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Biomedical subjects

A G Luckins

Publications and source records attributed to A G Luckins.

At least 73 records · Page 4Linked to original sources

Use of antigens of cultured Trypanosoma brucei in tests for bovine trypanosomiasis.

MicroELISA tests for the detection of trypanosomal antibodies in cattle infected with Trypanosoma brucei, T. vivax and T. congolense were carried out using antigens prepared from culture forms and bloodstream forms of T. brucei. Antigens prepared from culture forms gave similar microELISA values to those obtained with bloodstream form antigens and statistical analysis of the results obtained with both antigen preparations and sera from the infected animals showed significant high positive correlation. It would thus be possible to use culture from antigens as an alternative to bloodstream form antigens in diagnostic tests for trypanosomiasis, with the advantage that culture form antigens are more easily prepared free from extraneous protein.

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Evaluation of an enzyme immunoassay for serodiagnosis of infections with Theileria parva and T annulata.

An enzyme linked immunosorbent assay (ELISA) was used to determine antibody levels in cattle infected with Theileria parva and T annulata, using antigens prepared from the intra-erythrocytic piroplasm stage of the parasites. Antibody levels in calves infected with T parva increased from the 16th day after infection to reach peak values at days 28 to 35 and then declined rapidly, but in calves infected with T annulata antibody levels rose steadily up to day 40. Similar patterns of antibody production were shown by indirect fluorescent antibody tests. Sera from animals infected with T parva gave higher ELISA values with the antigen prepared from the homologous parasite species than with the antigen prepared from T annulata, but sera from cattle infected with T annulata gave similar high ELISA values with antigens prepared from both T parva and T annulata. Sera from animals infected with T mutans, T sergenti, T velifera, Babesia divergens, B major and B bovis gave only slight or no cross reactions with the piroplasm antigens, but serum from a calf infected with B bigemina cross reacted at a significant level with both piroplasm antigens.

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Serodiagnosis of infection with Trypanosoma evansi in camels in the Sudan.

Five diagnostic tests for infection with Trypanosoma evansi have been compared in groups of camels experimentally infected or exposed to natural infection in the Sudan. The correlation of positive results obtained by assays of IgM levels, the mercuric chloride test and the formol gel test with the presence of active infection was unsatisfactory, but there was a good correlation between results obtained using IFAT and ELISA and proven infection. Sera from a high proportion of apparently uninfected camels from endemic areas gave positive reactions with all 5 tests, possibly indicating inadequate parasitological diagnosis or persistence of antibody after unsatisfactory chemotherapy. It was concluded that serological tests using trypanosomal antigens to detect antibodies were more sensitive for diagnosis than indirect tests based on raised euglobulin levels. Serodiagnostic tests may therefore have a place in future programmes for surveillance and control of T. evansi infections in camels.

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Observations on the antigenicity and serological relationships of stocks of Trypanosoma congolense from East and West Africa.

Antigenic relationships of 4 stocks of Trypanosoma congolense from different parts of Africa were examined by immunofluorescence (IFAT) and neutralization tests. Antisera to each stock were obtained from rabbits infected with trypanosomes transmitted by Glossina morsitans. Trypanosomes for use as antigens were obtained from local skin reactions developing on rabbits infected with 2 of the stocks. Using the IFAT and antisera at end-point dilutions approximately 40% of the trypanosomes fluoresced strongly and a further 30% less intensely with homologous antisera, indicating antigenic heterogeneity among the trypanosomes developing in the skin. Using antisera at low dilutions some samples gave cross-reactions with trypanosomes of heterologous stocks, but at higher dilutions there were no cross-reactions with either of the antigens. The lack of cross-reactions at high antiserum dilutions was interpreted as indicating antigenic differences between the 4 trypanosome stocks. Using neutralization tests only homologous antisera reduced the infectivity of trypanosome suspensions. Overall, these observations indicated that there were at least 3 different strains of T. congolense among the 4 stocks examined.

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Trypanosomes in the lymph nodes of cattle and sheep infected with Trypanosoma congolense.

The prefemoral lymph nodes of two calves and a sheep infected with a stock of Trypanosoma congolense transmitted by Glossina morsitans were examined histologically for the presence of trypanosomes. Ten days after infection trypanosomes were found in the subcapsular sinuses of the nodes of a calf and the sheep but parasites were absent from the blood at this time. Trypanosomes were also detected in the prefemeral lymph node of the other calf on examination 30 days after infection, when parasites were also present in the blood. These observations provide further evidence that extravascular foci of trypanosomes develop in infections with T congolense and indicate that it should not be regarded as a strict plasma parasite.

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Evaluation of an indirect fluorescent antibody test, enzyme-linked immunosorbent assay and quantification of immunoglobulins in the diagnosis of bovine trypanosomiasis.

An indirect fluorescent antibody test (IFAT), a microscale version of the enzyme-linked immunosorbent assay (microELISA) and determination of IgM levels in serum were assessed for their comparative diagnostic value in the detection of bovine trypanosomiasis. Serum samples from drug-treated N'dama cattle and untreated N'dama and Zebu cattle from Liberia were examined for the presence fo antibodies to trypanosomes. In the untreated Zebu cattle, infections with T. vivax predominated and the prevalence of infection was higher than that found in untreated N'damas in which infections with T. congolense predominated. The proportion of animals which showed serological evidence of trypanosomiasis in the untreated Zebus was slightly higher than that found in the untreated N'damas. The prevalence of infection was low in N'dama cattle which had been treated with diminazene aceturate and homidium chloride but 50% of the animals showed serological evidence of trypanosomiasis. More serologically positive animals were detected by microELISA than IFAT, but both tests were equally sensitive in detecting antibodies in cattle in which trypanosomes were demonstrated by examination of peripheral blood. With both IFAT and microELISA it was necessary to carry out tests using antigens prepared from T. brucei, T. vivax and T. congolense in order to detect all serologically positive animals. Increases in serum IgM occurred in both N'dama and Zebu cattle but the levels were raised in only approximately half of the known infected animals. Overall, more animals gave positive reactions with IFAT and microELISA than showed raised IgM levels.

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Detection of antibodies in trypanosome-infected cattle by means of a microplate enzyme-linked immunosorbent assay.

A micromodification of the enzyme-linked immunosorbent assay (ELISA) was evaluated for its potential application in the immunodiagonsis of bovine trypanosomiasis. Serum samples from infected and non-infected Zebu cattle and samples from Friesian cattle with experimental infections were exmained for the presence of trypanosomal antibodies. There were significant differences between the microELISA values obtained with samples from infected and non-infected cattle. During the course of infection microELISA values were found to fluctuate and the antibody response varied in individual animals. The test did not distinguish between infections with Trypanosoma brucei, T. vivax and T. congolense. There were no cross-reactions between trypanosome antigens and serum samples from cattle infected with T. theileri, Theileria parva, Th. mutans, Th. annulata, Babesia divergens and Anaplasma marginale.

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Observations on a herd of beef cattle maintained in a tsetse area. II. Assessment of the development of immunity in association with trypanocidal drug treatment.

Three groups of Boran cattle were maintained under different trypanocidal drug regimes for 29 months in an area of "medium" tsetse challenge. The development of immunity to trypanosmiasis in the cattle was assessed on the basis of changes in trypanocidal drug requirement, development of parasitaemia, ability to maintain normal blood values, growth and response to challenge after the withdrawal of the drugs. Observations were also made on serum antibody content by immunofluorescence and on serum immunoglobulin levels. Cattle treated with Berenil on the development of clinical disease developed a partial immunity to trypanosomiasis after two years. Cattle treated as a group with Berenil on the development of patent parasitaemia in any one animal of the group did not develop immunity. Cattle treated as a group with Samorin using the same criterion for treatment developed a degree of immunity to the disease. It was concluded that group treatment with Samorin was the most suitable of the three regimes examined for the maintenance of beef cattle is a tsetse infested area.

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The immune response of zebu cattle infection with Trypanosoma congolense and T. vivax.

Zebu cattle were infected with Trypanosoma congolense and T. vivax and changes in the levels of serum immunoglobulins were measured during infection and after treatment. Increases in the levels of IgM occurred within 14 days of infection coincidentally with the first appearance of trypanosomes in the peripheral blood. IgM levels rose to 2-10 times preinfection levels and remained high until the animals were treated with diminazene aceturate. Levels then declined to approximately pre-infection levels within 25 days. IgG levels showed little change. In one animal infected with T. congolense which died of trypanosomiasis, there was an initial increase in IgM levels but subsequently both IgM and IgG levels declined until the animal died. The neutralizing activity of whole serum, and serum IgM and IgG fractions from an ox infected with T. congolense were examined. High levels of neutralizing antibodies against trypanosomes isolated on the eighth day after infection were found in sera collected 12-29 days after infection. Most of the antibody activity was located in IgG fractions which showed a peak in neutralizing activity between day 17 and 22. IgG fractions of the sera showed only slight antibody activity.

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