Impact of temperature application and concentration of commercial sanitizers on inactivation of food-plant fungal spores.

This study aimed at quantifying the impact of the concentration of four commercial sanitizers and temperature on mold spores inactivation. The sanitizers were based on the following fungicide molecules, ethanol (ARVO 21 SR), active chlorine (ARVO CLM 600), hydrogen peroxide (Nocolyse Food) and triamine (P3 Topax 960). Food plant spores were produced under a moderate water stress, 0.95 aw and dry-harvested to simulate airborne spores responsible for contamination in the food industry. First, Aspergillus flavus, Cladosporium cladosporioides, Mucor circinelloides, and two Penicillium commune isolates were tested against the sanitizers at 20 °C and at a concentration recommended by the manufacturers. Overall, A. flavus was the less resistant species. Second the effects of concentration and temperature were assessed on the most resistant species, i.e., P. commune UBOCC-A-116003 (ARVO 21 SR and P3 Topax 960), P. commune UBOCC-A-112059 (ARVO CLM 600), and M. circinelloides (Nocolyse Food). With the exception of ARVO 21 SR, the observed inactivation kinetics were downward concave. The time necessary to obtain 4 log reduction, t4D, was estimated by means of the Weibull model. At 20 °C and at the recommended concentration by the manufacturers, t4D (min) for the most resistant strains were equal to 2.14 (ARVO 21 SR), 7.35 (ARVO CLM 600), 39.3 (Nocolyse Food) and 82.8 (P3 Topax 960). T4D was increased at lower concentrations and temperatures. These effects were more pronounced for ARVO 21 SR, t4D were about 10 fold and 20 fold the above reported value, 2.14 min, at 8 °C and by diluting the sanitizer by a 10:8 factor, respectively. The least effect of temperature, 3 fold, was shown for ARVO CLM 600, while concentration of P3 Topax 960 had no significant effect on t4D within the recommended utilization range.

Impact of the physiological state of fungal spores on their inactivation by active chlorine and hydrogen peroxide.

This study aimed at assessing the impact of the physiological state of fungal spores on inactivation by sodium hypochlorite, 0.1% and 0.2% active chlorine, and 3% hydrogen peroxide. In this context, two physiological states were compared for 4 fungal species (5 strains). The first physiological state corresponded to fungal spores produced at 0.99 aw and harvested using an aqueous solution (laboratory conditions), while the second one corresponded to fungal spores produced under a moderate water stress (0.95 aw) and dry-harvested (mechanical harvesting without use of any water, mimicking food plant conditions). Aspergillus flavus "food plant" conidia were more resistant to all tested fungicide molecules than the "laboratory" ones. The same phenomenon was observed for Penicillium commune UBOCC-A-116003 conidia treated with hydrogen peroxide. However, this isolate did not exhibit any inactivation difference between "laboratory" and "food plant" conidia treated with sodium hypochlorite. Similarly, the physiological state of Cladosporium cladosporioides conidia did not impact the efficacy of the tested biocides. P. commune UBOCC-A-112059 "food plant" and "laboratory" conidia were more resistant to hydrogen peroxide and sodium hypochlorite, respectively. As for Mucor circinelloides, "laboratory" spores were more resistant to all disinfectant than the "food plant" ones. Noteworthy, regardless of the physiological state, all M. circinelloides and C. cladosporioides conidia were inactivated for 5 min treatment at 0.2% active chlorine and for 2.5 min treatment at 0.1% active chlorine, while the conidia of all the other species remained viable for these treatments. The obtained data indicate that the efficacy of disinfectant molecules depends not only on the encountered fungal species and its intraspecific diversity but also on the spore physiological state.

Impact of intraspecific variability and physiological state on Penicillium commune inactivation by 70% ethanol.

The aim of this study was to assess the impact of the physiological state and intraspecific variability on the efficacy of 70% ethanol to inactivate conidia of Penicillium commune, used as a representative species of dairy product contaminants. Four physiological states were obtained by modifying the water activity during the production of conidia (0.995 and 0.950) and the harvesting conditions (hydrated and non-hydrated). These conditions were applied to four different P. commune strains isolated from contaminated dairy products. Five minutes exposure to 70% ethanol at ambient temperature allowed total inactivation of conidia (>4 log10) regardless of the physiological state or the strain. For 1 min exposure, regardless of the strains, only dry-harvested conidia produced at aw 0.950 exhibited survivors. Survival after 2 min exposure was observed for this physiological state for P. commune UBOCC-A-116003 only. For this strain, the impact of the physiological state was greater than 1.54 log10 between dry-harvested conidia produced at aw 0.950 that exhibited survivors after 1 min treatment and the 3 other kinds of conidia that were all inactivated. For 1 min exposure, by comparing the more resistant strain to the three other strains, the impact of the intraspecific variability was 2.35 log10. These results demonstrated that the physiological state of the conidia, the representativeness of the tested species and strains should be taken into account to assess the efficacy of disinfectants in dairies.

Modulation of antiviral immunity by the ichnovirus HdIV in Spodoptera frugiperda.

Polydnaviruses (PDVs) are obligatory symbionts found in thousands of endoparasitoid species and essential for successful parasitism. The two genera of PDVs, ichnovirus (IV) and bracovirus (BV), use different sets of virulence factors to ensure successful parasitization of the host. Previous studies have shown that PDVs target apoptosis, one of the innate antiviral responses in many host organisms. However, IV and BV have been shown to have opposite effects on this process. BV induces apoptosis in host cells, whereas some IV proteins have been shown to have anti-apoptotic activity. The different biological contexts in which the assays were performed may account for this difference. In this study, we evaluated the interplay between apoptosis and the ichnovirus HdIV from the parasitoid Hyposoter didymator, in the HdIV-infected hemocytes and fat bodies of S. frugiperda larvae, and in the Sf9 insect cell line challenged with HdIV. We found that HdIV induced cell death in hemocytes and fat bodies, whereas anti-apoptotic activity was observed in HdIV-infected Sf9 cells, with and without stimulation with viral PAMPs or chemical inducers. We also used an RT-qPCR approach to determine the expression profiles of a set of genes known to encode key components of the other main antiviral immune pathways described in insects. The analysis of immune gene transcription highlighted differences in antiviral responses to HdIV as a function of host cell type. However, all these antiviral pathways appeared to be neutralized by low levels of expression for the genes encoding the key components of these pathways, in all biological contexts. Finally, we investigated the effect of HdIV on the general antiviral defenses of the lepidopteran larvae in more detail, by studying the survival of S. frugiperda co-infected with HdIV and the entomopathogenic densovirus JcDV. Coinfected S. frugiperda larvae have increased resistance to JcDV at an early phase of infection, whereas HdIV effects enhance the virulence of the virus at later stages of infection. Overall, these results reveal complex interactions between HdIV and its cellular environment.