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Reliable diagnostic tools are needed to choose the appropriate treatment and proper control measures for animal trypanosomoses, some of which are pathogenic.Trypanosoma cruzi, for example, is responsible for Chagas disease in Latin America. Similarly, pathogenic animal trypanosomoses of African origin (ATAO), including a variety ofTrypanosomaspecies and subspecies, are currently found in Africa, Latin America and Asia. ATAO limit global livestock productivity and impact food security and the welfare of domestic animals. This review focusses on implementing previously reviewed diagnostic methods, in a complex epizootiological scenario, by critically assessing diagnostic results at the individual or herd level. In most cases, a single diagnostic method applied at a given time does not unequivocally identify the various parasitological and disease statuses of a host. These include “non-infected”, “asymptomatic carrier”, “sick infected”, “cured/not cured” and/or “multi-infected”. The diversity of hosts affected by these animal trypanosomoses and their vectors (or other routes of transmission) is such that integrative, diachronic approaches are needed that combine: (i) parasite detection, (ii) DNA, RNA or antigen detection and (iii) antibody detection, along with epizootiological information. The specificity of antibody detection tests is restricted to the genus or subgenus due to cross-reactivity with otherTrypanosomaspp. and Trypanosomatidae, but sensitivity is high. The DNA-based methods implemented over the last three decades have yielded higher specificity and sensitivity for active infection detection in hosts and vectors. However, no single diagnostic method can detect all active infections and/or trypanosome species or subspecies. The proposed integrative approach will improve the prevention, surveillance and monitoring of animal trypanosomoses with the available diagnostic tools. However, further developments are required to address specific gaps in diagnostic methods and the sustainable control or elimination of these diseases.Graphical Abstract

Background and Aim: Trypanosoma evansi infection has been reported in Thai livestock such as beef and dairy cattle. However, there is little information on T. evansi infection in bullfighting cattle in Southern Thailand. The aim of this study was to investigate the infection of T. evansi in bullfighting cattle presented for health checks at the Animal Hospital, Faculty of Veterinary Science, Prince of Songkla University, Thailand. Materials and Methods: Blood and serum samples were collected from 177 bullfighting cattle from April 2016 to February 2017 after bullfighting matches. Animal inspected showed signs of fever, weight loss, or exercise intolerance. Investigation of T. evansi infection was tested using polymerase chain reaction (PCR) with TBR primers and using indirect enzyme-linked immunosorbent assay with T. evansi crude antigen. Results: The seroprevalence of T. evansi in bullfighting cattle was 22.60% (40/177). The PCR results detected no parasite DNA in this study. However, bullfighting cattle may serve as T. evansi reservoirs. Conclusion: Health checking procedures for T. evansi should be promoted for bullfighting events so that infected animals can be quarantined in the preparatory stages of such events.

With an increasing worldwide population that presently exceeds 38 million, camels are important source of meat, milk, and transportation of goods, in many regions of the world. Camels are particularly critical in the northern parts of Africa, above the tsetse belt. However, camel breeding areas are expanding into southern areas, under the pressures of global warming, leading to increasing risk of acquiring parasitic infections in these non-traditional ecotypes. Common biting flies (tabanids, stomoxyine flies, and Hippobosca camelina) act as mechanical vectors, resulting in exposure to trypanosomosis (Trypanosoma evansi; Surra) and high camel morbidity and mortality. In these regions, complicating infections with other Trypanosoma may also occur, particularly Trypanosoma vivax. In many modern camel-breeding areas, human populations are living under political upheaval (terrorism, riots), poverty, and precarity (drought, climate modification). Hence, control and/or elimination of Surra in camels would be beneficial to the economies of these populations. Due to the relatively straightforward epidemiology (single parasite with seasonal transmission in a single host species), control of Surra in Africa is affordable and should be based on implementing: (1) national veterinary services capabilities; (2) efficient diagnosis and control methods; (3) joint integrated control of Surra, gastrointestinal helminthoses (mainly haemonchosis), and sarcoptic mange. We propose that methods to control two economically-critical disease problems, gastrointestinal parasitosis and sarcoptic mange, will support improved Surra control in camels. Aided by decision-makers and donors, elimination of Surra could improve camel health and productivity, and stabilize camel-rearing in regions of the world that suffer from political instability and global warming pressures.

This assessment was conducted by the ad hoc working group ‘ASF vectors’ of the French Agency for Food, Environmental and Occupational Health & Safety (Anses).; International audience; African swine fever (ASF) represents a global threat with huge economic consequences for the swine industry. Even though direct contact is likely to be the main transmission route from infected to susceptible hosts, recent epidemiological investigations have raised questions regarding the role of haematophagous arthropods, in particular the stable fly (Stomoxys calcitrans). In this study, we developed a mechanistic vector-borne transmission model for ASF virus (ASFV) within an outdoor domestic pig farm in order to assess the relative contribution of stable flies to the spread of the virus. The model was fitted to the ecology of the vector, its blood-feeding behaviour and pig-to-pig transmission dynamic. Model outputs suggested that in a context of low abundance (

Animal trypanosomoses due to trypanosomes of African origin (ATAO), mainly caused by Trypanosoma congolense type Savannah (TCS), T. brucei brucei (TBB), T. vivax (TV), and T. evansi, are widespread diseases that affect domestic and wild mammals and have a huge economic impact. ATAO clinical suspicions are usually confirmed by parasitological and molecular methods, while sero-epidemiological surveys are generally carried out using the OIE-recommended ELISA method based on whole cell lysate soluble antigens (WCLSA) from purified trypanosomes; this reagent is usually stored frozen. With a view to expanding this ELISA test, we assessed, standardized, and validated the use of dehydrated rather than frozen WCLSA and serum samples. For the three ELISA assays (TV, TCS & TBB), a repeatability study revealed no significant difference between repeats. The results obtained using frozen rather than freeze-dried antigen and serum strongly correlated for Pearson’s correlation values (>0.93) and Lin’s measure (“very good” to “excellent”). Reproducibility was robust, with Pearson’s correlation values >0.97 for inter technician effects, and 0.87 (TV) to 0.97 (TBB & TCS) for inter-laboratory tests; their combination was “very satisfactory” to “excellent” according to Lin’s measure and there was no impact on qualitative test results. Dehydrated reagents offer the advantage of shipment at room temperature, allowing the secured provision of reagents to regional laboratories. Together with a compendium of standard diagnostic protocols for ATAO (/OIE), dehydrated reagents will enable the serological diagnosis of ATAO at regional level in endemic countries. This very welcome improvement in the context of the Progressive Control Pathway for trypanosomes, recently launched by African countries, will possibly be extended to Latin America in the near future.

Trypanosoma evansi, known as an animal trypanosome, is widely distributed in many countries of Africa, Asia and South America; it causes significant economic loss in these countries. A few cases have also occurred in some countries of Europe due to the importation of infected animals from endemic regions. Rare human T. evansi infections were attended by the health departments and international health organizations in these endemic countries. Trypanosoma lewisi, a cosmopolitan parasite of rats, sometimes found in humans, is currently considered as a zoonotic pathogen and has gained special attention from scientists and international health organizations such as the World Health Organization. The current study considered the serological screening of human infection by T. evansi and T. lewisi in a Chinese human population. None of the 622 samples was found positive for T. evansi infection using the card agglutination test for the trypanosome antigen Rotat 1.2, while, 2.41% of the examined serum samples exhibited some seropositivity to T. lewisi using enzyme-linked immunosorbent assay. No significant difference was found between the samples from areas in the South (Zhaoqing, Guangdong) and Central (Zhengzhou, Henan) China. Keywords: CATT/T. evansi, ELISA T. lewisi, Human infection, Trypanosoma lewisi, T. evansi

We show that the bovid-associated parasite Trypanosoma evansi is endemic in Vietnam and has zoonotic potential. Our study describes the first laboratory-confirmed human case of T. evansi in a previously healthy individual without apolipoprotein L1 deficiency., Background. Trypanosoma is a genus of unicellular parasitic flagellate protozoa. Trypanosoma brucei species and Trypanosoma cruzi are the major agents of human trypanosomiasis; other Trypanosoma species can cause human disease, but are rare. In March 2015, a 38-year-old woman presented to a healthcare facility in southern Vietnam with fever, headache, and arthralgia. Microscopic examination of blood revealed infection with Trypanosoma. Methods. Microscopic observation, polymerase chain reaction (PCR) amplification of blood samples, and serological testing were performed to identify the infecting species. The patient's blood was screened for the trypanocidal protein apolipoprotein L1 (APOL1), and a field investigation was performed to identify the zoonotic source. Results. PCR amplification and serological testing identified the infecting species as Trypanosoma evansi. Despite relapsing 6 weeks after completing amphotericin B therapy, the patient made a complete recovery after 5 weeks of suramin. The patient was found to have 2 wild-type APOL1 alleles and a normal serum APOL1 concentration. After responsive animal sampling in the presumed location of exposure, cattle and/or buffalo were determined to be the most likely source of the infection, with 14 of 30 (47%) animal blood samples testing PCR positive for T. evansi. Conclusions. We report the first laboratory-confirmed case of T. evansi in a previously healthy individual without APOL1 deficiency, potentially contracted via a wound while butchering raw beef, and successfully treated with suramin. A linked epidemiological investigation revealed widespread and previously unidentified burden of T. evansi in local cattle, highlighting the need for surveillance of this infection in animals and the possibility of further human cases.

The reduction in biodiversity from land use change due to urbanization and agricultural intensification appears to be linked to major epidemiological changes in many human diseases. Increasing disease risks and the emergence of novel pathogens result from increased contact among wildlife, domesticated animals, and humans. We investigated the relationship between human alteration of the environment and the occurrence of generalist and synanthropic rodent species in relation to the diversity and prevalence of rodent-borne pathogens in Southeast Asia, a hotspot of threatened and endangered species, and a foci of emerging infectious diseases. We used data from an extensive pathogen survey of rodents from seven sites in mainland Southeast Asia in conjunction with past and present land cover analyses. At low spatial resolutions, we found that rodent-borne pathogen richness is negatively associated with increasing urbanization, characterized by increased habitat fragmentation, agriculture cover and deforestation. However, at a finer spatial resolution, we found that some major pathogens are favored by environmental characteristics associated with human alteration including irriga- tion, habitat fragmentation, and increased agricultural land cover. In addition, synanthropic rodents, many of which are important pathogen reservoirs, were associated with fragmented and human-dominated landscapes, which may ultimately enhance the opportunities for zoonotic transmission and human infection by some pathogens.

Background The main hematophagous flies acting as mechanical vectors of pathogens in mammals are tabanids, stomoxyine flies and some hematophagous Musca spp. Species-identification using dichotomous keys is based on external morphology of the flies. Their proper use requires specific skills and confirmation by shipment of specimens to specific taxonomic area specialists. Although potentially very high (especially in tabanids: 4400 species), the species-diversity of biting flies in livestock is generally limited in a given area (on average 5–10 species). A quick and user-friendly identification would be useful to veterinary technicians to fill gaps in their knowledge and support any control program. Method Based on the facilities brought by numeric photography and the most frequent criteria used in the dichotomous entomologic keys, we generated guidelines to easily create a complete illustration of hematophagous flies external morphology, including dorsal, ventral and lateral views of whole body, head, antenna, thorax, wings, abdomen, legs and some other details, when necessary. Results Application was made through a series of 32 pictures allowing user-friendly description and identification of the female Tabanus rubidus, a common mechanical vector of pathogens in Asia. Conclusion These guidelines will help generating illustrated descriptions of hematophagous flies, which will ease their identification, even at distance. Further on, genotyping of the insects might be a nice complement. Once adopted, these guidelines might be a basis for a better knowledge of the identity, biology and ecology of the hematophagous flies encountered in livestock, and thus, for the development of adapted control measures.

SUMMARYTrypanosoma evansi, the causative agent of surra, is widespread in domestic livestock and wildlife in South East Asia. Surra can affect cattle, buffaloes, horses and also Asian elephants (Elephas maximus). Despite the ‘threatened to extinction’ CITES status of elephant, surra's impact has not been thoroughly assessed yet in this species. This work offers to adapt an antibody enzyme-linked immunosorbent assay (ELISA) protocol, to detect Trypanosoma evansi antibodies in elephant serum. The test was validated with 365 negative-reference samples, which allowed the determination of a 16% positive threshold. The test was applied to a serological survey including 375 individuals. The estimated global seroprevalence was 2·1% (95% CI 1·1–4·2%). Therefore, surra does not appear to be endemic in Thai domestic elephants, but occasional outbreaks were reported to our laboratory during the survey period. These outbreaks seemed to be linked to close proximity to cattle or buffaloes, and led to severe clinical signs in elephants. Frequent relapses were observed after treatment with diminazene aceturate, the only trypanocide drug currently available in Thailand. Therefore, care should be taken to keep elephants away from bovine reservoirs, and to monitor the disease in this endangered species. ELISA proved to be reliable for screening purposes as well as for post-treatment monitoring.

Trypanosoma (Herpetosoma) lewisi is a cosmopolitan parasite of rodents strongly linked to the human dispersal of Rattus spp. from Asia to the rest of the world. This species is highly phylogenetically related to trypanosomes from other rodents (T. lewisi-like), and sporadically infects other mammals. T. lewisi may opportunistically infect humans, and has been considered an emergent rat-borne zoonosis associated to poverty. We developed the THeCATL-PCR based on Cathepsin L (CATL) sequences to specifically detect T. (Herpetosoma) spp., and assess their genetic diversity. This method exhibited high sensitivity using blood samples, and is the first molecular method employed to search for T. lewisi in its flea vectors. THeCATL-PCR surveys using simple DNA preparation from blood preserved in ethanol or filter paper detected T. lewisi in Rattus spp. from human dwellings in South America (Brazil and Venezuela), East Africa (Mozambique), and Southeast Asia (Thailand, Cambodia and Lao PDR). In addition, native rodents captured in anthropogenic and nearby human settlements in natural habitats harbored T. (Herpetosoma) spp. PCR-amplified CATL gene fragments (253 bp) distinguish T. lewisi and T. lewisi-like from other trypanosomes, and allow for assessment of genetic diversity and relationships among T. (Herpetosoma) spp. Our molecular surveys corroborated worldwide high prevalence of T. lewisi, incriminating Mastomys natalensis as an important carrier of this species in Africa, and supported its spillover from invader Rattus spp. to native rodents in Brazil and Mozambique. THeCATL-PCR provided new insights on the accurate diagnosis and genetic repertoire of T. (Herpetosoma) spp. in rodent and non-rodent hosts, revealing a novel species of this subgenus in an African gerbil. Phylogenetic analysis based on CATL sequences from T. (Herpetosoma) spp. and other trypanosomes (amplified using pan-trypanosome primers) uncovered rodents harboring, beyond mammal trypanosomes of different subgenera, some species that clustered in the lizard-snake clade of trypanosomes.

SUMMARYThis study investigated the molecular prevalence of Trypanosoma lewisi and T. evansi in wild rodents from Cambodia, Lao PDR and Thailand. Between 2008 and 2012, rodents (and shrews) were trapped in nine locations and 616 of these were tested using three sets of primers: TRYP1 (amplifying ITS1 of ribosomal DNA of all trypanosomes), TBR (amplifying satellite genomic DNA of Trypanozoon parasites) and LEW1 (amplifying ITS1 of ribosomal DNA of T. lewisi). Based on the size of the PCR products using TRYP1, 17% were positive for T. lewisi and 1·0% positive for Trypanozoon. Results were confirmed by sequencing PCR products and by using more specific primers (LEW1 and TBR). The specificity of TRYP1 primers, however, failed as rodent DNA was amplified in some instances, giving unexpected product sizes. Using LEW1 primers, 13·3% of the samples were confirmed positive for T. lewisi, both by PCR and sequencing. In Thailand, T. lewisi was found in Rattus tanezumi, R. exulans and Berylmys; in Lao PDR, in R. tanezumi and R. exulans, and in Cambodia in R. tanezumi, R. exulans and R. norvegicus. Using TBR, 1·3% of the samples tested positive for Trypanozoon by PCR and sequencing; T. evansi is the only species of the Trypanozoon subgenus possibly present in wild Asian rodents. These results confirmed its presence in rodents from Thailand (R. tanezumi), Lao PDR (R. tanezumi, R. nitidus) and Cambodia (R. tanezumi, Niviventer fulvescens, Maxomys surifer). Based on the information related to rodent trapping, it was found that rodent species trapped in and around human dwellings had a higher prevalence of T. lewisi infection. R. tanezumi and R. exulans, two synanthropic species, were mainly found infected in this habitat suggesting a role as a reservoir and thus a potential source of T. lewisi for human infection.

Trypanosoma lewisi has widely been considered as a non-pathogenic rat trypanosome. However, more and more cases of humans infected with T. lewisi have been reported around the world, indicating that it can infect humans in some undetermined circumstances. Quick and sensitive diagnosis of infection by T. lewisi is important for both treatment of patients and epidemiological studies of this parasite. In this paper, three methods i.e. wet blood smear (diagnosis by microscopy), PCR and LAMP were used to detect T. lewisi from 238 wild rats (Rattus norvegicus) collected from the field in Huadu, Guangdong province, China. Infection rates of these samples detected by the 3 methods was 6.7% (16/238), 12.6% (30/238), and 18.9% (45/238), respectively. LAMP could detect all samples shown positive by microscopical observation of wet smear and by single PCR indicating good potential for application in the detection of T. lewisi. So far as we know, this is the first report of the LAMP method being used to detect T. lewisi in wild rats. The specific T. lewisi LAMP primers were able to amplify the target fragment from the genomic DNA of 19 T. lewisi strains isolated from Huadu, Guangdong province (n = 16), Changchun, Jilin province of China (n = 1) and from Thailand (n = 2). Based on the analyses of ITS1 (internal transcribed spacer 1) and ITS2 sequences, these 19 strains show a very close genetic relationship with over 96–97% similarity to the other corresponding sequences of T. lewisi published in Genbank. Phylogenetic trees of the species in the subgenus Herpetosoma were constructed, based on the ITS1 and ITS2 sequences, and these results also indicate that they are closely related and in the same clade.

Based on the sequence of mitogen-activated protein kinase 5 (TbMAPK5) gene, we have developed a specific PCR method which can delineate the species within the Trypanozoon subgenus from other parasites or host DNA conveniently. In view of further application in field studies, we performed loop-mediated isothermal amplification (LAMP) employing filter paper flecked with rodent blood. Our data showed that TbMAPK5-specific LAMP was sensitive enough to detect a very low parasitemia during the early stages of the infection and fluctuating parasitemia period, and is below the detection limit by microscope. The detection limit for the infected blood sample was 1000 trypanosomes/ml of blood. Based on these results, we consider that TbMAPK5 locus may be a useful target for LAMP diagnosis providing sensitivity and the potential for the genotyping/identification of Trypanozoon species.

Trypanosoma lewisi is a mild or non-pathogenic parasite of the sub-genus Herpetosoma transmitted by fleas to rats. In a previous study we described pan-trypanosome specific primers TRYP1 which amplify the ITS1 of ribosomal DNA by hybridizing in highly conserved regions of 18S and 5.8S genes. These primers proved to be useful for detecting T. lewisi DNA in laboratory rats, but a recent large scale survey in wild rodents demonstrated a lack of specificity. In the present study, we designed and evaluated mono-specific primers LEW1S and LEW1R, for the detection and identification of T. lewisi by a single-step PCR. These primers were designed inside the highly variable region of the ITS1 sequence of T. lewisi ribosomal DNA. The product size of 220 bp is specific to T. lewisi. The sensitivity limit was estimated between 0.055 and 0.55 pg of DNA per reaction, equivalent to 1-10 organisms per reaction. All the PCR products obtained from 6 different T. lewisi isolates were more than 98% similar with each other and similar to the sequences of T. lewisi already published in Genbank. All DNA of 7 T. lewisi stocks from China gave the specific 220 bp product. We showed that LEW1S and LEW1R primers enabled sensitive detection and identification of T. lewisi infection in laboratory and wild rats. This assay is recommended for monitoring T. lewisi infections in rat colonies or for studying infections in the wild fauna. An absence of cross reaction with human DNA means that these primers can be used to investigate atypical trypanosome infections in humans. Given the risk of T. lewisi infection in human, we believe that these primers will be beneficial for public health diagnosis and rodents investigation programmes.

SUMMARYMelarsomine hydrochloride can cure Trypanosoma evansi infection in camels at a dose of 0·25 mg/kg, but at that dose relapses occur in cattle. In our study, the efficacy of an intramuscular injection of melarsomine hydrochloride at 0·5 mg/kg was assessed in 3 normal and 3 splenectomized dairy cattle experimentally infected with a stock of T. evansi from Thailand. The animals were monitored for 5 months by haematocrit centrifugation, blood- or cerebrospinal fluid-mouse inoculation, polymerase chain reaction, the card agglutination test (CATT) for T. evansi, and the enzyme-linked immunosorbent assay‑T. evansi. Parasitological and DNA tests became and remained negative just after treatment. By the end of the experiment, CATT was negative and ELISA scores were below or very close to the cut-off value. One of the splenectomized cattle died from anaplasmosis during the experiment, but tested negative for surra. It was concluded that the parasites had been cleared from the cattle, and melarsomine hydrochloride at 0·5 mg/kg can be recommended for treatment against T. evansi infection in dairy cattle in Thailand. Further work is necessary to validate the efficacy of the treatment in the event of confirmed CSF-infection.

Beside typical human trypanosomes responsible of sleeping sickness in Africa and Chagas disease in Latin America, there is a growing number of reported atypical human infections due to Trypanosoma evansi, a livestock parasite, or Trypanosoma lewisi, a rat parasite, especially in Asia. Drugs available for the treatment of T. brucei ssp. in humans are obviously of choice for the control of T. evansi because it is derived from T. brucei. However, concerning T. lewisi, there is an urgent need to determine the efficacy of trypanocidal drugs for the treatment in humans. In a recent study, pentamidine and fexinidazole were shown to have the best efficacy against one stock of T. lewisi in rats. In the present study suramin, pentamidine, eflornitine, nifurtimox, benznidazole and fexinidazole, were evaluated at low and high doses, in single day administration to normal rats experimentally infected with a stock of T. lewisi recently isolated in Thailand. Because none of these treatments was efficient, a trial was made with the most promising trypanocide identified in a previous study, fexinidazole 100 mg/kg, in 5 daily administrations. Results observed were unclear. To confirm the efficacy of fexinidazole, a mixed infection protocol was set up in cyclophosphamide immunosuppressed rats. Animals were infected successively by T. lewisi and T. evansi, and received 10 daily PO administrations of 200 mg/kg fexinidazole. Drastic effects were observed against T. evansi which was cleared from the rat's blood within 24 to 48 h; however, the treatment did not affect T. lewisi which remained in high number in the blood until the end of the experiment. This mixed infection/treatment protocol clearly demonstrated the efficacy of fexinidazole against T. evansi and its inefficacy against T. lewisi. Since animal trypanocides were also recently shown to be inefficient, other protocols as well as other T. lewisi stocks should be investigated in further studies.

A 20 kg German shepherd dog was presented to a French veterinary teaching hospital for seizures and hyperthermia. The dog had returned 1 month previously from a six-month stay in Senegal and sub-Saharan Africa. Biochemistry and haematology showed severe hypoglycaemia (0.12 g/L), anaemia and thrombocytopenia. Despite administration of large amounts of glucose (30 mL of 30% glucose IV and 10 mL of 70% sucrose by gavage tube hourly), 26 consecutive blood glucose measurements were below 0.25 g/L (except one). Routine cytological examination of blood smears revealed numerous free extracytoplasmic protozoa consistent with Trypanosoma congolense. PCR confirmed a Trypanosoma congolense forest-type infection. Treatment consisted of six injections of pentamidine at 48-hour intervals. Trypanosomes had disappeared from the blood smears four days following the first injection. Clinical improvement was correlated with the normalization of laboratory values. The infection relapsed twice and the dog was treated again; clinical signs and parasites disappeared and the dog was considered cured; however, 6 years after this incident, serological examination by ELISA T. congolense was positive. The status of this dog (infected or non-infected) remains unclear. Hypoglycaemia was the most notable clinical feature in this case. It was spectacular in its severity and in its refractory nature; glucose administration seemed only to feed the trypanosomes, indicating that treatment of hypoglycaemia may in fact have been detrimental.

Antigenic similarities between salivarian trypanosomes are known for a long time, but similarities between salivarian and stercorarian trypanosomes have been very little investigated. Phylogenetically, these genus and species appear to be far. However, in a preliminary work we had shown strong reactions of chagasic human sera using T. evansi antigens in Western-blotting and ELISA. In the current work an ELISA test using T evansi crude antigens was probed with one hundred and two sera of chagasic Bolivian patients previously diagnosed which presented different pathologies. The sensitivity of the ELISA T. evansi was 92.6% similar to that of ELISA T. cruzi. The specificity evaluated using 20 sera of patients infected by Leishmania sp. reaches a comparable value of that obtained with the T. cruzi immunofluorescent assay. Finally, the sensitivity and the specificity of the ELISA T. evansi were not really different from conventional serology of Chagas. In spite of their taxonomic position in various sections and their old divergence, these observations prove a strong antigenic community between T. cruzi and T. evansi. Consequently, the common antigens which remain to be characterized, could be an alternative source of antigen for the detection of antibodies against T. cruzi. Given that T. evansi seems to have strong antigenic communities with the majority of the pathogenic current trypanosomoses of mammals, it is very attractive to identify and characterize these highly conserved antigens which could be suitable targets to develop tools for diagnosis, prophylaxy and chemotherapy against several human and animal trypanosomoses. © 2007 Elsevier B.V. All rights reserved.

A growing number of atypical human infections due to the livestock parasite Trypanosoma evansi, or to the rat parasite Trypanosoma lewisi, are reported in humans in Asia. In some cases, clinical evolutions request treatments, however, so far, there were very few attempts to control T. lewisi using trypanocidal drugs. In a study published elsewhere, the efficacy of human trypanocides is evaluated in laboratory rats, and it concludes that none of them is able to cure rats experimentally infected with T. lewisi. Control of T. lewisi in rat would be a step for identification of drugs against this parasite. In the present study, 4 veterinary drugs: diminazene aceturate, isometamidium chloride, melarsomine hydrochloride and quinapyramine sulfate and chloride, were evaluated at low and high doses, in intra-muscular injections to normal rats experimentally infected with a stock of T. lewisi from Thailand. None of these treatments being efficient, a trial was also made using melarsomine hydrochloride in T. evansi infected rats and in mixed T. lewisi and T. evansi infected rats, in order to demonstrate the efficacy of the drugs under the present protocol. T. evansi was cleared from the rat's blood the day after the treatment, while, T. lewisi remained unaffected until the end of the experiment. These observations clearly demonstrated the efficacy of melarsomine hydrochloride against T. evansi and its inefficacy against T. lewisi. In conclusion none of the veterinary drugs was efficient against this stock of T. lewisi. Other protocols using higher doses or other drugs and T. lewisi stocks should be investigated in further studies. The control of T. lewisi infection in Wistar rats, using veterinary trypanocidal drugs, remains so far unsuccessful.

International audience; Trypanosoma evansi is a protozoan blood parasite and etiologic agent of “surra,” a disease affecting a wide range of domestic and wild mammals, some identified as potential reservoirs. Although T. evansi has been detected in several small wild rodent species, their role in the epidemiology of surra is unclear. There is molecular evidence of T. evansi in wild rodents in Asia, but it is not known whether they can carry the parasite for sufficient time to significantly contribute to the epidemiology of surra. We assessed the susceptibility of the Oriental house rat (OHR; Rattus tanezumi) to T. evansi infection. Five adult male OHRs trapped in Bangkhen district, Bangkok, Thailand, and five laboratory Wistar rats (Rattus norvegicus) as positive controls, were experimentally infected with a local strain of T. evansi. The five controls and three of the five OHRs were highly susceptible and rapidly exhibited the high levels of parasitemia usually observed in Wistar rats. They died or were euthanized just prior to expected death. Two OHRs presented fluctuating levels of parasitemia, without obvious clinical signs, throughout 40 d of monitoring. These results highlight the moderate susceptibility of some OHRs and their ability to carry the infection over time. Along with the molecular evidence of T. evansi in captured OHRs (demonstrated elsewhere), our results bring new information on the potential role of OHRs in the complex epidemiology of surra.

Background Surra, a vector borne disease caused by Trypanosoma (T.) evansi, affects the health, productivity and working capacity of camels. Since clinical signs are not pathognomonic, diagnosis must be confirmed by laboratory methods. This is a first study on the prevalence of surra in Cholistan Desert, Pakistan using a broad variety of diagnostic tests thereby emphasizing it as a risk for the dromedaries of Pakistan. Methods In a cross sectional study, 1005 dromedary camels from three districts in the Cholistan Desert were sampled to assess the prevalence of trypanosomosis due to T. evansi by means of parasitological (Giemsa stained thin smear), serological (formol gel test, CATT/T. evansi, ELISA/VSG RoTat 1.2, immune trypanolysis) and molecular tests (TBR1/2 PCR and RoTat 1.2 PCR). Kappa was calculated to assess the degree of agreement between different tests whereas chi-square test along with odds ratios and their 95 % confidence intervals were used to study influence of breed, gender, age and locality on disease prevalence. Results Overall prevalence was 0.7 % with Giemsa stained thin smears (GST), 40.1 % with formol gel test (FGT), 47.7 % with CATT/T. evansi, 44.2 % with ELISA/VSG RoTat 1.2, 39.9 % with immune trypanolysis (TL), 31.9 % with TBR1/2 PCR and 30.5 % with RoTat1.2 PCR. Based on these results, the Cholistan Desert appears to be a high risk area for surra. According to TL and TBR1/2 PCR, camels at Bahawalpur are approximately two times more likely to be infected than those in Bahawalnagar (OR = 1.8; 95 % CI: 1.38-2.42) and Rahim Yar Khan (OR = 1.9; 95 % CI: 1.30-2.75). Test agreement of TL was moderate with CATT/T. evansi (k = 0.43; 95 % CI: 0.378-0.489) and ELISA/VSG RoTat 1.2 (k = 0.54; 95 % CI: 0.489-0.594) and poor with the other tests. Test agreement between TBR1/2 PCR and RoTat1.2 PCR was almost perfect (k = 0.96; 95 % CI: 0.950-0.984). We didn't find evidence for the presence of T. evansi type B in the studied population. Conclusion Our study supports using antibody detection tests, rather than parasitological and molecular examination, to assess surra prevalence in camels. It also calls for implementation of measures to control surra in the Cholistan Desert.

In order to assess the prevalence of Trypanosoma evansi, a parasitological, molecular, and serological-based investigation wascarried out in randomly sampled horses (n = 375) belonging to different age groups, sexes, and localities from the district of Bahawalpur,Pakistan. The diagnostic performance of applied tests was also compared. The prevalence was recorded as 0.5% with Woo's test, 1.3%with both ITS CF/BR PCR and RoTat 1.2 PCR, and 14.4% with CATT/T.evansi. Based on CATT/T.evansi, significant differences (P ≤0.05) were observed for prevalence estimates according to different localities, sexes, body condition scores, and origins. A nonsignificantdifference (P ≥ 0.05) was observed among different age groups and variable parity numbers. Our study declares district Bahawalpur tobe a high risk area for surra and proposes the potential use of CATT/T.evansi as a field test of choice for surveys in horses. However, thestatus of seropositive animals should be confirmed using a more sensitive molecular tool such as the satellite DNA target. A widespreadstatus of anti-Trypanosoma antibodies calls for control measures and further investigation of various species (camels, cattle, sheep, goats,buffaloes) inhabiting the same area to identify reservoir status.

The trypanosomes pathogenic to livestock in Africa (Trypanosoma congolense, Trypanosoma vivax, and Trypanosoma brucei) are mainly cyclically transmitted by tsetse (Glossina). However, T. vivax, can also be mechanically transmitted by haematophagous insects. Laboratory studies have demonstrated the mechanical transmission of T. congolense, but confirmation of this under natural conditions was necessary. An experiment was therefore carried out in Lahirasso, Burkina Faso, in a corral completely covered by mosquito net, to avoid exposure to tsetse. Eight receiver heifers, free of trypanosome infection, were kept together with two donor heifers, experimentally infected with local stocks of T. congolense. On average, 291 Atylotus agrestis, freshly captured in Nzi traps, were introduced into the mosquito net daily for a period of 20 days to initiate mechanical transmission among cattle. Daily microscopical observation of their blood indicated that two of the eight receiver heifers became infected with T. congolense from days 42 and 53. Mechanical transmission of T. congolense by A. agrestis was demonstrated unequivocally with a 25% incidence over a 20-day period of exposure under a mean challenge of 29 insects/animal/day. These results, in addition to previous reports, demonstrate the ability of A. agrestis to transmit T. vivax and T. congolense to cattle in Africa by mechanical means. Efforts to eliminate cattle trypanosomosis should therefore consider the eventual persistence of disease as a result of mechanical transmission of trypanosomes by tabanids. Index descriptor and abbreviations: Trypanosoma congolense (Trypanosomatidae) is a pathogenic trypanosome found in wild and domestic herbivores, principally in cattle (Bos taurus, Bos indicus, and cross-breds), in Africa. It is cyclically transmitted by tsetse (Glossina, Diptera); however, mechanical transmission by biting insects may also occur. The present study demonstrates unequivocally the mechanical transmission of T. congolense to cattle by one of the most common African tabanids, A. agrestis. The main conclusion is that tabanids are able to transmit T. congolense; however, the incidence of transmission was lower than in studies carried out under the same conditions with T. vivax. Better models of mechanical transmission are required to understand why, on the one hand, epidemiological studies support the mechanical transmission of T. vivax but not T. congolense, and, on the other hand, experimental studies confirm that both species can be mechanically transmitted. Our studies suggest that the epidemiology of trypanosomosis in cattle involves tabanids, and hence, the eradication of tsetse-flies in Africa will not necessarily lead to the eradication of trypanosomosis in domestic livestock. ADT, apparent density of insects per trap per day (mean number of insects caught in one type of trap per 24h of trapping); D, day; NS, not statistically significant

Human-infectious trypanosomes such as Trypanosoma cruzi, T. brucei rhodesiense, and T. b. gambiense can be discriminated from those only infecting animals by their resistance to normal human serum (NHS). These parasites are naturally resistant to trypanolysis induced by the human-specific pore-forming serum protein apolipoprotein L1 (ApoL-1). T. lewisi, a worldwide distributed parasite, has been considered as rat-specific and non-pathogenic to the natural hosts. Here we provide evidence that 19 tested T. lewisi isolates from Thailand and China share resistance to NHS. Further investigation on one selected isolate CPO02 showed that it could resist at least 90% NHS or 30 μg/ml recombinant human ApoL-1 (rhApoL-1) in vitro, in contrast to T. b. brucei which could not survive in 0.0001% NHS and 0.1 μg/ml rhApoL-1. In vivo tests in rats also demonstrated that this parasite is fully resistant to lysis by NHS. Together with recent reports of atypical human infection by T. lewisi, these data allow the conclusion that T. lewisi is potentially an underestimated and thus a neglected human pathogen.

In Senegal, several areas provide great potential for agriculture and animal production, but African animal trypanosomosis (AAT) is one of the major constraints to the development of more effective livestock production systems. A study was conducted to assess the current situation of AAT in this country. Surveys were carried out between June 2011 and September 2012 in four different areas: Dakar, Sine Saloum, Kedougou region and Basse Casamance in several animal species: dogs (152), donkeys (23), horses (63), sheep (43), goats (52) and cattle (104), distributed in the four sites. Molecular tools (PCR) indicated 3.4% positive animals including dogs, donkeys, a goat and cattle. The savannah type of Trypanosoma congolense Broden, 1904 (53% of positive cases) and the forest type of T. congolense (subgenus Nannomonas Hoare, 1964) were predominant. Trypanosoma vivax Ziemann, 1905 (subgenus Duttonella Chalmers, 1918) was only present in one animal and no trypanosome of the subgenus Trypanozoon Lühe, 1906 was found. Half of the positive cases were detected in Sine Saloum, where T. congolense savannah-type was predominant, and the other half in Basse Casamance, where T. congolense forest-type was predominant; no cases were found in Dakar or in the Kedougou region. A high risk of infection in dogs with T. congolense savannah-type was shown in Sine Saloum, requiring prevention and control of dogs in this area. The involvement of tsetse flies in the transmission of T. congolense in Sine Saloum and Basse Casamance is discussed.

International audience; Reports of atypical human cases of Trypanosoma lewisi or T. lewisi-like and Trypanosoma evansi infections have increased in South-East Asia, urging to investigate the possible links between humans, animal reservoirs and habitats. We tested how habitat structure affects the infection by Trypanosoma species of common murine rodents, inhabiting human-dominated landscapes in South East Asia. For this, we used geo-referenced data of rodents investigated for Trypanosoma infection and land cover maps produced for seven study sites in Thailand, Cambodia and Lao PDR. High prevalence of infection by T. lewisi was observed in rodents living near human settlement and in areas with high cover of built-up habitat, while the infection of rodents by T. evansi was explained by increased landscape patchiness and high cover of rain-fed agriculture lands. These results suggest a likely role of wild rodents as reservoir and possible source of atypical human infection by animal trypanosomes.

Une enquete parasitologique et serologique sur les trypanosomoses des bovins a ete realisee dans le secteur nord de la zone de Sideradougou (Burkina Faso), en novembre et decembre 1997. Partant d'un recensement terrestre exhaustif, un millier de bovins ont ete examines sur la base d'un echantillonnage stratifie aleatoire. L'âge et la race des animaux, la nature et la date du dernier traitement trypanocide ont ete enregistres. Les examens parasitologiques ont ete effectues par la methode du "buffy coat", la mesure de l'hematocrite a ete recueillie et les tests Elisa-indirect ont ete realises dans trois systemes differents, avec sensibilisation des plaques par les antigenes solubles de Trypanosoma vivax, T. brucei ou T. congolense (type savane). La technique parasitologique a fourni 5,3 p. cent d'echantillons positifs, domines par l'espece T. congolense. Les examens serologiques ont indique une seroprevalence de 81,7 p. cent (± 2,4 p. cent) les trois tests confondus et, sur la base des resultats chez les animaux âges d'un an, une incidence annuelle moyenne de 52 p. cent (± 11p. cent). L'etude du score maximum de positivite aux trois tests a permis d'estimer les seroprevalences par espece a 79 p. cent pour T. vivax, 3 p. cent pour T. brucei, et 28 p. cent pour T. congolense. L'analyse des donnees serologiques et parasitologiques, selon la date du dernier traitement trypanocide indique par l'eleveur, montre que ces traitements ont peu d'effet sur le taux de portage des anticorps et des parasites, ce dernier est presume assez proche de la seroprevalence observee. La methode du "buffy coat" est insuffisamment sensible pour mesurer la prevalence des infections dans ce type de situation enzootique, mais, associee a la valeur de l'hematocrite, elle a permis d'estimer l'importance des cas de trypanosomose maladie a environ 15 p. cent. Les trypanosomoses bovines demeurent une preoccupation majeure de l'elevage dans la zone agropastorale de Sideradougou. elles sevissent de maniere enzootique avec une predominance nette des infections par T. vivax, une prevalence et un impact clinique eleves de T. congolense. Les donnees generees par cette enquete seront integrees dans un systeme d'information geographique (Sig) mis en place dans la zone pour l'evaluation du risque trypanosomien. (Resume d'auteur)

Les techniques immuno-enzymatiques (enzyme-linked immunosorbent assay : ELISA) utilisées pour le diagnostic des maladies infectieuses ont fait l'objet de nombreuses études de standardisation visant une meilleure reproductibilité des tests, l'expression des résultats, le choix du seuil de positivité et des échantillons de référence. Il apparaît que la standardisation internationale des réactifs et des protocoles est nécessaire pour permettre les contrôles de qualité et la comparaison des résultats entre laboratoires, mais que l'interprétation des résultats peut se heurter à des différences importantes selon le secteur géographique étudié. Partant de ces travaux, et à la lumière du modèle de l'ELISA indirecte pour la recherche des anticorps dirigés contre #Trypanosoma vivax# chez les bovins, l'auteur propose de réaliser la standardisation internationale des réactifs, du protocole et du mode d'expression des résultats ELISA à l'aide d'échantillons de référence internationaux. En revanche, pour la standardisation locale, il est proposé de réaliser : l'échantillonnage des populations de référence locales ; l'établissement des fréquences de distribution des populations locales infectée et non infectée ; le choix de témoins représentatifs des populations locales (échantillons de référence secondaires) ; l'expression des résultats de la réaction par rapport à ces témoins ; l'établissement d'un contrôle interne de qualité fondé sur la réponse des témoins ; la détermination du seuil de positivité selon les exigences de l'utilisateur ; l'adaptation du seuil de positivité selon la prévalence observée dans le secteur géographique étudié. Ces mesures permettent de déterminer la sensibilité et la spécificité du test dans la population étudiée et, lorsque la prévalence de l'infection est connue, de calculer les valeurs prédictives du test.

Les stomoxes sont des vecteurs mécaniques de pathogènes présents dans le sang et les tissus cutanés de leurs hôtes, spécialement le bétail, mais aussi parfois les humains. Pour le bétail, leurs effets directs sont principalement la perturbation des animaux, les lésions de la peau, la réduction de l'alimentation, le stress, la spoliation sanguine et un effet immunosuppressif global. Ils entrainent aussi le regroupement des animaux pour une protection mutuelle ; tout cela favorise le développement des parasites chez les hôtes et leur transmission. Leur effet indirect est la transmission mécanique de pathogènes. En cas de repas interrompu, les stomoxes peuvent reprendre leur repas de sang sur un autre hôte. En injectant de la salive avant l'absorption de sang, ils peuvent inoculer du sang infecté qui restait sur leurs pièces buccales. En plus de cette transmission immédiate, il a été observé que les stomoxes pouvaient conserver du sang dans leur jabot, qui offre un meilleur environnement pour les pathogènes. Ces derniers peuvent être régurgités lors du prochain repas de sang. Ainsi, une transmission retardée semble possible par les stomoxes. Un tel mécanisme a une conséquence épidémiologique considérable, puisqu'il permet une transmission de pathogènes entre les troupeaux. Les virus de l'anémie infectieuse équine, de la fièvre porcine africaine, des fièvres à West Nile ou de la Vallée du Rift, sont connus pour être transmis par des stomoxes ; d'autres sont suspectés. Des rickettsies (Anaplasma, Coxiella), d'autres bactéries et des parasites (Trypanosoma spp., Besnoitia spp.) sont aussi transmis par les stomoxes. Enfin, les stomoxes sont aussi des hôtes intermédiaires de l'helminthe Habronema microstoma et pourraient être impliqués dans la transmission de certaines espèces d'Onchocerca et de Dirofilaria. En étant cosmopolite, Stomoxys calcitrans pourrait avoir un impact plus important qu'initialement imaginé sur la transmission de pathogènes aux animaux et aux humains.

This paper reviews the transmission modes ofTrypanosoma evansi. Its worldwide distribution is attributed to mechanical transmission. While the role of tabanids is clear, we raise questions on the relative role ofHaematobiasp. and the possible role ofStomoxyssp. in delayed transmission. A review of the available trypanocidal drugs and their efficacy in various host species is useful for understanding how they interact in disease epidemiology, which is complex. Although there are similarities with other mechanically transmitted trypanosomes,T. evansihas a more complex epidemiology due to the diversity of its hosts and vectors. The impact of clinical and subclinical disease is difficult to establish. A model was developed for buffaloes in the Philippines, which could be transferred to other places and livestock systems. SinceTrypanosoma evansiwas reported in humans, further research is required to investigate its zoonotic potential. Surra remains a potentially emerging disease that is a threat to Australia, Spain, and France. A number of questions about the disease have yet to be resolved. This brief review of the basic knowledge ofT. evansisuggests that there is renewed interest in the parasite, which is spreading and has a major economic impact.

Trypanosoma evansi is responsible for the most largely distributed animal trypanosomosis, affecting a wide range of wild and domestic animals. Its surveillance requires the implementation of standardized and reliable diagnostic tools. Although the development of polymerase chain reaction (PCR) tools has greatly improved our diagnostic capacity, factors affecting their sensitivity need to be acknowledged and accounted for in the interpretation of results. The targeted gene and the primer design have already been shown to greatly affect the sensitivity of a PCR, and the best-performing sets of primers have been previously identified. However, the sensitivity of the PCR is also largely influenced by the DNA extraction or sample preparation method. In this paper, we selected 6 DNA extraction or blood sample preparation methods representative of what would be used in a budget-constrained setting: phenol-chloroform, Chelex(®), Flexigen (Qiagen(®)) kit, Genekam(®) kit and two original protocols using sodium hydroxide. We studied the effects of the preparation method on the detection limit of the subsequent PCR. Our results show that the extraction method strongly affects the PCR sensitivity. The classical phenol-chloroform extraction method allowed for the PCR with the lowest detection limit. Some combinations of extraction method and primer set had detection limits that were not compatible with their use as a reliable diagnostic test, and would severely reduce the performance of a surveillance program. Therefore, we encourage laboratories to carefully select their sample preparation and PCR protocols, depending on the aimed sensitivity, cost, safety, time requirement and objectives.

Trypanosoma congolense type forêt a été identifié par PCR en France, chez un chien de retour du Sénégal. Cette publication décrit les divers aspects morphologiques du parasite sur des frottis sanguins colorés au Giemsa. Les formes fines (slender) et les corps latents représentent respectivement 30,4 % et 20,4 % des parasites. Quelques rosettes ont été observées (0,8 %). Le morphotype prédominant (48,4 %) est trapu (stumpy), proches des formes "montgomeryi", mais d'une largeur inhabituelle, avec un rapport largeur/longueur (rl/L) de 0,40-0,55, alors que celui des formes montgomeryi précédemment décrites est de l'ordre de 0,3. À notre connaissance, ceci est la première description de tels morphotypes de T. (Nannomonas). La forme du cytoplasme qui semble "tendu" par un flagelle en forme de "S" ou de "C" est également surprenante. Nous proposons de nommer ce morphotype "hyperpachymorphe", et de l'ajouter à la description de T. congolense type forêt. Ainsi, le sous-genre T. (Nannomonas) présenterait six morphotypes (du plus fin au plus large) : hyperleptomorphe (forme rodhaini, très long et mince avec flagelle libre); leptomorphe (forme simiae, mince avec flagelle libre), isomorphe (forme congolense, court généralement, sans flagelle libre), pachymorphe (forme montgomeryi, court et large (0,25 < rl/L < 0,34), sans flagelle libre), hyperpachymorphe (forme "hyper-montgomeryi", court et très trapu (0,35 < rl/L < 0,7), au cytoplasme clair et sans flagelle libre) et sphaeromorphe ou "corps latent" (globulaire).

In this study, we describe the first survey in Thailand of Trypanosoma theileri, a widespread and prevalent parasite of cattle that is transmitted by tabanid flies. Investigation of 210 bovine blood samples of Thai cattle from six farms by hematocrit centrifuge technique (HCT) revealed 14 samples with trypanosomes morphologically compatible to T. theileri. Additional animals were positive for T. theileri by PCR based on the Cathepsin L-like sequence (TthCATL-PCR) despite negative by HCT, indicating cryptic infections. Results revealed a prevalence of 26±15% (95% CI) of T. theileri infection. Additionally, 12 samples positive for T. theileri were detected in cattle from other 11 farms. From a total of 30 blood samples positive by HCT and/or PCR from 17 farms, seven were characterized to evaluate the genetic polymorphism of T. theileri through sequence analysis of PCR-amplified CATL DNA sequences. All CATL sequences of T. theileri from Thai cattle clustered with sequences of the previously described phylogenetic lineages TthI and TthII, supporting only two major lineages of T. theileri in cattle around the world. However, 11 of the 29 CATL sequences analyzed showed to be different, disclosing an unexpectedly large polymorphic genetic repertoire, with multiple genotypes of T. theileri not previously described in other countries circulating in Thai cattle.

Des enquetes parasitologiques et serologiques portant sur 1470 bovins ont ete realisees sur les sites de Korondougou et de Koutouba dans la region d’Odienne, zone a glossines, et sur les sites de Kasievogo et de Litio dans la region de Korhogo, zone presumee indemne en glossines. Les prevalences totales des infestations trypanosomiennes dans les regions d’Odienne et de Korhogo ont ete, respectivement, de 3,6 % et 2,4 %. Sur les sites de Korondougou, Koutouba, Kasievogo et Litio, elles ont ete respectivement de 1,4 %, 5,7 %, 2,7 % et 2,1 %. Des variations significatives de la prevalence, entre le sexe et les classes d’âge des bovins ont ete observees sur le site de Koutouba. Une seroprevalence de 43 % a ete observee dans chacune des deux regions. Sur les differents sites, elle a ete de 25,7 % a Korondougou, 60 % a Koutouba, 18,6 % a Kasievogo et de 72,3 % a Litio. Ce parametre est influence significativement par le phenotype, l’âge et le sexe, uniquement a Koutouba et Litio. Dans l’ensemble, cette etude a montre que les prevalences des infestations trypanosomiennes chez les bovins etaient faibles ( 5 %) a l’echelle des sites prospectes, contrairement aux seroprevalences obtenues ( 18 %). Mots-cles : Bovin, Trypanosomose, prevalence, seroprevalence, Cote d’Ivoire.

Here, two recent outbreaks of Trypanosoma evansi infection in mainland France and Spain associated with the importation of dromedary camels from the Canary Islands, are reported. The disease is well-known on the Archipelago since 1997 and many efforts have been made towards control and eventual eradication, but some areas still remain affected. Both mainland outbreaks were controlled by means of massive treatments and monthly serological, parasitological and molecular (PCR) evaluations carried out by Valencian Regional Animal Health laboratory and by CIRAD, Montpellier, respectively. Possible causes for the persistence of the parasite in a small area of the Canaries are discussed. T. evansi must be included among the animal health conditions for international trade within the European Union as well as many other countries. Moreover, procedures including diagnosis, curative or preventive treatment and quarantine should be established to insure the status of the animals moving from a country to another.

To face the worldwide threat of Surra caused by Trypanosoma evansi, international organizations have stressed the need to evaluate and standardize diagnostic tools. PCR detection of T. evansi has known a great expansion during the last 20 years, but primer sets are often insufficiently assessed and compared. In this work, we compared the performances of six primer pairs–TBR1/2 ( Masiga et al., 1992 ), ESAG6/7 ( Holland et al., 2001a , Holland et al., 2001b ), TEPAN1/2 ( Panyim et al., 1993 ), pMUTEC F/R ( Wuyts et al., 1994 ), TRYP1 R/S ( Desquesnes et al., 2001 ) and TRYP4 R/S (Desquesnes et al., unpublished)–tested with purified T. evansi DNA serial dilutions, T. evansi-infected rat blood serial dilutions and Thai dairy cattle samples. TBR1/2 primer set was able to detect 0.01 pg of purified DNA, and a parasitaemia below one parasite per ml in rat blood. They presented the highest sensitivity in cattle samples as well as a high specificity, without non-specific products nor false positive reactions out of 84 negative cattle samples tested. ESAG6/7 showed equivalent results with purified DNA and rat samples but presented non-specific products with Thai dairy cattle samples, leading to non interpretable results. TEPAN1/2 was not able to detect less than 0.1 pg of purified DNA or 50 trypanosomes/ml in rat blood. In cattle, TEPAN1/2 primers detected only 36% of the positives detected by TBR1/2. Given the parasitemic level detected, pMUTEC F/R, TRYP1 R/S and TRYP4 R/S were not more sensitive than classical microscopic examination of the buffy coat. TBR1/2, TEPAN1/2, pMUTEC F/R and TRYP4 R/S did not cross-reacted with Babesia sp., Trypanosoma theileri and Anaplasma marginale. TBR1/2 was the most sensitive primer set to detect T. evansi in purified DNA, rodent blood and cattle blood, and did not show cross reaction with the other pathogens tested: it should be therefore preferred for epidemiological surveys. These results confirmed that TBR1/2 primers remain the reference for the detection of Trypanozoon DNA and should therefore be included in subsequent evaluations of new diagnosis tools based on DNA detection.

Trypanosoma evansi is generally considered a mild pathogen in bovines. However, in Asia, acute and chronic signs have been observed in cattle, with high levels of parasitaemia, abortion and death. Investigations in Asian cattle are needed to better understand this epidemiological situation. To generate comparable data at a regional level, development and standardization of an antibody-enzyme linked immunosorbent assay for T. evansi (ELISA/ T. evansi ) was initiated and applied in an epidemiological survey carried out in dairy cattle in Thailand. A batch of 1979 samples was collected from dairy farms located throughout the country's four regions. Soluble T. evansi antigens initially produced in France were also produced in Thailand for comparison and technology transfer. Screening of 500 samples allowed us to identify reference samples and to determine the cut-off value of the ELISA. Seropositive animals – some of them confirmed by PCR – were found in the four regions, in 12 out of 13 provinces, in 22 out of 31 districts, in 56 farms out of 222 (25%, 95%CI ± 6%) and in 163 animals out of 1979 (8.2, 95%CI ± 1.2%). Estimated seroprevalence in 35 farms ranged between 1% and 30%, and in 21 farms it was >30%. Approximately 25% of survey cattle were exposed to the infection, in various situations. A sub-sample of 160 sera was tested on both antigens. Wilcoxon's ( Z = 1.24; p = 0.22) and McNemars's tests (CHI2 = 3.55; p = 0.09) did not show any significant differences, showing that the locally produced antigen is suitable for further evaluation in the surrounding countries. Use of this standardized serological method will broaden knowledge of the prevalence and impact of the disease at the regional level in South-East Asia. Further validation of this ELISA will be necessary in other host species such as buffalo, horse and pig.

Erratum paru dans Veterinary parasitology (2009) 165 (1-2) p. 185 http://dx.doi.org/10.1016/j.vetpar.2009.03.058. An outbreak of trypanosomosis was observed for the first time in metropolitan France in October 2006, when five camels were proved to be infected by Trypanosoma evansi using parasitological methods. The parasite was isolated and used to produce a soluble antigen for antibody-enzyme linked immunosorbent assay (ELISA) in a protocol derived from a method previously developed for sheep and humans but using protein A conjugate. The animals were treated on three instances, alternatively with melarsomine hydrochloride and quinapyramine and followed up on a monthly basis for 2 years with various diagnostic techniques including parasitological, serological and DNA-based methods. Initially, five animals were detected as being positive using ELISA with 83.3% concordance to parasitological tests. Immediately after the first treatment, parasites and DNA disappeared in all animals; antibody levels decreased regularly until ELISA became negative 3-4 months later. Ten months after the first treatment, parasites and antibodies were detected again in one of the camels previously found to be infected. A retrospective study indicated that the weight of this animal had been underestimated; consequently, it had received underdosages of both trypanocides. However, since hypotheses of re-infection or relapse could not be fully substantiated, it is not known whether the ELISA results for this animal were true- or false-negative over a 7-month period. The study confirmed the value of this ELISA using protein A conjugate to detect antibodies directed against T. evansi in camels and the need to use several diagnostic techniques to optimize detection of infected animals. A warning is raised on surra, a potentially emerging disease in Europe.

The biology, epidemiology, pathogenesis, diagnostic techniques, and history of the introduction of Trypanosoma (Duttonella) vivax in the New World are reviewed. The two main immunological responses of trypanosome-infected animals - antibody production and immunodepression - are discussed in the context of how these responses play a role in disease tolerance or susceptibility. Isolation and purification of T. vivax are briefly discussed. The recent reports of bovine trypanosomiasis diagnosed in cattle on farms located in the Pantanal region of the states of Mato Grosso do Sul and Mato Grosso, Brazil, are also discussed.

The first outbreak of trypanosomosis caused by Trypanosoma evansi in camels in France was reported on a farm in the Aveyron Department. Five camels were imported from the Canary Islands to the farm in early July 2006, and trypanosomes were observed on a stained blood smear from one of them, which died in October. On further investigations, trypanosomes were observed in the blood of five camels, three of them indigenous to the farm and two that had been imported. On the basis of microscopical examination (morphological criteria and measurements) and serological results based on the card agglutination T evansi test and PCR typing, the parasites were identified as T evansi. After treatment with melarsomine, the infected camels rapidly became negative by parasitological tests and were negative two to four months later by serological tests. The parasite was probably transmitted by tabanids and Stomoxys calcitrans, which were abundant in July to September 2006. No parasites were observed in other animals on the farm or on neighbouring farms, but some of the sheep on these farms were positive by PCR or serology.

International audience; In Burkina Faso, African Animal Trypanosomosis (AAT) is still a major hindrance to cattle breeding, especially in the Mouhoun river basin, which was identified as a priority area for tsetse control. The attempt of the present work was to assess the abundance of tsetse flies and AAT risk using remote sensing coupled to field environmental data, along a Mouhoun river section of 234 km long, harbouring an open riverine forest where G. tachinoides Westwood is the predominant tsetse species. The water course was classified into three epidemiological landscapes, corresponding to a "disturbed", "natural" and finally "border" vegetal formation at the interface of the two formers. Using the mean number of infected flies by trap and by day as a risk indicator, the border landscape was found to be 5.4 (1.3-12.0) and 15.8 (4.7-41.6) times more risky than the natural and disturbed ones respectively. These results led to propose that a campaign against tsetse, undertaken by a development project called PAEOB (Projet d'Appui à l'Élevage dans l'Ouest du Burkina Faso), should be focussed on only 34% of the hydrographic network.

A longitudinal epidemiological survey of bovine trypanosomosis and its vectors was carried out in the Volta river basin of Northern Ghana to determine the relationship between cattle management and the incidence of bovine trypanosomosis. Two groups of sentinel cattle under different systems of management, classified as "fully-sedentary" and "partially-sedentary" (depending on the type of management) were followed over a 1-year period starting from March 2003 onwards. Cattle were screened at intervals of 3 months using the huffy coat technique (BCT). Buffy coat specimen from animals that were positive for the BCT and those that were negative, but with a packed cell volume (PCV) of less than 21% were further tested using the polymerase chain reaction (PCR). Plasma from all animals were tested for antibody using the indirect antibody enzyme-linked immunosorbent assay (ELISA). Trypanosomosis challenge was determined in tandem with the epidemiological survey with watering sites of sentinel cattle being the foci of interest. The parasitological prevalence at the start of the survey was higher in the fully-sedentary group (9%) than in the partially-sedentary group (3%). In subsequent visits, however, the parasitological incidence was consistently higher in the partially-sedentary group than in the fully-sedentary group. The mean seroprevalence (ELISA) of both groups increased from 3% in March to 54% in December. Statistical analysis of the serological results using a random effect logistic regression, showed a significant difference in incidence of bovine trypanosomosis between the two groups. There was also a significant effect of time. The influence of cattle herding on host-vector-parasite interface and its consequence on the incidence of trypanosomosis are discussed. (c) 2004 Elsevier B.V. All rights reserved.

The epidemiology of bovine trypanosomosis was investigated in two districts (Savelugu and West Mamprusi) of Northern Ghana with different land use and environmental characteristics. The land use intensity and environmental change was suspected to be higher in the Savelugu District. A cross-sectional entomological survey conducted along the White Volta river and its tributaries confirmed the presence of only Glossina palpalis gambiensis and G. tachinoides. The challenge index as measured by the product of tsetse density and tsetse infection rate was much higher in the West Mamprusi (19.6) than in the Savelugu district (4.7). A total of 1013 cattle (508 in Savelugu and 505 in West Mamprusi) were bled from a random selection of 16 villages in the Savelugu District and 13 villages in the West Mamprusi District. Blood samples were examined for trypanosomes by the buffy coat technique (BCT). Blood samples that were positive in the BCT or negative in the BCT but with packed cell volume (PCV) values below 21 were further tested with a polymerise chain reaction for trypanosomal DNA. Plasma samples of all cattle were serologically tested with an indirect ELISA for trypanosomal antibodies. The parasitological and serological prevalence of bovine trypanosomoses was significantly higher in West Mamprusi (16 and 53%, respectively) than in Savelugu District (8 and 24%, respectively). An evaluation of animal health at the village herd level, using PCV as an index of anaemia, provided various epidemiological scenarios prevalent in the entire study area. © 2004 Elsevier B.V. All rights reserved.

The role of mechanical vectors in the transmission of African livestock trypanosomes has always been controversial relative to tsetse flies, their cyclical vectors. An experiment was carried out in Burkina Faso to demonstrate mechanical transmission of Trypanosoma vivax by one of the most common tabanids in Africa: Atylotus agrestis. Eight heifers (crossbred zebu × Baoule), free of trypanosome infection, were kept in a corral covered by a mosquito net, together with two heifers infected experimentally with a local stock of T. vivax. On average, 324 A. agrestis, freshly captured with Nzi traps, were introduced daily over 20 days. Parasitological, PCR and serological examinations were carried out regularly to assess infections and levels of parasitaemia. Microscopic examination of buffy-coats indicated that five of the eight receiver-heifers were infected on days 8, 13, 32, 41, and 48. PCR results indicated that these five heifers were already infected by day 13. Mechanical transmission of T. vivax by A. agrestis was demonstrated unequivocally, at a high rate (63% in 13–20 days). Conditions of transmission in this experiment are discussed in terms of natural rates of challenge. The importance of tabanids as mechanical vectors of T. vivax should be re-considered, in light of these results. Creation of tsetse free zones in Africa will generally lead to the disappearance of T. congolense, T. brucei, and most often T. vivax as well; however, in areas where T. vivax can be mechanically transmitted, clearance of tsetse may not be sufficient to eradicate livestock trypanosomosis. Index Descriptor and Abbreviations: Trypanosoma vivax (Trypanosomatidae) is a pathogenic trypanosome found in wild and domestic herbivores, principally in cattle (Bos taurus, Bos indicus, and crossbreeds), in South America and Africa. In Africa it is principally cyclically transmitted by tsetse flies (Diptera). In America it is only mechanically transmitted by tabanids (Tabanidae). The present paper describes the mechanical transmission of T. vivax in cattle by one of the most common African tabanids: A. agrestis. The main conclusion is that not only tsetse can transmit T. vivax in Africa, consequently the epidemiology of trypanosomosis in cattle is also tabanid dependant, and, the eradication of tsetse flies will not necessarily lead to eradication of T. vivax. ADT, Apparent density of insects per trap per day (mean number of insects caught in one type of trap per 24 h of trapping); PATTEC, Pan African Tsetse and Trypanosomiasis Eradication Campaign; RHD, rate of hematocrit decrease; D, day; ELISA, enzyme linked immunosorbent assay; NS, not significant (in terms of statistics); PCR, polymerase chain reaction

The pathology of African bovine trypanosomosis was compared in Zebu cattle subcutaneously inoculated with three clones of trypanosomes corresponding to the three genetically distinct types of Trypanosoma congolense; savannah-type, west African riverine/forest-type and kilifi-type. All inoculated animals became parasitaemic between 7 and 11 days post-infection (dpi). The savannah-type showed consistently higher levels of parasitaemia and lower packed red cell volume percentages and leukocyte counts than the other two types. The syndrome was also more severe in the savannah-type and led inexorably to death between 29 and 54 dpi while animals with the forest or the kilifi-types recovered from earlier symptoms and haematological alterations after 3 months of infection. By the end of the experiment, the animals self-cured from the forest-type infection and the kilifi-type passed under control. The results of the present study indicated clear difference in pathogenicity between the three types of T. congolense; the savannah-type was virulent while the forest-type was of low pathogenicity and the kilifi-type was non-pathogenic.

The diagnostic performance of a polymerase chain reaction assay (PCR) for monitoring the effectiveness of aceturate diminazene treatment was compared with those of an antibody-detection ELISA test and the buffy-coat technique using sheep experimentally infected with either savannahtype or forest-type Trypanosoma congolense or T. vivax. Within the period of infection, the PCR using specific savannah-type T. congolense primers showed a significant higher diagnostic sensitivity (p < 0.05) than the buffy-coat technique. Both techniques gave closed results for detecting forest-type T. congolense or T. vivax infections. Following trypanocidal treatment, the PCR showed that specific product disappeared definitively 1 or 2 days later in animals in which a decrease of the antibody level and a significant improvement of the red packed cell volume were observed. The occurrence of relapse infection was detected by the PCR in one animal infected by T. vivax on day 19 post-treatment and confirmed by the persistence and increasing antibody level whereas the buffy-coat technique detected parasites 42 days later. Then, the PCR signals remained positive on several occasions while parasitaemia was detected only two times. The application of PCR combined with the antibody detection appeared to provide a useful tool as compared to the buffy-coat technique for monitoring the effectiveness of trypanocidal treatment.

The two classical forms of human trypanosomoses are sleeping sickness due to Trypanosoma brucei gambiense or T. brucei rhodesiense, and Chagas disease due to T. cruzi. However, a number of atypical human infections caused by other T. species (or sub-species) have been reported, namely due to T. brucei brucei, T. vivax, T. congolense, T. evansi, T. lewisi, and T. lewisi-like. These cases are reviewed here. Some infections were transient in nature, while others required treatments that were successful in most cases, although two cases were fatal. A recent case of infection due to T. evansi was related to a lack of apolipoprotein L-I, but T. lewisi infections were not related to immunosuppression or specific human genetic profiles. Out of 19 patients, eight were confirmed between 1974 and 2010, thanks to improved molecular techniques. However, the number of cases of atypical human trypanosomoses might be underestimated. Thus, improvement, evaluation of new diagnostic tests, and field investigations are required for detection and confirmation of these atypical cases. © 2013 Truc et al., SCOPUS: ar.j, info:eu-repo/semantics/published

Une synthèse des connaissances sur l'écologie, l'importance épidémiologique et le contrôle des stomoxes à l'île de la Réunion est présentée. L'intensification de l'élevage bovin dans ce département a créé des conditions optimales pour la pullulation de ces insectes hématophages. Les stomoxes sont non seulement une nuisance directe par la spoliation sanguine et le stress lié au harcèlement des animaux, mais aussi et surtout une nuisance indirecte, en tant que vecteurs mécaniques de nombreuses maladies animales. À la Réunion, le principal rôle pathogène des stomoxes est la transmission de l'anaplasmose bovine, qui est une contrainte sanitaire majeure pour l'élevage de races européennes. Des études récentes ont permis de mieux appréhender leur distribution, leur dynamique et leur structure de populations, mais leur contrôle, organisé par le Groupement de Défense Sanitaire, reste problématique. Les, essais mis en place depuis 1994, ont donné des résultats variables ; les perspectives sont discutées. De nombreuses pistes de recherche sont explorées pour mieux comprendre leur rôle dans la transmission mécanique immédiate ou différée des agents pathogènes. L'amélioration de leur contrôle pourra être assurée en mettant en place une politique de gestion intégrée à l'échelle de l'île, basée sur une meilleure prise en compte des connaissances sur leur écologie et sur l'intégration de nouvelles méthodes de lutte (pédiluves, champignons entomopathogènes, moustiquaires imprégnées).