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J Appl Microbiol. 2012 Oct;113(4):824-36. doi: 10.1111/j.1365-2672.2012.05387.x. Epub 2012 Aug 2.

Effects of wet heat treatment on the germination of individual spores of Clostridium perfringens.

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  • 1Department of Physics, East Carolina University, Greenville, NC 27858-4353, USA.

Abstract

AIM:

To analyse the effect of wet heat treatment on nutrient and non-nutrient germination of individual spores of Clostridium perfringens.

METHODS AND RESULTS:

Raman spectroscopy and differential interference contrast (DIC) microscopy were used to monitor the dynamic germination of individual untreated and wet heat-treated spores of Cl. perfringens with various germinants. When incubated in water at 90-100°C for 10-30 min, more than 90% of spores were inactivated but 50-80% retained their Ca(2+) -dipicolinic acid (CaDPA). The wet heat-treated spores that lost CaDPA exhibited extensive protein denaturation as seen in the 1640-1680 cm(-1) (amide I) and 1230-1340 cm(-1) (amide III) regions of Raman spectra, while spores that retained CaDPA showed partial protein denaturation. Wet heat-treated spores that retained CaDPA germinated with KCl or l-asparagine, but wet heat treatment increased values of T(lag) , ΔT(release) and ΔT(lys) , during which spores initiated release of the majority of their CaDPA after mixing with germinant, released >90% of their CaDPA and completed the decrease in their DIC intensity because of cortex hydrolysis, respectively. Untreated Cl. perfringens spores lacking the essential cortex-lytic enzyme (CLE), SleC, exhibited longer T(lag) and ΔT(release) values during KCl germination than wild-type spores and germinated poorly with CaDPA. Wet heat-treated wild-type spores germinating with CaDPA or dodecylamine exhibited increased T(lag) , ΔT(release) and ΔT(lys) values, as did wet heat-treated sleC spores germinating with dodecylamine.

CONCLUSIONS:

(i) Some proteins important in Cl. perfringens spore germination are damaged by wet heat treatment; (ii) the CLE SleC or the serine protease CspB that activates SleC might be germination proteins damaged by wet heat; and (iii) the CaDPA release process seems likely to be damaged by wet heat.

SIGNIFICANCE AND IMPACT OF THE STUDY:

This study provides information on the germination of individual Cl. perfringens spores and improves the understanding of effects of wet heat treatment on spores.

© 2012 The Authors Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

PMID:
22776375
[PubMed - indexed for MEDLINE]
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