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Biomedical subjects

L Dedieu

Publications and source records attributed to L Dedieu.

5 recordsLinked to original sources

Contagious bovine pleuropneumonia vaccines: the current situation and the need for improvement.

The control of contagious bovine pleuropneumonia (CBPP) has been clearly identified by the Organisation of African Unity/Inter-African Bureau of Animal Resources as a priority. In the first part of this article, the authors introduce the past and present vaccines, based on the two classic strains, T1, and KH3J. They describe the guidelines for vaccine production technology, and the quality control requirements for CBPP vaccines of the Office International des Epizooties. The failure of the currently used T1-SR vaccine to provoke satisfactory immunity in cattle, particularly in the newly infected areas of Africa, is pointed out. Other shortcomings of the current CBPP vaccines are also highlighted. Thus, there is a need to improve CBPP vaccines and the authors propose detailed emergency measures to address this problem. In the second part of the article, a subunit approach using immunostimulating complex technology is outlined. The authors emphasise the importance of current research in cell-mediated immunity and immunopathology, which is aimed at improving the efficacy of CBPP vaccines.

Animals

Development of two PCR assays for the identification of mycoplasmas causing contagious agalactia.

A new detection test for the mycoplasmas causing contagious agalactia, Mycoplasma agalactiae, M. capricolum subsp. capricolum and M. mycoides subsp. mycoides L. C., was developed. It was based on two polymerase chain reaction assays: the Ma-PCR for the detection of M. agalactiae and the MYC-PCR for the 'mycoides cluster' thus including M. capricolum subsp. capricolum and M. mycoides subsp. mycoides L. C. An M. agalactiae strain was identified by a 933-bp Ma-PCR product and no amplification with the MYC-PCR. In contrast, a 460-bp MYC-PCR product and a negative or a 350-bp Ma-PCR product characterized a 'mycoides cluster' strain. M. capricolum subsp. capricolum and M. mycoides subsp. mycoides L. C. were identified by their species-specific AseI pattern of the 460-bp MYC-PCR product.

Mycoplasma

Development of a selective polymerase chain reaction assay for the detection of Mycoplasma mycoides subsp. Mycoides S.C. (contagious bovine pleuropneumonia agent).

A new selective assay for the detection of Mycoplasma mycoides subsp. mycoides SC (MmmSC) via the polymerase chain reaction (PCR) has been developed. This test used two PCR assays: a control-PCR (MYC-PCR) identifying the pathogen as a member of the mycoides cluster and the MSC-PCR which is specific for MmmSC. The MYC primers targeted a DNA sequence of about 460 bp from all the 59 mycoides cluster-strains tested. No amplification occurred with bovine genomic DNA or with the 11 other bacterial species assayed. The MSC primers selectively amplified a 275 bp sequence from the 27 MmmSC strains tested, with three specific internal restriction sites allowing confirmation of the identification. The sensitivity assessed by direct agarose gel analysis for both PCR assays was 100 CFU. The sensitivity of the MSC-PCR was increased to 1 CFU by a dot-blot hybridization step using, as a probe, the entire 275 bp sequence digoxigenin-labeled by PCR. These two PCR assays were successfully used to detect MmmSC in pleural fluids from naturally-infected cattle. We conclude that these two PCR assays may be valuable tools for the diagnosis of contagious bovine pleuropneumonia.

Animals

Field diagnostic kits: a solution for developing countries?

An exact assessment of the animal health situation in a country is an essential element in formulating eradication and control programmes, and in regulating international trade in animals and animal products from that country. Due to a lack of human and technical resources, Veterinary Services in developing countries often lack precise knowledge on disease occurrence. Since the collection and transmission of reliable information on animal diseases in developing countries are major concerns of the Office International des Epizooties (OIE), a project aimed at improving this situation was implemented with international financial support. This project involved the development by the Centre for the Application of Methodology for the Diagnosis of Animal Diseases (CAMDA) of field kits for the diagnosis of the main diseases present in tropical Africa: rinderpest, peste des petits ruminants (PPR), contagious bovine pleuropneumonia (CBPP) and contagious caprine pleuropneumonia (CCPP). Several tests already exist, such as complement deoxyribonucleic acid (cDNA)-specific probes and polymerase chain reaction (PCR) for rinderpest and PPR, DNA probes and PCR for CBPP, capture enzyme-linked immunosorbent assay, the agglutination test and the immunobinding peroxidase test for CCPP, etc. With specific reference to these examples, the various problems faced by the OIE and CAMDA are reviewed.

Animals

Development of a species-specific DNA probe for Mycoplasma capricolum.

A specific DNA probe for the detection and identification of Mycoplasma capricolum, one of the causative agents of contagious agalactia syndrome, was selected from a genomic library. It consists of a 900bp RsaI genomic fragment of M. capricolum (reference strain), cloned into the EcoRV site of the plasmid Bluescript. By using the appropriate stringency this radiolabelled probe reacts specifically with M. capricolum when tested by dot blot hybridization against various mycoplasmal DNAs. The current level of sensitivity of the 32P-labelled 900bp RsaI probe is 500 pg of homologous DNA, corresponding to 5 x 10(4) mycoplasmas. A non radioactive labelling method, using the digoxigenin-11-dUTP, was also tested. The specificity of the digoxigenin-labelled probe was equivalent to that obtained with the radioactive probe. However the sensitivity of detection decreased to 1 ng of homologous DNA detected, corresponding to 1 x 10(5) mycoplasmas. Tests performed with milk samples have demonstrated that the radioactive 900 bp RsaI probe indeed detected M. capricolum contained in milk. A positive signal was obtained when 10(5) M. capricolum were present in the spot.

Animals