Efficacy of Electrocuting Devices to Catch Tsetse Flies (Glossinidae) and Other Diptera

Background

The behaviour of insect vectors has an important bearing on the epidemiology of the diseases they transmit, and on the opportunities for vector control. Two sorts of electrocuting device have been particularly useful for studying the behaviour of tsetse flies (Glossina spp), the vectors of the trypanosomes that cause sleeping sickness in humans and nagana in livestock. Such devices consist of grids on netting (E-net) to catch tsetse in flight, or on cloth (E-cloth) to catch alighting flies. Catches are most meaningful when the devices catch as many as possible of the flies potentially available to them, and when the proportion caught is known. There have been conflicting indications for the catching efficiency, depending on whether the assessments were made by the naked eye or assisted by video recordings.

Methodology/Principal Findings

Using grids of 0.5m² in Zimbabwe, we developed catch methods of studying the efficiency of E-nets and E-cloth for tsetse, using improved transformers to supply the grids with electrical pulses of ~40kV. At energies per pulse of 35–215mJ, the efficiency was enhanced by reducing the pulse interval from 3200 to 1ms. Efficiency was low at 35mJ per pulse, but there seemed no benefit of increasing the energy beyond 70mJ. Catches at E-nets declined when the fine netting normally used became either coarser or much finer, and increased when the grid frame was moved from 2.5cm to 27.5cm from the grid. Data for muscoids and tabanids were roughly comparable to those for tsetse.

Conclusion/Significance

The catch method of studying efficiency is useful for supplementing and extending video methods. Specifications are suggested for E-nets and E-cloth that are ~95% efficient and suitable for estimating the absolute numbers of available flies. Grids that are less efficient, but more economical, are recommended for studies of relative numbers available to various baits.     

Copulatory Dialogues Between Male and Female Tsetse Flies (Diptera: Muscidae: Glossina pallidipes)

Traditional views of copulation and sperm transfer supposed that females are passive participants. Recent discoveries suggest, however, that females actively influence the chances that a copulation will result in fertilization of their eggs, and that they sometimes signal to males during copulation in order to elicit male responses. This paper concerns two apparent female signaling behaviors, wing vibration and body shaking, in the tsetse fly Glossina pallidipes, a species in which the male squeezes the female’s abdomen rhythmically with his powerful genitalia. Vibration was associated with male squeezes at several levels of analysis. Its coordination with male behavior suggests that vibration functions as a signal that induces the male to interrupt squeezes, but that does not forcefully dislodge the male. The female tended to vibrate her wings soon after the male began a squeeze; and when the female vibrated her wings during a squeeze, the squeeze tended to be shorter. Female body shaking was usually elicited by especially powerful squeezes. Previous studies showed that stimuli from male structures that squeeze the female probably function to induce her to ovulate, to facilitate movement of sperm into her spermathecae, and to reject the sexual advances of additional males. This study is one of the first to document an exchange of signals between male and female insects during copulation, and extends the new field of research on copulatory dialogues.     

Factors Affecting the Propensity of Tsetse Flies to Enter Houses and Attack Humans Inside: Increased Risk of Sleeping Sickness in Warmer Climates

Background

Sleeping sickness, or human African trypanosomiasis, is caused by two species of Trypanosoma brucei that are transmitted to humans by tsetse flies (Glossina spp.) when these insects take a bloodmeal. It is commonly assumed that humans must enter the normal woodland habitat of the flies to become infected, but recent studies found that tsetse frequently attack humans inside buildings. Factors affecting human/tsetse contact in buildings need identification.

Methodology/Principal Findings

In Zimbabwe, tsetse were allowed access to a house via an open door. Those in the house at sunset, and those alighting on humans in the house during the day, were caught using hand-nets. Total catches were unaffected by: (i) the presence of humans in the house and at the door, (ii) wood smoke from a fire inside the house or just outside, (iii) open windows, and (iv) chemicals simulating the odor of cattle or of humans. Catches increased about 10-fold with rising ambient temperatures, and during the hottest months the proportion of the total catch that was taken from the humans increased from 5% to 13%. Of the tsetse caught from humans, 62% consisted of female G. morsitans morstans and both sexes of G. pallidipes, i.e., the group of tsetse that normally alight little on humans. Some of the tsetse caught were old enough to be effective vectors.

Conclusion/Significance

Present results confirm previous suggestions that buildings provide a distinctive and important venue for transmission of sleeping sickness, especially since the normal repellence of humans and smoke seems poorly effective in such places. The importance of the venue would be increased in warmer climates.     

Screening of Trypanosoma brucei gambiense in Domestic Livestock and Tsetse Flies from an Insular Endemic Focus (Luba,Equatorial Guinea)

Background

Sleeping sickness is spread over 36 Sub-Saharan African countries. In West and Central Africa, the disease is caused by Trypanosoma brucei gambiense, which produces a chronic clinical manifestation. The Luba focus (Bioko Island, Equatorial Guinea) has not reported autochthonous sleeping sickness cases since 1995, but given the complexity of the epidemiological cycle, the elimination of the parasite in the environment is difficult to categorically ensure.

Methodology/Principal Findings

The aim of this work is to assess, by a molecular approach (Polymerase Chain Reaction, PCR), the possible permanence of T. b. gambiense in the vector (Glossina spp.) and domestic fauna in order to improve our understanding of the epidemiological situation of the disease in an isolated focus considered to be under control.The results obtained show the absence of the parasite in peridomestic livestock but its presence, although at very low rate, in the vector. On the other hand, interesting entomological data highlight that an elevated concentration of tsetse flies was observed in two out of the ten villages considered to be in the focus.

Conclusions

These findings demonstrate that even in conditions of apparent control, a complete parasite clearance is difficult to achieve. Further investigations must be focused on animal reservoirs which could allow the parasites to persist without leading to human cases. In Luba, where domestic livestock are scarcer than other foci in mainland Equatorial Guinea, the epidemiological significance of wild fauna should be assessed to establish their role in the maintenance of the infection.     

New Polymorphic Microsatellites in Glossina pallidipes (Diptera: Glossinidae) and Their Cross-Amplification in Other Tsetse Fly Taxa

We report the development and characterization of three new microsatellite markers in the tsetse fly, Glossina pallidipes (Diptera: Glossinidae). Fifty-eight alleles were scored in 192 individuals representing six natural populations. Allelic diversity ranged from 9 to 28 alleles per locus (mean 19.3 +/- 5.5). Averaged across loci, observed heterozygosity was 0.581 +/- 0.209, and expected heterozygosity was 0.619 +/- 0.181. Cross-species amplifications of the G. pallidipes loci in other tsetse fly taxa are reported.     

Taxonomy of Soldier Flies (Diptera: Stratiomyidae) of Sunderban Biosphere Reserve,India

The present communication deals with recording of nine species of soldier flies (Insecta: Diptera: Stratiomyidae) from Sunderban Biosphere Reserve of India, of which, Tinda indica (Walker, 1851), Hermetia illucens (Linnaeus, 1758), Odontomyia kashmirensis Brunetti, 1920, Odontomyia viridana (Wiedemann, 1824), Prosopochrysa vitripennis (Doleschall, 1856) are being reported for the first time from this unique and globally acclaimed mangrove ecosystem. Odontomyia kashmirensis Brunetti, 1920 is reported for the first time from the state of West Bengal. A key to identify these species is provided with taxonomic notes.     

The Diversity of Flower Flies (Diptera: Syrphidae) in Colombia and Their Neotropical Distribution

In Colombia, like most Neotropical countries, faunistic studies on flower flies have been occasional and most of them have been primarily focused on taxonomy. Colombia is the second-most species-rich country in flower fly diversity in the Neotropics after Brazil, and has one of the highest numbers of species per unit area (2.49 per 10,000?km²), based on a review of literature and national collections. Including new data presented here, a total of 47 genera and 300 species are recorded in Colombia. The genera Scaeva Fabricius and Lycastrirhyncha Bigot, as well as 101 species are recorded here for the first time. The altitudinal range and the distribution of the flower fly genera in Colombia are presented. A preliminary comparison of the fauna of Colombia with that of other Neotropical countries is given. A historical perspective is also provided in order to illustrate how Colombian Syrphidae knowledge has progressed over the last 168?years. Information presented here will be useful for ongoing and future biodiversity research as well as conservation projects on Syrphidae in the Neotropical region.     

The effects of host physiology on the attraction of tsetse (Diptera: Glossinidae) and Stomoxys (Diptera: Muscidae) to cattle

In Zimbabwe, studies were made of the numbers of tsetse (Glossina spp.) and stable flies (Stomoxys spp.) attracted to cattle of different nutritional status, age and sex. Host odours were analysed to determine the physiological basis of these differences and improved methods are described for measuring rates of production of kairomones. Seasonal fluctuations in host weight, related to changes in pasture quality, had no significant effect on attraction of tsetse or Stomoxys. However, both attraction to different individuals and carbon dioxide production by these individuals were strongly correlated with weight, suggesting a possible link. Attraction to the odour from different types of cattle decreased in the order ox>cow>heifer>calf, and oxen were twice as attractive as calves of less than 12 months old. Lactation did not alter the relative attractiveness of cows. Calves less than six months old produced lower levels of carbon dioxide, acetone, octenol and phenols than oxen, but for older calves and cows, levels of production of known kairomones and repellents were similar to those of an ox. Carbon dioxide produced by cattle varied according to time of day and the animal's weight; cattle weighing 500 kg produced carbon dioxide at a mean rate of 2.0 l min(-1) in the morning and 2.8 l min(-1) in the afternoon compared to respective rates of 1.1 and 1.9 l min(-1) for cattle weighing 250 kg. Artificially adjusting the doses of carbon dioxide produced by individual cattle to make them equivalent did not remove significant differences in attractiveness for tsetse but did for Stomoxys. Increasing the dose of carbon dioxide from 1 to 4 l min(-1) in a synthetic blend of identified kairomones simulating those produced by a single ox, increased attractiveness to tsetse but not to the level of an ox. The results suggest that the main sources of differences in the attractiveness of individual cattle are likely to be variation in the production of carbon dioxide and, for tsetse, other unidentified kairomone(s). The biological and practical implications of these findings are discussed.     

Characterization of microsatellite markers in the tsetse fly, Glossina pallidipes (Diptera: Glossinidae)

Glossina pallidipes is a vector of African trypanosomiasis. Here we characterize eight new polymorphic microsatellite loci in 288 G. pallidipes sampled from 12 Kenya populations. The number of alleles per locus ranged from four to 36 with a mean of 20.5 +/- 10.1. Expected single locus heterozygosities varied from 0.044 to 0.829. Heterozygosity averaged 0.616 +/- 0.246. No linkage disequilibrium was found. We also report results in eight other tsetse species estimated by using the primers developed in G. pallidipes. The primers worked best in G. swynnertoni and G. austeni and worst in G. m. morsitans and G. m. submorsitans.     

Macrogeographic population structure of the tsetse fly, Glossina pallidipes (Diptera: Glossinidae)

Tsetse flies are confined to sub-Saharan Africa where they occupy discontinuous habitats. In anticipation of area-wide control programmes, estimates of gene flow among tsetse populations are necessary. Genetic diversities were partitioned at eight microsatellite loci and five mitochondrial loci in 21 Glossina pallidipes Austin populations. At microsatellite loci, Nei's unbiased gene diversity averaged over loci was 0.659 and the total number of alleles was 214, only four of which were shared among all populations. The mean number of alleles per locus was 26.8. Random mating was observed within but not among populations (fixation index FST=0.18) and 81% of the genetic variance was within populations. Thirty-nine mitochondrial variants were detected. Mitochondrial diversities in populations varied from 0 to 0.85 and averaged 0.42, and FST=0.51. High levels of genetic differentiation were characteristic, extending even to subpopulations separated by tens and hundreds of kilometres, and indicating low rates of gene flow.     

The Effects of a Tsetse DNA Virus Infection on the Functions of the Male Accessory Reproductive Gland in the Host Fly Glossina morsitans centralis (Diptera; Glossinidae)

Freshly deposited third instar Glossina morsitans centralis larvae were infected with the tsetse DNA virus by microinjection, and at emergence adult males were separated from the females and fed on rabbit blood every second day for 8 days. A control group treated with sterile saline were handled similarly. They were dissected, and comparative observations made on the appearance and size of the accessory reproductive glands (ARG) in infected and control males. Regularly fed 8-day-old males from infected and control groups were mated to 2-day-old normal females obtained from the insectay. After separation from copula, the females were dissected and the uteri examined for the presence and quality of the spermatophore. The spermathecae were also examined for insemination. ARG tissues from the control and virus infected regularly fed 8-day-old male flies were fixed and processed for electron microscopic studies. The ARGs from control flies were found to be milky in appearance, whereas those from virus-infected flies were transparent in most parts. The ARGs from virus-infected males were significantly smaller in diameters (F = 42.26, p < 0.0001) and shorter (F = 200.4, p < 0.0001) than those of the controls. Most of the virus-infected males failed to form a complete spermatophore, whereas almost all the controls formed complete spermatophore as observed in the uteri of the female mates (Χ² = 111.661, p < 0.0001). The infected males that formed partial spermatophores and those that did not form any at all failed to inseminate their female mates. Histological studies of the ARGs revealed some lesions in the epithelial cells characterized by degeneration of cytoplasmic organelles and detachment of the muscle layer from the basal plasma membrane. However, no virus particles were observed in the affected cells. Received: 18 November 1998 / Accepted: 10 December 1998     

Eclosion behavior in tsetse (Diptera: Glossinidae): Extrication from the puparium and expansion of the adult

The tsetse adult extricates itself from the puparium and surrounding substrate by a series of muscular contractions that generate a Stereotypic pattern of changes in hemolymph pressure. The digging action of the fly can be distinguished from a second pattern of hemocoelic pulsations that is used to remove obstacles from its path. When the fly is restrained extrication behavior will persist for over 10 h. If the adult's legs are freed while the remainder of the body remains encased in the puparium, the fly fails to engage in extrication behavior, a result which suggests that freedom of the legs switches off extrication behavior and permits the onset of expansion of the body to its final adult size and shape. Expansion behavior includes walking, grooming, pumping air into the gut, and contracting the abdominal muscles to generate rhythmic pulses of hemocoelic pressure. A barographic record of internal pressure changes reflects the dynamics of this morphogenetic process. Results from tsetse are compared with previous observations recorded in flesh flies.     

The Diversity of Yellow-Related Proteins in Sand Flies (Diptera: Psychodidae)

Yellow-related proteins (YRPs) present in sand fly saliva act as affinity binders of bioamines, and help the fly to complete a bloodmeal by scavenging the physiological signals of damaged cells. They are also the main antigens in sand fly saliva and their recombinant form is used as a marker of host exposure to sand flies. Moreover, several salivary proteins and plasmids coding these proteins induce strong immune response in hosts bitten by sand flies and are being used to design protecting vaccines against Leishmania parasites. In this study, thirty two 3D models of different yellow-related proteins from thirteen sand fly species of two genera were constructed based on the known protein structure from Lutzomyia longipalpis. We also studied evolutionary relationships among species based on protein sequences as well as sequence and structural variability of their ligand-binding site. All of these 33 sand fly YRPs shared a similar structure, including a unique tunnel that connects the ligand-binding site with the solvent by two independent paths. However, intraspecific modifications found among these proteins affects the charges of the entrances to the tunnel, the length of the tunnel and its hydrophobicity. We suggest that these structural and sequential differences influence the ligand-binding abilities of these proteins and provide sand flies with a greater number of YRP paralogs with more nuanced answers to bioamines. All these characteristics allow us to better evaluate these proteins with respect to their potential use as part of anti-Leishmania vaccines or as an antigen to measure host exposure to sand flies.     

Management of Fruit Flies (Diptera: Tephritidae) of the Most Perishable Fruits

We investigated to minimize the dependency on the use of chemicals and thus develop safe and environmental friendly control program for the most perishable fruits i.e., apple,‘ber’, guava and mango. Our findings on the composition of fruit fly species reveal that Bactrocera dorsalis was dominant on apple (33.96% existence), Corpomya incompleta on‘ber’(51.91% existence) and Bactrocera zonata on guava (49.62% existence) and mango (74.66% existence). The correlation between population and infestation percentage was non‐significant in apple orchards, whereas positive and highly significant in between population and infestation, as well as on the cumulative basis in‘ber’, guava and mango orchards during 1998‐1999. Hoeing, baiting and methyl eugenol were statistically equal resulting about 77% decrease in infestation. The maximum control of 91.68% was observed where all four‐control operations including Dipterex® were integrated together. Weather factors, when computed together, had maximum effect on population fluctuation and infestation with rainfall contributing the major role. For guava fruits, the months of August (14.06A individuals/trap/day) and September (13.81A individuals/trap/day) were important, resulting in maximum infestation percentage of 10.76 to 14.74%, respectively.     

Maturation of the tsetse fly Glossina pallidipes (Diptera: Glossinidae) in relation to trap-orientated behaviour

ABSTRACT The fat free dry weight or residual dry weight of the thorax (Trdw) increased linearly for the first 10 days of adult life in both sexes of G.pallidipes in the laboratory as their flight muscles developed. Using ovarian dissection to estimate the ages of nulliparous adult females of G.pallidipes , the Trdw was also found to increase linearly for at least 14 days in the field. Significant increases in pteridine fluorescence with age were measured in both laboratory-reared males and females of known chronological age and in wild-caught nulliparous females whose ages were estimated by ovarian dissection. A linear relationship existed between pteridine fluorescence and wing fray category for a wide range of ages of field-caught flies of both sexes. A stationary trap baited with ox odour was selective in that only the hungrier portion of the flies attracted to it actually entered. However, it was not selective in terms of the mean ages of flies caught. Comparisons were made of the age compositions of catches of both sexes of G.pallidipes attracted to a stationary trap baited with synthetic odours or to a mobile electrified net by plotting Trdw values against pteridine fluorescence. Nulliparous females were not attracted to the stationary trap, but were attracted to the mobile bait. Males of all ages appeared to be equally attracted to both. It is concluded that nulliparous females do not respond to host odour stimuli until they are ready to mate, perhaps relying on the energy-conserving strategy of watching for a moving host animal before attempting to feed. Alternatively, synthetic odours may differ from natural host odours in terms of their attractiveness to young females. Males, however, probably exhibit dual sexual and feeding behaviour by responding to an odour-baited stationary trap even when young.     

Functional Morphology of the Phallosome in Glossina (Diptera, Glossinidae) and Its Evolutionary Implications

Current views on the phylo-genetjc relations of the three species groups within the genus Glossina are critically reviewed. In respect of the male genitalia, the fusca group is here regarded as more specialized than the morsitans group. The detailed phallosome morphology of G. austeni is compared with that of G. fuscipes , on the basis of their mode of action in living flies. The comparison is extended to fusca group species, using dead material only. The presumed phylogeny of Glossina is presented in outline; the evolution of the genus has involved a progressive invasion of riverine and later of forest habitats, from an original semi-arid grassy woodland habitat.     

Modelling the effect of feeding-related mortality on the feeding strategy of tsetse (Diptera: Glossinidae)

Abstract. Free-living haematophagous insects risk death through host grooming responses or through increased susceptibility to predation whenever they take a bloodmeal. In this paper we investigate the effects of these risks on the feeding strategy of tsetse. A model is presented that allows for death of tsetse by starvation if they do not succeed in feeding within a fixed time (set at 6 days in the first instance) and for mortality specifically associated with feeding. In addition there is background mortality that applies to all flies at all times.
The model is used to compute the individual life-time fertility (number of female puparia per female) as a function of the probability of obtaining a meal (indicated by field data to be very high, usually > 0.85 per day) and the day on which flies start to search for a meal. We suggest that the feeding strategy that would be selected for is that which allows the maximum reproductive output. The model shows that this strategy involves making no attempts to feed for 3–4 days after the previous meal and then attempting to feed with the greatest possible probability until a meal is obtained. The predicted feeding interval, obtained independently of any trapping data, agrees closely with all previous estimates from field studies using a variety of methods. Preliminary results from a laboratory experiment reveal an increased risk of predation of recently fed as compared with hungry tsetse. The lower the actual feeding mortality the more frequently will flies be able to feed should conditions so demand. It is adaptive, however, for tsetse to delay attempting to feed for as long as they can, which is made possible by the near certainty of locating and feeding on a host within 1 day, using their sophisticated sensory systems.     

Australian Distribution of 17 Species of Fruit Flies (Diptera: Tephritidae) Caught in Cue Lure Traps in February 1994

We report the first widespread survey of tephritid fruit flies attempted in a single time period. 1,471 cue lure traps caught 17 species, and extensions to previously recorded geographical ranges were detected for seven of them: Bactrocera tryoni, B. neohumeralis, B. frauenfeldi, B. aeroginosa, Dacus absonifascies, D. aequalis and D. newmani. the traps also unexpectedly caught several B. cacuminata and also both males and females of Dirioxa pornia and Ceratitis capitata. the geographical variation in the relative abundance of B. tryoni and B. neohumeralis in the region of their co-occurrence was in substantial agreement with earlier estimates. the regional variation in abundance of B. tryoni in the eastern states was in accordance with the predictions of a published bioclimatic model. Furthermore, the spread of this species (expected from the model) to several locations in the Northern Territory is recorded here for the first time.