Tsetse Fly Species Research Articles

Tsetse fly density and trypanosoma infection rate in Bedele and Dabo Hana districts of Buno Bedele Zone, Southwest Ethiopia

BackgroundTrypanosomiasis is an infectious disease caused by parasitic protozoa of the genus Trypanosome and primarily transmitted by tsetse flies. This study aimed to determine the density of tsetse flies and the rate of trypanosome infection in the Bedele and Dabo Hana districts of the Buno Bedele Zone in Ethiopia.ResultsA cross-sectional study was conducted from January to February 2023 to catch tsetse flies, determine tsetse density, and estimate the trypanosome infection rate. We used 100 traps (40 NGU, 30 pyramidal, and 30 biconical) to catch the flies. The following standard procedures were followed to identify the specific trypanosome species in the collected tsetse flies: The flies were dissected, and the salivary glands were removed. We placed the salivary glands in a drop of saline solution on a microscope slide. A coverslip was placed over the salivary glands, the slide was examined under a microscope, and the trypanosomes were identified based on their morphology. A total of 3,740 tsetse flies were captured from 100 traps, resulting in an overall apparent density of 18.7 flies per trap per day. Within the study area, only one species of tsetse fly, Glossina tachinoides, was identified. Of the 1,320 dissected Glossina tachinoides, 1.82% were found to be infected with trypanosome parasites. Among these infections, 58.33% were attributed to Trypanosoma congolense, while the remaining 41.67% were caused by Trypanosoma brucei. The infection rate of trypanosomes was significantly higher in female tsetse flies (87.5%) as compared to male flies (12.5%). Furthermore, a significantly higher infection rate was observed in flies older than 20 days (83.33%) and in hunger stage 1 flies (58.33%) compared to hunger stages 2, 3, and 4.ConclusionsThis study highlights the necessity of implementing control and suppression measures targeting the vector (tsetse flies) and the parasite (trypanosomes) to effectively manage and prevent pathogenic animal trypanosomiasis.

An Overview of Tsetse Fly Repellents: Identification and Applications.

Tsetse flies are vectors of the parasite trypanosoma that cause the neglected tropical diseases human and animal African trypanosomosis. Semiochemicals play important roles in the biology and ecology of tsetse flies. Previous reviews have focused on olfactory-based attractants of tsetse flies. Here, we present an overview of the identification of repellents and their development into control tools for tsetse flies. Both natural and synthetic repellents have been successfully tested in laboratory and field assays against specific tsetse fly species. Thus, these repellents presented as innovative mobile tools offer opportunities for their use in integrated disease management strategies.

Modelled impact of Tiny Targets on the distribution and abundance of riverine tsetse.

The insecticide-treated baits known as Tiny Targets are one of the cheapest means of controlling riverine species of tsetse flies, the vectors of the trypanosomes that cause sleeping sickness in humans. Models of the efficacy of these targets deployed near rivers are potentially useful in planning control campaigns and highlighting the principles involved. To evaluate the potential of models, we produced a simple non-seasonal model of the births, deaths, mobility and aging of tsetse, and we programmed it to simulate the impact of seven years of target use against the tsetse, Glossina fuscipes fuscipes, in the riverine habitats of NW Uganda. Particular attention was given to demonstrating that the model could explain three matters of interest: (i) good control can be achieved despite the degradation of targets, (ii) local elimination of tsetse is impossible if invasion sources are not tackled, and (iii) with invasion and target degradation it is difficult to detect any effect of control on the age structure of the tsetse population. Despite its simplifications, the model can assist planning and teaching, but allowance should be made for any complications due to seasonality and management challenges associated with greater scale.

The internal transcribed spacer 1 sequence polymorphism brings updates to tsetse species distribution in the northern Cameroon: Importance in planning efficient vector control.

Vector control remains one of the best strategies to prevent the transmission of trypanosome infections in humans and livestock and, thus, a good way to achieve the elimination of human African trypanosomiasis and animal African trypanosomiasis. A key prerequisite for the success of any vector control strategy is the accurate identification and correct mapping of tsetse species. In this work, we updated the tsetse fly species identification and distribution in many geographical areas in Cameroon. Tsetse flies were captured from six localities in Cameroon, and their species were morphologically identified. Thereafter, DNA was extracted from legs of each tsetse fly and the length polymorphism of internal transcribed spacer-1 (ITS1) region of each fly was investigated using PCR. ITS1 DNA fragments of each tsetse species were sequenced. The sequences obtained were analysed and compared to those available in GenBank. This enabled to confirm/infirm results of the morphologic identification and then, to establish the phylogenetic relationships between tsetse species. Morphologic features allowed to clearly distinguish all the tsetse species captured in the South Region of Cameroon, that is, Glossina palpalis palpalis, G. pallicera, G. caliginea and G. nigrofusca. In the northern area, G. morsitans submorsitans could also be distinguished from G. palpalis palpalis, G. tachinoides and G. fuscipes, but these three later could not be distinguished with routine morphological characters. The ITS1 length polymorphism was high among most of the studied species and allowed to identify the following similar species with a single PCR, that is, G. palpalis palpalis with 241 or 242 bp and G. tachinoides with 221 or 222 bp, G. fuscipes with 236 or 237 bp. We also updated the old distribution of tsetse species in the areas assessed, highlighting the presence of G. palpalis palpalis instead of G. fuscipes in Mbakaou, or in sympatry with G. morsitans submorsitans in Dodeo (northern Cameroon). This study confirms the presence of G. palpalis palpalis in the Adamawa Region of Cameroon. It highlights the limits of using morphological criteria to differentiate some tsetse species. Molecular tools based on the polymorphism of ITS1 of tsetse flies can differentiate tsetse species through a simple PCR before downstream analyses or vector control planning.

Application of biomolecular techniques on tsetse fly puparia for species identification at larvipostion sites.

Puparia are commonly found in tsetse fly larviposition sites during studies on larval ecology. This chitinous shell is representative of past or ongoing exploitation of these sites by tsetse flies. The morphological characteristics of the puparium are not sufficiently distinctive to allow identification of the species. This study explores the applicability of biomolecular techniques on empty puparia for tsetse fly species identification. Five techniques were compared for DNA extraction from tsetse fly puparia, 1/Chelex® 100 Resin, 2/CTAB, 3/Livak's protocol, 4/DEB + proteinase K and 5/QIAamp® DNA Mini kit, using two homogenisation methods (manual and automated). Using a combination of two primer pairs, Chelex, CTAB, and DEB + K proved the most efficient on fresh puparia with 90, 85, and 70% samples identified, respectively. Shifting from fresh to one- to nine-month-old puparia, the Chelex method gave the best result allowing species identification on puparia up to seven months old. The subsequent testing of the Chelex extraction protocol identified 152 (60%) of 252 field-collected puparia samples at species level. The results show that reliable genetic identification of tsetse flies species can be performed from empty puparia, what can prove of great interest for future ecological studies on larviposition sites. The Chelex technique was the most efficient for DNA extraction, though the age-limit of the samples stood at seven months, beyond which DNA degradation probably compromises the genetic analysis.

How plastic are upper thermal limits? A comparative study in tsetse (family: Glossinidae) and wider Diptera

Critical thermal maximum (CTmax) describes the upper thermal tolerance of an animal where biological functions start to fail. A period of acclimation can enhance CTmax through plasticity, potentially buffering animals from extreme temperatures caused by climate change. Basal and acclimated CTmax vary within and between species and may be explained by traits related to thermal physiology, such as body size and sex. Differences in CTmax have not been established among species of tsetse fly (Glossina spp.), vectors of animal and human African trypanosomiasis. Here, we investigated basal CTmax and its plasticity for five tsetse species following adult acclimation at constant 25 or 30 °C for five days. We then set our findings in context using a meta-analysis on 33 species of Diptera. We find that, of the five tsetse species considered, only Glossina palpalis gambiensis and Glossina brevipalpis exhibited plasticity of CTmax, with an increase of 0.12 °C and 0.10 °C per 1 °C acclimation respectively. Within some species, higher basal CTmax values were associated with larger body size and being female, while variation in plasticity (i.e., response to the acclimation temperature) could not be explained by sex or size. Our broader meta-analysis across Diptera revealed overall CTmax plasticity of 0.06 °C per 1 °C acclimation, versus a similar 0.05 °C mean increase in tsetse. In contrast, there was greater CTmax plasticity in males compared to females in Diptera. Our study highlights that CTmax and its plasticity varies even among closely related species. Broader patterns across groups are not always reflected at a finer resolution; we thus emphasise the need for detailed experimental studies across a wide range of insect species to capture their capacity to cope with rapidly warming temperatures.

No evidence for direct thermal carryover effects on starvation tolerance in the obligate blood-feeder, Glossina morsitans morsitans.

Thermal stress during development can prime animals to cope better with similar conditions in later life. Alternatively, negative effects of thermal stress can persist across life stages and result in poorer quality adults (negative carryover effects). As mean temperatures increase due to climate change, evidence for such effects across diverse taxa is required. Using Glossina morsitans morsitans, a species of tsetse fly and vector of trypanosomiasis, we asked whether (i) adaptive developmental plasticity allows flies to survive for longer under food deprivation when pupal and adult temperatures are matched; or (ii) temperature stress during development persists into adulthood, resulting in a greater risk of death. We did not find any advantage of matched pupal and adult temperature in terms of improved starvation tolerance, and no direct negative carryover effects were observed. There was some evidence for indirect carryover effects-high pupal temperature produced flies of lower body mass, which, in turn, resulted in greater starvation risk. However, adult temperature had the largest impact on starvation tolerance by far: flies died 60% faster at 31°C than those experiencing 25°C, consequently reducing survival time from a median of 8 (interquartile range (IQR) 7-9) to 5 (IQR 5-5.25) days. This highlights differences in temperature sensitivity between life stages, as there was no direct effect of pupal temperature on starvation tolerance. Therefore, for some regions of sub-Saharan Africa, climate change may result in a higher mortality rate in emerging tsetse while they search for their first blood meal. This study reinforces existing evidence that responses to temperature are life stage specific and that plasticity may have limited capacity to buffer the effects of climate change.

Molecular detection of Sodalis glossinidius, Spiroplasma species and Wolbachia endosymbionts in wild population of tsetse flies collected in Cameroon, Chad and Nigeria

BackgroundTsetse flies are cyclical vectors of African trypanosomiasis (AT). The flies have established symbiotic associations with different bacteria that influence certain aspects of their physiology. Vector competence of tsetse flies for different trypanosome species is highly variable and is suggested to be affected by bacterial endosymbionts amongst other factors. Symbiotic interactions may provide an avenue for AT control. The current study provided prevalence of three tsetse symbionts in Glossina species from Cameroon, Chad and Nigeria.ResultsTsetse flies were collected and dissected from five different locations. DNA was extracted and polymerase chain reaction used to detect presence of Sodalis glossinidius, Spiroplasma species and Wolbachia endosymbionts, using species specific primers. A total of 848 tsetse samples were analysed: Glossina morsitans submorsitans (47.52%), Glossina palpalis palpalis (37.26%), Glossina fuscipes fuscipes (9.08%) and Glossina tachinoides (6.13%). Only 95 (11.20%) were infected with at least one of the three symbionts. Among infected flies, six (6.31%) had Wolbachia and Spiroplasma mixed infection. The overall symbiont prevalence was 0.88, 3.66 and 11.00% respectively, for Sodalis glossinidius, Spiroplasma species and Wolbachia endosymbionts. Prevalence varied between countries and tsetse fly species. Neither Spiroplasma species nor S. glossinidius were detected in samples from Cameroon and Nigeria respectively.ConclusionThe present study revealed, for the first time, presence of Spiroplasma species infections in tsetse fly populations in Chad and Nigeria. These findings provide useful information on repertoire of bacterial flora of tsetse flies and incite more investigations to understand their implication in the vector competence of tsetse flies.

Bloodmeal host identities among sympatric Glossina austeni and Glossina pallidipes tsetse flies in Shimba Hills National Reserve, Kwale, Kenya.

Odor from preferred/non-preferred tsetse fly vertebrate hosts have been exploited in R&D of attractants/repellents of the fly for human and livestock protection. Odors from vertebrate hosts of Glossina austeni and Glossina pallidipes tsetse flies can facilitate formulation of novel attractants effective against G. austeni or improvement of existing attractant blends for G. pallidipes. We compared vertebrate blood meal sources of both fly species at Shimba Hills National Reserve, Kenya, to establish putative preferred host of either species, hence potential source of G. austeni or G. pallidipes specific odors. We trapped sympatric adult flies in 2021 and 2022 using NGU traps/sticky panels baited with POCA, collected their blood meals and characterize the meals using HRM vertebrate 16S rRNA- PCR (for host identification), and compared host profiles using GLM and Fisher's exact tests. We collected 168 and 62 sympatric G. pallidipes and G. austeni with bloodmeal, respectively in 2021 and, 230 and 142 respectively in 2022. In 2021, we identified putative hosts of 65.48 and 69.35 % of the G. pallidipes and G. austeni respectively and 82.61 and 80.28%, respectively in 2022. In 2021, we detected harnessed bushbuck, buffalo, common warthog and cattle putative host bloodmeals, and additionally bushpig and suni antelope bloodmeals in 2022. Putative vertebrate bloodmeal sources were significantly different by tsetse fly species (χ2(1, N=457) = 43.215, p < 0.001) and sampling year (χ2(1, N=457) = 8.044, p = 0.005). Frequency of common warthog bloodmeals was higher in G. pallidipes (65.79 %) than G. austeni (38.60%), and that of suni antelope and harnessed bushbuck putative bloodmeals higher in G. austeni (21.05-28.07%) than in G. pallidipes (6.84 - 17.37%) in 2022. There was an apparent change in putative feeding preference/host choices in both fly species between 2021 and 2022. Host bloodmeals in G. pallidipes or G. austeni predominantly from putative harnessed bushbuck, suni antelope or common warthog reveal these vertebrates with potential odors that can be harnessed and formulated into appropriate attractants for respective species and integrated into routine control regiment for G. pallidipes and/or G. austeni.

Tsetse fly ecology and risk of transmission of African trypanosomes related to a protected forest area at a military base in the city of Abidjan, Côte d'Ivoire.

African trypanosomoses, whose pathogens are transmitted by tsetse flies, are a threat to animal and human health. Tsetse flies observed at the military base of the French Forces in Côte d'Ivoire (FFCI base) were probably involved in the infection and death of military working dogs. Entomological and parasitological surveys were carried out during the rainy and dry seasons using "Vavoua" traps to identify tsetse fly species, their distribution, favorable biotopes and food sources, as well as the trypanosomes they harbor. A total of 1185 Glossina palpalis palpalis tsetse flies were caught, corresponding to a high average apparent density of 2.26 tsetse/trap/day. The results showed a heterogeneous distribution of tsetse at the FFCI base, linked to more or less favorable biotopes. No significant variation in tsetse densities was observed according to the season. The overall trypanosomes infection rate according to microscopic observation was 13.5%. Polymerase chain reaction (PCR) analyses confirmed the presence of Trypanosoma vivax and T. congolense forest type, responsible for African animal trypanosomosis. Our findings suggest that there is a risk of introduction and transmission of T. brucei gambiense, responsible for human African trypanosomiasis, on the study site. This risk of transmission of African trypanosomes concerns not only the FFCI base, but also inhabited peripheral areas. Our study confirmed the need for vector control adapted to the eco-epidemiological context of the FFCI base.

Diversity of tsetse flies and trypanosome species circulating in the highly infested cattle rearing area of the Faro and Deo subdivision, Adamawa region, Cameroon

Animal African Trypanosomiasis (AAT) remains an animal health problem in sub-Saharan Africa and in Cameroon in particular. Despite more than 40 years of fighting against AAT in some tsetse infested areas, the disease prevalence is still a concern. Improving the control strategies in different settings requires to understand the current epidemiological situation of AAT. The aim of the present study was to update our knowledge on the diversity of tsetse fauna and trypanosome species in the tsetse infested area of Faro and Deo division, Adamawa region, Cameroon. Tsetse flies were caught using Vavoua trap in two villages and the apparent density per trap (ADP) were estimated. After morphological identification of tsetse fly species, flies were dissected and their midguts recovered. The presence of blood meal residues was recorded. Trypanosomes species were checked in the flies' midguts by microscopy followed by PCR method. The vertebrate taxa on which tsetse flies have taken blood meal were determined using the heteroduplex-PCR method. A total of 338 tsetse flies including 11 teneral flies (10 Glossina palpalis palpalis and 01 G. morsitans submorsitans) and 327 non-teneral were trapped in Mayo Lainde and Tchabal Mbabo. Amongst the caught tsetse flies, of the 327 non-teneral flies, 315 (96.3%) were G. p. palpalis, 8 (2.4%) were G. morsitans submorsitans and 4 (1.2%) G. fuscipes fuscipes. Trypanosome infections including Trypanosoma congolense forest (19.88%) and savanah (2.53%) “types”, T. brucei s.l. (7.30%) and T. vivax (2.85%) were identified in 45.08% of non-teneral flies (32.38% for single infection and 12.70% for mixed infection). Amongst the 54 blood meals identified in tsetse midguts, 41% were from humans, 33% from cattle and 26% from other vertebrate hosts. About 51.9% of blood meals were found with various trypanosome species including 42.6% with T. congolense and 24% with T. brucei s.l. This study revealed the presence of three tsetse taxa and the circulation of four trypanosome taxa in villages of the Faro and Deo division. About 45% of captured tsetse fly are infected with trypanosome species causing AAT. Tsetse flies feed on humans, cattle and many other vertebrates. Strategies to eliminate the vectors must be improved to reduce the pathological impacts of trypanosome infections in this area.

Prevalence of Cattle Trypanosomosis and Temporal Vector Distribution in Jima Arjo District, Upper Didessa Valley, Western Ethiopia.

Trypanosomosis is a protozoan disease, mostly transmitted by the tsetse fly, Glossina species, which causes severe disease of livestock in Ethiopia. The disease is also widespread across the globe especially in sub-Saharan African states. A cross-sectional study with the objectives of determining the prevalence of bovine trypanosomosis and assess the apparent densities of the disease vectors was conducted from October 2019 to July 2020 G.C in Jima Arjo district, east Wollega zone, Oromia regional state, Ethiopia. A parasitological study using conventional buffy coat technique was employed for the determination of prevalence of trypanosomosis and species was identified by Giemsa stain technique, while baited traps were used for the vector survey. The results of parasitological study revealed that the overall prevalence was 36 (8.2%) at 95% CI. From the total trypanosome positive animals 22 (5.0%), 8 (1.82%) and 6 (1.36%) of them harbor T. congolense, T. vivax, and T. brucei, respectively. Relatively higher prevalence (10.91%) was seen in animals with poor body condition than those with medium (7.38%) and good (5.55%), body condition though it is not statistically significant (P > 0.05). Higher infection rate was observed in male 26 (12.26%) than female 10 (4.39%) due to male cattle more exposed to the tsetse fly area or early released from home for drought reason. Out of the total positive animals, only 12 of them were anemic on buffy coat test of sampled blood. This justifies that animal could be positive for trypanosomosis without showing clinical sign of anemia which is the dominant sign in this disease. A total of 2185 vectors trypanosomes were collected among which 1,569 were tsetse flies and 616 were other biting flies. The density of Glossina species was 15.1 fly/trap/day. Glossina morsitans submorsitans and G. tachnoides were the two dominant species of tsetse flies recorded from the area. The present study indicated that tsetse and non-tsetse fly-borne trypanosomosis is a leading bottle neck for production and health of animals in Jima Arjo districts and similar case was found throughout the country which necessitates a coordinated vector and parasite control in order to alleviate the problem of the disease.

Entomological assessment of tsetse-borne trypanosome risk in the Shimba Hills human-wildlife-livestock interface, Kenya.

Shimba Hills is a wildlife area in Kenya and a major focus of tsetse-borne trypanosomes in East Africa. In Shimba Hills, tsetse-borne trypanosomes constrain animal health and smallholder livelihoods. However, epidemiological data to guide hotspot-targeted control of infections are limited. This study assessed the dynamics of tsetse-borne trypanosome risk in Shimba Hills with the objective to describe infection hotspots for targeted control. Tsetse flies (n = 696) collected in field surveys between November 2018 and September 2019 in Shimba Hills were characterized for chronological age and phenotypic sizes and screened for trypanosome and cattle DNA. Entomological inoculation rates for trypanosome risk assessment were derived from the product of fly abundance and molecular rates of vector infection and confirmed cattle bloodmeals in tsetse flies. In addition, cattle health indicators including anemia scores were assessed in contemporaneous parasitological surveys that screened livestock blood samples (n = 1,417) for trypanosome using the buffy-coat technique. Compared with Glossina brevipalpis and G. austeni, G. pallidipes was the most abundant tsetse fly species in Shimba Hills and had a wider spatial distribution and greater likelihood for infectious bites on cattle. The risk of cattle infection was similar along the Shimba Hills human-wildlife-livestock interface and high within one thousand meters of the wildlife reserve boundary. Trypanosomes in tsetse flies were highly diverse and included parasites of wild-suids probably acquired from warthogs in Shimba Hills. Age and phenotypic sizes were similar between tsetse fly populations and did not affect the probability of infection or cattle bloodmeals in the vectors. Anemia was more likely in trypanosome-positive cattle whilst parasitological infection rates in cattle samples maintained a weak relationship with entomological inoculation rates probably because of the limited time scale of sample collection. Trypanosome risk in Shimba Hills is high in locations close to the wildlife reserve and driven by G. pallidipes infectious bites on cattle. Therefore, trypanosome vector control programmes in the area should be designed to reduce G. pallidipes abundance and tailored to target sites close to the wildlife reserve.

Perspectives on Odor-Based Control of Tsetse Flies in Africa.

Tsetse-transmitted trypanosomiases are among the most neglected tropical diseases in sub-Sahara Africa. Although all tsetse species are susceptible to trypanosome infections, their differential attraction/feeding preferences for different wildlife, domestic animals, and/or humans constitute critical determinants of trypanosomes species they predominantly transmit. Artificial bait technologies, based on long-range tsetse olfactory responses to natural cues emitted by preferred hosts and blends of synthetic versions that mimic these cues, have successfully been applied in attractant-odor-based (“pull” tactic) reduction of field populations of some tsetse species. Olfactory attribute associated with active avoidance of tsetse-refractory non-hosts has similarly been exploited in design of repellent-odor-based (“push” tactic) protection of livestock. These tactics have opened possibility of spatially strategic deployment of the two sets of odor baits in “push-pull” tactics. Possibility of developing blends with enhanced attraction and repellence compared with those associated with savannah tsetse fly hosts and non-hosts, respectively, have been explored, where structure activity and blends of different components generated two novel blends. The studies evaluated structure activity and blends of different components. One based on attractive constituents associated with buffalo (Syncerus caffer) comprised of ε-nonalactone, nonanoic acid, 2-nonanone (in 1:3:2 proportion) delivered together with acetone, which showed significantly better attractancy on savannah tsetse fly than the standard blend comprised of 3-propylphenol, octenol, p-cresol, and acetone (POCA). The other blend comprised of δ-nonalactone, heptanoic acid, 4-methylguaiacol and geranylacetone (in 6:4:2:1 proportion) was significantly more repellent than previously characterized blend based on tsetse fly refractory waterbuck (Kobus defassa) constituents (δ-octalactone, pentanoic acid, guaiacol and geranylacetone). So far, no effective attractants or repellents of riverine tsetse fly species have been characterized. Optimized attractant and repellent blends for savannah tsetse flies lay down useful groundwork for future development of the “push-pull” deployment tactic for area-wide control of tsetse flies. Better understanding of the physiological, cellular, and molecular basis of response in the tsetse fly to odors can potentially augment the current tsetse fly-control interventions.

Xenomonitoring of trypanosome infectivity in Tse tse flies from Kagarko Local Government Area, Kaduna State Nigeria

Trypanosomiasis is a re-emerging neglected tropical disease, afflicting domestic, wild animals and humans. In this study, the tsetse fly infectivity and species composition of Trypanosoma species were determined in Kagarko Local Government Area, Kaduna State Nigeria using entomological protocol. The tsetse flies were sampled using standard biconical traps and the flies collected were dissected for trypanosome using microscopy as well as expression of tsetse antigen 5 (TAg 5) using molecular tools. Forty seven (47) tsetse flies were collected over a period of four months, of which 13 (34.21%) were infected with Trypanosoma spp. The species of trypanosome identified include T. vivax 7(53.85%), T. congolense 5(38.46%) and T. brucei 1(7.69%), while expression of TAg 5 was observed in four of the examined flies. The presence of the flies with trypanosome of veterinary and medical importance demonstrate the risk of transmission of trypanosomiasis in the study area.

Tsetse Flies: Comparative Morphometric Information from Traits Collected on Wings and Pupae

Tsetse flies (Diptera: Glossinidae) are pupiparous insects that transmit African trypanosomes to humans and animals. Using three species of tsetse flies belonging to two subgenera, Nemorhina and Glossina, our data provide a first answer to the following question: do the juvenile stages of tsetse flies provide metric traits allowing reliable species and/or sex identification? We compared three species using their wings and their pupae: Glossina palpalis gambiensis, Glossina tachinoides and Glossina morsitans submorsitans. To separate size and shape information, 11 landmarks per wing were submitted to the Generalised Procrustes Analysis (GPA). The contour of the pupae was processed by elliptic Fourier analysis (EFA).A validated reclassification method was used to compare the taxonomic power of each organ, as well as its utility for sex recognition. The contour of the pupae did not provide a satisfactory species signal (75 %), relative to the one obtained from wing landmarks (99 %). However, the overall pupae size was much more satisfactory than the wings size (93 % versus 57 %). In spite of a lower taxonomic signal, the morphometric variation observed on pupae was in agreement with known phylogenetic relationships. Contrary to the wings, sexual dimorphism was much reduced in pupae and not strong enough to allow any reliable sex identification.

Viviparity and habitat restrictions may influence the evolution of male reproductive genes in tsetse fly (Glossina) species

BackgroundGlossina species (tsetse flies), the sole vectors of African trypanosomes, maintained along their long evolutionary history a unique reproductive strategy, adenotrophic viviparity. Viviparity reduces their reproductive rate and, as such, imposes strong selective pressures on males for reproductive success. These species live in sub-Saharan Africa, where the distributions of the main sub-genera Fusca, Morsitans, and Palpalis are restricted to forest, savannah, and riverine habitats, respectively. Here we aim at identifying the evolutionary patterns of the male reproductive genes of six species belonging to these three main sub-genera. We then interpreted the different patterns we found across the species in the light of viviparity and the specific habitat restrictions, which are known to shape reproductive behavior.ResultsWe used a comparative genomic approach to build consensus evolutionary trees that portray the selective pressure acting on the male reproductive genes in these lineages. Such trees reflect the long and divergent demographic history that led to an allopatric distribution of the Fusca, Morsitans, and Palpalis species groups. A dataset of over 1700 male reproductive genes remained conserved over the long evolutionary time scale (estimated at 26.7 million years) across the genomes of the six species. We suggest that this conservation may result from strong functional selective pressure on the male imposed by viviparity. It is noteworthy that more than half of these conserved genes are novel sequences that are unique to the Glossina genus and are candidates for selection in the different lineages.ConclusionsTsetse flies represent a model to interpret the evolution and differentiation of male reproductive biology under different, but complementary, perspectives. In the light of viviparity, we must take into account that these genes are constrained by a post-fertilization arena for genomic conflicts created by viviparity and absent in ovipositing species. This constraint implies a continuous antagonistic co-evolution between the parental genomes, thus accelerating inter-population post-zygotic isolation and, ultimately, favoring speciation. Ecological restrictions that affect reproductive behavior may further shape such antagonistic co-evolution.

Diversity of tsetse flies and trypanosome species circulating in the area of Lake Iro in southeastern Chad

BackgroundAfrican trypanosomiases are vector-borne diseases that affect humans and livestock in sub-Saharan Africa. Although data have been collected on tsetse fauna as well as trypanosome infections in tsetse flies and mammals in foci of sleeping sickness in Chad, the situation of tsetse fly-transmitted trypanosomes remains unknown in several tsetse-infested areas of Chad. This study was designed to fill this epidemiological knowledge gap by determining the tsetse fauna as well as the trypanosomes infecting tsetse flies in the area of Lake Iro in southeastern Chad.MethodsTsetse flies were trapped along the Salamat River using biconical traps. The proboscis and tsetse body were removed from each fly. DNA was extracted from the proboscis using proteinase K and phosphate buffer and from the tsetse body using Chelex 5%. Tsetse flies were identified by amplifying and sequencing the cytochrome c oxydase I gene of each tsetse fly. Trypanosome species were detected by amplifying and sequencing the internal transcribed spacer 1 of infecting trypanosomes.ResultsA total of 617 tsetse flies were trapped; the apparent density of flies per trap per day was 2. 6. Of the trapped flies, 359 were randomly selected for the molecular identification and for the detection of infecting trypanosomes. Glossina morsitans submorsitans (96.1%) was the dominant tsetse fly species followed by G. fuscipes fuscipes (3.1%) and G. tachinoides (0.8%). Four trypanosome species, including Trypanosoma vivax, T. simiae, T. godfreyi and T. congolense savannah, were detected. Both single infection (56.7%) and mixed infections of trypanosomes (4.6%) were detected in G. m. submorsitans. The single infection included T. simiae (20.5%), T. congolense savannah (16.43%), T. vivax (11.7%) and T. godfreyi (9.8%). The trypanosome infection rate was 61.4% in G. m. submorsitans, 72.7% in G. f. fuscipes and 66.6% in G. tachinoides. Trypanosome infections were more prevalent in tsetse bodies (40.6%) than in the proboscis (16.3%).ConclusionThis study revealed the presence of different tsetse species and a diversity of trypanosomes pathogenic to livestock in the area of Lake Iro. The results highlight the risks and constraints that animal African trypanosomiasis pose to livestock breeding and the importance of assessing trypanosome infections in livestock in this area.Graphical

Characterization of a composite with enhanced attraction to savannah tsetse flies from constituents or analogues of tsetse refractory waterbuck (Kobus defassa) body odor.

Savannah tsetse flies avoid flying toward tsetse fly-refractory waterbuck (Kobus defassa) mediated by a repellent blend of volatile compounds in their body odor comprised of δ-octalactone, geranyl acetone, phenols (guaiacol and carvacrol), and homologues of carboxylic acids (C5-C10) and 2-alkanones (C8-C13). However, although the blends of carboxylic acids and that of 2-alkanones contributed incrementally to the repellency of the waterbuck odor to savannah tsetse flies, some waterbuck constituents (particularly, nonanoic acid and 2-nonanone) showed significant attractive properties. In another study, increasing the ring size of δ-octalactone from six to seven membered ring changed the activity of the resulting molecule (ε-nonalactone) on the savannah tsetse flies from repellency to attraction. In the present study, we first compared the effect of blending ε-nonalactone, nonanoic acid and 2-nonanone in 1:1 binary and 1:1:1 ternary combination on responses of Glossina pallidipes and Glossina morsitans morsitans tsetse flies in a two-choice wind tunnel. The compounds showed clear synergistic effects in the blends, with the ternary blend demonstrating higher attraction than the binary blends and individual compounds. Our follow up laboratory comparisons of tsetse fly responses to ternary combinations with different relative proportions of the three components showed that the blend in 1:3:2 proportion was most attractive relative to fermented cow urine (FCU) to both tsetse species. In our field experiments at Shimba Hills game reserve in Kenya, where G. pallidipes are dominant, the pattern of tsetse catches we obtained with different proportions of the three compounds were similar to those we observed in the laboratory. Interestingly, the three-component blend in 1:3:2 proportion when released at optimized rate of 13.71mg/h was 235% more attractive to G. pallidipes than a combination of POCA (3-n-Propylphenol, 1-Octen-3-ol, 4-Cresol, and Acetone) and fermented cattle urine (FCU). This constitutes a novel finding with potential for downstream deployment in bait technologies for more effective control of G. pallidipes, G. m. morsitans, and perhaps other savannah tsetse fly species, in 'pull' and 'pull-push' tactics.

Potential impacts of climate change on geographical distribution of three primary vectors of African Trypanosomiasis in Tanzania's Maasai Steppe: G. m. morsitans, G. pallidipes and G. swynnertoni.

In the Maasai Steppe, public health and economy are threatened by African Trypanosomiasis, a debilitating and fatal disease to livestock (African Animal Trypanosomiasis -AAT) and humans (Human African Trypanosomiasis-HAT), if not treated. The tsetse fly is the primary vector for both HAT and AAT and climate is an important predictor of their occurrence and the parasites they carry. While understanding tsetse fly distribution is essential for informing vector and disease control strategies, existing distribution maps are old and were based on coarse spatial resolution data, consequently, inaccurately representing vector and disease dynamics necessary to design and implement fit-for-purpose mitigation strategies. Also, the assertion that climate change is altering tsetse fly distribution in Tanzania lacks empirical evidence. Despite tsetse flies posing public health risks and economic hardship, no study has modelled their distributions at a scale needed for local planning. This study used MaxEnt species distribution modelling (SDM) and ecological niche modeling tools to predict potential distribution of three tsetse fly species in Tanzania's Maasai Steppe from current climate information, and project their distributions to midcentury climatic conditions under representative concentration pathways (RCP) 4.5 scenarios. Current climate results predicted that G. m. morsitans, G. pallidipes and G swynnertoni cover 19,225 km2, 7,113 km2 and 32,335 km2 and future prediction indicated that by the year 2050, the habitable area may decrease by up to 23.13%, 12.9% and 22.8% of current habitable area, respectively. This information can serve as a useful predictor of potential HAT and AAT hotspots and inform surveillance strategies. Distribution maps generated by this study can be useful in guiding tsetse fly control managers, and health, livestock and wildlife officers when setting surveys and surveillance programs. The maps can also inform protected area managers of potential encroachment into the protected areas (PAs) due to shrinkage of tsetse fly habitats outside PAs.