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PubMed2005

Performance characteristics of an enzyme-linked immunosorbent assay performed in milk for the detection of liver fluke (Fasciola hepatica) infection in cattle.

AIM: To determine the performance characteristics of an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against liver fluke (Fasciola hepatica) in bovine milk. METHODS: Serum and milk from liver fluke infected and non-infected cattle was assayed in a commercially available enzyme-linked immunosorbent assay. Serum test results were used to determine the "gold standard" infection status of cattle and milk ELISA results assessed by ROC analysis. RESULTS: ROC analysis suggested changes to the ELISA protocol, arriving at milk dilutions assayed considerably higher than those suggested by the manufacturer. With those changes, the ELISA performed with high sensitivity and specificity, 95 and 98.2%, respectively, for individual bovine milks (relative to sera). For bovine tank milks, sensitivity was lower, with bulk milks only testing positive if 60% or more of cattle milking in the herd were infected. CONCLUSIONS: The analysis of the ELISA's performance when used on individual bovine milks demonstrated high sensitivity and specificity. ROC analyses optimised the assay conditions and cut-off point suggested by the manufacturer for this commercial diagnostic assay. This would help with the identification and control of fasciolosis, enabling simpler sample collection.

Animals↗
PubMed1999

Kinetics of antibody-based antigen detection in serum and faeces of sheep experimentally infected with Fasciola hepatica.

The monoclonal antibody ES78 was used in a sandwich immunosorbent assay (Sandwich ELISA) for the detection of antigens in sera and faeces in the course of Fasciola hepatica infection in 10 experimentally infected sheep. All infected sheep had circulating antigens in the first week post-infection (WPI). Antigenemia was detectable until WPI 3 in four infected sheep, WPI 4 in five infected sheep and in only one sheep by WPI 5. The detection of coproantigens (Fag) was possible in five infected sheep at WPI-4, in four sheep at WPI-5 and in one sheep only at WPI-6. This technique was compared to an indirect ELISA for the detection of antibodies using excretory secretory antigens of F. hepatica. A significant correlation was found between Fag and egg output and also with adult worm numbers. Our method demonstrated that the diagnosis of active fasciolosis in sheep is possible during all periods of infection.

Animals↗
PubMed1985

Efficacy and safety of clorsulon used concurrently with ivermectin for control of Fasciola hepatica in Florida beef cattle.

The safety and anthelmintic efficacy of clorsulon (7 mg/kg of body weight, orally) given concurrently with ivermectin (0.2 mg/kg, subcutaneously) for control of Fasciola hepatica were evaluated in 75 crossbred beef cattle. Twenty-three control animals were given only ivermectin. Fluke egg counts were significantly lower (P less than 0.0001) in clorsulon-treated animals by day 14 after treatment than before treatment (base line) and were significantly lower (P less than 0.005) after treatment in clorsulon-treated animals than in controls. There was no significant change in fluke egg counts of controls after treatment. The efficacy of clorsulon in reducing F hepatica egg counts was 99%. Clorsulon showed poor efficacy against paramphistomes. Adverse reactions were not observed.

Animals↗
PubMed1994

Fasciola hepatica: a study of the shedding of cercariae from Lymnaea truncatula raised under constant conditions of temperature and photoperiod.

Investigations on the shedding of cercariae of Fasciola hepatica were carried out in Lymnaea truncatula in order to verify the existence of a low-frequency periodicity in the numerical distribution of metacercariae per snail and per day when the snails are raised under controlled conditions. Preadult L. truncatula were thus collected in the field, individually exposed to two miracidia, and subsequently raised until their death under constant temperature (20 degrees C) and photoperiod (12 h/12 h diurnal rhythm). The 102 snails shedding parasites produced 24,325 metacercariae of which 5% were floating cysts. The daily production of cercariae was maximal during the first 30 days of the patent period, subsequently decreased until day 114, and ceased on day 124. No infradian-type rhythm was noted in the daily distribution of mean values. The snails shed their cercariae in one to 14 waves; 20.6% and 15.7% of the snails produced their parasites in four and five periods respectively. The authors suggest that the seven-day periodicity found by Audousset et al. (1989) in the daily distribution of cercariae produced by three colonies of L. truncatula raised in seminatural conditions must be attributed only to the influence of environmental factors.

Animals↗
PubMed2000

Kinetics of antibody-based antigen detection in serum and faeces of sheep experimentally infected with Fasciola hepatica.

The monoclonal antibody ES78 was used in a sandwich immunosorbent assay (Sandwich ELISA) for the detection of antigens in sera and faeces in the course of Fasciola hepatica infection in 10 experimentally infected sheep. All infected sheep had circulating antigens in the first week post-infection (WPI). Antigenemia was detectable until WPI 3 in four infected sheep, WPI 4 in five infected sheep and in only one sheep by WPI 5. The detection of coproantigens (Fa(g)) was possible in five infected sheep at WPI-4, in four sheep at WPI-5 and in one sheep only at WPI-6. This technique was compared to an indirect ELISA for the detection of antibodies using excretory secretory antigens of F. hepatica. A significant correlation was found between Fa(g) and egg output and also with adult worm numbers. Our method demonstrated that the diagnosis of active fasciolosis in sheep is possible during all periods of infection.

Animals↗
PubMed2006

Excretory-secretory product of fasciola hepatica worm protects against Schistosoma mansoni infection in mice.

The objective of this study was to evaluate the protective immunity of excretory-secretory products of Fasciola hepatica (FhES) worm against S.mansoni infection in mice. Evaluation of FhES antigen was through measuring worm burden, ova count, granuloma size and frequency as well as the histopathological picture of the liver. The study was extended to determine the level of free radical scavengers; lipid peroxide, glutathione (GSH), vitamin C, vitamin E, catalase and superoxide dismutase (SOD). Liver function enzymes such as aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were also taken into consideration. Four groups of eight mice each were selected for this study. Group 1 served as control group. Group 2: normal healthy mice vaccinated with FhES product. Group 3: S.mansoni infected mice for 2 months and group 4: infected mice pre-vaccinated with FhES antigen. Vaccination schedule comprised of a single subcutaneous injection of FhES antigen emulsified with Freund's complete adjuvant in a dose 0.5 mg protein/mouse, followed by intraperitoneal injections of the same antigen without adjuvant in 3 doses/week for 3 successive weeks. The total antigen inoculation was 5 mg protein/mouse. The present results revealed a drastic change in all the measured parameters after S.mansoni infection and a noticeable improved level after vaccination with FhES antigen. It can be concluded that FhES antigen succeeded to protect mice against schistosomiasis by a significant reduction in worm burden, ova count, granuloma size and number, improvement in the histopathological architecture of the liver as well as amelioration in the antioxidant levels under investigation.

Animals↗
PubMed1988

Flukicidal action of closantel against immature and mature Fasciola hepatica in experimentally infected rats and sheep.

The relative importance of peak level- and residual level-related flukicidal activity of closantel against immature and mature Fasciola hepatica was evaluated in a comparative efficacy trial using two animal species with a different plasma elimination pattern, that is, the rat and the sheep with an elimination half-life of less than one week and of two to three weeks, respectively. The rats were dosed orally with closantel at 20 mg kg-1 at two, four, six, eight and 10 weeks; the sheep at 10 mg kg-1 at eight, 10 and 12 weeks after artificial infection. Necropsy was performed either one week after treatment or 12 weeks after infection. Efficacy rates and the length of the recovered flukes were evaluated. It was demonstrated that the flukicidal effect of closantel is directly related to its peak plasma levels and less to its residual plasma concentrations. In the rat, a high efficacy (P less than 0.001) could be demonstrated against immature stages of four weeks or older. The two-week immature stages were less markedly affected. No significant differences in efficacy and size of the flukes were noted between the animals autopsied one week after treatment and those autopsied 12 weeks after infection. In the sheep, the efficacy against six-week and eight-week-old immature stages varied between 70.3 and 76.8 per cent and between 92.8 and 96.5 per cent, respectively. As in the rats, no marked differences in efficacy were noted between the animals autopsied one week after treatment and those autopsied 12 weeks after infection.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1996

Detection of Fasciola hepatica antigen in cattle faeces by a monoclonal antibody-based sandwich immunoassay.

A monoclonal antibody-based sandwich immunoassay (mAb sandwich ELISA) was developed for the detection of Fasciola hepatica antigen in the faeces of cattle. The assay was applied to samples from 100 cattle infected with F hepatica, 56 animals with parasitologically proven infections of other parasites and 100 uninfected animals. F hepatica antigen was detected in all the faecal samples from animals with fasciolosis, but none of the samples from the uninfected animals or from those with other parasitic infections had significant levels of F hepatica antigens. The results indicate that the mAb sandwich ELISA is a rapid, simple and useful method for the diagnosis of active F hepatica infection in cattle.

Animals↗
PubMed1981

The heterologous protection of rats against a challenge with Fasciola hepatica by prior infection with the nematode Nippostrongylus brasiliensis.

Previous infection of rats with nippostrongylus brasiliensis was shown to result in protection against an oral challenge with Fasciola hepatica metacercariae but not against an intraperitoneal challenge with newly excysted juvenile (NEJ) flukes. The timing of the challenge was important and a double infection with the nematode gave more consistent results than a single. Resistance appeared to be associated with a prior induction of intestinal eosinophilia. Sera from these resistant rats, however, failed to induce eosinophil adherence to NEJ flukes in vitro.

Animals↗
PubMed1983

Fasciola hepatica: increase of glycogen phosphorylase activity due to prostaglandins.

Both prostaglandin E1 (PGE1) and prostaglandin F2 alpha (PGF2 alpha) stimulate the glycogen phosphorylase (EC 2.4.1.1.) activity of Fasciola hepatica. Whole or sliced parasites were incubated with PGE1 (2.8 X 10(-7) and 2.8 X 10(-5) M) and PGF2 alpha (2.1 X 10(-7) and 2.1 X 10(-5) M) and enzyme activity was measured in homogenates prepared immediately following the incubation. No substantially different effect was noted between the two assayed doses of prostaglandins. Prostaglandins appeared to be less effective in sliced parasites.

Alprostadil↗
PubMed2005

Fasciola hepatica: epidemiological surveillance of natural watercress beds in central France.

A total of 59 natural watercress beds in the Limousin region (central France) was surveyed over a 15-year period (1990-2004) to detect the contamination of watercress by the metacercariae of Fasciola hepatica and to determine the presence of larval forms in the two species of lymnaeids which live in these waterholes in June and July. The number of beds contaminated with F. hepatica metacercariae varied over the years, and the burden of the larvae on plants was low: a mean of 2.6-6.3 per bed. The same variability was also noted for natural infections of Galba truncatula with F. hepatica, as the annual prevalences ranged from 1.2% to 2.4%. Natural infections of Omphiscola glabra with F. hepatica were only detected from 1996 and the annual prevalences subsequently increased up to 1.4-1.8% between 2001 and 2004. However, for both lymnaeids, the variations in these prevalences with year were insignificant. The contamination of these beds with F. hepatica over the past 15 years was similar to that recorded in the same sites between 1970 and 1986. The main changes were the appearance of another digenea, Paramphistomum daubneyi, in the beds, and the possibility for O. glabra to naturally sustain the larval development of F. hepatica.

Animals↗
PubMed1979

A controlled evaluation of albendazole against natural infections with Fasciola hepatica and Fascioloides magna in cattle.

Albendazole administered to cattle at doses of 45, 35, 25, and 15 mg/kg of body weight reduced the number of viable Fasciola hepatica adults by 99.8%, 97.6%, 97.6%, and 95.7%, F hepatica immatures by 100%, 98.6%, 98.6% and 91.9% and Fascioloides magna by 96.5%, 99.1%, 94.0%, and 94.0%, respectively. Statistical evaluation by one-way analysis of variance, showed significant (P less than 0.05) differences in burdens of each fluke species between treated and untreated cattle, but differences in efficacy between any two of the four drug doses were not significant. Signs of toxicosis were not observed between the time of drug administration and slaughter 7 to 10 days later.

Animals↗
PubMed1992

[Geopolitical distribution of the occurrence of Fasciola hepatica in Santa Catarina State, Brazil].

During 12 years feces samples from cows, water buffaloes, sheeps and goats were examined by sequential tamis filtration to show the occurrence of Fasciola hepatica eggs. The material came from 129 municipalities of Santa Catarina State, and 5 g of feces per animal were examined. The occurrence of F. hepatica was confirmed in 64.82% of the municipalities. Considering the host, F. hepatica was confirmed in goats from Florianópolis, São José, São João Batista and Guaramirim municipalities; in sheep from Brusque, Pomerode, Palhoça and São José; in water buffaloes from 9 and in cows from 86 municipalities. For this study, 13,762 feces samples were examined and in 3,814 the presence of eggs of F. hepatica was demonstrated. The percentage of occurrence for host species was 27.86 in cows, 24.72 in water buffaloes, 16.92 in sheep and 15.66 in goats. By the results it was demonstrated that Itajaí Valley at Southeast Hidrographic Basin, in Santa Catarina State is an endemic area of F. hepatica, even though Uruguai Hidrographic Basin was not referred as a geographical record for this parasite.

Animals↗
PubMed1980

The selective adherence of rat eosinophils to newly excysted Fasciola hepatica in vitro.

Eosinophils present in a mixed population of peritoneal cells, derived from either normal or fluke infected rats, selectively adhered to the tegument of newly excysted Fasciola hepatica in vitro in the presence of immune serum. Adherence occurred independently of complement, was not affected by the age of the sensitising infection and could not be induced by artificially raised antisera to dead fluke antigens.

Animals↗