Andrew Pointon

A baseline survey of the microbiological quality of chicken portions and carcasses at retail in two Australian states (2005 to 2006).

Raw poultry products were purchased from the retail market place in two Australian states, New South Wales (n = 549) and South Australia (n = 310). The products sampled on a proportional volume basis were chicken portions with the skin off or skin on, in bulk or tray packs, and whole carcasses. They were collected from butcher shops, supermarkets, and specialty stores from urban areas during the winter (2005) and summer (2006) months. The samples were analyzed to determine the prevalence and concentration of Escherichia coli, Salmonella, and Campylobacter spp. in addition to total viable counts. Salmonella was found in 47.7 and 35.5% of retail chicken samples (35.3 and 21.9% were the less virulent Salmonella Sofia), at mean counts of -1.42 and -1.6 log MPN/cm2 in New South Wales and South Australia, respectively. Campylobacter was found in 87.8 and 93.2% of samples at mean counts of 0.87 and 0.78 log CFU/cm2, respectively. In both states in both seasons, the mean total viable count was 5 log CFU/cm2. On whole birds, E. coli was detected in all winter samples and on 92.9 and 85.7% of summer samples in New South Wales and South Australia, respectively; the log of the geometric mean per square centimeter was 0.5 in winter and slightly lower in summer. On chicken portions, E. coli was detected in around 90% of winter samples in both states, and in summer on 75.1 and 59.6% of samples in New South Wales and South Australia, respectively. The log of the geometric mean CFU per square centimeter for E. coli was 0.75 and 0.91 in winter, and 0.66 and 0.5 in summer in New South Wales and South Australia, respectively.

A Qualitative Assessment of Toxoplasma gondii Risk in Ready-to-Eat Smallgoods Processing

Toxoplasma gondii is one of the most common parasitic infections of humans and other warm-blooded animals. In most adults, it does not cause serious illness, but severe disease may result from infection in fetuses and immunocompromised people. Consumption of raw or undercooked meats has consistently been identified as an important source of exposure to T. gondii. Several studies indicate the potential failure to inactivate T. gondii in the processes of cured meat products, This article presents a qualitative risk-based assessment of the processing of ready-to-eat smallgoods, which include cooked or uncooked fermented meat, pâté, dried meat, slow cured meat, luncheon meat, and cooked muscle meat including ham and roast beef. The raw meat ingredients are rated with respect to their likelihood of containing T. gondii cysts and an adjustment is made based on whether all the meat from a particular source is frozen. Next, the effectiveness of common processing steps to inactivate T. gondii cysts is assessed, including addition of spices, nitrates, nitrites and salt, use of fermentation, smoking and heat treatment, and the time and temperature during maturation. It is concluded that processing steps that may be effective in the inactivation of T. gondii cysts include freezing, heat treatment, and cooking, and the interaction between salt concentration, maturation time, and temperature. The assessment is illustrated using a Microsoft Excel-based software tool that was developed to facilitate the easy assessment of four hypothetical smallgoods products.

Viability of Listeria monocytogenes in co-culture with Acanthamoeba spp.

Listeria monocytogenes is a human pathogen, ubiquitous in the environment, and can grow and survive under a wide range of environmental conditions. It contaminates foods via raw materials or food-processing environments. However, the current knowledge of its ecology and, in particular, the mode of environmental survival and transmission of this intracellular pathogen remains limited. Research has shown that several intracellular pathogens are able to survive or replicate within free-living amoebae. To examine the viability of L. monocytogenes in interaction with Acanthamoeba spp., bacteria were co-cultured with three freshly isolated amoebae, namely Acanthamoeba polyphaga, Acanthamoeba castellanii and Acanthamoeba lenticulata. The survival of bacteria and amoebae was determined using culture techniques and microscopy. Under the experimental conditions used, all amoebae were able to eliminate bacteria irrespective of the hly gene. Bacteria did not survive or replicate within amoeba cells. However, extra-amoebic bacteria grew saprophytically on materials released from amoebae, which may play an important role in the survival of bacteria under extreme environmental conditions.

Using slaughter inspections to evaluate sarcoptic mange infestation of finishing swine.

Sarcoptic mange is one of the common swine diseases worldwide. Although mange-free populations can be established with caesarean derived stock, by herd repopulation programmes or by eliminating mange with ivermectin, mange remains prevalent in many countries. Field and experimental studies indicate that hypersensitive mange is detrimental to performance of growing pigs. Typically, producers tolerate mange infestation in their herds and control measures are often haphazard. This tolerance to mange infestation is attributable to the covert nature of the losses (reduced growth rate and feed efficiency without mortality) and to the fact that clinical signs of hypersensitive mange (pruritus) are usually viewed as normal. Lack of tools to evaluate mange severity in pigs and to demonstrate its importance has hindered the efforts of veterinarians to control the disease. Traditionally, veterinarians have used slaughter inspections to assess respiratory diseases such as enzootic pneumonia and atrophic rhinitis. Much of the value of slaughter inspections is as a tool with which veterinarians can educate and motivate their clients to improve disease control measures. The potential for evaluating hypersensitive mange by inspecting slaughtered pigs for lesions of papular dermatitis was recognised some time ago, but quantitative evaluation of the reliability of this approach has been lacking. We have conducted several studies in Australia, the USA, Canada, Europe and Latin America to evaluate associations between Sarcoptes infestation and the severity of papular dermatitis at slaughter, using a simple ordinal scale for classifying carcasses. Our initial field and experimental data in Australia indicated the specificity of localised dermatitis to be in the order of 75-80%, but that the generalised dermatitis was highly specific (> 98%) for mange. Subsequent studies in the US Midwest yielded almost identical results, and indicated that the method may also have some utility for surveillance of mange-free herds. Results from other locations invariably have shown significant associations between dermatitis lesions and mange infestation. Relative to other methods such as skin scrapings and monitoring pruritus, this method is simple and relatively objective, and should be considered for routine inclusion in slaughter inspection protocols.

Mechanism involved in phagocytosis and killing of Listeria monocytogenes by Acanthamoeba polyphaga.

Intra-cellular pathogen, Listeria monocytogenes, is capable of invasion and survival within mammalian cells. However, Acanthamoeba polyphaga trophozoites phagocytose and rapidly degrade Listeria cells. In order to provide more information on amoeba phagocytosis and killing mechanisms, this study used several inhibitor agents known to affect the phagocytosis and killing of bacteria by eukaryotes. Amoebae were pre-treated with mannose, cytochalasin D, wortmannin, suramin, ammonium chloride, bafilomycin A and monensin followed by co-culture with bacteria. Phagocytosis and killing of bacterial cells by amoeba trophozoites was assessed using plate counting methods and microscopy. The data presented indicates that actin polymerisation and cytoskeletal rearrangement are involved in phagocytosis of L. monocytogenes cells by A. polyphaga trophozoites. Further, both phagosomal acidification and phagosome–lysosome fusion are involved in killing and degradation of L. monocytogenes cells by A. polyphaga. However, the mannose-binding protein receptor does not play an important role in uptake of bacteria by amoeba trophozoites. In conclusion, this data reveals the similar principles of molecular mechanisms used by different types of eukaryotes in uptake and killing of bacteria.

Listeria monocytogenes does not survive ingestion by Acanthamoeba polyphaga.

Listeria monocytogenes is a ubiquitous bacterium capable of infecting humans, particularly pregnant women and immunocompromised individuals. Although the intracellular invasion and pathogenesis of listeriosis in mammalian tissues has been well studied, little is known about the ecology of L. monocytogenes , and in particular the environmental reservoir for this bacterium has not been identified. This study used short-term co-culture at 15, 22 and 37 degrees C to examine the interaction of L. monocytogenes strains with Acanthamoeba polyphaga ACO12. Survival of L. monocytogenes cells phagocytosed by monolayers of trophozoites was assessed by culture techniques and microscopy. A. polyphaga trophozoites eliminated bacterial cells within a few hours post-phagocytosis, irrespective of the incubation temperature used. Wild-type L. monocytogenes and a phenotypic listeriolysin O mutant were unable to either multiply or survive within trophozoites. By contrast, Salmonella enterica serovar Typhimurium C5 cells used as controls were able to survive and multiply within A. polyphaga trophozoites. The data presented indicate that A. polyphaga ACO12 is unlikely to harbour L. monocytogenes, or act as an environmental reservoir for this bacterium.

Risk-based evaluation of postmortem inspection procedures for pigs in Australia

The results of traditional (incision) and risk-based (visual) postmortem inspection procedures were compared on groups of approximately 30,000 pigs. The performance characteristics used as a basis for comparison included the non-detection rates of grossly detectable abnormalities, the microbiological contamination rates of carcases and boned product, the association of reactive lymph nodes with carcase condemnation and the achievement of ‘finished product standards’ for ‘wholesomeness’ It was estimated that 6 per cent of all cases of abscessation and 28 per cent of all cases of arthritis were undetected by the traditional method, and the comparable figures for the risk-based procedure were 19 per cent and 39 per cent. However, when the rates of contamination of undetected abnormalities with foodborne hazards and other carcase contamination parameters were taken into account, it was concluded that both inspection systems were likely to result in a very similar level of consumer protection. Any increase in potential exposure to foodborne hazards in the abnormalities undetected by risk-based inspection would be insignificant in comparison with the potential exposure to foodborne hazards resulting from contaminated ‘normal’ lymph nodes and carcase surfaces. There were no statistically significant differences between the two procedures in the contamination rates of pre-chill carcases or boned retail products with Salmonella and Yersinia species.

Slaughterfloor Decontamination of Pork Carcases with Hot Water or Acidified Sodium Chlorite – A Comparison in Two Australian Abattoirs

A decontamination trial on the effectiveness of hot water or acidified sodium chlorite (SANOVA™) treatment on Salmonella spp., Escherichia coli and Total Viable Count (TVC) was undertaken on pork carcases prior to primary chilling in two large pork abattoirs in Australia using belly‐strip excision sampling. A total of 123 samples from Abattoir A and 400 samples from Abattoir B were cultured and analysed. Test pigs were selected from herds with a known high level of on‐farm Salmonella infection. At Abattoir A, Salmonella spp. were not isolated from carcases. The prevalence of E. coli on control carcases was 92.9% compared with 9.8% for hot water and 12.5% for SANOVA™ treated carcases. The mean log10E. coli concentration for control carcases was 0.89 cfu/gram, compared with −0.83 cfu/gram from hot water and −0.75 cfu/gram from SANOVA™ treated carcases. The mean log10 TVC for control carcases was 4.06 compared with 1.81 cfu/gram for hot water and 2.76 cfu/gram for SANOVA™ treated carcases. At Abattoir B, the prevalence of Salmonella on control carcases was 16% compared with 2.7% for hot water and 7.0% for SANOVA™ treated carcases. The prevalence of E. coli on control carcases was 69.3% compared with 22% for hot water and 30% for SANOVA™ treated carcases. The mean log10E. coli concentration for control carcases was 0.45 cfu/gram, compared with −0.65 cfu/gram from hot water and −0.60 cfu/gram from SANOVA™ treated carcases. The mean log10 TVC for control carcases was 3.00 cfu/gram compared with 2.10 cfu/gram for hot water and 2.53 cfu/gram for SANOVA™ treated carcases. The reductions in prevalence and mean log10 concentrations in the present trial were all found to be statistically significant and indicate that carcases decontamination with either hot water or SANOVA™ are effective risk management options immediately available to the pork industry.

Microbial profiles of carcasses and minced meat from kangaroos processed in South Australia

The microbiological profiles of kangaroo carcasses and minced meat at game meat processing plants in South Australia were determined in surveys undertaken in 2002 and 2004. In 2002 mean values for log(10) total viable counts (TVC) on carcasses at individual plants ranged from 0.9 to 3.9 log(10) cfu/cm(2), with the mean for all plants being 2.3 log(10) cfu/cm(2). In 2004 the between plant range was narrower, by about 1 log unit, and the mean value for carcasses at all plants was 1.2 log(10) cfu/cm(2). Minced kangaroo meat, was sampled in 2002 only. The overall mean log(10) TVC was 3.9 log(10) cfu/g, with mean counts at individual plants ranging from 3.1 to 4.6 log(10) cfu/g. The overall prevalence of E. coli was 70%, with mean numbers of 2.1 log(10) cfu/g on positive samples. Salmonella was not detected in any of 60 samples from carcasses in 2002. However, in 2004 Salmonella was detected in 4/385 samples (1.04%, 95% CI: 0.28%-2.64%). In minced kangaroo meat, Salmonella was detected in 9/50 (18%, 95% CI: 9%-31%) samples. The abdominal cavity, sampled in 2004, was found to be highly contaminated, with E. coli isolated from 46% of samples and the mean number for positive samples being 2.7 log(10) cfu/cm(2); Salmonella was isolated from 14/120 (12%; 95% CI: 6.52%-18.80%) of abdominal cavities. The practice of collecting carcasses together and pushing grouped carcasses into the chiller likely leads to cross contamination of carcasses from the abdominal cavities of others. To align results of sampling by swabbing for domestic purposes with excision sampling, required for export purposes, both methods were used to sample opposite sides of each of the 50 carcasses sampled in 2004. The results obtained with the two methods of sampling were similar.

Longitudinal study of Salmonella 1,4,[5],12:i:- shedding in five Australian pig herds

The shedding patterns of Salmonella spp. and MLVA profiles of Salmonella enterica subspecies enterica (I) serotype 1,4,[5],12:i:- were monitored in a 12-month longitudinal observational study of five pig herds to inform management; provide indications of potential hazard load at slaughter; and assist evaluation of MLVA for use by animal and public health practitioners. Twenty pooled faecal samples, stratified by age group, were collected quarterly. When Salmonella was cultured, multiple colonies were characterized by serotyping and where S. Typhimurium-like serovars were confirmed, isolates were further characterized by phage typing and multiple locus variable number tandem repeat analysis (MLVA). Salmonella was detected in 43% of samples. Salmonella 1,4,[5],12:i- was one of several serovars that persisted within the herds and was found among colonies from each production stage. Virtually all Salmonella 1,4,[5],12:i:- isolates were phage type 193, but exhibited 12 different, closely-related MLVA profiles. Salmonella 1,4,[5],12:i:- diversity within herds was low and MLVA profiles were stable indicating colonization throughout the herds and suggesting each farm had an endemic strain. High prevalence of S. 1,4,[5],12:i:- specific shedding among terminal animals indicated high hazard load at slaughter, suggesting that primary production may be an important pathway of S. 1,4,[5],12:i:- into the human food chain, this has implications for on-farm management and the application and targeting control measures and further evidence of the need for effective process control procedures to be in place during slaughter and in pork boning rooms. These findings have implications for animal health and food safety risk mitigation and risk management.