COST European Cooperation on Scientific and Technical Research European Commission COST 826 Agriculture and biotechnology Mycoplasmas of ruminants: pathogenicity, diagnostics, epidemiology and molecular genetics Volume 3 Edited by L. Stipkovits, R. Rosengarten and J. Frey Report EUR 18756 EN EUROPEAN COMMISSION Edith CRESSON, Member of the Commission ■k responsible for research, innovation, education, Itraining and youth DG XII/B.1 — RTD actions: Cooperation with non- member countries and international organisations — |*,< European Economic Area, COST, Eureka and international organisations Contact: Ms F. Coudert Address: European Commission, rue de la Lot 200 (SDME 1/39) B-1049 Brussels — Tel. (32-2) 29 65507; fax (32-2) 29 64289 Cover illustration: Healthy Valais black-nosed sheep in an Alpine meadow. (Courtesy of Dr Marco Giacometti, Institute of Animal Pathology, University of Berne, Switzerland) COST COST s 4 European Cooperation on Scientific and European Commission Technical Research COST 826 Agriculture and biotechnology Mycoplasmas of ruminants: pathogenicity, diagnostics, epidemiology and molecular genetics Volume 3 Edited by: László Stipkovits Veterinary Medical Research Institute Budapest, Hungary Renate Rosengarten University of Veterinary Medicine Vienna, Austria Joachim Frey Institute for Veterinary Bacteriology, University of Berne Berne, Switzerland Directorate-General Science, Research and Development 1999 EUR 18756 EN A great deal of additional information on the European Union is available on the Internet. It can be accessed through the Europa server (http://europa.eu.int). Cataloguing data can be found at the end of this publication. Luxembourg: Office for Official Publications of the European Communities, 1999 ISBN 92-828-5222-9 © European Communities, 1999 Reproduction is authorised provided the source is acknowledged. Printed in Belgium PRINTED ON WHITE CHLORINE-FREE PAPER PREFACE The industrialised countries suffered during the last decade from a marked increase of re-emerging and new emerging infectious diseases which affected humans and animals. Among those, mycoplasmal diseases of animals are of particular concern. They constitute a threat for animal health and livestock production. The control and elimination of these infectious diseases constitute a challenge to veterinarians, officials of veterinary services, researchers and industry. The aim of COST Action 826 „Ruminants' Mycoplasmoses" is to provide a network for the exchange of scientific and technical knowledge, and for the promotion and co-ordination of research and development in the field of mycoplasmal diseases of ruminants. This Action is expected to rapidly provide the necessary elements for the elaboration of strategies for the prevention and eradication of ruminants' mycoplasmoses. Recent discoveries on the phylogenetic relationships among the different Mycoplasma species involved in ruminants' mycoplasmoses revealed the difficulty to clearly differentiate by genetic methods some of these species. It manifested the urge for more basic research to be done with this important group of mycoplasmas. This report is the result of the „Third Workshop on Mycoplasmas of Ruminants: Pathogenicity, Epidemiology, and Molecular Genetics" of COST Action 826 held in Budapest, Hungary, from May 5-8 1998. It resumes the recent outcomes in basic and applied research in the field of mycoplasmas of ruminants and the newest results on the prevalence of ruminants' mycoplasmoses in the different European regions. The Workshop was organised by the Veterinary Medical Research Institute of Hungarian Academy of Sciences at Hotel Agro located on the Sváb Hill, overlooking the Danube and the city of Budapest. It gave to all participants the unique chance to attend a most dynamic workshop who brought together 80 veterinarians, officials, researchers and industrials from many European countries and from other continents working in the different disciplines of ruminants' mycoplasmoses. The workshop showed which directions the development of diagnostic methods and preventive concepts will take in the future. I would like to thank Prof. Laszlo Stipkovits and his collaborators for the excellent organisation of the Workshop and for providing the participants not only an outstanding scientific programme but also giving them the opportunity to get an insight in Hungarian live and culture. I would like to thank Prof. Renate Rosengarten, Vienna for her precious input during the preparations and our COST Secretary Dr. Françoise Coudert, Brussels for her help in the realisation of this Workshop. Particular thanks to the sponsors: The National Committee for Technological Development, Budapest; The Hungarian Academy of Sciences, Budapest; Pfizer A.H.S.A.,; Bayer A.G., Leverkusen, Germany; Novartis Animal Health GmbH, Kundl, Austria; Laboratorios Calier S.A., Barcelona, Spain; and Elanco Animal Science Research, Eli Lilly and Company Ltd, Basingstoke, UK. The sponsors manifested with their financial support their interest in our research activities and made possible the organisation of this most productive Workshop. Berne, August 4 1998 Joachim Frey Chairman, COST 826 CONTENTS Overviews Antigenic variation in mycoplasmas: past, present and future Cittì, C, Poumarat, F, Le Grand, D, Bergonier, D., Beier, T., Brank, M., Glew, M.D., Rosengarten, R 1 CBPP andCCPP,historicalreview and actual threat for Europeancattle Thiaucourt,F.5 The role ofruminantmycoplasmas in systemic infections Rosengarten,R.,Cittì,C14 The impact of mycoplasma infections in respiratory diseases of cattle in Europe Reeve­Johnson, L 18 Impactofmycoplasma mastitis in cattle Ball,H.J:32 MolecularGenetics DiagnosisofCBPPby PCR and laser induced fluorescence Persson,Α.,Wahlberg, J., Johansson, K.E 36 DNA repair genes uvrCasgenetictargetsfor discrimination of closely related mycoplasmas Frey, J., Subramaniam, S.,Bergonier,D.,Nicolet, J40 Determination of genome size andrestriction pattern polymorphism of Mycoplasma agalactiae and Mycoplasma bovis by pulsed field gel electrophoresis Tola, S., Idini, G., Rocchigiani, A.M., Manunta, D., Rocca, S., Angioi, P.P., Cocco, M., Leori, G. 44 CharacterizationofhspSO and hsp70 genes of Mycoplasmas Scherm,B.,Niewint,J., Gerlach, G.F., Thiaucourt, F., Runge, M.47 OccurenceofvspAsequences in Mycoplasma bovis field isolates Poumarat,F.,Cittì,C, Legrand, D., Solsona, M., Rosengarten, R51 Intraspecific variation in the 16S rRNA gene sequences of Mycoplasma agalactiae strains Heldtander, M., Bölske, G., Johansson, K.E 52 Production and sequence analysisofaMycoplasmacapricolum subsp. capripneumoniae (F38­Biotype) recombinantexpression library Williamson, H.S., March, J.B.,56Biochemistry, physiology and antigenicity Variable surface proteins of Mycoplasma bovis. A role in cytoadhesion Sachse, K, Grajetzki, Ch., Heller, M., Yogev, D 60 Variable surface components inruminantandavian mycoplasmas: a comparative analysis Giew, M.D., Cittì, C, Bergonier, D., Rosengarten, R61 Antigenic variability of Mycoplasma agalactiae surface proteins Bergonier, D., Marenda, M., Citti, O, Glew, M.D., Fleury, B., Blanc, M.C., Sagne, E., Manesse, M., Costes, P., Berthelot, X., Rosengarten, R., Poumarat, F 65 Analysisofthecapsular polysaccharide of Mycoplasma mycoides subsp. mycoidesSCthecausal agent of CBPP: Purification composition and its role ininfectionandimmunity March,J.B.,Jones, G.E., Hitchen, P., Morris, H.R., Dell, A 69 Genetic an immunogenic characterisation of a 62KDalipoprotein (P62) in Mycoplasma mycoides subsp. mycoides LC and Mycoplasmamycoides subsp.capri Monnerat, M.P., Frey, J., Thiaucourt, F., Poveda, J.B., Nicolet, J 73 Development of coloured Mycoplasma mycoides coloniesonsolidmedia as an aid to rapid diagnosis Bashiruddin, J.B., Windsor, G.D., Regalia, J., De Santis, Ρ 77 Temperature tolerance of Mycoplasmas in the Mycoplasmamycoides cluster Bölske, G., Schantz, Κ 81 Humoralimmuneresponsein ewes experimentally infected with Mycoplasma mycoidessubsp.mycoidesSC isolated from cattle and sheep Gonçalves,R.,Belo,Α.,Goulao, J.V., Ferreira­Dias, G. 85 Isolationofthe60KD­heatshock protein ofMycoplasmaarthritidis and Mycoplasmabovisanddetection of heat shockproteinspecific antibodies in bovine sera Wedde, Α., Gerlach, G.F., Scherm, Β., Runge, M. 90 Diagnostics and field studies of M.agalactiaeand M. bovis Prevalence of Mycoplasmas in respiratory disease in cattle in Britain Nicholas, R., Ayling, R., Miles, R 94 Specific detection ofMycoplasmaagalactiae by PCR and nested­PCR based on the amplificationoftheuvrCgene Fleury, B., Bergonier,D.,Sagne,E.,Manesse, M., Berthelot, X., Nicolet, J., Frey, J 97 VI Isolation and characterization of Mycoplasma bovis from a calf pneumonia outbreak In Greece Filiousis, G., Papanikolaou, E., Petridou, E., Arsenakis, M., Sarris, K 101 Approaches to the development of selective media for Mycoplasmaagalactiae and Mycoplasma bovis Ozean, S., Rice, P., Al-Sheikhly, D., Suchak, Α., Patel, P., Miles, R 105 Isolation and characterization of Mycoplasma agalactiae strainsisolated in Greece Petridou, E., Papanikolaou, E., Filiousis, G., Arsenakis, M., Sarris, Κ 109 Enzootic Mycoplasma bovis infection in a Dairy Herd Stipkovits, L, Ripley, R., Varga.J 113 Identification and antibioticsusceptibilityofmycoplasmas associatedwith contagious agalactia inSicily Loria, G.R., Pirano, C,Worth,D.R.,Caracappa, S., Nicholas, R.A.J116 The control and eradicationschemeofcontagious agalactia in France:'The role of CNEVA Sophia Antipolis Pepin, M., Lambert, M., Sanchis, R., Abadie, G., Dufour, P., Russo, P., Vitu, C. ...119 Outbreaks of contagious agalactiae in Israel: Clinical and epizooticai aspects Rapoport, E., Flitman-Tene, R., Yogev, D., Levisohn, S 120 Mycoplasmas in a problem flock of sheep withcontagious agalactia De Santis, P., Bashruddin, J.B., Tittarelli, M.,Vissagio,M.C, Gianvincenzo,D...124 A new approach for Mycoplasma bovis serodiagnosis LeGrand, D., Citti, O, Bézille, P., Rosengarten, R., Poumarat, F. 127 Pathology of induced Mycoplasma bovis calf pneumonia inexperimentally vaccinated animals Bryson, D.G., Ball, H.J., Brice, Ν., Forster, F., Pollock, D.S. 128 Isolation of Mycoplasma bovis from the abomasalcontentsofanaborted bovine foetus Byrne, W., Brennan, P., McCormack, M., Ball, H.J. 133 Enhancement of Mycoplasma bovisdetectioninmilksamples by antigen capture prior to PCR Hotzel, H., Heller, M., Sachse, Κ. 136 Comparisonoffoursamplingprocedures for identification of respiratory mycoplasmasincattle Thomas, Α.,Schreiber,P.,Pirson, V, Mainil, J., Linden, A 137 Examination ofantibodyresponseof cows and theircalvestoMycoplasma bovis by Western blot Fabian, K., Tenk, M., Stipkovits, L141 VII Epidemiological study of mycoplasma infections in calves Zendulkova, D., Lany, P., Pospisil, Z., Skrivanek, M 144 Use of Elisa „CIVTEST Mycoplasma agalactiae„forthedetection of specific IgG antobodies in sheep vaccinated withAG1strain Muhedini, P., Mati, D., Santini, F.G 148 Diagnostics and fieldstudiesofmycoplasmas of the M. mycoides cluster Validation of a competitiveElisaforcontagious Bovine Pleuropneumonia Thiaucourt, F., Geiger,R.,Nicolet,J.,Pini, Α., Prozesky, L., Regalla, J., Roger, F., Yaya, Α. 152 A field testfordetectingantibodies to Mycoplasma mycoides subsp.mycoides SC typeusingthelatexslideagglutination test Ayling, R.,Regalia,J.,Nicholas, R155 Studiesontheimmunologicaldiversity of type, vaccine and wildstrainsof Mycoplasmamycoidessubsp.mycoides SC variant March, J.B., Jones, G.E., Williamson, H.S., Amanfu, W 159 Renal lesions in cattle with contagious BovinePleuropneumonia: an immunohistochemical study Scanziani, E., Grieco, V, Boldini, M., Monaci, O,Paltrinieri,S.,Luini, M. 163 Evaluation of colorimetrie PCR for the diagnosis of CBPP Botelho, Α., Prata, Α., Bashiruddin, J., Regalla, J 166 Modification of the complement fixationtest(CFT)for CBPP Tittarelli, M., Conte, A.M., Rodomonti,D.,DeSantis, Ρ170 Application of PCR for the identificationofruminantMycoplasmas isolated in Belgium D'hooghe, I., Pohl, P., De Feyter, E., Peeters, J 173 Investigation on the current statusofcaprinemycoplasmosis in the Eastern Hills of Nepal Spergser, J., JHA, V.C., Hotzel, H.,Beier,T.175 Further epizootiological and microbiological observations on an outbreak of ovine infectious keratoconjunctivitis in Croatia Naglic, T., Hajsig, D., Seol, B., Busch, Κ 178 Evaluation of bovinewholebloodneutrophil viability upon interactionwith Mycoplasma mycoidessubsp.mycoidesSC Ferronha, M.H., Correia,I.,Trindade,Η.,Carvalho, Η., Amaro, A 182 Experimental transmissioninsheepandgoats of Mycoplysmamycoides subsp. mycoides LC from bovineorigin Machado, M., Ferreira, H., Ramos, M.A.,Brandão, E., Tavares, Α., Cardoso,M... 186 Author index 190 VIII