Use of a novel real-time PCR technique to monitor and quantitate Mycoplasma bovis infection in cattle herds with mastitis and respiratory disease

Sachse, Konrad; Salam, Hala Sayed Hassan; Diller, Roland; Schubert, Evelyn; Hoffmann, Bernd; Hotzel, Helmut

doi:10.1016/j.tvjl.2009.10.008

Cited by 77 publications

(82 citation statements)

References 17 publications

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“…In this method, mycoplasmal colonies are visible due to their ability to absorb dye (Malinowski & Kłossowska, 2002). Polymerase Chain Reaction (PCR) and its modification -real-time PCR (rt-PCR) are the techniques for identifying mycoplasmas from biological material (Sachse et al 2010;McAuliffe et al, 2005;Miles et al, 2004;Vasconcellos et al, 2000) but they have some limitations. Technique which is devoid of these limitations is denaturing gradient gel electrophoresis (DGGE) that allows differentiation of sixty seven mycoplasma species from one sample, including thirteen bovine pathogens in this M. bovis, M. dispar, M. bovirhinis and M. canis (McAuliffe et al, 2005).…”

Section: Diagnosismentioning

confidence: 99%

Bovine Respiratory Syndrome (BRD) Etiopathogenesis, Diagnosis and Control

Bednarek¹,

Szymańska-Czerwińska²,

Dudek³

2012

A Bird's-Eye View of Veterinary Medicine

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Section: Diagnosismentioning

confidence: 99%

Bovine Respiratory Syndrome (BRD) Etiopathogenesis, Diagnosis and Control

Bednarek¹,

Szymańska-Czerwińska²,

Dudek³

2012

A Bird's-Eye View of Veterinary Medicine

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“…Indirect transmission is possible through ingestion of contaminated milk, feed or water, or contact with contaminated bedding or fomites (Maunsell et al, 2011). Additional sources of infectious material are mucosal surface secretions from the teat canal or genital tract of adult cattle, as well as contaminated frozen semen , Nicholas and Ayling, 2003, Pitcher and Nicholas, 2005, Sachse et al, 2010.…”

Section: Reservoir and Transmissionmentioning

confidence: 99%

“…In regards to M. bovis pneumonia, trans-tracheal wash or BAL samples are preferable to URT samples such as nasal swabs as isolation of M. bovis from the URT of pneumonic animals is not well correlated with its presence in the LRT or with clinical disease (Allen et al, 1991, Thomas et al, 2002b. Although nasal or conjunctival swabs are easily obtained and provide information on the infection status of the herd (Sachse et al, 2010), BAL fluid or samples of lung tissue are probably the only reliable method to determine whether the lung of an individual is infected or not . Lung lesions, udder and joint or pleural fluid are also suitable and can all be easily collected from deceased animals (Nicholas and Baker, 1998).…”

Section: Specimen Selection and Submissionmentioning

confidence: 99%

“…Polymerase chain reaction systems amplify a particular DNA or RNA 'target sequence' enabling rapid detection of that sequence which may be specific for a particular organism of interest from culture (Sachse et al, 2010). Using species specific primer pairs, this process allows the detection and identification of an individual mycoplasma species (Justice- Allen et al, 2011) from either a submitted clinical sample or a cultured isolate.…”

Section: Speciation Of Culturable Isolatesmentioning

confidence: 99%

“…Because PCR systems amplify a particular DNA or RNA 'target sequence', they enable rapid detection of that sequence which ideally is specific for the organism of interest, viable or not, from clinical material (Sachse et al, 2010). As PCR systems amplify a 'target sequence', their application with slow growing or non-culturable organisms, in addition to samples collected from animals treated with antimicrobials or with samples that have been stored for some time or incorrectly (Maunsell et al, 2011, Pinnow et al, 2001.…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

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Mycoplasma bovis in Australian feeder cattle

Schibrowski¹

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Mycoplasma bovis can be a commensal bacterial inhabitant of the upper respiratory tract of healthy bovines. However, under certain circumstances and conditions that are incompletely understood, M. bovis can be associated with a number of clinical syndromes, one of which is bovine respiratory disease (BRD). In Australia, there is little information on the prevalence and epidemiology M. bovis in beef production systems. The overall aim of this thesis was to help close this knowledge gap.Commercially available serological diagnostic methods for the detection of M. bovis-specific antibody have not been validated in the Australian cattle population. Arguably, more importantly, however is the fact that diagnostic sensitivity (Se) and specificity (Sp) and / or analytical sensitivity (ASe) and specificity (ASp) estimates for commercially available M. bovis sero-diagnostic tests are not available. 28% (95% CI: 1% -92%). However, in terms of Sp the commercial ELISAs out performed western blotting; BioK302: 96% (95% CI: 87% -99%); BioK260:100% (95% CI: 93% -100%) and western blotting: 88% (95% CI: 56% -98%) respectively. In terms of both Se and Sp combined, none of the methods assessed here performed well. As such, when interpreting the results obtained from these ELISAs, the potential impact imperfect Se and / or Sp can have needs to be considered.Despite the poor performance of the BioK302 and BioK260 ELISAs particularly in terms of Se, these tests were used to determine the sero-prevalence of M. bovis antibody in Australian feeder cattle at feedlot induction and draft (approximately 42 days on feed). Paired bovine sera from 1,354 randomly selected animals were sourced from a bovine serum bank (n = 35,160) collected as part of another study, The National Bovine Respiratory Disease Initiative. As the classification of the result of a single serum sample could differ between the BioK302 and BioK260, sero-prevalence estimates were obtained using the individual ELISA results in addition to series and parallel interpretation of the two ELISA kit results. The apparent sero-prevalence of M. bovis-specific antibody at feedlot induction estimates ranged from 2.4% -6.8% (95% CI: 1.3% -8.6%) and the apparent sero-prevalence estimates at draft ranged from 22.4% -44.2% (95% CI: 19.1% -48.6%). The level of sero-conversion to M. bovis between induction and draft was also determined. Only animals that were classified as sero-negative at induction were included in sero-conversion analyses. In the current study, 24.3% (95% iii CI: 20.8% -27.7%) and 19.5% (95% CI: 17.3% -21.7%) of animals sero-converted to M. bovis between induction and draft based on the BioK302 and dichotomised BioK260 ELISA results respectively.Using these ELISA results, an assessment of putative risk factors associated with sero-positivity at feedlot induction or sero-conversion between induction and draft was performed. An increased risk of sero-positivity at feedlot induction was found to be associated with; (i) history of exposure to a saleyard at least 27 d...

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

Zadoks

Middleton

McDougall³

et al. 2011

J Mammary Gland Biol Neoplasia

359

291

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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

Use of a novel real-time PCR technique to monitor and quantitate Mycoplasma bovis infection in cattle herds with mastitis and respiratory disease

Cited by 77 publications

References 17 publications

Bovine Respiratory Syndrome (BRD) Etiopathogenesis, Diagnosis and Control

Bovine Respiratory Syndrome (BRD) Etiopathogenesis, Diagnosis and Control

Mycoplasma bovis in Australian feeder cattle

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

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