scholarly journals Complete genome of the barotolerant Listeria monocytogenes RO15 strain and comparison with other strains isolated from food and food processing environments

2020 ◽  
Author(s):  
Ilhan Cem Duru ◽  
Margarita Andreevskaya ◽  
Pia Laine ◽  
Tone Mari Rode ◽  
Anne Ylinen ◽  
...  

Abstract Background Consumption of Listeria monocytogenes contaminated food can cause infection with a high mortality rate in humans and animals. High pressure processing (HPP) is a non-thermal preservation technique adopted by the food industry to inactivate food pathogens, including L. monocytogenes. Strains of L. monocytogenes show different level of resistance to the high pressure. Some strains resist up to 500 MPa pressure. Here, we tested the pressure resistance of 10 different L. monocytogenes strains, including field isolates and widely used type strains, to 400 and 600 MPa pressure treatments. Genome sequencing, and genome comparison of the tested L. monocytogenes strains were performed to investigate the relation between genomic profile and pressure resistance. Results In this study, we showed that none of the tested strains were resistant to 600 MPa, more than 5 log 10 reduction observed for all strains after 1 minute 600 MPa pressure treatment. However, L. monocytogenes strain RO15 showed no significant reduction in viable cell counts after 400 MPa for 1 minute and it was defined as barotolerant. Genome sequencing of so far unsequenced L. monocytogenes strain RO15, 2HF33, MB5, AB199, AB120, C7, and RO4 allowed us to compare their gene content. Genome comparison of 10 tested strains showed that the three most pressure tolerant strains had more than one CRISPR system with self-targeting spacers. Further, several anti-CRISPR genes were detected in these strains. Pan-genome wide analysis showed that 10 prophage genes were significantly associated with the three most barotolerant strains. Conclusions L. monocytogenes strain RO15 was the most pressure tolerant among the selected strains. Genome comparison suggests that there might be a relationship with prophages, CRISPR systems and pressure resistance in L. monocytogenes .

2020 ◽  
Author(s):  
Ilhan Cem Duru ◽  
Margarita Andreevskaya ◽  
Pia Laine ◽  
Tone Mari Rode ◽  
Anne Ylinen ◽  
...  

Abstract Background: High pressure processing (HPP; i.e. 100 - 600 MPa pressure depending on product) is a non-thermal preservation technique adopted by the food industry to decrease significantly foodborne pathogens, including Listeria monocytogenes, from food. However, susceptibility towards pressure differs among diverse strains of L. monocytogenes and it is unclear if this is related to their genomic content. Here, we tested the barotolerance of 10 different L. monocytogenes strains, from food and food processing environments and widely used reference type strains, to pressure treatments with 400 and 600 MPa. Genome sequencing and genome comparison of the tested L. monocytogenes strains were performed to investigate the relation between genomic profile and pressure tolerance.Results: None of the tested strains were tolerant to 600 MPa. A reduction of more than 5 log10 was observed for all strains after 1 minute 600 MPa pressure treatment. L. monocytogenes strain RO15 showed no significant reduction in viable cell counts after 400 MPa for 1 minute and was therefore defined as barotolerant. Genome analysis of so far unsequenced L. monocytogenes strain RO15, 2HF33, MB5, AB199, AB120, C7, and RO4 allowed us to compare the gene content of all strains tested. This revealed that the three most pressure tolerant strains had more than one CRISPR system with self-targeting spacers. Furthermore, several anti-CRISPR genes were detected in these strains. Pan-genome analysis showed that 10 prophage genes were significantly associated with the three most barotolerant strains.Conclusions: L. monocytogenes strain RO15 was the most pressure tolerant among the selected strains. Genome comparison suggests that there might be a relationship between prophages and pressure tolerance in L. monocytogenes.


2003 ◽  
Vol 66 (11) ◽  
pp. 2057-2061 ◽  
Author(s):  
ABDULLATIF TAY ◽  
THOMAS H. SHELLHAMMER ◽  
AHMED E. YOUSEF ◽  
GRADY W. CHISM

The objectives of this study were to investigate the variability among Listeria monocytogenes strains in response to high-pressure processing, identify the most resistant strain as a potential target of pressure processing, and compare the inactivation kinetics of pressure-resistant and pressure-sensitive strains under a wide range (350 to 800 MPa) of pressure treatments. The pressure resistance of Listeria innocua and nine strains of L. monocytogenes was compared at 400 or 500 MPa and 30°C. Significant variability among strains was observed. The decrease in log CFU/ml during the pressure treatment was from 1.4 to 4.3 at 400 MPa and from 3.9 to >8 at 500 MPa. L. monocytogenes OSY-8578 exhibited the greatest pressure resistance, Scott A showed the greatest pressure sensitivity, and L. innocua had intermediate resistance. On the basis of these findings, L. monocytogenes OSY-8578 is a potential target strain for high-pressure processing efficacy studies. The death kinetics of L. monocytogenes Scott A and OSY-8578 were investigated at 350 and 800 MPa. Survivors at 350 MPa were enumerated by direct plating, and survivors at 800 MPa were enumerated by the most-probable-number technique. Both pressure-resistant and pressure-sensitive strains exhibited non–first-order death behavior, and excessive pressure treatment did not eliminate the tailing phenomenon.


Biomolecules ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 677
Author(s):  
Cristina Serra-Castelló ◽  
Ilario Ferrocino ◽  
Anna Jofré ◽  
Luca Cocolin ◽  
Sara Bover-Cid ◽  
...  

Formulations with lactate as an antimicrobial and high-pressure processing (HPP) as a lethal treatment are combined strategies used to control L. monocytogenes in cooked meat products. Previous studies have shown that when HPP is applied in products with lactate, the inactivation of L. monocytogenes is lower than that without lactate. The purpose of the present work was to identify the molecular mechanisms underlying the piezo-protection effect of lactate. Two L. monocytogenes strains (CTC1034 and EGDe) were independently inoculated in a cooked ham model medium without and with 2.8% potassium lactate. Samples were pressurized at 400 MPa for 10 min at 10 °C. Samples were subjected to RNA extraction, and a shotgun transcriptome sequencing was performed. The short exposure of L. monocytogenes cells to lactate through its inoculation in a cooked ham model with lactate 1h before HPP promoted a shift in the pathogen’s central metabolism, favoring the metabolism of propanediol and ethanolamine together with the synthesis of the B12 cofactor. Moreover, the results suggest an activated methyl cycle that would promote modifications in membrane properties resulting in an enhanced resistance of the pathogen to HPP. This study provides insights on the mechanisms developed by L. monocytogenes in response to lactate and/or HPP and sheds light on the understanding of the piezo-protective effect of lactate.


2012 ◽  
Vol 75 (3) ◽  
pp. 497-503 ◽  
Author(s):  
YANG LIU ◽  
MIRKO BETTI ◽  
MICHAEL G. GÄNZLE

This study evaluated the high pressure inactivation of Campylobacter jejuni, Escherichia coli, and poultry meat spoilage organisms. All treatments were performed in aseptically prepared minced poultry meat. Treatment of 19 strains of C. jejuni at 300 MPa and 30°C revealed a large variation of pressure resistance. The recovery of pressure-induced sublethally injured C. jejuni depended on the availability of iron. The addition of iron content to enumeration media was required for resuscitation of sublethally injured cells. Survival of C. jejuni during storage of refrigerated poultry meat was analyzed in fresh and pressure-treated poultry meat, and in the presence or absence of spoilage microbiota. The presence of spoilage microbiota did not significantly influence the survival of C. jejuni. Pressure treatment at 400 MPa and 40°C reduced cell counts of Brochothrix thermosphacta, Carnobacterium divergens, C. jejuni, and Pseudomonas fluorescens to levels below the detection limit. Cell counts of E. coli AW1.7, however, were reduced by only 3.5 log (CFU/g) and remained stable during subsequent refrigerated storage. The resistance to treatment at 600 MPa and 40°Cof E. coli AW1.7 was compared with Salmonella enterica, Shiga toxin–producing E. coli and nonpathogenic E. coli strains, and Staphylococcus spp. Cell counts of all organisms except E. coli AW 1.7 were reduced by more than 6 log CFU/g. Cell counts of E. coli AW1.7 were reduced by 4.5 log CFU/g only. Moreover, the ability of E. coli AW1.7 to resist pressure was comparable to the pressure-resistant mutant E. coli LMM1030. Our results indicate that preservation of fresh meat requires a combination of high pressure with high temperature (40 to 60°C) or other antimicrobial hurdles.


2017 ◽  
Vol 84 (2) ◽  
Author(s):  
Sandeep Tamber

ABSTRACTHigh-pressure processing is a nonthermal method of food preservation that uses pressure to inactivate microorganisms. To ensure the effective validation of process parameters, it is important that the design of challenge protocols consider the potential for resistance in a particular species. Herein, the responses of 99 diverseSalmonella entericastrains to high pressure are reported. Members of this population belonged to 24 serovars and were isolated from various Canadian sources over a period of 26 years. When cells were exposed to 600 MPa for 3 min, the average reduction in cell numbers for this population was 5.6 log10CFU/ml, with a range of 0.9 log10CFU/ml to 6 log10CFU/ml. Eleven strains, from 5 serovars, with variable levels of pressure resistance were selected for further study. The membrane characteristics (propidium iodide uptake during and after pressure treatment, sensitivity to membrane-active agents, and membrane fatty acid composition) and responses to stressors (heat, nutrient deprivation, desiccation, and acid) for this panel suggested potential roles for the cell membrane and the RpoS regulon in mediating pressure resistance inS. enterica. The data indicate heterogeneous and multifactorial responses to high pressure that cannot be predicted for individualS. entericastrains.IMPORTANCEThe responses of foodborne pathogens to increasingly popular minimal food decontamination methods are not understood and therefore are difficult to predict. This report shows that the responses ofSalmonella entericastrains to high-pressure processing are diverse. The magnitude of inactivation does not depend on how closely related the strains are or where they were isolated. Moreover, strains that are resistant to high pressure do not behave similarly to other stresses, suggesting that more than one mechanism might be responsible for resistance to high pressure and the mechanisms used may vary from one strain to another.


2009 ◽  
Vol 72 (1) ◽  
pp. 165-168 ◽  
Author(s):  
JOSEPH E. SCHLESSER ◽  
BRIAN PARISI

In 2003, the U.S. Department of Health and Human Services announced a new research program to develop technologies and strategies to prevent and minimize potential food safety and security threats. The threat of terrorist attacks against the nation's food supplies has created the need to study microorganisms not typically associated with foodborne illness. High-pressure processing has been proposed as a treatment to reduce Yersinia pestis and Francisella tularensis LVS levels in beverages. The objectives of this work were to determine the pressure resistance of Y. pseudotuberculosis 197 (surrogate for Y. pestis) and F. tularensis LVS (vaccine strain). For each bacterium, samples of ultrahigh-temperature pasteurized skim milk and pasteurized reduced-acid orange juice (pH ca. 4.2) were inoculated at a minimum level of 5 log CFU/ml. Ten-milliliter samples of the inoculated product were vacuum sealed in polyester pouches and subjected to pressures of 300 and 500 MPa for holding times ranging from 30 s to 6 min. One set of trials was performed at an initial temperature of 10°C and another at 25°C. Processed samples were immediately plated and enumerated. A pressure treatment of 300 MPa at 25°C for less than 6 min was not sufficient to achieve a 5-log reduction of Y. pseudotuberculosis 197 or F. tularensis LVS in milk. However, a pressure treatment of 500 MPa was effective at hold times as low as 30 s. Overall, F. tularensis LVS demonstrated less pressure resistance than Y. pseudotuberculosis 197. Based on these findings, a high-pressure process designed to inactivate 5 log CFU of Y. pseudotuberculosis 197 per ml and F. tularensis LVS in orange juice or milk should be set at or above 500 MPa with a hold time of 2 min or greater.


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