Characterization of the sexual responses of male tsetse flies, Glossina morsitans morsitans, to pheromone-baited decoy 'females' in the field

ABSTRACT. The sexual behaviour of male Glossina morsitans morsitans Westwood (Diptera: Glossinidae) towards decoys dosed with sex phero-mone (15,19,23-trimethylheptatriacontane) is described quantitatively from field observations. The flies responded with a standard behavioural pattern after landing. This involved: first, the perception of pheromone; second, orientation on the decoy and genitalia engagement; and third, a lengthy quiescence corresponding to natural copulation. The probability of a fly leaving a decoy decreased during the sequence and was least in the final 'copulatory' phase, when it remained constant under constant conditions. The initial, rapid rates at which flies left decoys were affected mostly by pheromone dose: the lower the dose, the higher the rate. The final, slow rate of leaving was unaffected by pheromone dose, being most affected by environmental stimuli, especially interference from other flies: at high fly densities, final rates of leaving were high. The results are discussed in the context of improving the potential for using pheromone and chemo-sterilant-dosed decoys for tsetse autosterilization.     

Comparison of the infection rate of tsetse, Glossina morsitans morsitans, fed in vitro or in vivo

Studies were made of infection rates of trypanosomes in the tsetse fly Glossina morsitans morsitans Westwood (Diptera: Glossinidae) when maintained in vivo (rabbits) or in vitro on high quality, gamma-irradiated, sterile defibrinated bovine blood, obtained from the Entomology Unit of the International Atomic Energy Agency (IAEA). For both Trypanosoma congolense Broden and T. b. brucei Plimmer & Bradford, in vitro maintenance significantly reduced the proportion of flies that developed mature metacyclic trypanosome infections.     

Activity patterns in pregnant tsetse flies, Glossina morsitans

ABSTRACT. Actograph measurements of the spontaneous flight activity of isolated female Glossina morsitans morsitans Westw. were made through their first two pregnancy cycles in constant conditions under LD 12:12. Activity fell from a mean of c. 20 units/day to near zero during the 24 h preceding the day on which larvipositionoccurred. Activity on the day of larviposition rose significantly, to c. 35 units/day, then fell back to the baseline 20 units on subsequent days. The circadian pattern of activity was changed markedly by larviposition: 48 h before it, activity in the morning peak of the usual V-shaped diel pattern was twice normal; over 70% of the flies then fed, and the evening arm of their V disappeared; 24 h before larviposition, most flies were totally inactive, the few that did fly performing only 1 or 2 units/day, and only in the first and last hours of the photophase; on the day of larviposition, the morning peak of the V was reduced by c. 70%, and activity peaked instead during the post-noon 2–3 h, before and during the act of parturition. Almost all larvipositions occurred between noon and lights-out. It is suggested that the c. 48 h of pre-larviposition quiescence may play a role in the fact that pregnant G. morsitans females often survive non-residual aerosol insecticide campaigns.     

Genetics of hybridization of Glossina swynnertoni with Glossina morsitans morsitans and Glossina morsitans centralis

Abstract Reciprocal crosses were performed with Glossina swynnertoni and Glossina morsitans morsitans and with G. swynnertoni and Glossina morsitans centralis , using strains that carried marker genes in all three linkage groups. Glossina swynnertoni males can inseminate, but not fertilize, G.m.morsitans; all other crosses produced some fertile females. Hybridization did not cause sex ratio distortion among F_{ flies. Most F and backcross females were fertile, but all F, males were sterile. Sterility among backcross males was also high (99% in BXj, 85% in Bx₂, and about 50% in Bx₃ to Bx₅). Chromosome transmission by hybrid females usually conformed to Mendelian expectations, but genetic recombination was lower than observed in G.m.morsitans. The reduction in fertility among backcross females was not associated with heterozygosity in any linkage group. Sterility among hybrid and backcross males was associated with heterozygosity of sex chromosomes and probably autosomes. The results support the systematic placement of G.swynnertoni closer to G.mxentralis than to G.m.morsitans.     

Lipophorin from the tsetse fly, Glossina morsitans morsitans.

1. Lipophorin was isolated from the haemolymph of adult tsetse fly, Glossina morsitans morsitans, by ultracentrifugation in a potassium bromide density gradient. 2. The tsetse fly lipophorin (Mr congruent to 600,000) has a density of congruent to 1.11 g/ml and consists of two apoproteins, apolipophorin-I (apoLp-I, Mr congruent to 250,000) and apolipophorin-II (apoLp-II, Mr congruent to 80,000), both of which are glycosylated as shown by staining with periodate-Schiff reagent. The protein complex is composed of 49% protein and 51% lipids. 3. The finding of lipophorin in tsetse fly haemolymph suggests that, although these flies primarily utilize proline for their energy needs, there is an active transport mechanism for the supply of lipid requirements.     

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.     

Neuroendocrine stimulation of uterine gland protein synthesis in the tsetse fly, Glossina morsitans

ABSTRACT. The uterine gland of the tsetse fly Glossina morsitans morsitans Westw. synthesizes a secretion which nourishes the developing larva in the uterus. Aqueous extracts of the brain have been shown to stimulate the synthesis of the protein and amino acid components of this secretion from L- [U-¹⁴C]leucine by uterine gland tubules in vivo and in vitro. A linear dose response relationship was demonstrated in vitro with extract concentrations ranging from 1 × 10^-4 to 1 × 10^-2 brains μl^-1. The maximum response, a > 300% increase in the rate of protein and amino acid synthesis, was achieved with as little as 1 × 10^-2 brains μl^-1 The concentration of active factor(s) in the brain declined during a single interlarval period coincident with the period of release of secretion associated with larval growth. The stimulatory activity in brain extracts was destroyed by proteolytic enzymes indicating that it is probably a protein or peptide. Results suggest that the active factor(s) is a hormone responsible for the stimulation of uterine gland protein synthesis essential for larval nutrition.     

Persistent courtship reduces male and female longevity in captive tsetse flies Glossina morsitans morsitans Westwood (Diptera: Glossinidae)

Where males can increase their mating success by harassing femalesuntil they accept copulation, harassing tactics can be expectedto evolve to a point where they have costs to the longevityof both sexes. By experimentally manipulating the sex ratioin captive groups of tsetse flies Glossina morsitans morsitans,we demonstrated that the longevity of females declines wheresex ratios are biased toward males, while the longevity of malesdeclines where the sex ratio is biased toward females. Neitherirradiation of males nor prevention of copulation by blockingor damaging the external male genitalia increased the longevityof females caged with them, suggesting that female longevitywas reduced by the physical aspects of male harassment ratherthan by components of the ejaculate     

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.     

Lectin and peritrophic membrane development in the gut of Glossina m.morsitans and a discussion of their role in protecting the fly against trypanosome infection

Newly emerged Glossina m.morsitans Westwood tsetse flies lack a peritrophic membrane which develops to fully line the midgut after c. 80-90 h. Midgut lectins are mainly associated with the peritrophic membrane. Lectin levels in the blood-free gut of adult flies rise slowly up to 8 days and then rapidly to at least 14 days post-eclosion (when the last of our recordings was made). Despite starving flies for 4 days prior to the agglutination assay, gut lectin levels in older flies are 100-200 times more than those in newly ecloded flies. This is inconsistent with the idea that there is a simple relationship between lectins and the protection of tsetse flies against trypanosome infection. Various theories put forward to account for age-dependent variation in the ability of tsetse to become infected with trypanosomes are discussed in the light of these findings.     

Genetic differentiation of some Glossina morsitans morsitans populations

To study the population structure of Glossina morsitans morsitans Westwood (Diptera: Glossinidae), polymerase chain reaction (PCR) and singlestrand conformational polymorphism (SSCP) methods were used to estimate mitochondrial DNA diversity at four loci in six natural populations from Zambia, Zimbabwe and Mozambique, and in two laboratory cultures. The Zambian and Zimbabwean samples were from a single fly belt. Four alleles were recorded at 12S and 16S1, and five alleles at 16S2 and COI. Nucleotide sequencing confirmed their singularities. Chi-square contingency tests showed that allele frequencies differed significantly among populations. Mean allele diversities in populations averaged over loci varied from 0.14 to 0.61. Little loss in haplotype diversity was detected in the laboratory cultures thereby indicating little inbreeding. Wright's fixation index F(ST) in the natural populations was 0.088+/-0.016, the correlation of haplotypes within populations relative to correlations in the total. A function of its inverse allows an estimate of the mean equivalent number of females exchanged per population per generation, 5.2. No correlation was detected between pairwise genetic distance measures and geographical distances. Drift explains the high degree of differentiation.     

Responses of individual antennal olfactory cells of tsetse flies (Glossina m. morsitans) to phenols from cattle urine

Abstract. Action potentials from olfactory cells in antennae (funiculi) of living tsetse flies, Glossina morsitans morsitans Westwood, were recorded using a surface-contact technique. Stimuli were the vapours of the seven alkylphenols identified in cattle urine: phenol, 3-methyl-, 3-ethyl-, 3- n -propyl-, 4-methyl-, 4-ethyl-, and 4- n -propylphenol. In addition, responses to the vapours of 1-octen-3-ol, acetone and dichloromethane were recorded. The phenol-sensitive cells could be grouped into four subclasses. Subclass, 1, 2 and 3 cells responded to the phenols only, cells of subclass 1 and 2 being activated by these substances, those of subclass 3 inhibited. Cells of subclass 4 were activated to a similar degree by all phenols and by one or more of the other chemicals tested. Subclass 1 cells were strongly activated by the 3-alkylphenols, whereas subclass 2 cells were most sensitive to 4-methylphenol. Subclass 3 cells were most strongly inhibited by phenol, and 3- and 4-methylphenol. The results suggest that though individual phenols may be attractive to G. m. morsitans , preference for certain blends of phenols may exist; for example, blends composed of moderate doses of 4-methylphenol and 3-methyl-, 3-ethyl- or 3- n -propylphenol.     

Spectral sensitivity and flicker fusion frequencies of the compound eye of salmon and wild-type tsetse flies, Glossina morsitans

ABSTRACT. The spectral sensitivity and flicker fusion frequency (FFF) of wild-type and salmon Glossina morsitans morsitans Westwood (Diptera, Glossinidae) were compared electroretinographically (ERG). Spectral sensitivity curves were similar in shape for dark-adapted wild-type and salmon flies, but salmon flies were over 100 times as sensitive as wild-type flies over much of their sensitivity range. Estimation of the spectral absorption curve (from the differences in ERG sensitivities) for the pigment absent from (or present in low concentration in) the salmon eye suggests that the pigment is an ommochrome. FFF at threshold light intensities was similar in wild-type and salmon flies, but at higher light intensity (1.3 °W/cm²) the FFF of salmon flies increased c. 200–300%, due to the capacity of the salmon eye to adapt rapidly to the flicker stimulus. Body weight had little effect upon spectral sensitivity and FFF. Wild-type males were more sensitive to yellow-green light and had higher FFF than did wild-type females. Salmon males and females did not differ in spectral sensitivity, but females had higher FFF (when tested with 520-nm light) than did males. Old wild-type females did not differ from young females in either spectral sensitivity or FFF. However, old salmon females were more sensitive but had lower FFF than young salmon females. Food deprivation reduced spectral sensitivity and FFF in wild-type males but not in salmon males. Irradiation (10.5 krad) reduced spectral sensitivity ( c. 75–375%) and FFF ( c. 30%) in wild-type males. The greatly increased spectral sensitivity and FFF in salmon flies indicate that these flies may behave differently from wild-type flies in the field. Differences in the way spectral sensitivity and flicker discrimination are affected by dark and light adaptation, and by such factors as age and sex, indicate that these measurements are of two independent phenomena.     

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

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

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.     

Determining the age of adult male and female Glossina morsitans morsitans using a new technique

Abstract. 1. Pteridines accumulated linearly with age up to at least 63 days in male and 140 days in female Glossina morsitans morsitans Westwood kept under laboratory conditions. Time is the major factor influencing total pteridine accumulation levels with the accumulation rate being modulated by temperature.
2. It is suggested that pteridine accumulation may be used to determine the age of male and female tsetse flies in the field.     

Ultrastructural studies of certain aspects of the development of Trypanosoma congolense in Glossina morsitans morsitans

The course of Trypanosoma congolense infections in Glossina morsitans morsitans was followed by electron-microscopic examination of ultrathin sections of the guts and proboscises of infected flies. Guts dissected from flies 7 days after infection with culture procyclic forms of T. congolense had heavy trypanosome infections in the midgut involving both the endo- and ectoperitrophic spaces. Trypanosomes were also seen in the process of penetrating the fully formed peritrophic membrane in the central region of the midgut. By post infection day 21, trypanosomes had reached the proboscis of the fly and were found as clumps of epimastigote forms attached to the labrum by hemidesmosomes between their flagella and the chitinous lining of the food canal. Desmosome connections were observed between the flagella of adjacent epimastigotes. Flies examined at postinfection days 28 and 42 had, in addition to the attached forms in the labrum, free forms in the hypopharynx.     

The roles of vision and olfaction in mate location by males of the tsetse fly Glossina morsitans morsitans

The roles of visual and/or olfactory stimuli in eliciting mating responses from male Glossina morsitans morsitans Westwood were examined, using a system for automatically recording the number and duration of mating strikes made towards decoys, under controlled conditions. The results confirm that there is no olfactory component of the female sex recognition pheromone sensed by the male antennae, and the attraction of males to females appears to be visual. The absence of male-male mating strikes was the result of the absence of female sex-pheromone, rather than the presence of a repellent mating deterrent in the male cuticle. Experiments with coloured, artificial, sex-pheromone-dosed, cotton decoys showed that colour had only weak effects on attractiveness and number of encounters with decoys, and that no colour caused significant enhancement of mating responses over those shown to decoy females.