New data on parasitization of Glossina palpalis palpalis (Diptera: Glossinidae) by Hexamermis glossinae (Nematoda: Mermithidae) in a forested area of Ivory Coast

Observations on the parasitism of Glossina palpalis palpalis by Hexamermis glossinae were carried out over a period of one year by catching flies at Abengourou, Aboisso and Daloa (forested area of Ivory Coast). No parasite is observed out of 2,168 Glossina palpalis palpalis caught in Abengourou and 9,732 in Aboisso. At Daloa, dissections of 7,341 Glossina reveal 1.75% parasited flies. All the worms were located in the abdominal cavity, loosely intertwined with the internal organ. Males were more infested than females (2.68% versus 1.26%). The parasites were more abundant among the nulliparous (2.30%) than the young parous (1.19%) and than the old parous (0.52%). The majority of infected flies were caught at the beginning of the rainy season (5.17%) and few in the dry season (0.23%). The low parasitic infection rate observed here indicates a minimal effect on the population dynamics of the vector of sleeping sickness in Ivory Coast.     

Mitochondrial diversity analysis of Glossina palpalis gambiensis from Mali and Senegal

West African riverine tsetse populations of Glossina palpalis gambiensis Vanderplank (Diptera: Glossinidae) were investigated for gene flow, inferred from mitochondrial diversity in samples of 69 flies from Senegal and 303 flies from three river drainages in Mali. Four polymorphic mitochondrial loci were scored. Mean haplotype diversities were 0.30 in Mali, 0 in Senegal and 0.18 over both Mali and Senegal. These diversities estimate the probabilities that two randomly chosen tsetse have different haplotypes. Substantial rates of gene flow were detected among flies sampled along tributaries belonging to the river basins of the Senegal, Niger, and Bani in Mali. There was virtually no gene flow between tsetse in Senegal and Mali. No seasonal effects on gene flow were detected. The implications of these preliminary findings for the implementation of area-wide integrated pest management (AW-IPM) programmes against riverine tsetse in West Africa are discussed.     

Spatial distribution and bloodmeal preferences of Glossina palpalis palpalis in the forest focus of Zoukougbeu (Ivory Coast). Epidemiological consequences

In the sleeping sickness focus of Zoukougbeu (C?te d'Ivoire), in the cropping areas which are favourable for disease transmission, more than a quarter of the flies collected were found to have fed on domestic pigs. The sites where Glossina palpalis palpalis was caught fed on these animals were concordant with the sites where the patients were present. These results might indicate that in Zoukougbeu, but perhaps also in other sleeping sickness foci, the pig could play an active role in disease transmission, allowing the parasite to spread widely via the tsetse.     

Effect of the life-span of female Glossina palpalis gambiensis on the weight and size of its progeny

Abstract. Pupae and teneral flies of Glossina palpalis gambiensis originating from three successive reproductive cycles were compared for their size and weight. In general, pupal weight and fly weight increased, whereas fly size, measured as wing vein length, decreased with the number of reproductive cycles. The linear regression observed between weight and wing vein length of the fly demonstrated that, particularly for flies originating from the first and second larvipositions, small changes in wing vein length reflected substantial differences in weight.
The results of these laboratory experiments were compared with some field data on Glossina morsitans from Zambia and related literature. The life span of the female tsetse, affecting the size of her progeny, could clarify partially some of the field observed seasonal changes in size, whereas the correlation between fly size and weight could eventually explain the differential mortality of some size classes of tsetse flies. However, whether these laboratory observations can be extrapolated to the field has still to be confirmed.     

Intraspecific variability in natural populations of Glossina palpalis gambiensis from West Africa, revealed by genetic and morphometric analyses

Glossina palpalis gambiensis Vanderplank (Diptera: Glossinidae) from West Africa (Senegal and Burkina Faso) were analysed for microsatellite DNA polymorphisms and size of the wings. In the overall sample a strong heterozygote deficiency was found at two polymorphic microsatellite loci. It led to a highly significant value of Fis (within-sample heterozygote deficit) in the western zone of Sideradougou area in Burkina Faso. Genetic differentiation was significant on a macrogeographic scale, i.e. between tsetse coming from Senegal and Burkina Faso. Wing measures also differed between these two countries; flies from Senegal appeared to be smaller. Microsatellite loci further allowed differentiation of populations of G. palpalis gambiensis trapped on the same hydrographic network a few kilometres apart. The results are interpreted as indicating that further investigations will allow the study of genetic variability of tsetse flies in relation to the dynamics of transmission of human and animal trypanosomoses.     

Predicted distribution and movement of Glossina palpalis palpalis (Diptera: Glossinidae) in the wet and dry seasons in the Kogo trypanosomiasis focus (Equatorial Guinea).

The aim of this study was to predict the distribution and movement of populations of the tsetse fly, Glossina palpalis palpalis (Diptera: Glossinidae), in the wet and dry seasons and to analyze the impact of the use of mono-pyramidal traps on fly populations in the Kogo focus in 2004 and 2005. Three Glossina species are present in Kogo: Glossina palpalis palpalis, major HAT vector in West-Central Africa, Glossina caliginea, and Glossina tabaniformis. The apparent density (AD) of G. p. palpalis clearly fell from 1.23 tsetse/trap/day in July 2004 to 0.27 in December 2005. A significant reduction in the mean AD for this species was noted between seasons and years. The diversity of Glossina species was relatively low at all the sampling points; G. p. palpalis clearly predominated over the other species and significantly dropped as a consequence of control activities. The predictive models generated for the seasonal AD showed notable differences not only in the density but in the distribution of the G. p. palpalis population between the rainy and dry season. The mono-pyramidal traps have proven to be an effective instrument for reducing the density of the tsetse fly populations, although given that the Kogo trypanosomiasis focus extends from the southern Equatorial Guinea to northern Gabon, interventions need to be planned on a larger scale, involving both countries, to guarantee the long-term success of control.     

Trypanosome characterization by polymerase chain reaction in Glossina palpalis gambiensisand G.tachinoidesfrom Burkina Faso

Abstract. Following the discovery of four cases of African human trypanosomiasis, an entomological survey was conducted along the Mouhoun river in southwest Burkina Faso to collect Glossina palpalis gambiensis and G.tachinoides. Among 226 flies dissected, 4.87% (eleven individuals) were infected in midgut or proboscis, but never in the salivary glands. Polymerase chain reaction analysis was undertaken, and was able to characterize all the proboscis infections, and half of the midgut infections. Only Trypanosoma simiae and T. vivax were found in the organs of infected flies, in single or mixed-species infections. Ten more flies, negative with parasitological examination, were tested with Trypanozoon primers and remained negative. The epidemiological significance of the absence of T.brucei group infections in wild tsetse populations and the presence of T.simiae in G.p.gambiensis are discussed.     

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

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

Identification of Glossina palpalis gambiensis specific salivary antigens: towards the development of a serologic biomarker of human exposure to tsetse flies in West Africa

The saliva of blood sucking arthropods contains a number of pharmacologically active compounds that induce an antibody response in exposed human individuals. The objectives of the present study were (i) to assess the human IgG response directed against salivary antigens of Glossina palpalis gambiensis, the main vector of Trypanosoma brucei gambiense in West Africa, as a biomarker of human–tsetse contacts; and (ii) to identify specific salivary antigens. Immune reactivity of human plasma collected within active human African trypanosomiasis (HAT) foci (coastal Guinea), historical foci where tsetse flies are still present (South-West Burkina Faso) and a tsetse free area (Bobo-Dioulasso, Burkina Faso), was measured by ELISA against whole saliva extracts. In the active HAT foci and areas where tsetse flies were present in high densities, specific IgG responses were significantly higher (p < 0.0001) to those in Bobo-Dioulasso or in Loropeni, where tsetse flies were either absent or only present at low densities. Furthermore, 2D-electrophoresis combined with mass spectrometry enabled to reveal that several antigens were specifically recognized by plasma from exposed individuals. Among them, four salivary proteins were successfully identified (Ada, 5′Nuc, Ag5 and Tsgf1). These results represent a first attempt to identify Glossina salivary proteins or synthetic peptides to develop a standardized and specific biomarker of tsetse exposure in West Africa.     

Glossina morsitans morsitansandGlossina palpalis palpalis:Dosage Compensation Raises Questions about the Milligan Model for Control of Trypanosome Development

Gooding, R. H., and McIntyre, G. S. 1998.Glossina morsitans morsitansandGlossina palpalis palpalis: Dosage compensation raises questions about the Milligan model for control of trypanosome development.Experimental Parasitology90, 244–249. Evidence that dosage compensation occurs in tsetse flies was obtained by comparing the activities of X chromosome-linked enzymes, arginine phosphokinase and glucose-6-phosphate dehydrogenase inGlossina m. morsitansand hexokinase and phosphoglucomutase inGlossina p. palpalis, with the activity of an autosome-linked enzyme, malate dehydrogenase, in each species. The shortcomings of the X chromosome model for the control ofTrypanozoonmaturation in tsetse are discussed in light of these findings and previously published reports on the lack of fitness effects of matureTrypanozooninfections in tsetse and on published results on antitrypanosomal factors in male and female tsetse flies.     

Geographical distribution of Glossina palpalis gambiensis and G.p.palpalis in Liberia

The two subspecies of Glossina palpalis (Robineau-Desvoidy) occurring in Liberia could be reliably separated morphometrically by measuring the width of the terminal dilatations of the male inferior claspers. Subspecies differentiation of female flies was less conclusive. Identification of flies from fifty-four sites revealed that most of Liberia lies in the belt of Glossina palpalis palpalis. However, pure and substantial populations of G.p.gambiensis Vanderplank occur north of 8 degrees 20'N in the north-west of Liberia.     

Molecular phylogenetics of tsetse flies (Diptera: Glossinidae) based on mitochondrial (COI, 16S, ND2) and nuclear ribosomal DNA sequences, with an emphasis on the palpalis group

Relationships of 13 species of the genus Glossina (tsetse flies) were inferred from mitochondrial (cytochrome oxidase 1, NADH dehydrogenase 2 and 16S) and nuclear (internal transcribed spacer 1 of rDNA) sequences. The resulting phylogeny confirms the monophyly of the morphologically defined fusca, morsitans and palpalis subgenera. Genetic distances between palpalis and morsitans subspecies suggest that their status needs revision. In particular, cytochrome oxidase 1 sequences showed large geographical differences within G. palpalis palpalis, suggesting the existence of cryptic species within this subspecies. The morphology of palpalis group female genital plates was examined, and individuals were found varying outside the ranges specified by the standard identification keys, making definitive morphological classification impossible. A diagnostic PCR to distinguish G. palpalis palpalis, G. tachinoides and G. palpalis gambiensis based on length differences of internal transcribed spacer 1 sequences is presented.     

Circadian activity patterns in three species of tsetse fly: Glossina palpalis, austeni and morsitans

ABSTRACT. Observations on the nature and control of spontaneous flight activity in tsetse flies have so far been made only on G. m. morsitans Westw. This paper reports preliminary observations of the same kind on two other species: G. p. palpalis (R.-D.) and G. austeni Newst. The flight pattern of all three species consisted of short bursts of activity (usually lasting <1 min) separated by long intervals of rest, changes in activity level occurring exclusively as changes in the frequency of these flight bursts. In constant conditions in LD 12:12, G. palpalis and G. austeni were almost totally diurnal, with strongly predominant afternoon activity peaks, but a slight tendency to decrease activity around noon as occurs to a much greater extent in G. morsitans. This rhythm persisted in constant darkness (DD), but whereas in most G. morsitans only the morning peak was retained in DD, in G. palpalis and G. austeni it was only the afternoon peak. The loss of the afternoon peak in G. morsitans may be due to the very low activity levels in DD. Food deprivation led to increased activity in all three species, most markedly so in G. palpalis and G. austeni , in which the amount of activity per day increased c. ten-fold over three days' post-emergence starvation.     

Increased expression of unusual EP repeat-containing proteins in the midgut of the tsetse fly (Glossina) after bacterial challenge

Proteins containing a glutamic acid-proline (EP) repeat epitope were immunologically detected in midguts from eight species of Glossina (tsetse flies). The molecular masses of the tsetse EP proteins differed among species groups. The amino acid sequence of one of these proteins, from Glossina palpalis palpalis, was determined and compared to the sequence of a homologue, the tsetse midgut EP protein of Glossina m. morsitans. The extended EP repeat domains comprised between 36% (G. m. morsitans) and 46% (G. p. palpalis) of the amino acid residues, but otherwise the two polypeptide chains shared most of their sequences and predicted functional domains. The levels of expression of tsetse EP protein in adult teneral midguts were markedly higher than in midguts from larvae. The EP protein was detected by immunoblotting in the fat body, proventriculus and midgut, the known major immune tissues of tsetse and is likely secreted as it was also detected in hemolymph. The EP protein was not produced by the bacterial symbionts of tsetse midguts as determined by genome analysis of Wigglesworthia glossinidia and immunoblot analysis of Sodalis glossinidius. Bacterial challenge of G. m. morsitans, by injection of live E. coli, induced augmented expression of the tsetse EP protein. The presence of EP proteins in a wide variety of tsetse, their constitutive expression in adult fat body and midguts and their upregulation after immunogen challenge suggest they play an important role as a component of the immune system in tsetse.     

Gorging response of Glossina palpalis palpalis to ATP analogues

ABSTRACT. The potencies of seventeen analogues of ATP as gorging inducers for Glossina palpalis palpalis were evaluated. The ranking for effective dose that induced half the flies to gorge (ED₅₀) was: A tetra P 5 ATP=2'd ATP ADP=2'd ADP > AMP-PNP > 3'd ATP 2'3'dd ATP > AMP-PCP > adenosine 5' triphosphate 2',3'dialdehyde AMP-CPP >> AMP. Females detect ATP and its analogues better than males. The ED₅₀ of ATP was 5 × 10^-7 M for teneral females and 1.5 × 10^-6 M for males. According to the potency order of the ATP analogues, the G.p.palpalis gustatory receptors recognizing ATP can be classified as P_2y purinoceptors.     

Sodalis glossinidius (Enterobacteriaceae) and Vectorial Competence of Glossina palpalis gambiensis and Glossina morsitans morsitans for Trypanosoma congolense Savannah Type

Sodalis glossinidius is an endosymbiont of Glossina palpalis gambiensis and Glossina morsitans morsitans, the vectors of Trypanosoma congolense. The presence of the symbiont was investigated by PCR in Trypanosoma congolense savannah type-infected and noninfected midguts of both fly species, and into the probosces of flies displaying either mature or immature infection, to investigate possible correlation with the vectorial competence of tsetse flies. Sodalis glossinidius was detected in all midguts, infected or not, from both Glossina species. It was also detected in probosces from Glossina palpalis gambiensis flies displaying mature or immature infection, but never in probosces from Glossina morsitans morsitans. These results suggest that, a) there might be no direct correlation between the presence of Sodalis glossinidius and the vectorial competence of Glossina, and b) the symbiont is probably not involved in Trypanosoma congolense savannah type maturation. It could however participate in the establishment process of the parasite.     

Evidence for a discrete evolutionary lineage within Equatorial Guinea suggests that the tsetse fly Glossina palpalis palpalis exists as a species complex

Tsetse flies of the palpalis group are major vectors of Human African Trypanosomiasis in Africa. Accurate knowledge of species identity is essential for vector control. Here, we combine ribosomal internal transcribed spacer 1 ( ITS1 ), mitochondrial Cytochrome Oxidase 1 ( COI ) and microsatellites to determine the population structure and phylogenetic relations of Glossina p. palpalis in Equatorial Guinea. CO1 sequence data suggest that G. p. palpalis in Equatorial Guinea is a distinct subspecies from previously described G. p. palpalis in West Africa and Democratic Republic of Congo. Glossina p. palpalis in Equatorial Guinea and DRC share a common ancestor which diverged from West African G. p. palpalis around 1.9 Ma. Previous ITS1 length polymorphism data suggested the possible presence of hybrids in Equatorial Guinea. However, ITS1 showed incomplete lineage sorting compared with clearly defined COI groups, and data from 12 unlinked microsatellites provided no evidence of hybridization. Microsatellite data indicated moderate but significant differentiation between the populations analysed (Rio Campo, Mbini and Kogo). Moreover, unlike previous studies of G. p. palpalis , there was no evidence for heterozygote deficiency, presence of migrants or cryptic population structure. Variance effective population size at Rio Campo was estimated at 501–731 assuming eight generations per year. This study of the population genetics of G. p. palpalis in central Africa provides the first estimate of genetic differentiation between geographically separated G. p. palpalis populations.     

Trypanosoma brucei 29-13 strain is inducible in but not permissive for the tsetse fly vector

Using green fluorescent protein as a reporter, we have shown that the strain 29-13 of Trypanosoma brucei, widely used for inducible down-regulation of mRNA, is inducible in, but not permissive for the tsetse flies Glossina palpalis gambiensis and Glossina morsitans morsitans. Within two weeks post-infection, 42% males and females of teneral and non-teneral tsetse flies harboured intestinal infections, yet not a single infection progressed into the salivary glands.     

A new transmission risk index for human African trypanosomiasis and its application in the identification of sites of high transmission of sleeping sickness in the Fontem focus of southwest Cameroon

A new index for the risk for transmission of human African trypanosomiasis was developed from an earlier index by adding terms for the proportion of tsetse infected with Trypanosoma brucei gambiense group 1 and the contribution of animals to tsetse diet. The validity of the new index was then assessed in the Fontem focus of southwest Cameroon. Averages of 0.66 and 4.85 Glossina palpalis palpalis (Diptera: Glossinidae) were caught per trap/day at the end of one rainy season (November) and the start of the next (April), respectively. Of 1596 tsetse flies examined, 4.7% were positive for Trypanosoma brucei s.l. midgut infections and 0.6% for T. b. gambiense group 1. Among 184 bloodmeals identified, 55.1% were from pigs, 25.2% from humans, 17.6% from wild animals and 1.2% from goats. Of the meals taken from humans, 81.5% were taken at sites distant from pigsties. At the end of the rainy season, catches were low and similar between biotopes distant from and close to pigsties, but the risk for transmission was greatest at sites distant from the sties, suggesting that the presence of pigs reduced the risk to humans. At the beginning of the rainy season, catches of tsetse and risk for transmission were greatest close to the sties. In all seasons, there was a strong correlation between the old and new indices, suggesting that both can be used to estimate the level of transmission, but as the new index is the more comprehensive, it may be more accurate.     

New tools for the study of animal trypanosomiasis in the Sudan: model-building of dangerous epidemiological passage by remote sensing geographic information systems

Recent studies in a rangeland area of Burkina Faso showed that riparian tsetse flies (Glossina tachinoides and G. palpalis gambiensis) were found along the main rivers, but depending on their location, they had different hosts and were not infected by the same trypanosomoses. There were different epidemiological situations within a distance of a few kilometres, and local assessment of the trypanosome risk thus called for a global approach taking account of the environmental and human factors involved in the interfaces between hosts and vectors. Various types of information concerning entomology, parasitology, ecology, land occupation and animal production systems were fed into a Geographical Information System. High spatial resolution remote sensing tools and original modelling methods were used to detect the valley landscapes most favourable to tsetse flies, and describe land use by herds. The impact of trypanosomes appeared to depend largely on animal movements, watering practices and the degree of contact with riparian tsetse flies. Linking these types of information revealed the most dangerous sites in epidemiological terms, which in this case represented some 18% of the network initially surveyed.