The use of PCR for the identification and characterisation of bacteriocin genes from bacterial strains isolated from rumen or caecal contents of cattle and sheep.
PCR primers were designed to amplify the gene that encodes bovicin 255 from Streptococcus gallolyticus LRC0255 and the bacteriocin genes from Butyrivibrio fibrisolvens strains AR10 and OR79A (bviD and bvi79A) in order to screen for their incidence in rumen and caecal B. fibrisolvens and Streptococcus bovis-like isolates from New Zealand and North American ruminants. None of the B. fibrisolvens-like strains (n=34) isolated from New Zealand or North America had the genes encoding for butyrivibriocins AR10 (bviD) or OR79 (bvi79A). However, seven S. bovis isolates from New Zealand ruminants and three from North American animals had the bovicin 255 gene. Sequence comparison of cloned bovicin 255 PCR products indicated a 92.9-95.7% similarity to that of the corresponding bovicin 255 gene sequence of S. gallolyticus. Four of the New Zealand bovicin 255 positive S. bovis isolates were from the caecal contents of the same sheep and had identical PFGE profiles. Two other S. bovis isolates sharing the same PFGE profile were isolated from a separate animal from the same flock. PFGE analysis of the North American strains indicated that all three were closely related as two of three had identical PFGE profiles with the remaining isolate differing only by a single band position. The 16S rRNA gene sequences of the 10 isolates were at least 99.8% identical to S. bovis. All 10 S. bovis isolates having the gene for bovicin 255 produced bacteriocin activity that inhibited the growth of Peptostreptococcus anaerobius D1 in a deferred antagonism plating (DAP) assay. Certain S. bovis isolates obtained from ruminants have bacteriocin activity associated with a distinct bovicin 255 gene sequence but it appears that bacteriocin production by the rumen anaerobe B. fibrisolvens may be uncommon in strains isolated from cattle and sheep in New Zealand.
Cookson
Adrian L
AL
Rumen Microbial Functional Genomics, Nutrition and Behaviour Group, AgResearch, Grasslands Research Centre, Tennent Drive, Palmerston North, New Zealand. adrian.cookson@agresearch.co.nz
Noel
Samantha J
SJ
Kelly
William J
WJ
Attwood
Graeme T
GT
D000595:Amino Acid Sequence; D000818:Animals; D015373:Bacterial Typing Techniques / Q000379:methods; D001430:Bacteriocins / Q000235:genetics*; D045856:Butyrivibrio / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D002417:Cattle; D002432:Cecum / Q000382:microbiology*; D016000:Cluster Analysis; D016172:DNA Fingerprinting / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D016521:Electrophoresis, Gel, Pulsed-Field; D005798:Genes, Bacterial; D005838:Genotype; D008969:Molecular Sequence Data; D009520:New Zealand; D009656:North America; D016133:Polymerase Chain Reaction / Q000379:methods*; D012417:Rumen / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D012756:Sheep; D013291:Streptococcus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification
D001430:Bacteriocins; D017931:DNA Primers; D004269:DNA, Bacterial