Evaluation of the flukicide treatment policy for dairy cattle in Galicia (NW Spain).

Fasciola hepatica infection is an important cause of lost productivity in livestock worldwide. Effective control of fasciolosis is difficult, especially in milking cows, which can only be treated during dry periods, a control strategy that has not been yet evaluated. In this cross-sectional study, we investigated the effect of the type of flukicide treatment on the prevalence and intensity of infection in dairy cattle from Galicia, an area where fasciolosis is endemic and which is also the main milk-producing region in Spain. Faecal samples were taken from 5188 dairy cows on 275 randomly selected farms for measurement of the concentration of F. hepatica coproantigens by a monoclonal antibody based immunoassay (MM3-COPRO ELISA). On the same day as the sampling, each farm owner/manager was questioned about the types of treatment used on the farm. Three groups of farms were considered according to the fasciolicide treatment: (A) flukicides were not used, (B) an anthelmintic effective against mature stages of flukes was used (albendazole or netobimin) and (C) a fasciolicide effective against immature and mature stages was used (triclabendazole: TCBZ). Results indicated that 16.0% (832/5188) cows from 61.1% (168/275) herds were infected by F. hepatica. The mean coproantigen concentration in infected herds was 13.0ng/ml (range 0.9-112.6ng/ml). The highest individual concentration recorded was 496.6ng/ml. Herd and within-herd prevalences of F. hepatica were similar in all three groups, but surprisingly, individual prevalence and antigen concentration were higher in Group C (p<0.05). The percentage of farms with within-herd prevalences >25% was very high in all three groups, and no significant differences were observed. In contrast, the percentage of herds with mean antigen concentrations >20ng/ml was significantly lower (p<0.05) in Groups A and B (14.4% and 14.9%, respectively) than in Group C (50.0%). The proportion of herds that exceeded both limits (25% for prevalence and/or 20ng/ml for coproantigen concentration) was also significantly higher (p<0.05) in Group C than in untreated animals (Group A). The survey showed that most dairy farmers are unaware of the existence of F. hepatica infection on their farms, and treatments, when given, are administered without prior diagnosis. Treatment with TCBZ administered only at drying off did not show advantages over other measures including no treatment, or treatment with other benzimidazoles. Consequently, TCBZ should only be used to treat individual animals after correct diagnosis of the infection, and correct management measures taken to control re-infection.

Mezo M ，González-Warleta M ，Castro-Hermida JA ，Ubeira FM ... -  《VETERINARY PARASITOLOGY》

High prevalence of fasciolosis and evaluation of drug efficacy against Fasciola hepatica in dairy cattle in the Maffra and Bairnsdale districts of Gippsland, Victoria, Australia.

Liver fluke (Fasciola hepatica) is a common parasite amongst grazing livestock in the south-eastern region of Australia and is responsible for significant production losses in the beef and dairy industries. Gippsland in Victoria is a major region for dairy production but no fluke prevalence data in livestock has been obtained in this region since the late 1970s prior to the introduction of Triclabendazole (TCBZ). TCBZ resistance is also now widespread in cattle in south east Australia. In this study, we evaluated the prevalence and intensity of liver fluke infections in dairy cattle in Gippsland and assessed the efficacy of TCBZ and other drenches against F. hepatica on one farm. We obtained 30 individual faecal samples from each of 15 different farms and, using the liver fluke coproantigen ELISA, tested bulk faecal samples pooled from each farm. Any farm that returned a positive bulk sample had all of the samples tested individually to assess the intra-herd prevalence. One farm in the Maffra district also had a coproantigen reduction test and faecal egg count reduction test to assess the efficacy of TCBZ, Clorsulon (CLOR) and Oxyclozanide (OXY). The coproantigen ELISA proved to be a highly sensitive test for liver fluke with a high correlation (R(2)=0.8849) observed between ELISA data from bulk samples and individual samples, suggesting that future larger scale screening on farms for fasciolosis could use the bulk analysis technique. The ELISA data revealed that animals on six of the 15 farms were infected with F. hepatica and the herd prevalence of the infected herds ranged from 47 to 100% (mean 81%) which exceeds the prevalence value for production losses of 25%. The intensity of fluke infection in cattle varied considerably both within and between herds with a proportion of animals exhibiting a positive control value in the coproantigen ELISA of 50-88%. We also confirmed that TCBZ resistance was present on one farm but that CLOR or OXY can be used to remove the adult stage of the TCBZ-resistant parasites. We conclude that fasciolosis is a significant disease and a likely cause of production losses in dairy cattle in the irrigation zones of Gippsland and that TCBZ resistance is a serious threat to fluke control. We suggest that more work needs to be performed in Gippsland to further define the extent of fasciolosis and drug resistance and to ensure that effective chemical and non-chemical methods of fluke control are incorporated on farms in order to improve animal welfare and reduce financial impacts on producers.

Elliott TP ，Kelley JM ，Rawlin G ，Spithill TW ... -  《-》

First report of closantel treatment failure against Fasciola hepatica in cattle.

Control of Fasciola hepatica infection in livestock is based on annual treatment using flukicides such as triclabendazole, albendazole and closantel. However, triclabendazole resistant F. hepatica populations are emerging worldwide and resistance is emerging to albendazole, whereas it has until now never been described for closantel. In Sweden, a topical formulation containing a combination of closantel and ivermectin (Closamectin Pour On) has been registered for use in cattle only since 2011. This study evaluated the efficacy of closantel against F. hepatica in naturally infected beef cattle using both coproantigen and faecal egg count reduction tests. Faecal egg counts (FEC) and coproantigen ELISA examinations were conducted in February 2014 in three beef cattle herds (A, B, C) in south-western Sweden. On each farm, 10 F. hepatica coproantigen-positive and F. hepatica egg-positive animals were allocated after 12-16 weeks of housing into groups and treated topically with a minimum of 20 mg closantel per kg body weight. Faecal samples were collected from selected animals on 0, 7 and 21 day post-treatment (PT). Based on FEC, closantel efficacy 21 days PT was 72% (95% CI: 65-77%) and 97% (95% CI: 95-98%) on farms A and B, respectively. No FEC reduction at all was observed on farm C. In total, 4, 1 and 6 animals remained coproantigen-positive at 21 days PT on farms A, B and C, respectively. Closantel treatment failure was confirmed on two of the farms. As the animals were housed 12-16 weeks before treatment and thereafter during the entire study, failure due to the presence of juvenile flukes was excluded. Although the cause of closantel failure currently remains unclear, development of resistance or/and absorption failure of topical administration should be considered. To our knowledge, this is the first report of closantel treatment failure against F. hepatica in cattle.

Novobilský A ，Höglund J 《International Journal for Parasitology-Drugs and Drug Resistance》

The effects of farm management practices on liver fluke prevalence and the current internal parasite control measures employed on Irish dairy farms.

Fasciolosis caused by Fasciola hepatica is responsible for major production losses in cattle farms. The objectives of this study were to assess the effect of farm management practices on liver fluke prevalence on Irish dairy farms and to document the current control measures against parasitic diseases. In total, 369 dairy farms throughout Ireland were sampled from October to December 2013, each providing a single bulk tank milk (BTM) sample for liver fluke antibody-detection ELISA testing and completing a questionnaire on their farm management. The analysis of samples showed that cows on 78% (n=288) of dairy farms had been exposed to liver fluke. There was a difference (P<0.05) between farms where cows were positive or negative for liver fluke antibodies in (a) the total number of adult dairy cows in herds, (b) the number of adult dairy cows contributing to BTM samples, and (c) the size of the total area of grassland, with positive farms having larger numbers in each case. There was no difference (P>0.05) between positive and negative farms in (a) the grazing of dry cows together with replacement cows, (b) whether or not grazed grassland was mowed for conservation, (c) the type of drinking water provision system, (d) spreading of cattle manure on grassland or (e) for grazing season length (GSL; mean=262.5 days). Also, there were differences (P<0.001) between drainage statuses for GSL with farms on good drainage having longer GSL than moderately drained farms. The GSL for dairy cows on farms with good drainage was 11 days longer than for those with moderate drainage (P<0.001). The percentage of farmers that used an active ingredient during the non-lactating period against liver fluke, gastrointestinal nematodes, lungworm, and rumen fluke was 96%, 85%, 77% and 90%, respectively. Albendazole was the most frequently used active ingredient for treatment against gastrointestinal nematodes (57%), liver fluke (40%) and lungworm (47%), respectively. There was a difference (P<0.05) in the use of triclabendazole and albendazole between positive and negative farms, with triclabendazole use being more common in positive farms. This study highlighted differences in dairy management practices between Irish farms with dairy herds exposed or not exposed to liver fluke and stressed the need of fine-scale mapping of the disease patterns even at farm level to increase the accuracy of risk models. Also, comprehensive advice and professional support services to farmers on appropriate farm management practices are very important for an effective anthelmintic control strategy.

Selemetas N ，Phelan P ，O'Kiely P ，de Waal T ... -  《-》

An ultrasensitive capture ELISA for detection of Fasciola hepatica coproantigens in sheep and cattle using a new monoclonal antibody (MM3).

A capture enzyme-linked immunosorbent assay (ELISA) using a new monoclonal antibody (mAb MM3) is reported for the detection of Fasciola hepatica excretory-secretory antigens (ESAs) in feces of infected hosts. The mAb MM3 was produced by immunization of mice with a 7- to 40-kDa purified and O-deglycosylated fraction of F. hepatica ESAs, which has previously been shown to be specific for the parasite. The specificity and sensitivity of the MM3 capture ELISA were assessed using feces from sheep and cattle. Sheep feces were obtained from a fluke-free herd (with most animals harboring other nematodes and cestodes), from lambs experimentally infected with 5-40 F. hepatica metacercariae and in some cases treated with triclabendazole at 14 wk postinfection (PI), and from uninfected control lambs. Cattle feces were collected at the slaughterhouse from adult cows naturally infected with known numbers of flukes (from 1 to 154) or free of F. hepatica infection (though in most cases harboring other helminths). The MM3 capture ELISA assay had detection limits of 0.3 (sheep) and 0.6 (cattle) ng of F. hepatica ESA per milliliter of fecal supernatant. The assay detected 100% of sheep with 1 fluke, 100% of cattle with 2 flukes, and 2 of 7 cattle with 1 fluke. The false-negative animals (5/7) were probably not detected because the F. hepatica individuals in these animals were immature (5-11 mm in length). As expected, coproantigen concentration correlated positively (r = 0.889; P < 0.001) with parasite burden and negatively (r = 0.712; P < 0.01) with the time after infection at which coproantigen was first detected. Nevertheless, even in animals with low fluke burdens (1-36 parasites), the first detection of F. hepatica-specific coproantigens by the MM3 capture ELISA preceded the first detection in egg count by 1-5 wk. In all sheep that were experimentally infected and then untreated, coproantigen remained detectable until at least 18 wk PI, whereas in sheep that were experimentally infected and then flukicide treated, coproantigen became undetectable from 1 to 3 wk after treatment. None of the fecal samples from sheep or cattle negative for fascioliasis but naturally infected with other parasites including Dicroelium dendriticum showed reactivity in the MM3 capture ELISA. These results indicate that this assay is a reliable and ultrasensitive method for detecting subnanogram amounts of F. hepatica antigens in feces from sheep and cattle, facilitating early diagnosis.

Mezo M ，González-Warleta M ，Carro C ，Ubeira FM ... -  《JOURNAL OF PARASITOLOGY》