Gravid Flies Research Articles

The ongoing risk of Leishmania donovani transmission in eastern Nepal: an entomological investigation during the elimination era

BackgroundVisceral leishmaniasis (VL), a life-threatening neglected tropical disease, is targeted for elimination from Nepal by the year 2026. The national VL elimination program is still confronted with many challenges including the increasingly widespread distribution of the disease over the country, local resurgence and the questionable efficacy of the key vector control activities. In this study, we assessed the status and risk of Leishmania donovani transmission based on entomological indicators including seasonality, natural Leishmania infection rate and feeding behavior of vector sand flies, Phlebotomus argentipes, in three districts that had received disease control interventions in the past several years in the context of the disease elimination effort.MethodsWe selected two epidemiologically contrasting settings in each survey district, one village with and one without reported VL cases in recent years. Adult sand flies were collected using CDC light traps and mouth aspirators in each village for 12 consecutive months from July 2017 to June 2018. Leishmania infection was assessed in gravid sand flies targeting the small-subunit ribosomal RNA gene of the parasite (SSU-rRNA) and further sequenced for species identification. A segment (~ 350 bp) of the vertebrate cytochrome b (cytb) gene was amplified from blood-fed P. argentipes from dwellings shared by both humans and cattle and sequenced to identify the preferred host.ResultsVector abundance varied among districts and village types and peaks were observed in June, July and September to November. The estimated Leishmania infection rate in vector sand flies was 2.2% (1.1%–3.7% at 95% credible interval) and 0.6% (0.2%–1.3% at 95% credible interval) in VL and non-VL villages respectively. The common source of blood meal was humans in both VL (52.7%) and non-VL (74.2%) villages followed by cattle.ConclusionsOur findings highlight the risk of ongoing L. donovani transmission not only in villages with VL cases but also in villages not reporting the presence of the disease over the past several years within the districts having disease elimination efforts, emphasize the remaining threats of VL re-emergence and inform the national program for critical evaluation of disease elimination strategies in Nepal.Graphical 

Laboratory breeding of two Phortica species (Diptera: Drosophilidae), vectors of the zoonotic eyeworm Thelazia callipaeda

BackgroundSome species of drosophilid flies belonging to the genus Phortica feed on ocular secretions of mammals, acting as biological vectors of the zoonotic eyeworm Thelazia callipaeda. This study describes an effective breeding protocol of Phortica variegata and Phortica oldenbergi in insectary conditions.MethodsAlive gravid flies of P. oldenbergi, P. variegata and Phortica semivirgo were field collected in wooded areas of Lazio region (Italy) and allowed to oviposit singularly to obtain isofamilies. Flies were maintained in ovipots (200 ml) with a plaster-covered bottom to maintain high humidity level inside. Adult feeding was guaranteed by fresh apples and a liquid dietary supplement containing sodium chloride and mucin proteins, while larval development was obtained by Drosophila-like agar feeding medium. The breeding performances of two media were compared: a standard one based on cornmeal flour and an enriched medium based on chestnut flour. All conditions were kept in a climatic chamber with a photoperiod of 14:10 h light:dark, 26 ± 2 °C and 80 ± 10% RH.ResultsFrom a total of 130 field-collected Phortica spp., three generations (i.e. F1 = 783, F2 = 109, F3 = 6) were obtained. Phortica oldenbergi was the species with highest breeding performance, being the only species reaching F3. Chestnut-based feeding medium allowed higher adult production and survival probability in both P. oldenbergi and P. variegata. Adult production/female was promising in both species (P. oldenbergi: 13.5 F1/f; P. variegata: 4.5 F1/f).ConclusionsThis standardized breeding protocol, based on controlled climatic parameters and fly densities, together with the introduction of an enriched chestnut-based feeding medium, allowed to investigate aspects of life history traits of Phortica spp. involved in the transmission of T. callipaeda. Obtaining F3 generation of these species for the first time paved the road for the establishment of stable colonies, an essential requirement for future studies on these vectors in controlled conditions.Graphical abstract

Mutualism promotes insect fitness by fungal nutrient compensation and facilitates fungus propagation by mediating insect oviposition preference.

Penicillium and Bactrocera dorsalis (oriental fruit fly, Hendel) are major pathogens and pests of citrus fruits, as both of them can cause detrimental losses in citrus production. However, their interaction in the cohabitation of citrus fruits remains elusive. In this study, we revealed a mutualistic relationship between Penicillium and B. dorsalis. We found that insect behaviors can facilitate the entry of fungal pathogens into fruits, and fungal pathogens promote the fitness of insects in return. More specifically, Penicillium could take advantage of the openings left by ovipositors of flies, and adult flies contaminated with Penicillium could spread the fungus to new sites. Moreover, the volatile emissions from fungi could attract gravid flies to the infected site for egg laying. The fungus and B. dorsalis were able to establish mutual interaction, as revealed by the presence of Penicillium DNA in intestinal tracts of flies throughout all larval stages. The fungal partner seemed to promote the emergence rate and shorten the emergence duration of the flies by providing pyridoxine, one of the B group vitamins. Different from previously reported scenarios of strong avoidance of Drosophila and attraction of Aedes aegypti toward Penicillium, our findings unveil a hitherto new paradigm of the mutualism between Penicillium and B. dorsalis, by which both insect and fungus earn benefits to facilitate their propagation.

Chemical Ecology of Oviposition Dynamics in Drosophila suzukii (Diptera: Drosophilidae).

Massive economic damage by spotted-wing drosophila (SWD), Drosophila suzukii, results from their unique egg laying behavior wherein a gravid fly pierces a ripening fruit to lay a number of eggs. Gravid SWD flies employ a complex suite of behaviors to find a fruit with the optimum firmness and chemistry. We investigated chemosensory cues potentially contributing to the oviposition behavior of SWD. In a series of experiments, we studied how the fruit ripeness and the underlying volatile chemistry influence oviposition. We tested the comparative attraction of three common fruits implicated in oviposition and determined raspberries to be most attractive in the trap choice assays that strictly measured olfactory preference. Since SWD oviposit in ripening fruits and appear to avoid the overripe fruit, we further evaluated the effect of ripeness on gravid fly attraction. Overripe fruits were significantly more attractive compared to the ripe fruits. The trap choice assays were repeated in an olfactory T-maze paradigm that provided a complex odor environment, potentially experienced by the gravid flies, and the results were mostly comparable. Since our behavioral paradigms indicated a clear olfactory preference for specific ripeness stages (ripe and overripe), we analyzed the constituent volatile odorants from the three ripening stages, revealing discrete odor profiles. Finally, we quantified the total soluble sugars and carbon dioxide concentrations from field-collected raspberries in underripe, ripe, and overripe conditions, revealing that the overripe stage is the most sugar-rich. Together, our results indicate unique chemosensory adaptations in gravid SWD flies for successfully exploiting optimal oviposition resources.

Ridge gourd volatiles are attractive to gravid female melon fly, Zeugodacus cucurbitae (Coquillett) (Diptera: Tephritidae)

AbstractThe melon fruit fly, Zeugodacus cucurbitae (Coquillett) (Diptera: Tephritidae), is an important pest of cucurbits throughout India, South‐East Asia and Pacific islands. Attractants of female melon fly are of relevance as they could enhance the efficacy of current pest management practices. Previous studies have revealed several host fruits to be attractive to gravid female melon flies. However, studies on the chemical compounds responsible for their attraction are meagre. As an initial part of our study, we evaluated the response of gravid female melon flies to headspace volatiles of four different cucurbit fruits, namely bitter gourd, cucumber, ridge gourd and snake gourd. Electrophysiological and multiple‐choice olfactometer assays revealed that gravid melon flies significantly preferred ridge gourd volatiles over volatiles of other cucurbit fruits. Coupled gas chromatography‐electroantennogram detection (GC‐EAG) analysis of headspace volatiles of ridge gourd to gravid melon flies identified six EAG‐active compounds, namely α‐pinene, 1‐octen‐3‐ol, p‐cymene, p‐ethyl‐benzaldehyde, methyl salicylate and p‐cymen‐7‐ol. Individual EAG‐active compounds, except α‐pinene, were significantly attractive to gravid female flies. Moreover, a synthetic blend with the concentration and ratio of compounds as in the ridge gourd headspace samples was attractive to flies. Though, in a dual‐choice test, the preference of gravid melon flies to a synthetic blend was not significantly different from preference to ridge gourd volatile samples. Taken together, our results show that the synthetic blend of ridge gourd volatiles may be used to develop a potential lure for female melon fly, Z. cucurbitae.

Oviposition-Site Selection of Phlebotomus papatasi (Diptera: Psychodidae) Sand Flies: Attraction to Bacterial Isolates From an Attractive Rearing Medium

Phlebotomine sand flies are worldwide vectors of Leishmania parasites as well as other bacterial and viral pathogens. Due to the variable impact of traditional vector control practices, a more ecologically based approach is needed. The goal of this study was to isolate bacteria from the most attractive substrate to gravid Phlebotomus papatasi Scopoli sand flies and determine the role of bacterial volatiles in the oviposition attractancy of P. papatasi using behavioral assays. We hypothesized that gravid sand flies are attracted to bacterially derived semiochemical cues associated with breeding sites. Bacteria were isolated from a larvae-conditioned rearing medium, previously shown to be highly attractive to sand flies. The isolated bacteria were identified by amplifying and sequencing 16S rDNA gene fragments, and 12 distinct bacterial species were selected for two-choice olfactometer bioassays. The mix of 12 bacterial isolates elicited strong attraction at the lower concentration of 107 cells per ml and significant repellence at a high concentration of 109 cells per ml. Three individual isolates (SSI-2, SSI-9, and SSI-11) were particularly attractive at low doses. In general, we observed dose-related effects, with some bacterial isolates stimulating negative and some positive dose-response curves in sand fly attraction. Our study confirms the important role of saprophytic bacteria, gut bacteria, or both, in guiding the oviposition-site selection behavior of sand flies. Identifying the specific attractive semiochemical cues that they produce could lead to development of an attractive lure for surveillance and control of sand flies.

The egg and larval pheromone dodecanoic acid mediates density-dependent oviposition of Phlebotomus papatasi

BackgroundGravid females assess the conditions of oviposition sites to secure the growth and survival of their offspring. Conspecific-occupied sites may signal suitable oviposition sites but may also impose risk due to competition or cannibalism at high population density or heterogeneous larval stage structure, respectively. Chemicals in the habitat, including chemicals emitted from other organisms, serve as cues for females to assess habitat conditions. Here, we investigated the attraction and oviposition preference of the Old World cutaneous leishmaniasis vector, Phlebotomus papatasi, to young and old conspecific stages, including eggs and evaluated the effect of a semiochemical associated with eggs and neonate larvae.MethodsAttraction and oviposition preference of Ph. papatasi to each of various life stages (eggs, first-, second-, third-, fourth-instar larvae, pupae and male and female adults) was investigated using cage and oviposition jar behavioral assays. Identification of organic chemical compounds extracted from eggs was performed using GC-MS and chemicals were tested in the same behavioral assays in a dose-response manner. Behavioral responses were statistically analyzed using logistic models.ResultsGravid Ph. papatasi females were significantly attracted to and preferred to oviposit on medium containing young life stages (eggs and first instars). This preference decreased towards older life stages. Dose effect of eggs indicated a hump-shaped response with respect to attraction but a concave-up pattern with respect to oviposition. Chemical analysis of semiochemicals from eggs and first-instar larvae revealed the presence of dodecanoic acid (DA) and isovaleric acid. Sand flies were attracted to and laid more eggs at the lowest DA dose tested followed by a negative dose-response.ConclusionsFindings corroborated our hypothesis that gravid sand flies should prefer early colonized oviposition sites as indicators of site suitability but avoid sites containing older stages as indicators of potential competition. Findings also supported the predictions of our hump-shaped oviposition regulation (HSR) model, with attraction to conspecific eggs at low-medium densities and switching to repellence at high egg densities. This oviposition behavior is mediated by DA that was identified from surface extracts of both eggs and first-instar larvae. Isovaleric acid was also found in extracts of both stages.

Volatiles from Merino fleece evoke antennal and behavioural responses in the Australian sheep blow fly Lucilia cuprina.

To identify flystrike-related volatile compounds in wool from Merino sheep, the attractiveness of wool to Lucilia cuprina Wiedmann (Diptera: Calliphoridae) was examined. First, a selection of wool samples guided by previous knowledge of sheep lines, predicted to be more susceptible or more resistant to flystrike, was tested. The attractiveness of the 10 samples selected was not associated with field susceptibility: two samples from the more resistant line were identified as most attractive and two samples from the more susceptible line were identified as least attractive, based on the behavioural assays with gravid flies. Comparison of the headspace volatiles of these samples, using solid phase microextraction and gas chromatography-mass spectrometry-electroantennographic detection, revealed octanal and nonanal to be present in the attractive wool samples that elicited responses from the fly antenna. Furthermore, the two compounds were not present in wool that was least attractive to L. cuprina. In laboratory bioassays, octanal and nonanal evoked antennal and behavioural responses in gravid L. cuprina, thus confirming their potential role as semiochemicals responsible for attracting L. cuprina to Merino sheep.

The Effects of Diets and Long-term Laboratory Rearing on Reproduction, Behavior, and Morphology of Lucilia cuprina (Diptera: Calliphoridae).

The Australian sheep blow fly, Lucilia cuprina (Wiedemann), is commonly reared in the laboratory for many sequential generations on simple, fixed diets, so it can be used in veterinary, medical, and forensic studies. To investigate the effect of diet and long-term laboratory rearing on L. cuprina, flies were fed with two different diets (sugar and milk-sugar) over a year and F1, F6, and F11 generations were used for comparisons based on the number of eggs, attraction to wool and liver, and wing size. The results showed that the number of eggs of gravid flies, and the attractiveness of wool and liver did not differ significantly between diets and generations, but gravid flies were more attracted to wool and liver than non-gravid flies (P < 0.05). Moreover, in the F1 generation, thorax length and wing aspect ratio were significantly longer than in the F6 and F11 generations (P < 0.05), and the wing length was significantly longer than in the F11 generation (P < 0.05). It was concluded that neither diet nor long-term laboratory rearing affect potential fecundity or the behavioral responses of L. cuprina, but the gravidity of flies affects their behavioral response, and long-term laboratory rearing significantly affects fly morphology, apparently explaining a loss in flight performance.

Behavior and Electrophysiological Response of Gravid and Non-Gravid Lucilia cuprina (Diptera: Calliphoridae) to Carrion-Associated Compounds.

The Australian blow fly, Lucilia cuprina Wiedmann (Diptera: Calliphoridae), is a major cause of myiasis (flystrike) in Merino sheep in Australia and New Zealand and, as a primary colonizer of fresh carrion, also an important species in forensic investigations. Olfaction is considered the most important cue for insects to rapidly locate carrion over long distances, so the first carrion visitors are predicted to be very sensitive to carrion-related volatile compounds. We studied the responses of the Australian blow fly, Lucilia cuprina, to the carrion-associated compounds dimethyl trisulfide (DMTS), butyric acid, 1-octen-3-ol and indole. We also tested 2-mercaptoethanol, a compound commonly used in fly traps in Australia. We investigated whether responses of the flies are affected by their ovarian status by comparing responses of gravid and non-gravid L. cuprina in electroantennography (EAG) and two-choice laboratory bioassays. All four compounds evoked an EAG response, while only DMTS evoked responses in gas chromatography-mass spectrometry electroantennographic detection (GCMS-EAD) analyses and two-choice bioassays. Gravid flies detected lower doses of the test compounds than non-gravid flies. Our results indicate that DMTS is an important semiochemical for L. cuprina to locate carrion resources, and has potential for use in fly traps for flystrike control. Our observations also suggest that the greater sensitivity of gravid L. cuprina allows them to find fresh carrion quickly to maximize reproductive success by avoiding unsuitable degraded carrion.

Physical and Physiological Factors Influence Behavioral Responses of Cochliomyia macellaria (Diptera: Calliphoridae) to Synthetic Attractants.

Volatile chemicals from waste artificial larval media as well as from bovine blood inoculated with bacteria isolated from screwworm-infested wounds attract gravid females of Cochliomyia hominivorax Coquerel and Cochliomyia macellaria (F.). Chemicals identified from volatiles are dimethyl disulfide, dimethyl trisulfide, phenol, p-cresol, and indole; a blend of these attracted females to oviposit. Present studies investigated the effectiveness of these compounds, either in a blend or individually as potential oviposition attractants. Tests were conducted to determine the effects of gender, ovarian age, and the color and type of substrates on attraction response and oviposition of C. macellaria adults. Results showed that substrates treated with dimethyl trisulfide (DMTS) alone or the five-compound blend alone attracted significantly more gravid females than other chemicals. Black substrates treated with DMTS attracted more gravid flies than did the yellow substrates. Yellow substrates treated with indole attracted more males and nongravid females. In oviposition tests, females deposited significantly more eggs on meat-based substrates than those without meat. These findings suggest that several factors have to be considered for developing an effective oviposition attractant that should include effectiveness of individual chemicals used, the ratio of the chemicals in a blend, and their concentrations. Also, an effective trap design will need to consider using suitable color which will selectively attract gravid females.

Darkness as factor influencing the oviposition delay in Calliphora vicina (Diptera: Calliphoridae)

Many environmental and intrinsic factors (e.g. limited access to the body) can disrupt insect activity, causing a delay in the colonization of a corpse. These elements could hinder an accurate estimation of the minimum Post-mortem Interval (minPMI), raising questions about the limits of forensic entomology. Blow fly are considered mainly diurnal and relatively inactive at night, at extreme temperatures and in dark conditions. Data on their ability to lay eggs in darkness and in laboratory conditions are scarce. Oviposition by Calliphoridae during the day but in darkness has been documented in chimneys, cellars and cars. To investigate delays in oviposition in the dark we carried out laboratory experiments using plastic boxes containing Calliphora vicina Robineau-Desvoidy specimens placed in a climatic chambers at different temperatures. We found that C. vicina laid eggs in complete darkness inside the plastic boxes, but later than the specimens inside the boxes at light condition. We believe that oviposition can occur in dark indoor environments in conditions of optimal air temperature, gravid flies and an accessible corpse. However, when corpses are discovered in dark environments, entomologists should consider a significant delay in oviposition by blow fly in order to reduce errors in PMI estimation.

Differential expression profiles of the salivary proteins SP15 and SP44 from Phlebotomus papatasi.

BackgroundSand fly saliva has been shown to help parasite establishment and to induce immune responses in vertebrate hosts. In the current study, we investigated the pattern of expression of two Phlebotomus papatasi salivary transcripts in specific physiological and seasonal conditions at a hyperendemic area of zoonotic cutaneous leishmaniasis (ZCL) in Iran.MethodsSand flies were collected during 2012–2013, and grouped according to physiological stages such as unfed, fed, semi-gravid, gravid, parous, nulliparous, infected or non-infected with Leishmania major and also based on the season in which they were collected. Quantitative Real-Time PCR was applied for assessment of the expression of two relevant salivary transcripts, PpSP15 and PpSP44, associated to protection from and exacerbation of ZCL, respectively.ResultsThe expression of PpSP15 and PpSP44 transcripts was significantly up-regulated (1.74 and 1.4 folds, respectively) in blood fed compared to unfed flies. Among four groups of fed, unfed, semi-gravid and gravid flies, the lowest levels of PpSP15 and PpSP44 expression were observed in gravid flies. Additionally, the expression levels of both PpSP15 and PpSP44 transcripts in P. papatasi collected during summer were significantly up-regulated (3.7 and 4.4 folds, respectively) compared to spring collections. In addition, the PpSP15 transcript exhibited a significant up-regulation (P < 0.05) in non-infected flies compared to those infected with L. major.ConclusionsThis study contributes to our knowledge of the differential expression of salivary genes among different groups within a P. papatasi population under natural field conditions. Cutaneous and visceral leishmaniasis are of public health importance in many parts of Iran and neighbouring countries where P. papatasi is the proven and dominant sand fly vector for ZCL, the most prevalent and endemic form of the disease in Iran. Therefore, the current study could be helpful in understanding the influence of salivary genes on Leishmania transmission by phlebotomine sand flies. Our findings demonstrate the differential expression of salivary transcripts under various physiological conditions potentially influencing the sand fly capacity for parasite transmission as well as the outcome of disease.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1633-z) contains supplementary material, which is available to authorized users.

Attraction and oviposition preferences of Phlebotomus papatasi (Diptera: Psychodidae), vector of Old-World cutaneous leishmaniasis, to larval rearing media.

BackgroundAs part of a project aimed at developing oviposition attractants for the control and surveillance of Phlebotomus papatasi (a vector of Old-World cutaneous leishmaniasis), we tested the hypothesis that gravid sand flies are attracted to chemical cues emanating from the growth medium of conspecific larvae - predominantly larvae-conditioned host feces that represents a suitable oviposition site. We report the results of a systematic assessment of media from various developmental stages of the sand fly using oviposition and olfactometer behavioral assays.MethodsWe conducted multiple-choice oviposition assays in 500 mL Nalgene jars. Six treatments were placed on separate filter paper discs at the bottom of the jar: 2nd/3rd larval instar medium, 4th larval instar/pupae medium, frass from expired colonies, larval food (aged rabbit chow and rabbit feces mix), rabbit feces, and a solvent (water) control. Fifty gravid females were introduced into each jar. Cumulative number of eggs laid on each filter paper per jar was counted at different time intervals from digital images. Attraction of gravid sand flies to these six treatments was assayed with a 3-chamber linear olfactometer. Twenty gravid females were transferred to the middle chamber of the olfactometer and their distribution in treatment and control chambers was recorded after 3 h.ResultsAlmost no eggs were oviposited during the first 72 h following a blood-meal. Cumulative egg deposition increased drastically in the next 24 h (hours 73–96), with a slight non-significant increasing trend thereafter. Comparing mean cumulative egg deposition among the six treatments, we found that significantly more eggs were oviposited on 2nd/3rd larval rearing medium followed by 4th instar/pupae rearing medium. Oviposition preference did not vary over time. The olfactometer results were consistent with the oviposition assays, with 2nd/3rd larval rearing medium being the most attractive, followed by 4th instar/pupae rearing medium.ConclusionThe key finding of this study is that gravid, laboratory reared, Ph. papatasi sand flies are significantly more attracted to rearing medium of the most biologically active larval stages (2nd/3rd instar and 4th instar/pupae). This finding indicates that sand fly-digested host food and feces is attractive to gravid females and suggests that the larvae and larval gut microbiome may be involved in conditioning the oviposition substrate and possibly the production of oviposition attractants and stimulants.

Evaluation of the Host Status of Mature Green Papayas 'Maradol' for the Mexican Fruit Fly (Diptera: Tephritidae).

The suitability of mature green 'Maradol' papaya as a host of Anastrepha ludens (Loew) was studied under field and laboratory conditions. Field tests were conducted on commercial-ripened and spot-ripened fruit in two orchards and during two seasons in the state of Chiapas. Fruits at exportation ripeness are in "commercial ripeness", while fruits that are harvested immediately preceding exportation ripeness are in "spot ripeness." The field tests consisted of forced infestation experiments that evaluated papayas at two ripeness stages: the commercial- or exportation-ripened fruit (green fruits with one or two yellow stripes) and fruit before exportation ripeness called "spot ripeness." These tests were conducted in two orchards and during two seasons in the state of Chiapas, Mexico. Laboratory trials were performed with commercial-ripened fruit only. Fruit from four different postharvest periods (3, 24, 48, and 72 h) were exposed to groups of gravid flies. No larvae emerged from the fruit that was collected in the field experiments. However, some larvae and several fertile flies were obtained from the commercial-ripened fruit 72 h postharvest but not 3, 24, and 48 h postharvest in the laboratory. The results of this study indicate that the commercially ripe fruits of papaya Maradol were resistant to or free from infestation of A. ludens flies under field conditions, though these fruits must be considered nonnatural, conditional host because they became infested in the laboratory.

Nonhost status of mangosteen to Bactrocera dorsalis and Bactrocera carambolae (Diptera: Tephritidae) in Thailand.

Postharvest quarantine treatments (irradiation or vapor heat) are used to control fruit flies and other pests in mangosteen (Garcinia mangostana L) exported to the United States and Japan from Thailand. No-choice tests were conducted in the laboratory to determine whether Thai mangosteen is a host for Bactrocera dorsalis (Hendel) (oriental fruit fly) and Bactrocera carambolae Drew & Hancock (carambola fruit fly). Ripe commercial quality fruit (1 wk after harvest) that were either undamaged or damaged by puncturing or peeling the pericarp were exposed to a high density of gravid flies in screen cages and then held for 10 d and dissected to inspect for immature life stages. Undamaged mangosteen fruit were not infested by B. dorsalis and B. carambolae. Partially damaged fruit with shallow punctures in the pericarp that did not extend to the aril also were not infested. Both fruit flies could infest damaged fruit if the pericarp damage allowed oviposition in the aril. Results suggest that natural infestation of mangosteen by B. dorsalis and B. carambolae can only occur if fruit exhibit physical cracks or mechanical injury. Resistance appears to be due to the pericarp hardness and thickness as well as latex secretion. Nonhost status could be used without additional quarantine measures to achieve quarantine security against B. dorsalis and B. carambolae in mangosteen exported from Thailand.

Volatiles from waste larval rearing media attract gravid screwworm flies (Diptera: Calliphoridae) to oviposit.

Gravid screwworm flies, Cochliomyia hominivorax (Coquerel), are attracted to the volatiles from waste larval rearing media to deposit eggs. Studies were conducted to identify volatile chemicals from the waste larval media and determine their effectiveness to attract gravid flies to oviposit. Volatiles were collected using solid-phase microextraction method, and five active chemicals, namely, dimethyl disulfide, dimethyl trisulfide, phenol, p-cresol, and indole, were identified using gas chromatography-mass spectrometry. In electroantennography studies, antennae ofgravid screwworm flies, Cochliomyia macellaria (F.), responded positively to each of the identified compounds. A synthetic blend of these five compounds in the ratio of 335:200:57:1:12 was prepared and tested for its effectiveness to attract both C. hominivorax and C. macellaria using laboratory bioassay methods. Significantly more gravid C. macellaria were attracted to and landed on substrates treated with 10-fold diluted blends compared with those landed on substrates treated with ethanol only (as control). Only a few young females and young and old males were attracted to the substrates treated with the synthetic blend. The C. hominivorax females laid significantly more eggs on substrates treated with waste media, 10-fold diluted blend, and 100-fold diluted blend than on substrates with undiluted blend or ethanol. Similarly, C. macellaria deposited significantly more eggs on substrates treated with waste media, 10-fold diluted blend, and 100-fold diluted blend compared with substrates with undiluted blend or ethanol. C. macellaria females deposited significantly less amount of eggs than did C. hominivorax females. These results indicate that the synthetic blend of five compounds identified may serve as an oviposition attractant for C. hominivorax as well as for C. macellaria.

Gonotrophic Development and Survival in Field Populations of &lt;I&gt;Musca domestica&lt;/I&gt; (Diptera: Muscidae) at Dairies in California, Minnesota, and Georgia, and the Relationship of Fly Age to Relative Abundance of (&lt;I&gt;Z&lt;/I&gt;)-9-Tricosene (Muscalure)

Adult female Musca domestica L. were collected in 2004 and 2005 from dairies in California, Minnesota, and Georgia. Relative abundance of (Z)-9-tricosene (muscalure) among the dominant eight hydrocarbons was determined. Fly heads then were removed to quantify pterin levels and estimate fly age, abdomens were dissected to score gonotrophic development and parity (follicular relics), and spermathecae were examined for sperm. Daily survival was assessed using two estimates of time required to become gravid: laboratory-based degree-day (DD) estimates and estimates based on pterin values in field-collected flies matched to their stages of gonotrophic development. Among newly emerged females (oocyte stage 1) with detectable muscalure, it comprised < approximately 1.5% of cuticular hydrocarbons. In muscalure-positive flies, muscalure comprised a higher proportion of cuticular hydrocarbons in older flies from California and Minnesota (6-9% when gravid) versus flies from Georgia (<2% when gravid). Females mated in early-intermediate stages of egg development. Life expectancy, using laboratory-derived estimates of time needed to become gravid, ranged from 3.6 to 10.6 d. Using equivalent pterin-based time estimates, life expectancy ranged from 4.0 to 19.5 d. Mean DD ages (12 degrees C threshold) of gravid flies varied widely (53-95 DD) and were congruent with laboratory-based estimates (52-57 DD) in only 7 of 12 farm-year combinations. Thus, house flies under natural conditions often required more time to develop eggs than laboratory models would predict, extending daily survival estimates based on gonotrophic age by 11-74%.

Black fly attractants and the control of Onchocerciasis

Onchocerciasis or river blindness disease is a parasitic disease caused by infection from the nematode Onchocerca volvulus. The parasite is transmitted to humans by black fly vectors of the genus Simulium. Most of the infections occur in central Africa, with significant incidence also in Central and South America. The World Health Organization (WHO) estimates 18 million people suffer from onchocerciasis. However, due to a prolonged period from infection to hosts becoming symptomatic it is difficult to accurately access infection. The current method for monitoring the spread employs human bait, which is neither optimal nor ethically sound. The need for a new monitoring method is very important. It was noticed that anthropophilic gravid flies are attracted to both egg masses recently deposited by other flies of the same species as well as human scent. This paper will describe our efforts to isolate and identify the compounds responsible for this attraction using a variety of analytic methods such as LC-QToF, GC-MS and EAG. The identified compounds will be developed as bait for a field trap for monitoring vector pressure in both Latin America and Africa. In the long term, field traps may be useful in eradication of the disease.

Why Every Hospital Should Be A “No Fly Zone”

Myiasis is the term for the invasion of living tissue by fly larvae. This occurs when a gravid fly lays its eggs in an orifice or an uncovered wound. Noninvasive wound myiasis is one of the more common forms discussed, however reports of nosocomial wound myiasis are relatively rare. This may be explained by several facts; this is not a mandatorily reportable condition, it can be interpreted as a surrogate for medical negligence, and providers may not feel it is worthy of reporting. As a result, the true frequency of myiasis cannot accurately be estimated. A case of nosocomial wound myiasis in a 55 year old male with coronary artery disease (CAD) and peripheral vascular disease (PVD) that occurred in a 750 bed university tertiary hospital was investigated.