Glossina Morsitans Research Articles

Species richness and abundance of wild tsetse flies collected from selected human-wildlife-livestock interface in Tanzania

The successful control of tsetse flies largely depends on understanding of the species available and abundance. This study assessed the species richness, abundance and apparent density of wild collected tsetse flies from selected human-wildlife-livestock interface in Tanzania. Seasonal trapping using baited NZI, Pyramidal and Biconical traps was done across selected wards. Traps were set at 200 m apart, emptied after every 24 h then rotated to the next sites after 72 h. Collected flies were identified morphologically and letter confirmed using the Polymerase Chain Reaction (PCR). Only two Glossina species; Glossina pallidipes (n = 371; 47.32 %) and Glossina morsitans morsitans (n = 413; 52.68 %) were identified. Among them, 96 flies (80 Female, 16 Male) were blood fed; 57(48 Female and 9 Male) G. pallidipes and 39(32 Female and 7 Male) G.m. morsitans. Tsetse fly abundance varied across wards (χ2 = 4.597, df = 1, p = 0.032), villages (χ2 = 9.491, df = 3, p = 0.023), habitats (χ2 = 17.239, df = 2, p < 0.001), months (χ2 = 13.507, df = 3, p = 0.004) and deployed traps (χ2 = 6.348, df = 2, p = 0.04). About 78.82 % of the total catch occurred in Kisaki ward (n = 618; p < 0.001) and 21.17 % (n = 166; p = 0.032) in Bwakila chini. Similarly, 62.37 % of the catch occurred in Mbojoge village. NZI traps (n = 422; 54 %; 4.98 FTD) were most successful traps. Moreover, 78.06 % of the catch occurred in bushed grassland habitat (n = 612; 55.41 FTD) while 5.48 % in farmland (n = 43; 7.17 FTD). This study recommends NZI and Pyramidal traps for tsetse flies control at the interface and proposes wet season as appropriate time for successful trapping of the flies. Finally, it attracts a need for assessing tsetse flies' blood meal sources and the infection status to establish the prevalence to inform existing trypanosome control programs.

Phenotypic divergence of Glossina morsitans (Diptera: Glossinidae) populations in Zambia: Application of landmark-based wing geometric morphometrics to discriminate population-level variation.

An important consequence of the discontinuous distribution of insect populations within their geographic range is phenotypic divergence. Detection of this divergence can be challenging when it occurs through subtle shifts in morphological traits with complex geometries, such as insect wing venation. Here, we used landmark-based wing geometric morphometrics to investigate the population-level phenotypic variation of the two subspecies of Glossina morsitans, G. m. centralis Machado and G. m. morsitans Westwood that occur in Zambia. Twelve homologous landmarks digitised on the right wings of 720 specimens collected from four and five sites (80 per site with 1:1 sex ratio) within the G. m. centralis and G. m. morsitans range respectively, were subjected to generalised Procrustes analysis to obtain wing centroid size (CS) and wing shape variables. Linear permutation models and redundancy analysis were then used to compare CS and wing shape between male and female G. morsitans, the two subspecies G. m. centralis and G. m. morsitans, the sexes of each subspecies and between sample locations within each subspecies range, respectively. Significant differences in CS and wing shape were observed between G. morsitans sexes, subspecies and sample locations within each subspecies range. A neighbour-joining cladogram derived from the analysis of Procrustes distances showed that tsetse within each subspecies range were highly divergent. We conclude that G. morsitans populations in Zambia exhibit significant population-level variation in fly size and wing shape which suggests high levels of population structuring. The main drivers of this structuring could be random genetic drift in G. m. centralis demes and local adaptation to environmental conditions in G. m. morsitans populations. We therefore recommend molecular studies to estimate the levels of gene flow between these populations and identify possible barriers to genetic flow.

Optical fibre based artificial compound eyes for direct static imaging and ultrafast motion detection

Natural selection has driven arthropods to evolve fantastic natural compound eyes (NCEs) with a unique anatomical structure, providing a promising blueprint for artificial compound eyes (ACEs) to achieve static and dynamic perceptions in complex environments. Specifically, each NCE utilises an array of ommatidia, the imaging units, distributed on a curved surface to enable abundant merits. This has inspired the development of many ACEs using various microlens arrays, but the reported ACEs have limited performances in static imaging and motion detection. Particularly, it is challenging to mimic the apposition modality to effectively transmit light rays collected by many microlenses on a curved surface to a flat imaging sensor chip while preserving their spatial relationships without interference. In this study, we integrate 271 lensed polymer optical fibres into a dome-like structure to faithfully mimic the structure of NCE. Our ACE has several parameters comparable to the NCEs: 271 ommatidia versus 272 for bark beetles, and 180o field of view (FOV) versus 150–180o FOV for most arthropods. In addition, our ACE outperforms the typical NCEs by ~100 times in dynamic response: 31.3 kHz versus 205 Hz for Glossina morsitans. Compared with other reported ACEs, our ACE enables real-time, 180o panoramic direct imaging and depth estimation within its nearly infinite depth of field. Moreover, our ACE can respond to an angular motion up to 5.6×106 deg/s with the ability to identify translation and rotation, making it suitable for applications to capture high-speed objects, such as surveillance, unmanned aerial/ground vehicles, and virtual reality.

Killing tsetse and/or saving wildlife? A multispecies assemblage in colonial Zambia (1895–1959)

This article investigates the problem of the tsetse fly and the trypanosomiasis disease it conveys as a transforming multispecies assemblage in colonial Zambia from the late nineteenth century until 1959. Based on archival research, it analyses the tsetse fly (Glossina morsitans) as a moving target; not only a mobile and elusive insect but also a moving field of knowledge bringing multiple stakeholders into dialogue. It shows that tsetse control and wildlife conservation emerged together in colonial Zambia, in conflicting but also synergising ways, and that the association of large mammals to G. morsitans laid the ground for their classification as killable or preservable species. In the crossed influence of diverse regional colonial expertise, the article finds that the complex multispecies relations between the tsetse fly, the trypanosomes, wildlife, vegetation, humans and cattle, mediated and enacted by colonial experts and others, shaped institutions, policies and landscapes.

Establishment and partial characterisation of a new cell line derived from adult tissues of the tsetse fly Glossina morsitans morsitans

BackgroundInsect cell lines play a vital role in many aspects of research on disease vectors and agricultural pests. The tsetse fly Glossina morsitans morsitans is an important vector of salivarian trypanosomes in sub-Saharan Africa and, as such, is a major constraint on human health and agricultural development in the region.MethodsHere, we report establishment and partial characterisation of a cell line, GMA/LULS61, derived from tissues of adult female G. m. morsitans. GMA/LULS61 cells, grown at 28 °C in L-15 (Leibovitz) medium supplemented with foetal bovine serum and tryptose phosphate broth, have been taken through 23 passages to date and can be split 1:1 at 2-week intervals. Karyotyping at passage 17 revealed a predominantly haploid chromosome complement. Species origin and absence of contaminating bacteria were confirmed by PCR amplification and sequencing of fragments of the COI gene and pan-bacterial 16S rRNA gene respectively. However, PCR screening of RNA extracted from GMA/LULS61 cells confirmed presence of the recently described Glossina morsitans morsitans iflavirus and Glossina morsitans morsitans negevirus, but absence of Glossina pallipides salivary gland hypertrophy virus. GMA/LULS61 cells supported infection and growth of 6/7 different insect-derived strains of the intracellular bacterial symbiont Wolbachia.ConclusionsThe GMA/LULS61 cell line has potential for application in a variety of studies investigating the biology of G. m. morsitans and its associated pathogenic and symbiotic microorganisms.Graphical

Updating the northern tsetse distribution limit in Chad in the context of global change

Tsetse flies (Diptera: Glossinidae) are obligate bloodfeeders that occur exclusively in Sub-Saharan Africa, where they are the vectors of trypanosomes causing HAT (human African trypanosomiasis) and AAT (African animal trypanosomiasis). In Chad, tsetse flies occur only in the most southern part of the country because of its favorable bioclimatic conditions. However, despite the importance of HAT and AAT in this country, very little is known about the current tsetse distribution, in particular its northern limit, which is of key importance for the surveillance of these diseases. A total of 217 biconical traps were deployed in 2021 and 2022 from the West to the East around the formerly known northern limit, resulting in 1,024 tsetse caught belonging to three different taxa: Glossina morsitans submorsitans (57%), G. tachinoides (39%) and G. fuscipes fuscipes (4%). In addition to the information gathered on the presence/absence of each tsetse taxon, we show a strong North-South shift of the northen tsetse distribution limit as compared to the previous works from 1966 to 1996, and a growing spatial fragmentation in more and more discrete pockets of tsetse presence. This North-South shift of the northern tsetse distribution limit in Chad is the likely consequence of the combined effect of severe draughts that affected the country, and increasing human pressure on land. This update of the tsetse northern limit will be of help to the national programmes in charge of HAT and AAT.

Host Choice and Feeding Behaviours of Glossina morsitans Offspring Whose Parents Were Fed on Different Host Species

The success of any tsetse control program depends on the knowledge of their behaviour. This study assessed the host choice and feeding behaviours of Glossina morsitans siblings whose parents were bloodfed on rabbits, guinea pigs, rodents, and squirrels. Each individual host was placed in a screen cage, which allowed flies to enter through openings on each side. The groups of flies (20 per replicate), which were colour-marked differently based on their parents’ blood meal hosts, were released from the centre of large semi-field cage. The released flies were aspirated after 24 h and then sorted based on their location, feeding status, and parents’ blood meal. A total of 213 flies (72.95% of those recovered) were attracted to the hosts. The numbers of flies attracted to different hosts varied significantly (χ24 = 33.685, p = 0.0001): rodents (n = 80, p = 0.006), rabbits (n = 59, p = 0.331), guinea pigs (n = 49, p = 0.057), and squirrels (n = 25, p = 0.005). The numbers of flies attracted to their parent’s blood meal source varied significantly (χ212 = 56.476, p &lt; 0.001): rabbits (n = 35, 59.32%, p &lt; 0.001), rodents (n = 25, 31.25%, p = 0.043), and guinea pigs (n = 19, 38.78%, p = 0.45). But only 39 flies (18.31% of the total attracted) bloodfed on the hosts, including guinea pigs (n = 10, 25.64%), rodents (n = 23, 58.97%), rabbits (n = 6, 15.38%), and squirrels (n = 0, 0.0%). There was significant variation in the number of flies that fed successively across hosts (χ24 = 49.478, p &lt; 0.001). The findings from this study confirm the presence of differential attractiveness of the hosts to flies and the so-called “Hopkins host selection principle” or “pre-imaginal conditioning”. Therefore, the study attracts the need for detailed investigation on the influence of blood meal sources on tsetse fly siblings’ behaviours across filial generations using small mammals or other large mammal species.

Advancing age grading techniques for Glossina morsitans morsitans, vectors of African trypanosomiasis, through mid-infrared spectroscopy and machine learning.

Tsetse are the insects responsible for transmitting African trypanosomes, which cause sleeping sickness in humans and animal trypanosomiasis in wildlife and livestock. Knowing the age of these flies is important when assessing the effectiveness of vector control programs and modelling disease risk. Current methods to assess fly age are, however, labour-intensive, slow, and often inaccurate as skilled personnel are in short supply. Mid-infrared spectroscopy (MIRS), a fast and cost-effective tool to accurately estimate several biological traits of insects, offers a promising alternative. This is achieved by characterising the biochemical composition of the insect cuticle using infrared light coupled with machine-learning (ML) algorithms to estimate the traits of interest. We tested the performance of MIRS in estimating tsetse sex and age for the first-time using spectra obtained from their cuticle. We used 541 insectary-reared Glossina m. morsitans of two different age groups for males (5 and 7 weeks) and three age groups for females (3 days, 5 weeks, and 7 weeks). Spectra were collected from the head, thorax, and abdomen of each sample. ML models differentiated between male and female flies with a 96% accuracy and predicted the age group with 94% and 87% accuracy for males and females, respectively. The key infrared regions important for discriminating sex and age classification were characteristic of lipid and protein content. Our results support the use of MIRS as a rapid and accurate way to identify tsetse sex and age with minimal pre-processing. Further validation using wild-caught tsetse could pave the way for this technique to be implemented as a routine surveillance tool in vector control programmes.

Diversity of Glossinidae (Diptera) species in The Gambia in relation to vegetation.

Glossina species are known to transmit African Trypanosomiasis, one of the most important infectious diseases for both livestock and humans in sub-Saharan Africa. Therefore, the aim of this study was to characterize trapped Glossina spp. from The Gambia using morphological and molecular techniques in relation to the vegetation cover types. A line transect survey was carried out in all the administrative regions of The Gambia. Tsetse fly trapping was carried out for 14 days during each season using line transect. A total of 220 Glossina spp. specimens (117 F and 103 M) were captured, and DNA was extracted from the legs of 100 randomly selected Glossina spp. Further, DNA samples were tested by a conventional PCR assay. A total of 135/220 (61%; 95% CI: 54.6-67.8%) and 85/220 (39%; 95% CI: 32.2-45.4%) flies were identified as Glossina morsitans submorsitans and Glossina palpalis gambiensis, respectively, with most caught during wet season (53.6%) and more females (53.2%) than males. Results of the morphological identification agreed with those of molecular identification. The type of vegetation cover significantly influenced the caught of tsetse flies. Animals and humans at the various trapping sites are at risk of being bitten by tsetse flies.

Rearing of Glossina morsitans morsitans tsetse flies for the sterile insect technique: evaluating the impact of irradiation and transportation during early and late-stage pupal development on the quality of emerging adults.

Human African trypanosomiasis (HAT) and African animal trypanosomosis (AAT) are devastating diseases spread by tsetse flies (Glossina spp.), affecting humans and livestock, respectively. Current efforts to manage these diseases by eliminating the vector through the sterile insect technique (SIT) require transportation of irradiated late-stage tsetse pupae under chilling, which has been reported to reduce the biological quality of emerged flies. We therefore evaluated the impact of irradiation and transportation (including vibration and shock) on pupae at early-stage development (22 days of age) under ambient temperature and compared it to that on pupae at the late-stage development (29 days of age) under chilling, the current practice for tsetse in SIT programs. The quality of flies emerging from these transported pupae was assessed by their emergence rates, flight propensity, mating ability, insemination rates and survival rates (over ca. 100 days, and after specified shorter periods). Generally, flies emerging from the 22-day-old pupae had significantly (p<0.05) higher values for the tested quality parameters, as compared to those emerging from 29-day-old pupae. Irradiation, transportation and the combination thereof significantly (p<0.05) reduced all the tested quality parameters as compared with the untreated control within the 22-day-old pupae group. Further, vibration had a significant negative effect on the quality of flies, notwithstanding the age of the pupae. Irradiation and transportation of pupae at 22 days of age resulted in a higher proportion of flies of good biological quality as compared to those of 29 days of age, and hence may be considered for future SIT programs.

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.

Spatial distribution of Glossina morsitans (Diptera: Glossinidae) in Zambia: A vehicle-mounted sticky trap survey and Maxent species distribution model.

Tsetse-transmitted African trypanosomiasis is a debilitating and fatal disease of humans and livestock if left untreated. While knowledge of the spatial distribution patterns of tsetse is essential for the development of risk-based vector control strategies, existing distribution maps in Zambia are more than 40 years old and were based on coarse spatial resolution data. The recently developed vehicle-mounted sticky trap (VST) provides an alternative sampling device to aid in updating existing distribution maps but has not been applied outside an experimental setting and is limited to motorable tracks. Therefore, the objective of the present study was to demonstrate the effectiveness of utilizing the VST for area-wide surveys of Glossina morsitans and to use the occurrence records to predict its spatial distribution in Zambia under current environmental conditions using Maxent. Two-sided all-blue VST baited with butanone and 1-octen-3-ol was used to survey 692 and 1020 km of transect routes in G. m. centralis Machado and G. m. morsitans Westwood previously published distribution in Zambia. Maxent species distribution technique was used to predict the potential distribution of the two subspecies using current climatic and environmental data which was then compared to the historical distribution. A total of 15,602 tsetse were captured with G. m. morsitans (58%) being the most abundant. G. m. centralis and G. pallidipes Austin represented 39 and 2% of the catch respectively, and G. brevipalpis Newstead was also detected. The predicted potential distribution for G. m. centralis was 80,863 km2 while that of G. m. morsitans was 70,490 km2 representing a 47 and 29% reduction compared to their historical distributions, respectively. The VST is effective for sampling G. morsitans outside experimental settings and is recommended for use as an additional tsetse survey tool. The spatial distribution of G. morsitans in Zambia has reduced by 101,051 km2 due to temperature and land cover changes.

Understanding Tsetse Fly (Glossina morsitans) Behavior through its Genome

&lt;em&gt;Glossina morsitans (G. morsitans), &lt;/em&gt;commonly known as tsetse fly, have caused public health concerns throughout the years. &lt;em&gt;G. morsitans&lt;/em&gt; is the vector for &lt;em&gt;Trypanosoma brucei &lt;/em&gt;(&lt;em&gt;T. brucei&lt;/em&gt;), the parasite responsible for causing the deadly African sleeping disease (African trypanosomiasis). Researchers have searched for ways to contain this disease, but to little avail. Fortunately, new advances in sequencing methods have given researchers a new opportunity to win the war against the disease. The whole-genome sequence of &lt;em&gt;G. morsitans &lt;/em&gt;provides essential data regarding involved genes that transmits &lt;em&gt;T. brucei&lt;/em&gt; to humans. Information about those unique genes facilitates researchers to create new methods to prevent &lt;em&gt;G. morsitans&lt;/em&gt; from becoming the vector of &lt;em&gt;T. brucei&lt;/em&gt;, enabling the containment of this disease. With this, we review the unique genes in the &lt;em&gt;G. morsitans&lt;/em&gt; genome, such as those that contribute to blood-feeding ability, establish a relationship with symbionts, and &lt;em&gt;G. morsitans &lt;/em&gt;unique sensory genes, with an expectation that it would enhance our knowledge of &lt;em&gt;G. morsitans&lt;/em&gt; as the vector for parasites causing African trypanosomiasis.

Lipid metabolism dysfunction following symbiont elimination is linked to altered Kennedy pathway homeostasis

Lipid metabolism is critical for insect reproduction, especially for species that invest heavily in the early developmental stages of their offspring. The role of symbiotic bacteria during this process is understudied but likely essential. We examined the role of lipid metabolism during the interaction between the viviparous tsetse fly (Glossina morsitans morsitans) and its obligate endosymbiotic bacteria (Wigglesworthia glossinidia) during tsetse pregnancy. We observed increased CTP:phosphocholine cytidylyltransferase (cct1) expression during pregnancy, which is critical for phosphatidylcholine biosynthesis in the Kennedy pathway. Experimental removal of Wigglesworthia impaired lipid metabolism via disruption of the Kennedy pathway, yielding obese mothers whose developing progeny starve. Functional validation via experimental cct1 suppression revealed a phenotype similar to females lacking obligate Wigglesworthia symbionts. These results indicate that, in Glossina, symbiont-derived factors, likely B vitamins, are critical for the proper function of both lipid biosynthesis and lipolysis to maintain tsetse fly fecundity.

Improved estimates of abortion rates in tsetse (Glossina spp.).

Abortion rates were assessed among 170, 846 tsetse (154,228 Glossina pallidipes and 19,618 Glossina morsitans morsitans) sampled in Zimbabwe in 1988-1999. The study produced improved estimates of abortion rates and how these varied with fly age and size and temperatures experienced during pregnancy. An abortion was diagnosed if the uterus was empty and the largest oocyte <0.82 of the expected mature length. Abortion rates for G. pallidipes and G. m. morsitans were 0.64% (95% ci: 0.59-0.69) and 0.83% (0.62-1.10) for trapped flies and 2.03% (1.77-2.31) and 1.55% (1.20-1.98) for flies from artificial refuges. Abortion rates increased with increasing temperature and decreased with increasing wing length and wing fray. Contrary to laboratory findings, abortion rates did not increase in the oldest flies. Percentages of tsetse with empty uteri, regardless of abortion status, were significantly higher than estimated abortion percentages. For tsetse from traps, 4.01% (95% ci: 3.90-4.13) of G. pallidipes and 2.52% (2.14-2.95) of G. m. morsitans had empty uteri; for flies from artificial refuges, the percentages were 12.69% (12.07-13.34) and 14.90% (13.82-16.02), respectively. Abortion losses are small relative to losses at all other stages of life.

Trypanosomes infection, endosymbionts, and host preferences in tsetse flies (Glossina spp.) collected from Akagera park region, Rwanda: A correlational xenomonitoring study

Akagera National Park and its surroundings are home to tsetse flies and a number of their mammalian hosts in Rwanda. A One-health approach is being used in the control and surveillance of both animal and human trypanosomosis in Rwanda. Determination of the infection level in tsetse flies, species of trypanosomes circulating in vectors, the source of tsetse blood meal and endosymbionts is crucial in understanding the epidemiology of the disease in animals and humans in the region.Tsetse flies (n = 1101), comprising Glossina pallidipes (n = 771) and Glossina morsitans centralis (n = 330) were collected from Akagera park and surrounding areas between May 2018 and June 2019. The flies were screened for trypanosomes, vertebrate host DNA to identify sources of blood meal, and endosymbionts by PCR - High Resolution Melting analysis and amplicon sequencing. The feeding frequency and the feeding indices (selection index - W) were calculated to identify the preferred hosts. An overall trypanosome infection rate of 13.9% in the fly's Head and Proboscis (HP) and 24.3% in the Thorax and Abdomen (TA) were found. Eight trypanosome species were identified in the tsetse fly HP and TA, namely: Trypanosoma (T.) brucei brucei, T. congolense Kilifi, T. congolense savannah, T. vivax, T. simiae, T. evansi, T. godfreyi, T. grayi and T. theileri. We found no evidence of human-infective T. brucei rhodesiense. We also identified eighteen species of vertebrate hosts that tsetse flies fed on, and the most frequent one was the buffalo (Syncerus caffer) (36.5%). The frequently detected host by selection index was the rhinoceros (Diceros bicornis) (W = 16.2). Most trypanosome infections in tsetse flies were associated with the buffalo blood meal. The prevalence of tsetse endosymbionts Sodalis and Wolbachia was 2.8% and 4.8%, respectively. No Spiroplasma and Salivary Gland Hypertrophy Virus were detected. These findings implicate the buffaloes as the important reservoirs of tsetse-transmitted trypanosomes in the area. This contributes to predicting the main cryptic reservoirs and therefore guiding the effective control of the disease. The study findings provide the key scientific information that supports the current One Health collaboration in the control and surveillance of tsetse-transmitted trypanosomosis in Rwanda.

Tsetse flies (Glossina morsitans morsitans) choose birthing sites guided by substrate cues with no evidence for a role of pheromones.

Tsetse flies significantly impact public health and economic development in sub-Saharan African countries by transmitting the fatal disease African trypanosomiasis. Unusually, instead of laying eggs, tsetse birth a single larva that immediately burrows into the soil to pupate. Where the female chooses to larviposit is, therefore, crucial for offspring survival. Previous laboratory studies suggested that a putative larval pheromone, n-pentadecane, attracts gravid female Glossina morsitans morsitans to appropriate larviposition sites. However, this attraction could not be reproduced in field experiments. Here, we resolve this disparity by designing naturalistic laboratory experiments that closely mimic the physical characteristics found in the wild. We show that gravid G. m. morsitans were neither attracted to the putative pheromone nor, interestingly, to pupae placed in the soil. By contrast, females appear to choose larviposition sites based on environmental substrate cues. We conclude that, among the many cues that likely contribute to larviposition choice in nature, substrate features are a main determinant, while we failed to find evidence for a role of pheromones.

Evaluating the Effect of Irradiation on the Densities of Two RNA Viruses in Glossina morsitans morsitans.

Tsetse flies are cyclic vectors of Trypanosoma parasites, which cause debilitating diseases in humans and animals. To decrease the disease burden, the number of flies is reduced using the sterile insect technique (SIT), where male flies are sterilized through irradiation and released into the field. This procedure requires the mass rearing of high-quality male flies able to compete with wild male flies for mating with wild females. Recently, two RNA viruses, an iflavirus and a negevirus, were discovered in mass-reared Glossina morsitans morsitans and named GmmIV and GmmNegeV, respectively. The aim of this study was to evaluate whether the densities of these viruses in tsetse flies are affected by the irradiation treatment. Therefore, we exposed tsetse pupae to various doses (0-150 Gy) of ionizing radiation, either in air (normoxia) or without air (hypoxia), for which oxygen was displaced by nitrogen. Pupae and/or emerging flies were collected immediately afterwards, and at three days post irradiation, virus densities were quantified through RT-qPCR. Generally, the results show that irradiation exposure had no significant impact on the densities of GmmIV and GmmNegeV, suggesting that the viruses are relatively radiation-resistant, even at higher doses. However, sampling over a longer period after irradiation would be needed to verify that densities of these insect viruses are not changed by the sterilisation treatment.

A volatile sex attractant of tsetse flies.

Tsetse flies transmit trypanosomes-parasites that cause devastating diseases in humans and livestock-across much of sub-Saharan Africa. Chemical communication through volatile pheromones is common among insects; however, it remains unknown if and how such chemical communication occurs in tsetse flies. We identified methyl palmitoleate (MPO), methyl oleate, and methyl palmitate as compounds that are produced by the tsetse fly Glossina morsitans and elicit strong behavioral responses. MPO evoked a behavioral response in male-but not virgin female-G. morsitans. G. morsitans males mounted females of another species, Glossina fuscipes, when they were treated with MPO. We further identified a subpopulation of olfactory neurons in G. morsitans that increase their firing rate in response to MPO and showed that infecting flies with African trypanosomes alters the flies' chemical profile and mating behavior. The identification of volatile attractants in tsetse flies may be useful for reducing disease spread.