Behaviour of tsetse flies (Glossina) in host odour plumes in the field

ABSTRACT. In Zimbabwe, studies were made of the responses of Glossina pallidipes Austen and G.morsitans morsitans Westwood to artificial host odour using an incomplete ring of electrocuting nets. In a plume of synthetic host odour tsetse flew generally upwind, with 50–60% flying within 35^o of due upwind. More than 80% of tsetse flew at < 50 cm above ground level. Upon losing contact with odour they executed a reverse turn within about 2 m, and upon regaining contact they turned upwind. There were no clear differences in the responses of G.m.morsitans and G.pallidipes. Using electrocuting nets lying horizontally on the ground it was found that tsetse landed in the vicinity of the odour source, the propensity to land being greater for G.pallidipes than for G.m, morsitans , greater for immature than mature flies, and greater for males than females.     

Upwind flight of tsetse (Glossina spp.) in response to natural and synthetic host odour in the field

Abstract. In Zimbabwe, studies were made of the flight responses of tsetse ( Glossina spp.) to synthetic and natural ox odour using arrangements of electric nets.Tsetse flying away from a target showed a significant upwind bias when a blend of carbon dioxide (2/1 min), acetone (500 mg/h), octenol (0.4 mg/h), 4-methylphenol (0.8 mg/h) and 3-n-propylphenol (0.1 ma) was dispensed 15 m upwind, with c. 35% flying upwind.Without carbon dioxide this percentage was significantly reduced to 15% which was not significantly different from that with no odour (8%).This pattern was not altered by reducing the doses of acetone, octenol and phenols by 10–100 times, to levels comparable to those produced by an ox.With natural ox odour or a synthetic equivalent of ox odour dispensed from a ventilated pit 8 m upwind of the target, c. 28% flew upwind.This was reduced significantly to 15% if carbon dioxide was removed.In studies using a 17 m line of nets arranged orthogonally across the prevailing wind line, c. 50% of the catch was caught on the downwind side in the absence of odour.This increased significantly to c.60% when acetone, octenol and phenols were dispensed 15 m upwind, with or without carbon dioxide.With a shorter line (9 m) or an incomplete one (16.5 m long with 5 times 1.5 m wide gaps along its length) there was no change in the proportion caught downwind.For all three lines, dispensing odour upwind increased the catch 2–5 times on both the up-and downwind sides of the nets.It is concluded that a stronger upwind response to host odour is elicited when carbon dioxide is present.It is suggested that in nature upwind flight is very imprecisely orientated, with tsetse making flights up and down an odour plume 'searching' for a host.     

The activation of resting tsetse flies (Glossina) in response to visual and olfactory stimuli in the field

ABSTRACT Studies were conducted in Zimbabwe of the responses of Glossina morsitans morsitans Westwood and Glossina pallidipes Austen resting in a refuge to various host stimuli. Tsetse took off in response to 100% ox odour, 0.08% carbon dioxide or a visual stimulus consisting of a 0.75 × 0.75 m black target placed c . 5 m from the refuge moving at 4^o s^-1, but the level of response was low with only 35%, 19% and 29% responding, respectively. Tsetse did not take off in response to any one of 25% ox odour, 0.8% carbon dioxide, acetone (3 μg 1^-1) or octenol (0.03 μg 1^-1). In the absence of any host stimuli, flies emerged from the refuge later on hotter days (35–37^oC) than on cooler days (32–34.5^oC). Male G.pallidipes emerging later in the afternoon contained significantly more haematin than those emerging relatively earlier. There were no significant differences between the responses of G.m. morsitans and G.pallidipes. It is suggested that the initial activation of resting flies is primarily mediated through endogenous, rather than host, stimuli.     

Host odour composition affects host location efficiency of tsetse (Diptera, Glossinidae)

Abstract. Marked Glossina pallidipes Austen were released downwind of an odour source in the field in Zimbabwe and the percentage recaptured at the source on the same day was measured.In the absence of odour, 1.3% of the marked tsetse released from a box or refuge were recaptured, independent of the distance between release point and odour source.The distance was varied from 10 to 100 m.When natural ox odour or a blend of carbon dioxide, acetone, octenol and phenols was dispensed, untransformed recapture percentages of box-released tsetse decreased from 18% for tsetse released at 10 m to 2% for tsetse released at 100 m.Recapture percentages were significantly higher than in the absence of odour at all release distances for ox odour and for release distances up to 75 m downwind for the artificial odour.When a combination of acetone, octenol and phenols or carbon dioxide on its own was dispensed, recapture percentages decreased from 6% for tsetse released at 10 m to 0% for tsetse released at 100 m.With these odours, recapture percentages were higher than in the absence of odour when tsetse were released at 20 m from the source, but were lower than recaptures in the presence of ox odour or the artificial mixture with carbon dioxide.Recapture percentages of flies spontaneously leaving refuges were higher than those of box-released tsetse.Proximity of source had no effect on the recapture percentage of refuge-leaving tsetse and host-location efficiency was close to 100% when host odour was detected at 30 m or less.The results are discussed in relation to the host location strategy of tsetse.     

The host-orientated behaviour of tsetse flies (Glossina): the interaction of visual and olfactory stimuli

ABSTRACT. Studies were made in Zimbabwe of the propensity of Glossina pallidipes Austen and G. morsitans morsitans Westwood to divert from flying upwind in plumes of host odour to various visual features (termed targets). Using various arrangements of electrocuting nets with targets placed downwind of an odour source it was found that 45% diverted to a square target, c. 30% diverted to a black vertical oblong and there was no significant diversion to a bark-coloured vertical oblong that simulated the bole of a tree. The relative propensity of tsetse to divert to variously coloured targets decreased in the order: black = blue > red > yellow; for different shapes it decreased in the order: circle > square > horizontal oblong = vertical oblong. Changes in the composition or concentration of the odour, or loss of contact with it, did not markedly affect the percentage that diverted. Tsetse that diverted to a target and subsequently flew away from it showed an upwind bias in the presence of odour. In the absence of odour there was a slight crosswind bias. If these crosswind fliers then flew into a plume of host odour they turned c. 50 upwind.     

The effect of wind-borne odours on the direction of flight in tsetse flies, Glossina spp.

ABSTRACT. The direction of flight in tsetse flies ( Glossina pallidipes Aust. and G. m. morsitans Westw.) taking off in the presence of certain wind-borne odours showed a significant upwind shift both in the field and in the laboratory. The average angular deviation between the resting orientation and flight direction was not materially affected by odour, but turns were steered in relation to wind direction if odour was present. Upwind flight in an odour plume was regularly preceded by a standing turn, the fly turning partly or completely into the wind before taking off in upwind flight. This suggests that wind direction was assessed, and flight direction determined, before the fly took off.     

Flight responses of tsetse flies (Glossina) to octenol and acetone vapour in a wind-tunnel

Abstract. Female Glossina morsitans morsitans Westwood were video-recorded in a wind-tunnel as they entered, in crosswind flight, a broad plume of either octenol or acetone (two components of ox odour). Both odours produced upwind turning responses (in-flight anemotaxis) to a range of concentrations, with thresholds at around 10^-8mg1^-l for octenol and 10^-6mg1^-1 for acetone. Kinetic responses were unaffected by octenol at low concentrations, but flight speed was significantly reduced and sinuosity (^om^-1) and angular velocity (^os^-1) significantly increased by concentrations at or above those in ox breath; for acetone, these effects were apparent but inconsistently related to concentration. It is concluded that octenol and acetone vapour are used by tsetse flies to locate hosts by upwind anemotaxis, probably combined with kinetic responses. The behavioural basis for the 'repellency' of high octenol concentrations in the field is discussed in the context of the virtual loss of upwind anemotaxis to octenol at the highest concentration tested in the tunnel (30 × ox breath).     

Orientation of tsetse flies to wind, within and outside host odour plumes in the field

Abstract Free-flying wild tsetse flies ( Glossina pallidipes Aust. and G. m. morsitans Westw.) were video recorded in Zimbabwe as they flew within an artificial host odour plume at 3, 7 or 15 m from the source, or in no odour, with and without a 0.75 m² vertical, black visual target present aligned with the wind. With no visual target present, flights in odour were strongly biased upwind, and in the absence of odour strongly biased downwind. With the target present, between 16% and 40% of the upwind approaching flies responded visually as they passed the target, by circling it, in proportion to the proximity of the source (taken to be proportional to the mean odour concentration). Crosswind approaching flies (for whom the target will have been visible for some metres away) circled more frequently (34–56%), but without obvious correlation with the odour concentration. Circling flies also responded orthokinetically, by slowing down as they passed the target. The departure directions relative to the wind of flies leaving the target were significantly affected by the odour concentration. At 3 m they left the target in all directions, except possibly avoiding due upwind. At 7 m they left with an obliquely upwind bias, but at 15 m and also in no odour, they left with a strong crosswind bias.     

Responses of tsetse to ox sebum: a video study in the field

The behaviour of tsetse (mainly Glossina pallidipes Austen) around odour-baited targets, with or without a coating of ox sebum, was recorded in the field using video. The addition of sebum increased the total time a fly was in contact with the target, as well as the time spent flying around and landing on it. When carbon dioxide was released as part of the attractant odour plume, the presence of sebum on the target increased the number of landings made by each fly, but did not significantly affect the duration of each contact. When carbon dioxide was absent from the odour plume, sebum did not affect the number of landings made by flies but the duration of each contact with the target did increase. Evidence for an interactive effect of sebum and carbon dioxide was obtained. In addition, the presence of sebum on the target increased the percentage of landed flies which walked on its surface; such behaviour may represent an 'inspection' of the artificial host. The potential tsetse control application of the current findings are discussed.     

Flight behaviour of tsetse flies in host odour plumes: the initial response to leaving or entering odour

ABSTRACT. Free-flying, wild male and female Glossina pallidipes Aust. and G. m. morsitans Westw. were video-recorded in the field in Zimbabwe as they entered or left the side of a host-odour plume in cross-wind flight, or as they overshot a source of host odour in upwind flight (camera 2.5 m up looking down at a 3 times 2.5 m field of view at ground level). 80% of cross-wind odour leavers turned sharply ( turns 95^o), but without regard to wind direction (overshooters behaved essentially the same except that nearly 100% turned). Many fewer flies entering a plume cross wind turned ( c . 60%), and when they did they made much smaller turns ( 58^o); these turns were, however, significantly biassed upwind ( c . 70%). All three classes of fly had similar groundspeeds ( 5.5–6.5 m s^_1) and angular velocities ( 350–400^o s^-1). Clear evidence was obtained of in-flight sensitivity to wind direction: significantly more flies entering odour turned upwind than downwind, and odour losers turning upwind made significantly larger turns than average. The main basis for the different sizes of turn was the different durations of the turning flight, rather than changes in angular velocity or speed. No evidence was found of flies landing after losing contact with odour.     

Visually-guided, upwind turning behaviour of free-flying tsetse flies in odour-laden wind: a wind-tunnel study

Abstract. To test the hypothesis that tsetse flies use visual input from the apparent movement of the ground to assess wind direction while in flight, Glossina morsitans morsitans Westwood females were video- recorded in a wind-tunnel as they entered, in cross-wind flight, a broad plume of simulated host odour (C0₂ at c. 0.05%). The tunnel (2.3 times 1.2 m wide) generated winds up to 0.25 m s^-1 and had a strongly patterned floor that could be moved upwind or downwind to increase or decrease the visual input due to wind drift. Flight tracks were analysed for speed, direction relative to the wind, and angle of turn. Mean groundspeeds were c. 1.8 m s^-1. In control measurements in still air (with or without odour) flies turned 50:50 'upwind': 'downwind'. With a 0.25 m s^-1 odour-perme- ated wind, 79% turned upwind, and c. 70% left view flying upwind. When the floor was moved at 0.25 m s^-1 upwind (to mimic the visual input from the ground due to a 0.5 m s_^-1 wind), the strength of this response increased. If instead the floor was moved downwind, faster than the wind speed (to mimic the visual input due to a wind from the opposite direction), 59% turned downwind and c. 70% left view flying downwind, and thus away from the source (though progressing 'upwind' in terms of the visual input from apparent ground pattern movement). Upwind turns were on average significantly larger than downwind turns. It is concluded that tsetse navigate up host odour plumes in flight by responding to the visual flow fields due to their movement over the ground (optomotor anemotaxis), even in weak winds blowing at a fraction of their groundspeed.     

Electroantennogram responses of tsetse flies (Glossina pallidipes) to host odours in an open field and riverine woodland

The present study was initiated to gain insight into the way in which tsetse flies ( Glossina spp.) sense odours at different locations in odour plumes in both an open field and a wooded area.
We recorded the antennal responses (EAGs) from stationary living female G. pallidipes 15 m upwind and at various (60, 40, 20, 10, 5 and 1 m) distances downwind from a synthetic host odour source (containing 1-octen-3-ol, acetone and two phenols), in the natural habitat of the fly (Zimbabwe) using a portable electrophysiological device. Experiments were performed in a flat open area (an airstrip) and in riverine woodland. Differences between responses in different environments were determined by comparing various parameters of the EAGs (intermittency, frequency, amplitude, duration and rate of depolarization).
We found that a fly senses odours as puffs that, further downwind, contain less odour and pass less frequently. In an open field downwind from the source, tsetse perceive more olfactory information than upwind for only 10–20 m, whereas in woodland, olfactory responses remain higher and more frequent than upwind up to at least 60 m. In an open field, olfactory information rapidly increases when approaching the odour source from 20 m and in woodland from 5 m onwards.
It is proposed that averaging odour information over time may be of minor importance in long-range location of odour sources. The results suggest that tsetse may smell odour-baited targets from at least 60 m downwind and that the number of flies responding to and being caught by these baits may be higher in woodland than in an open field.     

Tsetse and other biting fly responses to Nzi traps baited with octenol, phenols and acetone

Octenol (1-octen-3-ol), acetone, 4-methylphenol, 3-n-propylphenol, and other potential attractants (human urine, stable fly faeces), as well as guiacol, creosol (potential repellents), were tested as baits for biting flies in North America using standard phthalogen blue IF3GM cotton Nzi traps, or similar commercial polyester traps. Baits were tested during the summers of 2001-04 at a residence in Canada and during January-August 2001 at a dairy in the U.S.A. Behaviour in the presence of octenol was also studied by intercepting flies approaching a trap through the use of transparent adhesive film. Analogous bait and/or trap comparisons were conducted in natural settings in June 1996 in Kenya and in September-December 1997 in Ethiopia. In Canada, catches of five of six common tabanids (Tabanus similis Macquart, Tabanus quinquevittatus Wiedemann, Hybomitra lasiophthalma [Macquart], Chrysops univittatus Macquart, Chrysops aberrans Philip) and the stable fly Stomoxys calcitrans L. were increased significantly by 1.2-2.1 times with octenol (1.5 mg/h). Catches of T. quinquevittatus and S. calcitrans were 3.5-3.6 times higher on a sticky enclosure surrounding a trap baited with octenol. No other baits or bait combinations had an effect on trap catches in North America. In Ethiopia, standard Nzi traps baited with a combination of acetone, octenol and cattle urine caught 1.8-9.9 times as many Stomoxys as similarly baited epsilon, pyramidal, NG2G, S3, biconical and canopy traps, in order of decreasing catch. When baits were compared, catches in Nzi traps of six stable fly species, including S. calcitrans, were not affected by octenol (released at approximately 1 mg/h), or cattle urine (140 mg/h), used alone or in combination with acetone (890 mg/h). Acetone alone, however, significantly increased the catches of common Stomoxys such as Stomoxys niger niger Macquart, Stomoxys taeniatus Bigot, and S. calcitrans by 2.4, 1.6 and 1.9 times, respectively. Catches of Glossina pallidipes Austen were increased significantly in traps baited with acetone, urine or octenol, or any combination, relative to those in unbaited traps (1.4-3.6x). Catches of Glossina morsitans submorsitans Newstead were increased significantly by 1.5-1.7 times, but only when baits were used individually. Unlike other studies with East African tsetse, catches of both tsetse species with the complete bait combination (acetone, urine and octenol) did not differ from those in unbaited traps. Experiments with an incomplete ring of electric nets surrounding a Nzi trap, and a new approach using a sticky enclosure made from transparent adhesive film, revealed diverse responses to artificial objects and baits among biting flies. In Kenya, daily trap efficiency estimates for traps baited with either carbon dioxide (6 L/min) or a combination of acetone, cattle urine and octenol were 21-27% for G. pallidipes, 7-36% for Glossina longipennis Corti, 27-33% for S. n. niger, and 19-33% for Stomoxys niger bilineatus Grünberg, assuming 100% electrocution efficiency. Actual trap efficiencies may have been lower, given observed outside : inside electric net catch ratios of 0.6 : 1.6. Observed ratios averaged 54% of expected values, with 10 of 15 possible ratios less than the minimum possible value of 1.0.     

Dose responses of tsetse flies (Glossina) to carbon dioxide, acetone and octenol in the field

ABSTRACT Studies were conducted in Zimbabwe of the catch of Glossina pallidipes Austen from an electric net plus target baited with mixtures of acetone plus carbon dioxide or 1-octen-3-ol (octenol) plus carbon dioxide. For acetone dispensed alone at 5–50, 000 mg h^-1, ten-fold increments in the dose increased the catch 1.7 times. For carbon dioxide dispensed alone, dose increments from 12 to 1201 h^-1 doubled the catch, but the catch was not further increased by dispensing carbon dioxide at 600–1200 1 h^-1. For mixtures of these two odours, ten-fold increments in the dose of carbon dioxide between 12 and 12, 0001 h^-1 increased the catch c . 2.5 times if acetone was also dispensed at >50 mg h^-1; changes in the dose of acetone between 50 and 50 000 mg h^-1 did not affect the catch. The addition of octenol (0.05 mg h^-1) to carbon dioxide (12–12001 h^-1) doubled the catch. Ten-fold increments in the dose of octenol between 0.05 and 5 mg h^-1 did not increase the catch significantly and the catch was independent of changes in the dose of carbon dioxide between 120 and 12001 h^-1. The behavioural basis of the dose-response curves was investigated using an incomplete ring of electric nets to assess the flight orientation of tsetse in different odours. Upwind flight was not elicited by acetone or octenol alone, or by carbon dioxide unless it was at very high doses, however, mixtures of carbon dioxide with acetone or octenol elicited upwind flight. It is suggested that the attractiveness of mixtures of acetone and carbon dioxide is a function of the region of overlap of these two odours at above threshold concentration. Acetone and octenol on their own appear to increase the responsiveness of flies to visual cues.     

'Anemotactic' flight paths of tsetse flies in relation to host odour: a preliminary video study in nature of the response to loss of odour

ABSTRACT. Free-flying, wild Glossina pallidipes Aust. and G. morsitans Westw. were video-recorded in the field in Zimbabwe as they flew out of air permeated with host odour (camera 2.5 m up, looking down at the ground). Analysis of the flight tracks supports the proposal of Bursell (1984) that tsetse flies attracted to an invisible source of host odour respond weakly if at all to wind direction while in flight: on losing contact with the odour the flies made a sharp turn that was uncorrelated with wind direction. The size of the turn varied considerably, with a marked discontinuity in the log-survivorship curve at 120° (a fly which had turned through at least 120° was 5 times as likely to stop the turn as a fly which had turned <120°). Over half the flies made turns of >90° (and <2 m diameter) within the 2×2.5 m field of view of the camera. It is suggested that these turns initially served to arrest the upwind progress of the fly, with the size of the turn determining the degree to which the fly backtracked towards where it last detected odour or continues cross-wind. Mean flight speed was c. 5 ms^-1 (min. 2.5, max. probably 7ms^-1).     

The effect of wind speed on the flight responses of tsetse flies to CO2: a wind-tunnel study

Abstract. Female Glossina morsitans morsitans Westwood were video-recorded in a wind-tunnel as they entered, in cross-wind flight, a broad plume of CO₂ (a component of host odour). At a wind speed that corresponds with peak catches in the field (c. 0.6 ms-¹) odour produced both significant upwind turning responses (in-flight anemotaxis) and kinetic responses (reduced flight speed and increased sinuosity (m-¹). At a wind speed of c. 0.2 ms-¹ flies displayed anemotactic, but not kinetic, responses to odour. At very low wind speeds (0.1ms-¹) neither upwind turning responses nor kinetic responses to odour were detected. The results are discussed with regard to current theory of host-location by tsetse.     

Observations on the orientation of tsetse flies (Glossina pallidipes) to wind-borne odours

ABSTRACT. Observations of the upwind flight of Glossina pallidipes Austen near a source of host odour show that in the absence of a visual target the insects tend to overshoot the odour source in fast, low flight. There is no sign of the crosswind 'casting' flight which characterizes the behaviour of moths under similar circumstances, except that a 180 turn is executed to bring the tsetse flies back to the vicinity of the odour source in downwind flight. This may be followed by a second overshoot and another 180 turn before the insects alight within a metre or so of the source. The results indicate that the orientation of tsetse flies to host odour may involve a step-wise approach to the odour source, providing an opportunity for assessment of wind direction when the insects are at rest between successive bursts of flight.     

The effect of host odour on the landing responses of tsetse flies (Glossina morsitans morsitans) in a wind tunnel with and without visual targets

Abstract The effect of artificial host odour on the landing responses of males of Glossina m.morsitans West, and on their reaction to visual targets has been investigated in a wind tunnel. Landing was induced in flies that traversed steep odour gradients as they flew upwind and downwind across the edge of an odour plume, irrespective of whether visual targets were present or not; the landing response could be elicited over a wide range of odourconcentrations. When targets were present such odour gradients also tended to increase the proportion of landing flies which alighted on or near the targets; and the bigger the target, or the hungrier the flies, the greater was the propensity for target landing. In air which was more uniformly permeated with odour, the propensity to land on targets was increased only at high odour concentration.     

Interspecific transfer of bacterial endosymbionts between tsetse fly species: infection establishment and effect on host fitness

Tsetse flies (Glossina spp.) can harbor up to three distinct species of endosymbiotic bacteria that exhibit unique modes of transmission and evolutionary histories with their host. Two mutualist enterics, Wigglesworthia and Sodalis, are transmitted maternally to tsetse flies' intrauterine larvae. The third symbiont, from the genus Wolbachia, parasitizes developing oocytes. In this study, we determined that Sodalis isolates from several tsetse fly species are virtually identical based on a phylogenetic analysis of their ftsZ gene sequences. Furthermore, restriction fragment-length polymorphism analysis revealed little variation in the genomes of Sodalis isolates from tsetse fly species within different subgenera (Glossina fuscipes fuscipes and Glossina morsitans morsitans). We also examined the impact on host fitness of transinfecting G. fuscipes fuscipes and G. morsitans morsitans flies with reciprocal Sodalis strains. Tsetse flies cleared of their native Sodalis symbionts were successfully repopulated with the Sodalis species isolated from a different tsetse fly species. These transinfected flies effectively transmitted the novel symbionts to their offspring and experienced no detrimental fitness effects compared to their wild-type counterparts, as measured by longevity and fecundity. Quantitative PCR analysis revealed that transinfected flies maintained their Sodalis populations at densities comparable to those in flies harboring native symbionts. Our ability to transinfect tsetse flies is indicative of Sodalis ' recent evolutionary history with its tsetse fly host and demonstrates that this procedure may be used as a means of streamlining future paratransgenesis experiments.     

Univariate analysis of tsetse habitat in the common fly belt of Southern Africa using climate and remotely sensed vegetation data

Abstract Tsetse are vectors of trypanosomes that cause diseases both in humans and livestock. Traditional tsetse surveys, using sampling methods such as Epsilon traps and black screen fly rounds, are often logistically difficult, costly and time-consuming. The distribution of tsetse, as revealed by such survey methods, is strongly influenced by environmental conditions, such as climate and vegetation cover, which may be readily mapped using satellite data. These data may be used to make predictions of the probable distribution of tsetse in unsurveyed areas by determining the environmental characteristics of areas of tsetse presence and absence in surveyed areas. The same methods may also be used to characterize differences between tsetse species and subspecies. In this paper we analyse the distribution of Glossina morsitans centralis, Glossina morsitans morsitans and Glossina pallidipes in southern Africa with respect to single environmental variables. For G.m.centralis the best predictions were made using the average NDVI (75% correct predictions; range > 0.37) and the average of the maximum temperature (70% correct predictions; 27.0–29.2°C). For G.m.morsitans the best prediction was given by the maximum of the minimum temperature (84% correct predictions; range > 18.8°C), and for G.pallidipes , also by the maximum of the minimum temperature (86% correct predictions; range > 19.6 °C). The following paper compares a range of multivariate techniques for making predictions about the distribution of these species in the same region.