Strain variability in growth and thermal inactivation characteristics of Listeria monocytogenes strains after acid adaptation

Given the importance of strain variability to predictive microbiology and risk assessment, the present study aimed to quantify the magnitude of strain variability in growth and thermal inactivation kinetics behaviors after acid adaptation. Thirty-three Listeria monocytogenes strains were exposed to acid-adapted tryptic soy broth with yeast extract and nonacid-adapted TSB-YE (pH 7.0) for 20 hours. Then, the growth parameters of these adapted and non-adapted strains that grew in non-buffered TSB-YE at 25℃ were estimated. The tested strains were inactivated at 60°C in non-buffered broth to obtain the heat resistance parameters. The results revealed that strain variability was present in the growth and thermal inactivation characteristics. The maximum specific growth rate ( μ max ) ranged within 0.21-0.44 and 0.20-0.45 h -1 after acid and non-acid adaptation, respectively. The lag times ( λ ) were 0.69-2.56 and 0.24-3.36 hours for acid-adapted and non-acid adapted cells, respectively. The apparent D -values at 60°C ( D 60 -values) of the pathogen ranged within 0.56-3.93 and 0.52-3.63 minutes for the presence and absence of acid adaptation condition, respectively. Acid adaptation increased the magnitude of strain variability in the thermal inactivation characteristics of the organism ( P <0.05), with the coefficient of variation (CV) increasing to 0.17, while acid adaptation did not significantly influence the variabilities in the growth parameters of the tested strains ( P ≥0.05). Furthermore, the subsequent growth behaviors of all strains did not exhibit significant changes ( P >0.05) after exposure to acidic broth. However, the thermal resistance of most of the tested strains (25/33) increased ( P <0.05) after growing in acid-adapted broth. The relevant data generated in the present study can be used to describe the strain variability in predictive microbiology, and deeply understand the behavior responses of different strains to acid adaptation.

Cancer Cell Acid Adaptation Gene Expression Response Is Correlated to Tumor-Specific Tissue Expression Profiles and Patient Survival

The acidic pH of the tumor microenvironment plays a critical role in driving cancer development toward a more aggressive phenotype, but the underlying mechanisms are unclear. To this end, phenotypic and genotypic changes induced by adaptation of cancer cells to chronic acidosis have been studied. However, the generality of acid adaptation patterns across cell models and their correlation to the molecular phenotypes and aggressiveness of human cancers are essentially unknown. Here, we define an acid adaptation expression response shared across three cancer cell models, dominated by metabolic rewiring, extracellular matrix remodeling, and altered cell cycle regulation and DNA damage response. We find that many genes which are upregulated by acid adaptation are significantly correlated to patient survival, and more generally, that there are clear correlations between acid adaptation expression response and gene expression change between normal and tumor tissues, for a large subset of cancer patients. Our data support the notion that tumor microenvironment acidity is one of the key factors driving the selection of aggressive cancer cells in human patient tumors, yet it also induces a growth-limiting genotype that likely limits cancer cell growth until the cells are released from acidosis, for instance during invasion.

Effect of Ph On Survival of Escherichia coli O157, Escherichia coli O121, and Salmonella enterica During Desiccation and Short-term Storage

ABSTRACT One intrinsic characteristic of low-moisture foods that is frequently overlooked is pH. Although pH affects the survival of microorganisms in high-moisture foods, its influence in low-moisture foods with less available moisture has not been examined. Escherichia coli O157:H7, E. coli O121, Salmonella enterica Anatum, and S. enterica Agona were grown on solid media with and without added glucose, harvested, and then suspended in buffer at pH 4, 5, and 7 for 10 min. All cultures were spotted individually onto cellulose filters and dried in a biohazard cabinet (23 ± 2°C) overnight (24 ± 2 h) and then stored in a 25°C incubator at 33% relative humidity. Populations were examined at regular intervals up to 26 (E. coli) or 29 (Salmonella) days. Additional controls for pH consisted of cultures held in buffer at pH 4, 5, and 7 at 25°C for the same time periods as the desiccated cells. For all strains tested, pH had an effect on survival whether stored dried or in liquid buffer (P < 0.05). However, when grown on solid media, acid adaptation (grown with glucose) before acid treatment did not appear beneficial to Salmonella during desiccation. Instead, both acid-adapted Salmonella serovars appeared less resistant during drying than did non–acid-adapted cells. Once dried, the rates of decline for Salmonella were not significantly different for acid-adapted and nonadapted cells (P > 0.05), indicating similar persistence following desiccation. A reverse trend was observed for E. coli O121; acid adaptation on solid media improved survival during desiccation and subsequent storage at low pH (P < 0.05). E. coli O157:H7 survival was significantly lower than that of either Salmonella or E. coli O121 under all conditions tested. Results indicate that the response to desiccation and pH stress differs between the microorganisms and under different growth conditions. HIGHLIGHTS

Collagen Production and Niche Engineering: A Novel Strategy for Cancer Cells to Survive Acidosis and Evolve

SummaryDuctal Carcinoma in situ (DCIS) is an avascular disease characterized by profound acidosis. Pre-malignant cells within this niche must adapt to acidosis to survive and thrive. A component of this acid-adaptation involves extracellular matrix remodeling leading to niche construction and remodeling. Using discovery proteomics, we identified that collagen producing enzyme PLODs are upregulated in acid-adapted breast cancer cells. Second harmonic generation microscopy showed significant collagen deposition within DCIS lesions of patients. In vitro analyses identified that acid-adaptation involves production of rare collagens that can be regulated by Ras and SMAD pathway. Secretome analysis showed upregulation ECM remodeling enzymes such as TGM2 and LOXL2. Comparison of acid induced collagens in vitro and in patient data showed correlation between rare collagens production and survival of patients. We conclude acidosis induces collagen production by cancer cells and promote growth independent of basal membrane attachment. The independently produced collagen can be used for niche construction and engineering as an adaptation strategy of cancer cells to survive and evolve.

Aside Adapte ve Adapte Olmayan Durağan Faz Escherichia coli O157:H7, Salmonella enterica Typhimurium ve Listeria monocytogenes’in Nar Suyundaki Termal Direnci

The purpose of this study was to investigate the thermal resistance of acid adapted and non-adapted stationary phase Escherichia coli O157:H7, Salmonella enterica Typhimurium and Listeria monocytogenes in pomegranate juice. In addition, the performance of generic E. coli was evaluated as an indicator. Non-adapted stationary phase cells were grown by incubating inoculated tryptic soy broth without glucose (TSB-NG) at 36±1°C for 18±2 hours. Tryptic soy broth with 1% glucose (10 g/l; TSBG) was used for acid adaptation. All media used for L. monocytogenes was supplemented with 0.6% yeast extract. After washing the cells with peptone, 5 ml of pasteurized pomegranate juice was added onto the pellet to obtain inoculated juice with a initial concentration of 107-1010 log CFU/ml. Inoculated pomegranate juice was sealed into the microcapillary tubes. Microtubes were heat treated in waterbaths at 50, 52 and 54±1°C by immersing at pre-determined time intervals. Survived populations were counted on tryptic soy agar (TSA). S. Typhimurium had the lowest thermal resistance in pomegranate juice. At 50°C, E. coli O157:H7 was the most resistant, whereas L. monocytogenes was more thermally tolerant at 52 and 54°C. Acid adaptation decreased the thermal resistance of E. coli O157:H7, but increased the heat resistance of L. monocytogenes at all tested temperatures significantly. Thermal tolerance of S. Typhimurium increased only at 50°C. The most resistant microorganism was non-adapted generic E. coli at 50 and 52°C; acid-adapted L. monocytogenes had the most thermal tolerance at 54°C. Thermal inactivation of microorganisms in pomegranate juice could be tested at lower temperatures compare to other fruit juices. This may be due to the natural antimicrobial effect and more acidic content of pomegranate juice.