A multiplex polymerase chain reaction to detect and differentiate Campylobacter fetus subspecies fetus and Campylobacter fetus -species venerealis: use on UK isolates of C. fetus and other Campylobacter spp. Aims: Subspeciation of Campylobacter fetus subsp. fetus (CFF) and Campylobacter fetus subsp. venerealis (CFV) is important for international animal import regulations. Phenotyping can be unreliable, and genotyping by techniques like pulsed field gel electrophoresis is difficult in routine diagnostic laboratories. A PCR subspeciation technique has been reported [Aust Vet J (1997) 75, 827]; we aimed to develop this PCR and investigate its use on UK C. fetus isolates.
Methods and Results:We augmented the PCR with further primers, and tested 76 isolates of C. fetus and 16 isolates of other Campylobacter spp. PCR failed to correlate well with phenotyping, especially for CFV. We characterized the amplicon of the CFV-specific primers (reported as plasmid derived, but unavailable on the public databases); and predicted a parA gene sequence, anticipated to be plasmid-associated. However, although plasmid isolations from selected CFV isolates demonstrated the presence of several plasmids, there was no correlation between plasmid profile and PCR result. Further, the parA sequence was not detected by PCR in any of the plasmid bands. Conclusions: This PCR is not suitable for subspeciation of C. fetus in the UK. The results suggest that this is a reflection of the presence of an unusual clone of CFV currently present in cattle in this country. Significance and Impact of the Study: PCR cannot substitute for phenotyping of C. fetus isolates in the UK. The reasons for failure of PCR genotyping may reflect local strains and/or plasmid profiles. Further study is required to better elucidate molecular sub-speciation of C. fetus.
A natural infection with border disease virus occurred in a flock on low ground in Argyll in the spring of 1984. The outbreak was unusual in that the typical clinical signs of border disease, ie, tremor and, or, fleece changes were not present; manifestations of disease were restricted to abortion and the birth of small weak lambs. The disease was shown to have been introduced to the flock by four healthy ewes persistently infected with border disease virus among a group of 39 purchased in October 1983. Further investigations in late August 1984 detected viraemia in six of seven ill-thriven lambs and four of 24 apparently healthy lambs. Attempted 'natural vaccination' of susceptible sheep by mixing them at grass for three months with groups of ewes and lambs known to contain virus excretors was largely unsuccessful as only four of 22 'sentinel' sheep seroconverted. In October 1984 the persistently infected purchased animals and all that year's lamb crop were removed from the farm. No disease occurred in 1985 when the lambing percentage was 129 per cent compared with 100 per cent in 1984. Two of the four persistently infected purchased ewes were mated at Moredun Research Institute in December 1984 and both produced healthy but persistently infected lambs.
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