Nutritional levels of female tsetse Glossina pallidipes from artificial refuges

Female tsetse flies, Glossina pallidipes Austen, caught in artificial refuges were subjected to ovarian dissection and analysed for levels of fat, residual dry weight (RDW) and haematin. There were rather small proportions of flies in ovarian categories 0 and 1, in part due to large losses in the immature and teneral stages at the hottest time of year. The distribution of the female catch among pregnancy days was close to uniform. The wet and dry weights (WW and DW) and RDW of eggs, larvae and pupae increased by 0.821, 0.303 and 0.204 mg respectively, with each mm3 increase in volume. Water accounted for 71.7% of the fat-free WW, and fat for 32.7% of the DW. Between birth and ovulation, fat increased from 2 to 4 mg and RDW from 7 to 11 mg; thoracic RDW increased by 2.5mg and changed little thereafter. Fat levels increased 3.5mg by day 6 of pregnancy, but only 0.5 mg thereafter. Over the same periods RDW corrected to zero haematin (CRDW) increased by 1 and 8 mg respectively. Full-term fat and CRDW levels were 8.2 and 19.4 mg respectively. Cumulative haematin frequencies formed a smooth curve with a slope that increased continuously. The raw data were well fitted by a model where feeding rates increased exponentially and capture probability was independent of haematin content. The mean feeding interval was 60 h; feeding probabilities of >0.9/day were only found in flies that had failed to feed for>72 h. In early pregnancy, fat levels declined with haematin for flies that had fed>36 h previously; by days 5-7 fat levels were maintained at a constant high level for 60 h post-feeding. Fat-haematin graphs for female tsetse cannot be used to estimate rates of fat utilization. Traps sample tsetse with below-average fat and RDW in early and late pregnancy respectively. Refuge samples are less biased than those from traps; they give a better picture of the dynamics of pregnancy in normal flies and facilitate the explanation of existing anomalies.     

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.     

Lifetime changes in the nutritional characteristics of female tsetse Glossina pallidipes caught in odour-baited traps

Female Glossina pallidipes Austen were captured in odour-baited traps at Rekomitjie Research Station, Zambezi Valley, Zimbabwe during February 1994; 2890 were dissected and assigned to their ovarian age category and day of pregnancy using the lengths of the oocytes and uterine content. For 1838 of these flies, the nutritional state of the mother and her uterine content were estimated separately. It was thereby possible to see how, during pregnancy, the females acquired fat and residual dry weight (RDW) and transfered them to the larva. Newly emerged flies contained 1 mg fat and 6 mg RDW, of which 4 mg was in the thorax (TRDW). Fat hardly increased by the first ovulation; RDW increased by 2.5 mg and 1.5 mg of this increase was in TRDW. Mean haematin levels increased from 2 to 8 microg during each pregnancy. Fat increased from 1.2 mg to 4.5-5 mg by day 7 and was then rapidly transferred to the larva. RDW increased by only 1.8 mg by day 7, but larval RDW increased thereafter by > 6 mg. Amino acids from late-pregnancy bloodmeals are incorporated directly, in the uterine gland, into 'milk' that is taken up rapidly by the larva. Capture probability was highest on day 1 of pregnancy, when nutritional levels were lowest, with lesser peaks on days 5 and 8 when the fly was nourishing a rapidly developing larva. On day 1, the peak of the logarithm of the haematin distribution corresponded to flies estimated to have fed approximately 75 h previously; by day 8 it had shifted to approximately 60 h post-feeding. A model in which feeding rates and capture probabilities increased exponentially with time since feeding accounted for 97% of the variance in log haematin frequencies. On 4/9 days of pregnancy there was no significant decline in fat with haematin content during the lipolytic phase. The rate of decline is not a satisfactory estimate of the rate of fat usage. Flies in this study had longer wings and higher TRDW than those from refuges in an earlier study, but had lower levels of fat and haematin.     

Performance of the tsetse fly Glossina pallidipes reared under simple laboratory conditions

A simple grass-thatched hut was used to rear Glossina pallidipes Austen. Constructed to allow free flow of air, the temperature and humidity inside correspond to that of the outside environment. Pupae used to start the colony were obtained from pregnant females from Lambwe Valley, Western Kenya. Nine day-old females were paired with 12 day-old males for 7 days after which they were separated.Colony performance was evaluated on the basis of female survival, number and weight of puparia produced, and mortality rates. The emergence rate of puparia weighing more than 30 mg was above 80%. Data on the performance of the colony between June 1985 and June 1986 are presented. We suggest that the simple nature of the rearing facility and its direct contact with the outside environment have facilitated the colonization of the flies.

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Résumé Une cage très simple, conçue pour assurer l'arrivé d'air, et le maintien de l'humidité et de loa température extérieure, a été employée pour conserver et élever G. pallidipes. Les pupes utilisées pour constituer la souche provenaient de Lambwe valley (ouest du Kénya). Des couples de femelles vieilles de 9 jours et de mâles de 12 jours ont été consitués, les sexes ont ensiute été séparés. La performance de la souche a été estimée en fonction de la survie des femelles et du nombre et du poids des pupes produites, et des taux de mortalité. Plus de 80% des pupes ont pesé 30 mg et plus, et le taux d'émergence a été de 80%. Les résultats fournis concernent les performances observées entre juin 1985 et juin 1986. Nous estimons que la simplicité du dispositif d'élevage et le contact direct avec les conditions écologiques externes ont facilité l'éstablissement de la souche.

     

Evaluation of insecticide-treated cattle as a barrier to re-invasion of tsetse to cleared areas in northeastern Zimbabwe

A field trial in Zimbabwe investigated the efficacy of insecticide-treated cattle as a barrier to prevent the re-invasion of tsetse, Glossina morsitans and G. pallidipes (Diptera: Glossinidae), into cleared areas. The original tsetse barrier consisted of insecticide-treated odour-baited targets, at an operational density of four to five targets per km2, supported by insecticide-treatments of cattle with either deltamethrin dip (Decatix, Coopers) at two-weekly intervals, or deltamethrin pouron (Spoton, Coopers) at monthly intervals, in a band approximately 20 km wide from the re-invasion front. Tsetse catch, and trypanosomiasis incidence in nine sentinel herds was recorded for 7-8 months, respectively, before the targets were removed, leaving only the insecticide treatment of the local cattle to stem the re-invasion of tsetse. After the removal of the target barrier, the tsetse readily invaded the trial area and the incidence of trypanosomiasis in sentinel herds increased, while their PCVs decreased. After seven months without the targets in place, trypanosomiasis prevalence in the local stock had reached alarmingly high levels; the trial was terminated prematurely and the target barrier re-deployed. Immediately after the re-deployment of the target barrier, the tsetse catch in the trial area reverted to acceptable levels along the re-invasion front, and trypanosomiasis incidence in the sentinel cattle decreased. It is concluded that, under the conditions of the field trial, the insecticidal treatment of local cattle did not in itself form an effective barrier to tsetse re-invasion. By contrast, the target barrier performed as was predicted by mathematical and experimental analysis, and readily cleared the tsetse infestation and reduced trypanosomosis incidence in the trial area.     

Ovulation, insemination and mating in the tsetse fly Glossina pallidipes

ABSTRACT. Analyses of the ovulation, insemination and mating of female Glossina pallidipes Austen, were carried out in the field and laboratory. In unmated females, ovulation was delayed until at least the fifteenth day of adult life. The predictable pattern of growth and development of eggs was used to age nulliparous females to the nearest day in the field. Inseminated females first appeared at 5–6 days old and most females became inseminated at 8–10 days of age. There were no significant differences in the age at insemination in Zimbabwe or Kenya. Ovulation had occurred in the field by 10–11 days old. Mating therefore occurs at or just before ovulation. Females were able to mate at up to 12 days old and still produce a viable larva from their first egg. In laboratory flies originating from Uganda, factors which reduced and minimized the effects of disturbance, maximized insemination rate, so that under the best conditions the age at mating was identical to that found in the field. Nevertheless, the insemination rate of Fl generation Zimbabwe flies reared from wild-caught females was negligible. The differences in the laboratory mating behaviour of the Zimbabwe flies and the flies from the long-established colony of Uganda origin, are considered to be due to wild G. pallidipes being easily disturbed under laboratory conditions, whereas colony breeding rapidly selects for passivity. The concept of gross differences in G. pallidipes mating behaviour between geographic areas in thus rejected.     

Size and mortality rates of Glossina pallidipes in the semi-arid zone of southwestern Kenya

Seasonal changes in the mean size of tsetse, Glossina pallidipes Austen, as indicated by wing vein length, were monitored during 1983-86 at Nguruman, southwestern Kenya. Changes in size of nulliparous females and wing fray category 1 males were shown to be correlated with the relative humidity 2 months before they were captured. Soil temperature when flies were in the pupal stage had much less effect. Size dependent mortality was demonstrated, with the mean size of flies emerging from pupae significantly less than that of field-caught flies. This mortality must occur at emergence, since there was no evidence of size-dependent mortality once the flies became available to the trap. Size was correlated with density-independent mortality acting on the parent population 2 months previously. It might therefore be possible to use size as an index of the intensity of such mortality. This could be useful when assessing the level of additional mortality required to suppress tsetse populations.     

Copulatory behaviour of the tsetse flies Glossina morsitans and G.austeni

ABSTRACT. The copulatory behaviour of Glossina morsitans morsitans West. and G.austeni Newst. was analysed by filming. After a male and female engaged genitalia, the male performed a repertoire of five actions for 3–4 min: (1) 'rubbing' his metathoracic, tibiotarsal joint against the region of genital contact; (2) 'stroking' or hitting the female's head and thorax with his meso- and metathoracic legs; (3) 'wing flick' by moving his mesothoracic legs in a rowing motion whilst at the same time vibrating the wings as in normal flight; (4) 'wing vibration' with the wings vibrated in the closed position; (5) 'wings out' in which the wings are moved out to the flying position without any observable vibration. Each action was repeated many times, to give variable individual sequences, but declined in frequency exponentially over the first 3–4 min in copulo. The two species differed in the frequency of acts. Shortly before separation, a few hours later, actions 1 and 2 reappeared. Receptive females exhibited little overt behaviour except in the maintenance of a passive stance. Refractory females rejected a mating attempt by flexing the abdomen ventrally, vibrating their wings in the closed position, and pushing the male with meso- and metathoracic legs. The significance and possible functions of male behaviour are discussed in relation to mating in Glossina and other Diptera.     

Influence of temperature changes on larviposition rhythm in the tsetse fly, Glossina morsitans

ABSTRACT. The hourly and quarter-hourly distribution of larval deposition by female Glossina morsitans Westw. was studied at various temperatures. At constant temperature and humidity, peak larviposition occurred 8–9 h after lights on. The effect of confining the flies to daily (LD 12:12 h) regimes of at least twenty-five consecutive cycles that included a sudden increase in temperature of c. 6°C for 2 h was a significant reduction in depositions during each period of increased temperature, and a significant increase before the heat impulse. When the temperature was allowed to fall rapidly, larviposition rate doubled in the following 15 min.     

Abundance and distribution of the tsetse flies, Glossina austeni and G. brevipalpis, in different habitats in South Africa

The distribution and abundance of Glossina austeni Newstead and Glossina brevipalpis Newstead (Diptera: Glossinidae) were studied in the three main vegetation types in Zululand, KwaZulu-Natal, South Africa. During a period of 12 months, a trap transect consisting of 38 H-traps traversing the three vegetation types was monitored. The Index of Apparent Abundance (IAA) for G. brevipalpis was high in indigenous forest and open grassland but lower in exotic plantations. Glossina austeni, on the other hand, was captured mainly in or adjacent to indigenous forest. The seasonal trend in the IAA did not differ between vegetation types. The findings on the distribution of G. brevipalpis are in contrast with the historic records. Historically, this species was considered to be restricted to areas with a dense overhead canopy and high relative humidity. The repercussions of these findings for the epidemiology of livestock trypanosomiasis and the control of tsetse in Zululand are discussed.     

Reproductive rates of tsetse flies in the field in Zimbabwe

Abstract. Tsetse flies Glossina morsitans morsitans Westwood and G. pallidipes Austen were marked and released within 12 h of emergence at Rekomitjie Research Station, Zambezi Valley, Zimbabwe, and on Redcliff Island, Lake Kariba. Ovarian dissections were performed on recaptured flies and on wild collected samples. At Rekomitjie >90% of female G. m. morsitans were inseminated by age 4 days and G. pallidipes by 7 days. For both species at both sites the length of the largest oocyte, for flies in ovarian category zero, increased approximately linearly for about the first 6 days and was ovulated at c. 6–8 days. The largest oocyte grew significantly more slowly in later cycles. For G. m. morsitans , but not for G. pallidipes , the rate increased with temperature; the rates were always higher than observed in the laboratory. At Rekomitjie, for both species and at a mean screen temperature of 22C, the first larva was produced at c. 18 days and subsequent larvae at 11–day intervals; the intervals decreased with temperature by c. 0.5 days/C. On Redcliff Island the intervals for both species were 2 days shorter than at Rekomitjie at any given screen temperature and were sometimes as short as 7 days. The length of the larva in utero increased exponentially during pregnancy.     

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.     

Development of a low-cost tsetse trap and odour baits for Glossina pallidipes and G.longipennis in Kenya

Experiments were carried out to improve the NG2B tsetse trap (Brightwell et al., 1987), baited with acetone and cow urine, for use by rural communities to control G.pallidipes Austen and G.longipennis Corti. Modifications included a lower dose rate of acetone, a new cage design and raising the trap about 15-20 cm. Research on different trap cone materials showed that the degree of light transmission of the netting, rather than its colour, was the crucial factor affecting the catch of G.pallidipes. Adding an additional metre of blue cloth to one side of the trap increased catches of females of both species by about 60%. Traps baited with synthetic phenols yielded similar numbers of G.pallidipes and significantly more G.longipennis than those baited with natural cow urine. The latter difference was not apparent when octenol was also used, so cow urine was retained as one of the odour baits in preference to the imported phenols. Although octenol increased catches of G.pallidipes by only about 30%, catches of G.longipennis were increased 2-4-fold, making it a very useful attractant for the latter species. The cost of the trap/odour-bait system was estimated to be US$8.5 per unit per annum. The economics of this method of tsetse control are discussed.     

Reciprocal regulation of fat content and flight activity in male tsetse flies (Glossina palpalis)

Abstract. To disentangle cause and effect in previously observed relationships between fat content and flight activity in male tsetse ( Glossina spp.), three groups of flies were fed at different intervals to raise their fat content to different levels before their flight activity was recorded. The greater the mean daily blood intake, the higher the fat content and the greater the subsequent spontaneous flight activity, thereby using up almost all of the fat reserves before the next blood meal. It is proposed that although male flies would benefit from maximum food intake to permit maximum flight associated with mate-seeking, they do not in fact feed as often as possible either in the field or the laboratory. This is explicable if energy acquisition is constrained by an additional mortality risk associated with feeding.     

Microgeographical breeding structure of the tsetse fly, Glossina pallidipes in south-western Kenya

The origins of extant Glossina pallidipes Austen (Diptera: Glossinidae) populations in the ecologically well-studied Lambwe and Nguruman valleys in Kenya are controversial because populations have recovered after seemingly effective attempts to achieve high levels of control. The microgeographical breeding structure of the tsetse fly, G. pallidipes, was investigated by analysing spatial and temporal variation at eight microsatellite loci to test hypotheses about endemism and immigration. Samples were obtained at seasonal intervals from trap sites separated by 200 m to 14 km and arranged into blocks. G. pallidipes populations nearest to Lambwe and Nguruman also were sampled. Spatial analysis indicated that genetic differentiation by genetic drift was much less among trapping sites within Lambwe and Nguruman (F(ST) < or = 0.049) than between them (F(ST) = 0.232). F(ST) between Serengeti and Nguruman was 0.16 and F(ST) between Kodera Forest and Lambwe was 0.15. The genetic variance in G. pallidipes explained by dry and wet seasons (0.33%) was about one-fifth the variance among collection dates (1.6%), thereby indicating reasonable temporal stability of genetic variation. Gene frequencies in Kodera and Serengeti differed greatly from Lambwe and Nguruman, thereby falsifying the hypothesis that Lambwe and Nguruman were repopulated by immigrants. Harmonic mean effective (= breeding) population sizes were 180 in Lambwe and 551 in Nguruman. The genetic data suggest that G. pallidipes in Lambwe and Nguruman have been endemic for long intervals.     

Selective use of odour-baited, insecticide-treated targets to control tsetse flies Glossina austeni and G. brevipalpis in South Africa

The effectiveness of odour-baited targets treated with 0.8% deltamethrin in controlling Glossina austeni Newstead and G. brevipalpis Newstead (Diptera: Glossinidae) was evaluated in Zululand, South Africa. Targets were initially deployed in the three habitat types (grassland, woodland and forest) of two adjacent areas at a density of four targets per km(2). One area functioned as the treatment block (c. 35 km(2)) and included the focus of the target deployment, and the second area functioned as a barrier block (c. 40 km(2)) against tsetse fly re-invasion from the untreated area to the south. After 8 months, targets were removed from open grassland in both areas and target density in wooded habitats and sand forest was increased to eight per km(2). Twelve months later, all targets were removed from the barrier block and used to increase target density in the wooded and sand forest habitats of the treatment block to 12 per km(2). This target density was maintained for 14 months. In the treatment area, a 99% reduction in G. austeni females occurred after 13 months at a target density of eight per km(2) in wooded habitat; this was maintained for 22 months. Reduction in G. brevipalpis was less marked. The relatively poor reduction in G. brevipalpis is attributed to the high mobility of this species and its distribution throughout less wooded and more open habitats.     

Bloodmeal evacuation from the midgut of Glossina pallidipes in relation to tsetse foraging activity and trappabiIity

Abstract.

• 1 Foraging for bloodmeals is the most frequently recurrent and probably the most targetable of all activities that render tsetse vulnerable to interception with static trapping devices.
• 2 Surgical monitoring of the rnidgut every 24 h during three successive days of food deprivation, showed that a full bloodmeal, irrespective of its size or source, vacated the rnidgut of both sexes of Glossina pallidipes Austen in eight progressive stages.
• 3 Probing responsiveness in both sexes increased exponentially during the first four stages of their midgut evacuation, reaching a peak between stages 3 and 5. Thereafter it decreased, also exponentially.
• 4 Most G.pallidipes caught by NG2G traps baited with cow urine and acetone had midguts in the last three stages (6–8) of bloodmeal evacuation. The same was true of the majority of those that failed to feed on a calf shortly after entrapment.
• 5 The implications of the foregoing for tsetse foraging activity and trappability as well as for the potency of cattle urine and acetone as odour-bait for tsetse are discussed.

     

Development of a system for sterilizing tsetse flies, Glossina spp., in the Held

Various autosterilizing systems were evaluated on natural populations of Glossina pallidipes Austen and G.morsitans morsitans Westwood in Zimbabwe. These involved a clear plastic tower of two or three chambers mounted at the cage position on a trap. Inside one chamber flies could be exposed to the vapour phase of the chemosterilant bisazir, P,P-bis (1-aziridinyl)-N-methylphosphinothioic amide. The system was designed to encourage flies to enter the sterilizing chamber, delaying their exit for sufficient time to expose them to a sterilizing dose. This was accomplished in the most effective system by restriction of the exit to one small hole (6 mm diameter) at the roof-side junction. The number of flies remaining in the chamber declined exponentially. The rate of exit was directly proportional to the density of flies in the sterilizing chamber and to the number of exit holes. The probability of a fly being in this chamber for at least 1 or 7 min (the times taken for female and male G.m.morsitans respectively to receive an ED50) was 0.84 and 0.67 respectively with one fly present. With sixteen flies, these probabilities were 0.76 and 0.18 respectively. Results suggest that it may be possible to develop a cheap, safe and efficient autosterilizer for use on tsetse traps.     

A model for the relationship between wing fray and chronological and ovarian ages in tsetse (Glossina spp)

Age-dependent mortality changes in haematophagous insects are difficult to measure but are important determinants of population dynamics and vectorial capacity. A Markov process was used to model age-dependent changes in wing fray in tsetse (Glossina spp), calibrated using published mark–recapture data for male G. m. morsitans in Tanzania. The model was applied to female G. m. morsitans, captured in Zimbabwe using a vehicle-mounted electric net and subjected to ovarian dissection and wing fray analysis. Rates of fray increased significantly with age in males but not females, where the rate was constant for ovarian categories 0–3. A jump in mean fray between ovarian categories 3 and 4 + 4n is consistent with the latter category including flies that have ovulated 4, 8, 12, 16 times and so on. The magnitude of the jump could, theoretically, facilitate improved mortality estimates. In practice, although knowledge of fly mortality was required for modelling wing fray, mortality estimates derived from ovarian dissection data are independent of patterns and rates of change in wing fray. Significantly better fits to ovarian age data resulted when age-specific mortality was modelled as the sum of two exponentials, with high mortality in young and old flies, than when mortality was constant at 2.3% per day.