Factors affecting transmission of trypanosomes through tsetse flies

The maintenance of human sleeping sickness and nagana across sub-Saharan Africa depends on cyclical transmission of trypanosomes through tsetse flies. Infection rates in tsetse are normally very low as most parasites ingested with a bloodmeal die in the fly gut. Infections which successfully establish in the fly midgut may subsequently mature into mammalian infective trypanosomes in the salivary glands. However, these processes are not automatic and involve tsetse, symbiont, trypanosome and environmental factors.Previous work showed that the symbiotic bacterium Sodalis glossinidius was involved in susceptibility to trypanosome infection. Streptozotocin (a toxic analogue of the bacterium's main food source) has been recently shown to decrease trypanosome infection rates in the offspring of treated tsetse. In the present work streptozotocin did remove S. glossinidius from the offspring of treated flies but it was not possible to generate a line of tsetse free from 5. glossinidius infection.Other potential factors involved in acquisition of trypanosome infection were then examined. A range of antioxidants or cyclic GMP were shown to prevent trypanosome death in the tsetse midgut. The process was shown to be independent of protein synthesis as D-cysteine (an unphysiological isomer of L-cysteine) also enhanced midgut infection rates. Further experiments showed that cGMP could significantly inhibit trypanosome death when fed up to 96 h post-infection, whereas antioxidants only functioned for 48 h post-infection. Moreover it was found that maturation of established midgut infections could be regulated by environmental stimuli as well as by antioxidants. Cold shock of infected flies as well as addition of L-cysteine but not D-cysteine to the bloodmeal resulted in significant increases in maturation rates, while nitric oxide synthase inhibitors reduced maturation rates.It is concluded that reactive oxygen species play a major role in killing trypanosomes entering the tsetse midgut and that cysteine containing proteins and/or nitric oxide are essential for differentiation of established midgut infections into mammalian infective salivary gland infections

Effects of cyclic nucleotides on midgut infections and maturation of T. b. brucei in G. m. morsitans

Cyclic nucleotide signalling through cyclic adenosine monophosphate (cAMP) is thought to play an important role in the transformation of the long slender (dividing) form to the short-stumpy (arrested) form in the mammalian bloodstream but the role of cyclic nucleotides in the tsetse-based part of the trypanosome life cycle is unknown. In a series of in vivo experiments, it was found that cyclic guanosine monophosphate (cGMP) but not cAMP could induce significantly higher rates of midgut infection in tsetse. Continuous feeding of either cGMP or cAMP to tsetse had no effect on rates of maturation of established midgut infections suggesting that these two parts of the life cycle in tsetse are not linked

Critical linkages between livestock production, livestock trade and potential spread of human African trypanosomiasis in Uganda:Bioeconomic herd modeling and livestock trade analysis

Background: Tsetse-transmitted human African trypanosomiasis (HAT) remains endemic in Uganda. The chronic form caused by Trypanosoma brucei gambiense (gHAT) is found in north-western Uganda, whereas the acute zoonotic form of the disease, caused by T. b. brucei rhodesiense (rHAT), occurs in the eastern region. Cattle is the major reservoir of rHAT in Uganda. These two forms of HAT are likely to converge resulting in a public health disaster. This study examines the intricate and intrinsic links between cattle herd dynamics, livestock trade and potential risk of spread of rHAT northwards. Methods: A bio-economic cattle herd model was developed to simulate herd dynamics at the farm level. Semi-structured interviews (n = 310), focus group discussions (n = 9) and key informant interviews (n = 9) were used to evaluate livestock markets (n = 9) as part of the cattle supply chain analysis. The cattle market data was used for stochastic risk analysis. Results: Cattle trade in eastern and northern Uganda is dominated by sale of draft and adult male cattle as well as exportation of young male cattle. The study found that the need to import draft cattle at the farm level was to cover deficits because of the herd structure, which is mostly geared towards animal traction. The importation and exportation of draft cattle and disposal of old adult male cattle formed the major basis of livestock movement and could result in the spread of rHAT northwards. The risk of rHAT infected cattle being introduced to northern Uganda from the eastern region via cattle trade was found to be high (i.e. probability of 1). Conclusion: Through deterministic and stochastic modelling of cattle herd and cattle trade dynamics, this study identifies critical links between livestock production and trade as well as potential risk of rHAT spread in eastern and northern Uganda. The findings highlight the need for targeted and routine surveillance and control of zoonotic diseases such as rHAT

Amplified fragment length polymorphism (AFLP) analysis of closely related wild and captive tsetse fly (Glossina morsitans morsitans) populations

<p>Abstract</p> <p>Background</p> <p>Tsetse flies (Diptera: Glossinidae) are vectors of trypanosomes that cause sleeping sickness in humans and nagana in livestock across sub-Saharan Africa. Tsetse control strategies rely on a detailed understanding of the epidemiology and ecology of tsetse together with genetic variation within and among populations. High-resolution nuclear genetic markers are useful tools for elucidation of the genetic basis of phenotypic traits. In this study amplified fragment length polymorphism (AFLP) markers were developed to analyze genetic variation in <it>Glossina morsitans morsitans </it>from laboratory and field-collected populations from Zimbabwe.</p> <p>Results</p> <p>A total of seven hundred and fifty one loci from laboratory and field populations of <it>G. m. morsitans </it>from Zimbabwe were genotyped using AFLP with seven primer combinations. Analysis identified 335 polymorphic loci. The two populations could be distinguished by cluster and principal components analysis (PCA) analysis, indicating that AFLP markers can be used to separate genetically similar populations; at the same time differences observed between laboratory and field populations were not very great. Among the techniques investigated, the use of acetone was the most reliable method of preservation of tsetse for subsequent extraction of high molecular weight DNA. An interesting finding was that AFLP also enabled robust within-population discrimination of male and female tsetse flies due to their different X chromosome DNA complements.</p> <p>Conclusions</p> <p>AFLP represents a useful additional tool to add to the suite of techniques currently available for the genetic analysis of tsetse populations and represents a useful resource for identification of the genetic basis of important phenotypic traits.</p

A comparative evaluation of PCR- based methods for species- specific determination of African animal trypanosomes in Ugandan cattle

BACKGROUND: In recent years, PCR has been become widely applied for the detection of trypanosomes overcoming many of the constraints of parasitological and serological techniques, being highly sensitive and specific for trypanosome detection. Individual species-specific multi-copy trypanosome DNA sequences can be targeted to identify parasites. Highly conserved ribosomal RNA (rRNA) genes are also useful for comparisons between closely related species. The internal transcribed spacer regions (ITS) in particular are relatively small, show variability among related species and are flanked by highly conserved segments to which PCR primers can be designed. Individual variations in inter-species length makes the ITS region a useful marker for identification of multiple trypanosome species within a sample. METHODS: Six hundred blood samples from cattle collected in Uganda on FTA cards were screened using individual species-specific primers for Trypanosoma congolense, Trypanosoma brucei and Trypanosoma vivax and compared to a modified (using eluate extracted using chelex) ITS-PCR reaction. RESULTS: The comparative analysis showed that the species-specific primer sets showed poor agreement with the ITS primer set. Using species-specific PCR for Trypanozoon, a prevalence of 10.5% was observed as compared to 0.2% using ITS PCR (Kappa = 0.03). For Trypanosoma congolense, the species-specific PCR reaction indicated a prevalence of 0% compared to 2.2% using ITS PCR (Kappa = 0). For T. vivax, species-specific PCR detected prevalence of 5.7% compared to 2.8% for ITS PCR (Kappa = 0.29). CONCLUSIONS: When selecting PCR based tools to apply to epidemiological surveys for generation of prevalence data for animal trypanosomiasis, it is recommended that species-specific primers are used, being the most sensitive diagnostic tool for screening samples to identify members of Trypanozoon (T. b. brucei s.l). While ITS primers are useful for studying the prevalence of trypanosomes causing nagana (in this study the species-specific primers did not detect the presence of T. congolense) there were discrepancies between both the species-specific primers and ITS for the detection of T. vivax

Exploring the Effect of Human and Animal Population Growth on Vector-Borne Disease Transmission with an Agent-Based Model of Rhodesian Human African Trypanosomiasis in Eastern Province, Zambia

This paper presents the development of an agent-based model (ABM) to investigate Trypanosoma brucei rhodesiense human African trypanosomiasis (rHAT) disease transmission. The ABM model, fitted at a fine spatial scale, was used to explore the impact of a growing host population on the spread of disease along a 75 km transect in the Luangwa Valley, Zambia. The model was used to gain a greater understanding of how increases in human and domestic animal population could impact the contact network between vector and host, the subsequent transmission patterns, and disease incidence outcomes in the region. Modelled incidence rates showed increases in rHAT transmission in both humans and cattle. The primary demographic attribution of infection switched dramatically from young children of both sexes attending school, to adult women performing activities with shorter but more frequent trips, such as water and firewood collection, with men more protected due to the presence of cattle in their routines. The interpretation of model output provides a plausible insight into both population development and disease transmission in the near future in the region and such techniques could aid well-targeted mitigation strategies in the future

Exploring the Effect of Human and Animal Population Growth on Vector-Borne Disease Transmission with an Agent-Based Model of Rhodesian Human African Trypanosomiasis in Eastern Province, Zambia

This paper presents the development of an agent-based model (ABM) to investigate Trypanosoma brucei rhodesiense human African trypanosomiasis (rHAT) disease transmission. The ABM model, fitted at a fine spatial scale, was used to explore the impact of a growing host population on the spread of disease along a 75 km transect in the Luangwa Valley, Zambia. The model was used to gain a greater understanding of how increases in human and domestic animal population could impact the contact network between vector and host, the subsequent transmission patterns, and disease incidence outcomes in the region. Modelled incidence rates showed increases in rHAT transmission in both humans and cattle. The primary demographic attribution of infection switched dramatically from young children of both sexes attending school, to adult women performing activities with shorter but more frequent trips, such as water and firewood collection, with men more protected due to the presence of cattle in their routines. The interpretation of model output provides a plausible insight into both population development and disease transmission in the near future in the region and such techniques could aid well-targeted mitigation strategies in the future

Sodalis glossinidius prevalence and trypanosome presence in tsetse from Luambe National Park, Zambia

Background Tsetse flies are the biological vectors of African trypanosomes, the causative agents of sleeping sickness in humans and nagana in animals. The tsetse endosymbiont Sodalis glossinidius has been suggested to play a role in tsetse susceptibility to infection. Here we investigate the prevalence of African trypanosomes within tsetse from the Luambe National Park, Zambia and if there is an association between S. glossinidius and presence of trypanosomes within the tsetse examined. Methods Tsetse representing three species (Glossina brevipalpis, Glossina morsitans morsitans and Glossina pallidipes), were sampled from Luambe National Park, Zambia. Following DNA extraction, PCR was used to examine the tsetse for presence of trypanosomes and the secondary endosymbiont S. glossinidius. Results S. glossinidius infection rates varied significantly between tsetse species, with G. brevipalpis (93.7%) showing the highest levels of infection followed by G. m. morsitans (17.5%) and G. pallidipes (1.4%). ITS-PCR detected a wide variety of trypanosomes within the tsetse that were analysed. Significant differences were found in terms of trypanosome presence between the three tsetse species. A high proportion of G. m. morsitans were shown to carry T. brucei s.l. DNA (73.7%) and of these around 50% were positive for Trypanosoma brucei rhodesiense. T. vivax, T. godfreyi, T. simiae, T. simiae Tsavo and T. congolense were also detected. No association was found between the occurrence of S. glossinidius and the presence of trypanosome DNA in any of the three tsetse species tested. Conclusion The current work shows that T. b. rhodesiense was circulating in Luambe National Park, representing a risk for people living in the park or surrounding area and for tourists visiting the park. The differences in trypanosome DNA presence observed between the different tsetse species tested may indicate host feeding preferences, as the PCR will not discriminate between a fly with an active/resident infection compared to a refractory fly that has fed on an infected animal. This makes it difficult to establish if S. glossinidius may play a role in the susceptibility of tsetse flies to trypanosome infection. Keywords: Glossina; Rhodesiense; Wildlife; Morsitans; Pallidipes; BrevipalpisRoyal Zoological Society of Scotland. United Kingdom Department for International Development Animal Health Programme (DFID-AHP)

Livestock network analysis for rhodesiense human African trypanosomiasis control in Uganda

Background: Infected cattle sourced from districts with established foci for Trypanosoma brucei rhodesiense human African trypanosomiasis (rHAT) migrating to previously unaffected districts, have resulted in a significant expansion of the disease in Uganda. This study explores livestock movement data to describe cattle trade network topology and assess the effects of disease control interventions on the transmission of rHAT infectiousness.Methods: Network analysis was used to generate a cattle trade network with livestock data which was collected from cattle traders (n = 197) and validated using random graph methods. Additionally, the cattle trade network was combined with a susceptible, infected, recovered (SIR) compartmental model to simulate spread of rHAT (Ro 1.287), hence regarded as “slow” pathogen, and evaluate the effects of disease interventions.Results: The cattle trade network exhibited a low clustering coefficient (0.5) with most cattle markets being weakly connected and a few being highly connected. Also, analysis of the cattle movement data revealed a core group comprising of cattle markets from both eastern (rHAT endemic) and northwest regions (rHAT unaffected area). Presence of a core group may result in rHAT spread to unaffected districts and occurrence of super spreader cattle market or markets in case of an outbreak. The key cattle markets that may be targeted for routine rHAT surveillance and control included Namutumba, Soroti, and Molo, all of which were in southeast Uganda. Using effective trypanosomiasis such as integrated cattle injection with trypanocides and spraying can sufficiently slow the spread of rHAT in the network.Conclusion: Cattle trade network analysis indicated a pathway along which T. b. rhodesiense could spread northward from eastern Uganda. Targeted T. b. rhodesiense surveillance and control in eastern Uganda, through enhanced public–private partnerships, would serve to limit its spread