# Sporulation Abilities and Heat Resistance of Clostridium perfringens Strains Isolated from French Food Borne Outbreaks

## TL;DR

This study examines how different strains of Clostridium perfringens from foodborne outbreaks in France vary in their ability to form spores and resist heat, which is important for food safety.

## Contribution

The study identifies a new group of C. perfringens strains with intermediate heat resistance, expanding understanding of the species' stress adaptation.

## Key findings

- Strains in the chromosomal cpe clade showed the highest sporulation abilities and heat resistance.
- Cpe-negative strains in the chromosomal cpe clade exhibited intermediate heat resistance.
- Plasmidic cpe strains were the most heat-sensitive.

## Abstract

Clostridium perfringens is responsible for various diseases. Foodborne outbreaks (FBOs) result from the in situ production of C. perfringens enterotoxin (CPE) by type F strains during sporulation. The cpe gene can be plasmidic (p-cpe) or chromosomal (c-cpe). Strains (c-cpe) exhibit greater heat resistance and are frequently associated with FBO. Strains cpe-negative are considered heat-sensitive. This study investigates the sporulation abilities and heat resistance of eight C. perfringens strains isolated from French foodborne outbreaks. Whole-genome sequencing classified the strains into two clades: the “chromosomal cpe clade,”, mainly composed of cpe-positive strains with c-cpe and some cpe-negative strains, and the “plasmidic cpe clade,”, primarily containing cpe-negative strains and a few with plasmid-borne cpe. Sporulation assays and thermal inactivation kinetics were performed on FBO strains to evaluate the influence of genetic variability on sporulation abilities and heat resistance. Experimental analyses revealed that strains within the “chromosomal cpe clade” exhibited the highest sporulation abilities, regardless of cpe presence, while those in the “plasmidic cpe clade” had low sporulation ability. Moreover, heat-resistant spores were produced exclusively by strains of the “chromosomal cpe clade,” with c-cpe strains exhibiting the highest heat resistance (δ95 °C ≈ 49 min), followed by cpe-negative strains (δ95 °C ≈ 9.5 min). p-cpe strains exhibited a heat-sensitive phenotype, with δ85 °C values of 12 min. A key finding of this study is the identification of a group with intermediate heat resistance, distinct from the highly heat-resistant (c-cpe) and heat-sensitive (p-cpe) strains. This intermediate heat-resistance phenotype, observed in cpe-negative strains within the “chromosomal cpe clade,” offers a new perspective on C. perfringens stress adaptation, suggesting their potential for persistence in food. Their heat resistance, along with the potential for cpe gene transfer, could make these strains a relevant hazard for cooked, cooled, and re-heated meat products.