Modelling habitat suitability of the invasive tick Rhipicephalus microplus in West Africa.

Ticks have medical and economic importance due to their ability to transmit pathogens to humans and animals. In tropical and sub-tropical countries, tick-borne diseases (TBD) are among the most important diseases affecting livestock and humans. The fast spread of ticks and TBD requires a quick development and application of efficient prevention and/or control programs. Therefore, prior investigations on TBD and related vectors epidemiology, for instance, through accurate epidemiological models, are mandatory. This study aims to develop models to forecast suitable habitat for Rhipicephalus microplus distribution in West Africa. Tick occurrences were assembled from 10 different studies carried out in six West African countries in the past decade. Six statistical models (maximum entropy in a single model and generalised linear model, generalised additive model, random forest, boosted regression tree and support vector machine model in an ensemble model) were applied and compared to predict the habitat suitability of R. microplus distribution in West Africa. Each model was evaluated with the area under the receiver operating characteristic curve (AUC), the true skill statistic (TSS) and the Boyce index (BI). The selected models had good performance according to their AUC (above .8), TSS (above .7) and BI (above .8). Temperature played a key role in MaxEnt model, whereas normalised difference vegetation index (NDVI) was the most important variable in the ensemble model. The model predictions showed coastal countries of West Africa as more suitable for R. microplus. However, some Sahelian areas seems also favourable. We stress the importance of vector surveillance and control in countries that have not yet detected R. microplus but are in the areas predicted to host suitable habitat. Indeed, awareness-raising and training of different stakeholders must be reinforced for better prevention and control of this tick in these different countries according to their status.

Models for Studying the Distribution of Ticks and Tick-Borne Diseases in Animals: A Systematic Review and a Meta-Analysis with a Focus on Africa.

Ticks and tick-borne diseases (TTBD) are constraints to the development of livestock and induce potential human health problems. The worldwide distribution of ticks is not homogenous. Some places are ecologically suitable for ticks but they are not introduced in these areas yet. The absence or low density of hosts is a factor affecting the dissemination of the parasite. To understand the process of introduction and spread of TTBD in different areas, and forecast their presence, scientists developed different models (e.g., predictive models and explicative models). This study aimed to identify models developed by researchers to analyze the TTBD distribution and to assess the performance of these various models with a meta-analysis. A literature search was implemented with PRISMA protocol in two online databases (Scopus and PubMed). The selected articles were classified according to country, type of models and the objective of the modeling. Sensitivity, specificity and accuracy available data of these models were used to evaluate their performance using a meta-analysis. One hundred studies were identified in which seven tick genera were modeled, with Ixodes the most frequently modeled. Additionally, 13 genera of tick-borne pathogens were also modeled, with Borrelia the most frequently modeled. Twenty-three different models were identified and the most frequently used are the generalized linear model representing 26.67% and the maximum entropy model representing 24.17%. A focus on TTBD modeling in Africa showed that, respectively, genus Rhipicephalus and Theileria parva were the most modeled. A meta-analysis on the quality of 20 models revealed that maximum entropy, linear discriminant analysis, and the ecological niche factor analysis models had, respectively, the highest sensitivity, specificity, and area under the curve effect size among all the selected models. Modeling TTBD is highly relevant for predicting their distribution and preventing their adverse effect on animal and human health and the economy. Related results of such analyses are useful to build prevention and/or control programs by veterinary and public health authorities.

Efficacy of two commercial synthetic pyrethroids (cypermethrin and deltamethrin) on Amblyomma variegatum and Rhipicephalus microplus strains of the south-western region of Burkina Faso.

Since 2011, period of the livestock invasion by the cattle tick Rhipicephalus microplus in Burkina Faso (BF), tick-control problems were exacerbated. Based on farmer's reports, most commonly used commercial acaricides were found to be ineffective in Western South part of the country. To investigate the occurrence and extent of such acaricidal ineffectiveness, we performed the standardized larval packet test (LPT) with commercial deltamethrin (vectocid) and cypermethrin (cypertop), on two cattle tick species, the native Amblyomma variegatum and the invasive R. microplus. The resistance ratios (RR) were computed with susceptible Hounde strain of Rhipicephalus geigyi as reference. The R. microplus population showed resistance to the two acaricides tested with the highest lethal concentration (LC) values, and different resistance ratios higher than 4 (deltamethrin: RR50 = 28.18 and RR90 = 32.41; cypermethrin: RR50 = 8.79 and RR90 = 23.15). In the contrary, A. variegatum population was found to be highly susceptible to acaricides tested with low lethal concentrations and resistance ratio values (deltamethrin: RR50 = 0.5 and RR90 = 0.48; cypermethrin: RR50 = 0.68 and RR90 = 0.79). These data demonstrate high synthetic pyrethroid resistance in R. microplus strain, leading to conclude that the acaricide ineffectiveness in tick populations control remains a concern in BF.

Cross border transhumance involvement in ticks and tick-borne pathogens dissemination and first evidence of Anaplasma centrale in Burkina Faso.

In West Africa, cross-border transhumance, also called seasonal migration, is known to be a very important animal production strategy, as it involves about 70 to 90% of cattle. In spite of the cattle movements, some strategic areas of transhumance remain poorly explored regarding ticks and their associated pathogens investigations. The purpose of this study is to evaluate the involvement of transhumance in the spread of cattle ticks and associated pathogens in Burkina Faso (BF) and Benin (BN), in a context of speedy invasion of West African livestock by Rhipicephalus microplus. A longitudinal survey was performed on 210 cattle from BF, monitored for ticks and tick-borne pathogens (TBP) during one seasonal transhumance. The first sampling coded "T0BF" took place in eastern BF, at the transhumance departure. A second sampling "T1BN" was carried out in northern BN, the transhumance arrival zone. A third sampling "T2BF" was done at the return of cattle in eastern BF. Ticks were morphologically identified and TBP detected with reverse line blot hybridization (RLB) assay. A total of 1027 ticks (7 species), 1006 ticks (11 species) and 1211 ticks (9 species) were respectively found at T0BF, T1BN and T2BF. Some species were collected at the three times of sampling without any significant difference in their relative abundances. However, other tick species appeared only at T1BN and/or T2BF. The TBP species found at the three points surveyed were Theileria annulata, Theileria mutans, Theileria velifera, Babesia bigemina and Anaplasma marginale. The most prevalent was T. mutans with 166/210 (79%), 159/210 (75.7%) and 78/210 (37%) cattle positive respectively at T0BF, T1BN and T2BF. Anaplasma centrale was evidenced with 0.5% and 0.9% respectively at T0BF and T2BF. To our knowledge, this represents its first report in the study area. Overall, the TBP prevalences were significantly lower at T2BF, highlighting the effect of tick populations changes induced by transhumance combined with the seasonal variation influence.

Cattle ticks and associated tick-borne pathogens in Burkina Faso and Benin: Apparent northern spread of Rhipicephalus microplus in Benin and first evidence of Theileria velifera and Theileria annulata.

Babesiosis, theileriosis, anaplasmosis, and heartwater are tick-borne diseases that threaten livestock production in sub-Saharan Africa including Burkina Faso and Benin. For over a decade, these two bordering countries have been facing an invasion of the livestock by the tick Rhipicephalus microplus, a major vector for babesiosis, accidentally introduced in Benin in 2004. The molecular identification of tick-borne pathogens in this border area is of particular interest due to animals seasonal migration between the two countries. In this survey, epidemiological features of ticks and tick-borne pathogens in cattle were investigated to compare the eastern Burkina Faso, corresponding to a seasonal migration departure zone, and the northern Benin, which represents a seasonal migration arrival zone. Ticks and peripheral blood were collected from a total of 946 cattle in the two areas. Ticks were morphologically identified and the DNA samples from bovine blood and ticks were analysed by Reverse Line Blot (RLB) hybridization process. A total of 2856 ticks were collected on 490 cattle in Burkina Faso, eight tick species were identified, while 3583 ticks were collected on 456 cattle in North Benin with nine tick species identified. The invasive tick, R. microplus was not found in eastern Burkina Faso, but its spread farthest north in Benin is reported. Six tick-borne pathogen species were found in cattle blood both in eastern Burkina Faso and in northern Benin. Ranked in decreasing order of overall prevalences, they are: Theileria mutans (91.1%), Theileria velifera (77.8%), Babesia bigemina (10.9%), Anaplasma marginale (4.2%), Babesia bovis (3.3%), and Theileria annulata (1.8%). To the best of our knowledge, this survey represents the first report of T. velifera and T. annulata in the region. Overall, the TBP prevalences were significantly higher in northern Benin than in eastern Burkina Faso, indicating a higher parasitological risk in this area.

First tick and tick damage perception survey among sedentary and transhumant pastoralists in Burkina Faso and Benin.

BACKGROUND: Transhumance, a main ancestral animal production strategy of the West African Countries (WAC), can favour the spread of vectors and vector-borne diseases within and/or across countries. Transhumance has been implicated in such spread as well as that of related tick-borne diseases (TBD). METHODS AND PRINCIPAL FINDINGS: Using a questionnaire survey and statistical modelling, this study explores the perception of herders about ticks and TBD in cattle, their practices in tick control and the social groups involved in cattle farming in eastern Burkina Faso (46 random herds) and in the northern Benin (44 random herds). Results show that most of the herders (79%) are from the Fulani social group. The principal and secondary activities of herders are respectively cattle farming and agriculture. The mean age of pastoralists is between 40 and 50 years depending on the province of origin and 60% of the surveyed herds practice internal or transboundary transhumance. Herders have a clear knowledge of different genus of ticks except the genus Rhipicephalus. Their knowledge of TBD is very limited. These results also reveal that herders in Benin use less acaricides treatment calendar compared with those in Burkina Faso. Transhumant pastoralists (i.e. transhumant cattle farmers) plan more acaricide treatment and have more cows with lost teats (i.e. tick damage) than the sedentary ones. In addition, amitraz appears to be the main acaricide compound used by herders for tick control (68%) but its use is inappropriate and its source is frequently the unregulated market. CONCLUSIONS AND SIGNIFICANCE: All of these findings can induce acaricide resistance especially as the inefficacy of amitraz against Rhipicephalus (Boophilus) microplus has already been reported in previous studies. Such results would help to elaborate suitable strategies of control and prevention of ticks and TBD in Burkina Faso and Benin.

Insights on foodborne zoonotic trematodes in freshwater snails in North and Central Vietnam.

Foodborne zoonotic trematode (FZT) infections are common neglected tropical diseases in Southeast Asia. Their complicated life cycles involve freshwater snails as intermediate hosts. A cross-sectional study was conducted in Yen Bai and Thanh Hoa provinces in North and Central Vietnam, to investigate the diversity of cercariae of potential FZT and to construct the phylogenetic relationship of trematode cercariae based on the Internal Transcribed Spacer 2 (ITS2) region. Among 17 snail species collected from various habitats, 13 were infected by 10 cercarial groups among which parapleurolophocercous, pleurolophocercous, and echinostome cercariae were of zoonotic importance. The monophyletic tree separated cercarial sequences into different groups following the description of the cercariae families in which Haplorchidae, Opisthorchiidae, Echinochasmidae, and Echinostomatidae are important families of FZT. The overall prevalence was different among snail species and habitats and showed a seasonal trend. Parapleurolophocercous and echinostome cercariae emerged as the most common cercariae in snails in Yen Bai, while monostome, echinostome, and megalura cercariae were most common in Thanh Hoa. Using a molecular approach, we identified Parafossarulus striatulus as the first intermediate snail host of Clonorchis sinensis in Thac Ba Lake. Melanoides tuberculata and Bithynia fuchsiana were we identified preferred intermediate snail hosts of a diverse range of trematode species including intestinal flukes (i.e., Haplorchis pumilio and Echinochasmus japonicus) in Yen Bai and Thanh Hoa, respectively.

First digital characterization of the transhumance corridors through Benin used by cattle herds from Burkina Faso and associated risk scoring regarding the invasion of Rhipicephalus (Boophilus) microplus.

BACKGROUND: Livestock plays a key role in the macro-economy of West Africa and provides livelihoods for millions of people. The main cattle rearing strategy in West Africa is pastoralism, including transhumance, that is a seasonal migration of cattle with their herders. This adaptive strategy aims to optimize livestock access to water and pastures. However, it can favour pathogens and vectors transboundary spread. The aim of this study was to highlight firstly, the corridors and grazing areas used by Burkina Faso transhumant cattle herds going to Benin; secondly, the characteristics of departure and arrival areas of transhumance; and thirdly, the risk score related to introduction and spread of the invasive tick species, Rhipicephalus (Boophilus) microplus, in free areas. METHODS AND PRINCIPAL FINDINGS: Therefore, GPS devices were given to 27 herders to monitor a full transhumance season between East Burkina Faso and North Benin. The analysis of 14,966 spots generated by the GPS devices revealed four main corridors and five main grazing areas used by cattle herds during transhumance. Statistical analysis of normalized difference vegetation index (NDVI), rainfall and temperature data, highlighted significant differences between departure and arrival areas. NDVI and rainfall are significantly higher in Benin than Burkina Faso, whereas temperature is significantly lower. Additionally, using biotic and abiotic parameters, a risk scoring was developed to predict the presence of Rhipicephalus (Boophilus) microplus at municipality level. CONCLUSIONS AND SIGNIFICANCE: The better vegetation, temperature and rainfall conditions during the dry seasons in Benin attract cattle herds from Burkina Faso. The invasiveness and adaptability of Rhipicephalus (Boophilus) microplus added to the frequent stays of transhumant herds in infested areas suggest its potential introduction and establishment in free areas soon. Moreover, frequent intrusions of the transhumant cattle in the wildlife reserves are another risk of vectors and pathogen exchange between domestic and wild animals.

An expert opinion assessment of blood-feeding arthropods based on their capacity to transmit African swine fever virus in Metropolitan France.

To deal with the limited literature data on the vectorial capacity of blood-feeding arthropods (BFAs) and their role in the transmission of African swine fever virus (ASFV) in Metropolitan France, a dedicated working group of the French Agency for Food, Environmental and Occupational Health & Safety performed an expert knowledge elicitation. In total, 15 different BFAs were selected as potential vectors by the ad hoc working group involved. Ten criteria were considered to define the vectorial capacity: vectorial competence, current abundance, expected temporal abundance, spatial distribution, longevity, biting rate, active dispersal capacity, trophic preferences for Suidae, probability of contact with domestic pigs and probability of contact with wild boar. Fourteen experts participated to the elicitation. For each BFA, experts proposed a score (between 0 and 3) for each of the above criteria with an index of uncertainty (between 1 and 4). Overall, all experts gave a weight for all criteria (by distributing 100 marbles). A global weighted sum of score per BFA was calculated permitting to rank the different BFAs in decreasing order. Finally, a regression tree analysis was used to group those BFAs with comparable likelihood to play a role in ASF transmission. Out of the ten considered criteria, the experts indicated vectorial competence, abundance and biting rate as the most important criteria. In the context of Metropolitan France, the stable fly (Stomoxys calcitrans) was ranked as the most probable BFA to be a vector of ASFV, followed by lice (Haematopinus suis), mosquitoes (Aedes, Culex and Anopheles), Culicoides and Tabanidea. Since scientific knowledge on their vectorial competence for ASF is scarce and associated uncertainty on expert elicitation moderate to high, more studies are however requested to investigate the potential vector role of these BFAs could have in ASFV spread, starting with Stomoxys calcitrans.

Mechanical transmission of African swine fever virus by Stomoxys calcitrans: Insights from a mechanistic model.

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 (<5 flies per pig), stable flies would play a minor role in the spread of ASFV, as they are expected to be responsible for around 10% of transmission events. However, with abundances of 20 and 50 stable flies per pig, the vector-borne transmission would likely be responsible for almost 30% and 50% of transmission events, respectively. In these situations, time to reach a pig mortality of 10% would be reduced by around 26% and 40%, respectively. The sensitivity analysis emphasized that the expected relative contribution of stable flies was strongly dependent on the volume of blood they regurgitated and the infectious dose for pigs. This study identified crucial knowledge gaps that need to be filled in order to assess more precisely the potential contribution of stable flies to the spread of ASFV, including a quantitative description of the populations of haematophagous arthropods that could be found in pig farms, a better understanding of blood-feeding behaviours of stable flies and the quantification of the probability that stable flies partially fed with infectious blood transmit the virus to a susceptible pig during a subsequent blood-feeding attempt.

Prevalence and risk factors associated with Clonorchis sinensis infections in rural communities in northern Vietnam.

BACKGROUND: Clonorchiasis, caused by the fish-borne trematode Clonorchis sinensis, is a neglected tropical disease and a public health issue in endemic countries. In Vietnam, an in-depth analysis of risk factors for the condition is missing up to now. This study aimed to determine the prevalence of C. sinensis infection and associated risk factors in rural communities in northern Vietnam. METHODOLOGY/PRINCIPAL FINDINGS: A cross-sectional survey was conducted in 4 communes in Yen Bai and Thanh Hoa provinces where clonorchiasis is known to be present and raw fish consumption is a common. Using a simple random sampling approach, stool was collected from 841 participants over 6 years old for coprological examination, and a questionnaire measured knowledge, attitudes, and practices with regard to clonorchiasis in 757 participants over 15 years old. Univariable and multivariable logistic regression models were run to identify risk factors for infection with C. sinensis. The overall prevalence of C. sinensis infection was 40.4%, with commune prevalences ranging between 26.5% and 53.3%. In the final model, males were significantly more likely to be infected with C. sinensis (OR 2.00; 95% CI 1.31-3.05). Recent (i.e. last year) consumption of raw fish (OR 8.00, 95% CI 4.78-13.36), low education level (OR 5.57; 95% CI 2.37-13.07), lack of treatment (OR 1.82, 95% CI 1.15-2.89), being between 19 to 39 years old (OR 6.46; 95% CI 1.25-33.37), and the presence of an unhygienic toilet (OR 2.74, 95% CI 1.53-4.92) were significantly associated with C. sinensis infection. CONCLUSION/SIGNIFICANCE: This study demonstrated a high prevalence of C. sinensis infection in rural communities in northern Vietnam. Thus, control measures including, mass drug administration for those communes should be applied to reduce the prevalence. Moreover, specific health education activities should be developed for risk groups in C. sinensis endemic areas.

Putative Role of Arthropod Vectors in African Swine Fever Virus Transmission in Relation to Their Bio-Ecological Properties.

African swine fever (ASF) is one of the most important diseases in Suidae due to its significant health and socioeconomic consequences and represents a major threat to the European pig industry, especially in the absence of any available treatment or vaccine. In fact, with its high mortality rate and the subsequent trade restrictions imposed on affected countries, ASF can dramatically disrupt the pig industry in afflicted countries. In September 2018, ASF was unexpectedly identified in wild boars from southern Belgium in the province of Luxembourg, not far from the Franco-Belgian border. The French authorities rapidly commissioned an expert opinion on the risk of ASF introduction and dissemination into metropolitan France. In Europe, the main transmission routes of the virus comprise direct contact between infected and susceptible animals and indirect transmission through contaminated material or feed. However, the seasonality of the disease in some pig farms in Baltic countries, including outbreaks in farms with high biosecurity levels, have led to questions on the possible involvement of arthropods in the transmission of the virus. This review explores the current body of knowledge on the most common arthropod families present in metropolitan France. We examine their potential role in spreading ASF-by active biological or mechanical transmission or by passive transport or ingestion-in relation to their bio-ecological properties. It also highlights the existence of significant gaps in our knowledge on vector ecology in domestic and wild boar environments and in vector competence for ASFV transmission. Filling these gaps is essential to further understanding ASF transmission in order to thus implement appropriate management measures.

Detection of Two Species of the Genus Parapoxvirus (Bovine Papular Stomatitis Virus and Pseudocowpox Virus) in Ticks Infesting Cattle in Burkina Faso.

The molecular identification of arboviruses in West Africa is of particular interest, due to their zoonotic potential in a population living in close contact with livestock, and in a region where the livestock migration across borders raises the risk of diseases infection and dissemination. The aim of the study was the screening of potential circulating arboviruses and the assessment of their zoonotic implications. Therefore, ticks were collected on cattle located in three provinces of eastern Burkina Faso. Tick pools were tested using a panel of genus-specific real-time assays targeting conserved regions of parapoxvirus, orthopoxvirus, flavivirus and phlebovirus. On the 26 farms visited, a total of 663 ticks were collected. Four genera and six tick species were morphologically identified, with Amblyomma variegatum and Hyalomma spp. being the most represented species. No arboviruses were found. However, this study highlights the presence of pseudocowpox virus (8.2%) and bovine papular stomatitis virus (5.8%) among the positive tick pools. BPSV positive ticks were found in herds sharing water and pastures resources and with a history of seasonal transhumance. Therefore, common grazing and the seasonal transhumance are likely to support the transmission of the virus. This could have important health and economic impacts, especially regarding transboundary cattle movements.

Control of Dermanyssus gallinae (De Geer 1778) and other mites with volatile organic compounds, a review.

Dermanyssus gallinae (De Geer 1778), commonly named the poultry red mite (PRM), is considered to be the most harmful ectoparasite in poultry farms in Europe. This species feeds on the blood of laying hens, but spends most of its time hidden in cracks and crevices around hen nests. To control PRM populations in poultry houses, chemical pesticides are currently used; however, concern is growing regarding the harmful residues found in eggs and hens, along with the increased resistance of mites against several compounds. Alternatives to synthetic compounds are now being explored, including vaccines, biological control, physical control and semiochemical control based on the chemical ecology of PRM. This review focused on the different volatile organic compounds (VOCs) identified from D. gallinae and other mite species that have been discovered to control them. Pheromones (aggregation pheromone, sex pheromone and alarm pheromone) and kairomones promoting attraction behaviour in D. gallinae and other mite species are presented, while VOCs from essential oils and plant extracts with repellent properties are also explored. Finally, devices using VOCs on PRM in the field are described, with devices that have been tested on other Acari species being mentioned as potential directions for the future control of PRM.

Identification of candidate transmission-blocking antigen genes in Theileria annulata and related vector-borne apicomplexan parasites.

BACKGROUND: Vector-borne apicomplexan parasites are a major cause of mortality and morbidity to humans and livestock globally. The most important disease syndromes caused by these parasites are malaria, babesiosis and theileriosis. Strategies for control often target parasite stages in the mammalian host that cause disease, but this can result in reservoir infections that promote pathogen transmission and generate economic loss. Optimal control strategies should protect against clinical disease, block transmission and be applicable across related genera of parasites. We have used bioinformatics and transcriptomics to screen for transmission-blocking candidate antigens in the tick-borne apicomplexan parasite, Theileria annulata. RESULTS: A number of candidate antigen genes were identified which encoded amino acid domains that are conserved across vector-borne Apicomplexa (Babesia, Plasmodium and Theileria), including the Pfs48/45 6-cys domain and a novel cysteine-rich domain. Expression profiling confirmed that selected candidate genes are expressed by life cycle stages within infected ticks. Additionally, putative B cell epitopes were identified in the T. annulata gene sequences encoding the 6-cys and cysteine rich domains, in a gene encoding a putative papain-family cysteine peptidase, with similarity to the Plasmodium SERA family, and the gene encoding the T. annulata major merozoite/piroplasm surface antigen, Tams1. CONCLUSIONS: Candidate genes were identified that encode proteins with similarity to known transmission blocking candidates in related parasites, while one is a novel candidate conserved across vector-borne apicomplexans and has a potential role in the sexual phase of the life cycle. The results indicate that a 'One Health' approach could be utilised to develop a transmission-blocking strategy effective against vector-borne apicomplexan parasites of animals and humans.

Guidelines for the Detection of Babesia and Theileria Parasites.

The genera Babesia and Theileria (phylum Apicomplexa, order Piroplasmida) are mainly transmitted by Ixodid ticks in which the sexual part of their life cycle followed by sporogony takes place. They include protozoan parasites that infect erythrocytes of a variety of vertebrate hosts, including domestic and wild animals, with some Babesia spp. also infecting humans. Babesia sporozoites transmitted in the tick's saliva during the bloodmeal directly infect erythrocytes, where they asexually multiply to produce pear-shaped merozoites in the process of merogony; whereas a pre-erythrocytic schizogonic life stage in leukocytes is found in Theileria and precedes merogony in the erythrocytes. The wide spectrum of Babesia and Theileria species and their dissimilar characteristics with relation to disease severity, transmission, epidemiology, and drug susceptibility stress the importance of accurate detection of babesiosis and theileriosis and their causative agents. These guidelines review the main methods currently used for the detection of Babesia and Theileria spp. for diagnostic purposes as well as epidemiological studies involving their vertebrate hosts and arthropod vectors. Serological methods were not included once they did not indicate current infection but rather exposure.

Prevalence of Angiostrongylus vasorum in southern Belgium, a coprological and serological survey.

BACKGROUND: Canine angiostrongylosis, a gastropod-borne helminthic infection, is increasingly being described in North America and is now reported in many European countries. In dogs, Angiostrongylus vasorum may cause a wide spectrum of clinical signs. Respiratory distress such as coughing and dyspnoea are the most frequently described manifestations. The aim of the present study was to gain additional information on the distribution, prevalence and risk factors associated with A. vasorum infection in dog from southern Belgium through the combined used of a commercially available in-clinic assay for detection of circulating antigen (Angio Detect™, IDEXX, Westbrook, USA) and coprology in two different canine populations: dogs with clinical signs compatible with angiostrongylosis and asymptomatic dogs or dogs presented for unrelated conditions (control). RESULTS: A total of 979 dogs were enrolled in the study from November 2014 until February 2016. Seven hundred fifty-seven dogs were included in the control group, whereas 222 dogs had clinical signs compatible with angiostrongylosis. Forty-six dogs out of 979 (4.7 %) had A. vasorum circulating antigen. There was a highly significant difference between the two populations (3.6 % (27/747) and 8.6 % (19/222) in control and symptomatic dogs, respectively) (P = 0.00379). First stage larvae (L1) of A. vasorum were found in seven out of 24 serologically positive control dogs and in six out of 17 serologically positive symptomatic dogs. Interestingly, L1 of Crenosoma vulpis were detected by Baermann technique in one control and nine symptomatic dogs, respectively. Out of 17 Angio Detect™ (IDEXX, Westbrook, USA) positive dogs with negative (14) or not performed Baermann test (three), one dog was positive in both in-house ELISAs (Ag and Ab) and one dog was positive for Ag. Statistical analysis was unable to detect any risk factors associated with the direct and/or indirect detection of A. vasorum. CONCLUSIONS: This seroepidemiological study demonstrated for the first time a high seroprevalence in Southern Belgium for A. vasorum. The Angio Detect™ was found to be suitable in this context as the collection, preservation and examination of stools were difficult. Nevertheless, discrepancies were observed between the different available tests. Additional research is clearly needed. Also, coproscopy remains a very useful tool in dogs infected for less than nine weeks and for the identification of other canine lung nematodes such as C. vulpis. This study also demonstrates that asymptomatic dogs may shed A. vasorum L1 in their faeces and therefore contribute to the maintenance of A. vasorum life-cycle.

First report of a fatal autochthonous canine Angiostrongylus vasorum infection in Belgium.

Canine angiostrongylosis is considered as an emergent disease in Europe and Canada. A fatal case of Angiostrongylus vasorum infection is described in a four and a half month old puppy born in Belgium. The dog was presented with marked neurological disorders, body weight loss, a profound weakness and mild respiratory signs. The dog was given antibiotics and mucolytic compounds with very little improvement and consequently was referred to a specialist for additional examinations. As the general condition of the dog was rapidly declining, the animal was euthanized shortly after on its owners' request and a necropsy was carried out. Extensive gross pulmonary lesions were observed and histopathological examination revealed the presence of numerous larvae with morphology compatible with A. vasorum. Larvae were also found in the product of a bronchoalveolar lavage but fecal material was not examined. The presence of A. vasorum circulating serum antigen was demonstrated through ELISA; additionally an A. vasorum specific PCR was performed on brain material and yielded a positive result. This case confirms that the clinical diagnosis of canine angiostrongylosis can be very challenging especially when respiratory signs are absent or very mild such in the present case. This is the first reported case of canine angiostrongylosis in Belgium.

New insight in lymnaeid snails (Mollusca, Gastropoda) as intermediate hosts of Fasciola hepatica (Trematoda, Digenea) in Belgium and Luxembourg.

BACKGROUND: The present study aims to assess the epidemiological role of different lymnaeid snails as intermediate hosts of the liver fluke Fasciola hepatica in Belgium and Luxembourg. METHODS: During summer 2008, 7103 lymnaeid snails were collected from 125 ponds distributed in 5 clusters each including 25 ponds. Each cluster was located in a different biogeographic area of Belgium and Luxembourg. In addition, snails were also collected in sixteen other biotopes considered as temporary wet areas. These snails were identified as Galba truncatula (n = 2474) (the main intermediate host of F. hepatica in Europe) and Radix sp. (n = 4629). Moreover, several biological and non-biological variables were also recorded from the different biotopes. DNA was extracted from each snail collected using Chelex® technique. DNA samples were screened through a multiplex PCR that amplifies lymnaeid internal transcribed spacer 2 gene sequences (500-600 bp) (acting as an internal control) and a 124 bp fragment of repetitive DNA from Fasciola sp. RESULTS: Lymnaeid snails were found in 75 biotopes (53.2%). Thirty individuals of G. truncatula (1.31%) and 7 of Radix sp. (0.16%) were found to be positive for Fasciola sp. The seven positive Radix sp. snails all belonged to the species R. balthica (Linnaeus, 1758). Classification and regression tree analysis were performed in order to better understand links and relative importance of the different recorded factors. One of the best explanatory variables for the presence/absence of the different snail species seems to be the geographic location, whereas for the infection status of the snails no obvious relationship was linked to the presence of cattle. CONCLUSIONS: Epidemiological implications of these findings and particularly the role of R. balthica as an alternative intermediate host in Belgium and Luxembourg were discussed.