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.     

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.     

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.     

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.     

The efficacy of various pyrethroid insecticides for use on odour-baited targets to control tsetse

The efficacy of various pyrethroid insecticides for use on odour-baited targets to control tsetse was compared in Zimbabwe. Formulations were applied to cotton cloth and polyester net and, at various intervals, the materials were bioassayed by exposing fed female Glossina pallidipes (Austen) (Diptera: Glossinidae) to cloth for 45 s or by inducing them to collide briefly with net. Trial formulations were compared with deltamethrin suspension concentrate (s.c.), the insecticide currently used in tsetse control operations in Zimbabwe. Applying 0.8% suspension of alphacypermethrin to cloth or net produced high mortalities for 9 months which was similar in performance to 0.4% suspension of deltamethrin s.c. Deltamethrin s.c. and beta-cyfluthrin s.c. applied to cloth as 0.1% suspensions were equally effective, producing high mortalities for 2 months during the wet season, and 0.8% suspension of beta-cyfluthrin was effective for 12 months. Suspensions of 0.1% lambdacyhalothrin capsule suspension or 0.1% lambdacyhalothrin wettable powder were significantly less effective than 0.1% deltamethrin s.c. Chemical analyses showed that increasing the concentration of insecticide applied to material increased the initial amount of insecticide on the material and decreased the subsequent rate of loss; 0.1% suspension of beta-cyfluthrin s.c. applied to cloth produced an initial concentration of approximately 280 mg/m2 which declined by 94% in 12 months whereas 0.8% suspension showed no significant decrease in concentration (mean= 1304 mg/m2) over the same period. For controlling tsetse by means of pyrethroid-treated targets, it is suggested that beta-cyfluthrin s.c. is as effective as deltamethrin s.c. but that alphacypermethrin s.c. should be used at twice the concentration of deltamethrin s.c. to obtain the same performance.     

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.     

Sex pheromone of the tsetse species, Glossina austeni: isolation and identification of natural hydrocarbons, and bioassay of synthesized compounds

Copulatory responses of male Glossina austeni (Newstead) (Diptera: Glossinidae), that were elicited after contact with frozen female tsetse, were not observed after solvent washing of cuticular lipids. Chromatographic analysis of extracts from laboratory-reared and field-collected G. austeni females yielded natural hydrocarbons that were highly stimulatory to males. Most of this activity was produced by compounds in the alkene fraction. Gas chromatograms (GC) contained five natural alkenes; these were separated by preparative GC for bioassays conducted in Tanzania. The two major alkenes were identified using gas chromatography-mass spectrometry (GC-MS) to be 13,17-dimethyltritriacont-1-ene and 13,17-dimethylpentatriacont-1-ene, after the samples had undergone derivatization using dimethyl disulphide and saturation with deuterium. These alkenes and natural alkanes were quantified from G. austeni of both sexes from laboratory and field samples to confirm that their presence was consistent in this species. Trials of synthetic samples resulted in the order of biological activity for the stereoisomers of 13,17-dimethyltritriacont-1-ene as follows: S,R-33:1 > R,S- 33:1 > S,S-33:1 > R,R-33:1. Dose-response data showed an ED(50) at 5 microg per treated, solvent-washed male decoy. Of the four stereoisomers of 13,17-dimethylpentatriacont-1-ene, R,R-35:1 showed the most activity. This is the first report of alkene-induced sexual activity in males of the genus Glossina.     

Responses of Glossina austeni to sticky panels and odours

The responses of male tsetse Glossina austeni Newstead (Diptera: Glossinidae) towards blue and white sticky legged panels, baited with odour attractants, and towards modified panels were studied in the Jozani forest of Unguja Island, Zanzibar. Increasing the height of the body of a standard panel from 30 to 60 or 90 cm, increased the catch two-fold. Increasing the height of the legs (from 15 to 60 or 120 cm) or raising the device more than 5 cm above the ground reduced the catch significantly. The legs of the panels were the preferred landing sites of the flies, irrespective of the height of the body of the panel. Acetone (300 mg/h) combined with cow urine (60-130 mg/h) significantly increased the catches two- to threefold during the rainy season, but not during the dry season. Acetone had no effect during the dry season and its effect during the rainy season was less consistent. There was no effect of octenol (2.5-12.5 mg/h), used alone or in combination with acetone. Likewise, the catch did not increase through the addition of cow sebum, pig urine (60-860 mg/h), pig urine combined with acetone and octenol. The observed seasonal differences in the response of G. austeni towards odours are discussed in relation to host location strategies.     

Tissue distribution and prevalence of Wolbachia infections in tsetse flies, Glossina spp

Tsetse flies Glossina spp. (Diptera: Glossinidae) harbor three different symbiotic microorganisms, one being an intracellular Rickettsia of the genus Wolbachia. This bacterium infects a wide range of arthropods, where it causes a variety of reproductive abnormalities, one of which is termed cytoplasmic incompatibility (CI) that, when expressed, results in embryonic death due to disruptions in fertilization events. We report here that in colonized flies, Wolbachia infections can be detected in 100% of sampled individuals, while infections vary significantly in field populations. Based on Wolbachia Surface Protein (wsp) gene sequence analysis, the infections associated with different fly species are all unique within the A group of the Wolbachia pipientis clade. In addition to being present in germ-line tissues, Wolbachia infections have been found in somatic tissues of several insects. Using a Wolbachia-specific PCR-based assay, the tissue tropism of infections in Glossina morsitans morsitans Westwood, Glossina brevipalpis Newstead and Glossina austeni Newstead were analysed. While infections in G. m. morsitans and G. brevipalpis were limited to reproductive tissues, in G. austeni, Wolbachia could be detected in various somatic tissues.     

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.     

Field responses of Glossina austeni to sticky panels on Unguja Island, Zanzibar

Abstract .Two designs of cross-shaped sticky panels (XT and XLP) were compared with the royal blue–white legpanel (LPBuWh) in the Jozani forest on Unguja Island as trapping devices for male Glossina austeni . Single coloured royal blue (XTBu) and bi-coloured royal blue–white XT (XTBuWh) caught more than twice as many male G. austeni as the LPBuWh, whereas single coloured black XT trapped significantly fewer flies (10%) than the control LPBuWh. XT's in various horizontal and diagonal blue–white configurations likewise trapped more flies than the LPBuWh, except a horizontally striped blue–white XT which trapped fivefold fewer flies than the LPBuWh. Cross-shaped LP in the blue–white (XLPBuWh) and black–white (XLPBlWh) combination scored significantly better than the control LPBuWh. Similar fly numbers were trapped with XTBuWh and XLPBuWh. Long-term trapping data indicated that the XTBu, XTBuWh and XLPBuWh were three- to fourfold more effective in trapping female flies than the LPBuWh.
The landing bias on bi-coloured panels was low in the blue–white but more pronounced in the blue–black and white–black combinations and was affected by the type of sticky panel used. A high proportion (49%) of the flies alighted on the bottom corners of the XTBu panel, but landing positions were more scattered if white was added.
Increasing the width of the XTBu from 70 to 120 cm improved the catch by a factor of two as compared with standard sized XTBu. The effect of doubling the height of the XT on total fly catch was negligible.
At present, it is the XTBu which can be recommended as the best trapping device for male and female G. austeni.     

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.     

A study on the maturation of procyclic Trypanosoma brucei brucei in Glossina morsitans centralis and G. brevipalpis

Abstract. Teneral Glossina morsitans centralis and G. brevipalpis were fed in vitro upon medium containing procyclic Trypanosoma brucei brucei derived from the midguts of G. m. centralis or G. brevipalpis which had immature trypanosome infections. The tsetse were then maintained on rabbits and, on day 31, were dissected to determine the infection rates. In G. m. centralis the midgut and salivary gland infection rates by T. b. brucei were 46.0% and 27.0% with procyclic trypanosomes from G. m. centralis, and 45.4% and 24.7% with procyclic trypanosomes from G. brevipalpis, respectively. In G. brevipalpis the rates were 20.2% and 0.0% with procyclic trypanosomes from G. m. centralis, and 28.0% and 0.0% with procyclic trypanosomes from G. brevipalpis, respectively. Teneral G. m. centralis and G. brevipalpis were also fed similarly upon procyclic T. b. brucei derived from G.m.centralis or G. brevipalpis on day 31 of infection, the former tsetse species had mature infections while the latter were without infections in the salivary glands. In G.m.centralis the infection rates in the midgut and salivary glands were 48.9% and 17.0%, and 38.0% and 17.0% when fed on procyclic trypanosomes from G.m.centralis and G. brevipalpis, respectively. In G. brevipalpis the rates were 21.5% and 0.0%, and 10.7% and 0.0% with procyclic trypanosomes of G.m.centralis and G. brevipalpis origin, respectively. Thus, procyclic T. b. brucei from susceptible G.m.centralis could not complete cyclical development in refractory G. brevipalpis, whereas those from G. brevipalpis developed to metatrypanosomes in the salivary glands of G.m.centralis. Teneral and 15-day-old non-teneral G.m.centralis were fed in vitro upon heparinized goat's blood containing T. b. brucei bloodstream trypomastigotes, or upon medium containing procyclic T. b. brucei derived from G.m.centralis with mature infections. On day 31 their infection rates were determined. The infection rates by T. b. brucei in the midgut and salivary glands of G.m.centralis fed on the infected blood were 70.4% and 40.4% when fed as teneral tsetse, as against 15.3% and 4.0% when fed as non-teneral tsetse. Those tsetse which were fed on the medium containing procyclic trypanosomes showed rates of 50.0% and 25.6%, as against 11.6% and 2.5%, respectively. It would appear, therefore, that maturation of T. b. brucei in tsetse is probably not determined simply by an interaction between lectin and procyclic trypanosomes in the midgut of non-teneral tsetse, but it is the result of a complex interaction between many interrelated physiological factors of both the trypanosome and the tsetse vector.     

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.     

Microsatellite diversities and gene flow in the tsetse fly,Glossina morsitans s.l

Tsetse flies occupy discontinuous habitats and gene flow among them needs to be investigated in anticipation of area-wide control programs. Genetic diversities were estimated at six microsatellite loci in seven Glossina morsitans submorsitans Newstead (Diptera: Glossinidae) populations and five microsatellite loci in six G. m. morsitans Westwood populations. Nei's unbiased diversities were 0.808 and 76 alleles in G. m. submorsitans and 0.727 and 55 alleles in G. m. morsitans. Diversities were less in three laboratory cultures. Matings were random within populations. Populations were highly differentiated genetically. Populations were strongly subdivided, as indicated by fixation indices (F(ST)) of 0.18 in G. m. morsitans and 0.17 in G. m. submorsitans. 35% of the genetic variance in G. m. submorsitans was attributed to differences between populations from The Gambia and Ethiopia. All available genetic evidence suggests that genetic drift is much greater than gene flow among G. morsitans s.l. populations.     

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.     

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.     

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.