The response of tsetse flies to artificial baits in relation to age, nutritional and reproductive state

In various vegetation types in Zimbabwe, the catches of Glossina pallidipes Austen and G. morsitans morsitans Westw. (Diptera: Glossinidae) at a target baited with odour (acetone, 1-octen-3-ol and two phenols) were positively correlated with catches of the same species at an unbaited net. No correlation existed between target catches and hand net catches of tsetse flies sitting on the vegetation. G. pallidipes females caught at a target and at an unbaited net were older than those caught from vegetation. Of the female G. pallidipes caught at the target, 46% were in the first 3 days of pregnancy. Of those caught at the unbaited net, significantly fewer, 21%, were in this stage. G. pallidipes males caught from vegetation contained more fat (3.07±0.333 mg) than those caught at the unbaited net (2.06±0.339 mg) or at the target (2.19±0.218 mg). It is inferred that target catches consisted predominantly of tsetse which were already in flight when they sensed the stimuli from the target, and that target catches were biased towards female G. pallidipes in the first 3 days of pregnancy.     

Characterization of a composite with enhanced attraction to savannah tsetse flies from constituents or analogues of tsetse refractory waterbuck (Kobus defassa) body odor

Savannah tsetse flies avoid flying toward tsetse fly-refractory waterbuck (Kobus defassa) mediated by a repellent blend of volatile compounds in their body odor comprised of δ-octalactone, geranyl acetone, phenols (guaiacol and carvacrol), and homologues of carboxylic acids (C₅-C₁₀) and 2-alkanones (C₈-C₁₃). However, although the blends of carboxylic acids and that of 2-alkanones contributed incrementally to the repellency of the waterbuck odor to savannah tsetse flies, some waterbuck constituents (particularly, nonanoic acid and 2-nonanone) showed significant attractive properties. In another study, increasing the ring size of δ-octalactone from six to seven membered ring changed the activity of the resulting molecule (ε-nonalactone) on the savannah tsetse flies from repellency to attraction. In the present study, we first compared the effect of blending ε-nonalactone, nonanoic acid and 2-nonanone in 1:1 binary and 1:1:1 ternary combination on responses of Glossina pallidipes and Glossina morsitans morsitans tsetse flies in a two-choice wind tunnel. The compounds showed clear synergistic effects in the blends, with the ternary blend demonstrating higher attraction than the binary blends and individual compounds. Our follow up laboratory comparisons of tsetse fly responses to ternary combinations with different relative proportions of the three components showed that the blend in 1:3:2 proportion was most attractive relative to fermented cow urine (FCU) to both tsetse species. In our field experiments at Shimba Hills game reserve in Kenya, where G. pallidipes are dominant, the pattern of tsetse catches we obtained with different proportions of the three compounds were similar to those we observed in the laboratory. Interestingly, the three-component blend in 1:3:2 proportion when released at optimized rate of 13.71mg/h was 235% more attractive to G. pallidipes than a combination of POCA (3-n-Propylphenol, 1-Octen-3-ol, 4-Cresol, and Acetone) and fermented cattle urine (FCU). This constitutes a novel finding with potential for downstream deployment in bait technologies for more effective control of G. pallidipes, G. m. morsitans, and perhaps other savannah tsetse fly species, in ‘pull’ and ‘pull-push’ tactics.     

Age‐specific changes in sperm levels among female tsetse (Glossina spp.) with a model for the time course of insemination

The age, insemination and ovulation status of tsetse flies Glossina pallidipes Austen (n = 154369) and Glossina morsitans morsitans Westwood (n = 19659), captured over 11 years in Zimbabwe, are assessed by ovarian dissection. Instantaneous rates of insemination increase exponentially with age in both species; 90% insemination levels are reached after 5 days post‐emergence in G. m. morsitans and 7 days in G. pallidipes, varying little with season. More than 95% of both species have ovulated by the age of 8 days and 99% by 12 days. Older flies that have not ovulated are > 100‐fold more likely to be caught in October and November than in other months. A 500‐fold decrease in trap catches did not result in any detectible decrease in the probability of females being inseminated. The proportion of partially filled spermathecae rises for approximately 6 days then declines, consistent with some flies having mated more than once. For flies caught on electric nets, with wings undamaged during capture, wing‐fray data are used to extend ovarian age estimates up to 11 ovulations. Among these flies, the volume of sperm in the spermathecae declines little in flies that have ovulated up to seven times; thereafter, it declines by approximately 1% per ovulation. The time course of insemination and the mating frequency of females are important considerations in modelling tsetse fly populations, as well as for the dynamics of interventions involving the release of genetically‐modified insects, which should not be seriously compromised by the limited levels of polyandry currently observed.     

Do tsetse flies ‘see’ zebras? A field study of the visual response of tsetse to striped targets

Abstract A field study in Zimbabwe of Glossina pallidipes Austen and G. morsitans morsitans Westwood supported Waage's (1981) hypothesis that the striped pattern of zebras may protect them from being bitten by blood-sucking flies. In addition, the results suggest that the orientation of the stripes may be crucially important for the unattractiveness of zebras. The relative attractiveness of five different stationary targets (black, white, grey, vertically-striped and horizontally-striped; stripe width = 5 cm) were each tested on their own and in pairs of all combinations, with artificial host odour (CO₂ plus acetone) always present. Electric nets were used to catch flies as they attempted to land on or circle the targets. The results were similar for the two species of tsetse. When tested on their own, grey and vertically-striped targets caught similar numbers of flies and both caught significantly fewer than black or white targets (c. 36% as many). Horizontally-striped targets caught <10% as many flies as any other single target. Although there was no significant difference between the attractiveness of grey and vertically striped targets when they were presented together, when paired with the other targets, grey was as attractive as black or white, but the vertically-striped target was significantly less attractive than black or white (P < 0.001). In other words, a difference between grey and vertical stripes was found only in their attractiveness in relation to other targets. The horizontally-striped target, however, always caught the fewest flies, regardless of whether it was presented alone or alongside another target.     

A comparison of susceptibility of two allopatric populations of Glossina pallidipes for stocks of Trypanosoma congolense

Abstract. A colony of Glossina pallidipes Austen which originated from Nguruman, Rift Valley Province, Kenya, was significantly more susceptible to infection (19.3%) with a stock of Trypanosoma congolense Broden isolated from G. pallidipes in Nguruman than a colony of the same species which originated from Shimba Hills, Coast Province, Kenya (5.6%). Male G. pallidipes from Nguruman were significantly more susceptible than females to this T. congolense stock whilst the susceptibility of both sexes of G. pallidipes from Shimba Hills did not differ significantly. All six goats on which six infected G. pallidipes fed singly (three tsetse per colony) became infected. Similarly, the G. pallidipes colony of Nguruman origin was significantly more susceptible to infection (16.4%) with a stock of T. congolense isolated from G. pallidipes in Shimba Hills than the colony of Shimba Hills origin (4.9%). The susceptibility of the sexes of G. pallidipes from both the colonies to this stock of T. congolense did not differ significantly. Again, all six goats on which six infected G. pallidipes fed singly (three tsetse per colony) became infected. If the observed differences in susceptibility of the two G. pallidipes colonies reflect transmission of trypanosomes by the two allopatric populations of tsetse in the field, then the epidemiology of congolense-trypanosomiasis in livestock must differ between these two areas of Kenya endemic for trypanosomiasis.     

Salivary gland hyperplasia in wild caught tsetse from Zimbabwe

Hypertrophied salivary glands were collected from wild populations of the tsetse flies Glossina morsitans morsitans Westwood and G. pallidipes Austen in the Zambezi valley, Zimbabwe. Examination of the glands by electron microscopy showed that the associated virus was identical in form in both species. The incidence of salivary gland hypertrophy was significantly higher in G. pallidipes than in G.m.morsitans, and 83% of the enlarged glands of the former were, in addition, found to be infected with Rickettsia-like organisms. There was no indication from this small sample that viral infections predisposed flies to infections with trypanosomes.

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Résumé Des glandes salivaires hypertrophiées ont été extraites de Glossina morsitans et de G. pallidipes provenant de populations sauvages capturées dans la vallée du Zambèze au Zimbabwe. L'examen des glandes au microscope électronique a montré que la forme du virus associé était identique dans les deux espèces. La fréquence d'hypertrophie des glandes salivaires était significativement plus élevée chez G. pallidipes que chez G. morsitans et la fréquence des glandes hypertrophiées de G. pallidides contaminèes par des organismes ressemblant à des Rickettsia était de 0,02. Rien n'indique à partir de ce petit échantillon que la contamination virale favorise l'infection par des trypanosomes.

     

Factors affecting density-independent survival of an island population of tsetse flies in Zimbabwe

Analysis is presented of the factors affecting survival probability in populations of tsetse flies Glossina morsitans morsitans Westwood and G. pallidipes Austen (Diptera: Glossinidae) on Antelope Island, Lake Kariba, Zimbabwe. For mature male and female adult G. m. morsitans mean temperature (T _bar) accounted for 70 and 50%, respectively, of the variance in mark-recapture estimates of survival when the flies were not subjected to trapping. Saturation deficit (SD) only accounted for 36 and 33%, respectively. Maximum temperature (T_max) and SD accounted for 36–42% of the variance in male and female G. pallidipes. For the corresponding Moran curve estimates of the survival over all developmental stages, SD lagged by three weeks accounted for 61 and 41% of the variance for male and female G. m. morsitans, respectively, and 64 and 56% for G. pallidipes. The corresponding figures for plots against T_max were 44, 23, 23, and 21%, respectively. The same patterns were seen in the whole data set once allowance was made for the effect of trapping on survival and for an effect of season, correlated with an index of photosynthetic activity. For male G. m. morsitans there was a significant effect of saturation deficit, but not temperature, on immature survival. Decreased adult survival at high temperatures results from the need to feed more frequently and hence to take more risks per unit time. High saturation deficits result directly in reduced emergence of healthy flies from pupae.     

Seasonal variation of tsetse fly species abundance and prevalence of trypanosomes in the Maasai Steppe,Tanzania

Tsetse flies, the vectors of trypanosomiasis, represent a threat to public health and economy in sub‐Saharan Africa. Despite these concerns, information on temporal and spatial dynamics of tsetse and trypanosomes remain limited and may be a reason that control strategies are less effective. The current study assessed the temporal variation of the relative abundance of tsetse fly species and trypanosome prevalence in relation to climate in the Maasai Steppe of Tanzania in 2014–2015. Tsetse flies were captured using odor‐baited Epsilon traps deployed in ten sites selected through random subsampling of the major vegetation types in the area. Fly species were identified morphologically and trypanosome species classified using PCR. The climate dataset was acquired from the African Flood and Drought Monitor repository. Three species of tsetse flies were identified: G. swynnertoni (70.8%), G. m. morsitans (23.4%), and G.pallidipes (5.8%). All species showed monthly changes in abundance with most of the flies collected in July. The relative abundance of G. m. morsitans and G. swynnertoni was negatively correlated with maximum and minimum temperature, respectively. Three trypanosome species were recorded: T. vivax (82.1%), T. brucei (8.93%), and T. congolense (3.57%). The peak of trypanosome infections in the flies was found in October and was three months after the tsetse abundance peak; prevalence was negatively correlated with tsetse abundance. A strong positive relationship was found between trypanosome prevalence and temperature. In conclusion, we find that trypanosome prevalence is dependent on fly availability, and temperature drives both tsetse fly relative abundance and trypanosome prevalence.     

Responses of the tsetse fly, Glossina pallidipes, to ox odour, carbon dioxide and a visual stimulus in the laboratory

The responses of male and female Glossina pallidipes Austen to a visual target were recorded in a slow-speed wind tunnel, using a video system. Addition of ox odour or carbon dioxide at an equivalent concentration to the airstream resulted in an increase in flight activity and a marked increase in flies alighting on the visual target. In the absence of ox odour flights were characterised by a number of collisions with the walls and ceiling of the cage used to retain the flies, whereas in the presence of ox odour the flies circled around the centre of the cage avoiding the edges. Removal of the visual target did not alter this response. The results are discussed in the light of field observations on the behaviour of G. pallidipes around baited targets. When flies were observed in groups, mutual disturbance increased the activity during control periods thus masking the activating effect of ox odour. The activity of individual flies occurred in bursts (22.1 s mean duration) consisting of a number of flights (3.3 s mean duration) and longer periods of inactivity (85.8 s mean duration). The burst lenght did not change when ox odour was added to the airstream but the number of flights per burst increased. These results are discussed in relation to the random dispersal theory of tsetse populations.

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Résumé Les réactions de G. pallidipes mâles et femelles, à l'odeur de vache, au gaz carbonique et à des stimulations visuelles ont été enregittrées sur un système vidéo, lors d'expériences dans un tunnel à vent à faible vitesse. Les adultes ont été mis à jeûner pendant 3 jours après leur premier repas, avant d'être introduits à l'intérieur du tunnel à vent dans une cage en fil nylon, conçue pour maintenir les mouches dans le champ de la caméra. Dans les premières expériences, 10 mouches ont été observées simultanément. L'introduction d'odeur de vache ou de CO₂ n'a augmenté l'activité d'aucun des deux sexes, mais a brutalement accru le nombre de vols s'achevant sur une cible mate noire. Une étude plus précise et continue pendant 60 minutes, de groupes de 5 mâles et de 5 femelles, a révélé que les envols n'étaient pas indépendants et que les perturbations mutuelles avaient masqué l'action stimulante de l'odeur de vache et du CO₂. Pour surmonter le problème des perturbations mutuelles, les mouches ont été observées individuellement. Mâles et femelles ont répondu par une augmentation de l'activité de vol à l'introduction d'odeur de vache dans le courant d'air. Les mâles ne se sont posés sur les cibles que dans les minutes ayant suivi l'addition d'odeur de vache, tandis que les femelles se sont posées tout le long de l'expérience, le nombre d'atterrissages s'étant accru significativement avec l'addition d'odeur de vache. L'odeur de vache a modifié le type de vol, passant du vol au hasard entraînant de nombreuses collisions avec les parois et le plafond de la cage, à un vol plus direct provoquant le tournoiement des mouches dans la cage. Par ailleurs, ce comportement a subsisté après le retrait des cibles mates noires de la cage. On peut en conclure que l'odeur de vache active les femelles et augmente la perception visuelle du paysage. L'activité de G. pallidipes avait lieu avec de brêves poussées (durée moyenne 22,1 secondes) de nombreux vols, suivies de périodes d'inactivité plus longues (durée moyenne 85,8 secondes). La durée moyenne d'un vol était de 3,3 s. Partant de ces données et d'une vitesse de vol estimée à 5,0 m/s, la longeur d'un vol élémentaire pour une dispersion théoriquement au hasard, est 16,5 m. La durée de la poussée d'activité n'a pas changé avec l'introduction d'odeur de vache, mais le nombre de vols par poussée a augmenté, et bien qu'il ait été impossible de comparer la durée des périodes de repos avec et sans odeur, des expériences précédentes suggèrent qu'elles devraient diminuer en présence d'odeur.

     

Analysis of the gut-specific microbiome from field-captured tsetse flies,and its potential relevance to host trypanosome vector competence

Background

The tsetse fly (Glossina sp.) midgut is colonized by maternally transmitted and environmentally acquired bacteria. Additionally, the midgut serves as a niche in which pathogenic African trypanosomes reside within infected flies. Tsetse’s bacterial microbiota impacts many aspects of the fly’s physiology. However, little is known about the structure of tsetse’s midgut-associated bacterial communities as they relate to geographically distinct fly habitats in east Africa and their contributions to parasite infection outcomes. We utilized culture dependent and independent methods to characterize the taxonomic structure and density of bacterial communities that reside within the midgut of tsetse flies collected at geographically distinct locations in Kenya and Uganda.

Results

Using culture dependent methods, we isolated 34 strains of bacteria from four different tsetse species (G. pallidipes, G. brevipalpis, G. fuscipes and G. fuscipleuris) captured at three distinct locations in Kenya. To increase the depth of this study, we deep sequenced midguts from individual uninfected and trypanosome infected G. pallidipes captured at two distinct locations in Kenya and one in Uganda. We found that tsetse’s obligate endosymbiont, Wigglesworthia, was the most abundant bacterium present in the midgut of G. pallidipes, and the density of this bacterium remained largely consistent regardless of whether or not its tsetse host was infected with trypanosomes. These fly populations also housed the commensal symbiont Sodalis, which was found at significantly higher densities in trypanosome infected compared to uninfected flies. Finally, midguts of field-captured G. pallidipes were colonized with distinct, low density communities of environmentally acquired microbes that differed in taxonomic structure depending on parasite infection status and the geographic location from which the flies were collected.

Conclusions

The results of this study will enhance our understanding of the tripartite relationship between tsetse, its microbiota and trypanosome vector competence. This information may be useful for developing novel disease control strategies or enhancing the efficacy of those already in use.

     

Comment on Barclay and Vreysen: Published dynamic population model for tsetse cannot fit field data

The structure, and assumed parameter values, of a recent dynamic population model for tsetse (Diptera: Glossinidae) render it unable to fit published data on tsetse control programs using odor-baited targets, insecticide-treated cattle and the sterile insect technique (SIT). The underlying problem is a mismatch between the small size of the mapped cells (1 ha) and the long time-step, which allows flies to move only once every 5 days, and then only to an adjacent cell. Assumed rates of tsetse dispersal and killing by odor-baited targets are consequently at least an order of magnitude lower than observed in the field. Suggestions that Glossina pallidipes could be eradicated more rapidly with SIT, than using hundreds of targets per km², is contradicted both by the field data and by three other independent modeling studies.     

The nutritional state of male tsetse flies,Glossina pallidipes,at the time of feeding

Abstract. The feeding intervals of tsetse flies have been estimated from the nutritional state of flies caught in traps. However, such estimates have been disputed on the grounds that traps catch a biased, hungry sample of the flies which are seeking hosts and will feed. In this paper we present data on the nutritional state of tsetse flies caught approaching and feeding on oxen. Individual oxen were surrounded with an incomplete ring of electric nets which caught Glossina pallidipes Austen that were approaching, departing unfed and departing fed from an ox. Non-teneral males caught in this way were analysed for their fat and haematin contents. The feeding interval was estimated from a comparison of the frequency distributions of the pre- and post-feed haematin contents of the flies which fed. The former was not measured directly, and was deduced from the frequency distributions of the haematin contents of the male flies caught approaching and departing unfed from the oxen, since it is assumed that the departing unfed and fed flies together form a sample of the approaching flies. There was no difference between the frequency distributions of haematin contents of flies caught approaching and departing unfed, and therefore the pre-feed haematin contents of the males which fed should have the same frequency distribution. Comparison of this distribution with that of the post-feed haematin contents of the males which fed indicated that the majority of male G.pallidipes were returning to feed after digesting on average 1.4 log haematin units of the previous bloodmeal. From data published elsewhere, this corresponds to a mean feeding interval of 42-60h. There was a strong, linear, negative relationship between the fat contents of males and their probability of taking a bloodmeal, suggesting that fat content is important in determining the feeding behaviour of tsetse flies.     

Prospects for the Development of Odour Baits to Control the Tsetse Flies Glossina tachinoides and G. palpalis s.l.

Field studies were done of the responses of Glossina palpalis palpalis in Côte d''Ivoire, and G. p. gambiensis and G. tachinoides in Burkina Faso, to odours from humans, cattle and pigs. Responses were measured either by baiting (1.) biconical traps or (2.) electrocuting black targets with natural host odours. The catch of G. tachinoides from traps was significantly enhanced (∼5×) by odour from cattle but not humans. In contrast, catches from electric targets showed inconsistent results. For G. p. gambiensis both human and cattle odour increased (>2×) the trap catch significantly but not the catch from electric targets. For G. p. palpalis, odours from pigs and humans increased (∼5×) the numbers of tsetse attracted to the vicinity of the odour source but had little effect on landing or trap-entry. For G. tachinoides a blend of POCA (P = 3-n-propylphenol; O = 1-octen-3-ol; C = 4-methylphenol; A = acetone) alone or synthetic cattle odour (acetone, 1-octen-3-ol, 4-methylphenol and 3-n-propylphenol with carbon dioxide) consistently caught more tsetse than natural cattle odour. For G. p. gambiensis, POCA consistently increased catches from both traps and targets. For G. p. palpalis, doses of carbon dioxide similar to those produced by a host resulted in similar increases in attraction. Baiting traps with super-normal (∼500 mg/h) doses of acetone also consistently produced significant but slight (∼1.6×) increases in catches of male flies. The results suggest that odour-baited traps and insecticide-treated targets could assist the AU-Pan African Tsetse and Trypanosomiasis Eradication Campaign (PATTEC) in its current efforts to monitor and control Palpalis group tsetse in West Africa. For all three species, only ∼50% of the flies attracted to the vicinity of the trap were actually caught by it, suggesting that better traps might be developed by an analysis of the visual responses and identification of any semiochemicals involved in short-range interaction.     

Field experiments on a new method for determining age in tsetse flies (Diptera: Glossinidae)

ABSTRACT.

• 1 Data are presented which suggests that the accurate determination of the age of tsetse flies (Glossina morsitans morsitans Westwood and G. pallidipes Austen) in the field can be achieved by measuring the fluorescence content of the head capsule.
• 2 The way in which this can be achieved and further work which would improve the accuracy of the technique are discussed.

     

Three‐dimensional flight tracking shows how a visual target alters tsetse fly responses to human breath in a wind tunnel

Tsetse flies Glossina spp. (Diptera; Glossinidae) are blood‐feeding vectors of disease that are attracted to vertebrate hosts by odours and visual cues. Studies on how tsetse flies approach visual devices are of fundamental interest because they can help in the development of more efficient control tools. The responses of a forest tsetse fly species Glossina brevipalpis (Newstead) to human breath are tested in a wind tunnel in the presence or absence of a blue sphere as a visual target. The flight responses are video recorded with two motion‐sensitive cameras and characterized in three dimensions. Although flies make meandering upwind flights predominantly in the horizontal plane in the plume of breath alone, upwind flights are highly directed at the visual target presented in the plume of breath. Flies responding to the visual target fly from take‐off within stricter flight limits at lower ground speeds and with a significantly lower variance in flight trajectories in the horizontal plane. Once at the target, flies fly in loops principally in the horizontal plane within 40 cm of the blue sphere before descending in spirals beneath it. Successful field traps designed for G. brevipalpis take into account the strong horizontal component in local search behaviour by this species at objects. The results suggest that trapping devices should also take into account the propensity of G. brevipalpis to descend to the lower parts of visual targets.     

Flower and fruit volatiles assist host‐plant location in the Tomato fruit fly Neoceratitis cyanescens

Mature females of the tomato fruit fly Neoceratitis cyanescens can detect host fruit at a short distance using only visual stimuli, but little is known about the role of airborne volatile cues in the host searching strategy. A series of experiments is conducted in a laboratory wind tunnel, in which the behavioural responses of individual flies to volatiles from Solanaceae host plants (including tomato Lycopersicum esculentum Mill., bug weed Solanum mauritianum Scop. and Turkey berry Solanum torvum Sw.) are observed, according to some environmental (air speed) and physiological (age and mating status of females, time of day) factors. Mature females respond primarily to specific olfactory cues from blends of flowers or host fruit, preferentially unripe fruit for bug weed, as opposed to ripe fruit for Turkey berry or tomato. Males are also highly attracted by the odour of unripe fruit of bug weed. Wind plays a key role, as shown by the proportion of flies that reach the upwind section of the tunnel in the presence of both fruit odour and air flow (66.7%) and in the absence of either fruit odour (13.3%) or wind (36.7%). In response to fruit volatiles carried by wind, flies embark in a ‘plume tracking’ or ‘aim and shoot' flight, consistent with odour‐conditioned anemotaxis. Females respond to host fruit odour regardless of their age, egg load or mating status, and also more consistently in the afternoon, which is their preferential time of day for egg‐laying. Searching behaviour and response to host volatiles in N. cyanescens are discussed in the light of host‐finding and an adaptive strategy.     

Prospects for Developing Odour Baits To Control Glossina fuscipes spp., the Major Vector of Human African Trypanosomiasis

We are attempting to develop cost-effective control methods for the important vector of sleeping sickness, Glossina fuscipes spp. Responses of the tsetse flies Glossina fuscipes fuscipes (in Kenya) and G. f. quanzensis (in Democratic Republic of Congo) to natural host odours are reported. Arrangements of electric nets were used to assess the effect of cattle-, human- and pig-odour on (1) the numbers of tsetse attracted to the odour source and (2) the proportion of flies that landed on a black target (1×1 m). In addition responses to monitor lizard (Varanus niloticus) were assessed in Kenya. The effects of all four odours on the proportion of tsetse that entered a biconical trap were also determined. Sources of natural host odour were produced by placing live hosts in a tent or metal hut (volumes≈16 m³) from which the air was exhausted at ∼2000 L/min. Odours from cattle, pigs and humans had no significant effect on attraction of G. f. fuscipes but lizard odour doubled the catch (P<0.05). Similarly, mammalian odours had no significant effect on landing or trap entry whereas lizard odour increased these responses significantly: landing responses increased significantly by 22% for males and 10% for females; the increase in trap efficiency was relatively slight (5–10%) and not always significant. For G. f. quanzensis, only pig odour had a consistent effect, doubling the catch of females attracted to the source and increasing the landing response for females by ∼15%. Dispensing CO₂ at doses equivalent to natural hosts suggested that the response of G. f. fuscipes to lizard odour was not due to CO₂. For G. f. quanzensis, pig odour and CO₂ attracted similar numbers of tsetse, but CO₂ had no material effect on the landing response. The results suggest that identifying kairomones present in lizard odour for G. f. fuscipes and pig odour for G. f. quanzensis may improve the performance of targets for controlling these species.     

Sodalis glossinidius presence in wild tsetse is only associated with presence of trypanosomes in complex interactions with other tsetse-specific factors

Background

Susceptibility of tsetse flies (Glossina spp.) to trypanosomes of both humans and animals has been associated with the presence of the endosymbiont Sodalis glossinidius. However, intrinsic biological characteristics of the flies and environmental factors can influence the presence of both S. glossinidius and the parasites. It thus remains unclear whether it is the S. glossinidius or other attributes of the flies that explains the apparent association. The objective of this study was to test whether the presence of Trypanosoma vivax, T. congolense and T. brucei are related to the presence of S. glossinidius in tsetse flies when other factors are accounted for: geographic location, species of Glossina, sex or age of the host flies.

Results

Flies (n = 1090) were trapped from four sites in the Shimba Hills and Nguruman regions in Kenya. Sex and species of tsetse (G. austeni, G. brevipalpis, G. longipennis and G. pallidipes) were determined based on external morphological characters and age was estimated by a wing fray score method. The presence of trypanosomes and S. glossinidius was detected using PCR targeting the internal transcribed spacer region 1 and the haemolysin gene, respectively. Sequencing was used to confirm species identification. Generalised Linear Models (GLMs) and Multiple Correspondence Analysis (MCA) were applied to investigate multivariable associations. The overall prevalence of trypanosomes was 42.1%, but GLMs revealed complex patterns of associations: the presence of S. glossinidius was associated with trypanosome presence but only in interactions with other factors and only in some species of trypanosomes. The strongest association was found for T. congolense, and no association was found for T. vivax. The MCA also suggested only a weak association between the presence of trypanosomes and S. glossinidius. Trypanosome-positive status showed strong associations with sex and age while S. glossinidius-positive status showed a strong association with geographic location and species of fly.

Conclusions

We suggest that previous conclusions about the presence of endosymbionts increasing probability of trypanosome presence in tsetse flies may have been confounded by other factors, such as community composition of the tsetse flies and the specific trypanosomes found in different regions.

     

Hytrosavirus genetic diversity and eco-regional spread in Glossina species

Background

The management of the tsetse species Glossina pallidipes (Diptera; Glossinidae) in Africa by the sterile insect technique (SIT) has been hindered by infections of G. pallidipes production colonies with Glossina pallidipes salivary gland hypertrophy virus (GpSGHV; Hytrosaviridae family). This virus can significantly decrease productivity of the G. pallidipes colonies. Here, we used three highly diverged genes and two variable number tandem repeat regions (VNTRs) of the GpSGHV genome to identify the viral haplotypes in seven Glossina species obtained from 29 African locations and determine their phylogenetic relatedness.

Results

GpSGHV was detected in all analysed Glossina species using PCR. The highest GpSGHV prevalence was found in G. pallidipes colonized at FAO/IAEA Insect Pest Control Laboratory (IPCL) that originated from Uganda (100%) and Tanzania (88%), and a lower prevalence in G. morsitans morsitans from Tanzania (58%) and Zimbabwe (20%). Whereas GpSGHV was detected in 25–40% of G. fuscipes fuscipes in eastern Uganda, the virus was not detected in specimens of neighboring western Kenya. Most of the identified 15 haplotypes were restricted to specific Glossina species in distinct locations. Seven haplotypes were found exclusively in G. pallidipes. The reference haplotype H1 (GpSGHV-Uga; Ugandan strain) was the most widely distributed, but was not found in G. swynnertoni GpSGHV. The 15 haplotypes clustered into three distinct phylogenetic clades, the largest contained seven haplotypes, which were detected in six Glossina species. The G. pallidipes-infecting haplotypes H10, H11 and H12 (from Kenya) clustered with H7 (from Ethiopia), which presumably corresponds to the recently sequenced GpSGHV-Eth (Ethiopian) strain. These four haplotypes diverged the most from the reference H1 (GpSGHV-Uga). Haplotypes H1, H5 and H14 formed three main genealogy hubs, potentially representing the ancestors of the 15 haplotypes.

Conclusion

These data identify G. pallidipes as a significant driver for the generation and diversity of GpSGHV variants. This information may provide control guidance when new tsetse colonies are established and hence, for improved management of the virus in tsetse rearing facilities that maintain multiple Glossina species.

     

Tsetse flies are attracted to the invasive plant Lantana camara

In tsetse both sexes feed exclusively on the blood of vertebrates for a few minutes every 2-3 days. Tsetse flies seek cover from high temperatures to conserve energy and plants provide shelter for tsetse in all the biotopes they occupy. Recently, tsetse have taken cover in plantations and under the invasive bush Lantana camara that has invaded large areas of the tsetse fly belt of Africa. Flies from such refugia are implicated in sleeping sickness epidemics. In a wind tunnel we show that both foliage and an extract of volatiles from foliage of L. camara attract three tsetse spp. from different habitats: Glossina fuscipes fuscipes (riverine), G. brevipalpis (sylvatic) and G. pallidipes (savannah). Gas chromatography analysis of volatiles extracted from leaves and flowers of L. camara coupled to electroantennograme recordings show that 1-octen-3-ol and beta-caryophyllene are the major chemostimuli for the antennal receptor cells of the three tsetse spp. studied. A binary mixture of these products attracted these flies in the wind tunnel. The gas chromatography linked electroantennograme analysis of the L. camara extracts also show that the antennal receptor cells of the three tsetse spp. respond similarly to groups of volatiles derived from the major biosynthetic and catabolic pathways of plants, i.e. to mono- and sesquiterpenes, to lipoxidation products and to aromatics. Mixtures of these plant volatiles also attracted tsetse in the wind tunnel. These findings show that tsetse flies have conserved a strong sensitivity to volatile secondary products of plants, underlining the fundamental role of vegetation in tsetse survival.