[Present Situation of Contagious Bovine Pleuropneumonia in Central African Republic]. / Situation Actuelle de la Péripneumonie Contagieuse Bovine en République Centrafricaine.

Aims: The aim of this study was to analyse the current situation of Contagious Bovine Pleuropneumonia (CBPP) in the Central African Republic (CAR) by seroprevalence analysis, as well as isolation and characterization of strains of the etiologic agent, Mycoplasma mycoides mycoides (Mmm), circulating in livestock breeding regions. Material and methods: The strains obtained were subjected to whole genome sequencing by Illumina technology and genotyped using the eMLST technique based on 62 genes of the Mmm core genome. Their sensitivity to tetracycline was assessed by determination of the minimum inhibitory concentration (MIC) on agar. A seroprevalence analysis by competitive ELISA was conducted in livestock breeding regions (West, Centre and East CAR), including both zebu and taurine cattle breeds, and both males and females. Results: The three strains isolated in the three regions of the CAR shared exactly the same genomic sequence. Phylogenetic analysis showed that they were closely related to a strain isolated in the CAR in 1991, also sequenced in this study, and clustered with Mmm strains originating from East and Central Africa. The recent isolates presented increased MIC values, though they were still sensitive to tetracycline. The global CBPP prevalence in the CAR was estimated at 12.5% with no significant differences observed between cattle breeding regions, nor between males and females. However, a significantly higher prevalence was observed in zebu compared to taurine cattle, most likely in relation to their herding system based on cattle transhumance and nomadic pastoralism. Conclusion: CBPP is enzootic in the CAR in spite of control campaigns based on use of the live T1 vaccines, which have shown little efficacy due to poor implementation in the field. New strategies combining controlled use of antibiotics and inactivated vaccines, with increased thermostability, should be well received by livestock keepers and allow a better control of CBPP in the region. The fact that the recent Mmm isolates are still resistant to tetracycles is encouraging.

Detection of contagious caprine pleuropneumonia in East Turkey.

A study was implemented to investigate the presence of contagious caprine pleuropneumonia (CCPP) in East Turkey. This study was based on clinical surveillance in the field, surveillance at regional slaughterhouses and regular submission of suspected lesions to regional laboratories. The results showed that the agent of CCPP, Mycoplasma capricolum subspecies capripneumoniae (Mccp), could be detected by culture and specific polymerase chain reaction from 37.5% (12/32) of lung samples taken from goats of ten different herds. This agent was also isolated from two of 13 sheep samples (one from the lung and the other from a nasal swab). Mycoplasma capricolum subsp. capripneumoniae was isolated in pure culture and characterised at a finer molecular level. The East Turkish isolate was found to be closely related to another strain of Turkish origin, as well as to Mccp strains isolated in Tunisia. The isolation of Mccp from sheep lung lesions brings the strict host-specificity of this pathogen into question. It may also indicate that Mccp presents a risk for wildlife in the region. Such results, the authors believe, demonstrate that adequate risk assessments should be undertaken in Turkey and neighbouring countries.

Vaccination against contagious bovine pleuropneumonia and the use of molecular tools in epidemiology.

Contagious bovine pleuropneumonia is a serious threat to cattle not only in Africa but also in southern Europe and possibly Asia. It is now present in countries that had been free of the disease for many years, giving rise to doubts about the efficiency of the control strategies. In Africa CBPP is controlled mainly by a vaccination policy that uses variant strains of Mycoplasma mycoides subsp mycoides biotype SC, called T1/44 or T1sr. Until recently, it was not possible to differentiate the various strains within the biotype and consequently to identify the vaccine strains. Restriction analysis of mycoplasma DNA with HindIII and Pst1 has been applied to 24 strains of African origin and one European strain. Each enzyme gave rise to different restriction profiles and the combination of the results permitted subdivision of these strains into 9 groups. Interestingly, some profiles of pathogenic strains seem to be restricted to certain geographical areas. The profile of the poorly immunogenic vaccinal strain KH3J is also very peculiar, and it is easily distinguished from that of the other vaccine strains originating from T1. This technique is simple once the strains are isolated. Efforts are now under way to use molecular tools based on PCR products to alleviate the difficulty of isolation.

Comparison between c-ELISA and CFT in detecting antibodies to Mycoplasma mycoides mycoides biotype SC in cattle affected by CBPP in Botswana.

Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides mycoides biotype small colony (SC) (MmmSC) appears to be making a serious comeback in Africa after successful control programs in many parts of the continent during the 1960s and 1970s. Botswana, a country that has been free from the disease for more than 50 years, was affected in 1995. An eradication policy was adopted by the Government of Botswana in which 320,000 cattle in the affected district of Ngamiland, Northwestern Botswana were slaughtered. This was followed by a restocking exercise in which 70,000 cattle were sent to the outbreak areas as replacement stock. It became necessary to carry out serosurveillance in order to ensure that the disease did not reenter Botswana and to ensure that the replacement stock remained free from the disease. The specificity and sensitivity of the complement fixation test (CFT) in Botswana was assessed in 82 cattle affected by the disease and held in a double fenced quarantine camp. The newly developed competitive ELISA was made available to the National Veterinary Laboratory (NVL) through the FAO/IAEA Joint Division in Vienna, Austria. Using postmortem lesions as the gold standard and a 2 x 2 contingency table, the two tests were compared in terms of their sensitivity and specificity in detecting antibodies to MmmSC. The CFT was found to be slightly more sensitive than the c-ELISA, and this could be related to the stage of the disease. A long-term study comparing the progression of the disease with the two tests is, therefore, essential.

Contagious bovine pleuropneumonia. A reassessment of the efficacy of vaccines used in Africa.

Contagious bovine pleuropneumonia is a major threat for cattle in Africa. Since 1956 the T1/44 strain has been used as a vaccine, and later on, T1sr, a streptomycin-resistant variant that gives fewer post-vaccinal reactions had been developed. These vaccines are known not to be very efficient but they normally should provide protection for about eight months. However, recent emergency vaccinations, performed in various countries in the southern part of the continent apparently met with failure, casting doubts on the identity as well as the protection afforded by the T1sr strain. A vaccine trial has been designed to reassess the real protection afforded by these vaccines in face of recently isolated pathogenic strains. Great care has been taken to test the original vaccinal strains at a dose corresponding to the minimum requirement by international standards. The test was performed in Cameroon, Kenya, and Namibia as to take into account the genetic diversity that exists among the pathogenic strains. In those conditions, the protection rate at three months varied from 33 to 67%, whatever the strain used, T1/44 or T1sr. These results call for additional research for vaccine development and careful planning of strategies in the fight against CBPP.

A competitive ELISA for the specific diagnosis of contagious bovine pleuropneumonia (CBPP).

A competitive ELISA, using a specific monoclonal antibody, was designed to detect antibodies to Mycoplasma mycoides subsp. mycoides SC, the agent of contagious bovine pleuropneumonia. One monoclonal antibody was found suitable for such a test, '117/5', it does not cross-react with any of the other mycoplasma species tested, furthermore, its binding is inhibited by positive sera. The cutoff, 50% of inhibition, was determined using a set of negative sera from CBPP-free areas. The sensitivity was controlled with sera from artificially infected animals as well as from sera from areas where CBPP is enzootic. In both cases, cELISA compared favorably with CFT. The precocity of detection was similar but cELISA detected more positives and the positive titers seemed to persist longer than in the case of CFT. Lysis of the antigen used to coat the ELISA plates reduced the variability of fixation and improved the repeatability of the test. A field evaluation is now in progress which will determine the true sensitivity and specificity of the test and also check if antibodies are detected after vaccination.

Phylogeny of the Mycoplasma mycoides cluster as shown by sequencing of a putative membrane protein gene.

The Mycoplasma mycoides cluster is made of six species that are closely related both genetically and phenotypically. Two are of particular importance, M. mycoides subsp. mycoides SC causing contagious bovine pleuropneumonia and M. capricolum subsp. capripneumoniae causing contagious caprine pleuropneumonia. The sequences of a putative membrane protein gene and partial flanking open reading frames have been obtained from various strains in this cluster, including all reference strains. Sequence analysis showed this locus is present and fully conserved in all strains of M. mycoides subsp. mycoides SC isolated from geographically most distant places worldwide. In M. capricolum subsp. capripneumoniae polymorphism in this locus has been found at seven positions and revealed that they can be used as epidemiological markers. Conserved regions were used to define a primer pair that enables the amplification by PCR of two fragments 302 and 1298bp long, respectively. The 302bp long fragment contains an intergenic sequence that can be used for phylogenetic studies or for identification purposes. Parsimony analysis on an alignment of 49 DNA sequences show a subdivision of the M. mycoides cluster into two subgroups that is in accordance with results obtained by phenotypic methods. Two lineages exist within the capricolum subgroup, one of them clustering strains identified as M. capricolum subsp. capricolum, M. capricolum subsp. capricolum and M. sp Bovine Group 7. However M. capricolum subsp. capripneumoniae strains can readily be identified by three specific nucleotide positions or by sequencing the 1298bp long fragment. There is no clear subdivision within the mycoides subgroup, supporting the idea that M. mycoides subsp. mycoides LC and M. mycoides subsp. capri should not be separated into two subspecies. Mycoplasma mycoides subsp. mycoides SC strains can easily be distinguished as they bear an insertion sequence 15bp downstream from the stop codon of the membrane protein gene.

A specific PCR for the identification of Mycoplasma capricolum subsp. capripneumoniae, the causative agent of contagious caprine pleuropneumonia (CCPP).

Contagious caprine pleuropneumonia is a severe infectious disease of goats in Africa and the Middle East. It is caused by a fastidious mycoplasma, Mycoplasma capricolum subsp. capripneumoniae, a member of the "M. mycoides cluster". Members of this cluster share genomic and antigenic features, which result in common biochemical and serological properties, complicating species identification. Two species of this cluster, M. mycoides subsp. capri and M. mycoides subsp. mycoides large colony biotype, are very often isolated from clinical cases resembling contagious caprine pleuropneumonia. Furthermore, in the laboratory, M. capricolum subsp. capripneumoniae can be easily confused with the closely related capricolum subspecies. Considering these constraints and the scarcity of available methods for identification, a specific polymerase chain reaction was developed. A DNA fragment of 7109 bp containing genes coding for the arginine deiminase pathway (ADI) was chosen as target sequence for the selection of a specific primer pair. The full ADI operon from M. capricolum subsp. capripneumoniae strain GL100 was sequenced. Polymorphism within this locus was analyzed by comparison with the sequence from the closely related IPX strain (M. capricolum subsp. capricolum). It varied from 0.6% to 3.5%. The highest divergence was found in a region coding for arcD. Therefore, this gene was chosen as target for the specific amplification of a 316 bp-long DNA fragment. The specificity of this PCR was validated on 14 M. capricolum subsp. capripneumoniae strains and 27 heterologous strains belonging to the "M. mycoides cluster" and M. putrefaciens. This new PCR will be a valuable tool for the surveillance of contagious caprine pleuropneumonia.

Molecular epidemiology of contagious bovine pleuropneumonia by multilocus sequence analysis of Mycoplasma mycoides subspecies mycoides biotype SC strains.

Contagious bovine pleuropneumonia is a bacterial disease caused by Mycoplasma mycoides subsp. mycoides SC (MmmSC), and included in list A of the Office International des Epizooties. It is one of the major constraints to cattle raising in sub-Saharan and south-western Africa and also a threat to all countries currently free of the disease. MmmSC strains were considered very homogeneous until 1995, when various techniques such as enzymatic restriction of whole DNA or Southern blotting showed that this was not the case. These techniques are unfortunately difficult to standardize and require the extraction of DNA from an MmmSC culture. We therefore decided to investigate the possibility of constructing a molecular epidemiology tool based on multilocus sequence analysis (MLSA) with PCR amplification of various loci followed by sequencing. Six loci were found suitable for this purpose and an additional PCR was designed to detect the presence of an 8.8kb deletion described by others in some strains. Fifteen different MLSA profiles were evidenced in our study. They allowed a clear distinction between European, south-western African and sub-Saharan strains. In addition, the results obtained on strain PO1967 confirmed its European origin, even though it does not exhibit the 8.8kb deletion. This new tool for contagious bovine pleuropneumonia may prove particularly useful for identifying MmmSC strains in countries at risk from contamination. It can also easily be refined by adding more strains or other loci of interest.

Comparative analysis of the lppA locus in Mycoplasma capricolum subsp. capricolum and Mycoplasma capricolum subsp. capripneumoniae.

The lppA gene, encoding the lipoprotein named LppA[Mcaca] was characterised in Mycoplasma capricolum subsp. capricolum. It encodes a lipoprotein with an apparent molecular mass of 57 kDa as determined by SDS-PAGE. Using antibodies directed against recombinant LppA[Mcaca], we showed the expression of this lipoprotein in all M. capricolum subsp. capricolum by immunoblot analysis. The serum did not cross-react with other members of the Mycoplasma mycoides cluster, hence showing that LppA[Mcaca] was antigenically specific to M. capricolum subsp. capricolum. The lppA gene was conserved within the subspecies and was used for the development of a specific PCR assay for the identification of M. capricolum subsp. capricolum. The taxonomically related Mycoplasma capricolum subsp. capripneumoniae (F38) was found to contain an lppA-pseudo-gene. It showed high similarity to functional lppA genes of other mycoplasmas in the M. mycoides cluster. However, it contained interrupted open reading frames. Moreover, the nucleotide sequence of the lppA pseudo-genes in different strains of M. capricolum subsp. capripneumoniae were quite variable. Interestingly, the lppA pseudo-gene had a size similar to that of the functional lppA genes of other mycoplasmas of the M. mycoides cluster and occupied the same genomic location as the latter ones in the vicinity of the mtlD genes. This study showed that all members of the M. mycoides cluster contain each a species-, subspecies- respectively type- specific lppA gene analogue which encodes a lipoprotein that has structural and functional relationship to the surface lipoprotein LppA [MmymySC], previously named P72, of M. mycoides subsp mycoides SC, with the exception of M. capricolum subsp. capripneumoniae which seems not to express an LppA analogue.

Phage displayed peptides and anti-idiotype antibodies recognised by a monoclonal antibody directed against a diagnostic antigen of Mycoplasma capricolum subsp. capripneumoniae.

A monoclonal antibody (Mab 4.52) raised against Mycoplasma capricolum subsp. capripneumoniae (Mccp) cell lysate was used as a template to obtain substitute antigens recognised by its paratope. Two approaches were investigated: a 17-mer random peptide library displayed on the surface of a filamentous phage was screened by panning on the immobilised Mab 4.52 and anti-idiotype antibodies were generated by immunising a chicken with the F(ab')(2) fragments of the antibody. Analysis of the peptide sequences displayed by the isolated phages identified two peptides. Both contained two cysteine residues and had identical or similar amino acids in positions 5 (P), 8 (I/L) and 13 (L). The fusion phages were also recognised by Mab 4.52 in enzyme-linked immunosorbent assay (ELISA) and binding was shown by surface plasmon resonance. One of the peptides was a markedly better inhibitor (67%) of the binding of Mab 4.52 to its original antigen than the other (20%) at 1mg/ml. After absorption, to remove isotypic and allotypic reactivities, the anti-idiotype IgY was specifically recognised by Mab 4.52 in ELISA and was able to inhibit its binding to the original antigen, whereas anti-idiotype antibodies raised against a bluetongue virus-specific antibody had no effect. In spite of unequivocal binding of the anti-idiotype antibodies and the fusion phages to the paratope of Mab 4.52, goat antisera appeared not to react with either of the surrogate antigens. In contrast, the test sera bound to the original antigen suggesting that Mab 4.52 does not recognise exactly the same antigenic site as antibodies in the goat antisera.

The use of monoclonal antibodies in the diagnosis of contagious caprine pleuropneumonia (CCPP).

Contagious caprine pleuropneumonia is a severe disease affecting goats in Eastern Africa and the Middle East, caused by Mycoplasma sp. type F38. Its exact geographical distribution is however not exactly known due to the lack of specificity of the available serological tests and the difficulty in cultivating M. sp. F38. A panel of monoclonal antibodies (mAbs) was produced, using crude or membrane proteins antigens from type F38 strains to immunize mice. The reactivity of the mAbs was tested by an immunobinding assay with crude mycoplasma antigens spotted on nitrocellulose filters. One hundred and twelve antigens, standardized at 0.5 mg protein/ml, were used. Mycoplasma strains were chosen among closely related species of the "mycoides cluster", M. capricolum, Group 7 of Leach, M. mycoides mycoides LC, M. mycoides mycoides SC, M. mycoides capri, as well as among species that are isolated from goat lungs, M. arginini, M. ovipneumoniae, M. putrefaciens, M. agalactiae. Out of 60 mAbs, 4 were chosen to build an identification test for mycoplasmas of the "mycoides cluster". Controls showed that accurate identification could be hampered by antigenic heterogeneity within the M. capricolum species. One mAb was used for the direct detection of M. sp. F38 antigen in pleural fluid from goats suspected of CCPP. The sensitivity of the test can be estimated at 0.5 micrograms protein/ml. Comparison with isolation results show a 74% agreement between the two methods. The same mAb was used to build a blocking ELISA. This serological test was strictly specific for CCPP. It detects antibodies in sera of naturally infected or artificially immunized animals while it remained negative with hyperimmune sera to related strains such as PG 50. Direct antigen detection and blocking ELISA are tools that may enable a better assessment of CCPP distribution.

Genetic evolution of Mycoplasma capricolum subsp. capripneumoniae strains and molecular epidemiology of contagious caprine pleuropneumonia by sequencing of locus H2.

Contagious caprine pleuropneumonia (CCPP) is a major threat to goat farming in developing countries. Its exact distribution is not well known, despite the fact that new diagnostic tools such as PCR and competitive ELISA are now available. The authors developed a study of the molecular epidemiology of the disease, based on the amplification of a 2400 bp long fragment containing two duplicated gene coding for a putative membrane protein. The sequence of this fragment, obtained on 19 Mycoplasma capricolum subsp. capripneumoniae (Mccp) strains from various geographical locations, gave 11 polymorphic positions. The three mutations found on gene H2prim were silent and did not appear to induce any amino acid modifications in the putative translated protein. The second gene may be a pseudogene not translated in vivo, as it bore a deletion of the ATG codon found in the other members of the "Mycoplasma mycoides cluster" and as the six mutations evidenced in the Mccp strains would induce modifications in the translated amino acids. In addition, an Mccp strain isolated in the United Arab Emirates showed a deletion of the whole pseudogene, a further indication that this gene is not compulsory for mycoplasma growth. Four lineages were defined, based on the nucleotide sequence. These correlated relatively well with the geographical origin of the strains: North, Central or East Africa. The strain of Turkish origin had a sequence similar to that found in North African strains, while strains isolated in Oman had sequences similar to those of North or East African strains. The latter is possibly due to the regular import of goats of various origins. Similar molecular epidemiology tools have been developed by sequencing the two operons of the 16S rRNA gene or by AFLP. All these various techniques give complementary results. One (16S rRNA) offers the likelihood of a finer identification of strains circulating in a region, another (H2) of determining the geographical origin of the strains. These tools can make a very useful contribution to understanding the epidemiology of CCPP.

ISCOM vaccine against contagious bovine pleuropneumonia (CBPP). 1. Biochemical and immunological characterization.

A better vaccine than the existing ones against contagious bovine pleuropneumonia (CBPP) caused by Mycoplasma mycoides subsp. mycoides small colony type (MmmSC) would improve the chances for eradication of CBPP. In such an effort, immunostimulating complexes (ISCOMS) have been prepared from the whole detergent-solubilized cells of MmmSC and characterized biochemically and immunologically. The most efficient detergent for solubilization of the mycoplasma was MEGA-10 which yielded a high recovery of proteins in the ISCOMS. The ISCOMS showed the typical cage-like structure by EM and sedimented as 19S by sucrose gradient centrifugation. The protein pattern of the ISCOMS, analyzed in SDS-PAGE, revealed a great number of bands distributed along the gel as high and low molecular weight polypeptides. The Western blot developed with a serum from a CBPP infected animal detected a reduced number of polypeptides. In samples from whole mycoplasma cells and in ISCOMS, lectin blots revealed more than 20 carbohydrate structures. The ISCOMS induced a strong primary antibody response in mice measured by ELISA and the boost resulted in a 6-fold increase of the serum antibody response. The IgG response was distributed into various IgG subclasses with high IgG1, IgG2a and IgG2b titres while the IgG3 response was low. In cattle the ISCOM vaccine induced strong primary and long lasting secondary antibody responses of similar magnitudes as those of naturally infected animals as recorded by ELISA which persisted more than a year. IgG response was equally distributed in IgG1 and IgG2 subclasses. Also a cell-mediated immune response measured by proliferation assay was induced by low dose of ISCOMS. In the growth inhibition test, sera from vaccinated cattle readily inhibited colony growth already after the first immunization.

Contagious bovine pleuropneumonia vaccines and control strategies: recent data.

Contagious bovine pleuropneumonia is one of the most threatening transboundary cattle disease in Africa. However, with the exception of Botswana, very few African countries were able to implement eradication strategies for this disease, after it had recently re-infected a number of countries. Previous experimental studies have shown that emergency vaccination campaigns, based on a single injection, were not inducing a sufficient protection level to prevent further spread of the disease. In addition, post-vaccinal reactions were sometimes reported in the field when using vaccine strain T1/44, leading cattle owners to refuse the vaccination. On the contrary, antibiotics are used quite often in the field but there are insufficient data to assess their efficacy properly. Therefore experimental studies were implemented: (i) to check if higher dosages of the vaccine would be able to induce higher protection rates and (ii) to elucidate the origin of the post-vaccinal reactions observed with T1/44 and (iii) to gain preliminary results on the efficacy of long-acting tetracycline. The first experiment included the use of three doses of vaccine strains T1/44 and T1sr: 10(7), 10(8) and 10(9) mycoplasmas per dose. T1/44 seemed to induce a higher protection (70%) than T1sr (60%). However, there was no observable dose effect for these vaccine strains. The second experiment was performed by injecting various MmmSC strains subcutaneously into susceptible cattle. One of these strains was an isolate obtained from a "Willems" reaction following a vaccination with T1/44. This isolate, called T1B, induced typical invading oedema at the injection site in a similar way to the pathogenic strain, whereas the original T1/44 vaccine strain did not. These findings indicate that the strain has reverted to virulence. Finally the antibiotic trials showed that long-acting tetracycline was able to reduce the losses due to the disease but could not prevent the persistence of viable MmmSC in treated animals. The consequences of these findings are discussed. They reinforce the need for additional research on new vaccines able to elicit longer lasting protection. However, once continuing additional field research is obtained, it should allow better defined strategies to be put in place. Meanwhile, immediate action should be taken to prevent the further spread of CBPP in the Southern part of Africa.

Contagious bovine pleuropneumonia vaccines, historic highlights, present situation and hopes.

Contagious bovine pleuropneumonia (CBPP) is a contagious infection of cattle caused by a mycoplasma, M. mycoides subsp. mycoides SC (MmmSC). It induces lesions of pleuropneumonia in acute cases and the formation of pulmonary "sequestra" in chronic cases. The disease is prevalent mostly in Africa, where it is responsible for high losses, but it has also been sporadically present in Southern Europe until 1999. Vaccination is now prohibited in most countries except in Africa. An empirical "inoculation" procedure was developed as early as 1852 in Europe but it may have been used even earlier in Africa. The inoculation of pleural fluid was performed at the tip of the tail in Europe and on the bridge of the nose in Africa. It conferred good protection but induced a high number of fatal cases. Various inactivated preparations have been tested in the past with inconclusive results leading sometime to some protection and some other time to a sensitisation of the immunised animals. Such preparations have never been used in the field. Attenuated MmmSC strains have been developed in the 1950s and used extensively in the field both in Africa and Australia. The best known vaccine strains are KH3J, T1/44 and T1sr. Vaccination campaigns have succeeded in reducing considerably the CBPP prevalence in these two continents but eradication was achieved in Australia only by switching to strict measures of animal movement control and a stamping-out policy. The search for new CBPP vaccines has become a major issue for African countries that are facing an increase in outbreaks. The rationale for this search is based on a better understanding of the mycoplasma virulence mechanisms that could lead to a targeted attenuation of MmmSC strains. It is also based on a better understanding of the bovine immune response that may be driven to a pathogenic inflammatory response or conversely to a better balanced response leading to protection.

Contagious caprine pleuropneumonia and other pulmonary mycoplasmoses of sheep and goats.

Contagious caprine pleuropneumonia (CCPP) is now a well-defined disease that is caused by Mycoplasma capricolum subsp. capripneumoniae. CCPP is infectious, contagious and fulfils the classic Koch postulates that characterise such types of disease. The distribution of the disease is not exactly known, but reports of mycoplasma isolation and official declarations to the Office International des Epizooties (OIE) enable a probable distribution map to be obtained. There are many other mycoplasmas that can infect goat and sheep lungs and induce pleuropneumonia. However, pleuropneumonia is often restricted to young animals and the prominent symptom is mastitis in lactating does. Other symptoms may also occur, contributing to a syndrome that has been tentatively described in this paper as 'MAKePS syndrome' for mastitis, arthritis, keratitis, pneumonia and septicaemia.

[Diagnosis of caprine contagious pleuropneumonia: recent improvements]. / Diagnostic de la pleuropneumonie contagieuse caprine: améliorations récentes.

Due to the difficulty in isolating Mycoplasma sp. type F38, two techniques for rapid detection are proposed in order to identify the presence of mycoplasmas of the mycoides group in samples of pleural fluid. A modification in the composition of isolation media promotes the growth of type F38 mycoplasmas and inhibits the growth of M. ovipneumoniae strains.

Diagnosis and control of contagious caprine pleuropneumonia.

The diagnosis of contagious caprine pleuropneumonia (CCPP) has often been considered difficult. This is because of the confusion that can arise with other mycoplasmoses of small ruminants. Symptoms and lesions can be similar and the isolation of M. capricolum subsp. capripneumoniae (MccF38) requires skilled technicians. Once MccF38 strains are isolated, their identification should not be difficult. New techniques, such as polymerase chain reaction, now offer the possibility of identifying MccF38 directly from dried samples. However, the isolation of MccF38 strains is always required for an official declaration of infection. Until now, the official serological test has been the complement fixation test; the main drawbacks being lack of sensitivity and specificity and also the short persistence of antibodies detected by this technique. The specific competition enzyme-linked immunosorbent assay has now been developed and should enable wide serological enquiries to determine the real prevalence of the disease. Antibiotic treatments are effective but may not prevent persistence in latent carriers. An inactivated vaccine with saponin as an adjuvant has been produced in Kenya, which protects goats for approximately one year.

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.