Survival of Clostridioides difficile in finished dairy compost under controlled conditions.

AIM: The survival of Clostridioides difficile (previously Clostridium difficile) vegetative cells and endospores was compared at different levels of indigenous microflora using autoclaved and unautoclaved dairy composts with different moisture contents (MCs). METHODS AND RESULTS: Both types of composts adjusted to 20, 30 and 40% MCs were inoculated with a suspension of C. difficile that contained both vegetative cells (c. 5-6 log CFU per gram) and endospores (c. 5·0 CFU per gram), and then stored aerobically inside a humidity-controlled chamber at room temperature 22·5 ± 0·8°C for 1 year. The level of indigenous microflora was very stable during the storage after day 7 in both types of compost. The greatest reductions of C. difficile vegetative cell counts occurred during the first 24 h of storage in autoclaved and unautoclaved composts, which had 4·7 and 5·5 log CFU per gram with 20% MC, 1·8 and 2·1 log CFU per gram with 30% MC, and 2·3 and 1·3 log CFU per gram with 40% MC, respectively. Both MC and the duration of storage have significant (P < 0·05) effects on the survival of vegetative cells for first 120 days of storage. The slow inactivation of C. difficile vegetative cells at higher MCs during aerobic storage was confirmed by exponentially decaying modelling data during the early stage of aerobic exposure. The reduction of endospore counts (<1·0 log CFU per gram) during the storage for both types of compost at all MCs was not significant (P > 0·05) except for the autoclaved compost with 30% MC. CONCLUSION: The highly resistant C. difficile endospores to the unfavourable environmental conditions survived for more than a year while vegetative cells died off exponentially upon the initial aerobic exposure. SIGNIFICANCE AND IMPACT OF THE STUDY: The long-term survival of C. difficile endospores in contaminated compost may transmit the pathogen to fresh produce, animals or water in pre-harvest conditions.

Efficacy of novel aqueous photo-chlorine dioxide against a human norovirus surrogate, bacteriophage MS2 and Clostridium difficile endospores, in suspension, on stainless steel and under greenhouse conditions.

AIMS: The efficacy of a novel photochemical method for generating chlorine dioxide (photoClO2 ) was evaluated against human noroviruses (HuNoV) surrogate, bacteriophage MS2, and Clostridium difficile endospores. METHODS AND RESULTS: Chlorine dioxide was generated by mixing 1% sodium chlorite with 10 parts-per-million (ppm) Eosin Y and irradiating with a photo-activator-excitable light. PhotoClO2 efficacy was assessed against bacteriophage MS2 and C. difficile endospores in suspension, on hard surfaces and greenhouse conditions under soiled and unsoiled conditions. The estimated effective photoClO2 produced and consumed was 20·39 ± 0·16 ppm at a rate of 8·16 ppm per min in a 1% sodium chlorite solution. In suspension, MS2 phage was reduced by 3·35 and >5·10 log10 PFU per ml in 120 and 90 min, with and without soil, respectively. At the same time, when dried on stainless steel surface, MS2 phage was reduced by >4·53 log10 PFU per carrier in 30 min under both conditions. On the other hand, C. difficile endospores in suspension were reduced by 2·26 and 3·65 log10 CFU per ml in 120 min with and without soiling, respectively. However, on stainless steel surface, maximal reductions of the C. difficile endospores were 0·8 and 1·5 log10 CFU per carrier with and without soiling, respectively, and a maximal reduction of 2·97 log10 CFU per carrier under greenhouse conditions at 24 h. CONCLUSIONS: Overall, photoClO2 showed promise as a technology to control HuNoV contamination on environmental surfaces but requires further optimization and testing against C. difficile endospores. SIGNIFICANCE AND IMPACT OF THE STUDY: Results from this investigation will serve as a model for how to generate and quantify photoClO2 and how to appropriately evaluate this new class of disinfectants against environmentally resilient pathogens: viruses and bacterial endospores.

Thermal resistance of Clostridium difficile endospores in dairy compost upon exposure to wet and dry heat treatments.

AIM: Thermal resistance of Clostridium difficile endospores in finished dairy compost was compared at 55 and 65°C under wet and dry heat conditions. METHODS AND RESULTS: A three-strain cocktail of C. difficile endospores was inoculated into dairy compost to a final concentration of c. 5·5 log CFU per gram and the moisture content (MC) of the compost was adjusted to be 20, 30 and 40%. For the dry heat treatment at 55 and 65°C, the compost samples were placed in an environmental chamber, whereas for the wet heat treatment, the inoculated compost samples were placed in a tray submerged in a water bath. The MCs of composts were maintained well throughout the wet heat treatment while the dry heat treatment reduced the MCs of composts to <10% by the end of come-up time. During the come-up time, the log endospore reductions at a selected temperature were not significantly different in compost with three selected MCs, in each heat treatment. During the holding time, endospore counts reduced by <0·5 log CFU per gram at 55 and 65°C of dry heat treatment, whereas 0·7-0·8 and 0·6-3·0 log CFU per gram reductions were observed at 55 and 65°C in wet heat treatment respectively. CONCLUSION: The recommended minimum composting guidelines were not sufficient to reduce C. difficile endospore counts to an undetectable level (five endospores per gram). Increasing the temperature of thermophilic phase to 65°C, and maintaining higher MCs of composting feedstocks have significant (P < 0·05) effects on the endospore inactivation. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study identified factors that significantly affecting the thermal resistance of C. difficile endospores during composting, and the results suggest the current composting guidelines need to be amended in order to reduce the dissemination of C. difficile endospores in agricultural environment.