Glossina Austeni Research Articles

Putative bloodmeal sources in Glossina austeni tsetse fly of Arabuko Sokoke National Reserve in Kenya.

Tsetse flies, the sole biological vectors of trypanosomiasis, are predominantly controlled using visual traps and targets baited with attractant lures. Formulation of the lures is informed by compositions of odors from vertebrate hosts preferred by specific tsetse species. However, there are no effective lures for Glossina austeni, a major vector of trypanosomiasis along eastern-coastal region of Africa. Formulation of the lure can be informed by knowledge of G. austeni, preferred vertebrate hosts. We thus sought to understand these hosts by assessment of putative bloodmeal sources of this tsetse fly in Arabuko Sokoke National Reserve where this species is naturally present. We sampled tsetse flies using NGU traps, isolated non-teneral G. austeni flies based on their feeding status, and identified vertebrate source of bloodmeals in their midgut contents using vertebrate 16S rRNA-PCR High-Resolution Melting analysis. We analyzed the relative vertebrate species frequencies in the bloodmeals using Fisher's exact tests. Overall, we trapped 122 flies, most of which (66.39%) were non-teneral, among which we successfully identified the vertebrate bloodmeals in 30 samples. Specifically, we detected putative suni antelope (Neotragus moschatus), harnessed bushbuck (Tragelaphus scriptus), buffalo (Syncerus caffer) and cattle (Bos taurus) derived bloodmeals. Putative suni antelope bloodmeals were significantly more frequent (63.22%), than those of the harnessed bushbuck (23.33%), buffalo (10.00%) or cattle (3.33%) (p < 0.05 Fisher's exact tests) among the samples analyzed. Suni antelope thus appears to predominate vertebrate bloodmeal source for G. austeni in the reserve, coincident with findings reported elsewhere, and is therefore a viable candidate for bioprospecting for G. austeni responsive attractants.

Interactions between Glossina pallidipes salivary gland hypertrophy virus and tsetse endosymbionts in wild tsetse populations

BackgroundTsetse control is considered an effective and sustainable tactic for the control of cyclically transmitted trypanosomosis in the absence of effective vaccines and inexpensive, effective drugs. The sterile insect technique (SIT) is currently used to eliminate tsetse fly populations in an area-wide integrated pest management (AW-IPM) context in Senegal. For SIT, tsetse mass rearing is a major milestone that associated microbes can influence. Tsetse flies can be infected with microorganisms, including the primary and obligate Wigglesworthia glossinidia, the commensal Sodalis glossinidius, and Wolbachia pipientis. In addition, tsetse populations often carry a pathogenic DNA virus, the Glossina pallidipes salivary gland hypertrophy virus (GpSGHV) that hinders tsetse fertility and fecundity. Interactions between symbionts and pathogens might affect the performance of the insect host.MethodsIn the present study, we assessed associations of GpSGHV and tsetse endosymbionts under field conditions to decipher the possible bidirectional interactions in different Glossina species. We determined the co-infection pattern of GpSGHV and Wolbachia in natural tsetse populations. We further analyzed the interaction of both Wolbachia and GpSGHV infections with Sodalis and Wigglesworthia density using qPCR.ResultsThe results indicated that the co-infection of GpSGHV and Wolbachia was most prevalent in Glossina austeni and Glossina morsitans morsitans, with an explicit significant negative correlation between GpSGHV and Wigglesworthia density. GpSGHV infection levels > 103.31 seem to be absent when Wolbachia infection is present at high density (> 107.36), suggesting a potential protective role of Wolbachia against GpSGHV.ConclusionThe result indicates that Wolbachia infection might interact (with an undefined mechanism) antagonistically with SGHV infection protecting tsetse fly against GpSGHV, and the interactions between the tsetse host and its associated microbes are dynamic and likely species specific; significant differences may exist between laboratory and field conditions.Graphical

Gamma Irradiation and Male Glossina austeni Mating Performance.

Simple SummaryAfrican animal trypanosomosis, also known as nagana, is a neglected disease in South Africa, transmitted by tsetse flies. It has been proposed to manage this disease through the eradication of Glossina austeni, one of the vectors of this disease in South Africa. The strategy would be an integration of various control tactics and the release of sterilised colonised males (sterile insect technique) to eliminate relic pockets. The irradiated colonised males must be able to compete with wild males. In preparation of the technique, the mating performance of colony-reared male flies was assessed. Factors that can influence male mating performance, such as radiation dose, and the development stage that is exposed to radiation were evaluated in the laboratory and under semi-field conditions. Radiation doses of 80 Gy and 100 Gy induced 97–99% sterility in colony females that mated with colony males treated as adults or pupae. Walk-in field cage assessments indicated that a dose of up to 100 Gy did not adversely affect the mating performance of males irradiated as adults or late stage pupae. This study indicated that the colonized G. austeni males irradiated as adults or late stage pupae will be suited for the sterile insect technique. An area-wide integrated pest management (AW-IPM) strategy with a sterile insect technique (SIT) component has been proposed for the management of African animal trypanosomosis (AAT) in South Africa. In preparation for the SIT, the mating performance of colony reared Glossina austeni males under influencing factors such as radiation dose and the development stage that is exposed to radiation, was assessed under laboratory and semi-field conditions. The radiation sensitivity of G. austeni colonized 37 years ago when treated as adults and late-stage pupae was determined. Radiation doses of 80 Gy and 100 Gy induced 97–99% sterility in colony females that mated with colony males treated as adults or pupae. Males irradiated either as adults or pupae with a radiation dose of 100 Gy showed similar insemination ability and survival as untreated males. Walk-in field cage assessments indicated that a dose of up to 100 Gy did not adversely affect the mating performance of males irradiated as adults or late stage pupae. Males irradiated as adults formed mating pairs faster than fertile males and males irradiated as pupae. The mating performance studies indicated that the colonized G. austeni males irradiated as adults or late stage pupae will still be suited for SIT.

Expansions of chemosensory gene orthologs among selected tsetse fly species and their expressions in Glossina morsitans morsitans tsetse fly.

Tsetse fly exhibit species-specific olfactory uniqueness potentially underpinned by differences in their chemosensory protein repertoire. We assessed 1) expansions of chemosensory protein orthologs in Glossina morsitans morsitans, Glossina pallidipes, Glossina austeni, Glossina palpalis gambiensis, Glossina fuscipes fuscipes and Glossina brevipalpis tsetse fly species using Café analysis (to identify species-specific expansions) and 2) differential expressions of the orthologs and associated proteins in male G. m. morsitans antennae and head tissues using RNA-Seq approaches (to establish associated functional molecular pathways). We established accelerated and significant (P<0.05, λ = 2.60452e-7) expansions of gene families in G. m. morsitans Odorant receptor (Or)71a, Or46a, Ir75a,d, Ionotropic receptor (Ir) 31a, Ir84a, Ir64a and Odorant binding protein (Obp) 83a-b), G. pallidipes Or67a,c, Or49a, Or92a, Or85b-c,f and Obp73a, G. f. fuscipes Ir21a, Gustatory receptor (Gr) 21a and Gr63a), G. p. gambiensis clumsy, Ir25a and Ir8a, and G. brevipalpis Ir68a and missing orthologs in each tsetse fly species. Most abundantly expressed transcripts in male G. m. morsitans included specific Or (Orco, Or56a, 65a-c, Or47b, Or67b, GMOY012254, GMOY009475, and GMOY006265), Gr (Gr21a, Gr63a, GMOY013297 and GMOY013298), Ir (Ir8a, Ir25a and Ir41a) and Obp (Obp19a, lush, Obp28a, Obp83a-b Obp44a, GMOY012275 and GMOY013254) orthologs. Most enriched biological processes in the head were associated with vision, muscle activity and neuropeptide regulations, amino acid/nucleotide metabolism and circulatory system processes. Antennal enrichments (>90% of chemosensory transcripts) included cilium-associated mechanoreceptors, chemo-sensation, neuronal controlled growth/differentiation and regeneration/responses to stress. The expanded and tsetse fly species specific orthologs includes those associated with known tsetse fly responsive ligands (4-methyl phenol, 4-propyl phenol, acetic acid, butanol and carbon dioxide) and potential tsetse fly species-specific responsive ligands (2-oxopentanoic acid, phenylacetaldehyde, hydroxycinnamic acid, 2-heptanone, caffeine, geosmin, DEET and (cVA) pheromone). Some of the orthologs can potentially modulate several tsetse fly species-specific behavioral (male-male courtship, hunger/host seeking, cool avoidance, hygrosensory and feeding) phenotypes. The putative tsetse fly specific chemosensory gene orthologs and their respective ligands provide candidate gene targets and kairomones for respective downstream functional genomic and field evaluations that can effectively expand toolbox of species-specific tsetse fly attractants, repellents and other tsetse fly behavioral modulators.

Using genetic and phenetic markers to assess population isolation within the southernmost tsetse fly belt in Africa.

The effective control of tsetse flies (Diptera; Glossinidae), the biological vectors of trypanosome parasites that cause human African trypanosomosis and African animal trypanosomosis throughout sub-Saharan Africa, is crucial for the development of productive livestock systems. The degree of genetic isolation of the targeted populations, which indicate reinvasion potential from uncontrolled areas, will be critical to establish a control strategy. Molecular and morphometrics markers were used to assess the degree of genetic isolation between seemingly fragmented populations of Glossina brevipalpis Newstead and Glossina austeni Newstead present in South Africa. These populations were also compared with flies from adjacent areas in Mozambique and Eswatini. For the molecular markers, deoxyribonucleic acid was extracted, a r16S2 Polymerase chain reaction (PCR) was performed and the PCR product sequenced. Nine landmarks were used for the morphometrics study as defined by vein intersections in the right wings of female flies. Generalised Procrustes analyses and regression on centroid size were used to determine the Cartesian coordinates for comparison between populations. Both methods indicated an absence of significant barriers to gene flow between the G. brevipalpis and G. austeni populations of South Africa and southern Mozambique. Sustainable control can only be achieved if implemented following an area-wide management approach against the entire G. brevipalpis and G. austeni populations of South Africa and southern Mozambique. Limited gene flow detected between the G. austeni population from Eswatini and that of South Africa or Mozambique may imply that these two populations are in the proses of becoming isolated.

African animal trypanosomosis (nagana) in northern KwaZulu-Natal, South Africa: Strategic treatment of cattle on a farm in endemic area

African animal trypanosomosis (AAT) is caused by several species of the genus Trypanosoma, a parasitic protozoan infecting domestic and wild animals. One of the major effects of infection with pathogenic trypanosome is anaemia. Currently, the control policies for tsetse and trypanosomosis are less effective in South Africa. The only response was to block treat all infected herds and change the dip chemical to one which controls tsetse flies during severe outbreaks. This policy proved to be less effective as demonstrated by the current high level of trypanosome infections in cattle. Our objective was to study the impacts of AAT (nagana) on animal productivity by monitoring the health of cattle herds kept in tsetse and trypanosomosis endemic areas before and after an intervention that reduces the incidence of the disease. The study was conducted on a farm in northern KwaZulu-Natal which kept a commercial cattle herd. There was no history of any cattle treatment for trypanosome. All cattle were generally in poor health condition at the start of the study though the herd received regular anthelminthic treatment. A treatment strategy using two drugs, homidium bromide (ethidium) and homidium chloride (novidium), was implemented. Cattle were monitored regularly for 13 months for herd trypanosomosis prevalence (HP), herd average packed cell volume (H-PCV) and the percentage of the herd that was anaemic (HA). A total of six odour-baited H-traps were deployed where cattle grazed from January 2006 to August 2007 to monitor the tsetse population. Glossina brevipalpis Newstead and Glossina austeni Newstead were collected continuously for the entire study period. High trypanosomes HP (44%), low average H-PCV (29.5) and HA (24%) were rerecorded in the baseline survey. All cattle in the herd received their first treatment with ethidium bromide. Regular monthly sampling of cattle for the next 142 days showed a decline in HP of 2.2% – 2.8%. However, an HP of 20% was recorded by day 220 and the herd received the second treatment using novidium chloride. The HP dropped to 0.0% and HA to 0.0% by day 116 after the second treatment. The cow group was treated again by day 160 when the HP and HA were 27.3% and 11%, respectively. The same strategy was applied to the other two groups of weaners and the calves at the time when their HP reached 20%. Ethidium and novidium treatment protected cattle, that were under continuous tsetse and trypanosomosis challenge, for up to 6 months. Two to three treatments per year may be sufficient for extended protection. However, this strategy would need to be included into an integrated pest management approach combining vector control for it to be sustainable.

Improving the Diet for the Rearing of Glossina brevipalpis Newstead and Glossina austeni Newstead: Blood Source and Collection - Processing - Feeding Procedures.

One of the challenges to maintain tsetse fly (Diptera: Glossinidae) colonies is the sustainable supply of high quality blood meals. The effect of using anticoagulants during collection of the blood, the addition of phagostimulants to the blood meals as well as using mixtures of bovine and porcine blood in different proportions for feeding on colony productivity was assessed. Defibrinated bovine blood was found to be suitable to maintain both the Glossina brevipalpis Newstead and Glossina austeni Newstead colonies. Blood collected with the anticoagulants sodium citrate, citric sodium combination, citrate phosphate dextrose adenine and citric acid did not affect colony performance of both species. Defibrinated bovine and porcine blood in a 1:1 ratio or the feeding of either bovine or porcine blood on alternating days improved pupae production of G. austeni and can be used to enhance colony growth. Bovine blood is appropriate to maintain G. brevipalpis colonies, however, feeding either bovine or porcine blood on alternating days did improve productivity. Adding the phagostimulants inosine tri-phosphate, cytosine mono-phosphate and guanosine mono-phosphate to the blood at a concentration of 10−4 M improved pupae production of the G. brevipalpis colony. The addition of adenosine tri-phosphate and inosine tri-phosphate improved the performance of the G. austeni colony. Decisions on the most suitable rearing diet and feeding protocols will not only depend on the biological requirements of the species but also on the continuous supply of a suitable blood source that can be collected and processed in a cost-effective way.

An update of the tsetse fly (Diptera: Glossinidae) distribution and African animal trypanosomosis prevalence in north-eastern KwaZulu-Natal, South Africa

An unpredicted outbreak of African animal trypanosomosis or nagana in 1990 in north-eastern KwaZulu-Natal necessitated an emergency control programme, utilising the extensive cattle-dipping system in the area, as well as a reassessment of the tsetse and trypanosomosis problem in the province. Since 1990, sporadic blood sampling of cattle at the dip tanks in the nagana-infested areas were undertaken to identify trypanosome species involved and to determine the infection prevalence in cattle. The distribution and species composition of the tsetse populations in the area were also investigated. From November 2005 to November 2007 selected dip tanks were surveyed for trypanosome infection prevalence. During April 2005 to August 2009 the distribution and abundance of tsetse populations were assessed with odour-baited H traps. The tsetse and trypanosome distribution maps were updated and potential correlations between tsetse apparent densities (ADs) and the prevalence of trypanosomosis were assessed. Glossina brevipalpis Newstead and Glossina austeni Newstead were recorded in locations where they have not previously been collected. No significant correlation between tsetse relative abundance and nagana prevalence was found, which indicated complex interactions between tsetse fly presence and disease prevalence. This was epitomised by data that indicated that despite large differences in the ADs of G. austeni and G. brevipalpis, trypanosome infection prevalence was similar in all three districts in the area. This study clearly indicated that both tsetse species play significant roles in trypanosome transmission and that it will be essential that any control strategy, which aims at sustainable management of the disease, should target both species.

Genome-Wide Comparative Analysis of Chemosensory Gene Families in Five Tsetse Fly Species

For decades, odour-baited traps have been used for control of tsetse flies (Diptera; Glossinidae), vectors of African trypanosomes. However, differential responses to known attractants have been reported in different Glossina species, hindering establishment of a universal vector control tool. Availability of full genome sequences of five Glossina species offers an opportunity to compare their chemosensory repertoire and enhance our understanding of their biology in relation to chemosensation. Here, we identified and annotated the major chemosensory gene families in Glossina. We identified a total of 118, 115, 124, and 123 chemosensory genes in Glossina austeni, G. brevipalpis, G. f. fuscipes, G. pallidipes, respectively, relative to 127 reported in G. m. morsitans. Our results show that tsetse fly genomes have fewer chemosensory genes when compared to other dipterans such as Musca domestica (n>393), Drosophila melanogaster (n = 246) and Anopheles gambiae (n>247). We also found that Glossina chemosensory genes are dispersed across distantly located scaffolds in their respective genomes, in contrast to other insects like D. melanogaster whose genes occur in clusters. Further, Glossina appears to be devoid of sugar receptors and to have expanded CO2 associated receptors, potentially reflecting Glossina's obligate hematophagy and the need to detect hosts that may be out of sight. We also identified, in all species, homologs of Ir84a; a Drosophila-specific ionotropic receptor that promotes male courtship suggesting that this is a conserved trait in tsetse flies. Notably, our selection analysis revealed that a total of four gene loci (Gr21a, GluRIIA, Gr28b, and Obp83a) were under positive selection, which confers fitness advantage to species. These findings provide a platform for studies to further define the language of communication of tsetse with their environment, and influence development of novel approaches for control.

Evaluation of larviposition site selection of Glossina brevipalpis

Tsetse species (Diptera: Glossinidae) are vectors of trypanosome parasites which cause disease in both humans and livestock. In South Africa Glossina austeni Newstead, 1912 and G. brevipalpis Newstead, 1911 are responsible for the cyclical transmission of animal trypanosomes causing African animal trypanosomiasis also referred to as nagana. Gravid tsetse females deposit a single larva in specific sites but little information is available on biotic and abiotic factors that govern site selection. This study therefore aimed to characterize some of the substrate conditions that may influence selection of larviposition sites. Colonised, gravid female G. brevipalpis were presented with a choice of four larviposition sites. Sites differed in qualities of pH (5, 7, 9), salinity (0, 1.3, 4g/L) and the presence of other tsetse pupae (G. brevipalpis or G. austeni). These trials indicated no significant selection by gravid females with regard to pH and salinity. Females selected significantly more often for sites with pupae (P<0.05), but also favored sites containing conspecific over heterospecific pupae (P<0.05). These results present the first indication of an aggregation effect of tsetse pupae in G. brevipalpis. This may imply that G. brevipalpis larvae produce a pheromone during pupation as seen in G. morsitans morsitans. Isolation of such semio-chemicals would allow the development of larviposition traps to attract gravid females.

Determination of the optimal mating age of colonised Glossina brevipalpis and Glossina austeni using walk-in field cages in South Africa.

BackgroundFor the control of Glossina brevipalpis and Glossina austeni that occur in South Africa an area-wide integrated pest management (AW-IPM) program with a sterile insect technique (SIT) component has been proposed. The quality of the released sterile male tsetse flies will greatly determine the success of the SIT component of the programme. Sterile males need to be able to compete with wild males immediately after their release in the affected area. The mating competitiveness can be affected by many factors including the optimal mating age of the fly which can have an impact on the timing of the release.MethodsTo assess the optimal mating age for G. brevipalpis and G. austeni, mating competitiveness studies were carried out in a walk-in field cage. First, the time of peak fly activity was determined by performing the experiment in the morning and then again in the afternoon. Thereafter, 3, 6 and 9-day-old male flies competed for 3-day-old virgin females.ResultsThere were no significant differences in mating performance when the field cage experiments were done in the morning or in the afternoon. However, the mating latency was shorter in the afternoon than in the morning. For both species 9-day-old males mated significantly more often than 6 or 3-day-old males. Age did not affect the males’ ability to transfer sperm, mating duration or the mating latency. All females that mated were inseminated.ConclusionsAge did influence the mating competitiveness of G. brevipalpis and G. austeni and it is recommended that sterile males are not released before the age of 9 days. Keeping the male flies in the rearing facility for 8 days will have economic and logistic consequences for AW-IPM programmes that have a SIT component.

Bovine trypanosomosis prevalence at the edge of Hluhluwe-iMfolozi Park, KwaZulu-Natal, South Africa.

The northern KwaZulu-Natal (NKZN) region of South Africa is the southern limit of the African tsetse belt. Entomological information on Glossina brevipalpis and Glossina austeni was generated following the outbreak of trypanosomosis in cattle in 1990. However, these data have not been supported by parallel studies on epidemiology of the disease and therefore there has been no control policy in place. This study presented the first intensive investigations to address the epidemiology of trypanosomosis in NKZN. Tsetse abundance, trypanosome herd average prevalence (HAP), herd average anaemia (HAA) and herd average packed cell volume (HA-PCV) were investigated at three communal diptanks located at the edge of Hluhluwe-iMfolozi Park by monthly sampling from June 2006 - November 2007. Seasonal trypanosome surveys were conducted at seven other communal diptanks. Glossina brevipalpis prevalence was high at two of the diptanks, Mvutshini and Ekuphindisweni, but low at Ocilwane, whilst G. austeni was only collected from Mvutshini. This high and low tsetse challenge presented different disease scenarios. Cattle at Mvutshini and Ekuphindisweni had the highest HAP of 12.3% and 8.9% respectively, both significantly different (p = 0.001) from the HAP obtained from cattle at Ocilwane (2.9%). These two cattle herds also had the highest HAA, 27.7% and 33.4% respectively, whilst cattle at Ocilwane had the lowest, 11.1% (p = 0.001). Conversely, cattle at Ocilwane had the highest HA-PCV, ranging between 29.0% and 32.0%, whilst cattle at Mvutshini and Ekuphindisweni had the lowest HA-PCV (24.0% - 29.0%). By combining the data from the three diptanks (1318 observations), 62.0% of the infected cattle were found anaemic, compared to 20.0% in the uninfected group. Trypanosome seasonal surveys showed that cattle at all the seven diptanks were infected with trypanosomes; mean HAP, HAA and HA-PCV of 10.2%, 46.6% and 23.7%, respectively. This study generated information on the epidemiological factors related to the wide spread of trypanosome-infected cattle and tsetse flies. Trypanosomosis is a disease of economic importance impacting the livelihood of resource-poor farmers in NKZN.

Vector competence of &lt;i&gt;Glossina austeni&lt;/i&gt; and &lt;i&gt;Glossina brevipalpis&lt;/i&gt; for &lt;i&gt;Trypanosoma congolense&lt;/i&gt; in KwaZulu-Natal, South Africa

Tsetse-transmitted trypanosomosis (nagana) has been the cause of stock losses in the recent past and still presents a major problem to livestock owners in certain areas of KwaZulu- Natal, South Africa. Over 10 000 cattle mortalities were reported in the 1990 nagana outbreak. Although information on the distribution and abundance of the tsetse flies Glossina brevipalpis and Glossina austeni in KwaZulu-Natal exists, data on their vector competence are lacking. This study aimed to determine the rate of natural Trypanosoma congolense infection by field-collected as well as colony-reared flies of these species. A total of 442 field-collected G. brevipalpis and 40 G. austeni flies were dissected immediately after collection to determine their infection rates, whilst 699 G. brevipalpis and 49 G. austeni flies were fed on susceptible animals in 10 and four batches, respectively, for use in xenodiagnosis experiments. Teneral colony flies were fed on infected animals and dissected 21 days post infection to confirm their infectivity testing. Glossina austeni harboured 8% immature and mature infections. In G. brevipalpis, the infection with the immature stages was lower (1%) and no mature infections were observed. Although all four batches of G. austeni transmitted T. congolense to four susceptible animals, no transmission resulted from 10 batches of G. brevipalpis fed on susceptible cattle. Colony-derived G. austeni (534) and G. brevipalpis (882) were fed on four bovines infected with different T. congolense isolates. Both G. austeni and G. brevipalpis acquired trypanosome infection from the bovines, with immature infection ranges of 20% - 33% and 1% - 4%, respectively. Parasites, however, only matured in G. austeni (average = 4%). Glossina austeni plays a larger role in the epidemiology of animal trypanosomosis in KwaZulu-Natal than G. brevipalpis and therefore more focus should be aimed at the former when control measures are implemented.

Theoretical levels of control as a function of mean temperature and spray efficacy in the aerial spraying of tsetse fly

The hypothetical impact of aerial spraying on tsetse fly populations is investigated. Spray cycles are scheduled at intervals two days short of the first interlarval period and halted once the last of the female flies that originated from pre-spray-deposited pupae have been sprayed twice. The effect of temperature on the aerial spraying of tsetse, through its reproductive cycle and general population dynamics, is of particular interest, given that cooler weather is preferred for the settling of insecticidal droplets. Spray efficacy is found to come at a price due to the greater number of cycles necessitated by cooler weather. The extra cost is argued to be worth while. Pupae, still in the ground at the end of spraying, are identified as the main threat to a successful operation. They are slightly more vulnerable at the low temperature extreme of tsetse habitat (16°C), when the cumulative, natural pupal mortality is high. One can otherwise base one's expectations on the closeness with which the time to the third last spray approaches one puparial duration. A disparity of anything close to the length of a spray cycle advocates caution, whereas one which comes close to vanishing should be interpreted as being auspicious. Three such key temperatures, just below which one can anticipate an improved outcome and just above which caution should be exercised, are 17.146°C, 19.278°C and 23.645°C. A refinement of the existing formulae for the puparial duration and the first interlarval period might be prudent in the South African context of a sympatric Glossina brevipalpis–Glossina austeni, tsetse population. The resulting aerial spraying strategy would then be formulated using a G. brevipalpis puparial duration and a G. austeni first interlarval period.

The finite element implementation of a K.P.P. equation for the simulation of tsetse control measures in the vicinity of a game reserve

An equation, strongly reminiscent of Fisher’s equation, is used to model the response of tsetse populations to proposed control measures in the vicinity of a game reserve. The model assumes movement is by diffusion and that growth is logistic. This logistic growth is dependent on an historical population, in contrast to Fisher’s equation which bases it on the present population. The model therefore takes into account the fact that new additions to the adult fly population are, in actual fact, the descendents of a population which existed one puparial duration ago, furthermore, that this puparial duration is temperature dependent. Artificially imposed mortality is modelled as a proportion at a constant rate. Fisher’s equation is also solved as a formality. The temporary imposition of a 2 % day −1 mortality everywhere outside the reserve for a period of 2 years will have no lasting effect on the influence of the reserve on either the Glossina austeni or the G. brevipalpis populations, although it certainly will eradicate tsetse from poor habitat, outside the reserve. A 5 km-wide barrier with a minimum mortality of 4 % day −1, throughout, will succeed in isolating a worst-case, G. austeni population and its associated trypanosomiasis from the surrounding areas. A more optimistic estimate of its mobility suggests a mortality of 2 % day −1 will suffice. For a given target-related mortality, more mobile species are found to be more vulnerable to eradication than more sedentary species, while the opposite is true for containment.

The Glossina proteolytic lectin (Gpl) gene is expressed only in members of Glossina species

Differentiation of bloodstream-form trypanosomes into procyclics in tsetse flies (Diptera: Glossinidae) is a crucial step in the establishment of midgut infections. A number of factors have been implicated in the transformation process, including enzymes and lectins or lectin-like molecules. Recently, Glossina proteolytic lectin (Gpl) gene, which encodes a protein with both lectin and trypsin activities has been shown to stimulate transformation of bloodstream-form trypanosomes into procyclics in vitro. Using RT-PCR, we show that the induction of Gpl gene expression by blood meal occurs only in Glossina fuscipes fuscipes Newstead, Glossina austeni Newstead, Glossina pallidipes Austen, and not in the Anopheles gambiae Giles sensu stricto, Phlebotomus duboscqi Neveu-Lemaire, Rhipicephalus appendiculatus Neumann and Stomoxys calcitrans (Linnaeus). The expression means of Gpl mRNA in G. f. fuscipes following a blood meal were significant (P , 0.05) with low expression in teneral flies and reaching a maximum between 48 and 72 h (P , 0.05), suggesting time-dependent regulation of the transcription. The expression of the Gpl gene was significantly lower (P , 0.05) in G. f. fuscipes fed on blood meal infected with Trypanosoma brucei brucei as compared with G. f. fuscipes fed on uninfected blood meal. This suggests some form of interaction of T. b. brucei or the parasite products with Gpl within the tsetse midgut leading to down-regulation of the Gpl gene. Additionally, refractory G. f. fuscipes expressed higher (P , 0.05) transcript abundance than the susceptible G. pallidipes.

Evaluation and improvement of sticky traps as monitoring tools for Glossina austeni and G. brevipalpis (Diptera: Glossinidae) in north-eastern KwaZulu-Natal, South Africa

The attractiveness of various colours, colour combinations and sizes of sticky traps of the 3-dimensional trap (3DT), cross-shaped target (XT), rectangular screen (RT) and monopanels were evaluated for their efficacy to capture Glossina austeni Newstead and G. brevipalpis Newstead in north-eastern KwaZulu-Natal, South Africa. The 3-dimensional shapes of the XT and 3DT in light blue (l.blue) and white were significantly (ca. 3.1-6.9 times) better than the RT for G. austeni. On bicoloured XTs, G. austeni landed preferentially on electric blue (e.blue) (58%) and black (63%) surfaces when used with white; while for G. brevipalpis, significantly more landed on e.blue (60-66%) surfaces when used with l.blue, black or white surfaces. Increased trap size increased the catches of G. brevipalpis females and both sexes of G. austeni significantly. Temoocid and polybutene sticky materials were equally effective and remained durable for 2-3 weeks. The glossy shine of trap surfaces did not have any significant effect on the attraction and landing responses of the two species. The overall trap efficiency of the e.blue/l.blue XT was 23% for G. brevipalpis and 28% for G. austeni, and that of the e.blue/black XT was 16% for G. brevipalpis and 51% for G. austeni. Larger monopanels, painted e.blue/black on both sides, increased the catches of G. austeni females significantly by up to four times compared to the standard e.blue/black XT. This monopanel would be recommended for use as a simple and cost effective survey tool for both species in South Africa.

Livestock farmers’ perception and epidemiology of bovine trypanosomosis in Kwale District, Kenya

We did cross-sectional surveys in Kwale District, Kenya to determine the epidemiology of bovine trypanosomosis and livestock owners’ perceptions of the disease. The surveys involved relative importance of trypanosomosis, examination of the current disease constraints, current control practices and drug-use patterns. Informal meetings were held with farmers and cattle census undertaken. Tsetse-fly densities and trypanosomosis prevalences in cattle were determined. A total of 132 farmers were interviewed. Trypanosomosis, anaplasmosis, East Coast fever, foot-and-mouth diseases were reported to be the major constraints to livestock production. Trypanosomosis was the most important compared to other diseases. Chemotherapy was the most widely used method of controlling the disease. Farmer-based tsetse-control technologies were poorly adopted. Respondents were quite knowledgeable on the symptoms, causes and treatment of trypanosomosis. Glossina austeni, G. brevipalpis and G. pallidipes were found in the area; the latter was the most common (0.2–738 flies/trap). Trypanosoma congolense and T. vivax were found in cattle with the former more prevalent. Infection prevalences in cattle varied between 0 and 25% (median: 22%).

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

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

Polytene chromosome maps in four species of tsetse flies Glossina austeni, G. pallidipes, G. morsitans morsitans and G. m. submorsitans (Diptera: Glossinidae): a comparative analysis

Photographic polytene chromosome maps from pupal trichogen cells of four tsetse species, Glossina austeni, G. pallidipes, G. morsitans morsitans and G. m. submorsitans were constructed and compared. The homology of chromosomal elements between the species was achieved by comparing banding patterns. The telomeric and subtelomeric chromosome regions were found to be identical in all species. The pericentromeric regions were found to be similar in the X chromosome and the left arm of L1 chromosome (L1L) but different in L2 chromosome and the right arm of L1 chromosome (L1R). The L2 chromosome differs by a pericentric inversion that is fixed in the three species, G. pallidipes, G. morsitans morsitans and G. m. submorsitans. Moreover, the two morsitans subspecies appeared to be homosequential and differ only by two paracentric inversions on XL and L2L arm. Although a degree of similarity was observed across the homologous chromosomes in the four species, the relative position of specific chromosome regions was different due to chromosome inversions established during their phylogeny. However, there are regions that show no apparent homology between the species, an observation that may be attributed to the considerable intra--chromosomal rearrangements that have occurred following the species divergence. The results of this comparative analysis support the current phylogenetic relationships of the genus Glossina.