Prevalence of flukes (Fasciola hepatica and paramphistomids) in cattle in south-eastern Mexico.

The objective of this study was to determine the risk factors and prevalence of trematodes in south-eastern Mexico. The prevalence of trematodes was determined in 1010 bovines. The study was carried out from October 2018 (n=291) to December 2019 (n=719). Only in 2019 rumen and liver fluke eggs were differentiated. Faecal samples (n=311) were obtained from farms in southeast Mexico located in Tabasco, Chiapas and Campeche. In addition, the presence of flukes in liver and rumen from slaughtered cattle in abattoirs was recorded with a total of 408 samples. A logistic procedure was used to obtain the prevalence and the effect of main risk factors such as land physiography (flooded areas and hills), year, sex, animals' age and type of sample obtained (eggs in faeces and flukes). The general prevalence of flukes in cattle was 32.3 % in 2018 and 41.7 % in 2019. Prevalence of F. hepatica (liver fluke) was 18.6 % (134/719) and that of paramphistomids (rumen fluke) was 33.4 % (240/719). The infected cattle from the slaughterhouse indicated a lower prevalence of F. hepatica (1 %) and rumen fluke (26.7 %) than in farms detected by egg in faeces (41.8 % and 42.1 %, respectively). The physiographic zone was decisive in the presence of F. hepatica and rumen fluke, while sex did not represent a risk factor (P > 0.05). The environmental conditions of the Mexican southeast favour the presence of both liver and rumen fluke.

Soil fungi enable the control of gastrointestinal nematodes in wild bovidae captive in a zoological park: a 4-year trial.

The control of gastrointestinal nematodes among ruminants maintained in zoological parks remains difficult due to infective stages develop in the soil. For the purpose to improve the possibilities of the control of gastrointestinal nematodes (genera Trichostrongylus, Nematodirus, Chabertia and Haemonchus) affecting wild captive bovidae ruminants belonging to the subfamilies Antilopinae, Caprinae, Bovinae and Reduncinae, commercial pelleted feed enriched with a blend of 104-105 spores of both filamentous fungi Mucor circinelloides + Duddingtonia flagrans per kg meal was provided for a period of 3.5 years. All animals were dewormed at the beginning of the trial and also when exceeding a cut-off point of 300 eggs per gram of feces (EPG). The anthelmintic efficacy ranged between 96 and 100%. The need for repeating the administration of parasiticide treatment disappeared at the 24th month of study in the Antilopinae individuals, and at the 8th month in the Caprinae, Bovinae and Reduncinae. No side-effects were observed on the skin or in the digestive, respiratory or reproductive system. It was concluded that this strategy provides a sustainable tool for preventing the contamination of paddocks where captive ruminants are maintained, decreasing the risk of infection by gastrointestinal nematodes and consequently the need of frequent deworming.

Antigen characterization from second instars of oestrid bot flies for the detection of anti-Cephenemyia stimulator antibodies by ELISA in roe deer (Capreolus capreolus).

A study to determine the most appropriate antigen for use in the serodiagnosis of Cephenemyia (Diptera: Oestridae) infestation in roe deer (Capreolus capreolus) was carried out using immunoenzymatic tests. Serum samples from 43 roe deer from northern Spain were obtained post-mortem and corresponding numbers of bot fly larvae established. Three antigen complexes were tested, including Cephenemyia stimulator Clark excretory/secretory antigens (CsES), C. stimulator somatic antigens (CsSA) and Oestrus ovis L. (Diptera: Oestridae) excretory/secretory antigens (OoES). In addition, the composition of each antigen was analysed using an electrophoresis system. Cephenemyia stimulator larvae were found in 25% of roe deer; the mean intensity of infection was 24.3 larvae per infested animal. In the antigen analysis, CsSA showed four exclusive bands of molecular weight (17-19, 62, 65 and 67-70 kDa). A positive correlation between immunoglobulin G (IgG) values and total number of larvae was found with CsES and CsSA. The highest sensitivity value, negative predictive value and negative likelihood ratio were obtained using CsES. The highest specificity value, positive likelihood ratio and kappa value were achieved with CsSA. The predictive values of the enzyme-linked immunosorbent assay (ELISA) using CsES and CsSA reached statistical significance and seroprevalence values were 26-44%. The use of ELISA with CsES and CsSA seems promising in the non-invasive diagnosis of Cephenemyia infestation in roe deer.

Application of the analysis of serum antibodies (immunoglobulins M and G) to estimate the seroprevalence of ovine oestrosis and to evaluate the effect of chemotherapy.

The analysis of immunoglobulin (Ig) M and IgG serum isotypes as indicators of the presence of Oestrus ovis developmental stages was carried out. A serological survey to discover the seasonal variations in the prevalence of oestrosis in sheep from an oceanic climate area was developed. Six hundred and sixty-nine blood samples were analyzed for the presence of IgM and IgG using an enzyme-linked immunosorbent assay and O. ovis second-stage larvae excretory/secretory antigens. In addition, the effect of an ivermectin-based treatment on the humoral immune response was measured. The percentage of positive animals was 54% for IgM and 55% for IgG. The highest percentages of sheep positive to IgM were recorded in the winter and to IgG in the summer. No animals positive to IgM were detected in the ivermectin-treated sheep. These results seem to underline that oestrosis is a parasitic disease to keep watch for in regions with an oceanic climate. A noninvasive evaluation of the success of chemotherapy seems possible by analyzing the serum IgM response.

The efficacy of predatory fungi on the control of gastrointestinal parasites in domestic and wild animals-A systematic review.

BACKGROUND: Gastrointestinal parasites like nematodes are associated with significant impacts on animal health, causing poor growth rates, diseases and even death. Traditional parasite control includes the use of anthelmintic drugs, albeit being associated with drug resistance and ecotoxicity. In the last decade, biological control of parasites using nematophagous or predatory fungi has been increasingly studied, although systematic evidence of its efficacy is still lacking. The aim of this work was to assess the evidence of efficacy of nematophagous fungi in the control of nematodes and other gastrointestinal parasites in different animal species. METHODS: Using the PICO method (Population, Intervention, Comparison and Outcomes), we performed a systematic review on the subject to search for original papers published between January 2006 and October 2019, written in English, and indexed in PubMed/Medline. Medical Subject Headings (MeSH) terms were used in the syntax. Papers were selected for detailed review based on title and abstract. Inclusion and exclusion criteria were applied, and relevant data were collected from the remaining papers. RESULTS: The literature search retrieved 616 papers. Eighty-nine were submitted to a detailed review. In the end, 53 papers were included in the analysis. The studies were very heterogeneous, using different fungi, doses, frequency of administration, duration of treatment, host animals, and target parasites. Considering the 53 papers, 44 studies (83 % of the interventions) showed efficacy, with only 9 studies (17 %) showing no significant differences when compared to control. CONCLUSION: With the increasing hazards of drug resistance and ecotoxicity, biological control with predatory fungi stands out as a good tool for future parasite management, whether as a complementary treatment or as an alternative to standard parasite control.

Horses are susceptible to natural, but resistant to experimental, infection with the liver fluke, Fasciola hepatica.

Fasciola hepatica is a common parasite of livestock in Ireland, causing significant economic losses and affecting animal welfare. A previous abattoir study of 200 horses led to an estimated 9.5 % prevalence of infection in horses slaughtered in Ireland. However, the epidemiology and pathogenic significance of this infection in this species is not well-described. The objectives of this study were to determine the susceptibility of horses to oral challenge infection with F. hepatica metacercariae, and to document the course of the infection along with serological and biochemical response. We attempted an experimental infection of horses (n = 10; 9 geldings and 1 mare) with F. hepatica. Four were given 1000 metacercariae, four 500 metacercariae and two were sham-infected. Blood and faecal samples were taken at intervals up to 18 weeks post-infection (wpi). ELISA assays were used to assess sero-conversion in the experimental horses and also in a panel of sera from horses of known fluke status. No flukes were recovered from any of the livers, and neither were any lesions that could be attributed to F. hepatica infection observed. Coproantigen ELISA was negative throughout for all horses. Three antibody detection ELISAs, useful in diagnosing fasciolosis in other species, had limitations as diagnostic aids as determined using a panel of sera from horses of known F. hepatica infection status. This study is limited by the relatively small number of animals included, and the relatively short duration of the study period. Failure to establish infection after oral challenge raises fundamental questions on the pathophysiology and epidemiology of equine fasciolosis.

Clinical trial of efficacy of ivermectin pour-on against gastrointestinal parasitic nematodes in silvopasturing horses.

The aim of this study was to assess, by a clinical trial, the efficacy of an ivermectin-based pour-on treatment against gastrointestinal parasitic nematodes in naturally infected horses using 2 groups of mature indigenous Pura Raza Galega grazing mares. Faecal and blood samples were collected individually over a 21 week period. Faeces were analysed by the coprological flotation, sedimentation and migration techniques. Changes in circulating blood cells were monitored over the study period. The administration of the ivermectin suppressed the egg-elimination of ascarids and pinworms throughout the study and no strongyle-eggs were observed in the treatment group between the 3rd and 10th weeks. The numbers of red cells increased significantly after the anthelmintic therapy, and a statistical reduction in circulating leucocytes was recorded. No side effects were observed. The pour-on ivermectin formulation was highly successful against gastrointestinal nematodes and appears to be a useful therapeutic routine for large groups of horses.

Field evaluation for anthelmintic-resistant ovine gastrointestinal nematodes by in vitro and in vivo assays.

A coprological survey to analyze the presence of flock resistance to benzimidazoles (BZ) and macrocyclic lactones (ML) was performed in sheep under field conditions. Fecal samples were collected from 2,625 sheep in 72 commercial farms from Galicia (NW Spain). The in vitro (FECRT, fecal egg count reduction test) and in vivo (EHA, egg hatch assay, and LFIA, larval feeding inhibition assay) tests were used to assess the efficacy of these anthelmintics. Coprocultures were also developed to obtain knowledge on the main genera of trichostrongylid nematoda prior to, and after, the administration of the anthelmintics. By using the FECRT, BZ resistance was observed in 13 (18%) flocks, whereas ML resistance was only detected in 2 (3%) farms. The number of resistant flocks to BZ was 21 (29%) by using the EHA and 7 (10%) by means of the LFIA. None of the flocks used in this study showed simultaneous resistance to both employed anthelmintics. The results from the in vitro and in vivo tests revealed that 92% of the flocks FECRT resistant to BZ were also resistant with the EHA. The LFIA confirmed all the farms resistant to ML by using the in vivo test. After the administration of BZ, nematode larvae belonging to Teladorsagia circumcincta (32.2%), Trichostrongylus spp. (29%), Nematodirus spp. (6.5%), and Chabertia ovis (3.2%) were identified. In the flocks receiving ML, only T. circumcincta was identified (57%). We recommend the use of in vitro tests because they are more efficient. As the use of macrocyclic lactones is increasing in this region, further investigation is needed for detecting resistance to the anthelmintic family compounds by the LFIA.

Assessment of climatic and orographic conditions on the infection by Calicophoron daubneyi and Dicrocoelium dendriticum in grazing beef cattle (NW Spain).

From February 2003 to March 2004, 1148 faecal samples from autochthonous Rubia Gallega cattle breed were collected in 170 farms from Lugo (NW Spain), in order to find out the prevalence and intensity of egg-excretion by Calicophoron daubneyi and Dicrocoelium dendriticum, and the possible influence of external factors as annual mean temperature and rainfall, altitude and mean slope. Twenty-six percent (95% CI 19%, 33%) and the 18% (12, 24) of the farms were infected with rumen and lancet flukes. C. daubneyi egg-output (Me=24) was found in 13% (11, 15) of the individual samples. Six percent (5, 7) of the animals in the study had low D. dendriticum eggs (Me=8). By estimating the odds ratio values it was shown that the highest probability of infection by C. daubneyi was in pastures with a mean slope of less than 13% (OR=1.9) and situated under 600 m (OR=1.6). Annual mean rainfall and temperature were not identified as risk factors for the infection with rumen flukes. Mountainous pastures with a mean slope of higher than 25% (OR=5.8) and situated over 600 m (OR=24.6) where precipitation was high (>1000 mm; OR=7) and temperature low (<11 degrees C; 2.8) had involved the highest risk of infection by D. dendriticum. Because of the prevalences found in this study, employment of suitable management practices with strategic treatments with efficacious anthelmintics are needed to reduce the presence of both trematodes and to increase the health status of grazing beef cattle.

Immunodiagnosis of current fasciolosis in sheep naturally exposed to Fasciola hepatica by using a 2.9 kDa recombinant protein.

A 2.9 kDa recombinant-Fasciola hepatica protein (FhrAPS) was employed to estimate the prevalence of fasciolosis in sheep maintained under field conditions. For this purpose, 340 samples with known status in relation to fasciolosis by using a direct-ELISA and the coprological sedimentation were used. These samples were analysed by using an indirect-ELISA (iELISA) and the FhrAPS recombinant protein and excretory/secretory antigens (FhES) of this trematode. Current fasciolosis (CF) was named when results were positive to antigenemia and/or coprology. Out of 198 sheep with current fasciolosis, 68% were positive to the FhrAPS-ELISA test and 53% to the FhES. We observed 14% of the CF-neg sheep were positive to the FhrAPS, whereas this percentage was 52% with the FhES. A significant correlation between FhrAPS and current fasciolosis was obtained (r2=0.513, p=0.001). We concluded that the FhrAPS provides a more suitable antigen than FhES for developing field trials to know the prevalence of early and current fasciolosis.

Analysis of the IgG antibody response against Paramphistomidae trematoda in naturally infected cattle. Application to serological surveys.

The IgG antibody response to Calicophoron daubneyi (Digenea: Paramphistomidae) excretory/secretory antigens was evaluated in naturally infected cattle from Lugo (Galicia, NW Spain) by using an ELISA procedure. Two studies were conducted, first a survey in 524 cattle separated into three groups according to age, G-1 (0-2 years old), G-2 (3-5 years old) and G-3 (> 6 years old). In the second study, three groups of cattle were employed: G-I, naturally infected; G-T, naturally infected and treated with oxyclozanide plus levamisole (Nilzan Plus); G-C, cattle maintained in a farm where C. daubneyi has never diagnosed. Variations on egg-output and haematic parameters (erythrocytes, haematocrite, leukocytes and lymphocytes) were also analyzed. The ELISA procedure showed that 61.2% of the cattle in the first study had been exposed to the trematode, but only 10.1% passed eggs in the feces. Age-association with egg-output was shown but not with the IgG values. In the second experiment, the administration of the anthelmintic reduced significantly the IgG kinetic levels and the C. daubneyi-egg-output was suppressed during 12 weeks in the G-T group. The values of red cells, haematocrite, leukocytes and lymphocytes increased significantly in the treated cattle 5 weeks after chemotherapy; however, new reduction after week 5 was recorded, as results of the challenge of these cattle. This is the first investigation in which evaluation of the IgG humoral response against C. daubneyi in cattle has been carried out. We proved that a notable IgG response in naturally infected cattle is induced, and can be detected by using an ELISA procedure. The IgG antibodies did not increase after challenge infection. Our results proved an important percentage of cattle were exposed to this trematode in the area of study and suitable measures for preventing this relationship must be considered.

A 2.9 kDa Fasciola hepatica-recombinant protein based ELISA test for the detection of current-ovine fasciolosis trickle infected.

The suitability of an enzyme linked immunosorbent assay (ELISA) test with a 2.9 kDa Fasciola hepatica-recombinant protein (FhrAPS) for diagnosing early and current-ovine fasciolosis was analyzed, and compared to that obtained by using a direct ELISA for detecting F. hepatica-circulating FhES antigens and to the coprological sedimentation for fluke egg quantitation. Fourteen Gallega autochthonous breed sheep were experimentally infected with metacercariae by a trickle system (small repetitive infections) and divided into two groups: G-I represented a primary infection for 34 weeks; G-R, animals with primary infection and reinfected 18 w.a.p.i. Seven sheep were left uninfected as the control group (G-C). Serum IgG antibody values against the FhrAPS rose rapidly by 1st w.a.p.i. in all infected sheep. Antibody levels in those with primary infection (G-I, G-C) peaked at 10 weeks, diminishing slightly and levelling from 16 to 34 weeks. Those with primary infection reinfected at 18 weeks had a rebound effect with the highest values observed. Circulating F. hepatica-ES antigens were detected by the 1st w.a.p.i. in all infected groups peaking at 6 weeks, decreasing rapidly to uninfected control values by 10 weeks of infection. Faecal egg-output started 11 weeks after primary infection. An increase in the IgG antibody as well as antigen responses to the FhrAPS and to anti-FhES from the 18 w.a.p.i. was recorded in G-T and G-R after the challenge infection. Antibody levels remained high whereas antigenemia values diminished after 6 weeks. A positive significant correlation between the IgG response against the FhrAPS and the F. hepatica circulating antigens (r2 = 0.428, p = 0.001) was obtained. In conclusion, our standardized diagnostic ELISA for fasciolosis based on the detection of IgG responses to the FhrAPS would be a valuable tool to diagnosis early and current F. hepatica-infections in sheep.

Prevalences of gastrointestinal parasites in sheep and parasite-control practices in NW Spain.

A coprological survey to know the presence of gastro-intestinal nematode parasites infecting sheep was done in Galicia (NW Spain), an area with Atlantic climate where sheep production is replacing cattle due to the Agricultural Community Politics of the European Union. From September 2001 to November 2002, 1710 faecal samples were randomly collected from 49 sheep farms and examined by using the flotation technique to determine the prevalence of gastro-intestinal nematode parasites. The sheep-level prevalence was 100%, and the genera identified were Chabertia, Cooperia, Haemonchus, Nematodirus, Oesophagostomum, Teladorsagia, Trichostrongylus and Trichuris spp. A questionnaire was distributed to the farmers (at the same time as sampling) about parasite-control practices during the year before sampling (2000). Ninety percent (95% CI 81%, 98%) of the farmers said they used antiparasitic drugs occasionally, but none of them asked for a coprological analysis prior to the treatment and the efficacy of the drugs was never evaluated. A higher median EPG was observed in the treated sheep (163) than in the untreated ones (26). Chemotherapy was the only parasite-control practice. Flocks that treated according to the farmer's previous experience had higher median EPG (236) than other flocks (185 following the prescription of their veterinary clinician and 232 based on the commercial agent's counsel). Typically, one benzimidazole-treatment per year was applied in autumn and non-veterinary counsel was used. Sheep treated with imidotiazoles had lower EPG (144) than other drugs (164 for the benzimidazoles and 166 for the macrocyclic lactones). We found a higher median EPG in the sheep receiving two treatments/year (175) than in those treated only once per year (156). These results suggest lack of knowledge about worm-control strategies and anthelmintic use or unwillingness to apply such knowledge.

Analysis of the humoral immune response to Oestrus ovis in ovine.

Antibody responses (IgG, IgM and IgA) against Oestrus ovis were analyzed in sheep and in first year grazing lambs from Sardinia (Italy) by an indirect-enzyme-linked immunoassay test and L2 O. ovis excretory/secretory antigens. Serum samples from 208 sheep were obtained prior to be slaughtered, and then heads were removed and cut open along their longitudinal axis to collect the parasites from the nasal cavities, turbinates and sinus. Besides this, blood samples were monthly collected from the lambs of G-1 (maintained under field conditions) and the lambs of G-2 (kept housed since birth to avoid Oestrus infestations) throughout a year. In the sheep, a positive significant correlation was observed between the number of first instar O. ovis larvae and the values of IgM, and between the second instar larvae and the IgG optical densities. In the lambs, all classes of antibodies increased significantly from July in G-1. The highest values of IgG were reached in September (IgG) and decreased in November-December. The IgM response peaked in November, and very low values of IgA were observed during the study. Matching these data with chronobiology of O. ovis in this region, we conclude that the first infection occurs on May, stimulating the production of humoral antibodies. The reduction of the IgG antibody levels starting from October means the beginning of the diapause while the IgM response seems to be associated to the presence of L1 in the nasal cavities. The data obtained led us to forecast an early treatment of the ovine on June-July, which should keep away from the maturation of O. ovis L1 larvae, avoiding the development of clinical lesions and interrupting the life cycle of this parasite.

Chronobiology of Oestrus ovis (Diptera: Oestridae) in Sardinia, Italy: guidelines to chemoprophylaxis.

Oestrus ovis (Linné 1761) larvae are obligatory parasites of the nasal and sinus cavities of sheep and goats. Infestation is prevalent in hot and dry regions, such as Mediterranean countries. The current work was developed to establish the chronobiology of O. ovis in Sardinia, to determine the most suitable time for chemoprophylaxis. A survey was carried out during 1998, and sheep heads were collected monthly from local flocks. A total of 443 heads was examined, and the prevalence of oestrosis was 73.8%. We collected 2,691 larvae (mean = 6.07 +/- 9.52), and the intensity was greatest in November. The humoral immune response against the nasal bot fly was analyzed by means of an indirect-ELISA using second-instar O. ovis excretory and secretory antigens. A seasonal variation in the antibody levels was observed, increasing from April and peaked in June and in September. A significant correlation was observed between first instar intensity and the mean relative humidity (r2 = 0.120; P < 0.05), and between second-instar intensity and the mean temperature (r2 = 0.241; P < 0.05). Three periods in the chronobiology of O. ovis were defined: diapause (October-February), the active phase ofthe endogenous cycle (March-September) and the exit phase (May-September). Our results showed that treatment in October-November was suitable, because first instars were in diapause, preventing the development of first into second instars, and second into third instars.

Use of a sandwich-enzyme-linked immunosorbent assay (SEA) for the diagnosis of natural Fasciola hepatica infection in cattle from Galicia (NW Spain).

An indirect-enzyme immune-linked immunosorbent assay (IEA) was compared to a sandwich-enzyme-linked immunosorbent assay (SEA) to diagnose fasciolosis in naturally infected cattle. By means of a rabbit polyclonal IgG antibody to F. hepatica antigens, captured circulating antigens were detected by an SEA technique. A total of 85.1% of the samples examined was considered positive by using the IEA-test, whereas this percentage was of 37.3% by the SEA test. Using the two enzymatic probes, only the 38.3% resulted positive to both immunoassay probes. Our data indicate that circulating antigen detection assay in combination with results of antibody detection may offer a more reliable diagnostic technique than classic coprologic ones, with results very appropriate for epidemiologic studies carried out on a wide area.

Effect of treatment on the dynamics of circulating hypodermin C in cattle naturally infested with Hypoderma lineatum (Diptera: Oestridae).

An antigen capture ELISA, using a murine monoclonal antibody recognising recombinant hypodermin C (rHyC), was used to evaluate the influence of early treatment with eprinomectin (Eprinex) or fenthion (Spotton) on the kinetics of circulating hypodermin C in calves naturally infested with Hypoderma lineatum. No viable larvae were collected from treated animals, whereas a variable number of warbles were found in control animals. Treatment provoked a decrease in circulating HyC levels that was significant 9 days post-treatment (p.t.). Circulating antigen levels in the treated cattle remained detectable for approximately 99 days p.t. In contrast, control animals had no detectable antigen at 64 days p.t., 42 days earlier than in the treated animals. These results suggest that larvae were either gradually killed, resulting in slow release of antigen or they were encapsulated, leading to the slow liberation of antigen. Kinetics of circulating HyC did not differ among the two insecticide treatments. Antibodies persisted, in all groups, throughout the 120-day study. These results suggest that the antigen capture ELISA will be useful as a technique for detecting successful treatment of cattle grub infestations and for the detection of new infestations in previously infested cattle.

Serum antibodies to Dicrocoelium dendriticum in sheep from Sardinia (Italy).

An indirect enzyme-linked immunosorbent assay (ELISA) with Dicrocoelium dendriticum excretory/secretory antigens was used to evaluate the presence of serum antibodies against the trematode in 738 sheep randomly chosen in Sardinia (Italy). Coprological sedimentation was used to discover egg-output. Seropositivity was detected in 86.2% tested sheep, whereas faecal prevalence was 6.7%; all that were faecal-positive also were ELISA-positive.

Influence of age and breed on natural bovine fasciolosis in an endemic area (Galicia, NW Spain).

An analysis was undertaken of the effect of breed and age on bovine fasciolosis using antibody and antigen detection ELISAs. A total of 84.3% of the 1284 serum samples examined had positive antibody values and 20.4% exhibited antigenaemia. The seroprevalence of antibodies in crossbred cattle was higher than that in autochthonous Rubia Gallega, Friesian or Brown Swiss cows. The highest percentage antigenaemia occurred in the Brown Swiss cattle, but the breed differences were not statistically significant. Cattle aged 3-5 years had the highest antibody and circulating antigen prevalence and the age differences were significant. It was concluded that the apparent influence of breed was probably closely associated with the husbandry system. The autochthonous Rubia Gallega may be better adapted to fasciolosis as its percentage of antigenaemia was the lowest.

Relationship between exposure to Fasciola hepatica in roe deer (Capreolus capreolus) and cattle extensively reared in an endemic area.

The aim of this work is to know the prevalence of Fasciola hepatica in 301 roe deer and in 676 beef cattle kept in an endemic area. Detection of antibodies was determined in roe deer using a homemade ELISA with excretory/secretory antigens (FhES) and a recombinant protein (FhrAPS). None of the deer passed eggs by faeces and none flukes in their livers were found. The seroprevalence of F. hepatica was 29% using FhES, with significantly higher values in the oldest ones (36%). Twenty-eight percent of the samples were positive to FhrAPS. Twenty-three percent of the cows eliminated eggs of F. hepatica and the seroprevalence was 67% using FhrAPS. No relationship between the seropositivity values of deer and cattle was demonstrated. The role of wild ruminants as reservoirs of F. hepatica is discussed. We encourage the use of ELISA to know the possibility of exposure to trematodes in wild ruminants.