Assessment of anthelmintic efficacy against cattle gastrointestinal nematodes in western France and southern Italy.

Our objective was to measure the efficacy of ivermectin (IVM) and benzimidazoles (BZ, i.e. fenbendazole and albendazole) in 15 cattle farms in western France and southern Italy. A total of 11 groups were treated with IVM and 11 with BZ. Efficacy was assessed by calculating the percentage of faecal egg count reduction (%FECR) using the pre- and post-treatment arithmetic means. Anthelmintic resistance was considered to be present when the %FECR was <95% and the lower limit of the 95% confidence interval <90%. For IVM, the percentages of FECR ranged from 73% to 100%. Lack of efficacy to IVM was detected in two farms out of four in France, but was not detected in any of the seven farms in Italy. For BZ, the percentages of FECR ranged from 95% to 100%. No case of BZ resistance was detected in the five farms in France and the six farms in Italy.

A large outbreak of bovine botulism possibly linked to a massive contamination of grass silage by type D/C Clostridium botulinum spores on a farm with dairy and poultry operations.

Type D bovine botulism outbreaks associated with poultry litter are increasingly reported in European countries, but the circumstances of exposure to Clostridium botulinum toxins remain unclear. In spring 2015, a large type D/C bovine botulism outbreak affected a farm with dairy and poultry operations. Epidemiological and laboratory investigations strongly suggest that the outbreak was caused by feeding cattle with insufficiently acidified grass silage that was contaminated by type D/C C. botulinum spores. The source of the spores remains unclear, but could have been a stack of poultry litter stored in the grass silage pasture before harvesting. The presence of putrefied poultry carcasses mixed in with the litter is relatively unlikely considering the careful daily removal of poultry carcasses. These findings reinforce the importance of proper ensiling of feed materials and highlight the need for safe disposal of poultry litter, even in the case of good management of poultry deadstock, in order to prevent bovine botulism.

Molecular characterization of Cryptosporidium isolates from high-excreting young dairy calves in dairy cattle herds in Western France.

Ninety-two Cryptosporidium sp.-positive fecal samples of dairy diarrheic or non-diarrheic calves from 30 cattle herds in Normandy (France) were selected. Here, the aim was to investigate the species of Cryptosporidium excreted as well as the subtypes of Cryptosporidium parvum found in 7-17-day-old dairy calves. Excretion levels were comprised between 2 × 10(4) and 4 × 10(7) oocysts per gram of feces. Here, a nested 18S SSU rRNA PCR associated with sequencing was performed for identification of Cryptosporidium species and revealed the presence of C. parvum in most cases (80/82), except for two animals which were infected with Cryptosporidium bovis. Then, C. parvum samples were submitted to gp60 PCR. For 39 samples from 24 different herds, a multilocus analysis based on four mini-microsatellites loci (MM19, MM5, MSF, and MS9-Mallon) were conducted. These results were combined with sequence analysis of the gp60 to obtain multilocus types (MLTs). Here, C. parvum gp60 genotyping identified three subtypes in the IIa zoonotic allele family: IIaA15G2R1 (88%), IIaA16G3R1 (10%), and IIaA19G2R1 (2%), and we identified 12 MLTs. The MS9-Mallon locus was reported as the most polymorphic (five alleles). The most common MLT was MLT 1 with 15 samples in 10 farms: (MS9-M: 298, MSF: 165, MM5: 264, MM19: 462, and gp60 subtype: IIaA15G2R1). When comparing diarrheic and non-diarrheic fecal samples, no difference was seen for distribution of Cryptosporidium species, C. parvum gp60 subtypes, and MLTs. Here, in a range of oocyst excretion of 10(4)-10(7) opg, both in diarrheic and non-diarrheic calves, infection was mainly due to C. parvum and to the zoonotic subtype: IIaA15G2R1.

Fasciolosis in the Mediterranean island of Corsica (France): Insights from epidemiological and malacological investigations.

Fasciolosis is a re-emergent parasitic disease of worldwide significance with a major global impact on livestock health and production. In the French Mediterranean island of Corsica, fasciolosis has been recognized for a long time but little is known about its dynamic as the main investigations are outdated. Three compartments - definitive domestic hosts, intermediate hosts and environment - involved in fasciolosis transmission were studied by applying an integrative and extensive approach: (1) farm and abattoir surveys, (2) snail sampling, identification and infection prospection, and (3) snail habitat analysis; and (4) a questionnaire-based survey to inquire about husbandry practices and environmental risks. Our results indicate a significant circulation of the liver flukes in Corsican livestock, with 90% (252/279) of the sampled farms testing positive for anti-F. hepatica antibodies. At the abattoir, 46% (67/149) of cattle were positive for F. hepatica antibodies and eggs were present in the bile of 19% (26/139) bovines. In addition, high prevalence of Dicrocoelium dendriticum (69%) was observed in slaughtered cattle. Malacological surveys registered the occurrence of several lymnaeid species in a variety of habitats throughout the island. In particular, we report for the first time the presence of the invasive lymnaeid snail Pseudosuccinea columella in Corsica, a potential intermediate host for F. hepatica. We also found that the presence of Galba truncatula and, to a lesser extent, that of Peregriana peregra, is associated with altitude. Fasciola hepatica DNA was detected in the latter species occurring at two different sites. Finally, a questionnaire-based study revealed risky management practices among Corsican farmers, low perception of transmission and a suboptimal use of flukicide treatments as main control strategy. Our results show that animal fasciolosis in Corsica is characterised by a significant circulation and a favourable epidemiological scenario for transmission to occur.

Administration of the nematophagous fungus Duddingtonia flagrans to goats: an evaluation of the impact of this fungus on the degradation of faeces and on free-living soil nematodes.

The environmental impact of Duddingtonia flagrans, a potential biological control agent for nematode parasites, was tested in a 2-year-plot study using goat faeces. The trial assessed the impact of fungal presence on the disintegration of faeces and on non-target, free-living soil nematode populations. Three groups of goats experimentally infected by Trichostrongylus colubriformis received three different doses of D. flagrans chlamydospores (0 chlamydospores/kg body weight (BW), 0.5 × 10(6) chlamydospores/kg BW or 5 × 10(6) chlamydospores/kg BW). One hundred grams of faeces containing T. colubriformis eggs and D. flagrans chlamydospores at three different concentrations were deposited on pasture plots on four different occasions: May 2003, September 2003, June 2004 and September 2004. Faeces were weighed 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20 weeks after deposit and immediately afterwards replaced to their initial positions. In addition, soil samples were taken just below faecal deposits to evaluate the impact of fungal presence on non-target free-living nematodes. Results showed that there was no treatment effect on the pellet degradation rate. Analysis of soil nematode fauna failed to demonstrate any effect of the dose rate of 0.5 × 10(6) chlamydospores/kg BW, while a reduction of the number of free-living nematodes was seen for the maximal chlamydospore concentration at autumn sets.

Cryptosporidiosis in small ruminants.

Cryptosporidiosis is an infection caused by protozoan parasites belonging to the genus Cryptosporidium which is responsible for a potentially severe disease in new-born ruminants. This infection is highly prevalent in small ruminants throughout the world, especially in pre-weaned animals. The clinical expression is different between goat kids and lambs, the infection being generally more severe in the former. Molecular data demonstrate geographical variations in the species of Cryptosporidium infecting small ruminants. They also support the possibility of transmission of zoonotic species from these hosts to humans. Studies are still needed on molecular epidemiology, especially in goats, and on ways to control infection.

Gastrointestinal nematode and lungworm infections in organic dairy calves reared with nurse cows during their first grazing season in western France.

The rearing system of dairy calves with nurse cows has been developing since 2010 in organic farms in western France. This system allows cow-calf contact until a weaning age close to the natural weaning for cattle and is characterized by an early turnout for calves at around one month of age with their nurse cows and a first grazing season with mixed grazing of calves and adults at a ratio of 2-4 calves per nurse cow. The objectives of this study were to assess the gastrointestinal (GIN) and lungworm infections in such reared calves and their variability during the first grazing season. Faecal egg count (FEC), pepsinogen (PEP) concentration and Ostertagia ELISA optical density ratio (ODR) were determined in calves (n = 497) at housing in 33 groups from 24 farms in 2018, and in calves (n = 405) and nurse cows (n = 199) throughout the 2019 grazing season in 41 groups from 20 farms. For lungworm infection, information was obtained during 2019 through the recording of coughing episodes along the grazing season and the Dictyocaulus ELISA ODR determination at housing both in calves and nurses. Results indicated that the level of GIN infection was overall low for calves during the first grazing season with PEP and Ostertagia ODR group-average values ranging from 0.97 to 1.6 U Tyr and 0.23 to 0.71 ODR respectively. No anthelmintic treatment being given in any group of calves. Ostertagia ODR values increased with the duration of the grazing season (>240 d) and with the ratio calves/nurse (>2). GIN parameters for nurses remained fairly stable during the grazing season with mean FEC, PEP and Ostertagia ODR group-average values of 13 epg, 2.28 U Tyr and 0.81 ODR, respectively. Antibodies against lungworms were detected in 3-62 % of calves depending on the duration of grazing, but only 6% of calves showed a coughing episode. The dilution effect due to the mixed grazing of resistant (nurse cows) and susceptible (calves) animals associated with predominant milk diet of calves during the first months of grazing in combination with protective grazing management allow calves to be turned out at an early age without using anthelmintic treatments. Further studies are needed to assess the GIN infection dynamics during the second grazing season in weaned heifers.

A qualitative risk assessment methodology for scientific expert panels.

Risk assessment can be either quantitative, i.e. providing a numeric estimate of the probability of risk and the magnitude of the consequences, or qualitative, using a descriptive approach. The French Agency for Food, Environmental and Occupational Health and Safety (ANSES), formerly the French Food Safety Agency (AFSSA), bases its assessments on the opinions of scientific panels, such as the ANSES Animal Health Scientific Panel (AH-SP). Owing to the lack of relevant data and the very short period of time usually allowed to assess animal health risks on particular topics, this panel has been using a qualitative risk method for evaluating animal health risks or crises for the past few years. Some experts have drawn attention to the limitations of this method, such as the need to extend the range of adjectives used for the lower probabilities and to develop a way to assess consequences. The aim of this paper is to describe the improved method now established by the AH-SP, taking into account the limitations of the first version. The authors describe a new set of levels for probabilities, as well as the items considered when addressing either animal or human health consequences.

Rearing system with nurse cows and risk factors for Cryptosporidium infection in organic dairy calves.

Rearing dairy calves with nurse cows has been increasingly adopted by French farmers especially in organic farming and is characterized by a fostering of two to four calves during the first month of life by an unmilked lactating cow. This type of rearing remains poorly documented regarding its impact on calf health, such as cryptosporidiosis. The objectives of our study were to describe practices related to rearing dairy calves with nurse cows and to evaluate the prevalence, intensity and risk factors for Cryptosporidium infection in calf neonates. Between January and September 2019, the rearing practices of calves were described in 20 organic French farms and faeces were sampled once from 611 animals aged between 5 and 21 days. Cryptosporidium oocyst shedding was identified by modified Ziehl-Neelsen technique and scored semi-quantitatively (score 0-4). The risk of excretion (score 0 versus 1-4) was analysed using multivariate logistic regression models. This cow-calf rearing system usually consisted of a first phase with the dam, followed by an optional phase of artificial milk feeding (calves being fed with whole milk of the farm) and a final phase of fostering by a nurse cow. Each nurse was suckled from one to five calves of close age with a fostering age of 8 days on average. The oocyst shedding prevalence was 40.2 % and similar to classically reared calves, but the intensity of shedding and the prevalence of diarrhoea appeared to be lower. The identified six risk factors for oocyst shedding were: born in the last two thirds of the birth order, born between January and July versus August and September, calf with its dam in the barn versus on pasture, having an artificial milk feeding phase versus being with the dam only, and contact between peer calves and notably the presence of an oocyst excretory calf fostered by the same nurse. These results emphasize the role of the environment for the direct and indirect contamination, particularly that related to the accumulation of oocysts from previous or peer calves facilitating the faecal-oral route of transmission. This highlights the crucial role of the premises used intensively during the winter and spring months with higher densities of calves in the barn compared to outdoor situations promoted by this rearing.

Caprine nodular thelitis due to Mycobacterium uberis: A series of 26 cases in 11 dairy goat farms in Western France.

Bovine Nodular Thelitis (BNT) is a granulomatous dermatitis of teat skin associated with acid-fast bacilli. A similar condition has been recorded in a dairy goat flock in France recently. The causative agent was shown to be related to the leprosy-causing bacilli Mycobacterium leprae and M. lepromatosis, then sequenced and named M. uberis. Following the initial report in goats, the aim of this study was to investigate new cases of Caprine Nodular Thelitis (CNT) in the same area to confirm the presence of M. uberis by molecular techniques and to get a better description of the clinical signs and of the affected flocks. Twenty-six animals (25 females and 1 male) from 11 flocks were included in the study. Lesions were located on the udder/teat skin (24/25), on the body skin (6/25) or on the scrotum skin (1/1). Udder skin lesions were circular, nodular and/or ulcerate covered with a crust and associated with supramammary lymph node enlargement. Body skin lesions were located at different parts of the body, showed large necrotizing ulcers with undetermined edges and were associated with regional lymph node enlargement. Histopathological results indicated granulomatous dermatitis and lymphadenitis of varying intensity with no acid-fast bacilli seen after Fite-Faraco staining. M. uberis DNA was amplified from 26 samples out of 47 (udder: 11/22; lymph node: 11/20; body: 4/5). The female goats were mostly older than 4 year of age and originated from breeding units characterized by large flock size and high proportion of goat in continuous lactation.

Initial assessment of the economic burden of major parasitic helminth infections to the ruminant livestock industry in Europe.

We report a European wide assessment of the economic burden of gastrointestinal nematodes, Fasciola hepatica (common liver fluke) and Dictyocaulus viviparus (bovine lungworm) infections to the ruminant livestock industry. The economic impact of these parasitic helminth infections was estimated by a deterministic spreadsheet model as a function of the proportion of the ruminant population exposed to grazing, the infection frequency and intensity, the effect of the infection on animal productivity and mortality and anthelmintic treatment costs. In addition, we estimated the costs of anthelmintic resistant nematode infections and collected information on public research budgets addressing helminth infections in ruminant livestock. The epidemiologic and economic input data were collected from international databases and via expert opinion of the Working Group members of the European Co-operation in Science and Technology (COST) action COMbatting Anthelmintic Resistance in ruminants (COMBAR). In order to reflect the effects of uncertainty in the input data, low and high cost estimates were obtained by varying uncertain input data arbitrarily in both directions by 20 %. The combined annual cost [low estimate-high estimate] of the three helminth infections in 18 participating countries was estimated at € 1.8 billion [€ 1.0-2.7 billion]. Eighty-one percent of this cost was due to lost production and 19 % was attributed to treatment costs. The cost of gastrointestinal nematode infections with resistance against macrocyclic lactones was estimated to be € 38 million [€ 11-87 million] annually. The annual estimated costs of helminth infections per sector were € 941 million [€ 488 - 1442 million] in dairy cattle, € 423 million [€ 205-663 million] in beef cattle, € 151million [€ 90-213 million] in dairy sheep, € 206 million [€ 132-248 million] in meat sheep and € 86 million [€ 67-107 million] in dairy goats. Important data gaps were present in all phases of the calculations which lead to large uncertainties around the estimates. Accessibility of more granular animal population datasets at EU level, deeper knowledge of the effects of infection on production, levels of infection and livestock grazing exposure across Europe would make the largest contribution to improved burden assessments. The known current public investment in research on helminth control was 0.15 % of the estimated annual costs for the considered parasitic diseases. Our data suggest that the costs of enzootic helminth infections which usually occur at high prevalence annually in ruminants, are similar or higher than reported costs of epizootic diseases. Our data can support decision making in research and policy to mitigate the negative impacts of helminth infections and anthelmintic resistance in Europe, and provide a baseline against which to measure future changes.

Efficacy of oxyclozanide against the rumen fluke Calicophoron daubneyi in experimentally infected goats.

Paramphistome infections are very common in ruminants and may induce clinical signs, but little is known about effective treatments. In this study, the efficacy of oxyclozanide against Calicophoron daubneyi was studied in goats and its activity tested against immature stages (10 days post-infection) at a dose of 22.5mg/kg bodyweight (BW) and against adult stages using two doses (15 and 22.5mg/kg BW). There was a reduction (82%) in the number of immature worms (compared to controls) but the result was not statistically significant. When tested against adult stages, however, oxyclozanide reduced the worm burdens by 95.6% and 95.9% at doses of 15 and 22.5mg/kg BW, respectively, with no significant difference between the two doses. The experiment demonstrated that oxyclozanide is highly effective in reducing the number of adult paramphistomes in goats.

Mid-season targeted selective anthelmintic treatment based on flexible weight gain threshold for nematode infection control in dairy calves.

The suitability of a single mid-season targeted selective treatment (TST) for gastrointestinal nematodes control, based on flexible average daily weight gain (ADWG) thresholds, was investigated in 23 groups of first grazing season calves. In each group, animals were weighed three times: before turnout, at mid-season and at housing. Just after the first weighing, each group was divided in two homogenous sub-groups in terms of age, breed and weight, and randomly allocated to one of two sub-groups intented for two different mid-season anthelmintic treatment strategies: (1) a treatment of all calves composing the sub-group (whole-group treatment (WT)) or (2) a targeted selective weight gain-based treatment (TST) of the animals showing an individual pre-treatment ADWG inferior to the mean pre-treatment ADWG of the corresponding WT sub-group. Anthelmintic treatment (levamisole 7.5 mg/kg BW) was performed 3 to 4 months after turnout. At housing, two parasitological parameters (the anti-Ostertagia ostertagi antibody level-Ostertagia optical density ratio (ODR) and the pepsinogen level) and a clinical parameter (the breech soiling score) were assessed at individual level in each group. Then, the high exposed groups to gastrointestinal nematode (GIN) were defined as groups for which untreated animals exhibited a mean Ostertagia ODR ⩾0.7 and among these groups, the ones characterized by high abomasal damage due to Ostertagia for which untreated animals exhibited a mean pepsinogen level ⩾2.5 U Tyr were also identified. Among TST sub-groups, the treatment ADWG thresholds varied from 338 to 941 g/day and the percentage of treated animals from 28% to 75%. Pre- and post-treatment ADWG as well as parasitological and clinical parameters measured at housing were similar between TST and WT sub-groups including the 17 high exposed groups to GIN. Within these 17 groups, the treatment allowed to significantly improve post-treatment ADWG compared with untreated animals. In the six high exposed groups showing mean pepsinogen level ⩾2.5 U Tyr, the average effect of treatment on post-treatment ADWG was the highest and estimated up to 14 kg after a grazing duration of 4 months. In contrast, in six other groups showing mean Ostertagia ODR<0.7 in untreated animals, no effect of treatment was seen suggesting an absence of production losses related to a low level of GIN infection. This study highlighted the suitability of a convenient mid-season TST strategy for first grazing season calves, based on the use of flexible thresholds of ADWG, allowing similar growth compared with a whole-group treatment while keeping a GIN population in refugia.

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.

Efficiency of feeding Duddingtonia flagrans chlamydospores to control nematode parasites of first-season grazing goats in France.

A field trial, conducted over two consecutive years, was aimed at assessing the efficacy of the administration of spores of the nematophagous fungus Duddingtonia flagrans to young goats for the control of nematode parasite infections on a French commercial dairy goat flock. For both years, the first-year grazing kids were divided into two similarly managed groups (fungus and control groups): in 2003 a daily dose rate of 5 x 10(5) spores/kg body weight was given to the fungus-group animals, while in 2004 a daily dose rate of 10(6) spores/kg body weight was used; the other half of the kids, acting as control, did not receive the spores. Parameters measured every 3 weeks included nematode egg excretion, larval development in faecal cultures and pasture larval counts. Additionally, at the beginning, the middle and the end of each grazing season, the goats were weighed and blood samples for pepsinogen determination were collected. In 2003, similar results were recorded for all the measured parameters in the control and fungus groups. In contrast, in 2004, the kids receiving the spores showed lower faecal egg counts and pepsinogen levels at the end of the season and higher growth rate compared to kids of the control group.

Pharmacokinetics and anthelmintic efficacy of injectable eprinomectin in goats.

The objective of this study was to assess the pharmacokinetics and the anthelmintic efficacy of eprinomectin (Eprecis® 20mg/mL) following subcutaneous administration to goats. Forty non-lactating female Alpine goats aged between one and three years and weighing between 32.7 and 59.5kg, were randomly allocated to one of the following groups (8 animals per group): two groups were not infected and were treated at a dose of either 0.2 or 0.4mg/kg BW of eprinomectin, two groups were experimentally infected with nematodes and treated at 0.2 or 0.4mg/kg BW of eprinomectin similarly and one group was infected and left untreated (control). Infection consisted in a single and simultaneous administration of 5000 Haemonchus contortus and 12,500 Trichostrongylus colubriformis infective larvae. No local or general adverse reaction was visually observed whatever the dose rate. The maximal plasma concentrations (Cmax) were 20.68±12.85 vs 39.79±17.25µg/L and the plasma bioavailabilities (AUC) 83.45±34.75 vs 169.37±43.44µg*d/L for 0.2 vs 0.4mg/kg respectively. The efficacy against H. contortus and T. colubriformis was 97.8 and 98.7% at 0.2mg/kg and 98.4% and >99.9% at 0.4mg/kg respectively. The differences in worm burdens between the two dose rates were only significant for T. colubriformis. These results indicate that injectable eprinomectin is a potent anthelmintic against the two major gastrointestinal nematodes in goats. Additional information is needed regarding pharmacokinetics in lactating goats and milk residues.

Comparative efficacy of the nematode-trapping fungus Duddingtonia flagrans against Haemonchus contortus, Teladorsagia circumcincta and Trichostrongylus colubriformis in goat faeces: influence of the duration and of the temperature of coproculture.

Amongst the alternative strategies to the use of anthelmintics, the administration of Duddingtonia flagrans spores has already proved its efficacy in reducing the number of developing larvae of several nematode species in goat faeces. In this trial, the efficacy of this fungus against the larvae of the three major nematode species of goats was compared in various conditions of coproculture. Twelve strongyle free goats were experimentally infected with either Trichostrongylus colubriformis, Teladorsagia circumcincta or Haemonchus contortus larvae. Half of the animals received an oral dose of 5x10(5) Duddingtonia chlamydospores/kg BW daily for 27 days, whereas the remaining was kept as control goats. From the 7th day of administration onwards, individual coproscopical examinations as well as coprocultures, which were incubated 4, 7, 10 or 14 days at 21 or 28 degrees C, were performed. The reduction in developing larvae due to the activity of Duddingtonia ranged from 62.8 to 99.5% compared to control. The trapping efficacy depended on temperature (better activity of the fungus at 21 than at 28 degrees C) and on duration (larval reductions lower after 4 days than after 7, 10 and 14 days of coproculture). Teladorsagia larvae were the least trapped, and Haemonchus larvae were the most trapped.