Bertrand Losson

Culicoides trapping with Rothamsted suction traps before and during the bluetongue epidemic of 2006 in Belgium

The collection of biting midges was taking place some months before the first bluetongue outbreak in Belgium in August 2006. The Walloon Agricultural Research Centre had been monitoring aphid populations at two sites annually in Belgium (Gembloux and Libramont), using two stationary 12-m Rothamsted suction traps. For the Gembloux trap, collections of insects captured daily from 11 May 2006 onwards were already available at the time of the outbreak. An examination of these samples revealed the presence of Culicoides, some species of which are considered as potential vectors of the bluetongue virus (BTV). The trapping was therefore extended beyond the normal aphid activity period and the Culicoides captured were identified to species level. From 11 May to 31 December 2006, the Gembloux trap caught 664 Culicoides specimens belonging to 19 species comprising known BTV-vectors. The second trap, at Libramont, was reactivated from 12 September to 13 October and caught 97 specimens belonging to nine species, all of which had been found at the Gembloux site. Among the 19 species identified, four were new to Belgian fauna: Culicoides achrayi, C. deltus, C. lupicaris and C. newsteadi. This paper examines the overall phenology and the physiological status of Culicoides in 2006 before and during the bluetongue epidemic. It discusses the potential of the Rothamsted suction trap to monitor Culicoides.

Foci report on indigenous Dermacentor reticulatus populations in Belgium and a preliminary study of associated babesiosis pathogens

The occurrence of autochthonous clinical cases of canine and equine babesiosis in Belgium during the last two decades suggests that the vector of the pathogens responsible for these diseases, Dermacentor reticulatus (Ixodida: Ixodidae), may be present in this country. Consequently, evidence for the presence of this tick species in different locations within Belgium was investigated. Four different locations were monitored by flagging in 2010; these included the locations at which D. reticulatus was previously found on a dog in 2009 and on two red deer in 2007. Two different species of tick were identified, Ixodes ricinus (Ixodida: Ixodidae) and D. reticulatus. A total of 282 D. reticulatus adult ticks (98 males, 184 females) were collected from the four sites. Ticks were found mainly from early March until the end of May and a peak in activity was apparent in March. A Babesia spp. (Piroplasmida: Babesiidae) genus‐specific polymerase chain reaction test based on the amplification of a fragment of the 18S rRNA gene was used to investigate the potential presence of Babesia spp. All DNA extracts isolated from the total tick samples yielded negative results. Additional studies to accurately determine the distribution and vectorial capacity of this important tick species in Belgium are warranted.

A retrospective serological survey on human babesiosis in Belgium

In Europe, most clinical babesiosis cases in humans have been attributed to Babesia divergens and Babesia sp. EU1. Babesia microti infection of humans occurs mainly in the United States; although a case of autochthonous B. microti infection and serological evidence of infection have been reported in Europe. The Indirect Fluorescent Antibody Test was used to screen sera from 199 anonymous Belgian patients with history of tick bite and clinical symptoms compatible with a tick-borne disease. The serological screen detected positive reactivity in 9% (nxa0=xa018), 33.2% (nxa0=xa066), and 39.7% (nxa0=xa079) of the samples against B. microti, B. divergens, and Babesia sp. EU1, respectively. Thus, evidence of contact among three potentially zoonotic species of Babesia and humans has been confirmed in Belgium. Preventive action and development of better diagnostic tools should help in prevention of clinical cases and to clarify the true burden of such infection for individuals and public health.

Detection of Angiostrongylus vasorum by quantitative PCR in bronchoalveolar lavage fluid in Belgian dogs

OBJECTIVESnTo describe Angiostrongylus vasorum infection in a series of clinical cases over a 12-month period, report the use of quantitative PCR on broncho-alveolar lavage fluid and investigate the -possibility of previously undiagnosed angiostrongylosis in a retrospective cohort of coughing and healthy dogs.nnnMETHODSnPulmonary angiostrongylosis was diagnosed based on compatible clinical signs and positive quantitative PCR on broncho-alveolar lavage fluid and/or positive Baermann examination. Quantitative PCR was also performed retrospectively on broncho-alveolar lavage fluid from 65 dogs (55 coughing and 10 healthy dogs), examined between 2008 and 2014; results of Baermann examination were also available in 33 dogs.nnnRESULTSnSeven young adult dogs from Southeastern Belgium with respiratory clinical signs were diagnosed with angiostrongylosis between March 2013 and April 2014. Positive broncho-alveolar lavage fluid quantitative PCR results and positive Baermann examination were obtained in 5/5 and 2/5 dogs, respectively. In the remaining two dogs, only Baermann analysis was performed. Among the retrospective cohorts, only one broncho-alveolar lavage fluid from a coughing dog was quantitative PCR-positive whereas all faecal samples were negative.nnnCLINICAL SIGNIFICANCEnUntil recently, canine angiostrongylosis was not reported in Belgium. It should now be included in the differential diagnosis of coughing Belgian dogs. Identification of affected dogs may be aided by quantitative PCR on broncho-alveolar lavage fluid.

Swimmer's Itch in Belgium: First Recorded Outbreaks, Molecular Identification of the Parasite Species and Intermediate Hosts.

BACKGROUNDnCercarial dermatitis or swimmers itch is a skin condition in humans due to the larval forms of bird schistosomes of some species of the genus Trichobilharzia. The life cycle of these schistosomes requires freshwater snails (intermediate host) and waterfowl (definitive host). Repeated exposures to cercariae can lead to skin sensitization with the induction of pruritic skin lesions.nnnMETHODSnWe describe, in this study, two outbreaks of human cercarial dermatitis at the Eau dHeure Lakes, Belgium. In July and August 2012, a total of, respectively, 78 and 10 people reported a sudden skin rash accompanied by pruritus following recreational activities in the Plate Taille Lake. However, no ocellate furcocercariae were detected following light exposure of the snails collected between September 2012 and September 2013 (nu2009=u2009402). No outbreaks were recorded in 2013 and 2014. In August 2015, about 30 new cases were recorded. Snails were collected (nu2009=u2009270) in different locations around the lake. PCR was used to identify accurately the intermediate hosts and the parasite species involved.nnnRESULTSnAfter light exposure, seven Radix spp. (2.6%) shed ocellate furcocercariae. Molecular identification based on the rDNA internal transcribed spacer 2 sequence ascribed the infected snails to R. balthica (u2009=u2009R. peregrau2009=u2009R. ovata) (6/7) and R. auricularia (1/7). Based on the amplification of the D2 domain of the 28S rDNA, the cercariae were shown to belong to two different haplotypes of Trichobilharzia franki.nnnCONCLUSIONSnThis is the first record in Belgium of T. franki and associated skin condition.

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

BackgroundCanine 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).ResultsA 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 (4.7xa0%) had A. vasorum circulating antigen. There was a highly significant difference between the two populations (3.6xa0% (27/747) and 8.6xa0% (19/222) in control and symptomatic dogs, respectively) (Pu2009=u20090.00379). 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.ConclusionsThis 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 evidence of Besnoitia bennetti infection (Protozoa: Apicomplexa) in donkeys (Equus asinus) in Belgium

BackgroundBesnoitiosis is caused by different species of intracellular protozoan parasites belonging to the family Sarcocystidae and affecting multiple host species worldwide. Including B. besnoiti, ten species are described infecting animals. Among ungulates, Besnoitia bennetti infects horses, donkeys and zebras and was described in Africa and in the USA where donkey besnoitiosis is considered as an emerging disease.Case presentationA two-year-old male donkey was purchased in May 2016 in poor body condition (cachexia, alopetic areas and pruritus mainly on neck and head) by the present owner in Le Roeulx (Belgium) from a milk producing donkey farm in Frasnes-lez-Buissenal (Belgium). Shortly after its purchase and shearing, the donkey presented with crusts, hyperkeratosis (both flanks and neck) anorexia and cachexia. A treatment with phoxim was given with no improvement. A cutaneous biopsy of hyperkeratotic skin was performed in July. It showed a perivascular eosinophilic infiltrate with a large thick walled cyst located in the dermis containing numerous bradyzoites. This was highly suggestive of besnoitiosis. Several skin biopsy samples were obtained for qPCR analysis and confirmed the presence of Besnoitia spp. DNA. Further laboratory diagnosis tests were performed (western blot and rDNA sequencing) confirming Besnoitia bennetti aetiology for the male. For the female, the punch-biopsy, haematology and qPCR were negatives but the western blot showed the presence of antibodies directed to Besnoitia spp. Further clinical examination performed in August highlighted scleral pinhead sized cysts (pearl) in the right eye and between nares. Another ten-year-old female donkey purchased in France and sharing the same accommodation showed a good clinical condition, but a thorough clinical examination showed the presence of numerous cysts on the inner face of upper labial mucosa. A daily treatment based on sulfamethaxzole and trimethoprim (Emdotrim 60% Mix®, 30 mg/kg) was given orally and some improvement was noticed.ConclusionThis is the first evidence of Besnoitia bennetti infection (Protozoa: Apicomplexa) in donkeys (Equus asinus) in Belgium.

Alveolar echinococcosis in southern Belgium: retrospective experience of a tertiary center.

Dear Editor, Alveolar Echinococcosis (EA) is a zoonosis due to the larval stage of the fox tapeworm Echinococcus multilocuris. Humans are dead-end hosts and are exposed through sylvatic (fox) or domestic (cat and dog) cycles. Infection is acquired through the fecal-oral route. The metacestodes of E. multilocularis proliferate in the liver, inducing a Btumorlike^ lesion that can invade the neighboring organs or spread away from the primary lesion [1]. Until recently, Belgium was considered as a low-risk country for AE. However, in 2008, Hanosset et al. demonstrated by necropsies of red foxes (Vulpes vulpes), a prevalence of AE at up to 60% in some parts of Wallonia, the Southern part of Belgium [2]. The first indigenous Belgian human AE case was diagnosed in 1999 at the Centre Hospitalier Universitaire (CHU) of Liege, a tertiary university hospital in Wallonia [3]. Since this first case, other patients have been diagnosed with EA and managed by the different departments of the CHU Liege [4]. The aim of this study was to evaluate the overall experience and results of the different teams of the CHU Liege with AE and to better determine the number of indigenous AE cases to provide this information to authorities in charge of public health. After University Hospital Ethical committee approval, the authors retrospectively collecteddata from the laboratoryof clinical microbiology (for Echinococcus serologies and PCR), the hospital pharmacy in charge of supplying albendazole, and by searching through patient files from themedico-economic informationservice. Informationwascollectedfrom1999toFebruary 2018. Belgian regulations do not require patient informed consent for a purely retrospective review of medical files. Between 1999 and February 2018, a total of 22 human indigenous AE cases were recorded and their medical files were studied. In all cases, the diagnosis was established based on Echinococcus sp. serology (inhibition of hemagglutination (Fumouze, France), ELISA specific for E. granulosus (Rbiopharm, Germany) and E. multilocularis (Bordier, Suisse) respectively and Western Blot), clinical imaging, histopathology and in some cases an E. multilocularis specific PCR assay on tissue [5]. According to the criteria of Brunetti et al. [1], 11 possible and 11 confirmed cases were diagnosed. The mean age of the patients at the time of diagnosis was 69 years (ranges: 34–85 years). Sixty-four percent of the patients were male. Some degree of immunosuppression could be identified in 36% of cases (solid or hematologic cancers, chronic inflammatory disease, diabetes, and chronic alcoholism). At least one of the risk factors described by Conraths et al. [6] (owning a dog and/or a cat, living in a rural zone, working as farmer, or forestry worker) was identified in all patients but one (data are missing). Patients lived in rural * Audrey Cambier au.cambier@gmail.com

Hepatic alveolar echinococcosis.

Olivier Detry, Nicolas Meurisse, Jean Delwaide, Jean-Baptiste Giot, Philippe Leonard, Bertrand Losson, Marie-Pierre Hayette, Noella Bletard, Paul Meunier and Pierre Honor e Department of Abdominal Surgery and Transplantation, CHU Liege, University of Liege (CHU-ULg), Liege, Belgium; Multidisciplinary Unit for Echinococcosis Management and Research of the University of Liege (Echino-Liege), University of Liege, Liege, Belgium; Department of Hepatogastroenterology, CHU Liege, University of Liege (CHU-ULg), Liege, Belgium; Department of Infectious diseases, CHU Liege, University of Liege (CHU-ULg), Liege, Belgium; Laboratory of Parasitology and Pathology of Parasitic Diseases, Faculty of Veterinary Medicine, University of Liege (ULg), Liege, Belgium; Department of Microbiology, CHU Liege, University of Liege (CHU-ULg), Liege, Belgium; Department of Pathology, CHU Liege, University of Liege (CHU-ULg), Liege, Belgium; Department of Radiology, CHU Liege, University of Liege (CHU-ULg), Liege, Belgium

Gastrointestinal parasites in captive and free-ranging Cebus albifrons in the Western Amazon, Ecuador

Currently, there is a lack of surveys that report the occurrence of gastrointestinal parasites in the white-headed capuchin monkey (Cebus albifrons). We therefore assessed the presence and richness (= number of different parasite genera) of parasites in C. albifrons in wildlife refuges (n = 11) and in a free-ranging group near a human village (n = 15) in the Ecuadorian Amazon. In the 78 samples collected (median of 3 samples per animal), we identified a total of 6 genera of gastrointestinal parasites, representing protozoa, nematodes, acanthocephalans and cestodes. We observed a high prevalence (84%) across the 26 individuals, with the most prevalent parasite being Strongyloides sp. (76.9%), followed by Hymenolepis sp. (38.5%) and Prosthenorchis elegans (11.5%). We found Entamoeba histolytica/dispar/moskovskii/nuttalli and Capillaria sp. in only a minority of the animals (3.8%). In addition, we observed unidentified strongyles in approximately one-third of the animals (34.6%). We found a total of 6 parasite genera for the adult age group, which showed higher parasite richness than the subadult age group (5) and the juvenile age group (3). Faecal egg/cyst counts were not significantly different between captive and free-ranging individuals or between sexes or age groups. The free-ranging group had a higher prevalence than the captive group; however, this difference was not significant. The only genus common to captive and free-ranging individuals was Strongyloides sp. The high prevalence of gastrointestinal parasites and the presence of Strongyloides in both populations support results from previous studies in Cebus species. This high prevalence could be related to the high degree of humidity in the region. For the free-ranging group, additional studies are required to gain insights into the differences in parasite prevalence and intensity between age and sex groups. Additionally, our study demonstrated that a serial sampling of each individual increases the test sensitivity.