Comparison of the use of secretory and somatic antigens in an ELISA for the serodiagnosis of hypodermosis.

An antigen was prepared from metabolic products which were produced by maintaining first instar larvae of Hypoderma lineatum in RPMI tissue culture medium for 48 h. Three major proteins were identified in the secretory products and were characterised in terms of their molecular weights and iso-electric points. The antigen compared favourably with a soluble extract of larvae when used in an enzyme-linked immunosorbent assay (ELISA) against a panel of control sera and 2000 bovine sera collected from farms in Andalucia, Spain.

Progress in the British hypodermosis eradication programme: the role of serological surveillance.

The British warble fly eradication programme has resulted in the numbers of infested cattle falling from an estimated four million in 1978 to zero in 1991. In 1982, when about 705 herds were infested the disease was made notifiable, and serological surveillance was begun in 1988. In spring 1991, 227,000 blood samples were tested by using the ELISA technique and no warbled animals were found. In addition no clinical cases were reported during 1991.

Use of an ELISA for serodiagnosis of parasitic bronchitis in cattle.

The enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of parasitic bronchitis in cattle was evaluated in naturally injected, experimentally infected and vaccinated animals. Significant antibody titres could be demonstrated from five weeks after animals had been exposed to the parasite, so that infected animals could be identified during the summer. The test did not give false positive results in vaccinated animals. The technique proved particularly useful in revealing latent infections in milking herds in the autumn when heavy burdens of worms in the lungs do not generate any larvae in the faeces.

Comparative evaluation of enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of dourine.

The detection of antibodies against Trypanosoma equiperdum in 689 equid sera was compared by enzyme-linked immunosorbent assay (ELISA), the complement fixation test (CFT) and an indirect immunofluorescent test (IIF). CFT was the least sensitive technique, susceptible to anti-complementary factors and the most technically demanding. IIF was more sensitive, but was only suitable for testing limited numbers of samples. In this study, ELISA was the most sensitive test, the least labour intensive and lends itself to a considerable degree of automation. It is suggested that ELISA would be relatively easy to standardise between laboratories and an ELISA protocol could be adopted as the internationally approved test for equine health certification.

The serological incidence of Hypoderma bovis in cattle in England and Wales in spring 1989.

Sera from 152,434 cattle held on 6175 farms in England and Wales were tested for the presence of antibodies to Hypoderma bovis during the 10 weeks starting on February 6, 1989. Thirty-nine positive animals (25.6 animals/100,000 tested) were identified on 21 of the farms (0.34 per cent). In comparison, the survey in 1988 examined 74,502 cattle on 3087 farms and found 29 positive animals (38.9/100,000 tested) on 18 farms (0.58 per cent).

Serological survey of the incidence of Hypoderma bovis in cattle in 1988.

Sera from 74,502 cattle from 3087 farms in England and Wales were tested for the presence of antibodies against Hypoderma bovis in the spring of 1988. Twenty-nine positive sera were identified on 18 premises and these animals were treated; an examination of 6030 sera taken from 108 neighbouring herds identified another 17 seropositive animals on 10 farms in Devon, Cornwall, Lancashire, Shropshire and Powys, indicating that these counties still harbour populations of warble fly.

Sero-diagnosis of Fasciola hepatica infections in cattle.

An antigen was prepared from metabolic products which were produced by maintaining Fasciola hepatica adults in RPMI tissue culture medium for 24 h. The antigen compared favourably with a soluble extract of adult flukes when used in an enzyme-linked immunosorbent assay (ELISA). Experimentally infected calves were used to evaluate the sensitivity of the test and a survey from areas of both traditionally high and low incidence of the disease was carried out. Evidence is presented which suggests that this metabolic antigen could be used for the sero diagnosis of naturally occurring infections. In addition the use of a microcomputer to read, file and analyse the results enabled a very large number of samples to be processed daily.

Enzyme-linked immunosorbent assay for the detection of antibodies to the sheep scab mite Psoroptes ovis.

An enzyme-linked immunosorbent assay has been shown to detect antibodies in the circulation of sheep infected with Psoroptes ovis. Strong positive reactions were obtained from 19 sheep with four-month-old infections. No cross reaction was observed with sera obtained from sheep infected with either Fasciola hepatica, Nematodirus battus, Ostertagia circumcincta or Damalinia ovis.

The serum antibody response of infected rats, rabbits, lambs and calves to Fasciola hepatica adult antigen fractions separated by preparative flatbed iso-electrofocusing.

Electrofocusing of F. hepatica adult antigen in granulated gels separated the material into 22 arbitrary fractions. Polyacrylamide gel analysis demonstrated groups of proteins with similar iso-electric points in 19 of the fractions. A microplate ELISA detected antigens in all 22 fractions and was used to test the serum antibody response in infected rats, rabbits, lambs and calves to these antigen fractions. The results indicated that rat and calf sera gave a much stronger response than rabbit and lamb sera to the antigens which separated above pH 7.0. It is possible that the greater efficiency of the rat and bovine immune systems in combating re-infection with F. hepatica may be related to this response.

Demonstration of IgA-containing cells in the small intestine of lambs infected with Nematodirus battus.

A peroxidase anti-peroxidase immunohistochemical technique was used to demonstrate IgA-containing cells in the small intestine of lambs infected with Nematodirus battus. These cells were more numerous in the infected lambs than in the uninfected animals. The difference was greatest for the first three sites, 1 to 3 m distal to the pylorus, where the preponderance of the N battus infection is found. It is suggested that future immunohistochemical studies on the relationships between IgA and resistance to N battus should be directed to this area.

Persistence of antibody in calves after an infection with Hypoderma bovis.

The antibody titre of seven calves, naturally infected with Hypoderma bovis, was estimated by an enzyme-linked immunosorbent assay (ELISA) on sera collected from June to early December 1983 following infection in 1982. It was found that the antibody was lost from the circulation of these animals about 14 weeks after the disappearance of the infection. It is suggested that the presence of antibody in the sera of calves sampled in the late autumn and winter indicated that a new infection had been acquired.

Enzyme-linked immunosorbent assay for the detection of antibodies to Hypoderma bovis in cattle.

An adaptation of the enzyme-linked immunosorbent assay has been shown to detect antibodies in the circulation of cattle infected with Hypoderma bovis. Strong positive reactions were obtained from all infected cattle, even those which harboured only one larva. A strong cross reaction was observed in sera taken from cattle infected with H lineatum, but not in cattle infected with Fasciola hepatica or Ostertagia ostertagi.