The effect of starvation on the susceptibility of teneral and non-teneral tsetse flies to trypanosome infection

Transmission of vector-borne diseases depends largely on the ability of the insect vector to become infected with the parasite. In tsetse flies, newly emerged or teneral flies are considered the most likely to develop a mature, infective trypanosome infection. This was confirmed during experimental infections where laboratory-reared Glossina morsitans morsitans Westwood (Diptera: Glossinidae) were infected with Trypanosoma congolense or T. brucei brucei. The ability of mature adult tsetse flies to become infected with trypanosomes was significantly lower than that of newly emerged flies for both parasites. However, the nutritional status of the tsetse at the time of the infective bloodmeal affected its ability to acquire either a T. congolense or T. b. brucei infection. Indeed, an extreme period of starvation (3-4 days for teneral flies, 7 days for adult flies) lowers the developmental barrier for a trypanosome infection, especially at the midgut level of the tsetse fly. Adult G. m. morsitans became at least as susceptible as newly emerged flies to infection with T. congolense. Moreover, the susceptibility of adult flies, starved for 7 days, to an infection with T. b. brucei was also significantly increased, but only at the level of maturation of an established midgut infection to a salivary gland infection. The outcome of these experimental infections clearly suggests that, under natural conditions, nutritional stress in adult tsetse flies could contribute substantially to the epidemiology of tsetse-transmitted trypanosomiasis.     

Inhibition of the DNA amplification of trypanosomes present in tsetse flies midguts: implications for the identification of trypanosome species in wild tsetse flies

The present study was carried out in order to investigate if there was really a failure of PCR in identifying parasitologically positive tsetse flies in the field. Tsetse flies (Glossina palpalis gambiensis and Glossina morsitans morsitans) were therefore experimentally infected with two different species of Trypanosoma (Trypanosoma brucei gambiense or Trypanosoma congolense). A total of 152 tsetse flies were dissected, and organs of each fly (midgut, proboscis or salivary glands) were examined. The positive organs were then analysed using PCR. Results showed that, regardless of the trypanosome species, PCR failed to amplify 40% of the parasitologically positive midguts. This failure, which does not occur with diluted samples, is likely to be caused by an inhibition of the amplification reaction. This finding has important implications for the detection and the identification of trypanosome species in wild tsetse flies.     

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.     

Olfactory responses of tsetse flies to phenols from buffalo urine

Abstract A comparison was made of the EAG responses of males and females of Glossina morsitans morsitans Westwood, G.austeni Newstead and G.tachinoides , Westwood to various doses of compounds known to be components of ox and buffalo urine fractions which are attractive to tsetse in the field (phenol, 3- and 4-methylphenol, 3- and 4-ethylphenol, 4-n-propylphenol, dimethylsulfone). All three species did not respond to dimethylsulfone. The overall responses to the phenolic substances were higher in females than in males in G.m.morsitans and higher in males than in females in G.austeni and G.tachinoides. Response spectra of the species for the phenolic substances suggested that G.m.morsitans and G. austeni were most responsive to 3- and 4-methylphenol and 3-ethylphenol, whereas G. tachinoides was most sensitive to 3-ethylphenol and 3-methylphenol, and only moderately sensitive to 4-methylphenol.
Cross-adaptation experiments, in which l-octen-3-ol, acetone, 4-heptanone and 3-nonanone were also included, revealed that all phenolic compounds stimulated one and the same class of receptors, which differed from the class of receptors activated by l-octen-3-ol. The ketones also had their own receptors. Hence, the flies can obtain information about the presence of attractants by at least three different receptor classes. It was concluded that phenol and any individual alkylphenol found in ox and buffalo urine should be attractive to tsetse flies, provided that stimulus intensity is above threshold and not beyond optimum. One class of receptors may respond more strongly in males than in females, whereas another class is more responsive in females than in males. This may result in a change in sex ratios in catches depending on the odour bait used.     

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.     

Effect of host packed cell volume on the bloodmeal size of male tsetse flies, Glossina pallidipes

aematin contents of engorged, male tsetse flies, Glossina pallidipes Austen, were compared with the packed cell volumes of oxen on which they had fed. Haematin contents icnreased with packed cell volume up to packed cell volumes of approximately 30%. Haematin contents appeared to level off or decline with further increase in packed cell volume. These results support a model of blood-feeding in tsetse flies in which the rate of blood consumption decreases as packed cell volume increases, because of increase in blood viscosity, and tsetse are unable to compensate for the decrease in consumption rate by feeding for a longer time. After allowing for the effects of packed cell volume, bloodmeal sizes of tsetse increased with ox body temperature.     

Haemolymph lectin and the maturation of trypanosome infections in tsetse

The tsetse immune system has recently been shown to be involved in trypanosome maturation; lectin secreted in the midgut, normally responsible for preventing the establishment of midgut infections, induces established midgut trypanosomes to mature. We now show that a second lectin, present in tsetse haemolymph, is essential to complete the maturation process. Interactions between tsetse lectins and parasite surface coats probably determine trypanosome transmissibility and may be partly responsible for the distribution of trypanosomiasis in Africa.     

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.     

Reproductive under-performance of tsetse flies in the laboratory, related to feeding frequency

Abstract. The rates of development of the eggs and larvae in utero and the next two developing ovarioles were measured by ovarian dissection on each day of the pregnancy cycle in tsetse, Glossina morsitans, subject to different feeding regimes. Compared with flies fed four times per pregnancy cycle, flies fed three times per cycle showed a lower pupal production rate (70%), the same (zero) adult mortality, a slightly slower growth rate of the larva and second ovariole only from day 8 onwards, but the same growth rate of the first ovariole. Flies fed only twice per pregnancy cycle produced no pupae, suffered 18% adult mortality and showed a significantly slower growth rate of the larva and second ovariole from days 6 and 7 respectively, but still the growth rate of the first ovariole was barely affected. Flies offered food three times or twice per pregnancy cycle engorged fully at every opportunity, but 16.5% of the flies offered food four times per cycle did not feed on every occasion, while 12–22% did not engorge fully on days 3, 5 or 7. In assessing the applicability of these laboratory results to the field situation the following points must be borne in mind: in the laboratory flies take smaller mean blood meals than in the field; during protein production associated with larval growth the proportion of the blood meal lost to transformation and excretory costs is less than during normal lipid metabolism; the balance between the tsetse's known fertility rate and adult and pupal mortality rates reveals that the abortion rate in the field must be extremely low. The high abortion rates usually observed in laboratory colonies, even when flies are offered food daily_l would be quite untenable in the field and indicate that laboratory conditions impose physiological stresses on the flies that are quite different from those in the field. These facts indicate that three field-sized meals may be sufficient to meet the energy demands of normal larval development in the field.     

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.     

Coenzyme A precursors flow from mother to zygote and from microbiome to host

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Odour movement, wind direction, and the problem of host-finding by tsetse flies

ABSTRACT. Movement of host odour was modelled in natural tsetse habitats with smoke and ultra-light 7-cm-long wind vanes; the speed and direction of the air movements were analysed from video recordings thereof. Wind of <1 ms^-1 did not move in straight lines, since large packets of air (>10 m across) often changed direction together. The rate of this change of direction (meander) correlated negatively with windspeed. In open woodland with a shrubby understorey (in which windspeed was reduced by a factor of >5 from that above the canopy, to ax 0.3 m s^-1), this wind meander fell by 2d? s^-1 change of direction for each 0.1 m s^-1 increase in windspeed (r²=0.96). Over open ground without shrub cover, the meander fell by 0.5d? s^-1 per 0.1 m s^-1 increase in windspeed (r²=0.85). In both situations, such meandering virtually ceased in winds of > 1 m s^-1. In woodland, the relationship between the direction of air movement near the surface of bare earth (one potential tsetse landing site) and that c. 0.5 m above ground level (flight height) was often weak (r²=0.2-0.4), but this problem would be reduced if the fly averaged the ground-level wind for at least 30 s. Odour (smoke) travelling from a source 15 m ‘upwind’ over open ground arrived at a notional tsetse fly for 80% of the time from a direction within 10d? of the true source direction. In typical tsetse woodland, however, the ‘odour’ arrived from all directions (including >90d? away from the source), with only a 30% bias towards the true source direction (±10d?). Evidently, tsetse must navigate up odour plumes by means that get round these difficulties-simple, moth-type upwind anemotaxis alone seems unlikely to be adequate.     

Condition and fate of larvae from insecticide-treated female tsetse flies Glossina palpalis

Many larvae from wild-caught female Glossina palpalis palpalis R.-D. treated topically with dieldrin or sumicidin or sumithrin or permethrin failed to pupariate and many larvae from mothers treated with sumicidin or sumithrin or neopynamin formed small or malformed puparia which failed to eclode. Puparia produced soon after maternal handling often did not emerge and many produced by flies starved more than 3 days also did not emerge. Very many puparia from dieldrin-treated mothers failed to emerge and non-emergence was common for puparia from flies treated with endosulfan or sumithrin or deltamethrin. Non-emerging puparia were produced soon after pyrethroid-treatment, probably due to effects on larval integumentand muscles, but were produced later after organochlorine-treatment, due to ingestion of insecticide by larvae while in utero.

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Résumé Les expériences ont porté sur des femelles sauvages de Glossina palpalis palpalis R.D. capturées dans la nature. Les comparaisons ont porté sur l'état et les performances de lots de pupes provenant de mouches non-traitées et de larves mûres traitées localement avec de l'acétone (témoin) ou avec 14 insecticides différents.Toutes les larves témoin se sont nymphosées, mais le taux d'échec augmentait avec la diéldrine, la sumicidine, la sumithrine et la permethrine. Les pupes anormales (petites ou déformées) étaient plus nombreuses dans le lot témoin que dans le lot non-traité, et les fréquences étaient plus élevées que chez le témoin avec la sumidicine, la sumithrine et la néopynamine.Seulement 3 mouches sont issues de 107 pupes anormales; il n'y a eu aucune éclosion pour 28% des pupes du lot témoin contre 15% pour le lot non-traité. Pour le témoin le taux d'émergence était faible dans les pupes formées le premier jour (jour du traitement), par suite de la manipulation maternelle; il n'a pas augmenté jours 2 et 3, mais a diminué par la suite à cause du jeûne in utero. Les doses élevées de tous les insecticides réduisent plus les émergences que les doses faibles. Les organophosphates et le propoxur ne limitent pas les émergences, mais elles sont réduites par l'endosulfane, la sumithrine et la deltaméthrine, et très fortement par la diéldrine. Les pupes stériles ont été surtout formées dès le traitement aux pyréthroïdes (jour 1); et surtout plus tard (jours 2 et 3) lors des traitements aux organochlorés. L'ingestion par la larve in utero de grandes quantités d'insecticides pendant les 3 jours précédant la parturition était la principale cause de mortalité, et aussi de la forte tolérance aux organochlorés des femelles prégnantes. Les effets de pyréthroïdes sur l'activité musculaire réduisent aussi les émergences.Aucune mouche ne s'est alimentée après les traitements.

     

A spatial genetics approach to inform vector control of tsetse flies (Glossina fuscipes fuscipes) in Northern Uganda

Tsetse flies (genus Glossina) are the only vector for the parasitic trypanosomes responsible for sleeping sickness and nagana across sub‐Saharan Africa. In Uganda, the tsetse fly Glossina fuscipes fuscipes is responsible for transmission of the parasite in 90% of sleeping sickness cases, and co‐occurrence of both forms of human‐infective trypanosomes makes vector control a priority. We use population genetic data from 38 samples from northern Uganda in a novel methodological pipeline that integrates genetic data, remotely sensed environmental data, and hundreds of field‐survey observations. This methodological pipeline identifies isolated habitat by first identifying environmental parameters correlated with genetic differentiation, second, predicting spatial connectivity using field‐survey observations and the most predictive environmental parameter(s), and third, overlaying the connectivity surface onto a habitat suitability map. Results from this pipeline indicated that net photosynthesis was the strongest predictor of genetic differentiation in G. f. fuscipes in northern Uganda. The resulting connectivity surface identified a large area of well‐connected habitat in northwestern Uganda, and twenty‐four isolated patches on the northeastern margin of the G. f. fuscipes distribution. We tested this novel methodological pipeline by completing an ad hoc sample and genetic screen of G. f. fuscipes samples from a model‐predicted isolated patch, and evaluated whether the ad hoc sample was in fact as genetically isolated as predicted. Results indicated that genetic isolation of the ad hoc sample was as genetically isolated as predicted, with differentiation well above estimates made in samples from within well‐connected habitat separated by similar geographic distances. This work has important practical implications for the control of tsetse and other disease vectors, because it provides a way to identify isolated populations where it will be safer and easier to implement vector control and that should be prioritized as study sites during the development and improvement of vector control methods.     

Enhancing vector refractoriness to trypanosome infection: achievements,challenges and perspectives

With the absence of effective prophylactic vaccines and drugs against African trypanosomosis, control of this group of zoonotic neglected tropical diseases depends the control of the tsetse fly vector. When applied in an area-wide insect pest management approach, the sterile insect technique (SIT) is effective in eliminating single tsetse species from isolated populations. The need to enhance the effectiveness of SIT led to the concept of investigating tsetse-trypanosome interactions by a consortium of researchers in a five-year (2013–2018) Coordinated Research Project (CRP) organized by the Joint Division of FAO/IAEA. The goal of this CRP was to elucidate tsetse-symbiome-pathogen molecular interactions to improve SIT and SIT-compatible interventions for trypanosomoses control by enhancing vector refractoriness. This would allow extension of SIT into areas with potential disease transmission. This paper highlights the CRP’s major achievements and discusses the science-based perspectives for successful mitigation or eradication of African trypanosomosis.     

Responses of tsetse flies, Glossina morsitans morsitans and Glossina pallidipes, to baits of various size

Recent studies of Palpalis group tsetse [Glossina fuscipes fuscipes (Diptera: Glossinidae) in Kenya] suggest that small (0.25 × 0.25 m) insecticide-treated targets will be more cost-effective than the larger (≥1.0 × 1.0 m) designs currently used to control tsetse. Studies were undertaken in Zimbabwe to assess whether small targets are also more cost-effective for the Morsitans group tsetse, Glossina morsitans morsitans and Glossina pallidipes. Numbers of tsetse contacting targets of 0.25 × 0.25 m or 1.0 × 1.0 m, respectively, were estimated using arrangements of electrocuting grids which killed or stunned tsetse as they contacted the target. Catches of G. pallidipes and G. m. morsitans at small (0.25 × 0.25 m) targets were, respectively, ～1% and ～6% of catches at large (1.0 × 1.0 m) targets. Hence, the tsetse killed per unit area of target was greater for the larger than the smaller target, suggesting that small targets are not cost-effective for use against Morsitans group species. The results suggest that there is a fundamental difference in the host-orientated behaviour of Morsitans and Palpalis group tsetse and that the former are more responsive to host odours, whereas the latter seem highly responsive to visual stimuli.