Inactivation of Clostridium botulinum nonproteolytic type B spores by high pressure processing at moderate to elevated high temperatures

Reddy, N. R.; Tetzloff, R.C.; Solomon, Haim M.; Larkin, John W.

doi:10.1016/j.ifset.2006.03.002

Cited by 72 publications

(39 citation statements)

References 23 publications

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“…The effect of the intrinsic properties of food on spore inactivation by HPT processing has been explored by several research groups (2,18,21,28,29,30). Criteria for choosing the model products for this work were based partially on their various intrinsic properties, namely, pH and fat and starch contents; although these factors were not assessed systematically, it is worth considering the effects of the various products on spore inactivation, since no published inactivation studies have been performed with similar products.…”

Section: Discussionmentioning

confidence: 99%

“…For mashed carrot, a 12-min process at 600 MPa and 80°C was required to inactivate a proteolytic type A C. botulinum strain by 5 log 10 , whereas a proteolytic type B C. botulinum strain was inactivated by Ͻ3 log 10 by a process of 600 MPa at 80°C for 60 min (18). Meat and carrot broths inoculated with 5 log 10 spores/ml Clostridium sporogenes PA 3679, the nonpathogenic surrogate for C. botulinum for thermal processing studies, were sterilized only with pressure treatments of Ͼ800 MPa for 5 min at initial temperatures of 80 to 90°C (16).Most studies comparing the heat-only and high-pressureplus-heat resistances of bacterial spores have concluded that, in most cases, pressure and heat do act synergistically to deliver lethality (1,15,17,27,28,30,33). Predominantly, the approaches used by others to demonstrate synergy assume loglinear inactivation kinetics during the pressure hold phase of the high-pressure thermal (HPT) process, ignore inactivation during the pressure come-up time (CUT) and decompression, derive decimal reduction times (D T values; the time required at a constant temperature [T] to achieve a decimal reduction in the number of surviving spores) under constant pressure conditions, and compare these with D T values determined at T and ambient pressure.…”

mentioning

confidence: 99%

“…Most studies comparing the heat-only and high-pressureplus-heat resistances of bacterial spores have concluded that, in most cases, pressure and heat do act synergistically to deliver lethality (1,15,17,27,28,30,33). Predominantly, the approaches used by others to demonstrate synergy assume loglinear inactivation kinetics during the pressure hold phase of the high-pressure thermal (HPT) process, ignore inactivation during the pressure come-up time (CUT) and decompression, derive decimal reduction times (D T values; the time required at a constant temperature [T] to achieve a decimal reduction in the number of surviving spores) under constant pressure conditions, and compare these with D T values determined at T and ambient pressure.…”

mentioning

confidence: 99%

“…Predominantly, the approaches used by others to demonstrate synergy assume loglinear inactivation kinetics during the pressure hold phase of the high-pressure thermal (HPT) process, ignore inactivation during the pressure come-up time (CUT) and decompression, derive decimal reduction times (D T values; the time required at a constant temperature [T] to achieve a decimal reduction in the number of surviving spores) under constant pressure conditions, and compare these with D T values determined at T and ambient pressure. In such a manner, the combination of high pressure and heat has been determined to be more effective than heat only for inactivation of C. sporogenes PA3679 (1,15,33), C. botulinum 17B (proteolytic type B) (30), Clostridium tyrobutyricum ATCC 25755 (1), Thermoanaerobacterium thermosaccharolyticum ATCC 27384 (1), Geobacillus stearothermophilus (1,15,26,28), and Bacillus amyloliquefaciens TMW 2.479, TMW 2.482, and ATCC 49763 (1,27).…”

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confidence: 99%

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Synergistic Inactivation of Spores of ProteolyticClostridium botulinumStrains by High Pressure and Heat Is Strain and Product Dependent

Bull

Olivier

Diepenbeek³

et al. 2009

Appl Environ Microbiol

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The combined high pressure and heat resistances of spores of five proteolytic Clostridium botulinum strains and of the nonpathogenic surrogate strain Clostridium sporogenes PA3679 were compared with their heat-only resistances on the basis of equivalent accumulated thermal lethality, expressed as equivalent minutes at a reference temperature of 105°C (F 105°C ). Comparisons were made with three model (i.e., diluted) products, namely, 30% (wt/wt) Bolognese sauce, 50% (wt/wt) cream sauce, and rice water agar. Pressure was determined to act synergistically with heat during high-pressure thermal (HPT) processing for C. botulinum FRRB 2802 (NCTC 7273) and C. botulinum FRRB 2804 (NCTC 3805 and 62A) in the Bolognese and cream sauces and for C. botulinum FRRB 2807 (213B) in the Bolognese sauce only. No synergy was observed for C. botulinum FRRB 2803 (NCTC 2916) or FRRB 2806 (62A) or C. sporogenes FRRB 2790 (NCTC 8594 and PA3679) in any of the model products. No significant protective effect of pressure against spore inactivation was determined for any Clostridium strain in any product. Because synergy was not consistently observed among strains of C. botulinum or among products, the prediction of inactivation of C. botulinum spores by HPT sterilization (HPTS) for the present must assume a complete lack of synergy. Therefore, any HPTS process for low-acid shelf-stable foods must be at least thermally equivalent to an F 0 process of 2.8 min, in line with current good manufacturing practices. The results of this study suggest that the use of C. sporogenes PA3679 as a surrogate organism may risk overestimating inactivation of C. botulinum by HPT processing.Low-acid shelf-stable products that are microbiologically safe and stable are not obtainable by high-pressure processing (HPP) at near-ambient temperatures, as bacterial spores can survive pressures above 1,500 MPa (3,6,16,35), which far exceed the pressure capabilities of current commercial HPP equipment. Extensive inactivation of bacterial spores by high pressure is likely only to be realized in combination with initial process temperatures that exceed 60°C (16,20,23,24,31,37,39; C. M. Roberts and D. G. Hoover, presented at the Institute of Food Technologists 1996 annual meeting). Of particular interest (and concern) for low-acid shelf-stable foods is the ability of a combined high pressure and heat process to synergistically inactivate spores of the major bacterial spore-forming pathogens of concern, which are proteolytic strains of the neurotoxigenic species Clostridium botulinum.The combined high pressure and heat resistances of spores of a number of proteolytic C. botulinum strains in a number of different matrices have been investigated. Proteolytic C. botulinum type A BS-A and 62A spores were inactivated by 2 and 3 log 10 , respectively, in phosphate buffer (0.067 M, pH 7.0) after 20 min at 827 MPa at 75°C and by 3.2 log 10 and 2.7 log 10 , respectively, in a crabmeat blend (pH 7.2 to 7.4) after 15 min (29). For mashed carrot, a 12-min process at 600 MPa and 80°C ...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Synergistic Inactivation of Spores of ProteolyticClostridium botulinumStrains by High Pressure and Heat Is Strain and Product Dependent

Bull

Olivier

Diepenbeek³

et al. 2009

Appl Environ Microbiol

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“…However, the increasing consumer demand for fresh-tasting, healthy ready-to-eat foods that have been minimally processed and contain less salt and no artificial preservatives yet have a long shelf-life represents a challenge for the food industry in view of these botulinum safety guidelines (10,11,12). Furthermore, novel food processing and preservation technologies (e.g., high-pressure or pulsed electric field treatment and natural preservatives) have found their way to commercial food production, but data regarding their efficiency in controlling gIICb is scarce or lacking (13,14,15,16,17). This is in sharp contrast to the attention given in this context to other pathogens, like Listeria monocytogenes, Salmonella, and enterohemorrhagic Escherichia coli.…”

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Construction of Nontoxigenic Mutants of Nonproteolytic Clostridium botulinum NCTC 11219 by Insertional Mutagenesis and Gene Replacement

Clauwers

Vanoirbeek

Delbrassinne

et al. 2016

Appl Environ Microbiol

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Group II nonproteolytic Clostridium botulinum (gIICb) strains are an important concern for the safety of minimally processed ready-to-eat foods, because they can grow and produce botulinum neurotoxin during refrigerated storage. The principles of control of gIICb by conventional food processing and preservation methods have been well investigated and translated into guidelines for the food industry; in contrast, the effectiveness of emerging processing and preservation techniques has been poorly documented. The reason is that experimental studies with C. botulinum are cumbersome because of biosafety and biosecurity concerns. In the present work, we report the construction of two nontoxigenic derivatives of the type E gIICb strain NCTC 11219. In the first strain, the botulinum toxin gene (bont/E) was insertionally inactivated with a retargeted intron using the ClosTron system. In the second strain, bont/E was exchanged for an erythromycin resistance gene using a new gene replacement strategy that makes use of pyrE as a bidirectional selection marker. Growth under optimal and stressed conditions, sporulation efficiency, and spore heat resistance of the mutants were unaltered, except for small differences in spore heat resistance at 70°C and in growth at 2.3% NaCl. The mutants described in this work provide a safe alternative for basic research as well as for food challenge and process validation studies with gIICb. In addition, this work expands the clostridial genetic toolbox with a new gene replacement method that can be applied to replace any gene in gIICb and other clostridia. IMPORTANCEThe nontoxigenic mutants described in this work provide a safe alternative for basic research as well as for food challenge and process validation studies with psychrotrophic Clostridium botulinum. In addition, this work expands the clostridial genetic toolbox with a new gene replacement method that can be applied to replace any gene in clostridia. Botulism, caused by the botulinum neurotoxin (BoNT) produced by Clostridium botulinum, is a rare but severe paralytic illness in humans and animals. Botulinum toxins are 150-kDa proteins with zinc endopeptidase activity, consisting of two subunits, a 100-kDa heavy chain and a 50-kDa light chain. The heavy chain is responsible for binding and translocation of the light chain into the cytosol of neuronal cells, whereas the light chain cleaves SNARE proteins that are involved in the docking of acetylcholine-containing vesicles and fusion to the presynaptic membrane. When the SNARE proteins are cleaved, neurotransmitter release is inhibited, leading to the paralysis of the corresponding muscle (1, 2).C. botulinum is a strictly anaerobic bacterium that thrives in decaying organic matter in soils and sediments of ponds, lakes, and oceans. It also forms dormant endospores that are highly resilient to hostile conditions and therefore can be found widespread in the environment. These spores may contaminate foods via raw materials or other sources, possibly leading to food-borne botuli...

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Pressure and Heat Resistance of Clostridium Botulinum and Other Endospores

Gänzle¹,

Margosch²,

Buckow³

et al. 2007

High Pressure Processing of Foods

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Inactivation of Clostridium botulinum nonproteolytic type B spores by high pressure processing at moderate to elevated high temperatures

Cited by 72 publications

References 23 publications

Synergistic Inactivation of Spores of ProteolyticClostridium botulinumStrains by High Pressure and Heat Is Strain and Product Dependent

Synergistic Inactivation of Spores of ProteolyticClostridium botulinumStrains by High Pressure and Heat Is Strain and Product Dependent

Construction of Nontoxigenic Mutants of Nonproteolytic Clostridium botulinum NCTC 11219 by Insertional Mutagenesis and Gene Replacement

Pressure and Heat Resistance of Clostridium Botulinum and Other Endospores

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