Experimentally observed Campylobacter jejuni survival kinetics in chicken meat products during model gastric digestion tended to be lower than model predictions.

Campylobacter jejuni-related foodborne diseases are mainly attributed to the consumption of undercooked chicken meat and cross-contaminated produce. This study aimed to develop a survival kinetics model, based on the Weibull model, for predicting foodborne C. jejuni survival during gastric digestion in a model stomach. We previously confirmed that C. jejuni can survive temperatures up to 62 °C; therefore, certain types of grilled chicken skewers (yakitori) were examined for C. jejuni survival during simulated gastric digestion. C. jejuni survival on a chicken thigh following grilling was examined to confirm the foods for digestion experiments. Further, C. jejuni survival during model digestion was investigated through simultaneous digestion of raw chicken and cross-contaminated iceberg lettuce. The model stomach pH increased from 1.5 to 6.0 immediately after yakitori ingestion and did not decrease below 4.0 within 3 h of digestion. Gastric digestion did not significantly contribute to C. jejuni inactivation (<1.5 log reduction after 3 h digestion). Our model could predict C. jejuni survival kinetics in simulated gastric fluid under varying pH during model digestion. This approach can be used to predict C. jejuni survival rates following digestion to improve food safety and reduce Campylobacter-related disease outbreaks.

Listeria monocytogenes survives better at lower storage temperatures in regular and low-salt soft and cured cheeses.

The behaviour of Listeria monocytogenes was investigated in soft pasteurized milk cheese elaborated with different salt concentrations (1.17 and 0.30% w/w) and in cured raw sheep milk cheese over storage up to 189 days at different isothermal conditions. Commercial 25-g cheese samples were inoculated with a 4-strain cocktail of L. monocytogenes (serovars 4b, 1/2a, 1/2b and 1/2c) at approximately 104 CFU/g. The inoculated samples were stored at 4 and 22 °C and withdrawn at proper intervals for L. monocytogenes enumeration. The prevalence of the different serovar strains of L. monocytogenes was characterized on soft cheese samples over storage at 4 °C using multiplex PCR. Salt reduction did not affect the survival of L. monocytogenes in soft cheeses and a maximum of 1-log reduction was observed in both regular and low-salt cheeses after 189 days of storage at 4 °C. The pathogen showed greater survival capacity in both soft and cured cheeses during storage at 4 °C compared to the storage at 22 °C, where more than 2.5 log reductions were computed. The fate of L. monocytogenes was described through a Weibull model fitted to survival data. The time required for a first tenfold reduction of the L. monocytogenes population (δ) at 4 °C is around 150 days in soft and 72 days in cured cheeses. At 22 °C, the estimated δ values are at least 60% lower in both cheese types. Among the four L. monocytogenes serovars present in the inoculated cocktail, the serovar 4b strain was the most sensitive to refrigerated storage, while the prevalence of serovar 1/2c strain increased over time in soft cheeses. Overall, the data obtained in this study help to deepen knowledge into factors affecting L. monocytogenes behaviour on cheeses and evidenced the variability between serovars in terms of survival capacity, which may be considered when performing microbial risk assessments.

Survival of Clostridioides difficile in finished dairy compost under controlled conditions.

AIM: The survival of Clostridioides difficile (previously Clostridium difficile) vegetative cells and endospores was compared at different levels of indigenous microflora using autoclaved and unautoclaved dairy composts with different moisture contents (MCs). METHODS AND RESULTS: Both types of composts adjusted to 20, 30 and 40% MCs were inoculated with a suspension of C. difficile that contained both vegetative cells (c. 5-6 log CFU per gram) and endospores (c. 5·0 CFU per gram), and then stored aerobically inside a humidity-controlled chamber at room temperature 22·5 ± 0·8°C for 1 year. The level of indigenous microflora was very stable during the storage after day 7 in both types of compost. The greatest reductions of C. difficile vegetative cell counts occurred during the first 24 h of storage in autoclaved and unautoclaved composts, which had 4·7 and 5·5 log CFU per gram with 20% MC, 1·8 and 2·1 log CFU per gram with 30% MC, and 2·3 and 1·3 log CFU per gram with 40% MC, respectively. Both MC and the duration of storage have significant (P < 0·05) effects on the survival of vegetative cells for first 120 days of storage. The slow inactivation of C. difficile vegetative cells at higher MCs during aerobic storage was confirmed by exponentially decaying modelling data during the early stage of aerobic exposure. The reduction of endospore counts (<1·0 log CFU per gram) during the storage for both types of compost at all MCs was not significant (P > 0·05) except for the autoclaved compost with 30% MC. CONCLUSION: The highly resistant C. difficile endospores to the unfavourable environmental conditions survived for more than a year while vegetative cells died off exponentially upon the initial aerobic exposure. SIGNIFICANCE AND IMPACT OF THE STUDY: The long-term survival of C. difficile endospores in contaminated compost may transmit the pathogen to fresh produce, animals or water in pre-harvest conditions.

Influence of application sequence and timing of eugenol and lauric arginate (LAE) on survival of spoilage organisms.

The effectiveness of sequential applications of the antimicrobials eugenol and lauric arginate (LAE) was investigated against Staphylococcus carnosus, Listeria innocua, Escherichia coli K12, and Pseudomonas fluorescens. The antimicrobials were applied simultaneously at half of their minimum lethal concentrations (MLC) or sequentially at t = 0 h and t = 3, 4, 6 or 8 h. Bacterial survival was determined by direct plate counts. Survivals kinetic were fitted to a growth and mortality model to obtain characteristic parameters that described time-dependent changes from growth to mortality or vice versa. The most effective was a simultaneous exposure of both antimicrobials to the spoilage organisms at the beginning of the incubation period. Efficiency decreases depending on order and timing of the two antimicrobials were observed upon sequential treatments. These were most effective when antimicrobials where applied within a short time period (3-4 h) and when eugenol was first applied against S. carnosus and P. fluorescens. No sequence effects were observed for L. innocua, and sequential treatments proved to be ineffective against E. coli K12. These results were attributed to cells adapting to the first applied antimicrobial. In some cases, this provided protection against the second antimicrobial rendering the overall treatment less effective.

Survival kinetics of Mycobacterium bovis during manufacture and ripening of raw milk Cheddar and Caerphilly cheese produced on a laboratory-scale.

AIMS: Persistence of Mycobacterium bovis was investigated in UK raw milk cheeses. METHODS AND RESULTS: Replicating traditional cheese production methods under stringent CL3 containment conditions, Cheddar and Caerphilly cheeses were produced with Myco. bovis inoculated raw milk. High-inoculum investigations used three Myco. bovis genotypes; later low-inoculum investigations used only Myco. bovis AF2122/97. High-inoculum Cheddar (n = 9) and Caerphilly (n = 9) were matured for a minimum of 12 and 4 months respectively; maturation of low-inoculum Cheddar (n = 3) and Caerphilly (n = 3) was up to 11 weeks. Survival of Myco. bovis was monitored by enumeration at different points throughout cheese manufacture and ripening. D values were calculated as follows: 57 and 59 days in high-inoculum Cheddar and Caerphilly, respectively, and 41 and 24 days in low-inoculum Cheddar and Caerphilly respectively. CONCLUSIONS: Mycobacterium bovis is concentrated in cheese curd and a proportion lost with the whey. Reduction in viability during manufacturing is limited, while significant Myco. bovis inactivation occurs during maturation. Inactivation was improved, during Caerphilly ripening, when acid development was enhanced by increasing the proportion of starter culture. SIGNIFICANCE AND IMPACT OF THE STUDY: Mycobacterium bovis inactivation data obtained could be used to inform assessment of the risk posed to consumers by raw milk dairy products.

Population Survival Kinetics Derived from Clinical Trials of Potentially Curable Lung Cancers.

Using digitized data from progression-free survival (PFS) and overall survival Kaplan-Meier curves, one can assess population survival kinetics through exponential decay nonlinear regression analyses. To demonstrate their utility, we analyzed PFS curves from published curative-intent trials of non-small cell lung cancer (NSCLC) adjuvant chemotherapy, adjuvant osimertinib in resected EGFR-mutant NSCLC (ADAURA trial), chemoradiotherapy for inoperable NSCLC, and limited small cell lung cancer (SCLC). These analyses permit assessment of log-linear curve shape and estimation of the proportion of patients cured, PFS half-lives for subpopulations destined to eventually relapse, and probability of eventual relapse in patients remaining progression-free at different time points. The proportion of patients potentially cured was 41% for adjuvant controls, 58% with adjuvant chemotherapy, 17% for ADAURA controls, not assessable with adjuvant osimertinib, 15% with chemoradiotherapy, and 12% for SCLC. Median PFS half-life for relapsing subpopulations was 11.9 months for adjuvant controls, 17.4 months with adjuvant chemotherapy, 24.4 months for ADAURA controls, not assessable with osimertinib, 9.3 months with chemoradiotherapy, and 10.7 months for SCLC. For those remaining relapse-free at 2 and 5 years, the cure probability was 74%/96% for adjuvant controls, 77%/93% with adjuvant chemotherapy, 51%/94% with chemoradiation, and 39%/87% with limited SCLC. Relatively easy population kinetic analyses add useful information.

Kinetic and proteomic studies in milk show distinct patterns among major Listeria monocytogenes clones.

Listeria monocytogenes, a contaminant of raw milk, includes hypervirulent clonal complexes (CC) like CC1, CC4, and CC6, highly overrepresented in dairy products when compared to other food types. Whether their higher prevalence in dairy products is the consequence of a growth advantage in this food remains unknown. We examined growth kinetics of five L. monocytogenes isolates (CC1, CC4, CC6, CC9, and CC121) at 37 and 4 °C in ultra-high temperature (UHT) milk and raw milk. At 4 °C, hypovirulent CC9 and CC121 isolates exhibit better growth parameters in UHT milk compared to the hypervirulent CC1, CC4, and CC6 isolates. CC9 isolate in raw milk at 4 °C exhibited the fastest growth and the highest final concentrations. In contrast, hypervirulent isolates (CC1, CC4, and CC6) displayed better growth rates in UHT milk at 37 °C, the mammalian host temperature. Proteomic analysis of representative hyper- (CC1) and hypovirulent (CC9) isolates showed that they respond to milk cues differently with CC-specific traits. Proteins related to metabolism (such as LysA or different phosphotransferase systems), and stress response were upregulated in both isolates during growth in UHT milk. Our results show that there is a Listeria CC-specific and a Listeria CC-common response to the milk environment. These findings shed light on the overrepresentation of hypervirulent L. monocytogenes isolates in dairy products, suggesting that CC1 and CC4 overrepresentation in dairy products made of raw milk may arise from contamination during or after milking at the farm and discard an advantage of hypervirulent isolates in milk products when stored at refrigeration temperatures.

Comparison of inactivation kinetics of Yersinia enterocolitica in vegan and non-vegan kimchi during fermentation.

Yersinia enterocolitica is occasionally detected in kimchi, a traditional food prepared from fermented vegetables. Changes in growth properties of Y. enterocolitica during kimchi fermentation are largely unknown. We investigated the viability of Y. enterocolitica during the fermentation of vegan and non-vegan kimchi at different temperatures. Changes in Y. enterocolitica population, pH, and titratable acidity were measured for 24 days. In a suspension test with kimchi juice, populations of three Y. enterocolitica strains were above 3.30 log10 CFU/mL at pH > 5 for 7 days. Yersinia enterocolitica counts in vegan kimchi were considerably reduced at 0 °C and 6 °C. During fermentation at 6 °C, Y. enterocolitica populations in non-vegan kimchi and vegan kimchi were not detected starting from days 14 and 10, respectively. In kimchi samples stored at 0 °C and 6 °C, Y. enterocolitica survival correlated with pH changes during fermentation; in samples stored for up to 24 days, Y. enterocolitica was not detected. According to the k max values from the "log-linear with shoulder and tail" model, Y. enterocolitica was more sensitive to vegan kimchi fermentation than to non-vegan kimchi fermentation. Our findings provide an important basis for ensuring the safe production of kimchi without Y. enterocolitica contamination. Further research is necessary to elucidate the mechanism of Y. enterocolitica inactivation and the major bacterial and physicochemical factors involved in kimchi fermentation.

Effect of Different Pre-Growth Temperatures on the Survival Kinetics of Salmonella enterica and Listeria monocytogenes in Fresh-Cut Salad during Refrigerated Storage.

The effect of the pre-growth temperature of bacterial cultures on their subsequent survival kinetics in fresh-cut produce during refrigerated storage was investigated in this study. Three-strain cocktails of Listeria monocytogenes and Salmonella enterica, cultured at different growth temperatures (4, 21, and 37 °C) were inoculated on fresh-cut mixed salad and on individual produce in the mixed salad. The inoculated samples were stored at 4 °C and 80 ± 2% relative humidity (RH) for up to 72 h and the growth, survival, or death kinetics were determined at regular intervals. The results indicate that depending upon the type of pathogen tested, the pre-growth temperature(s) and the type of produce showed a significant (p ≤ 0.05) effect on the survival kinetics. Among the tested produce, mixed salad showed the highest reduction in L. monocytogenes pre-grown at 37 °C (1.33 log CFU/g) followed by red cabbage (0.56 log CFU/g), iceberg lettuce (0.52 log CFU/g), and carrot (-0.62 log CFU/g), after 72 h, respectively. In the case of Salmonella, carrot showed the highest reduction (1.07 log CFU/g for 37 °C pre-grown culture) followed by mixed salad (0.78 log CFU/g for 37 °C pre-grown culture), cabbage (0.76 log CFU/g for 21 °C pre-grown culture), and lettuce (0.65 log CFU/g for 4 °C pre-grown culture), respectively. Among the tested ComBase predictive models, the Baranyi-Roberts model better fitted the experimental data. These findings indicate that the appropriate selection of pre-growth environmental conditions is critical to better understand the kinetics of foodborne pathogens.

Survival of Staphylococcus aureus in dried fish products as a function of temperature.

We evaluated the survival ability of Staphylococcus aureus and the production of Staphylococcal enterotoxin A (SEA) in dried filefishes and julienned squid at 10°, 24°, and 35 °C for 5 months. S. aureus survived longer at 10 °C than 24° and 35 °C, and better in dried julienned squid than dried filefishes. At 35 °C, the populations of S. aureus were rapidly diminished and undetectable in dried filefishes and julienned squid after 14 and 19 days, respectively. SEA production did not increase during the 5-month storage period, regardless of the temperature and type of dried fish products. Although it is advised to store dried fish products at 10 °C for quality control in retail markets, refrigerated temperature is more likely to facilitate the survival of S. aureus in dried fish products. Thus, dried fish products should be produced and stored under hygienic conditions.

Protective inflammatory response against visceral leishmaniasis with potato tuber extract: A new approach of successful therapy.

The increasing number of drug resistance issue of Leishmania donovani strain to common drugs compels to develop new therapeutics against leishmaniasis with minimal toxicity. In this regard, bioactive phytocomponents may lead to the discovery of new medicines with appropriate efficiency. The important roles of Leishmania proteases in the virulence of Leishmania parasite make them very hopeful targets for the improvement of current remedial of leishmaniasis. As part of a hunt for new drugs, we have evaluated in vivo anti-leishmanial activity of serine protease inhibitor rich fraction (PTEx), isolated by sodium bisulfite extraction from potato tuber. The amastigote load of 25mg/kg body weight/day treated BALB/c mice showed 86.9% decrease in liver and 88.7% in case of spleen. This anti-leishmanial effect was also supported by PTEx induced immunomodulatory activity like acute formation of ROS and prolonged NO generation. The Th1/Th2 cytokine balance in splenocytes of PTEx treated animals was estimated and evaluated by ELISA assay as well as by mRNA expression using RT-PCR. Furthermore, significant survival rate (80%) was observed in PTEx treated hamsters. Thus, from the present observations we could accentuate the potential of PTEx to be employed as a new therapeutics from natural source against L. donovani. This might also provide a novel perception of natural serine protease inhibitor from potato tuber as an alternate approach for the treatment of visceral leishmaniasis.