Direct detection of Mycoplasma bovis in milk and tissue samples by real-time PCR

Rossetti, Bigna C.; Frey, Joachim; Pilo, Paola

doi:10.1016/j.mcp.2010.05.001

Cited by 45 publications

(47 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In order to identify M. bovis positive animals, milk, nasal swabs and organs were tested for M. bovis by a direct real-time PCR as previously described by Rossetti et al [25]. Briefly, lysates from samples were prepared as DNA template [25,26].…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, lysates from samples were prepared as DNA template [25,26]. Fifty μl of milk sediment was added to 200 μl of lysis buffer (0.1 M Tris–HCl, pH 8.5, 0.05% Tween 20, 0.24 mg/ml proteinase K) and swabs taken from gross lesions in organs and nasal swabs were dipped for 1 min in 500 μl of lysis buffer.…”

Section: Methodsmentioning

confidence: 99%

“…All lysates were incubated for 1 h at 60°C and 15 min at 95°C as a denaturation step. To identify M. bovis in samples, a real-time PCR specific for M. bovis and targeting the uvrC gene was further performed using a Ct cutoff value of 40 [25,26]. This method detects approximately 2,000 colony-forming units in the sediment of 1 ml of milk and does not cross react with M. agalactiae, M. capricolum subsp.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Mycoplasma bovis infections in Swiss dairy cattle: a clinical investigation

et al. 2015

Self Cite

View full text Add to dashboard Cite

Mycoplasma bovis causes mastitis in dairy cows and is associated with pneumonia and polyarthritis in cattle. The present investigation included a retrospective case–control study to identify potential herd-level risk factors for M. bovis associated disease, and a prospective cohort study to evaluate the course of clinical disease in M. bovis infected dairy cattle herds in Switzerland. Eighteen herds with confirmed M. bovis cases were visited twice within an average interval of 75 d. One control herd with no history of clinical mycoplasmosis, matched for herd size, was randomly selected within a 10 km range for each case herd. Animal health data, production data, information on milking and feeding-management, housing and presence of potential stress- factors were collected. Composite quarter milk samples were aseptically collected from all lactating cows and 5% of all animals within each herd were sampled by nasal swabs. Organ samples of culled diseased cows were collected when logistically possible. All samples were analyzed by real-time polymerase chain reaction (PCR). In case herds, incidence risk of pneumonia, arthritis and clinical mastitis prior to the first visit and incidence rates of clinical mastitis and clinical pneumonia between the two visits was estimated. Logistic regression was used to identify potential herd-level risk factors for M. bovis infection. In case herds, incidence risk of M. bovis mastitis prior to the first visit ranged from 2 to 15%, whereas 2 to 35% of the cows suffered from clinical pneumonia within the 12 months prior to the first herd visit. The incidence rates of mycoplasmal mastitis and clinical pneumonia between the two herd visits were low in case herds (0–0.1 per animal year at risk and 0.1-0.6 per animal year at risk, respectively). In the retrospective-case-control study high mean milk production, appropriate stimulation until milk-let-down, fore-stripping, animal movements (cattle shows and trade), presence of stress-factors, and use of a specific brand of milking equipment, were identified as potential herd-level risk factors. The prospective cohort study revealed a decreased incidence of clinical disease within three months and prolonged colonization of the nasal cavity by M. bovis in young stock.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Mycoplasma bovis infections in Swiss dairy cattle: a clinical investigation

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

“…are mollicutes, cell wall-less, slow growing organisms that require special culture media and growth conditions. For Mycoplasma spp., as for Prototheca spp., many molecular studies focus on detection and species identification, especially because molecular diagnostics are faster than culture [8, 81, 164, 165]. As for other bacterial mastitis pathogens, outbreak investigations and routes of transmission are the main subjects of molecular epidemiological studies.…”

Section: Other Mastitis Pathogens: Prototheca and Mycoplasmamentioning

confidence: 99%

Molecular Epidemiology of Mastitis Pathogens of Dairy Cattle and Comparative Relevance to Humans

Zadoks

Middleton

McDougall³

et al. 2011

J Mammary Gland Biol Neoplasia

358

291

View full text Add to dashboard Cite

Mastitis, inflammation of the mammary gland, can be caused by a wide range of organisms, including gram-negative and gram-positive bacteria, mycoplasmas and algae. Many microbial species that are common causes of bovine mastitis, such as Escherichia coli, Klebsiella pneumoniae, Streptococcus agalactiae and Staphylococcus aureus also occur as commensals or pathogens of humans whereas other causative species, such as Streptococcus uberis, Streptococcus dysgalactiae subsp. dysgalactiae or Staphylococcus chromogenes, are almost exclusively found in animals. A wide range of molecular typing methods have been used in the past two decades to investigate the epidemiology of bovine mastitis at the subspecies level. These include comparative typing methods that are based on electrophoretic banding patterns, library typing methods that are based on the sequence of selected genes, virulence gene arrays and whole genome sequencing projects. The strain distribution of mastitis pathogens has been investigated within individual animals and across animals, herds, countries and host species, with consideration of the mammary gland, other animal or human body sites, and environmental sources. Molecular epidemiological studies have contributed considerably to our understanding of sources, transmission routes, and prognosis for many bovine mastitis pathogens and to our understanding of mechanisms of host-adaptation and disease causation. In this review, we summarize knowledge gleaned from two decades of molecular epidemiological studies of mastitis pathogens in dairy cattle and discuss aspects of comparative relevance to human medicine.

show abstract

“…The study of the sequences of the uvrC encoding gene sequence may lead to understanding of the phylogenic relations of different strains of M. bovis species. Rossetti used a novel designed pair of primers for the amplification of the uvrC gene by Real-time-PCR as the bio marker of M. bovis strains (Rossetti et al, 2010). Because of the important role of M. bovis in clinical mastitis in dairy herds, this study was designed to isolate and characterize M. bovis strains originated from different clinical mastitis cases from all over Iran, using two traditional culture-biochemical methods in comparison to highly sensitive molecular tools such as the PCR method.…”

Section: Introductionmentioning

confidence: 99%

Isolation, identification and molecular characterization of Mycoplasma bovis in mastitic dairy cattle by PCR and culture methods

Imandar

Pourbakhsh

Jamshidian

et al. 2018

J Hellenic Vet Med Soc

View full text Add to dashboard Cite

Mycoplasma bovis is well known as one of the major causative agents of mastitis in dairy cattle herds. The aim of this study was the identification of Mycoplasma bovis strains by PCR and traditional culture methods from a total number of 328 milk samples collected from cows with clinical mastitis symptoms from all over Iran. First step cultures in PPLO broth and agar showed 58 samples (17.69%) as positive. Out of 328 samples, 97 samples (29.57%) were positive for Mycoplasma genus according to the amplification of the 16SrRNA gene performed by PCR and from them, 31 (31.97%) samples were positive by PCR on the P48 gene. The purified P48 positive PCR products were sequenced and results were compared to M. bovis reference strain PG45 (CP002188). A phylogenic tree was created using Neighbor-joining method in MEGA6 software. The studied strain IB220 showed 100% identity with the reference strain of M. bovis and followed the same phylogenetic roots while studied strain IB216 showed 99.7% homology with the reference strain. Twelve selected geographical isolated strains were subjected to Gene Bank under accession numbers of KX772789 to KX772800. This is the first study of the molecular characterization of Mycoplasma bovis in dairy cattle with clinical mastitis from Iran.

show abstract

Direct detection of Mycoplasma bovis in milk and tissue samples by real-time PCR

Cited by 45 publications

References 15 publications

Mycoplasma bovis infections in Swiss dairy cattle: a clinical investigation

Mycoplasma bovis infections in Swiss dairy cattle: a clinical investigation

Molecular Epidemiology of Mastitis Pathogens of Dairy Cattle and Comparative Relevance to Humans

Isolation, identification and molecular characterization of Mycoplasma bovis in mastitic dairy cattle by PCR and culture methods

Contact Info

Product

Resources

About