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.     

Effect of host odours on trap catch composition of Glossina pallidipes in Kenya

The effect of odour attractants on the composition of samples of Glossina pallidipes Austen was investigated by comparing the age and nutritional status of flies caught in unbaited biconical traps with those caught in traps baited with cow urine and acetone. For both male and female flies, baited traps caught more flies with significantly higher fat content than did unbaited traps. Thus the samples from baited traps were more representative of the population as a whole: males showed a fuller range of the fat/haematin conditions known to occur in the field and proportionately more females were in later stages of the pregnancy cycle, than from unbaited traps.     

Monitoring tsetse fly populations.:II. The effect of climate on trap catches of Glossina pallidipes

In Part I it was shown that the sampling distribution of trap catches of tsetse flies, Glossina pallidipes Austen, at Nguruman, Kenya, using unbaited biconical traps follows a Poisson distribution. In this paper we examine the effect of humidity and temperature on day-to-day and seasonal variations in the trap catches. It is shown that the seasonal variation is significantly correlated with maximum daily temperature, the catches increasing with temperature when the maximum temperature is below 34 degrees C and decreasing with temperature when it is above 34 degrees C. The correlation between trap catches and relative humidity is not as good as the correlation with the maximum temperature, and the two together do not improve the fit to the trap catches. The day-to-day variation is significantly greater than the intrinsic variation due to the stochastic nature of the sampling process and for some traps it is correlated with temperature and humidity. An autoregressive model gives a half-life for the decay of departures from the mean of about 1 day and it is suggested that this indicates the movement of flies in response to animal movement or to climatic factors other than temperature or humidity. After removing the temperature dependent part of the seasonal variation and the autoregressive component of the data, the male and female catches are still significantly correlated.     

Monitoring tsetse fly populations. I. The intrinsic variability of trap catches of Glossina pallidipes at Nguruman, Kenya

During 1986 the tsetse fly Glossina pallidipes Austen was monitored daily at Nguruman, southwestern Kenya, using three unbaited biconical traps. This was done to investigate the nature and causes of daily variation in trap catches. The variability of the observed catches was compared to a model which includes the trapping probability and the stochastic variation in the sex-ratio. By comparing the catches of male and female flies we are able to establish the sampling distribution of the trap catches. In addition to seasonal changes in the trap catches, day-to-day variations are observed and these are considered greater than the variation arising from the stochastic nature of the sampling process. Recommendations are made in relation to sampling tsetse fly populations.     

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.     

A trial to control the tsetse fly,Glossina morsitans centralis,with low densities of odour-baited targets in west Zambia

Abstract. A large-scale trial investigated the possibility of eradicating G.m.centralis from a traditional cattle rearing area using odour-baited targets at a reduced overall target density from 4 to 0.5-2.3 per km², thus cutting down initial material costs by about 50%. Only the periphery of what was thought to be prime tsetse habitat (dense woodland) was treated with targets. These were all black or blue/black cloth (1.8 × 1m), sprayed with deltamethrin suspension concentrate and baited with butanone and/or acetone (40-130mg/h) and l-octen-3-ol (0.5mg/h). Although fly catches from traps and flyrounds initially dropped by approximately 3% per day and trypanosomiasis cases declined by 99% within a year, eradication was not achieved, so that more targets were deployed at a later stage. Although initially cheaper, the option of using reduced target densities proved financially unattractive because of prolonged periods of target maintenance prior to eradication. Revised strategies for tsetse control with odour-baited, insecticide-impregnated targets in west Zambia are presented.     

Effects of colours and synthetic odours on the attraction of Glossina pallidipes and G. morsitans morsitans to traps and screens

ABSTRACT. Glossina morsitans morsitans Westw. and G. pallidipes Aust. flying around and landing on coloured screens and traps were sampled using electrocuting nets. External colour affected both attractiveness and efficiency of traps, such that yellow and green traps were unattractive and inefficient; black and red, attractive and inefficient; white, moderately attractive, and very efficient; and blue traps, attractive and efficient. The order of attractiveness of coloured screens was similar to that for traps. Landing responses were generally strongest on black surfaces, and weakest on white, but the results for blue were variable. Carbon dioxide and acetone odours improved trap catches and also eliminated most catch differences between traps differently coloured on their outer surfaces. The relative performances of traps coloured differently on inside surfaces only were not affected significantly by these odours, and in all cases black or red target areas inside the trap were required for optimum trap performance. When acetone and 1-octen-3-ol odours were used, catches were improved but the relative performance of differently coloured traps and screens was not changed. There were no obvious species differences in colour responses although numbers of G. morsitans were too low for statistical comparisons.     

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.     

Reproductive abnormalities in tsetse flies in Zimbabwe

Between November 1988 and July 1995 five technicians carried out ovarian dissections on 16,013 Glossina morsitans morsitans Westwood and 123,848 G. pallidipes Austen captured at Rekomitjie Research Station, Zambezi Valley, Zimbabwe. The ovarian age and uterine content were recorded, as were the lengths (l₁, l₂ and l_u) of the largest and second largest oocyte, and of any uterine inclusion. Major abnormalities and abnormal spermathecal contents were found in <0.1% of all flies dissected. Apparent abortions rates varied significantly between dissectors and occurred at frequencies of 0.8–4.5% in G. m. morsitans and 0.3–2.8% in G. pallidipes. The lowest estimates give the best picture of the field situation. Abortion rates were higher in flies caught on electric nets than in trapped flies where the rate was only 0.15%, indicating that reproductive losses are negligible for most of the year at Rekomitjie. The rates did, however, increase to >2% when mean temperatures exceeded 27°C and flies were captured in artificial refuges. There was little effect of ovarian age on the abortion rate, but the frequency of empty uteri declined markedly with age – with a suggestion, however, that it might increase again in the oldest flies. A knowledge of the rates of reproductive loss is important for the construction of realistic models of the dynamics of tsetse populations.     

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.     

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.

---

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.

     

Responses of tsetse flies (Glossina spp.) to compounds on the skin surface of an ox: a laboratory study

The behaviour of male Glossina morsitans morsitans Westwood and Glossina pallidipes Austen (Diptera: Glossinidae) alighting on targets with or without ox sebum was compared. The presence of ox sebum did not increase significantly the number of flies alighting on the target in either species. However, after contact with the sebum coated target, both species showed an increase in flight activity, and G. m.morsitans showed a greater tendency to return to the target. This behaviour resulted in a number of short flights which may reflect the search for a feeding site on a host. The duration of each visit to the target was significantly reduced when sebum was present for G. m. morsitans but not for G. pallidipes. This is explained by documented differences in the resting behaviour of the two species which shows that G. m. morsitans normally rests for longer periods on the surface of an untreated black target than does G. pallidipes. Other experiments showed that the presence of sebum elicited a probing response in G. m. morsitans and G. pallidipes. The results are discussed with reference to the possible use of host sebum to improve trap catches in the field.     

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.

     

The feeding behaviour, activity and trappability of wild female Glossina pallidipe in relation to their pregnancy cycle

Female Glossina pallidipes Austen trapped with baited NG2B traps were subjected both to detailed ovarian dissection and to nutritional analysis. Using a calibration curve derived from dissected wild-caught, laboratory-held flies, the field females were assigned by discriminant analysis to each day of the pregnancy cycle. Field females were most available to NG2B traps while carrying the first instar larva. The nutritional characteristics of trapped field females over the pregnancy cycle lead to the following main conclusions. (i) Fat levels increase most rapidly during the egg in utero stage, while CRDW increases significantly only during the larval stages, culminating in a 4 mg increase during the last day of the third larval instar. (ii) The haematin content of the flies indicates that females feed at approximately 3-day intervals and may feed on any day of the pregnancy cycle. (iii) The estimated time of feeding during the day corresponds with the observed time of peak activity, both of which are earlier in the day later in the pregnancy cycle. (iv) The rate of fat usage reveals significantly greater flight activity on day 5 of the cycle than on other days, agreeing with the high trappability on this day, and overall females appear to use fat at twice the rate of males. Reproductive data provide a much more accurate picture of the relative sampling efficiency than do nutritional data, although the latter reveal the general trends correctly.     

Towards a general rule for estimating the stage of pregnancy in field-caught tsetse flies

Abstract. Ovarian dissections were performed on the tsetse flies Glossina morsitans morsitans Westwood and G.pallidipes Austen of known ages, maintained in the laboratory or on an island in Lake Kariba, Zimbabwe. The lengths ( l ₂ and l _u) of the second largest oocyte and of the larva in utero were found to increase approximately exponentially during pregnancy. The length ( l ₁) of the largest oocyte increased exponentially for about the first 80% of pregnancy. Linear relationships between the log values of l ₁, l ₂ and l _u in field-caught flies, of unknown chronological age, are consistent with the idea that growth patterns are similar in laboratory, island and open field situations. The egg phase takes up c. 45% of pregnancy in both species, regardless of season and the absolute duration of pregnancy. The changes in the log values of l ₁, l ₂ and l _u, over the ranges within which they change linearly, can be used to assign flies to their stage of pregnancy. When applied to field data the rule showed that G.pallidipes caught in odour-baited traps, and on a mobile electric net, exhibited major activity peaks shortly before and after parturition. Flies from the trap (but not the net) showed a smaller peak of activity near the middle of pregnancy. The egg and the three larval phases in utero take up c. 45%, 25%, 20% and 10% of pregnancy respectively.     

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.     

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.     

Genetic variation in Glossina brevipalpis, G.longipennis and G.pallidipes, and the phenetic relationships of Glossina species

Glossina brevipalpis Newstead, G.longipennis Corti, and G.pallidipes Austen maintained at ILRAD, Nairobi, Kenya, were examined for genetic variation of fourteen enzyme loci, using polyacrylamide gel electrophoresis. G.brevipalpis had six polymorphic loci, an average of 1.46 effective alleles per locus and a mean heterozygosity per locus of 20.0 +/- 7.1%. The figures for the same parameters in G.longipennis were 3, 1.16 and 8.2 +/- 4.9%, and for G.pallidipes the figures were 7, 1.40 and 22.3 +/- 6.3%. Seven rare alleles were lost from the G.brevipalpis colony during a 1-year period, but no statistically significant changes were observed in the genetics of the colony during this period. Using allele frequency data for ten of the enzymes studied, and frequencies for these enzymes in other taxa, a phenogram was constructed that indicated that the subgenus Austenina (i.e. the fusca group) is the oldest of the three subgenera within the genus Glossina, and that the subgenus Glossina s.str. (i.e. the morsitans group) may be paraphyletic.     

Analysis of the mating activity of male tsetse flies Glossina m. morsitans and G. pallidipes in the laboratory

ABSTRACT. The responses of male Glossina morsitans morsitans West-wood and Glossina pallidipes Austen to freeze-killed females were examined in the laboratory. Analyses were performed using a specially designed, automated, computer-based, recording system. G. pallidipes were more active than G. m. morsitans , interacting with the female decoys twice as often. Interactions with the decoys divided broadly into short-stay (<60 s) and long-stay, full copulatory attempts. For G. m. morsitans 90% of interactions resulted in full copulatory attempts, the mean duration of which was >1 h. For G. pallidipes only 40% of interactions resulted in full copulatory attempts, the mean duration of which was 35 min. The initiation of interactions showed a clear V-shaped activity pattern in G. m. morsitans but in G. pallidipes only a morning peak was observed. In neither species was there a tendency for full copulatory responses to be initiated in any specific period of the diurnal activity pattern. The results indicate that the two species have very different mating systems, and represent an initial step in the quantification of these differences.     

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.