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.

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.

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.

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.

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.

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.

Big Data Impacting Dynamic Food Safety Risk Management in the Food Chain.

Foodborne pathogens are a major contributor to foodborne illness worldwide. The adaptation of a more quantitative risk-based approach, with metrics such as Food safety Objectives (FSO) and Performance Objectives (PO) necessitates quantitative inputs from all stages of the food value chain. The potential exists for utilization of big data, generated through digital transformational technologies, as inputs to a dynamic risk management concept for food safety microbiology. The industrial revolution in Internet of Things (IoT) will leverage data inputs from precision agriculture, connected factories/logistics, precision healthcare, and precision food safety, to improve the dynamism of microbial risk management. Furthermore, interconnectivity of public health databases, social media, and e-commerce tools as well as technologies such as blockchain will enhance traceability for retrospective and real-time management of foodborne cases. Despite the enormous potential of data volume and velocity, some challenges remain, including data ownership, interoperability, and accessibility. This paper gives insight to the prospective use of big data for dynamic risk management from a microbiological safety perspective in the context of the International Commission on Microbiological Specifications for Foods (ICMSF) conceptual equation, and describes examples of how a dynamic risk management system (DRMS) could be used in real-time to identify hazards and control Shiga toxin-producing Escherichia coli risks related to leafy greens.

Effect of peroxyacetic acid treatment and bruising on the bacterial community and shelf-life of baby spinach.

The importance of leaf integrity, i.e. the effects of bruising (mechanical damage), and sanitisation with peroxyacetic acid (PAA) on bacterial communities of ready-to-eat baby spinach remains unclear. Two shelf-life studies were conducted at 4 °C to investigate the effect of bruising and sanitisation on the growth of spoilage microorganisms. In the first experiment, both bruising treatments (100% and 40% of leaves) halved shelf life to 12 d, whereas intact leaves had a shelf-life of 23 d. Bruising had no influence on bacterial diversity during shelf-life, though some differences in the relative abundance of minor genera were observed. Pseudomonas and Pantoea were the most dominant bacterial genera, regardless of bruising treatment. High throughput amplicon sequencing also identified other spoilage bacteria including Chryseobacterium, Stenotrophomonas, Bacillus, Sphingobacterium, Erwinia and Flavobacterium. In the second experiment, washing of intact baby spinach with a sanitiser (80 mg/L: PAA) reduced microbial load as determined by aerobic plate count but did not immediately affect the presence/relative abundance of most of the genera of spoilage bacteria observed. During shelf-life, the bacterial diversity of sanitised leaves was significantly lower than on water-washed leaves. Although sanitisation resulted in a higher initial log reduction in microbial load compared to control (portable tap water), sanitisation did not extend the shelf life of baby spinach (23 d). Sanitised spinach had reduced bacterial diversity however, by the end of shelf life, both sanitised and water-washed spinach was dominated by Pseudomonas and Pantoea spoilage bacteria. This study demonstrated for the first time that the shorter shelf life of bruised leaves was related to faster microbial growth rather than changes in bacterial diversity or community composition.

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.

Effect of Storage Conditions on Shelf Stability of Undiluted Neutral Electrolyzed Water.

ABSTRACT: Neutral electrolyzed water (NEW) is an oxidizing sanitizer that can be made locally on-site; it is often stored in a ready-to-use format to accumulate the large volumes required for periodic or seasonal use. The shelf stability of NEW sanitizer was, therefore, assessed under various storage conditions to guide the development of protocols for its industrial application. To that end, fresh NEW with an available chlorine concentration (ACC) of 480 mg/L, pH 6.96, and oxidation reduction potential (ORP) of 916 mV was stored under different conditions. These were open or sealed polypropylene bottles, three different surface area-to-volume (SA:V) ratios (0.9, 1.7, and 8.7), and two temperatures (4 and 25°C). NEW stored at 4°C was significantly more stable than NEW stored at 25°C; ACC and pH decreased by 137 mg/L and 0.7, respectively, whereas ORP increased by 23 mV, after 101 days of storage. At 25°C, ACC decreased to <0.01 mg/L after 52 days in bottles with a SA:V ratio of 8.7, with a similar decrease after 101 days in bottles with a SA:V ratio of 1.7. However, pH decreased by up to 3.7 pH units, and ORP increased by up to 208 mV. The antimicrobial efficacy of "aged" electrolyzed oxidizing (EO) water with different ACC and ORP, but the same pH (i.e., 3.4 ± 0.2), was evaluated against Escherichia coli and Listeria innocua to determine any differences in residual antimicrobial activity. EO water with an ACC of ≥7 mg/L and an ORP of 1,094 mV caused a reduction of at least 4.7 log, whereas EO water with nondetectable ACC and considerably high ORP (716 mV) had little antimicrobial effect (<1-log reduction). Results from this study indicate that the efficacy of NEW as a sanitizer for large-scale applications such as horticulture can be maintained for at least 3 months when it is stored in closed containers with low SA:V ratio at low temperatures.

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.

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.

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.

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.

Automated Fall Detection Technology in Inpatient Geriatric Psychiatry: Nurses' Perceptions and Lessons Learned.

ABSTRACTHospitalized older adults are at high risk of falling. The HELPER system is a ceiling-mounted fall detection system that sends an alert to a smartphone when a fall is detected. This article describes the performance of the HELPER system, which was pilot tested in a geriatric mental health hospital. The system's accuracy in detecting falls was measured against the hospital records documenting falls. Following the pilot test, nurses were interviewed regarding their perceptions of this technology. In this study, the HELPER system missed one documented fall but detected four falls that were not documented. Although sensitivity (.80) of the system was high, numerous false alarms brought down positive predictive value (.01). Interviews with nurses provided valuable insights based on the operation of the technology in a real environment; these and other lessons learned will be particularly valuable to engineers developing this and other health and social care technologies.

Spatial and Temporal Distribution of Norovirus and E. coli in Sydney Rock Oysters Following a Sewage Overflow into an Estuary.

This paper reports a study of norovirus (NoV) GII distribution and persistence in Sydney rock oysters (SRO) (Saccostrea glomerata) located in an estuary after a pump station sewage overflow. SRO were strategically placed at six sites spanning the length of the estuary from the pump station to the sea. The spatial and temporal distribution of NoV, hepatitis A virus (HAV) and Escherichia coli (E. coli) in oysters was mapped after the contamination event. NoV GI and GII, HAV and E. coli were quantified for up to 48 days in oysters placed at six sites ranging from 0.05 to 8.20 km from the sewage overflow. NoV GII was detected up to 5.29 km downstream and persisted in oysters for 42 days at the site closest to the overflow. NoV GII concentrations decreased significantly over time; a reduction rate of 8.5% per day was observed in oysters (p < 0.001). NoV GII concentrations decreased significantly as a function of distance at a rate of 5.8% per km (p < 0.001) and the decline in E. coli concentration with distance was 20.1% per km (p < 0.001). HAV and NoV GI were not detected. A comparison of NoV GII reduction rates from oysters over time, as observed in this study and other published research, collectively suggest that GII reduction rates from oysters may be broadly similar, regardless of environmental conditions, oyster species and genotype.

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.

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.

Correction: The Biokinetic Spectrum for Temperature.

[This corrects the article DOI: 10.1371/journal.pone.0153343.].

The Biokinetic Spectrum for Temperature.

We identify and describe the distribution of temperature-dependent specific growth rates for life on Earth, which we term the biokinetic spectrum for temperature. The spectrum has the potential to provide for more robust modeling in thermal ecology since any conclusions derived from it will be based on observed data rather than using theoretical assumptions. It may also provide constraints for systems biology model predictions and provide insights in physiology. The spectrum has a Δ-shape with a sharp peak at around 42°C. At higher temperatures up to 60°C there was a gap of attenuated growth rates. We found another peak at 67°C and a steady decline in maximum rates thereafter. By using Bayesian quantile regression to summarise and explore the data we were able to conclude that the gap represented an actual biological transition between mesophiles and thermophiles that we term the Mesophile-Thermophile Gap (MTG). We have not identified any organism that grows above the maximum rate of the spectrum. We used a thermodynamic model to recover the Δ-shape, suggesting that the growth rate limits arise from a trade-off between activity and stability of proteins. The spectrum provides underpinning principles that will find utility in models concerned with the thermal responses of biological processes.