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PubMed2005

Variability of wing size and shape in three populations of a recent Brazilian invader, Zaprionus indianus (Diptera: Drosophilidae), from different habitats.

Zaprionus indianus (Diptera: Drosophilidae) is an African species that was introduced in Brazil near the end of the 1990's decade. To evaluate the adaptive potential of morphological traits in natural populations of this recently introduced species, we have investigated wing size and shape variation at Rio de Janeiro populations only two years after the first record of Z. indianus in Brazil. Significant genetic differences among populations from three distinct ecological habitats were detected. The heritability and evolvability estimates show that, even with the population bottleneck that should have occurred during the invasion event, an appreciable amount of additive genetic variation for wing size and shape was retained. Our results also indicated a greater influence of environmental variation on wing size than on wing shape. The importance of quantitative genetic variability and plasticity in the successful establishment and dispersal of Z. indianus in the Brazilian territory is then discussed.

Animals↗
PubMed1989

Cell lines from Culicoides variipennis (Diptera: Ceratopogonidae) support replication of bluetongue virus.

Cell lines have been developed from 2-day-old embryos of the biting midge, Culicoides variipennis (Diptera: Ceratopogonidae). In North America C. variipennis is the primary insect vector of bluetongue virus (BTV), an orbivirus that causes disease of ruminants. The C. variipennis (CuVa) cells, grown in Schneider's Drosophila medium, consist primarily of a fibroblast-like cell type. CuVa cells are very hardy. They can grow over a wide range of temperature and pH and adapt to growth in minimal essential medium. BTV replicates to high titer (7.5-8.0 log10 50% tissue culture infectious doses/ml) in CuVa cells over a wide range of temperatures (19 degrees to 37 degrees C) without inducing any significant cytopathic effects. The highest BTV titers were obtained in CuVa cells grown at 25 degrees and 32 degrees C. Cells from C. variipennis can be useful for many diverse investigations.

Animals↗
PubMed1990

Fine structure and sporogonic development of a Vavraia sp. (Microsporida: Pleistophoridae) in the biting midge Culicoides edeni (Diptera: Ceratopogonidae).

A microsporidium with ultrastructural characteristics of the genus Vavraia was found in the fat body of an adult specimen of Culicoides edeni (Diptera: Ceratopogonidae) collected in northern Florida. The sporogonial stages developed within sporophorous vesicles, which contained variable numbers of oval spores at maturity. The wall of the sporophorous vesicle was composed of two electron-dense outer layers and an electron-lucent intermediate layer. Sporonts contained haplokaryotic nuclei and divided by rosette formation. Mature spores had anisofilar polar filaments and measured 3.8 +/- 0.28 microns in length and 2.2 +/- 0.16 microns in width in thick sections of resin-embedded material. This is the first report of a Vavraia sp. from a species of Culicodes.

Animals↗
PubMed1991

Parasitism of newly-hatched Aedes sierrensis (Diptera: Culicidae) larvae by Lambornella clarki (Ciliophora: Tetrahymenidae) following habitat flooding.

Host-parasite interactions between Lambornella clarki (Ciliophora: Tetrahymenidae) and its natural host, Aedes sierrensis (Diptera: Culicidae), were studied in newly flooded treeholes in northern California between 1986 and 1989. First instar host larvae hatched within 1 to 4 hr of flooding, while free-living trophonts of L. clarki appeared between 7 and 24 hr. As early as 24 hr after flooding, ciliates initiated the first parasite cycle by forming cuticular cysts on first instar larvae; by 64 hr, cysts were observed on larvae collected from all positive holes during all years. While larvae with as many as 12 cysts were observed, most supported only 1 cyst, and successful infections were established by the entry of a single ciliate into the host's hemocoel. Among treehole populations, the proportion of larvae with L. clarki cysts ranged from 2 to 100% at 48 hr indicating that enzootics and epizootics develop rapidly in newly flooded treeholes. Average attack rates from all holes by year ranged between 17.0 and 44.4%. Ciliates began entering hosts 48 to 72 hr after flooding, but some larvae escaped parasitization by molting to the second instar before ciliates penetrated the cuticle. In some treeholes, opportunistic microorganisms entered larvae with the invading ciliates and killed both the host and parasite.

Aedes↗
PubMed1987

Leishmaniasis in Brazil. XXIV. Natural flagellate infections of sandflies (Diptera: Psychodidae) in Pará State, with particular reference to the rôle of Psychodopygus wellcomei as the vector of Leishmania braziliensis braziliensis in the Serra dos Carajás.

Between July 1983 and December 1984 natural flagellate infections were found in 114 (1%) of 11,586 female phlebotomine sandflies (Diptera: Psychodidae) of 21 species. A further 1084 females of 17 other species were not infected. Identification of the organisms on a number of occasions confirms the exclusive parasite/vector relationship of Leishmania mexicana amazonensis/Lutzomyia flaviscutellata and Le. braziliensis braziliensis/Psychodopygus "wellcomei". Undescribed or unidentified Leishmania spp. were isolated from Lu. shawi, Lu. ubiquitalis, Lu. whitmani, Ps. hirsutus, Ps. paraensis Ps. "wellcomei", and trypanosomes from Lu. nordestina and Lu. trinidadensis. Flagellate infections were recorded in 8 of 21 species examined for the first time, and some were isolated directly from insects into cultures. Le. b. braziliensis was transmitted to a hamster by the bite of a wild-caught, naturally infected Ps. "wellcomei". 7 of the 35 infected Ps. "wellcomei" were allowed to oviposit and the eggs were reared to adults. Four produced Ps. wellcomei males only, confirming the rôle of this species as the major vector of Le. b. braziliensis.

Animals↗
PubMed2004

The development of Plasmodium falciparum in experimentally infected Anopheles gambiae (Diptera: Culicidae) under ambient microhabitat temperature in western Kenya.

The effect of microhabitat temperature variation on the early development of Plasmodium falciparum in experimentally infected Anopheles gambiae s.s. (Diptera: Culicidae) was studied. Batches of mosquitoes were fed artificially on gametocyteamic blood obtained from human volunteers and then held in five environmental conditions described as: (1) incubator maintained at constant temperature of 28 +/- 1 degrees C as control; (2) temperature unregulated laboratory environment; (3) screen house; (4) grass thatched mud house and (5) corrugated iron roofed mud house. Both the grass and iron roofed mud houses were real houses found in the village communities around the ICIPE Research Centre in Mbita Point, Suba District south-western Kenya. The temperature and relative humidity of these holding environments were recorded over the study period. Mosquitoes were dissected after 24 h and 7 days to enumerate ookinetes and oocysts stages, respectively in their midguts. The mean temperature observed in the temperature-unregulated laboratory (28 degrees C) was significantly higher than the temperature of the screen house (24 degrees C) while the mean temperature observed in the iron roof mud house (27 degrees C) was comparable with that in the grass-thatched mud house (27 degrees C) although the iron roof house experienced more variation (coefficient of variation, C.V., = 9.6%) and higher peaking temperatures than the grass-thatch house. The mean relative humidity for the laboratory and screen house were 23% and 32.5%, respectively, much lower than relative humidity in the incubator (73%). Relative humidity of the grass thatch hut (42%) and Iron roof hut (51%) were also lower than those of the incubator. The ookinete intensities for mosquitoes in the screen house (10.11 +/- 1.79 ookinetes/midgut) were not statistically different (P = 0.41) from those held in the laboratory (7.50 +/- 1.19 ookinetes/midgut) or in the incubator (9.89 +/- 1.47 ookinetes/midgut). The holding environments influenced the oocyst infection rates (P = 0.04) that increased from 8.4% in the screen house to 10.2% in the laboratory. The highest infection rate (12.5%) was observed in mosquitoes held in the incubator. However, the mean oocyst intensities in mosquitoes did not differ under these environments (P = 0.58). In the 'real village house' environments, the mean ookinete intensities were not statistically different between groups of mosquitoes compared to the incubator (P = 0.86). The oocyst infection rates observed in the highly fluctuating iron roof house were 9.4% as compared to 9.0% and 6.9% in the more stable and constant habitats of grass thatch house and incubator, respectively. Results show that the natural microhabitats did not influence the infections rates in mosquitoes (P = 0.62). These findings indicate that the variation in temperatures prevailing in western Kenya particularly inside the village houses do not impede the development of malaria parasites in A. gambiae mosquitoes.

Animals↗
PubMed2005

Seasonal fluctuations of phlebotomine sand fly populations (Diptera: Psychodidae) in the urban area of Marrakech, Morocco.

Phlebotomine sand flies (Diptera: Psychodidae) were collected continuously, using sticky traps, during 1 year from October 2002 to September 2003, in an urban area of Marrakech city (Morocco). A total of 3277 specimens were collected belonging to five species. Phlebotomus (Phlebotomus) papatasi (54.6%) is the predominant species followed by Sergentomyia (Sergentomyia) minuta (20%), S. (S.) fallax (11.3%), P. (Paraphlebotomus) sergenti (10.3%) and P. (Larroussius) longicuspis (3.8%). Data analyses showed a mono-modal annual pattern for P. sergenti and a bi-modal one for the other species. P. papatasi, the proven vector of Leishmania major in Morocco, was active throughout the year. This species did not diapause in this region. P. papatasi population peaked in June and November, which relating to the periods of risk in this area. Its preferred temperature ranged between 32 and 36 degrees C but no significant correlation was found between its density and the temperature. Considering the high density and long activity period of P. papatasi, the area of Marrakech should be regarded as a potential focus for L. major. This suggests the need for a continuously surveillance to prevent risk of zoonotic cutaneous leishmaniasis.

Animals↗
PubMed2005

Effects of neem limonoids on the malaria vector Anopheles stephensi Liston (Diptera: Culicidae).

The effects of the neem (Azadirachta indica A. Juss) limonoids azadirachtin, salannin, deacetylgedunin, gedunin, 17-hydroxyazadiradione and deacetylnimbin on Anopheles stephensi Liston (Diptera: Culicidae) were investigated. In exploring advantages of pure neem limonoids, we studied the larvicidal, pupicidal, adulticidal and antiovipositional activity of neem limonoids. Azadirachtin, salannin and deacetylgedunin showed high bioactivity at all doses, while the rest of the neem limonoids were less active, and were only biologically active at high doses. Azadirachtin was the most potent in all experiments and produced almost 100% larval mortality at 1 ppm concentration. In general, first to third larval instars were more susceptible to the neem limonoids. Neem products may have benefits in mosquito control programs.

Animals↗
PubMed2006

Altitudinal structuring of sand flies (Diptera: Psychodidae) in the High-Atlas mountains (Morocco) and its relation to the risk of leishmaniasis transmission.

This paper presents the results of entomological surveys on phlebotomine sand flies (Diptera: Psychodidae) in the Haouz of Marrakech and High-Atlas mountains (Morocco). Sand flies were captured with sticky traps from 25 stations with altitudes ranging between 400 and 1400 m. A total of 2742 specimens belonging to nine phlebotomine species was collected, Phlebotomus (Larroussius) perniciosus Newstead being the predominant species. There was a remarkable difference in the diversity of the sand fly fauna among the altitudes. Two associations of sand fly faunas were determined, the first one in lower altitude and the second one in higher altitude. The significance of the predominant species at any altitude range was discussed in terms of the risk of transmission of leishmaniasis.

Altitude↗
PubMed2006

Molecular identification and phylogeny of the Maculatus group of Anopheles mosquitoes (Diptera: Culicidae) based on nuclear and mitochondrial DNA sequences.

The Maculatus group of Anopheles mosquitoes (Diptera: Culicidae) comprises eight known species, including important malaria vectors in Southeast Asia. The sequences of the second internal transcribed spacer (ITS2) and third domain (D3) of ribosomal DNA, and cytochrome oxidase subunit II (COII) of mitochondrial DNA were obtained for five species of the group from China, as An. maculatus, An. willmori, An. pseudowillmori, An. sawadwongporni and An. dravidicus. The variation within taxon is much smaller than that between taxa. A diagnostic PCR assay for distinguishing the five members was developed based on the interspecific ITS2 variation. The phylogenetic relationships for the group were estimated on the ITS2 and D3 data. The Maculatus group appears monophyletic with An. pseudowillmori at a basal position, the Sawadwongporni subgroup and the Maculatus subgroup form sister clades. Our data concludes that An. dispar and An. greeni belong to the Maculatus subgroup, and An. willmori is not closer to either of the subgroups.

Animals↗
PubMed2006

The effects of blood feeding and exogenous supply of tryptophan on the quantities of xanthurenic acid in the salivary glands of Anopheles stephensi (Diptera: Culicidae).

Xanthurenic acid (XA), produced as a byproduct during the biosynthesis of insect eye pigment (ommochromes), is a strong inducer of Plasmodium gametogenesis at very low concentrations. In previous studies, it was shown that XA is present in Anopheles stephensi (Diptera: Culicidae) mosquito salivary glands and that during blood feeding the mosquitoes ingested their own saliva into the midgut. Considering these two facts together, it is therefore likely that XA is discharged with saliva during blood feeding and is swallowed into the midgut where it exerts its effect on Plasmodium gametocytes. However, the quantities of XA in the salivary glands and midgut are unknown. In this study, we used high performance liquid chromatography with electrochemical detection to detect and quantify XA in the salivary glands and midgut. Based on the results of this study, we found 0.28+/-0.05 ng of XA in the salivary glands of the mosquitoes, accounting for 10% of the total XA content in the mosquito whole body. The amounts of XA in the salivary glands reduced to 0.13+/-0.06 ng after mosquitoes ingested a blood meal. Approximately 0.05+/-0.01 ng of XA was detected in the midgut of nonblood fed An. stephensi mosquitoes. By adding synthetic tryptophan as a source of XA into larval rearing water (2 mM) or in sugar meals (10 mM), we evaluated whether XA levels in the mosquito (salivary glands, midgut, and whole body) were boosted and the subsequent effect on infectivity of Plasmodium berghei in the treated mosquito groups. A female specific increase in XA content was observed in the whole body and in the midgut of mosquito groups where tryptophan was added either in the larval water or sugar meals. However, XA in the salivary glands was not affected by tryptophan addition to larval water, and surprisingly it reduced when tryptophan was added to sugar meals. The P. berghei oocyst loads in the mosquito midguts were lower in mosquitoes fed tryptophan treated sugar meals than in mosquitoes reared on tryptophan treated larval water. Our results suggest that mosquito nutrition may have a significant impact on whole body and midgut XA levels in mosquitoes. We discuss the observed parasite infectivity results in relation to XA's relationship with malaria parasite development in mosquitoes.

Animals↗
PubMed2005

Efficacy of Melia azedarach L. extract on the malarial vector Anopheles stephensi Liston (Diptera: Culicidae).

Methanolic extracts of leaves and seeds from the chinaberry tree, Melia azedarach L. (Meliaceae) was tested against mature and immature mosquito vector Anopheles stephensi Liston (Diptera) under laboratory condition. The extract showed strong larvicidal, pupicidal, adulticidal, antiovipositional activity, repellency and biting deterency. The M. azedarach seed and leaf extracts were used to determine their effect on A. stephensi adults and their corresponding oviposition and consequent adult emergence in comparison with the control. The seed extracts showed high bioactivity at all doses, while the leaf extracts proved to be active, only in the higher dose. Results obtained from the laboratory experiment showed that the seed extracts suppressed the pupal and adult activity of A. stephensi even at low dose. In general, first and second instar larvae were more susceptible to both leaves and seed extracts. Clear dose-response relationships were established with the highest dose of 2% plant extract evoking 96% mortality. Entire development of A. stephensi was inhibited by M. azedarach treatment. Less expensive (less than 0.50 US dollars per 1 kg seed), naturally accruing bio-pesticide could be an alternative for chemical pesticides.

Animals↗
PubMed2005

Effects of Dysoxylum malabaricum Bedd. (Meliaceae) extract on the malarial vector Anopheles stephensi Liston (Diptera: Culicidae).

In recent years, use of environmentally friendly and biodegradable natural insecticides of plant origin have received renewed attention as agents for disease vector control. Methanol extracts of leaves from the Indian white cedar Dysoxylum malabaricum Bedd. (Meliaceae) were tested against mature and immature Anopheles stephensi Liston (Diptera) mosquitoes under laboratory conditions. The extract showed strong larvicidal, pupicidal, adulticidal, and antiovipositional activity. The maximum leaf extract concentration tested in this study was 4%, which produced pronounced effects. In general, first and second instars were more susceptible to leaf extract than older insects. Clear dose-response relationships were established, with the highest dose of 4% plant extract causing 97% mortality of first instars.

Analysis of Variance↗
PubMed2006

The use of Eucalyptus tereticornis Sm. (Myrtaceae) oil (leaf extract) as a natural larvicidal agent against the malaria vector Anopheles stephensi Liston (Diptera: Culicidae).

Secondary metabolites obtained from the indigenous plants with proven mosquito control potential can be used as an alternative to synthetic insecticides under the integrated vector control. The essential oil extract from the forest redgum, Eucalyptus tereticornis Sm. (Myrtaceae) was tested against mature and immature mosquito vector Anopheles stephensi Liston (Diptera) under laboratory condition. The extract showed strong larvicidal, pupicidal and adulticidal activity. The leaf oil extracts showed high bioactivity at high doses. Results obtained from the laboratory experiment showed that the leaf extracts suppressed the pupal and adult activity of Anopheles stephensi at higher doses. In general, first and second instar larvae were more susceptible to all treatments. Clear dose -response relationships were established with the highest dose of 160ppm plant extract evoking almost 100% mortality. The results obtained suggest that, in addition to their medicinal activities, E. tereticornis can also serve as a natural mosquitocide.

Animals↗
PubMed2006

Expression of heat shock protein and hemoglobin genes in Chironomus tentans (Diptera, chironomidae) larvae exposed to various environmental pollutants: a potential biomarker of freshwater monitoring.

To identify a sensitive biomarker of freshwater monitoring, we evaluated pollutant-induced expression of heat shock proteins (HSPs) and hemoglobins (Hbs) genes in the larvae of the aquatic midge Chironomus tentans (Diptera, Chironomidae). As pollutants, we examined nonylphenol, bisphenol-A, 17alpha-ethynyl estradiol, bis(2-ethylhexyl) phthalate, endosulfan, paraquat dichloride, chloropyriphos, fenitrothion, cadmium chloride, lead nitrate, potassium dichromate, benzo[a]pyrene and carbon tetrachloride. We also investigated larval growth as a physiological descriptor by measuring changes in the body fresh weight and dry weight after chemical exposure. The response of the HSPs gene expression by chemical exposure was rapid and sensitive to low chemical concentrations but it was not stressor specific. Interestingly, an increase in the expression of HSPs genes was observed not only in a stress inducible form (HSP70), but also in a constitutively (HSC70) expressed form. The expression of Hb genes showed chemical-specific responses: that is, alkyl phenolic compounds increased the expression of hemoglobin genes, whereas pesticides decreased the expression. As expected, molecular-level markers were more sensitive than physiological endpoints, suggesting that gene expression could be developed as an early warning biomarker in this animal. The overall results suggest that the expression of HSP and Hb genes in Chironomus could give useful information for diagnosing general health conditions in fresh water ecosystem. The expression of Hb genes, in particular, seems to be a promising biomarker, especially in view of the potential of Chironomus larvae as a biomonitoring species and of the physiological particularities of their respiratory pigments.

Animals↗
PubMed2006

Mercury concentration in black flies Simulium spp. (Diptera, Simuliidae) from soft-water streams in Ontario, Canada.

Total Hg in Simulium spp. (Diptera, Simuliidae) was measured in 17 soft-water streams in the District of Muskoka and Haliburton County (Ontario, Canada) during 2003 and 2004. Black flies contained 0.07-0.64 microg/g total Hg (dry weight). The methylmercury concentration was measured in 6 samples of the 17, and ranged from 58% to 93% of total Hg. The concentration of total Hg is much higher than has been found in other filter feeding insects, and represents a significant potential source of Hg to fish. Mercury concentrations in Simulium spp. at different sites were strongly positively correlated with dissolved organic carbon, and the proportion of land within each catchment that was wetland. There was also a strong negative correlation with pH. By examining Hg concentration in filter feeding insects we have found a significant entry point for Hg and MeHg into the food web.

Animals↗
PubMed2005

The role of HiTI, a serine protease inhibitor from Haematobia irritans irritans (Diptera: Muscidae) in the control of fly and bacterial proteases.

Blood-sucking arthropods are vectors responsible for the transmission of several pathogens and parasites to vertebrate animals. The horn fly Haematobia irritans irritans (Diptera: Muscidae) and the tick Boophilus microplus are important hematophagous ectoparasites that cause losses in cattle production. A serine protease inhibitor from a thorax extract of the fly H. irritans irritans (HiTI) was previously isolated, characterized and cloned. In the present study we described the expression, purification, and characterization of the recombinant HiTI (rHiTI) and its possible role in the control of different endogenous and bacterial proteases. rHiTI was successfully expressed using the pPIC9 expression vector with a yield of 4.2 mg/L of active rHiTI. The recombinant HiTI purified by affinity chromatography on trypsin-Sepharose had a molecular mass of 6.53 kDa as determined by LS-ESI mass spectrometry and inhibition constants (Kis) similar to those of native HiTI for bovine trypsin and human neutrophil elastase of 0.4 and 1.0 nM, respectively. Purified rHiTI also showed inhibitory activity against the trypsin-like enzyme of H. i. irritans using its possible natural substrates, fibrinogen and hemoglobin; and also inhibited the OmpT endoprotease of Escherichia coli using fluorogenic substrates. The present results confirm that HiTI may play a role in the control of fly endogenous proteases but also suggest a role in the inhibition of pathogen proteases.

Amino Acid Sequence↗
PubMed2003

Proventriculus (cardia) plays a crucial role in immunity in tsetse fly (Diptera: Glossinidiae).

Fat body and hemocytes play a central role in cellular and humoral responses for systemic infections in invertebrates, similar to the mammalian liver and blood cells. Epithelial surfaces, in particular the midgut, participate in the initial local immune responses in order to aid in the generation of the terminal cytotoxic molecules that mediate non-self recognition. Here, we describe for the first time the immune responses of a cluster of cells at the foregut/midgut junction--known as proventriculus (cardia) in the medically and agriculturally important insect, tsetse fly (Diptera: Glossinidae). We provide evidence for the transcriptional induction of the antimicrobial peptides attacin and defensin as well as for the reactive nitrogen intermediate (RNI) nitric oxide synthase (NOS) upon microbial challenge by either microinjection or feeding. Proventriculus from immune challenged flies also has higher NOS and nitric oxide (NO) activities as well as increased levels of the reactive oxygen intermediate (ROI), hydrogen peroxide (H2O2). In several vector pathogen systems, including tsetse flies and African trypanosomes, stimulation of systemic responses prior to pathogen acquisition has been shown to reduce disease transmission. Furthermore, the induction of systemic immune responses has been documented while pathogens are still differentiating within the midgut environment. While evidence for a close molecular communication between the local and systemic responses is accumulating, the molecular signals that mediate these interactions are at present unknown. Reactive intermediates such as NO or H2O2 may function as immunological signals for mediating the molecular communication between the different insect compartments. We discuss the putative role of the proventriculus in invertebrate immunity and specifically speculate on its significance for trypanosome transmission in tsetse.

Animals↗