Spoilage potential of bacterial species from chilled vacuum-packed lamb.

The objective of this study was to establish whether specific organisms play important roles in the spoilage rate of vacuum-packed (VP) lamb at low storage temperatures. The spoilage potential of representative organisms (n = 13) of the spoilage community of VP lamb were investigated through a series of shelf-life challenge trials. Each isolate was individually inoculated onto sterile (irradiated) and non-sterile (i.e., containing natural microbial community) VP lamb meat. Meat quality was assessed over time by measuring sensorial qualities, bacterial growth and pH. Among all test organisms, Clostridium spp. had the highest spoilage potential and had a major effect on the spoilage rate of VP lamb (based on sensory assessment). C. estertheticum caused premature 'blown pack' spoilage; however, the spoilage was delayed in a community setting. C. putrefaciens and C. algidicarnis caused premature spoilage of VP lamb independently and in a community setting. In contrast, all facultative anaerobes and Pseudomonas sp. tested were not capable of spoiling meat independently or within a community, expect for Carnobacterium divergens and Serratia spp., which spoiled meat prematurely when present in a community. Overall, these results highlight that Clostridium could be one of the main taxa driving the faster rate of quality loss of chilled VP lamb compared to beef. This research can help to inform opportunities for shelf-life extension by targeting organisms with 'high' spoilage potential, such as Clostridium.

Effect of abattoir, livestock species and storage temperature on bacterial community dynamics and sensory properties of vacuum packaged red meat.

Shelf life of red meat is influenced by a number of intrinsic and extrinsic factors making its prediction challenging. Here we investigated the influence of geographically distant abattoir facilities and storage temperature relevant to commercial supply chain on the shelf lives of vacuum packaged (VP) beef and lamb meat. Samples of VP beef and lamb were analysed for surface pH, total viable counts, lactic acid bacterial counts, sensory properties, and associated bacterial community using Illumina MiSeq based 16S rRNA gene amplicon sequencing over a period of >200 days. The consistent 0.41 pH unit difference between beef and lamb was found to have a profound effect on bacterial community diversity and composition, bacterial growth rates and the rate of loss of sensory quality. Though different community structures were derived from different abattoir source, bacterial growth rate and rate of sensory quality deterioration were found to be comparable for individual meat type. The greatest variation in rates was found resulting from storage temperature and livestock species themselves. Our findings indicate that bacterial growth and sensory quality loss are essentially predictable when considering their temperature dependency, however for successful meat export validation of shelf life predictive models is required due to stochastic variation in abattoir seeded bacterial populations.

Consumer Safety Considerations of Skin and Oral Microbiome Perturbation.

Microbiomes associated with human skin and the oral cavity are uniquely exposed to personal care regimes. Changes in the composition and activities of the microbial communities in these environments can be utilized to promote consumer health benefits, for example, by reducing the numbers, composition, or activities of microbes implicated in conditions such as acne, axillary odor, dandruff, and oral diseases. It is, however, important to ensure that innovative approaches for microbiome manipulation do not unsafely disrupt the microbiome or compromise health, and where major changes in the composition or activities of the microbiome may occur, these require evaluation to ensure that critical biological functions are unaffected. This article is based on a 2-day workshop held at SEAC Unilever, Sharnbrook, United Kingdom, involving 31 specialists in microbial risk assessment, skin and oral microbiome research, microbial ecology, bioinformatics, mathematical modeling, and immunology. The first day focused on understanding the potential implications of skin and oral microbiome perturbation, while approaches to characterize those perturbations were discussed during the second day. This article discusses the factors that the panel recommends be considered for personal care products that target the microbiomes of the skin and the oral cavity.

Application of chlorine dioxide and peroxyacetic acid during spray chilling as a potential antimicrobial intervention for beef carcasses.

Enteric pathogens such as Shiga-toxin producing Escherichia coli (STEC) and Salmonella spp. continue to be a major food safety concern for the beef industry. Currently, no single method is completely effective in controlling these pathogens during carcass processing. Previous research, however, suggested that STEC might become more susceptible to oxidative damage when exposed to carcass chilling (King et al., 2016). We aimed to test that hypothesis by evaluating the antimicrobial effects of an oxidant (chlorine dioxide, ClO2 or peroxyacetic acid, PAA) on beef meat during a simulated spray chilling process (sprayed for 4 s every 15 min for 36 cycles) and/or when applied (sprayed for 144 s) prior to spray chilling with water. In all experiments, the inactivating effects of oxidants were greatest on fat surfaces and much less effective on lean surfaces. ClO2 at 15 ppm, a non-lethal level for E. coli under optimal growth conditions, caused higher log reductions in E. coli numbers (approximately 3-log reduction) when applied during spray chilling than when applied immediately prior to 'normal' spray chilling (approximately 1-log reduction). This confirms the hypothesis that E. coli are more susceptible to oxidative stress during spray chilling. In subsequent studies, both ClO2 and PAA at lethal levels (at ≥20 and ≥ 200 ppm, respectively) applied during spray chilling resulted in pronounced inactivation of both E. coli and Salmonella enterica strains, achieving a ≥4-log reduction at the end of chilling. These results indicate that an oxidant-based application during spray chilling as an antimicrobial intervention could be effective to minimise the problems associated with enteric pathogen contamination on beef meat.

Innovative processes and technologies for modified atmosphere packaging of fresh and fresh-cut fruits and vegetables.

Modified atmosphere packaging (MAP) technology has been commercially viable since the 1970s. Currently, MAP is extensively used worldwide to preserve the quality and extend the shelf-life of whole fresh fruits and vegetables, but is also increasingly used to extend the shelf-life of minimally processed fresh fruit and vegetables. This review discusses new processes and technologies that can be used to improve quality preservation and consumer acceptability of minimally processed produce where high respiration rates and challenging degradation processes operate. New packaging innovations are enabling producers and retailers to further maintain quality for longer. Innovative approaches to extend shelf-life include active MAP with differentially permeable films, films that incorporate antimicrobial properties, edible coatings that confer barriers properties, and the use of non-traditional gases to modify respiration. Intelligent packaging using integrated sensor technologies that can indicate maturity, ripeness, respiration rate and spoilage are also appearing. This review demonstrates that preservation technologies and associated packaging developments that can be combined with modified atmosphere are constantly evolving technology platforms. Adoption of combinations of technology improvements will be critical in responding to commercial trends towards more minimally processed fresh-cut and ready-to-eat fruit and vegetable products, which require specialized packaging solutions.

Physiological Response of Escherichia coli O157:H7 Sakai to Dynamic Changes in Temperature and Water Activity as Experienced during Carcass Chilling.

Enterohemeorrhagic Escherichia coli is a leading cause of foodborne illness, with the majority of cases linked to foods of bovine origin. Currently, no completely effective method for controlling this pathogen during carcass processing exists. Understanding how this pathogen behaves under those stress conditions experienced on the carcass during chilling in cold air could offer opportunities for development or improvement of effective decontamination processes. Therefore, we studied the growth kinetics and physiological response of exponential phase E. coli O157:H7 Sakai cultures upon an abrupt downshift in temperature and water activity (from 35 °C aw 0.993 to 14 °C aw 0.967). A parallel Biolog study was conducted to follow the phenotypic responses to 190 carbon sources. Exposure of E. coli to combined cold and water activity stresses resulted in a complex pattern of population changes. This pattern could be divided into two main phases, including adaptation and regrowth phases, based on growth kinetics and clustering analyses. The transcriptomic and proteomic studies revealed that E. coli exhibited a "window" of cell susceptibility (i.e. weaknesses) during adaptation phase. This included apparent DNA damage, the downregulation of molecular chaperones and proteins associated with responses to oxidative damage. However, E. coli also displayed a transient induction in the RpoE-controlled envelope stress response and activation of the master stress regulator RpoS and the Rcs phosphorelay system involved in colanic acid biosynthesis. Increased expression was observed for several genes and/or proteins involved in DNA repair, protein and peptide degradation, amino acid biosynthesis, and carbohydrate catabolism and energy generation. Furthermore, the Biolog study revealed reduced carbon source utilization during adaptation phase, indicating the disruption of energy-generating processes. This study provides insight into the physiological response of E. coli during exposure to combined cold and water activity stress, which could be exploited to enhance the microbiological safety of carcasses and related foods.

Quantitative Microbial Risk Assessment of Salmonellosis from the Consumption of Australian Pork: Minced Meat from Retail to Burgers Prepared and Consumed at Home.

Pork burgers could be expected to have an elevated risk of salmonellosis compared to other pork products due to their comminuted nature. A stochastic risk assessment was performed to estimate the risk of salmonellosis from Australian pork burgers and considered risk-affecting factors in the pork supply chain from retail to consumption at home. Conditions modeled included prevalence and concentration of Salmonella in pork mince, time and temperature effects during retail, consumer transport, and domestic storage and the effect of cooking, with the probability of illness from consumption estimated based on these effects. The model was two-dimensional, allowing for the separation of variability and uncertainty. Potential changes to production practices and consumer behaviors were examined through alternative scenarios. Under current conditions in Australia, the mean risk of salmonellosis from consumption of 100 g pork burgers was estimated to be 1.54 × 10 - 8 per serving or one illness per 65,000,000 servings consumed. Under a scenario in which all pork mince consumed is served as pork burgers, and with conservative (i.e., worst-case) assumptions, 0.746 cases of salmonellosis per year from pork burgers in Australia were predicted. Despite the adoption of several conservative assumptions to fill data gaps, it is predicted that pork burgers have a low probability of causing salmonellosis in Australia.

Culture-dependent and culture-independent assessment of spoilage community growth on VP lamb meat from packaging to past end of shelf-life.

Packaging and storage temperature are important factors that influence the shelf-life of vacuum packed (VP) meat. In this study the shelf-life of VP bone-in lamb hind shanks stored at 8 °C and -1.2 °C was determined in parallel to analyses of starting and eventual spoilage bacterial communities via Illumina MiSeq based 16S rRNA amplicon sequencing. The mean total viable counts (TVC) and lactic acid bacterial viable counts (LAB) were observed to increase to log 7.5 CFU/cm2 and 7 CFU/cm2 after 6 and 42 days at 8 °C and -1.2 °C and stayed stable until shelf-life termination after 13 and 124 days, respectively. The sequence data showed initial communities were patchily distributed and were mainly derived from skin microbiome taxa likely prevalent within the abattoir. A broad diversity of VP meat associated specific spoilage organisms (SSO) were comparatively abundant in this initial population. Overtime meat spoilage communities developed a distinctive and stable microbiome. At -1.2 °C SSO included mainly Carnobacterium, Yersinia and Clostridium spp. while at 8 °C SSO expanded to include Hafnia, Lactococcus, Providencia spp. Growth curves inferred from the sequence data after taking into account rRNA copy number suggested that SSO growth rates were consistent with overall growth rates determined from TVC and LAB data and are predictable.

Integrated transcriptomic and proteomic analysis of the physiological response of Escherichia coli O157:H7 Sakai to steady-state conditions of cold and water activity stress.

An integrated transcriptomic and proteomic analysis was undertaken to determine the physiological response of Escherichia coli O157:H7 Sakai to steady-state conditions relevant to low temperature and water activity conditions experienced during meat carcass chilling in cold air. The response of E. coli during exponential growth at 25 °C a(w) 0.985, 14 °C a(w) 0.985, 25 °C a(w) 0.967, and 14 °C a(w) 0.967 was compared with that of a reference culture (35 °C a(w) 0.993). Gene and protein expression profiles of E. coli were more strongly affected by low water activity (a(w) 0.967) than by low temperature (14 °C). Predefined group enrichment analysis revealed that a universal response of E. coli to all test conditions included activation of the master stress response regulator RpoS and the Rcs phosphorelay system involved in the biosynthesis of the exopolysaccharide colanic acid, as well as down-regulation of elements involved in chemotaxis and motility. However, colanic acid-deficient mutants were shown to achieve comparable growth rates to their wild-type parents under all conditions, indicating that colanic acid is not required for growth. In contrast to the transcriptomic data, the proteomic data revealed that several processes involved in protein synthesis were down-regulated in overall expression at 14 °C a(w) 0.985, 25 °C a(w) 0.967, and 14 °C a(w) 0.967. This result suggests that during growth under these conditions, E. coli, although able to transcribe the required mRNA, may lack the cellular resources required for translation. Elucidating the global adaptive response of E. coli O157:H7 during exposure to chilling and water activity stress has provided a baseline of knowledge of the physiology of this pathogen.

A survey of Australian oysters for the presence of human noroviruses.

Impending international policies for norovirus in oysters and the lack of Australian data suggested there was a need to undertake a national survey of norovirus in oysters. Two geographically distinct oyster-growing areas from each of three Australian states were sampled on 4 occasions during 2010 and 2011. The sites selected were considered by state shellfish authorities to be the most compromised with respect to the potential for human faecal contamination as identified by shoreline surveys. The oysters were tested for norovirus GI, GII and Escherichia coli. Norovirus GII was detected in two of 120 (1.7%) samples and norovirus GI was not detected. One of the norovirus positive samples was cloned and sequenced as GII.3. Five of 120 (4.2%) samples were found to have more than the guidance concentration of 230 E. coli per 100 g of shellfish but these samples did not contain detectable concentrations of norovirus. The apparently low prevalence of norovirus in oysters from Australian growing areas supports epidemiological data that suggests norovirus contamination of Australian oysters is rare. The results from this study emphasise the need for future norovirus control measures for shellfish to be commensurate with the risk associated with the growing area.

Development of a microbial time-temperature indicator for real-time monitoring the quality of Australian vacuum-packed lamb.

A time-temperature indicator (TTI) system based on the pH-dependent colour change caused by the growth of a Carnobacterium maltaromaticum strain was developed to specifically provide a real-time indication of quality and shelf life of Australian vacuum-packed (VP) lamb throughout cold chains. Each component of the developed TTI system was studied to select an optimal concentration of a chemical chromatic indicator (chlorophenol red, CR; between 0.01 % and 0.30 %) and supplementary glucose (between 0 % and 10 %), and an appropriate C. maltaromaticum strain (among four different strains) in a simple BHI medium. BHI medium containing 0.01 % CR and 1 % added glucose, inoculated with C. maltaromaticum strain 1 were required for development of the TTI system to indicate quality and shelf life of VP lamb. Different inoculum levels of C. maltaromaticum strain 1 (103 to 105 CFU/mL) were also examined at 8 °C for their effects on the TTI response. As expected, higher inoculum levels of C. maltaromaticum led to a shorter endpoint of the TTI system but it was found that a 3 log10 higher inoculum level in the TTI than the expected total viable counts of VP lamb was required to accurately predict VP lamb shelf life by the TTI. To further evaluate the applicability of the TTI system, we evaluated its response at two other temperatures (2 °C and 4 °C) relevant to the storage conditions for VP lamb. The data showed a strong agreement between the observed TTI's endpoints and predicted shelf lives of VP lamb. This indicated that the developed TTI has the potential to be developed further for commercial application to provide a real-time, distinct, and accurate indication of Australian VP lamb.

Effect of pH, salt and chemical rinses on bacterial attachment to extracellular matrix proteins.

Microbial contamination of carcass surfaces occurs during slaughter and post-slaughter processing steps, therefore interventions are needed to enhance meat safety and quality. Although many studies have been done at the macro-level, little is known about specific processes that influence bacterial attachment to carcass surfaces, particularly the role of extracellular matrix (ECM) proteins. In the present study, the effect of pH and salt (NaCl, KCl and CaCl2) on attachment of Escherichia coli and Salmonella isolates to dominant ECM proteins: collagen I, fibronectin, collagen IV and laminin were assessed. Also, the effects of three chemical rinses commonly used in abattoirs (2% acetic acid, 2% lactic acid and 10% trisodium phosphate (TSP)) were tested. Within a pH range of 5-9, there was no significant effect on attachment to ECM proteins, whereas the effect of salt type and concentration varied depending on combination of strain and ECM protein. A concentration-dependant effect was observed with NaCl and KCl (0.1-0.85%) on attachment of E. coli M23Sr, but only to collagen I. One-tenth percent CaCl2 produced the highest level of attachment to ECM proteins for E. coli M23Sr and EC614. In contrast, higher concentrations of CaCl2 increased attachment of E. coli EC473 to collagen IV. Rinses containing TSP produced >95% reduction in attachment to all ECM proteins. These observations will assist in the design of targeted interventions to prevent or disrupt contamination of meat surfaces, thus improving meat safety and quality.

Understanding the microbial fibre degrading communities & processes in the equine gut.

The equine gastrointestinal tract is a self-sufficient fermentation system, housing a complex microbial consortium that acts synergistically and independently to break down complex lignocellulolytic material that enters the equine gut. Despite being strict herbivores, equids such as horses and zebras lack the diversity of enzymes needed to completely break down plant tissue, instead relying on their resident microbes to carry out fibrolysis to yield vital energy sources such as short chain fatty acids. The bulk of equine digestion occurs in the large intestine, where digesta is fermented for 36-48 h through the synergistic activities of bacteria, fungi, and methanogenic archaea. Anaerobic gut dwelling bacteria and fungi break down complex plant polysaccharides through combined mechanical and enzymatic strategies, and notably possess some of the greatest diversity and repertoire of carbohydrate active enzymes among characterized microbes. In addition to the production of enzymes, some equid-isolated anaerobic fungi and bacteria have been shown to possess cellulosomes, powerful multi-enzyme complexes that further enhance break down. The activities of both anaerobic fungi and bacteria are further facilitated by facultatively aerobic yeasts and methanogenic archaea, who maintain an optimal environment for fibrolytic organisms, ultimately leading to increased fibrolytic microbial counts and heightened enzymatic activity. The unique interactions within the equine gut as well as the novel species and powerful mechanisms employed by these microbes makes the equine gut a valuable ecosystem to study fibrolytic functions within complex communities. This review outlines the primary taxa involved in fibre break down within the equine gut and further illuminates the enzymatic strategies and metabolic pathways used by these microbes. We discuss current methods used in analysing fibrolytic functions in complex microbial communities and propose a shift towards the development of functional assays to deepen our understanding of this unique ecosystem.

Long-read MinION™ sequencing of 16S and 16S-ITS-23S rRNA genes provides species-level resolution of Lactobacillaceae in mixed communities.

The Lactobacillaceae are lactic acid bacteria harnessed to deliver important outcomes across numerous industries, and their unambiguous, species-level identification from mixed community environments is an important endeavor. Amplicon-based metataxonomics using short-read sequencing of partial 16S rRNA gene regions is widely used to support this, however, the high genetic similarity among Lactobacillaceae species restricts our ability to confidently describe these communities even at genus level. Long-read sequencing (LRS) of the whole 16S rRNA gene or the near complete rRNA operon (16S-ITS-23S) has the potential to improve this. We explored species ambiguity amongst Lactobacillaceae using in-silico tool RibDif2, which identified allele overlap when various partial and complete 16S rRNA gene and 16S-ITS-23S rRNA regions were amplified. We subsequently implemented LRS by MinION™ to compare the capacity of V3-V4, 16S and 16S-ITS-23S rRNA amplicons to accurately describe the diversity of a 20-species Lactobacillaceae mock community in practice. In-silico analysis identified more instances of allele/species overlap with V3-V4 amplicons (n = 43) compared to the 16S rRNA gene (n = 11) and partial (n = up to 15) or complete (n = 0) 16S-ITS-23S rRNA amplicons. With subsequent LRS of a DNA mock community, 80% of target species were identified using V3-V4 amplicons whilst the 16S rRNA gene and 16S-ITS-23S rRNA region amplicons resulted in 95 and 100% of target species being identified. A considerable reduction in false-positive identifications was also seen with 16S rRNA gene (n = 3) and 16S-ITS-23S rRNA region (n = 9) amplicons compared with V3-V4 amplicons (n = 43). Whilst the target species affected by allele overlap in V3-V4 and 16S rRNA gene sequenced mock communities were predicted by RibDif2, unpredicted species ambiguity was observed in 16S-ITS-23S rRNA sequenced communities. Considering the average nucleotide identity (ANI) between ambiguous species (~97%) and the basecall accuracy of our MinION™ sequencing protocol (96.4%), the misassignment of reads between closely related taxa is to be expected. With basecall accuracy exceeding 99% for recent MinION™ releases, the increased species-level differentiating power promised by longer amplicons like the 16S-ITS-23S rRNA region, may soon be fully realized.

Investigation of the Listeria monocytogenes Scott A acid tolerance response and associated physiological and phenotypic features via whole proteome analysis.

The global proteomic responses of the foodborne pathogen Listeria monocytogenes strain Scott A, during active growth and transition to the stationary growth phase under progressively more acidic conditions, created by addition of lactic acid and HCl, were investigated using label-free liquid chromatography/tandem mass spectrometry. Approximately 56% of the Scott A proteome was quantitatively assessable, and the data provides insight into its acquired acid tolerance response (ATR) as well as the relation of the ATR to the growth phase transition. Alterations in protein abundance due to acid stress were focused in proteins belonging to the L. monocytogenes common genome, with few strain-dependent proteins involved. However, one of the two complete prophage genomes appeared to enter lysogeny. During progressive acidification, the growth rate and yield were reduced 55% and 98%, respectively, in comparison to nonacidified control cultures. The maintenance of the growth rate was determined to be connected to activation of cytoplasmic pH homeostatic mechanisms while cellular reproductive-related and cell component turnover proteins were markedly more abundant in acid stressed cultures. Cell biomass accumulation was impeded predominantly due to repression of phosphodonor-linked enzymes involved with sugar phosphotransfer, glycolysis, and cell wall polymer biosynthesis. Acidification caused a shift from heterofermentation to an oxidatively stressed state in which ATP appears to be generated mainly through the pyruvate dehydrogenase/pyruvate oxidase/phosphotransacetylase/acetate kinase and branched chain acid dehydrogenase pathways. Analysis of regulons indicated energy conservation occurs due to repression by the GTP/isoleucine sensor CodY and also the RelA mediated stringent response. Whole proteome analysis proved to be an effective way to highlight proteins involved with the acquisition of the ATR.

MudPIT analysis of alkaline tolerance by Listeria monocytogenes strains recovered as persistent food factory contaminants.

Alkaline solutions are used to clean food production environments but the role of alkaline resistance in persistent food factory contamination by Listeria monocytogenes is unknown. We used shotgun proteomics to characterise alkaline adapted L. monocytogenes recovered as persistent and transient food factory contaminants. Three unrelated strains were studied including two persistent and a transient food factory contaminant determined using multilocus sequence typing (MLST). The strains were adapted to growth at pH 8.5 and harvested in exponential phase. Protein extracts were analysed using multidimensional protein identification technology (MudPIT) and protein abundance compared by spectra counting. The strains elicited core responses to alkaline growth including modulation of intracellular pH, stabilisation of cellular processes and reduced cell-division, independent to lineage, MLST or whether the strains were transient or persistent contaminants. Alkaline adaptation by all strains corresponded to that expected in stringent-response induced cells, with protein expression supporting metabolic shifts concordant with elevated alarmone production and indicating that the alkaline-stringent response results from energy rather than nutrient limitation. We believe this is the first report describing induction of a stringent response in different L. monocytogenes strains by alkaline pH under non-limiting growth conditions. The work emphasises the need for early intervention to avoid persistent food factory contamination by L. monocytogenes.

Modelling growth and histamine formation of Klebsiella aerogenes TI24 isolated from Indonesian pindang.

Indonesian salted-boiled fish (pindang) is a popular traditional food in Indonesia, which is made from Scombroid fish such as tuna and mackerel. As with other traditionally prepared fish products, pindang has important economic and social values, especially for those living in the coastal areas of Indonesia. However, pindang is a major cause of histamine fish poisoning (HFP) for consumers. Klebsiella aerogenes T124, a relatively high histamine-producing isolate from pindang, was used to describe lag time (λ), growth rate (µmax), maximum population density (Nmax), and histamine production in histidine broth and artificially contaminated Grey mackerel. Broth was adjusted to 1.5, 6, 10 and 20% w/v NaCl; mackerel was treated with 6% w/w NaCl, a level common to Indonesian industry practice, or not treated with additional NaCl. Samples were incubated at 10, 15, 20 and 30 °C. In broth, µmax and Nmax were significantly affected by temperature and NaCl, respectively, with λ influenced by both parameters. In control fish, µmax was significantly affected by temperature and NaCl, except at 10 and 15 °C; for 6% NaCl treatment, growth was only observed at 20 and 30 °C. Under similar incubation conditions for broth and fish, histamine formation was markedly affected by NaCl concentration. In broth, -5.1 to -6.6 log µg of histamine was produced per CFU, versus -4.6 to -6.6 log µg per CFU in fish. This study demonstrated that mackerel treated with 6% NaCl and stored at 10-15 °C prevents growth of K. aerogenes strain TI24 and formation of toxic levels of histamine.

The effects of glucose on microbial spoilage of vacuum-packed lamb.

Vacuum-packed lamb produced in Australia has a shelf-life of 80-90 days under export conditions (-1 to 0 °C). However, access to some markets could involve >90 days transit time. Studies to understand the potential mechanisms of microbial spoilage of vacuum-packed lamb are, therefore, important to assist the development of shelf-life extension methods. Here, we investigated the effects of glucose on the shelf-life of vacuum-packed lamb. This was done by adding glucose (up to 4.64 mmol/kg) to the surface of meat and conducting a series of shelf-life trials, in which the sensorial qualities, bacterial growth, pH, and residual glucose and lactic acid were measured over time. Based on sensory analysis glucose extended the shelf-life, ranging from 8% to >76% increase relative to the control. Glucose reduced meat pH, potentially affecting the microbial community composition and the accumulation of spoilage metabolites. These results indicate that glucose plays an important role in microbial spoilage of vacuum-packed lamb possibly by pH reduction.

Predictive models for the effect of storage temperature on Vibrio parahaemolyticus viability and counts of total viable bacteria in Pacific oysters (Crassostrea gigas).

Vibrio parahaemolyticus is an indigenous bacterium of marine environments. It accumulates in oysters and may reach levels that cause human illness when postharvest temperatures are not properly controlled and oysters are consumed raw or undercooked. Predictive models were produced by injecting Pacific oysters (Crassostrea gigas) with a cocktail of V. parahaemolyticus strains, measuring viability rates at storage temperatures from 3.6 to 30.4°C, and fitting the data to a model to obtain parameter estimates. The models were evaluated with Pacific and Sydney Rock oysters (Saccostrea glomerata) containing natural populations of V. parahaemolyticus. V. parahaemolyticus viability was measured by direct plating samples on thiosulfate-citrate-bile salts-sucrose (TCBS) agar for injected oysters and by most probable number (MPN)-PCR for oysters containing natural populations. In parallel, total viable bacterial counts (TVC) were measured by direct plating on marine agar. Growth/inactivation rates for V. parahaemolyticus were -0.006, -0.004, -0.005, -0.003, 0.030, 0.075, 0.095, and 0.282 log10 CFU/h at 3.6, 6.2, 9.6, 12.6, 18.4, 20.0, 25.7, and 30.4°C, respectively. The growth rates for TVC were 0.015, 0.023, 0.016, 0.048, 0.055, 0.071, 0.133, and 0.135 log10 CFU/h at 3.6, 6.2, 9.3, 14.9, 18.4, 20.0, 25.7, and 30.4°C, respectively. Square root and Arrhenius-type secondary models were generated for V. parahaemolyticus growth and inactivation kinetic data, respectively. A square root model was produced for TVC growth. Evaluation studies showed that predictive growth for V. parahaemolyticus and TVC were "fail safe." The models can assist oyster companies and regulators in implementing management strategies to minimize V. parahaemolyticus risk and enhancing product quality in supply chains.

Utility of gel-free, label-free shotgun proteomics approaches to investigate microorganisms.

This review will examine the current situation with label-free, quantitative, shotgun-oriented proteomics technology and discuss the advantages and limitations associated with its capability in capturing and quantifying large portions of proteomes of microorganisms. Such an approach allows (1) comparisons between physiological or genetic states of organisms at the protein level, (2) 'painting' of proteomic data onto genome data-based metabolic maps, (3) enhancement of the utility of genomic data and finally (4) surveying of non-genome sequenced microorganisms by taking advantage of available inferred protein data in order to gain new insights into strain-dependent metabolic or physiological capacities. The technology essentially is a powerful addition to systems biology with a capacity to be used to ask hypothesis-driven 'top-down' questions or for more empirical 'bottom-up' exploration.