Abstract.A real-time polymerase chain reaction (PCR) assay using hybridization probes on a LightCycler platform was developed for detection of Mycoplasma bovis from individual bovine mastitis milk and pneumonic lung tissues. The detection limit was 550 colony forming units (cfu)/ml of milk and 650 cfu/25 mg of lung tissue. A panel of bovine Mycoplasma and of other bovine-origin bacteria were tested; only M. bovis strains were positive, with a melting peak of 66.6ЊC. Mycoplasma agalactiae PG2 was also positive and could be distinguished because it had a melting peak of 63.1ЊC. In validation testing of clinical samples, the relative sensitivity and specificity were 100% and 99.3% for individual milks and 96.6% and 100% for the lung tissue. Using M. bovis real-time PCR, the M. bovis culture-positive milk samples were estimated to contain between 5 ϫ 10 4 and 7.7 ϫ 10 8 cfu/ml and the M. bovis culture-positive lungs between 1 ϫ 10 3 and 1 ϫ 10 9 cfu/25 mg. Isolation, confirmed with the real-time PCR and colony fluorescent antibody test, showed that at the herd level, the proportion of samples positive for M. bovis isolation in mastitis milk samples submitted to the Mastitis Laboratory, Animal Health Laboratory, University of Guelph, Ontario, Canada, was 2.4% (5/201). We conclude that this probe-based real-time PCR assay is a sensitive, specific, and rapid method to identify M. bovis infection in bovine milk and pneumonic lungs.

The findings implied spread of M bovis among calves and suggested that host factors and copathogens may determine disease outcomes in infected calves. Chronic pulmonary infection with M bovis may represent a dynamic situation of bacterial clearance and reinfection with strains of different AFLP type, rather than continuous infection with a single clone. These findings impact our understanding of why cattle with chronic pneumonia and polyarthritis syndrome inadequately respond to antimicrobial treatment.

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The efficacy of moxidectin 2 per cent equine gel against naturally acquired strongyle infections was assessed in 18 ponies which had grazed on contaminated pasture before being housed for eight weeks. Twenty-four hours before the treatment, two randomly selected ponies were euthanased and their worm burdens were determined. Eight of the remaining 16 ponies were treated with moxidectin 2 per cent gel while the other eight were given a placebo gel. Eight weeks later the 16 animals were necropsied and their worm burdens established. A 100 per cent efficacy was recorded against adult and lumenal L4 cyathostomes and adult Strongylus and Triodontophorus species. Digest recoveries of larval cyathostomes indicated a 90.8 per cent (P<0.002) reduction in early L3 and a 99.9 per cent (P<0.001) reduction in developing stages. There was a reduction in faecal egg output of between 96 and 100 per cent in the treated animals compared with the controls.

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