аЯрЁБс>ўџ 46ўџџџ5џџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџьЅС5@№Пє(bjbjЯ2Я2 .*­X­Xв џџџџџџˆ2222222F    4> F ЖVVVVV111œ ž ž ž ž ž ž г R%цž 92F 11F F ž 22VVлз к к к F R2V2Vœ к F œ к .к  22 VJ ЫЫm‚Щ ˜  œ э 0  Ђ .  FF2222 2 ”1Њл0к єџG111ž ž FFФ а FF Incidence of Mycoplasma bovis in nasal cavities of calves and confirmation of mycoplasmal pneumonia in Croatian calves K. Matanovi1, B. `eol1*, S. Meki1, H. J. Ball2, M. Martinec3, Z. Kutla a3, D. Kova i3 1Department of Microbiology and Infectious Diseases, University of Zagreb, Faculty of Veterinary Medicine, Heinzelova 55, 10000 Zagreb, Croatia 2Veterinary Research Laboratories, Stoney Road, Stormont, Belfast, BT4 3SD, Northern Ireland 3Veterinary Station Remetinec, Brezovi ka 105, 10257 Hrvatski Leskovac, Croatia *Corresponding author Abstract Mycoplasma bovis was first isolated from milk of cows with mastitis in USA. Throughout the years, its significance has also been proved in arthritis, pneumonia, otitis and conjunctivitis in calves. In recent years, M. bovis was frequently isolated from nasal cavities and/or pneumonic lungs of calves in many countries in North America, Europe, and Asia. In our previous work we confirmed that M. bovis is a significant and widespread cause of bovine mastitis in Croatia, however there are no data about its role in respiratory diseases of calves. The aim of this study was to investigate the prevalence of nasal carriage of M. bovis and to confirm its role in pneumonia of calves in Croatia. Four farms on which calves from various sources were raised and had continuous problems with chronic pneumonia were selected for the study. M. bovis was suspected as a cause of pneumonia because there were no improvements after prolonged antibiotic therapy. Sixty six nasal swabs and 9 lung samples of calves from 4 farms were obtained and investigated for the presence of M. bovis using enrichment and capture ELISA, culture in Hayflick broth and culture on mycoplasma agar. 10 nasal swabs and all lung samples were also plated on Columbia agar with 5% sheep blood (Biorad, France) for routine bacteriological analysis. After isolation on solid agar and identification by ELISA, 20 isolates were randomly selected and tested by polymerase chain reaction specific. M. bovis was detected in 36 out of 66 (54.5%) nasal swabs and 3 out of 9 (30%) lung samples by enrichment and capture ELISA. All samples positive in ELISA yielded growth of mycoplasma colonies on mycoplasma agar plates. All 20 selected isolates that were previously identified by ELISA were confirmed as M. bovis by PCR. Out of 10 tested nasal swabs, 90% were positive for both M. bovis and Pasteurella multocida. P. multocida and Mannheimia haemolytica only were cultured from one lung sample each, while other 7 samples were negative for bacteria. This work confirmed our assumption that M. bovis is a frequent colonizer of nasal cavities of calves on Croatian farms. M. bovis was isolated for the first time from pneumonic lungs of calves in Croatia thus confirming its role as a cause of pneumonia in calves. However, as there is no routine testing of pneumonic lungs for the presence of mycoplasmas, this problem is often overlooked as more common pathogens such as P. multocida, M. haemolytica and Histophilus somni:ю№    4 6 8 N P R j l p r „ † Š ž   янявТА Т А“ А ƒqb q R q Rqhз#ќh8OŒ6CJaJmH sH hз#ќhN?CJaJmH sH "hз#ќhN?6CJH*aJmH sH hз#ќhN?6CJaJmH sH h3*6CJaJmH sH hз#ќh‹S^6CJaJmH sH "hз#ќh‹S^6CJH*aJmH sH hз#ќhЖ)y6CJaJmH sH hз#ќhЖ)ymH sH "hз#ќh'HЂ56CJaJmH sH hз#ќh'HЂ5CJaJmH sH ю№Ђ Є P Ф ~  % & / ђ(ѓ(є(їђчтййдЬЬЬСђМКgdЖ)y $dрa$gdЖ)y$a$gdЖ)ygdN? 7$8$H$gd‹S^gdŽTQ $dрa$gdЖ)ygd'HЂ$a$gd'HЂє(§  Ђ Є І  x Ф Ц ~ € – Є    & . / ?   юуюгПг­­г€гqbRCRhз#ќh'HЂCJaJmH sH hз#ќh'HЂ6CJaJmH sH hз#ќhЖ)yCJaJmH sH hз#ќhєIяCJaJmH sH h3*6CJaJmH sH hз#ќhŽTQ6CJaJmH sH hз#ќhN?6CJaJmH sH "hз#ќhN?6CJH*aJmH sH 'hз#ќh‹S^6CJaJmH nHsH tHhз#ќhЖ)y6CJaJmH sH hз#ќhŽTQmH sH "hз#ќhЖ)y6CJH*aJmH sH    ™ Ё Й К Л С  ЁЂЈщю(2pqsxЕСцяYZ[a„ПР!)QŒГЙжяпаСапяпапяпаСаСапяпаСаВапяпЃ”Ѓа…аvаЃаgаhз#ќhз#ќCJaJmH sH hз#ќhХ"ЅCJaJmH sH hз#ќhg{ЯCJaJmH sH hз#ќhN?CJaJmH sH hз#ќh .HCJaJmH sH hз#ќh*bЄCJaJmH sH hз#ќhћCJaJmH sH hз#ќh'HЂCJaJmH sH hз#ќh'HЂ6CJaJmH sH hз#ќh*bЄ6CJaJmH sH (жпрстущћќ   !%жкры"*,1>?@VётгУГУтЄтЄгЄт•т†тётУтw•w•whYhз#ќh'=ЇCJaJmH sH hз#ќhм4CJaJmH sH hз#ќhы]јCJaJmH sH hз#ќhћCJaJmH sH hз#ќh .HCJaJmH sH hз#ќhYtCJaJmH sH hз#ќh*bЄ6CJaJmH sH hз#ќh'HЂ6CJaJmH sH hз#ќh*bЄCJaJmH sH hз#ќh'HЂCJaJmH sH hз#ќhз#ќCJaJmH sH V[\]ch}~€‹šІЇЌј/017;€‡ghЌ­ЎИКёсбсТВЂ“ВбВ“ЂВ“„“u“„“ВбВ“ТсбсТfТсбсТhз#ќhІ?“CJaJmH sH hз#ќhы]јCJaJmH sH hз#ќh*bЄCJaJmH sH hз#ќh'HЂCJaJmH sH hз#ќh'=Ї6CJaJmH sH hз#ќh'HЂ6CJaJmH sH hз#ќhм4CJaJmH sH hз#ќh*bЄ6CJaJmH sH hз#ќhм46CJaJmH sH hз#ќhW9CJaJmH sH $КМШЭЮбо(A(t(u(v(|(€(‹(ђ(ѓ(є(япапРпОаЏŸŸЏ€Џqfhз#ќhЖ)ymH sH hз#ќhЖ)yCJaJmH sH hз#ќhћCJaJmH sH hз#ќh*bЄ6CJaJmH sH hз#ќh'HЂ6CJaJmH sH hз#ќh'HЂCJaJmH sH Uhз#ќhы]ј6CJaJmH sH hз#ќhІ?“CJaJmH sH hз#ќhІ?“6CJaJmH sH hз#ќhнl–6CJaJmH sH  are more easily isolated and attributed to pathologic conditions. There is an urgent need to stop the spreading of M. bovis by identifying heavily infected herds and establishing prevention and control measures in transport and herd market. ,1hА‚. АЦA!А‰"А‰#‰$‰%ААФАФ Фœ@@ёџ@ Ж)yNormalCJ_HaJmH sH tH DA@ђџЁD Default Paragraph FontRi@ѓџГR  Table Normalі4ж l4жaі (k@єџС(No Listв * џџџџwxбв(bП%&/а б д ˜@0€€˜@0€€˜@0€€˜@0€€˜@0€€˜@0€€˜@0€€˜@0€€˜@0€€˜@0€€˜@0€€˜@0€€˜@0€€˜@0€€   жVКє( є( є( №8№@ёџџџ€€€ї№’№№0№( № №№B №S №ПЫџ ?№џџs+ЪЄіХt+Ъ,u+Ъ4†Цv+Ъl"Хw+ЪЬOx+ЪЌ"Хy+Ъ”Сz+ЪlЖЩ{+ЪЄdЦ|+ЪФмХ}+Ъќ.~+ЪЌцЩ+Ъ,EЧ€+ЪьЦ+ЪtJХ‚+Ъ|Рƒ+ЪМЄЪ„+ЪLЙЧ…+ЪЦ†+Ъd'Ч‡+ЪOЧˆ+ЪLт‰+Ъ$бŠ+ЪФ_Ч‹+ЪšШŒ+Ъ\‰!+ЪtвŽ+ЪdnШ+ЪdGУ+Ъфр’RRZœœЅЎєєvvq€Œўўллјј#Ъ Ъ д      ›XaaЃЌООyy~†тт!))б б д    >*€urn:schemas-microsoft-com:office:smarttags €PostalCode€=*€urn:schemas-microsoft-com:office:smarttags €PlaceName€=*€urn:schemas-microsoft-com:office:smarttags €PlaceType€9*€urn:schemas-microsoft-com:office:smarttags€place€B*€urn:schemas-microsoft-com:office:smarttags€country-region€8*€urn:schemas-microsoft-com:office:smarttags€City€ …‹’šгмошєќ§д д ……д д џџKresimir Matanovicхйм43*N?W9ќ$9!? .HŽTQt1V‹S^YtЖ)yз:8OŒІ?“нl–'HЂ*bЄХ"Ѕ'=Ї0ЈšАg{Яр;лПMъы3эєIяы]јћз#ќџ@4о в     @ P@џџUnknownџџџџџџџџџџџџGю‡z €џTimes New Roman5€Symbol3&ю ‡z €џArial"qˆ№ФЉИыб†Иыб†žыᆂщщ щщ Љ#№‰‰ДДr4Ь Ь 3ƒq№HX)№џ?тџџџџџџџџџџџџџџџџџџџџџЖ)yџџHAn outbreak of Mycoplasma bovis mastitis on a dairy cows farm in CroatiaseolKresimir Matanovicўџр…ŸђљOhЋ‘+'Гй0Ь˜ є(8 T` | ˆ ”  ЌДМФтIAn outbreak of Mycoplasma bovis mastitis on a dairy cows farm in Croatiaon oseoltbreoleolNormalrKresimir Matanovicp2esMicrosoft Word 10.0@Œ))@МЏ‚Щ@ишP‚Щ@ишP‚Щщщ ўџеЭеœ.“—+,љЎ00 hp|„Œ” œЄЌД М тtЬ A IAn outbreak of Mycoplasma bovis mastitis on a dairy cows farm in Croatia Title ўџџџўџџџ !"#$%&ўџџџ()*+,-.ўџџџ§џџџ1ўџџџўџџџўџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџRoot Entryџџџџџџџџ РFhеm‚Щ3€1Tableџџџџџџџџ WordDocumentџџџџџџџџ.*SummaryInformation(џџџџDocumentSummaryInformation8џџџџџџџџџџџџ'CompObjџџџџџџџџџџџџjџџџџџџџџџџџџџџџџџџџџџџџџўџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџўџ џџџџ РFMicrosoft Word Document MSWordDocWord.Document.8є9ВqRoot Entryџџџџџџџџ РFа8 Ѕ‚Щ9Р1Tableџџџџџџџџ WordDocumentџџџџџџџџ.*SummaryInformation(џџџџ ўџџџўџџџ !"#$%&ўџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџ8§џџџўџџџўџџџўџџџ7џџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџўџџџ ўџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџ@є€ќ <d_AdHocReviewCycleID_EmailSubject _AuthorEmail_AuthorEmailDisplayNameАЉїmatanovic_abstractkmatanov@vef.hrKresimir MatanovicDocumentSummaryInformation8џџџџџџџџџџџџCompObjџџџџџџџџџџџџjџџџџџџџџџџџџџџџџџџџџџџџџўџ џџџџ РFMicrosoft Word Document MSWordDocWord.Document.8є9ВqўџеЭеœ.“—+,љЎDеЭеœ.“—+,љЎt0 hp|„Œ” œЄЌД М тtЬ A IAn outbreak of Mycoplasma bovis mastitis on a dairy cows farm in Croatia Title”