• Comparison of four commercially available ELISA kits for diagnosis of Fasciola hepatica in Irish cattle

      Munita, Maria P; Rea, Rosemary; Martinez-Ibeas, Ana M; Byrne, Noel; Kennedy, Aideen; Sekiya, Mary; Mulcahy, Grace; Sayers, Riona; Department of Agriculture, Food and the Marine; Dairy Research Ireland; et al. (Biomed Central, 2019-11-21)
      Background Fasciola hepatica is a liver parasite of mammals and it results in poor welfare outcomes and economic losses in ruminants. While faecal egg count is the test most commonly used for diagnosis, it does not indicate presence of migrating immature stages. Serological techniques increase sensitivity at all stages of the liver fluke infection. The aim of this study was to compare four commercially available ELISA tests for the diagnosis of F. hepatica. For this purpose, we tested three sample types; (i) known F. hepatica status sera from an experimental infection for the comparison of sensitivities and specificities, (ii) sera from pre- and post-flukicide-treated (albendazole, closantel, nitroxynil and triclabendazole) beef cattle to contrast the differences of seropositivity before and after treatment, and (iii) bulk tank milk samples from dairy herds sampled during high and low F. hepatica exposure periods for assessing seasonal variations with the four tests available. Samples were tested using ELISA kits supplied by four manufacturers (Ildana Biotech, IDEXX, Svanova, and Bio-X). Samples were analysed simultaneously and in duplicate. Results In the control population Ildana, IDEXX and Bio-X presented 100% sensitivity (Se) and specificity (Sp), Svanovir presented a Se of 59% and a Sp of 96%. In flukicide-treated beef cattle, kits highlighted decreasing antibody levels 90 days post-treatment in variable degrees. Finally, bulk milk showed a significant decrease in ELISA value between high and low fluke exposure periods with all tests studied. Conclusions Se and Sp found in the present study, confirm that Ildana, IDEXX and Bio-X are accurate for the detection of F. hepatica exposure in Irish cattle. Svanovir Se and Sp in this population, indicate that a larger study is necessary to confirm this test characteristic in Irish herds. In post-treatment use, Bio-X showed a consistent and significant decrease of ELISA value in all groups treated, denoting to be a reliable tool for assessing treatment effect at 90 days post-treatment. Finally, all tests showed to be a reliable tool for the F. hepatica monitoring of high and low exposure seasons, using bulk tank milk samples.
    • Comparison of four commercially available ELISA kits for diagnosis of Fasciola hepatica in Irish cattle

      Munita, Maria Pia; Rea, Rosemary; Martinez-Ibeas, Ana Maria; Byrne, Noel; Kennedy, Aideen; Sekiya, Mary; Mulcahy, Grace; Sayers, Riona; Department of Agriculture, Food and the Marine; Dairy Research Ireland; et al. (Springer Science and Business Media LLC, 2019-11-21)
      Background: Fasciola hepatica is a liver parasite of mammals and it results in poor welfare outcomes and economic losses in ruminants. While faecal egg count is the test most commonly used for diagnosis, it does not indicate presence of migrating immature stages. Serological techniques increase sensitivity at all stages of the liver fluke infection. The aim of this study was to compare four commercially available ELISA tests for the diagnosis of F. hepatica. For this purpose, we tested three sample types; (i) known F. hepatica status sera from an experimental infection for the comparison of sensitivities and specificities, (ii) sera from pre- and post-flukicide-treated (albendazole, closantel, nitroxynil and triclabendazole) beef cattle to contrast the differences of seropositivity before and after treatment, and (iii) bulk tank milk samples from dairy herds sampled during high and low F. hepatica exposure periods for assessing seasonal variations with the four tests available. Samples were tested using ELISA kits supplied by four manufacturers (Ildana Biotech, IDEXX, Svanova, and Bio-X). Samples were analysed simultaneously and in duplicate. Results: In the control population Ildana, IDEXX and Bio-X presented 100% sensitivity (Se) and specificity (Sp), Svanovir presented a Se of 59% and a Sp of 96%. In flukicide-treated beef cattle, kits highlighted decreasing antibody levels 90 days post-treatment in variable degrees. Finally, bulk milk showed a significant decrease in ELISA value between high and low fluke exposure periods with all tests studied. Conclusions: Se and Sp found in the present study, confirm that Ildana, IDEXX and Bio-X are accurate for the detection of F. hepatica exposure in Irish cattle. Svanovir Se and Sp in this population, indicate that a larger study is necessary to confirm this test characteristic in Irish herds. In post-treatment use, Bio-X showed a consistent and significant decrease of ELISA value in all groups treated, denoting to be a reliable tool for assessing treatment effect at 90 days post-treatment. Finally, all tests showed to be a reliable tool for the F. hepatica monitoring of high and low exposure seasons, using bulk tank milk samples.
    • Liver fluke in Irish sheep: prevalence and associations with management practices and co-infection with rumen fluke

      Munita, Maria Pia; Rea, Rosemary; Martinez-Ibeas, Ana Maria; Byrne, Noel; McGrath, Guy; Munita-Corbalan, Luis Enrique; Sekiya, Mary; Mulcahy, Grace; Sayers, Ríona G.; Department of Agriculture, Food and the Marine; et al. (Springer Science and Business Media LLC, 2019-11-06)
      Background: The present study aimed to identify the national prevalence of Fasciola hepatica in Irish sheep and to conduct a risk analysis assessment based on management and treatment practices in participating focks. Also, co-infection with rumen fuke was quantifed and its association with liver fuke and management practices was assessed. Methods: A total of 305 sheep focks were selected ensuring even national representation of the sheep population. Participating farms were asked to complete a survey questionnaire on farm management practices and submit faecal samples during the winter of 2014–2015. Pooled faecal samples were analysed for the presence of F. hepatica and coinfection with rumen fuke. Apparent and true prevalence were calculated, additionally, the rate of co-infection with rumen fuke was also obtained. Correlation and regression analyses were used for assessing associations between management practices, liver fuke infection and co-infection with rumen fuke. Results: The national true prevalence of F. hepatica was 50.4% (n=305). Regional prevalence varied from 41% in the east to 52% in the south. Co-infection with rumen fuke was observed in 40% of the studied population and corre‑ lated with increased F. hepatica egg counts (OR=2.9; P≤0.001). Predominant breeds were Sufolk, Texel and Horned Mountain breeds. Beef cattle were the most frequent type of other livestock present on farms and mixed species grazing was frequently reported (73%). More than half of the focks reported a mid-to-late lambing period (MarchApril). Use of mountain land for grazing was of 32%. Flukicides were most commonly used twice over the autumnwinter period. Regression analyses highlighted signifcant association of F. hepatica status, with the presence of other livestock on farm, frequency of fukicides used during the winter and clinical presentation of liver fuke. A signifcant increase in eggs per gram of faeces was observed in Charollais sheep in comparison with all other breeds. Co-infec‑ tion with F. hepatica and Calicophoron daubneyi was also signifcantly associated with the presence of other livestock on the farm, type of fukicide used and clinical fasciolosis. Conclusions: The present study provides up-to-date information on the prevalence of F. hepatica in Irish sheep and adds insight to the epidemiology of the disease. These fndings will be useful for designing new holistic control meas‑ ures for F. hepatica infection.
    • Liver fluke in Irish sheep: prevalence and associations with management practices and co-infection with rumen fluke.

      Munita, Maria Pia; Rea, Rosemary; Martinez-Ibeas, Ana Maria; Byrne, Noel; McGrath, Guy; Munita-Corbalan, Luis Enrique; Sekiya, Mary; Mulcahy, Grace; Sayers, Riona; Department of Agriculture, Food and the Marine (Biomed Central, 2019-11-06)
      Background The present study aimed to identify the national prevalence of Fasciola hepatica in Irish sheep and to conduct a risk analysis assessment based on management and treatment practices in participating flocks. Also, co-infection with rumen fluke was quantified and its association with liver fluke and management practices was assessed. Methods A total of 305 sheep flocks were selected ensuring even national representation of the sheep population. Participating farms were asked to complete a survey questionnaire on farm management practices and submit faecal samples during the winter of 2014–2015. Pooled faecal samples were analysed for the presence of F. hepatica and co-infection with rumen fluke. Apparent and true prevalence were calculated, additionally, the rate of co-infection with rumen fluke was also obtained. Correlation and regression analyses were used for assessing associations between management practices, liver fluke infection and co-infection with rumen fluke. Results The national true prevalence of F. hepatica was 50.4% (n = 305). Regional prevalence varied from 41% in the east to 52% in the south. Co-infection with rumen fluke was observed in 40% of the studied population and correlated with increased F. hepatica egg counts (OR = 2.9; P ≤ 0.001). Predominant breeds were Suffolk, Texel and Horned Mountain breeds. Beef cattle were the most frequent type of other livestock present on farms and mixed species grazing was frequently reported (73%). More than half of the flocks reported a mid-to-late lambing period (March-April). Use of mountain land for grazing was of 32%. Flukicides were most commonly used twice over the autumn-winter period. Regression analyses highlighted significant association of F. hepatica status, with the presence of other livestock on farm, frequency of flukicides used during the winter and clinical presentation of liver fluke. A significant increase in eggs per gram of faeces was observed in Charollais sheep in comparison with all other breeds. Co-infection with F. hepatica and Calicophoron daubneyi was also significantly associated with the presence of other livestock on the farm, type of flukicide used and clinical fasciolosis. Conclusions The present study provides up-to-date information on the prevalence of F. hepatica in Irish sheep and adds insight to the epidemiology of the disease. These findings will be useful for designing new holistic control measures for F. hepatica infection.
    • Six-year longitudinal study of Fasciola hepatica bulk milk antibody ELISA in the dairy dense region of the Republic Ireland

      Munita, Maria P; Rea, Rosemary; Bloemhoff, Yris; Byrne, Nicky; Martinez-Ibeas, Ana M; Sayers, Riona; Irish Dairy Levy (Elsevier, 2016-09-28)
      Completion of the F. hepatica lifecycle is dependent on suitable climatic conditions for development of immature stages of the parasite, and its snail intermediate host. Few investigations have been conducted regarding temporal variations in F. hepatica status in Irish dairy herds. The current study aimed to conduct a longitudinal study examining annual and seasonal trends in bulk milk seropositivity over six years, while also investigating associations with soil temperature, rainfall and flukicide treatment. Monthly bulk milk samples (BTM) were submitted by 28 herds between March 2009 and December 2014. In all, 1337 samples were analysed using a Cathepsin L1 ELISA. Soil temperature, rainfall and management data were obtained for general estimating equation and regression analyses. A general decrease in milk seropositivity was observed over the six year study period and was associated with an increased likelihood of treating for liver fluke (OR range = 2.73–6.96). Annual and seasonal analyses of rainfall and F. hepatica BTM status yielded conflicting results. Higher annual rainfall (>1150 mm) yielded a lower likelihood of being BTM positive than annual rainfall of <1000 mm (OR = 0.47; P = 0.036). This was most likely due to farmers being more proactive in treating for F. hepatica in wetter years, although a ‘wash effect’ by high rainfall of the free living stages and snails cannot be ruled out. Higher seasonal rainfall (>120 mm), however, was associated with increased ELISA S/P% values (Coefficient = 9.63S/P%; P = 0.001). Soil temperature was not found to influence F. hepatica to the same extent as rainfall and may reflect the lack of severe temperature fluctuations in Ireland. Flukicides active against both immature and mature F. hepatica were approximately half as likely to record a positive F. hepatica herd BTM status than a flukicide active against only the mature stage of the parasite (OR ≅ 0.45; P < 0.01). This study highlights the importance of examining both annual and seasonal F. hepatica data, which can vary significantly. Additionally, it highlights the progress that can be achieved in fluke control by application of a continuous BTM monitoring program.