Detection of Mycoplasma bovis Infection in Cattle Mammary Tissue by Immunofluorescence and qRT-PCR Methods
Yıl 2019,
, 110 - 115, 30.06.2019
Serdar Altun
,
Selçuk Özdemir
Öz
Mastitis in cattle
causes important economic losses in all over the world and Mycoplasma bovis is one of the important agent among pathogens
shown to be responsible for this disease. The aim of this study was to
determine the presence and
prevalence of Mycoplasma bovis among
the pathological agents implicated in mastitis by immunofluorescence staining
and qRT-PCR methods in cattle mammary tissue. For this aim, 120 mammary samples
with or without macroscopic lesions as this agent causes subclinical mastitis,
were collected. Mastitis was
diagnosed in 78 of 120 cases. Chronic mastitis cases in 56 and, acute mastitis
in 22 cases were histopathologically diagnosed.
Fluorescent positivity was determined 28/120 according to the results of
immunofluorescence staining for all sections. Immunpositive signs of Mycoplasma bovis were commonly observed
in samples with chronic mastitis. These results were confirmed by qRT-PCR. According
to these results the presence of Mycoplasma
bovis agent that an important threat to herd health is 23,3%.
Kaynakça
- Giovannini S, Zanoni MG, Salogni C, Cinotti S, Alborali GL. Mycoplasma bovis infection in respiratory disease of dairy calves less than one month age. Resaerch in Veterinary Sciences, 2013; 95: 576-579.
- Amram E, Freed M, Khateb N, Mükula I, Blum S, Spergser J, Sharir B, Ozeri R, Harrus S, Lysnyansky I. Multiple locus variable number tandem repeat analysis of Mycoplasma bovis isolated from local and imported cattle, The Veterianary Journal, 2013; 197: 286-290.
- Tamada Y, Ohura K, Kato K, Kimura K, Maeda T. Mycoplasma bovis infection in calves with ear drops. Journal of the Hokkaido Veterinary Medical Association. 2002; 46: 308-311.
- Adamu JY, Wawegama NK, Browning GF, Markham PF. Membrane proteins of Mycoplasma bovis and their role in pathogenesis. Research in Veterinary Science, 2013; 95: 321-325.
- Aebi M, Bodmer M, Frey J, Pilo P. Herd-specific strains of Mycoplasma bovis in outbreaks of mycoplasmal mastitis and pneumonia. Veterinary Microbiology, 2012; 157: 363–368.
- Nicholas RAJ, Ayling RD, Stipkovits LP. An experimental vaccine for calf pneumonia caused by Mycoplasma bovis: clinical, cultural, serological and pathological findings, Vaccine, 2002; 20: 3569-3575.
- Karahan M, Kalin R, Atil E, Çetinkaya B. Detection of Mycoplasma bovis in cattle with mastitis and respiratory problems in eastern Turkey. Veterinary Record, 2010; 166: 827-829.
- Radelli E, Luini M, Loria GR, Nicholas RAJ, Scanziani E. Bacteriological, serological, pathological and immunohistochemical studies of Mycoplasma bovis respiratory infection in veal calves adult cattle at slaughter. Research in Veterinary Sciences, 2008; 85: 282-290.
- Wooley L K, Fell S, Gonsalves JR, Walker MJ, Djordjevic SP, Jenkins C, Eamens GJ. Evaluation of clinical, histological and immunological changes and qPCR detection of Mycoplasma hyopneumoniae in tissue during the eraly stages of mycoplasmal pneumonia in pigs after experimental challenge with two field isolates. Veterinary Microbiology, 2012; 161: 186-195.
- Grand DL, Calavas D, Brank M, Cıttı C, Rosengarten R, Be´zılle P, Poumarat F. Serological prevalence of Mycoplasma bovis infection in suckling beef cattle in France Veterinary Record, 2002; 150: 268-273.
- Ayling R, Nicholas R, Hogg R, Wessels J, Scholes S, Byrne W, Hill M, Moriarty J, Brien T. Mycoplasma bovis isolated from brain tissue of calves, 2005; 156: 391-392.
- Soehnlen MK, Kunze ME, Karunathilake E, Henwood BH, Karyawasam S, Wolfgang DR, Jayarao BM. İn vitro antimicrobial inhibition of Mycoplasma bovis isolates submitted to the Pennsylvania Animal Diagnostic Laboratory using flow cytometry and broth microdilution method. Journal of Veterinary Daignostic İnvestigation, 2011, 23(3); 547-551.
- Erdağ O, Erdoğan G, Türkaslan J, Gürel A. Buzağı ve dana pneumonilerinde Mycoplasma ve bakteriyel etkenlerin izolasyon, identifikasyon ve antibiyotik duyarlılıkları. Animal Information, 1995; 112: 115-119.
- Schott C, Cai H, Parker L, Bateman K G, Caswell L. Hydrogen peroxide produciton and free radical mediated cell stress in Mycoplasma bovis pneumonia. Journal of Compharative Pathology, 2013; 1-11.
- Kleinschmidt S, Spergser J, Rosengarten R, Hewicker Trautwein M. Long-term survival of Mycoplasma bovis in necrotic lessions and in phagocyctc cells as demonstrated by transmission and immunogold electron microscopy in lung tissue from experimantally infected calves. Veterinary Microbiology, 2013; 162: 949-953.
- Stipkovits L, Ripley P, Varga J, Pálfi V. Clinical study of the disease of calves associated with Mycoplasma bovis infection. Acta Veterinaria Hungarica, 2000; 48(4):387-95.
- Rosetti BC, Frey J, Pilo P. Direct detection of Mycoplasma bovis in milk and tissue samples by real-time PCR. Moleculer and Cellular Probes, 2010; 24: 321-323.
- Haapalaa V, Pohjanvirtab T, Vähänikkiläb N, Halkilahtic J, Simonend H, Pelkonenb S, Soveria T, Simojokia H, Autiob T. Semen as a source of Mycoplasma bovis mastitis in dairy herds. Veterinary Microbiology, 2018; 216: 60–66.
- Fan Q, Xie Z, Xie Z, Xie L, Huang J, Zhang Y, Zeng T, Zhang M, Wan S, Luo S, Liu J, Deng X. Development of duplex fluorescence-based loop-mediated isothermal amplification assay for detection of Mycoplasma bovis and bovine herpes virus 1. J Virol Methods. 2018; 261:132-138.
Sığır Meme Dokularında Mycoplasma bovis Enfeksiyonunun İmmunofloresan ve qRT-PCR Yöntemleriyle Araştırılması
Yıl 2019,
, 110 - 115, 30.06.2019
Serdar Altun
,
Selçuk Özdemir
Öz
Sığırlarda Mastitis
tüm dünyada önemli ekonomik kayıplara neden olmaktadır. Mycoplasma
bovis ise bu hastalıktan sorumlu olarak gösterilen patojenler arasında en
önemli bakteriyel etkenlerden biridir. Bu çalışmanın amacı, sığır meme
dokusunda oluşan mastitisde rol oynayan patolojik ajanlar arasında Mycoplasma
bovis'in varlığı ve prevalansını immunofloresan boyama ve qRT-PCR yöntemleriyle
ve saptamaktır. Bu amaçla, etkenin
subklinik mastitise yol açtığı düşünülerek, makroskobik olarak lezyonlu ya da
lezyonsuz olunmasına bakılmaksızın 120 adet meme örneği toplandı. Toplanan 120
örneğin 56 kronik ve 22 akut olmak üzere toplam 78 örnekte histopatolojik
olarak mastitis tanısı kondu. Tüm
kesitlere uygulanan immünofloresan boyama sonuçlarına göre floresan pozitifliği
28/120 olarak belirlendi. Mycoplasma
bovis immünopozitiflikleri daha yoğun olarak kronik mastitisli örneklerde
gözlendi. Bu sonuçlar örnklere uygulanan qRT-PCR sonuçları ile uyumludur. Bu
sonuçlara göre, Erzurum'da sürü sağlığı için önemli bir tehdit olan Mycoplasma bovis etken varlığı %23,3
oranında tespit edilmiştir.
Kaynakça
- Giovannini S, Zanoni MG, Salogni C, Cinotti S, Alborali GL. Mycoplasma bovis infection in respiratory disease of dairy calves less than one month age. Resaerch in Veterinary Sciences, 2013; 95: 576-579.
- Amram E, Freed M, Khateb N, Mükula I, Blum S, Spergser J, Sharir B, Ozeri R, Harrus S, Lysnyansky I. Multiple locus variable number tandem repeat analysis of Mycoplasma bovis isolated from local and imported cattle, The Veterianary Journal, 2013; 197: 286-290.
- Tamada Y, Ohura K, Kato K, Kimura K, Maeda T. Mycoplasma bovis infection in calves with ear drops. Journal of the Hokkaido Veterinary Medical Association. 2002; 46: 308-311.
- Adamu JY, Wawegama NK, Browning GF, Markham PF. Membrane proteins of Mycoplasma bovis and their role in pathogenesis. Research in Veterinary Science, 2013; 95: 321-325.
- Aebi M, Bodmer M, Frey J, Pilo P. Herd-specific strains of Mycoplasma bovis in outbreaks of mycoplasmal mastitis and pneumonia. Veterinary Microbiology, 2012; 157: 363–368.
- Nicholas RAJ, Ayling RD, Stipkovits LP. An experimental vaccine for calf pneumonia caused by Mycoplasma bovis: clinical, cultural, serological and pathological findings, Vaccine, 2002; 20: 3569-3575.
- Karahan M, Kalin R, Atil E, Çetinkaya B. Detection of Mycoplasma bovis in cattle with mastitis and respiratory problems in eastern Turkey. Veterinary Record, 2010; 166: 827-829.
- Radelli E, Luini M, Loria GR, Nicholas RAJ, Scanziani E. Bacteriological, serological, pathological and immunohistochemical studies of Mycoplasma bovis respiratory infection in veal calves adult cattle at slaughter. Research in Veterinary Sciences, 2008; 85: 282-290.
- Wooley L K, Fell S, Gonsalves JR, Walker MJ, Djordjevic SP, Jenkins C, Eamens GJ. Evaluation of clinical, histological and immunological changes and qPCR detection of Mycoplasma hyopneumoniae in tissue during the eraly stages of mycoplasmal pneumonia in pigs after experimental challenge with two field isolates. Veterinary Microbiology, 2012; 161: 186-195.
- Grand DL, Calavas D, Brank M, Cıttı C, Rosengarten R, Be´zılle P, Poumarat F. Serological prevalence of Mycoplasma bovis infection in suckling beef cattle in France Veterinary Record, 2002; 150: 268-273.
- Ayling R, Nicholas R, Hogg R, Wessels J, Scholes S, Byrne W, Hill M, Moriarty J, Brien T. Mycoplasma bovis isolated from brain tissue of calves, 2005; 156: 391-392.
- Soehnlen MK, Kunze ME, Karunathilake E, Henwood BH, Karyawasam S, Wolfgang DR, Jayarao BM. İn vitro antimicrobial inhibition of Mycoplasma bovis isolates submitted to the Pennsylvania Animal Diagnostic Laboratory using flow cytometry and broth microdilution method. Journal of Veterinary Daignostic İnvestigation, 2011, 23(3); 547-551.
- Erdağ O, Erdoğan G, Türkaslan J, Gürel A. Buzağı ve dana pneumonilerinde Mycoplasma ve bakteriyel etkenlerin izolasyon, identifikasyon ve antibiyotik duyarlılıkları. Animal Information, 1995; 112: 115-119.
- Schott C, Cai H, Parker L, Bateman K G, Caswell L. Hydrogen peroxide produciton and free radical mediated cell stress in Mycoplasma bovis pneumonia. Journal of Compharative Pathology, 2013; 1-11.
- Kleinschmidt S, Spergser J, Rosengarten R, Hewicker Trautwein M. Long-term survival of Mycoplasma bovis in necrotic lessions and in phagocyctc cells as demonstrated by transmission and immunogold electron microscopy in lung tissue from experimantally infected calves. Veterinary Microbiology, 2013; 162: 949-953.
- Stipkovits L, Ripley P, Varga J, Pálfi V. Clinical study of the disease of calves associated with Mycoplasma bovis infection. Acta Veterinaria Hungarica, 2000; 48(4):387-95.
- Rosetti BC, Frey J, Pilo P. Direct detection of Mycoplasma bovis in milk and tissue samples by real-time PCR. Moleculer and Cellular Probes, 2010; 24: 321-323.
- Haapalaa V, Pohjanvirtab T, Vähänikkiläb N, Halkilahtic J, Simonend H, Pelkonenb S, Soveria T, Simojokia H, Autiob T. Semen as a source of Mycoplasma bovis mastitis in dairy herds. Veterinary Microbiology, 2018; 216: 60–66.
- Fan Q, Xie Z, Xie Z, Xie L, Huang J, Zhang Y, Zeng T, Zhang M, Wan S, Luo S, Liu J, Deng X. Development of duplex fluorescence-based loop-mediated isothermal amplification assay for detection of Mycoplasma bovis and bovine herpes virus 1. J Virol Methods. 2018; 261:132-138.