J.A. Hudson

Bacteriophages as biocontrol agents in food

Bacteriophages possess attributes that appear to be attractive to those searching for novel ways to control foodborne pathogens and spoilage organisms. These phages have a history of safe use, can be highly host specific, and replicate in the presence of a host. Campylobacter, Salmonella, and Listeria monocytogenes and various spoilage organisms have responded to phage control on some foods. However, the use of phages as biocontrol agents is complicated by factors such as an apparent requirement for a threshold level of host before replication can proceed and by suboptimal performance, at best, at temperatures beneath the optimum for the host. This review is a summary of the information on these issues and includes brief descriptions of alternative phage-based strategies for control of foodborne pathogens.

Enumeration of Campylobacter in New Zealand recreational and drinking waters

Aims: To use a published polymerase chain reaction (PCR) method for the detection and identification of thermotolerant Campylobacter species (Camp. jejuni, Camp. coli and Camp. lari) in tandem with a Most Probable Number (MPN) technique to enumerate these species in water samples.

Phage inactivation of foodborne pathogens on cooked and raw meat

Phages infecting Salmonella Typhimurium PT160 and Campylobacter jejuni were added at a low or high (10 or 10(4)) multiplicity of infection (MOI) to either low or high (<100 or 10(4)cm(-2)) densities of host bacteria inoculated onto raw and cooked beef, and incubated at 5 and 24 degrees C to simulate refrigerated and room temperature storage. Counts of host bacteria were made throughout the incubation period, with phages being counted at the first and last sampling times. Host inactivation was variable and depended on the incubation conditions and food type. Significant host inactivations of the order of 2-3 log(10)cm(-2) at 5 degrees C and >5.9 log(10)cm(-2) at 24 degrees C were achieved compared to phage-free controls using the Salmonella phage under optimal conditions (high host cell density and MOI). These results alongside those already published indicate that phages may be useful in the control for foodborne pathogens.

The occurrence of Campylobacter subtypes in environmental reservoirs and potential transmission routes

Aim:  To identify potential reservoirs and transmission routes of human pathogenic Campylobacter spp.

Seasonal variation of Campylobacter types from human cases, veterinary cases, raw chicken, milk and water

During August 1996 (winter) and February 1997 (summer), a total of 180 Campylobacter isolates from a restricted geographical area were obtained from human and veterinary cases, raw milk and chicken, and untreated water. Isolates were typed by Penner serotyping and pulsed‐field gel electrophoresis (PFGE) of restriction enzyme‐produced DNA fragments. Differences were noted between the August and February serotypes, with the most, and fourth most frequently isolated serotypes in February being completely absent in August. Two other serotypes were more frequently found in the February isolates, while the reverse was true for two others. In contrast to the serotyping data, one PFGE restriction profile type was dominant in both seasons, and the pattern of distribution of isolates among the other restriction patterns was similar. Five groups of isolates in each month were indistinguishable by both typing methods. Only one group was common to both months. Another group, which was absent in August, dominated the February isolates. Marked differences in the types isolated in the two seasons were therefore evident. Some isolates from human cases were indistinguishable from others isolated from water and raw chicken, indicating possible routes of infection for humans.

Series of incidents of Listeria monocytogenes non‐invasive febrile gastroenteritis involving ready‐to‐eat meats

Aims: A series of cases and outbreaks of febrile noninvasive gastrointestinal disease involving 31 identified cases was investigated in terms of the numbers and types of Listeria monocytogenes present in the suspect foods (ready‐to‐eat meats) and clinical samples from cases.

Prevalence, Numbers, and Subtypes of Campylobacter jejuni and Campylobacter coli in Uncooked Retail Meat Samples

A national quantitative survey of Campylobacter jejuni and Campylobacter coli in 1,011 uncooked retail meat samples (beef, unweaned veal, chicken, lamb and mutton, and pork) was undertaken from August 2003 to June 2004 to establish baseline proportionality data. The presence, number, and type of Campylobacter present in each sample was assessed. Prevalences of C. jejuni and C. coli were 89.1% in chicken, 9.1% in pork, 6.9% in lamb and mutton, 3.5% in beef, and 10% in unweaned veal. C. jejuni was identified in the majority of positive samples (246 of 259). In chicken samples positive for C. jejuni, 40.2% had counts of <0.3 most probable number (MPN)/g, 50.5% had 0.3 to 10.0 MPN/g, 8.8% had 10.1 to 50.0 MPN/g, and 0.5% had 110 MPN/g. In other meats (49 samples), Campylobacter counts were < or = 0.3 MPN/g, except for one unweaned veal sample at > 10.9 MPN/g. Penner serotyping and SmaI macrorestriction genotyping using pulsed-field gel electrophoresis with 247 isolates revealed 17 Penner serotypes and 56 electrophoresis profiles. Seven Penner serotypes (HS1 complex, 2, 4 complex, 6, 11, 27, and 42) were represented by 10 or more isolates from chicken. When data from both typing methods were combined, 62 sero-genotypes were generated. In a comparison of these sero-genotypes with historical data for isolates from human cases, 71% of the beef isolates, 50% of the lamb and mutton isolates, 50% of the pork isolates, 41% of the chicken isolates, and 25% of the unweaned veal isolates were common to both sources. These results provide baseline proportionality profiles of Campylobacter in these five meats and will facilitate exposure assessment in combination with other information such as consumption data and subsequent quantitative risk assessment.

Rapid detection of Listeria monocytogenes in ham samples using immunomagnetic separation followed by polymerase chain reaction

Aims: To develop a 24‐h system for the detection of Listeria monocytogenes in ham.

Conversion of oleic acid to 10-hydroxystearic acid by two species of ruminal bacteria.

Bacteria able to convert oleic acid to 10-hydroxystearic acid were isolated from the ovine rumen. The solid hydroxy fatty acid produced from bacterial fermentations containing oleic acid was recovered by filtration, extraction into ether and crystallisation. The identity of the product was confirmed by HPLC and gas chromatography/mass spectrometry. One 10-hydroxystearic-acid-producing bacterial group was represented by two strains of an anaerobic gram-negative curved rod with tufts of flagella on the concave surface of the cell. The morphology and other characteristics enabled the strains to be tentatively identified as Selenomonas ruminantium. Another bacterium capable of the same transformation, represented by two strains of a facultatively anaerobic gram positive chain-forming coccus, was identified as Enterococcus faecalis. Since unsaturated fatty acids entering the rumen are normally hydrogenated, hydration of oleic acid represents an alternative fate of unknown significance in vivo.

Thermus filiformis sp. nov., a filamentous caldoactive bacterium

In a preliminary investigation the isolation of a caldoactive filamentous microorganism from a New Zealand hot spring was reported. This organism is described here as a new species belonging to the genus Thermus, namely, Thermus filiformis, based on ultrastructural, phenotypic, and anomalous Gram type characteristics. The cell wall of T. filiformis resembles that of Thermus aquaticus apart from the presence of an extra layer. The Thermus species tested, including T. filiformis, are negative for the aminopeptidase test, which is unusual for a gram-negative genus. T. filiformis is nonproteolytic, unlike most other Thermus strains, and also differs radically from other strains in morphology when it is observed by using phase-contrast microscopy. The single strain of the species has been deposited with the American Type Culture Collection as strain ATCC 43280T (T = type strain).