Contamination of milk by enterococci and coliforms from bovine faeces

Aim: To determine the contribution of enterococci and coliforms from bovine faeces and teats to contamination of raw milk. Methods: Putative enterococci ( n = 301) and coliforms ( n = 365) were isolated from bovine faeces ( n = 20), cows' teats ( n = 20), the raw milk ( n = 1) and the milking environment ( n = 4) on one farm. The clonal relationships of each bacterial group were investigated using Pulsed- Field Gel Electrophoresis of genomic macrorestriction fragments. Representatives of the different clusters of enterococci were identified by molecular techniques including rep- PCR, SDS protein profiling, Fluorescent Amplified Fragment Length Polymorphism ( FAFLP), phenylalanyl- tRNA synthase ( pheS) sequence analysis and / or 16S rDNA gene sequencing. Coliforms were identified by API 20E strips. Results: The majority of the bovine faecal enterococcal isolates were identified as a potential new species of Aerococcus ( 100 isolates); E. faecium ( 28 isolates), and Aerococcus viridans ( 28 isolates) were also found. All coliform isolates from the bovine faeces were identified as Escherichia coli. The coliforms present in the milk were Hafnia alvei, Serratia liquefaciens, Yersinia enterocolitica and Enterobacter amnigenus. No E. coli, Enterococcus or Aerococcus from the bovine faeces were found in the milk. A single clone of H. alvei was found in the water, the milking equipment and the milk, suggesting that the water was the source of the organism in the milk. No vancomycin- resistant aerococci or enterococci were found while most of the isolates tested showed the presence of at least one virulence gene. The milk- sock retained strains that adhered to particulate faecal material. Coliforms were present at approx. 2 orders of magnitude greater than enterococci in the bovine faeces. Conclusions: The results imply that bovine faeces are not an important source of contamination of raw milk with enterococci or coliforms. Significance and Impact of the Study: The results confirm those of two previous studies ( Gelsomino et al. 2001, Int J Food Microbiol 71, 177 - 188 and Kagkli et al. 2007, Int J Food Microbiol 114, 243 - 251) on two other farms. The three studies show that contamination of milk by enterococci, lactobacilli and coliforms of bovine faecal origin is extremely low. The results also suggest that where raw milk is implicated in food infection, other factors in addition to faecal contamination must be involved.