Mind the gaps in research on the control of gastrointestinal nematodes of farmed ruminants and pigs.

Gastrointestinal (GI) nematode control has an important role to play in increasing livestock production from a limited natural resource base and to improve animal health and welfare. In this synthetic review, we identify key research priorities for GI nematode control in farmed ruminants and pigs, to support the development of roadmaps and strategic research agendas by governments, industry and policymakers. These priorities were derived from the DISCONTOOLS gap analysis for nematodes and follow-up discussions within the recently formed Livestock Helminth Research Alliance (LiHRA). In the face of ongoing spread of anthelmintic resistance (AR), we are increasingly faced with a failure of existing control methods against GI nematodes. Effective vaccines against GI nematodes are generally not available, and anthelmintic treatment will therefore remain a cornerstone for their effective control. At the same time, consumers and producers are increasingly concerned with environmental issues associated with chemical parasite control. To address current challenges in GI nematode control, it is crucial to deepen our insights into diverse aspects of epidemiology, AR, host immune mechanisms and the socio-psychological aspects of nematode control. This will enhance the development, and subsequent uptake, of the new diagnostics, vaccines, pharma-/nutraceuticals, control methods and decision support tools required to respond to the spread of AR and the shifting epidemiology of GI nematodes in response to climatic, land-use and farm husbandry changes. More emphasis needs to be placed on the upfront evaluation of the economic value of these innovations as well as the socio-psychological aspects to prioritize research and facilitate uptake of innovations in practice. Finally, targeted regulatory guidance is needed to create an innovation-supportive environment for industries and to accelerate the access to market of new control tools.

Species composition of larvae cultured after anthelmintic treatment indicates reduced moxidectin susceptibility of immature Cylicocyclus species in horses.

For the control of cyathostomins in horses, the macrocyclic lactones (MLs), moxidectin (MOX) and ivermectin (IVM) are the most commonly used anthelmintics. However, reduced activity, observed as shortening of the egg reappearance period (ERP) has been described. Shortening of the ERP may be caused by a decreased susceptibility of immature worms for MLs. Alternatively, immature worms may develop faster into egg producing adults as a result of repeated ML treatments. The species composition of the larval cultures obtained shortly after ML and pyrantel (PYR) treatment can confirm the hypothesis of decreased ML susceptibility, as this is often class-specific, whereas faster development would also occur after treatment with anthelmintics with a different mode of action. From 3 farms with a known history of shortened ERP, 8 horses per farm were selected and divided into 2 groups. The MOX-PYR-MOX group was treated twice with MOX (day 0 and 126) and once with PYR (day 84) and the IVM-PYR-IVM group was treated twice with IVM (day 0 and 98) and once with PYR (day 56). Cultured infective larvae (L3s) were counted and differentiated with the reverse line blot on pooled samples. Per cyathostomin species, the number of larvae per gram was calculated. The efficacy of all ML treatments was 100% and a shortened ERP was found on all 3 farms. The species composition of the larval cultures after ML treatment did not differ significantly from that after PYR treatment in the IVM-PYR-IVM group, but it did differ in the MOX-PYR-MOX group. The larval cultures obtained after MOX treatment consisted mostly of Cylicocyclus nassatus, while after PYR treatment Cylicostephanus longibursatus was the most abundant species. In the cultures from 42days after MOX treatment 6 cyathostomin species from 3 genera were found on the farm with the lowest activity (farm 1), while on the farm with the highest activity (farm 3) only 3 species from one genus were found in the same number of examined L3s. The high numbers of L3s of Cylicocyclus species 42days after MOX treatment and the low numbers 42days after PYR treatment can be explained by reduced susceptibility of the immature worms to MOX, but not by a faster development. In conclusion, shortening of the ERP following MOX treatment is most likely a process in which an increasing number of immature worms from an increasing number of species is becoming less susceptible to the active compound.

Semi-quantitative differentiation of cyathostomin larval cultures by reverse line blot.

Cyathostomins are the most prevalent horse nematodes worldwide and over 50 species are described. The eggs and the infective larvae (L3) can easily be obtained or cultured from infected horses, but cannot be differentiated morphologically at species level. A reverse line blot (RLB) method based on the hybridization of a PCR fragment with a species specific probe, has previously been developed for the differentiation of individual eggs and/or L3s, but is too labor intensive for large scale studies. In the present study a RLB method on multiple pooled L3s for the semi-quantitative differentiation of cyathostomin larval cultures was developed and validated. First, the probability of the presence of a certain species within a pool was calculated as function of the frequency and the number of L3s within a pool. Ten L3s per pool were found to be optimal. Next, the probability, the chance of occurrence was calculated when 4 pools per culture were used. The probability distributions for 0, 1, 2, 3 or 4 positive pools were transformed into the corresponding median frequency of the cumulative probability: 0.014, 0.04, 0.08, 0.16 and 0.59, respectively. Based on these calculated probabilities, RLB on 10 L3s per pool and 4 pools per sample was validated by estimating the cross-hybridization, precision and accuracy in 3 groups of horses. First, absence of cross-hybridization was confirmed by differentiation of the same L3s (160 L3s from the 4 horses from group 1) in the RLB on individual as well as on pooled L3s. Cross-hybridization was excluded for 9 of the most common cyathostomins. Next, the precision and accuracy were determined by the differentiation of 10 replicates of 3 cultures from 3 horses from group 2 (1200 L3s). The coefficient of variation (CV) was between 0 and 0.90 and the accuracy was between 0.42 and 1.73. A Monte Carlo simulation based on the observed scores and associated probability distributions gave similar results as the use of a fixed median frequency. The LPGs obtained from 276 larval culture counts from a larger cohort (23 horses, group 3) were not significantly different from the LPGs obtained from summation of the LPG per species found by RLB on pooled L3s. The RLB on pooled L3s was found therefore an useful semi-quantitative method for the differentiation of the most common cyathostomin L3, with a workload of approximately one tenth of that of the RLB on individual L3s.

Resistance of gastrointestinal nematodes to the most commonly used anthelmintics in sheep, cattle and horses in Spain.

The objective of this study was to evaluate the status of anthelmintic resistance (AR) in ruminants and horses in Spain. The efficacy of commonly used macrocyclic lactones (MLs) - ivermectin (IVM) and moxidectin (MOX) - was measured in sheep, cattle and horses. In addition, albendazole (ABZ) and levamisole (LEV) were evaluated in sheep and oxibendazole (OXI) and pyrantel (PYR) in horses. Efficacy was evaluated based on the difference between the arithmetic mean pre- and post-treatment faecal egg count (in cattle and horses), or compared to an untreated control group (in sheep). AR was present when the percentage reduction in egg count was <95% and the lower 95% confidence interval (CI) was <90%; if only one of these two criteria was met, the finding was recorded as suspected AR (SAR). In horses, AR-PYR and OXI was considered when the percentage reduction in egg count was ≤ 90% and the lower 95% CI was ≤ 80%. For each animal species, at least 10 study sites were selected. AR to at least one of the drugs was detected in all 10 sheep flocks; the main parasite identified after treatment was Teladorsagia circumcincta. Moreover, in 5 flocks multidrug resistance was identified, on 4 farms to drugs from different families, on one farm to both MOX and IVM and on another farm to all drugs tested. In cattle, the efficacy of both MOX and IVM was 100% on 4 and 3 farms, respectively, and therefore 60% of these farms were considered to have AR or SAR to both MLs. The most frequent parasite identified after treatment was Trichostrongylus spp., although Ostertagia ostertagi was also identified after treatment on one farm. In contrast to ruminants, the 4 drugs evaluated in horses were highly efficacious against strongyles, with efficacies for the MLs and OXI between 95 and 100% and between 94 and 100% for PYR, although 3 herds were SAR against PYR. In conclusion, AR to at least one of the commonly used drugs was identified on all sheep flocks investigated in the northwest of Spain. The occurrence of AR to MLs in cattle was higher than expected but consistent with what was observed in sheep. In horses, all currently used drugs were confirmed as effective against strongyles.

The occurrence and genetic characterization of Cryptosporidium and Giardia species in foals in Belgium, The Netherlands, Germany and Greece.

Faecal samples were collected from foals between the age of 1 week and 6 months in Belgium, The Netherlands, Germany and Greece. A quantitative direct immunofluorescence assay based on the commercial MERIFLUOR Cryptosporidium/Giardia kit was performed to evaluate the presence of (oo) cysts. Parasite positive samples were genotyped, based on the 18S ribosomal DNA gene and the heat shock protein (HSP70) gene for Cryptosporidium and on the ß-giardin gene and the triose phosphate isomerase (TPI) gene for Giardia. In total, 134 foals from Belgium, 44 foals from The Netherlands, 30 foals from Germany and 190 foals from Greece were examined. No Cryptosporidium oocysts were identified in faecal samples from foals in Germany and The Netherlands. In Belgium and Greece, 4.5% and 1.1% of the foals examined were Cryptosporidium positive, respectively, all with a low oocyst excretion ranging from 100 to 2450 oocysts per gram of faeces. For Giardia, 14.2%, 11.4%, 10.0% and 11.6% of the foals in Belgium, The Netherlands, Germany and Greece, respectively, were found to excrete cysts, with a range of 50 up to 4,000,000 cysts per gram of faeces. Younger animals secreted significantly more Giardia cysts than older horses (p<0.05), but no significant correlation between Giardia infection and diarrhoea was observed. Most Giardia positive samples belonged to assemblage AI and/or BIV, but also assemblage E was detected in two samples. Together with the identification of Cryptosporidium horse genotype, this suggests only a low risk for zoonotic transmission.

Cylicocyclus species predominate during shortened egg reappearance period in horses after treatment with ivermectin and moxidectin.

The normal time interval between treatment of horses with a macrocyclic lactone (ML) and reappearance of strongyle eggs in the feces, or 'Egg Reappearance Period (ERP)', is at its shortest 8 weeks for ivermectin (IVM) and 12 weeks for moxidectin (MOX). Nowadays, it is not uncommon to find shorter ERPs, potentially indicating the beginning of the development of drug resistance. Whether all cyathostomin species contribute equally to a shortened ERP is not known. In the present study a Reverse Line Blot (RLB) on individual infective larvae was used to compare species composition before and after ML-treatment in horses with either a normal or a shortened ERP. Species composition was determined for a total of 21 horses on 4 farms and was based on differentiation of approximately 40 larvae per horse per sampling day. After IVM treatment the shortest ERP was found to be 28-42 days. After MOX treatment the shortest ERP was found to be 42-56 days. The RLB showed that early egg shedding was dominated by species of the genus Cylicocyclus (Cyc.) (Cyc. ashworthi, Cyc. insigne, Cyc. leptostomum and Cyc. nassatus). Before treatment, species composition was generally more diverse with species from several other genera also present. Interestingly, on the farm with the overall shortest ERP and where all horses showed a shortened ERP, more species/genera were present during early egg excretion than on other farms. Results suggest that cyathostomin species within the genus Cylicocyclus contribute to a shortened ERP more than species from other genera. However, if shortening of ERP overall becomes more pronounced and widespread within a farm, species from other genera will also be present during early egg shedding.

World Association for the Advancement of Veterinary Parasitology (WAAVP): Guideline for the evaluation of drug efficacy against non-coccidial gastrointestinal protozoa in livestock and companion animals.

The current guideline was written to aid in the design, implementation and interpretation of studies for the assessment of drug efficacy against non-coccidial gastrointestinal protozoan parasites, with Giardia spp. as the leading example. The information provided in this guideline deals with aspects of how to conduct controlled studies using experimental infection models (dose determination and dose confirmation) and efficacy studies in commercial facilities (field effectiveness studies). Furthermore, the selection of suitable animals, housing, infection procedure, choice of diagnostic technique and data analysis are discussed. This guideline is intended to assist investigators in conducting specific studies, to provide specific information for registration authorities involved in the decision-making process, to assist in the approval and registration of new drugs and to facilitate the worldwide adoption of uniform procedures. The primary parameter for drug efficacy is the reduction in either parasite excretion or parasite counts and a minimum efficacy of 90% is required against non-coccidial gastrointestinal protozoa. A supporting efficacy parameter is a significant difference in the proportion of infected animals between treated and non-treated groups. Persistent efficacy is considered as an additional claim to therapeutic efficacy.

Production benefit of treatment with a dual active oral formulation of derquantel-abamectin in slaughter lambs.

Gastrointestinal nematodes have a negative impact on the productivity of grazing sheep. Control of these infections has relied for decades on the use of anthelminics, leading to anthelmintic resistance (AR) worldwide, including the UK. The use of combination anthelmintics has been advocated as one of the tools in managing AR and decreasing the economic losses associated with AR. In this study, the benefit of a new combination (derquantel plus abamectin; DQL+ABA: Startect(®) Dual Active Oral Solution for Sheep, Zoetis, at 1 mL/5 kg bodyweight) was evaluated on a farm with suspected AR to benzimidazoles, by incorporating it into the existing treatment regime. In total 100 lambs were selected and allocated to one of two treatment groups, which were treated either with DQL+ABA or fenbendazole/levamisole at monthly intervals. All lambs were co-grazing the same pasture. Weight gain and carcass quality was evaluated, as well as treatment efficacy. The efficacy of DQL+ABA was high throughout the grazing season, whereas the efficacy of the fenbendazole/levamisole treatment regime was below the threshold for anthelmintic efficacy. The high efficacy of a monthly treatment with DQL+ABA resulted in a significantly higher average daily weight gain and shorter time to reach slaughter.

Evaluation of cellular and humoral systemic immune response against Giardia duodenalis infection in cattle.

Giardia duodenalis causes diarrhoea in humans and a wide range of mammals, including cattle. In cattle, the infection often has a chronic character. Infected calves may excrete cysts for several months, suggesting that Giardia is able to suppress and evade the immune response. In this study six calves were infected with G. duodenalis assemblage A and E and housed in an environment that allowed reinfection. Cyst excretion was monitored twice a week and blood was collected every 2 weeks, until decreasing cyst counts indicated the development of protective immunity. The kinetics of the circulating memory cells and serum antibodies were followed up throughout this period. Cyst excretion started 1 week post-infection and remained high until week 14. Low cyst counts from week 15 p.i. onwards indicated that the calves had developed immunity. From week 5 p.i. significant proliferation of CD4(+) αß T-cells was observed after in vitro stimulation with G. duodenalis antigen. Characterisation of the proliferating CD4(+) T-cells using real time qPCR showed that at the peak of antigen driven PBMC proliferation the majority of cells were CD4(+) T-cells expressing IL-17 and to a lesser extent FoxP3. The cell proliferation was strongly reduced after plastic adhesion of the PBMC, suggesting a role for antigen-presenting cells. Failure to restore proliferation of depleted PBMC with Giardia-stimulated monocyte-derived dendritic cells (MoDC) and unchanged proliferation after depletion of CD21(+) B-cells showed that other antigen-presenting cells than MoDC and B-cells were important for T-cell proliferation. Analysis of the antibody response indicated that serum IgG1 and IgA levels against G. duodenalis assemblage A and E increased from week 11 post-infection. From the start of the antibody response, all trophozoites stained positive in an immunofluorescence assay with serum antibodies, indicating that a broad repertoire of antibodies was produced against all variant-specific surface proteins. Further research is necessary to determine which effector T-cell subset produces IL-17 and which cells play a role in antigen presentation.

Giardia duodenalis stimulates partial maturation of bovine dendritic cells associated with altered cytokine secretion and induction of T-cell proliferation.

Giardia duodenalis is an important intestinal parasite in animals and humans. The role of dendritic cells (DC) in the initiation of the immune response against G. duodenalis is poorly documented. The aim of this study was to test the hypothesis that G. duodenalis interferes with bovine DC function. Therefore, the effect of trophozoites and excretion/secretion products on bovine monocyte-derived dendritic cells (MoDC) was investigated. We assessed MoDC maturation and cytokine production of G. duodenalis-stimulated MoDC and the ability of these MoDC to take up antigen and induce lymphocyte proliferation. Little or no upregulation of maturation markers CD40 and CD80 was measured, but MHCII expression was increased after stimulation with low parasite concentrations. A dose-dependent decrease in ovalbumin uptake was observed in G. duodenalis-stimulated MoDC. In addition, stimulated MoDC induced proliferation of CD3(-) , γδ-T-cells and TCRαß(+) CD4(+) and CD8(+) T-cells. Increased transcription of TGF-ß was shown in CD4(+) T cells, and increased TNF-α, TGF-ß, IL-10 and IL-4 were seen in γδ-T-cells. We found no evidence that G. duodenalis has a regulatory or inhibitory effect on bovine MoDC. MoDC stimulated with G. duodenalis are functionally active and able to induce proliferation of T cells that produce both pro- and anti-inflammatory cytokines.

A multicentre prevalence study in Europe on Giardia duodenalis in calves, with molecular identification and risk factor analysis.

The present study aimed to obtain data on the occurrence of Giardia duodenalis in calves in four major cattle rearing countries in Europe (Germany, UK, France and Italy), along with genotyping data and risk factors associated with these infections. A total of 2072 calves were sampled on 207 farms. The majority of the animals were Holstein dairy or mixed Holstein calves (n=1565 or 75.5%), and were female (n=1640 or 79.1%). The average age was 7.8 weeks (SD=4.1; median=7; range=2-16 weeks). All fecal samples were tested using a commercially available monoclonal antibody-based ELISA. The overall apparent prevalence of G. duodenalis for the four countries was 45.4% (n=942/2072) and the overall farm prevalence was 89.9% (186/207), with differences in both animal and farm prevalence between the four countries. The prevalence was significantly higher in animals up to 8 weeks (OR=1.88; P<0.001) compared to older calves, and several management factors including contact with the Dam, Frequency of cleaning of the Maternity Pens, and Disinfection of the Calf Housing were found to be associated with infection. Positive samples were withheld for genotyping using the ß-giardin and triose phosphate isomerase gene: G. duodenalis assemblage E was most prevalent, although 43% of the isolates were typed as assemblage A, with differences in between countries. Furthermore, 32% of the examined samples was found to be a mixed assemblage A and E infection, which is consistent with previous reports. The results of the present study confirm previous findings in other European countries that G. duodenalis infections are common in calves. The infection especially occurs in animals younger than 2 months, and the proportion of positive animals gradually decreased with increasing age.

Doramectin resistance in Haemonchus contortus on an alpaca farm in Belgium.

Parasitism by gastrointestinal nematodes is a health concern in New World Camelids (NWC) worldwide, and anthelmintic treatment is often needed for parasite control. Although anthelmintic resistance has been reported in ruminants worldwide, data in NWC are only scarce. In the present study, a case of suspected doramectin resistance in alpacas was examined. A field efficacy study was conducted for the evaluation of two different dosages of doramectin using a faecal egg count reduction test. A group of 8 alpacas was treated with a subcutaneous injection of doramectin at 0.2mg/kg bodyweight. Individual faecal samples were collected before treatment and 7 days after treatment. The faecal egg counts indicated a treatment efficacy of only 68%. To determine whether the treatment failure was caused by true anthelmintic resistance or suboptimal dosage in this animal species, a group of 4 alpacas was subsequently treated at 0.3mg/kg bodyweight. Faecal egg counts 7 days post treatment were reduced by only 41%, indicating that the treatment failure was more likely to be caused by the presence of resistant parasites on this farm. Coprocultures of faecal samples collected after treatment indicated the presence of 98.5% Haemonchus contortus and a small percentage of Cooperia oncophora (<1.5%). A controlled efficacy trial in sheep, for which the optimal dosage of doramectin is known, was conducted to ensure that this truly was a case of resistant parasites. Infective larvae collected from the faeces of these alpacas were used to infect eight nematode-free lambs. These lambs were assigned to one of two groups based on faecal egg counts post infection. One group was treated with doramectin injectable at 0.2mg/kg bodyweight, the other group served as a non treated control group. Pharmacokinetics indicated that the doramectin treatment was adequate, yet an efficacy of only 16% was determined on day 7 after treatment. Identification of the larvae after treatment revealed 100% H. contortus. On day 7 after treatment, H. contortus worm counts were only reduced by 8% in the treated lambs. The results of the present study report for the first time a case of doramectin resistance in alpacas, mainly in H. contortus.

Giardia duodenalis cyst survival in cattle slurry.

Giardia duodenalis is a protozoan parasite known to infect animals and humans. Zoonotic transmission of G. duodenalis can occur by the consumption of drinking water produced from surface water that is contaminated by runoff from manure-laden fields or pastures. Although it was previously reported that storing solid cattle manure decreases G. duodenalis cyst viability, no data are available on cyst survival in slurry waste from cattle. In this study the number and the viability of G. duodenalis cysts was determined in cattle slurry for up to 90 days. G. duodenalis cysts were counted in 30 slurry samples with a quantitative direct immunofluorescence assay. The geometric mean number of cysts was reduced by 77% after 90 days (P<0.0014), although there was substantial variability between samples. A fluorogenic dye staining using 4',6'-di-amino-2-phenylindole and propidium iodide showed a decreased viability from 45 days onwards, and after 90 days incubation, only 3% of the cysts were viable. Gerbils and lambs were artificially infected with 50 day-old and 90 day-old cysts and faecal excretion of G. duodenalis was monitored between 5 and 7 days after infection. Seven days after infection the gerbils were euthanized for Giardia trophozoite counts. Although one cyst was found in the faeces of one of the gerbils after infection with 50 day-old cysts, no trophozoites were recovered from the intestines of any gerbil (n=8). Experimental infection of lambs with 10(5)50 day-old and 90 day-old slurry cysts caused low cyst excretion in one out of two and one out of three lambs, respectively. Together, these data show that storage of cattle slurry for 90 days greatly reduces the number and viability of G. duodenalis cysts.

Mixed Giardia duodenalis assemblage A, B, C and E infections in pet chinchillas (Chinchilla lanigera) in Flanders (Belgium).

Worldwide there is an increasing trend to keep exotic animals as pets. In contrast to domestic animals, few studies have addressed the importance of infectious diseases of these exotic animals harbor. Chinchillas are host to Giardia, but prevalence studies are scarce. Moreover, little is known about their role as a zoonotic reservoir for Giardia. Therefore, the objective of the present study was (1) to study the occurrence of Giardia in pet chinchillas, (2) to identify risk factors and (3) to determine the role of these animals as potential zoonotic reservoir. To this end, pet chinchillas (Chinchilla lanigera) from both pet owners and breeders in Flanders (Belgium) were screened for the presence of Giardia spp. using a sedimentation flotation technique. A questionnaire was distributed among the pet owners to identify putative risk factors. A subset of the Giardiaisolates was characterized by direct sequencing of the ß-giardin gene. In addition, assemblage specific PCRs targeting the triose phosphate isomerase (tpi) gene were performed. Of the 80 chinchillas screened, 53 (66.3%) excreted cysts of Giardia. Youngsters and animals participating in shows were significantly more at risk for infection. A total of 22 isolates were characterized. Direct sequencing of the ß-giardin gene solely revealed the presence of G. duodenalis assemblage B. The assemblage specific PCRs confirmed these findings, but also revealed the presence of assemblage A (11 samples), C (15 samples) and E (2 samples). This study indicates that multiple Giardia spp. are highly prevalent in pet chinchillas and that these animals are a potential reservoir for zoonotic transmission. In addition, the results highlight the benefit of using an assemblage specific PCR in molecular studies as mixed infections are likely to be missed using conventional PCR approaches.

Multilocus genotyping of Cryptosporidium and Giardia in non-outbreak related cases of diarrhoea in human patients in Belgium.

Stool samples from Belgian patients suffering from abdominal pain and/or diarrhoea were examined for Cryptosporidium and Giardia. Cryptosporidium-positive samples were genotyped using the 70 kDa heat shock protein and the 60 kDa glycoprotein (GP60) genes: C. hominis was identified in 54.2% and C. parvum in 45.8% of the samples. Sequencing at the GP60 locus indicated that subgenotype IbA10G2 of C. hominis and subgenotype IIaA15G2R1 of C. parvum were the most prevalent, although several other subgenotypes were identified. For Giardia, sequencing at the beta-giardin, triose phosphate isomerase (TPI) and glutamate dehydrogenase (GDH) genes revealed assemblage B as the most prevalent (74.4%) in human patients. A high degree of heterogeneity was found, especially on the beta-giardin gene, and to a lesser extent on the GDH gene. Furthermore, using a novel species-specific PCR based on the TPI gene, mixed infections with both assemblage A and B were detected in a large number (32.4%) of human patients, which might have important epidemiological implications.

Giardia and other intestinal parasites in different dog populations in Northern Belgium.

The objectives of this study were to obtain data on the prevalence of intestinal parasites in different dog populations in northern Belgium, to estimate the zoonotic risk associated with these infections and to identify potential risk factors. Between 2004 and 2007 a total of 1159 faecal samples were collected from 451 household dogs, 357 dogs from breeding kennels and 351 dogs with gastrointestinal disorders. The samples from dogs with gastrointestinal disorders were sent to the diagnostic Laboratory for Parasitology at Ghent University by veterinary practitioners. In household dogs the prevalence of intestinal parasites was relatively low. Giardia was the most commonly found parasite (9.3%, CI 5.5-13.1), followed by Toxocara canis (4.4%, CI 2.7-6.8). Much higher infection rates were observed in kennel dogs, especially for Giardia spp. (43.9%, CI 37.8-50.0); T. canis (26.3%, CI 21.8-31.2) and Cystoisospora spp. (26.3%, CI 21.8-31.2). Also in dogs with gastrointestinal problems, Giardia spp. (18.1%, CI 13.1-23.1), Cystoisospora spp. (8.8%, CI 6.1-12.3) and T. canis (7.4%, CI 4.9-10.7) were the most frequently detected parasites. In all dog populations pups were more frequently infected with Cystoisospora (P<0.0001 to P<0.05), Giardia (P<0.001 to P<0.05), and T. canis (P<0.0001 to P<0.001) than adult dogs, except for T. canis in household dogs, where this correlation was not significant. A significant association of anthelmintic treatment with T. canis infections was only observed within the household population. Household dogs with a higher number of treatments per year were more frequently infected with T. canis (P<0.05). There was a significant difference between the different breeding kennels for the occurrence of Cystoisospora, Giardia and T. canis (P<0.0001) and large kennels harboured relatively more infected animals than smaller breeding facilities (P<0.05). However, this was not significant for Giardia spp. Breed and gender did not affect the risk of an infection in any of the study populations. Toxocara and Giardia present a zoonotic risk, especially in household dogs, where the majority of Giardia positive samples (80%) belonged to the zoonotic assemblage A. In kennel dogs and clinically affected dogs the host-specific Giardia assemblages C and D were most prevalent (94% and 80%, respectively).

A Bayesian evaluation of three diagnostic assays for the detection of Giardia duodenalis in symptomatic and asymptomatic dogs.

A Bayesian approach was used to evaluate three commonly used diagnostic assays for the detection of Giardia duodenalis in dogs: microscopical examination (ME), a commercial immunofluorescence assay (IFA: MerifluorGiardia test) and a commercial immunochromatographic assay (SNAP: Idexx SNAPGiardia test). These assays were evaluated for use in two different settings: in a cross-sectional epidemiological survey in household dogs and in a clinical survey, both conducted in the northern part of Belgium. A total of 272 faecal samples from household dogs and 141 faecal samples from clinically affected dogs were examined using these three diagnostic assays. The Bayesian analysis indicated that all tests were highly specific (specificity above 90%), and that the IFA is more sensitive than SNAP and ME, both in an epidemiological and in a clinical setting. For all three tests, the estimated sensitivity values were higher in the clinical compared to the epidemiological survey, whereas the specificity values were comparable in both studies. The results of the present study indicate that IFA is a highly specific and sensitive technique for the detection of G. duodenalis cysts, both for use in an epidemiological or clinical survey. The SNAP is a specific and fairly sensitive technique for the diagnosis of Giardia in clinically affected dogs. Overall, the ME was found to be a specific diagnostic technique, although lacking sensitivity.

Parasitic infections in dairy cattle around Hanoi, northern Vietnam.

In northern Vietnam, dairy cattle are mainly managed in small-scale farms, where animals are kept confined and feeding occurs by cut and carry methods. In the present study the occurrence of parasitic infections was examined in five provinces around Hanoi. A total of 201 farms were visited, and 334 stool and 239 blood samples were collected from calves younger than 3 months, animals between 3 and 24 months and adult cows. Furthermore, 254 milk samples were collected from lactating animals. Coproscopical examination indicated a high prevalence of nematode eggs (Cooperia spp., Haemonchus and Oesophagostomum spp.) in animals (n=176) between 3 and 24 months (66%) and in adult cows (n=90; 54%). In these age groups the prevalence of Fasciola was 28% and 39%, respectively, and for Paramphistomum the prevalence was 78% and 82%, respectively. Fifty percent of the calves younger than 3 months (n=68) were positive for Giardia, and none for Cryptosporidium. Most Giardia isolates were identified as the non-zoonotic G. duodenalis assemblage E on the beta-giardin gene. The blood samples were examined with commercially available Svanovir((R))Elisa's for the presence of Anaplasma marginale and Babesia bigemina specific antibodies, and a prevalence of 28% and 54% was found, respectively. In the milk samples Neospora caninum specific antibodies (Svanovir((R))Elisa) were detected in 30% of the lactating animals. The present study demonstrates that parasitic infections occur frequently in dairy cattle around Hanoi although animals are mainly kept confined, and indicates that further research on the economic impact of these infections is needed.