Different nutrient receptors varied in triggering germination of Bacillus subtilis spores with a pressure of 150 MPa, the GerA receptor being more responsive than the GerB receptor and even more responsive than the GerK receptor. This hierarchy in receptor responsiveness to pressure was the same as receptor responsiveness to a mixture of nutrients. The levels of nutrient receptors influenced rates of pressure germination, since the GerA receptor is more abundant than the GerB receptor and elevated levels of individual receptors increased spore germination by 150 MPa of pressure. However, GerB receptor variants with relaxed specificity for nutrient germinants responded as well as the GerA receptor to this pressure. Spores lacking dipicolinic acid did not germinate with this pressure, and pressure activation of the GerA receptor required covalent addition of diacylglycerol. However, pressure activation of the GerB and GerK receptors displayed only a partial (GerB) or no (GerK) diacylglycerylation requirement. These effects of receptor diacylglycerylation on pressure germination are similar to those on nutrient germination. Wild-type spores prepared at higher temperatures germinated more rapidly with a pressure of 150 MPa than spores prepared at lower temperatures; this was also true for spores with only one receptor, but receptor levels did not increase in spores made at higher temperatures. Changes in inner membrane unsaturated fatty acid levels, lethal treatment with oxidizing agents, or exposure to chemicals that inhibit nutrient germination had no major effect on spore germination by 150 MPa of pressure, except for strong inhibition by HgCl 2 .
Over one-half of foodborne illnesses are believed to be viral in origin. The ability of viruses to persist in the environment and foods, coupled with low infectious doses, allows even a small amount of contamination to cause serious problems. An increased incidence of foodborne illnesses and consumer demand for fresh, convenient, and safe foods have prompted research into alternative food-processing technologies. This review focuses on viral inactivation by both traditional processing technologies such as use of antimicrobial agents and the application of heat, and also novel processing technologies including high-pressure processing, ultraviolet-and gammairradiation, and pulsed electric fields. These industrially applicable control measures will be discussed in relation to the 2 most common causes of foodborne viral illnesses, hepatitis A virus and human noroviruses. Other enteric viruses, including adenoviruses, rotaviruses, aichi virus, and laboratory and industrial viral surrogates such as feline caliciviruses, murine noroviruses, bacteriophage MS2 and X174, and virus-like particles are also discussed. The basis of each technology, inactivation efficacy, proposed mechanisms of viral inactivation, factors affecting viral inactivation, and applicability to the food industry with a focus on ready-to-eat foods, produce, and shellfish, are all featured in this review.
This review focuses on the responses of microbial spores to food processes that incorporate high hydrostatic pressures. While the majority of information deals with spores of Bacillus species, spores of Clostridium and Alicyclobacillus species are also discussed, and a section of the review surveys the responses of fungal spores to high-pressure processing. The mechanisms of the germination of bacterial spores are outlined in detail with regard to spore physiology and structure, along with molecular aspects of germinants and the interaction with spore receptors. Use of treatments combining pressure and temperature for a range of different food products is reviewed, including examples of hurdle technology employing high hydrostatic pressure. Pressure-assisted thermal sterilization is also discussed.
Aims: To elucidate the factors that determine the rate of germination of Bacillus subtilis spores with very high pressure (VHP) and the mechanism of VHP germination. Methods and Results: Spores of B. subtilis were germinated rapidly with a VHP of 500 MPa at 50°C. This VHP germination did not require the spore's nutrient-germinant receptors, as found previously, and did not require diacylglycerylation of membrane proteins. However, the spore's pool of dipicolinic acid (DPA) was essential. Either of the two redundant enzymes that degrade the spore's peptidoglycan cortex, and thus allow completion of spore germination, was essential for completion of VHP germination. However, neither of these enzymes was needed for DPA release triggered by VHP treatment. Completion of spore germination as well as DPA release with VHP had an optimum temperature of approx. 60°C, in contrast to an optimum temperature of 40°C for germination with the moderately high pressure of 150 MPa. The rate of spore germination by VHP decreased approx. fourfold when the sporulation temperature increased from 23°C to 44°C, and decreased twofold when 1 mol l )1 salt was present in sporulation. However, large variations in levels of unsaturated fatty acids in the spore's inner membranes did not affect rates of VHP germination. Complete germination of spores by VHP was not inhibited significantly by killing of spores with several oxidizing agents, and was not inhibited by ethanol, octanol or o-chlorophenol at concentrations that abolish nutrient germination. Completion of spore germination by VHP was also inhibited by Hg 2+ , but this ion did not inhibit DPA release caused by VHP. In contrast, dodecylamine, a surfactant that can trigger spore germination, strongly inhibited DPA release caused by VHP treatment. Conclusions: VHP does not cause spore germination by acting upon the spore's nutrient-germinant receptors, but by directly causing DPA release. This DPA release then leads to subsequent completion of germination. VHP likely acts on the spore's inner membrane to cause DPA release, targeting either a membrane protein or the membrane itself. However, the precise identity of this target is not yet clear. Significance and Impact of the Study: There is significant interest in the use of VHP to eliminate or reduce levels of bacterial spores in foods. As at least partial spore germination by pressure is almost certainly essential for subsequent spore killing, knowledge of factors involved and the mechanism of VHP germination are crucial to the understanding of spore killing by VHP. This work provides new insight into factors that can affect the rate of B. subtilis spore germination by VHP, and into the mechanism of VHP germination itself.
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