cuba, foi caracterizado quimicamente e testado no desenvolvimento pós-embrionário de Musca domestica. A análise da composição química dos óleos essenciais (Brasil/Cuba), por Cromatografia Gasosa acoplada ao espectrômetro de massa (GC-EM), permitiu a identificação de 13 e 12 componentes principais, respectivamente; nove deles comuns aos dois. Em ambos os óleos, os principais componentes foram os isômeros geranial e neral, que, juntos, formam o composto citral. Esse corresponde a um total de 97,92%/Brasil e 97,69%/Cuba dos compostos identificados. O monoterpeno mirceno, observado na amostra cubana, apresentou grande abundância relativa (6,52%). O óleo de C. citratus (Brasil/Cuba) foi dissolvido em DMSO, obtendo–se as concentrações de 5, 10, 25, 75 e 100%; e o citral (16,8 mg) foi misturando com 960mL de DMSO. Tanto o óleo essencial como o monoterpeno citral foram aplicados topicamente nas neolarvas (1µL/larva). Os resultados mostraram uma concentração letal (CL50) de 4,25% e 3,24% para o óleo essencial brasileiro e cubano, respectivamente. As mortalidades do período larval e o de neo-larva a adulto foram dose-dependentes, tanto para os óleos como para o monoterpeno citral, podendo chegar a 90%. Ambos os óleos essenciais e citral causaram alterações morfológicas nos espécimes adultos.

Molecular detection and antimicrobial resistance of Pseudomonas aeruginosa from houseflies (Musca domestica) in Iran

Background Pseudomonas aeruginosa is a common bacterium that can cause disease in humans and other animals. This study was conducted to screen for molecular detection and antimicrobial-resistant P. aeruginosa in Musca domestica in different locations in the Iranian provinces of Shahrekord and Isfahan. Methods Musca domestica were captured by both manual and sticky trap methods, during the daytime, from household kitchens, cattle farms, animal hospitals, human hospitals, slaughterhouses and chicken farms at random locations in Shahrekord and Isfahan provinces of Iran, and subsequently transported to the laboratory for detection of P. aeruginosa. In the laboratory, flies were identified and killed by refrigeration in a cold chamber at −20 °C, then placed in 5 mL peptone water and left at room temperature for five hours before being processed. Pseudomonas isolates were preliminarily identified to genus level based on colony morphology and gram staining, and their identity was further confirmed by polymerase chain reaction. Results Overall blaTEM gene was recovered from 8.8 % (53/600) of the P. aeruginosa isolated from houseflies collected from the two provinces. A slightly higher prevalence (10.7 %; 32/300) was recorded in Shahrekord province than Isfahan province (7.0 %; 21/300). The locations did not differ statistically (p < 0.05) in bacterial prevalence in flies. Seasonal prevalence showed a significantly lower infection frequency during autumn. Conclusions Houseflies are important in the epidemiology of P. aeruginosa infections.

Molecular detection and antimicrobial resistance of Pseudomonas aeruginosa from houseflies (Musca domestica) in Iran

BackgroundPseudomonas aeruginosa is a common bacterium that can cause disease in humans and other animals. This study was conducted to screen for molecular detection and antimicrobial-resistant P. aeruginosa in Musca domestica in different locations in the Iranian provinces of Shahrekord and Isfahan.MethodsMusca domestica were captured by both manual and sticky trap methods, during the daytime, from household kitchens, cattle farms, animal hospitals, human hospitals, slaughterhouses and chicken farms at random locations in Shahrekord and Isfahan provinces of Iran, and subsequently transported to the laboratory for detection of P. aeruginosa. In the laboratory, flies were identified and killed by refrigeration in a cold chamber at −20 °C, then placed in 5 mL peptone water and left at room temperature for five hours before being processed. Pseudomonas isolates were preliminarily identified to genus level based on colony morphology and gram staining, and their identity was further confirmed by polymerase chain reaction.ResultsOverall blaTEM gene was recovered from 8.8 % (53/600) of the P. aeruginosa isolated from houseflies collected from the two provinces. A slightly higher prevalence (10.7 %; 32/300) was recorded in Shahrekord province than Isfahan province (7.0 %; 21/300). The locations did not differ statistically (p < 0.05) in bacterial prevalence in flies. Seasonal prevalence showed a significantly lower infection frequency during autumn.ConclusionsHouseflies are important in the epidemiology of P. aeruginosa infections.(AU)

Effectiveness of Diflubenzuron in the control of houseflies.

Background & objectives: Houseflies are one of the major pest problems worldwide. The current housefly control strategy in the country hinges on the use of Dichlorvos as a housefly larvicide and pyrethrum spray in kitchens/dining areas. The present study was undertaken to evaluate the newer housefly control options in the wake of concerns about human toxicity of Dichlorvos and its continued use in public health. Methods: Efficacy of Diflubenzuron WP and granule formulations (chitin synthesis inhibitor) visà- vis Dichlorvos EC in the control of housefly was assessed in laboratory by adult emergence inhibition after treatment and in field conditions by estimation of density post-application up to three weeks using standard methods. Results: Dichlorvos EC and Diflubenzuron formulations (WP and granule) brought about 100% inhibition of adult emergence under laboratory conditions. However, in the field evaluation, effective suppression of housefly population was brought about by Dichlorvos and Diflubenzuron granule formulation (though delayed). Conclusion: Dichlorvos EC and Diflubenzuron granule formulation are effective housefly larvicides. It is recommended that Diflubenzuron WP formulation be evaluated at a higher frequency to enhance its effectiveness and its use may be considered in conjunction with an initial suppression of housefly adults with an adulticide, e.g. Cyphenothrin, etc.

Antibacterial effect of the antibacterial peptide from housefly larvae on Enterobacter cloacae infected wound in mice / 重庆医科大学学报

Objective:To investigate the antibacterial effects of the antibacterial substances on Enterobacter cloacae infection of wound in mice.Methods:Antibacterial substances of the larvae of housefly were induced by ultrasonic treatment,and its antibacterial activity was investigated with inhibition experiment in vitro.Thirty ICR mice were enrolled in the study,and the Enterobacter cloacae infection model was re-produced by excision of the full layer of dorsal skin with anarea of 1 cm?1 cm.Then they were randomly divided into group C(control,n=10,with wet compress of isotonic saline),group M(with hydropathic compress of 100 g/L mafenide)and group A(with wet compress of 0.1 g/L antibacterial peptide).The changes of white blood cell counts in each group were determined before and 3 days after injury.Bacterial growth in muscular tissue of the wounds and the survival of the mice were observed at 3 days after injury.Results:The purified peptide was highly potent against Enterobacter cloacae in vitro.The number of leucocytes in each group was decreased after operation,with significantly less number in group C than in group M and A.Bacterial growth in muscle demonstrated a difference between treated groups and group C at 3 days [group M:(87?31)?102 CFU/g;group A:(99?32)?102 CFU/g;group C:(504?338)?103 CFU/g(]P

Purification of a New Antibacterial Protein from Housefly(Musca domestica)Larvae / 中国生物工程杂志

Insects exhibit a particular resistance to infections.The activation of the innate immune response of insects is involved in the recognition of the infectious nonself and subsequent activation of cellular and humoral reactions.In humoral reactions,insect antibacterial peptides and lysozyme are very important in resistance to infections.Housefly(Musca domestica) is one of the most important kinds of insects and it has strong ability to adapt to the adverse circumstances.It is of momentous theoretical and practical significance to research the immunity system of housefly.The methods of inducement of housefly larvae were firstly studied.Then an antibacterial protein,whose molecular weight is 28kDa,was purified from housefly larvae,induced by 30% H2O2,through saltout,Sephadex G25 column,Sephadex G75 column and CMSepharose Fast Flow column.This antibacterial peptide had activities against most of Gram positive and Gram negative bacteria.

RESEARCH OF THE SEX-SPECIFIC PHOTORECEPTORS OF THE COMPOUND EYE RELATIVE TO CHASING BEHAVIOR IN THE HOUSEFLY (MUSCA DOMESTIC A) / 解剖学报

Micro-and ultrastructure of the photoreceptors and first visual ganglion in the different regions of the compound eye of the wild male and female housefly, and the projections of the photoreceptor cells within the subsequent visual centers have been studied. The morphology of the retina and lamina ganglia in the front dorsal and the equatorial regions of the male compound eye and the front ventral region of the female one, and the projections of the retina within the lamina show the sexspecific characteristics underlying the function of chasing behavior in the housefly.

DISTRIBUTION AND PROPERTIES OF SEX-SPECIFIC PHOTORECEPTORS IN THE HOUSEFLY'S COMPOUND EYE / 解剖学报

1. The central retinular cells R_7 in the compound eye of male housefly can be divided into two types which are called here R_(7a) and R_(7b) respectively. R_(7a) approximate many features of R_(1-6) cells, which have a larger, cell body, rhabdomere and axon, terminate in the first optic neuropil (lamina). R_(7b) approximate many features of R_8 cell and project directly to the second optic neuropil (medulla). 2. The central retinular cell R_(7a) in the compound eye of male housefly distribute mainly the dorsal region of the eye, there are a few of R_(7a) in the ventral region of the eye, which only distribute in the first and second row ommatidia under equatorial line, but not found in the compound eye of female housefly.