Molecular evidence of Anaplasma spp. in blood-sucking flies from China and identification of a putative novel Anaplasma species.

Tabanids and stomoxes are important mechanical vectors for the transmission of pathogens. Although the agents they transmitted have been well studied, bacteria of the genus Anaplasma harbored by these flies have never been reported in China. In this study, 262 blood-sucking flies (128 Stomoxys calcitrans, 45 Tabanus birmanicus, 69 Tabanus hypomacros, and 20 Tabanus taiwanus) were collected from the Wuhan and Nanping cities of China. Anaplasma marginale, Anaplasma bovis, and Candidatus Anaplasma cinensis are detected in S. calcitrans from Wuhan City, with positive rates of 15.63%, 1.56%, and 7.81%, respectively. Out of our expectations, a putative novel Anaplasma species was identified in all three tabanid species (40.00% in T. birmanicus, 15.94% in T. hypomacros, and 10.00% in T. taiwanus) from Nanping City. The 16 S rRNA and groEL gene sequences have highest 99.37-99.75% and 91.46% identities to A. marginale, while the gltA gene sequences have highest 88.34% identity to Anaplasma centrale. In the phylogenetic trees, these strains form a distinct clade. Herein we name it "Candidatus Anaplasma nanpingensis". The present study shows the existence of multiple Anaplasma species in blood-sucking flies in China. This may be the first report that blood-sucking flies harbor Anaplasma in China.

Why don't horseflies land on zebras?

Stripes deter horseflies (tabanids) from landing on zebras and, while several mechanisms have been proposed, these hypotheses have yet to be tested satisfactorily. Here, we investigated three possible visual mechanisms that could impede successful tabanid landings (aliasing, contrast and polarization) but additionally explored pattern element size employing video footage of horseflies around differently patterned coats placed on domestic horses. We found that horseflies are averse to landing on highly but not on lightly contrasting stripes printed on horse coats. We could find no evidence for horseflies being attracted to coats that better reflected polarized light. Horseflies were somewhat less attracted to regular than to irregular check patterns, but this effect was not large enough to support the hypothesis of disrupting optic flow through aliasing. More likely it is due to attraction towards larger dark patches present in the irregular check patterns, an idea bolstered by comparing landings to the size of dark patterns present on the different coats. Our working hypothesis for the principal anti-parasite features of zebra pelage are that their stripes are sharply outlined and thin because these features specifically eliminate the occurrence of large monochrome dark patches that are highly attractive to horseflies at close distances.

New materials for improving catches of horseflies (Diptera: Tabanidae) in Nzi traps.

New materials in 85 configurations were tested relative to standard fabric Nzi traps to improve catches of tabanids based on artificial visual cues that mimic those used in nature for locating hosts or water. Colour-fast synthetic fabrics and photo-selective horticultural mesh were tested to facilitate insecticide impregnation and for improved durability. Many plastics were explored to document how flies react to polarized and visible vs. ultraviolet light. Lastly, sticky horizontal, linearly polarizing ground-level shiny plastic targets were tested for capturing water-seeking tabanids relative to traps and vertical fabric targets. Based on the capture of 281 786 female Tabanidae (Tabanus Linnaeus, Hybomitra Enderlein, Chrysops Meigen), four alternative formats provided higher catches and/or durability. They were substituting: (1) phthalogen turquoise for phthalogen blue, (2) solution-dyed fabrics for blue and black cotton, (3) clear/white horticultural mesh for mosquito netting, and (4) clear, reflective plastic for some or all netting. A Sunbrella/BioNet fabric trap (portable) and a blue-black Plexiglas®/netting trap (fixed applications) consistently performed best (1.5-3 × higher catches). Ground-level targets captured many male and gravid female Hybomitra but were ineffective for Tabanus. Traps and vertical fabric or transparent sticky targets captured mainly unfed females.

Efficiency of six different octenol-baited traps for collecting horseflies (Diptera: Tabanidae).

The efficiency and the potential species preference for six different horsefly trap types was studied. A total of 2401 horseflies were collected during this study. The most effective trap was the modified box trap with (27.9%), followed by the Nzi trap with (24.5%), Malaise trap (Townes design 1972) with (22.5)%, canopy trap with (17.9%), modified small canopy trap with (7.0%) and Malaise trap (Townes design 1962) with (0.2%). The Nzi and Malaise trap (Townes design 1972) did not differ significantly in the number of collected horseflies. The comparison of all other traps revealed significant differences in the number of collected horseflies. Tabanus bromius was the most abundant species with (58.6%) of the collected sample. The modified box trap was the most successful in collecting of T. bromius, while the Malaise trap (Townes design 1972) was the most successful in collecting the species Haematopota pluvialis, T. maculicornis and T. tergestinus. This study demonstrated that different trap designs influenced the quantity, species and taxonomic diversity of horseflies that were collected. Variables in the trap design included cloth type and colour and size and shape of trap. More detailed studies should be done to determine which factor or factors are most significant.

Characterization of tabanid flies (Diptera: Tabanidae) in South Africa and Zambia and detection of protozoan parasites they are harbouring.

Tabanids are haematophagous flies feeding on livestock and wildlife. In the absence of information on the relationship of tabanid flies and protozoan parasites in South Africa and Zambia, the current study was aimed at characterizing tabanid flies collected in these two countries as well as detecting protozoan parasites they are harbouring. A total of 527 tabanid flies were collected whereby 70·2% were from South Africa and 29·8% were from Zambia. Morphological analysis revealed a total of five different genera collected from the sampled areas namely: Ancala, Atylotus, Haematopota, Philoliche and Tabanus. DNA extracted from South African Tabanus par and Tabanus taeniola tested positive for the presence of Trypanosoma congolense (Savannah) and Trypanosoma theileri whilst one member from T. par was positive for Trypanosoma brucei species. DNA extracted from Zambian tabanid flies tested positive for the presence of Besnoitia species at 1·27% (2/157), Babesia bigemina 5·73% (9/157), Theileria parva 30·11% (30/157) and 9·82% (14/157) for Trypanosoma evansi. This study is the first to report on relationship of Babesia and Theileria parasites with tabanid flies. Further investigations are required to determine the role of tabanids in transmission of the detected protozoan parasites in livestock and wildlife in South Africa and Zambia.

Survey of Rickettsia species in hematophagous arthropods from endemic areas for Japanese spotted fever in China.

Japanese spotted fever (JSF) is caused by Rickettsia japonica, mainly vectored by hard ticks. However, whether R. japonica can be transmitted by other arthropods remains unknown. Moreover, it is of interest to investigate whether other Rickettsia species cause spotted fever in endemic areas. In this study, a survey of Rickettsia species was performed in hematophagous arthropods (mosquitoes, tabanids, and ticks) from endemic areas for JSF in Hubei Province, central China. The results showed that the diversity and prevalence of Rickettsia species in mosquitoes are low, suggesting that mosquitoes may not be the vector of zoonotic Rickettsia species. A novel Rickettsia species showed a high prevalence (16.31%, 23/141) in tabanids and was named "Candidatus Rickettsia tabanidii." It is closely related to Rickettsia from fleas and mosquitoes; however, its pathogenicity in humans needs further investigation. Five Rickettsia species were identified in ticks. Rickettsia japonica, the agent of JSF, was detected only in Haemaphysalis longicornis and Haemaphysalis hystricis, suggesting that they may be the major vectors of R. japonica. Notably, two novel species were identified in H. hystricis ticks, one belonging to the spotted fever group and the other potentially belonging to the ancestral group. The latter one named "Candidatus Rickettsia hubeiensis" may provide valuable insight into the evolutionary history of Rickettsia.

Horse Flies (Diptera: Tabanidae) in Mangrove Forests and Estuarine Floodplains on Marajó Island, Brazil.

Tabanids constitute an important group of hematophagous insects that can transmit zoonoses, but with studies on the ecological distribution of species still neglected in the Amazon. We evaluated the role of mangrove forests and estuarine floodplains located inside and outside a conservation unit (UC) on the coast of Marajó Island, Amazon River estuary, on the diversity and distribution of tabanids. Specifically, we studied whether the community of mangrove and estuarine floodplain tabanids located inside and outside the UC differ in abundance, richness, and species composition. We collected tabanids using a Malaise trap at 40 sampling points, resulting in 637 specimens distributed in 13 species and one morphotype, representing approximately 37% of the tabanid fauna ever recorded for the Marajó Island. There was no significant difference in the richness and composition of tabanids between the phytophysiognomies, but the abundance was significantly different, with greater abundance in the mangrove. The areas inside and around the UC had an influence on the tabanids, with the areas inside the UC having the highest number of specimens and species, also influencing the species composition. Two species are new records for the Marajó Island, bringing the number of species recorded to 38. Our results suggest that, along the Amazonian coast, mangroves and estuarine floodplains maintain part of the diversity of tabanids known for the Brazilian Amazon. Our data also indicate that the region's UC provides potentially important habitats for the maintenance of local tabanid populations.

First study on molecular detection of hemopathogens in tabanid flies (Diptera: Tabanidae) and cattle in Southern Thailand.

Background and Aim: Female tabanids play a key role in disease transmission as mechanical vectors for various hemopathogens, but only a limited number of studies have been conducted on them. This study aimed to investigate the occurrence of hemopathogens in tabanid flies compared to those found in nearby cattle hosts. Materials and Methods: Tabanids were collected using a Nzi trap for three consecutive days per month during the dry season (February-May 2021). Furthermore, blood samples were collected from 20 beef cattle (Bos taurus) raised in the same area where the flies were captured. Conventional polymerase chain reaction (PCR) was used to detect hemopathogenic DNA in flies and beef cattle. Results: In total, 279 female tabanids belonging to five species were collected: Tabanus megalops, Tabanus rubidus, Tabanus mesogaeus, Chrysops dispar, and Chrysops fuscomarginalis. Notably, T. megalops was the most abundant species, accounting for 89.2% of the flies collected (n = 249). PCR technique revealed that 76.6% of T. megalops carried at least one pathogen (Anaplasma, Ehrlichia, Babesia, or Theileria). In addition, all beef cattle had multiple hemopathogenic infections (Anaplasma marginale, Ehrlichia spp., Babesia bigemina, Babesia bovis, and Theileria spp.). Conclusion: Although T. megalops could carry many hemopathogens, it might not be an important vector due to the limited number of flies and parasitic load. Furthermore, T. megalops could be utilized to monitor the presence of hemopathogens in the study area, but not the disease occurrence in the individual host species. Knowing the presence of hemopathogens in flies could help manage the disease in this area.

The Use of "Tail-Pedometers" to Evaluate the Impact of Dipterans in Feeder Cattle.

Hematophagous flies are a pest for livestock; their direct impact reduces productivity, and they are vectors of parasites, bacteria and viruses. Their control using insecticides is inefficient and highly polluting. The validation of new control tools requires efficacy and cost-effectiveness evaluation. The quantification of hematophagous insects' impact in livestock is a challenging prerequisite. Tail flicks counts can reliably evaluate fly-burden; however, visual records are tedious and time-consuming. In the present study, automation of tail flick counts was made through the use of pedometers attached to the tail, in two groups of feeder cattle. Group A was kept in a pen under the protection of a mosquito net, and Group B was kept in an open-air pen. The fly density of Group B was evaluated using fly traps. The apparent density per trap ranged from 130 to 1700 in the study. The mean pedometer records per 24 h ranged from 957+/-58 bits in Group A to 11,138+/-705 bits in Group B. The night/day records observed in Group A (200/800 bits) were drastically increased in Group B (1000-4000/4000-14,000 bits) and variable along seasons. A very high correlation was observed between fly density and visual records or pedometer records (PR). Two-hour PRs proved to be a reliable predictive tool for fly density. Moreover, the pedometers revealed an unsuspected but significant nuisance of mosquitoes, which should be thoroughly investigated.

Tabanid-transmitted animal trypanosomiasis in Cameroon: Evidence from a study in the tsetse free pastoral zone of Galim.

The role of tabanids as potential transmitters of animal trypanosomiasis (AAT) has not yet been established in Cameroon. The objectives of this study were: (i) to trap and determine the species richness and abundance of tabanids, (ii) to identify circulating trypansomes in cattle and tabanids in a tsetse free area. A three year (2015 to 2017) tabanid survey in six regions of Cameroon was conducted. In Galim village, which is in a tsetse free area, both tabanids and cattle blood samples were screened by PCR for the presence of trypanosome DNA. Tabanids were diverse in Littoral (13 species) and in Adamawa (13 species), but were abundant in the Far North region (36.37 to 145.58 tabanids per trap per day (t/t/d)). In Galim, the tabanid trypanosomal DNA presence was 24.4% (95% CI: 11.25-37.53), while the bovine trypanosomal DNA presence was 4.8% (95% CI: 1.68-11.20). In this village, the Trypanosoma spp. identified in tabanids were T. theileri, T. vivax and T. evansi, while those in cattle were T. theileri and T. vivax. The control of tabanids is required to stop the mechanical spread of AAT in tsetse free areas.

First records of two genera and thirteen species of Tabanidae (Diptera) from Honduras.

This works presents information on the diversity of the Tabanidae of Honduras as a product of the examination of 386 specimens and a literature review. Thirteen species and two genera (Bolbodimyia and Dasychela) are recorded from the country for the first time. Eighty-five species distributed in 22 genera, five tribes, and three subfamilies are now known from Honduras. A key to the subfamilies, tribes, and genera of the known Honduran species is also included. All new records are mapped and illustrated to aid in the identification of the species.

The COMBAT project: controlling and progressively minimizing the burden of vector-borne animal trypanosomosis in Africa.

Vector-borne diseases affecting livestock have serious impacts in Africa. Trypanosomosis is caused by parasites transmitted by tsetse flies and other blood-sucking Diptera. The animal form of the disease is a scourge for African livestock keepers, is already present in Latin America and Asia, and has the potential to spread further. A human form of the disease also exists, known as human African trypanosomosis or sleeping sickness. Controlling and progressively minimizing the burden of animal trypanosomosis (COMBAT) is a four-year research and innovation project funded by the European Commission, whose ultimate goal is to reduce the burden of animal trypanosomosis (AT) in Africa. The project builds on the progressive control pathway (PCP), a risk-based, step-wise approach to disease reduction or elimination. COMBAT will strengthen AT control and prevention by improving basic knowledge of AT, developing innovative control tools, reinforcing surveillance, rationalizing control strategies, building capacity, and raising awareness. Knowledge gaps on disease epidemiology, vector ecology and competence, and biological aspects of trypanotolerant livestock will be addressed. Environmentally friendly vector control technologies and more effective and adapted diagnostic tools will be developed. Surveillance will be enhanced by developing information systems, strengthening reporting, and mapping and modelling disease risk in Africa and beyond. The socio-economic burden of AT will be assessed at a range of geographical scales. Guidelines for the PCP and harmonized national control strategies and roadmaps will be developed. Gender equality and ethics will be pivotal in all project activities. The COMBAT project benefits from the expertise of African and European research institutions, national veterinary authorities, and international organizations. The project consortium comprises 21 participants, including a geographically balanced representation from 13 African countries, and it will engage a larger number of AT-affected countries through regional initiatives.

A nationwide survey of the tabanid fauna of Cameroon.

BACKGROUND: Tabanids are a neglected group of haematophagous dipterans despite containing 4434 species, regrouped in > 144 genera. They are mechanical vectors of important pathogens, including viruses, bacteria and protozoa of humans and domesticated and wild animals. As it is > 50 years since the publication of a preliminary nationwide record of the tabanids of Cameroon identified 84 species, updated information is needed. The aim of this study was to provide current data on the species composition, abundance and distribution of tabanids in the five main agro-ecological zones (AEZs) of Cameroon. METHODS: From 2015 to 2017, a systematic entomological study using Nzi, Vavoua, Biconical and Sevi traps (n = 106) was conducted in 604 trapping points over 11,448 trap-days in the five main AEZs of Cameroon. RESULTS: A total of 25,280 tabanids belonging to 25 species were collected, including eight species not previously documented in Cameroon, namely Tabanus latipes (1 female), Tabanus ricardae (1 female), Tabanus fasciatus (32 females and 6 males), Haematopota pluvialis (18 females), Haematopota decora (19 females and 3 males), Haematopota nigripennis (18 females), Chrysops distinctipennis (47 females and 5 males) and Ancala fasciata (34 females and 7 males). The distribution maps of the newly identified tabanids differed between AEZs, with most tabanids collected from the Guinean savanna. The highest apparent density of tabanids was recorded in the Sudan Savanna region, and the mean apparent densities of species with sites was statistically significantly different (Student t-test: 2.519, df = 24, P = 0.019). The highest species diversity was found in the rainforest. CONCLUSIONS: This study increased the list of tabanids recorded in Cameroon from 84 species in the preliminary record to 92 species, with most of the newly identified species occurring in the Guinea Savanna AEZ. The high diversity and abundance of tabanids in the livestock/wildlife interface areas of the rain forests and Sudan Savanna AEZs, respectively, suggest risk of mechanical transmission of pathogens. Investigations of the microbiota of tabanids in the different AEZs to define their role as disease vectors are proposed.

A Review of Arguments for the Existence of Latent Infections of Bacillus anthracis, and Research Needed to Understand their Role in the Outbreaks of Anthrax.

Hugh-Jones and Blackburn and Turnbull's collective World Health Organization (WHO) report did literature reviews of the theories and the bases for causes of anthrax outbreaks. Both comment on an often-mentioned suspicion that, even though unproven, latent infections are likely involved. Hugh-Jones suggested Gainer do an updated review of our present-day knowledge of latent infections, which was the basis for Gainer's talk at the Biology of Anthrax Conference in Bari, Italy 2019. At the Conference Gainer met Vergnaud who presented anthrax genome studies that implied that the disease might have spread throughout Asia and from Europe to North America in a short time span of three or four centuries. Vergnaud wondered if latent infections might have played a role in the process. Several other presenters at the Conference also mentioned results that might suggest the existence of latent infections. Vergnaud subsequently looked into some of the old French literature about related observations, results, and discussions of early Pasteur vaccine usage (late 1800's) and found mentions of suspected latent infections. The first part of the paper is a focused summary and interpretation of Hugh-Jones and Blackburn's and Turnbull's reviews specifically looking for suggestions of latent infections, a few additional studies with slightly different approaches, and several mentions made of presentations and posters at the Conference in Italy. In general, many different investigators in different areas and aspects of the anthrax study at the Conference found reasons to suspect the existence of latent infections. The authors conclude that the affected species most studied, including Homo sapiens, provide circumstantial evidence of latent infections and modified host resistance. The last part of the review explores the research needed to prove or disprove the existence of latent infections.

Tabanids as possible pathogen vectors in Senegal (West Africa).

BACKGROUND: Species of the Tabanidae are potent vectors of human and animal diseases, but they have not been thoroughly investigated to date. In Senegal (West Africa), little information is available on these dipterans. Our objective in this study was to investigate Senegalese tabanids and their diversity by using molecular and proteomics approaches, as well as their associated pathogens. METHODS: A total of 171 female tabanids were collected, including 143 from Casamance and 28 from Niokolo-Koba. The samples were identified morphologically by PCR sequencing and by MALDI-TOF MS, and PCR analysis was employed for pathogen detection and blood-meal characterization. RESULTS: The morphological identification revealed four species concordantly with the molecular identification: Atylotus fuscipes (79.5%), Tabanus guineensis (16.4%), Chrysops distinctipennis (3.5%) and Tabanus taeniola (0.6%) (not identified by PCR). The molecular investigation of pathogens revealed the presence of Trypanosoma theileri (6.6%), Leishmania donovani (6.6%), Setaria digitata (1.5%), Rickettsia spp. (5.1%) and Anaplasmataceae bacteria (0.7%) in A. fuscipes. Tabanus guineensis was positive for L. donovani (35.7%), S. digitata (3.6%) and Anaplasmataceae (17.8%). Leishmania donovani has been detected in 50% of C. distinctipennis specimens and the only T. taeniola specimen. No Piroplasmida, Mansonella spp. or Coxeilla burnetii DNA was detected. In addition to humans (96.43%), Chlorocebus sabeus, a non-human primate, has been identified as a host of (3.57%) analysed tabanids. MALDI-TOF MS enabled us to correctly identify all tabanid species that had good quality spectra and to create a database for future identification. CONCLUSIONS: Tabanids in Senegal could be vectors of several pathogens threatening animal and public health. To fully characterize these dipterans, it is therefore necessary that researchers in entomology and infectiology employ molecular characterization and mass spectrometric techniques such as MALDI-TOF MS to analyse these dipterans in Senegal and West Africa.

Effect of chitosan on adult longevity when fed, in no-choice experiments, to Musca domestica L., Tabanus nigrovittatus Macquart, and Phormia regina (Meigen) adults and its consumption in adult Musca domestica L.

BACKGROUND: The literature reports that more research needs to be done on using chitosan as an environmentally friendly bioinsecticide, especially against dipterans. Thus, we examined the effect of chitosan when fed, in no-choice experiments, to adult Musca domestica, Tabanus nigrovittatus, and Phormia regina. RESULTS: A 2% chitosan solution was fed, in no-choice experiments, to adults of Musca domestica, Tabanus nigrovittatus, and Phormia regina to study the effects on survivorship. In all species, the uptake of chitosan caused a significant decrease in survivorship. Examination of the digestive tract of house flies showed a thick material within the midgut, plus a shriveling of the midgut only in chitosan-treated flies. A survivorship curve of adult house flies fed a 10% sucrose and 2% w/v ascorbic acid-only solution showed that the effect of the chitosan solution was due to the chitosan and not the ascorbic acid. Intake experiments revealed that by day 2, chitosan treated house flies consumed significantly less of the diet compared to controls. CONCLUSION: Chitosan, a biopesticide, when fed in no-choice experiments to three species of adult flies produced severe mortality within 4-6 days of ingestion. A working hypothesis suggests that by day 2 of the chitosan diet there appears to be a malfunction of the digestive tract and possibly the midgut microbiome. © 2020 Society of Chemical Industry.

New records and DNA barcoding of deer flies, Chrysops (Diptera: Tabanidae) in Thailand.

Chrysops spp. or deer flies (Diptera: Tabanidae) are hematophagous flies of medical and veterinary importance and some species are important vectors of Trypanosoma evansi, the causative agent of surra in Thailand. However, data regarding deer fly species and their molecular identification are limited. Accurate species identification will indicate the appropriate control measures. In this study, an entomological survey of deer flies from different sites in Thailand between May 2018 and June 2019 were conducted. In addition, mitochondrial cytochrome oxidase subunit I (COI) barcoding region was used for species identification. A total of 82 females were collected and 6 species were identified. Of these, three species are new records for Thailand: C. designatus, C. fuscomarginalis and C. vanderwulpi bringing the species total found in Thailand to nine. The COI sequences revealed an intraspecific divergence of 0.0%-2.65% and an interspecific divergence of 7.03%-13.47%. Phylogenetic analysis showed that all deer fly species were clearly separated into distinct clusters according to morphologically identified species. These results indicated that COI barcodes were capable in discriminating between deer fly species on the basis of the barcoding gap and phylogenetic analysis. Therefore, DNA barcoding is a valuable tool for species identification of deer flies in Thailand.

Evaluation of the relative roles of the Tabanidae and Glossinidae in the transmission of trypanosomosis in drug resistance hotspots in Mozambique.

BACKGROUND: Tsetse flies (Diptera: Glossinidae) and tabanids (Diptera: Tabanidae) are haematophagous insects of medical and veterinary importance due to their respective role in the biological and mechanical transmission of trypanosomes. Few studies on the distribution and relative abundance of both families have been conducted in Mozambique since the country's independence. Despite Nicoadala, Mozambique, being a multiple trypanocidal drug resistance hotspot no information regarding the distribution, seasonality or infection rates of fly-vectors are available. This is, however, crucial to understanding the epidemiology of trypanosomosis and to refine vector management. METHODS: For 365 days, 55 traps (20 NGU traps, 20 horizontal traps and 15 Epsilon traps) were deployed in three grazing areas of Nicoadala District: Namitangurine (25 traps); Zalala (15 traps); and Botao (15 traps). Flies were collected weekly and preserved in 70% ethanol. Identification using morphological keys was followed by molecular confirmation using cytochrome c oxidase subunit 1 gene. Trap efficiency, species distribution and seasonal abundance were also assessed. To determine trypanosome infection rates, DNA was extracted from the captured flies, and submitted to 18S PCR-RFLP screening for the detection of Trypanosoma. RESULTS: In total, 4379 tabanids (of 10 species) and 24 tsetse flies (of 3 species), were caught. NGU traps were more effective in capturing both the Tabanidae and Glossinidae. Higher abundance and species diversity were observed in Namitangurine followed by Zalala and Botao. Tabanid abundance was approximately double during the rainy season compared to the dry season. Trypanosoma congolense and T. theileri were detected in the flies with overall infection rates of 75% for tsetse flies and 13% for tabanids. Atylotus agrestis had the highest infection rate of the tabanid species. The only pathogenic trypanosome detected was T. congolense. CONCLUSIONS: Despite the low numbers of tsetse flies captured, it can be assumed that they are still the cyclical vectors of trypanosomosis in the area. However, the high numbers of tabanids captured, associated to their demonstrated capacity of transmitting trypanosomes mechanically, suggest an important role in the epidemiology of trypanosomosis in the Nicoadala district. These results on the composition of tsetse and tabanid populations as well as the observed infection rates, should be considered when defining strategies to control the disease.

Shared species of crocodilian trypanosomes carried by tabanid flies in Africa and South America, including the description of a new species from caimans, Trypanosoma kaiowa n. sp.

BACKGROUND: The genus Trypanosoma Gruby, 1843 is constituted by terrestrial and aquatic phylogenetic lineages both harboring understudied trypanosomes from reptiles including an increasing diversity of crocodilian trypanosomes. Trypanosoma clandestinus Teixeira & Camargo, 2016 of the aquatic lineage is transmitted by leeches to caimans. Trypanosoma grayi Novy, 1906 of the terrestrial lineage is transmitted by tsetse flies to crocodiles in Africa, but the vectors of Neotropical caiman trypanosomes nested in this lineage remain unknown. RESULTS: Our phylogenetic analyses uncovered crocodilian trypanosomes in tabanids from South America and Africa, and trypanosomes other than T. grayi in tsetse flies. All trypanosomes found in tabanids clustered in the crocodilian clade (terrestrial lineage) forming six clades: Grayi (African trypanosomes from crocodiles and tsetse flies); Ralphi (trypanosomes from caimans, African and Brazilian tabanids and tsetse flies); Terena (caimans); Cay03 (caimans and Brazilian tabanids); and two new clades, Tab01 (Brazilian tabanid and tsetse flies) and Kaiowa. The clade Kaiowa comprises Trypanosoma kaiowa n. sp. and trypanosomes from African and Brazilian tabanids, caimans, tsetse flies and the African dwarf crocodile. Trypanosoma kaiowa n. sp. heavily colonises tabanid guts and differs remarkably in morphology from other caiman trypanosomes. This species multiplied predominantly as promastigotes on log-phase cultures showing scarce epimastigotes and exhibited very long flagellates in old cultures. Analyses of growth behavior revealed that insect cells allow the intracellular development of Trypanosoma kaiowa n. sp. CONCLUSIONS: Prior to this description of Trypanosoma kaiowa n. sp., no crocodilian trypanosome parasitic in tabanid flies had been cultured, morphologically examined by light, scanning and transmission microscopy, and phylogenetically compared with other crocodilian trypanosomes. Additionally, trypanosomes thought to be restricted to caimans were identified in Brazilian and African tabanids, tsetse flies and the dwarf crocodile. Similar repertoires of trypanosomes found in South American caimans, African crocodiles and tabanids from both continents support the recent diversification of these transcontinental trypanosomes. Our findings are consistent with trypanosome host-switching likely mediated by tabanid flies between caimans and transoceanic migrant crocodiles co-inhabiting South American wetlands at the Miocene.

Horse flies (Diptera: Tabanidae) of three West African countries: A faunistic update, barcoding analysis and trypanosome occurrence.

Horse flies (Diptera: Tabanidae) are of medical and veterinary importance since they transmit a range of pathogens. The horse fly fauna of tropical Africa is still poorly known, and in some geographical areas has not been studied for decades. This study summarizes the results of tabanid collections performed in three West African countries where only sparse data were previously available, the Central African Republic (CAR), Gabon and Liberia. Of 1093 collected specimens, 28 morphospecies and 26 genospecies belonging to six genera were identified, including the first findings of eleven morphospecies in the countries where horse flies were collected: Philoliche (Subpangonia) gravoti Surcouf, 1908 and Tabanus ianthinus Surcouf, 1907 are new records for Liberia; Ancala fasciata f. mixta (Surcouf, 1914), Tabanus fraternus Macquart, 1846, and T. triquetrornatus Carter, 1915 for CAR; Chrysops longicornis Macquart, 1838, Haematopota albihirta Karsch, 1887, H. bowdeni Oldroyd, 1952, and H. brucei Austen, 1908 for Gabon; and Tabanus secedens f. regnaulti Surcouf, 1912 and T. thoracinus Palisot de Beauvois, 1807 for Gabon and Liberia. Species identification of all 28 morphospecies based on morphological features was further supplemented by barcoding of cytochrome oxidase I (COI). Based on the COI sequences of 115 specimens representing 74 haplotypes, a phylogenetic tree was constructed to illustrate the relationships among the tabanid species found and to demonstrate their intra- and interspecific divergences. Our study enriches the current number of barcoded tabanids with another 22 genospecies. Based on the analysis of molecular data we question the taxonomic relevance of the morphological forms Ancala fasciata f. mixta and Tabanus secedens f. regnaulti. A parasitological survey based on nested PCR of 18S rRNA revealed a high (˜25%) prevalence of Trypanosoma theileri in the studied horse flies, accompanied by two species of monoxenous trypanosomatids, Crithidia mellificae and Blastocrithidia sp.