Gastrointestinal nematodes and Fasciola hepatica in Norwegian cattle herds: a questionnaire to investigate farmers' perceptions and control strategies.

BACKGROUND: Pasture management influences the prevalence and impact of the pasture parasites (PP) in cattle herds, which cause production-limiting disease worldwide. Evaluating farmer management strategies is vital when considering sustainable PP control practices. The aim of this questionnaire-based study was to describe the pasture management and control strategies regarding PP in Norwegian beef cattle (BC) and dairy cattle (DC) production systems with a focus on gastrointestinal nematodes (GIN) and Fasciola hepatica. RESULTS: A total of 745 responses from BC (return rate 20.5%) and 1347 responses from DC farmers (30.7%) were included. The mean total pasture time for DC was 4.2 months for first-season grazers and 4.3 months for second-season grazers and cows, while the corresponding finding in BC was 5.4 months. Home pasture was used for most of the pasture period, particularly for first-season grazer dairy heifers (81%), which were also commonly grazed on the same pasture every year (79%). For most farmers it was necessary for grazing areas to be used for cattle for more than one season (77% of BC farmers and 89% of DC farmers). However, changing the pasture during the season was common in both DC (67%) and BC (70%) herds. The majority of DC farmers (60%) stated that they did not consider that they had a problem with PP. Of the remaining 40%, few respondents could specify whether their herds had a problem due to infection by GIN (11%) or liver flukes (12%). Treatment for GIN was performed by 52% of DC and 34% of BC farmers. Diagnostic faecal samples were collected upon suspicion of parasitic disease by 5% of DC and 16% of BC farmers. Veterinarians were stated as a central source of information about parasite management and treatment. CONCLUSIONS: Potential risks for exposure to PP were identified, such as use of the same pasture every year for first-season grazers and frequent use of home pasture. The perception of problems related to PP appeared low. Regular anthelmintic treatment without concurrent use of diagnostic faecal samples seems to be common practice.

Use of Oxidative Stress Responses to Determine the Efficacy of Inactivation Treatments on Cryptosporidium Oocysts.

Cryptosporidium oocysts are known for being very robust, and their prolonged survival in the environment has resulted in outbreaks of cryptosporidiosis associated with the consumption of contaminated water or food. Although inactivation methods used for drinking water treatment, such as UV irradiation, can inactivate Cryptosporidium oocysts, they are not necessarily suitable for use with other environmental matrices, such as food. In order to identify alternative ways to inactivate Cryptosporidium oocysts, improved methods for viability assessment are needed. Here we describe a proof of concept for a novel approach for determining how effective inactivation treatments are at killing pathogens, such as the parasite Cryptosporidium. RNA sequencing was used to identify potential up-regulated target genes induced by oxidative stress, and a reverse transcription quantitative PCR (RT-qPCR) protocol was developed to assess their up-regulation following exposure to different induction treatments. Accordingly, RT-qPCR protocols targeting thioredoxin and Cryptosporidium oocyst wall protein 7 (COWP7) genes were evaluated on mixtures of viable and inactivated oocysts, and on oocysts subjected to various potential inactivation treatments such as freezing and chlorination. The results from the present proof-of-concept experiments indicate that this could be a useful tool in efforts towards assessing potential technologies for inactivating Cryptosporidium in different environmental matrices. Furthermore, this approach could also be used for similar investigations with other pathogens.

Occurrence of faecal endoparasites in reindeer (Rangifer tarandus) in two grazing areas in northern Norway.

BACKGROUND: Semi-domesticated reindeer represent an important livestock industry and livelihood for a proportion of the human population in northern Fennoscandia. Reindeer husbandry is considered an extensive animal husbandry, where the animals are kept mostly on natural pastures, although sometimes kept in fenced areas for shorter periods. These reindeer may harbour a variety of parasites that may affect animal health and production. The relatively limited close contact between herds and owners gives limited opportunities for diagnosis and treatment of diseases in general. Furthermore, the effects of subclinical parasitism in livestock are commonly expressed as a reduction in productivity rather than clinical disease and mortality. Thus, specific knowledge of endoparasites and parasitic infections in these herds is scarce. This study investigated the occurrence of various endoparasites in reindeer by analysis of a total of 114 faecal samples from winter-slaughtered reindeer from two different grazing areas in Troms and Finnmark, Norway. RESULTS: Using a McMaster method, a Baermann technique, and a direct immunofluorescent antibody test, the following parasites were identified in the faecal samples with the occurrence data given as percentages: Strongylid eggs (62%), Nematodirinae spp. eggs (24%), Capillaria sp. eggs (42%) and Moniezia spp. eggs (17%), Dictyocaulus spp. larvae (14%) protostrongylid larvae (40%), Eimera spp. oocysts (23%), and Giardia duodenalis cysts (5%). Cryptosporidium oocysts were not detected. Parasite eggs, oocysts, and cysts were quantified. Molecular analysis revealed G. duodenalis sub-assemblage AI, a potentially zoonotic genotype not previously reported in reindeer from this region. Morphological analyses of Eimeria oocysts identified two species, Eimeria mayeri and Eimeria rangiferis, and molecular analyses of the cytochrome C oxidase I (coi) gene and 18 s rRNA (18SSU) gene of Eimeria confirmed the presence of Eimeria species that are specific to reindeer. CONCLUSIONS: A high prevalence, but low burden, of endoparasites was detected in samples from these semi-domesticated reindeer. The samples were collected during winter, when adult gastrointestinal parasites usually produce only low numbers of transmission stages. Therefore, together with the low number of samples, detailed and definitive conclusions regarding parasite status of semi-domesticated reindeer are avoided. Nevertheless, these data provide a snapshot overview of parasite occurrence in a semi-domesticated animal group vulnerable to the various environmental changes to which they are exposed.

Pre- and Post-Race Intestinal Microbiota in Long-Distance Sled Dogs and Associations with Performance.

Although our understanding of the role of the gut microbiota in different diseases is improving, our knowledge regarding how the gut microbiota affects functioning in healthy individuals is still limited. Here, we hypothesize that the gut microbiota could be associated with sled dog endurance-race performance. We investigated the gut microbiota in 166 fecal samples from 96 Alaskan Huskies, representing 16 teams participating in the 2016 Femund Race (400 km) in Norway, relating the microbiota composition to performance and metadata derived from questionnaires. For 16S rRNA gene sequencing-derived compositional data, we found a strong negative association between Enterobacteriaceae (dysbiosis-associated) and Clostridium hiranonis (normobiosis-associated). The teams with the best performances showed both the lowest levels of dysbiosis-associated bacteria prior to the race and the lowest change (decrease) in these bacteria after the race. Taken together, our results support the hypothesis that normobiosis-associated bacteria are involved in resilience mechanisms, potentially preventing growth of Enterobacteriaceae during the race.

A novel multiplex real-time PCR for the detection of Echinococcus multilocularis, Toxoplasma gondii, and Cyclospora cayetanensis on berries.

Foodborne parasites (FBP) are of major public health importance and warrant appropriate detection and control strategies. Most of the FBP considered for risk-ranking by a panel of experts are potentially transmitted via consumption of contaminated fresh produce, including berries. In this study we focused on the potential of three FBP, namely Echinococcus multilocularis, Toxoplamsa gondii, and Cyclospora cayetanensis, as contaminants of berries. Surveys to assess these parasites as contaminants of fresh produce in general, and berries in particular, are scanty or non-existent mainly due to the lack of optimized laboratory methods for detection. The aim of the present study was to develop and evaluate a novel multiplex qPCR for the simultaneous detection of E. multilocularis, T. gondii, and C. cayetanensis from berry fruits. The efficiency and linearity of each channel in the multiplex qPCR were within the acceptable limits for the range of concentrations tested. Furthermore, the method was shown to have good repeatability (standard deviation ≤0.2 Cq) and intermediate precision (pooled standard deviation of 0.3-0.6 Cq). The limit of detection was estimated to 10 oocysts for Toxoplasma and Cyclospora, and 5 eggs for Echinococcus per 30 g of raspberries or blueberries. In conclusion, evaluation of the present method showed that the newly developed multiplex qPCR is highly specific, precise, and robust method that has potential for application in food-testing laboratories.

Anoplocephala perfoliata Infection in Horses in Iceland: Investigation of Associations Between Intensity of Infection and Lesions.

In Iceland, there is at least 1 horse for every 5 human inhabitants, mostly kept on uncultivated rangelands. Although the Icelandic horse is considered robust compared with other breeds, it is nevertheless susceptible to disease. Few studies have investigated the prevalence of intestinal parasites in horses in Iceland. The purpose of this study was to determine the prevalence of the tapeworm, Anoplocephala perfoliata, in horses in Iceland and to explore associations between intensity of infection and the severity of macroscopic pathological lesions in this population. In addition, the relationship between infection and geographical location in Iceland, horse age, and gender were investigated. The utility of a modified McMaster flotation method for identifying infected horses from fecal samples was also studied. The study sample consisted of 104 horses (aged 1-30 yr) slaughtered in 3 abattoirs in North and South Iceland during June and July 2016. The prevalence of A. perfoliata in the horses was 64.4%, with between 1-131 worms found in infected horses. The pathological changes in the horses associated with infection varied from mild (hyperemia in small areas) to severe (large ulcers and necrotic lesions coated with fibrin), and intensity of infection correlated with the size and type of pathological lesions in the intestines. No statistically significant associations between infection with A. perfoliata and geographical location, horse age, or horse gender were detected. The McMaster egg counting method used here was of very low sensitivity (7.4%) and was concluded to be inadequate for detection of A. perfoliata infection in horses. The prevalence of A. perfoliata in horses in Iceland in this study was found to be relatively high compared with results from many studies performed elsewhere in Europe. This may reflect the lack of treatment of horses in Iceland for this parasite, the environmental suitability for the intermediate mite host, and grazing management practices in Iceland. The high prevalence and association with pathology suggest that diagnosis of infection using a sensitive method, along with appropriate treatment, should be considered for horses in Iceland.

Not just a walk in the park: Occurrence of intestinal parasites in dogs roaming recreational parks in Chandigarh, Northern India.

Chandigarh, a city in North-west India, has numerous parks and recreational areas where stray dogs roam freely and pet dogs are exercised. This allows for extensive human-dog interaction, which may pose a public health threat. The aim of this study was to determine the occurrence of gastrointestinal parasites of public health importance, and their seasonal variation, in canine faecal samples obtained from recreational parks in Chandigarh. A total of 212 samples were collected from 10 parks during the winter (January 2015; N = 107) and monsoon season (September 2015; N = 105), to assess the prevalence of intestinal zoonotic parasites and any seasonal variations. The samples were analysed for helminth eggs using McMaster counting chambers. Immunofluorescent antibody testing was used to analyse samples for Cryptosporidium oocysts and Giardia cysts. The Giardia-positive samples were genotyped by conventional multi-locus PCR to determine their assemblage and zoonotic potential. Among the 212 samples, strongyle-type eggs were found in 34 (16.0%), Toxocara spp. eggs were found in 6 (2.8%), taeniid eggs in 1 (0.5%), Cryptosporidium spp. oocysts in 4 (1.9%) and Giardia duodenalis cysts in 49 (23.1%). Trichuris eggs were not detected. The majority of the successfully amplified Giardia isolates belonged to canid-specific assemblages. The prevalence of Giardia cysts in faecal samples was significantly higher during winter than in the monsoon season, whereas helminth-egg prevalence unaffected by season. The prevalence of strongyle-type eggs and Giardia cysts in dog faeces was lower in more affluent areas of the city than those of less affluence. There was no significant difference in the intensity of infection between the seasons. The results indicate that faeces from dogs contaminating parks in Chandigarh do not usually contain parasite transmission stages that pose a significant risk to human health. However, the importance of minimising contamination of public parks with dog faeces is highlighted.

Lectin-magnetic separation (LMS) for isolation of Toxoplasma gondii oocysts from concentrated water samples prior to detection by microscopy or qPCR.

Although standard methods for analyzing water samples for the protozoan parasites Cryptosporidium spp. and Giardia duodenalis are available and widely used, equivalent methods for analyzing water samples for Toxoplasma gondii oocysts are lacking. This is partly due to the lack of a readily available, reliable immunomagnetic separation technique (IMS). Here we investigated the use of lectin-magnetic separation (LMS) for isolating T. gondii oocysts from water sample concentrates, with subsequent detection by microscopy or molecular methods. Four different types of magnetic beads coated with wheat germ agglutinin (WGA) were tested for capture of oocysts from clean or dirty water samples. Dynabeads (Myone T1 and M-280) consistently provided mean capture efficiencies from 1 ml clean water in excess of 97%. High recoveries were also found with Tamavidin beads (in excess of 90%) when LMS was used for capture from a small (1 ml) volume. Dissociation (required for detection by microscopy) using 0.1N hydrochloric acid (HCl), as standard in IMS, was not successful, but could be achieved using a combination of acidified pepsin (AP) and N-acetyl d-glucosamine. Although simple centrifugation was as effective as LMS when concentrating high numbers of oocysts from clean water, LMS provided superior results when oocysts numbers were low or the water sample was dirty. Application of LMS integrated with qPCR enabled detection of 10 oocysts per 10 ml dirty water sample concentrate. These findings indicate that LMS with WGA coupled to magnetic beads could be an efficient isolation step in the analysis of water sample concentrates for T. gondii oocysts, with detection either by microscopy or by qPCR.

Treatment of feline giardiasis during an outbreak of diarrhoea in a cattery: potential effects on faecal Escherichia coli resistance patterns.

OBJECTIVES: An outbreak of diarrhoea involving 16 cats at a cattery in Norway was investigated. Treatment and control of the outbreak were the primary objectives, but the effects of treatment on the antimicrobial resistance profiles of Escherichia coli isolated from faeces were also investigated. METHODS: Faecal samples were investigated for Giardia cysts by immunofluorescence microscopy, and multi-locus genotyping was performed to determine the Giardia genotype. Faecal E coli were assessed, before and after treatment for giardiasis, for antimicrobial resistance. RESULTS: The outbreak was probably caused by Giardia duodenalis, Assemblage F. Although infection was eliminated in most cats following treatment with fenbendazole, over 30% of the infected cats required a second treatment round (combined fenbendazole and metronidazole). Investigation of sensitivity to antibacterial drugs of E coli that had been isolated both prior to and following treatment demonstrated that fenbendazole treatment may select for resistant bacteria. CONCLUSIONS AND RELEVANCE: Controlling Giardia infections in dense cat populations can be challenging, and requires strict hygiene measures. In cases where fenbendazole alone does not result in treatment success, a combination treatment with fenbendazole and metronidazole may be effective. Although this study did not include untreated controls, we suggest that the potential for changes in gut microbiota and antimicrobial resistance development should be considered when choosing antiprotozoal drugs, particularly in cases of treatment failure and where repeat treatment is required.

Subclinical Giardia in dogs: a veterinary conundrum relevant to human infection.

Human infection with Giardia duodenalis tends to be associated with diarrheal disease requiring treatment - despite our awareness that often it is asymptomatic and sometimes, perhaps, even protective. We discuss here whether canine giardiasis can serve as a model to help to understand why Giardia is pathogenic. We discuss factors that should be considered when Giardia is identified in dogs, challenging the assumption that infection necessarily means disease that requires chemotherapeutic treatment. To make the best treatment decision for canine Giardia infection we need to think about zoonotic risks, transmission possibilities, and risk factors for disease development. In addition, in both humans and dogs, Giardia sometimes may be considered as a harmless passenger, or even as a beneficial friend.