Bactericidal efficacy of slightly acidic electrolyzed water (SAEW) against Listeria monocytogenes planktonic cells and biofilm on food-contact surfaces

In the present study, the bactericidal efficacy of slightly acidic electrolyzed water (SAEW) against L. monocytogenes planktonic cells and biofilm on food-contact surfaces including stainless steel and glass was systematically evaluated. The results showed that SAEW (pH of 5.09 and available chlorine concentration (ACC) of 60.33 mg/L) could kill L. monocytogenes on food-contact surfaces completely in 30 s, whose disinfection efficacy is equal to that of NaClO solutions (pH of 9.23 and ACC of 253.53 mg/L). The results showed that long exposure time and high ACC contributed to the enhancement of the disinfection efficacy of SAEW on L. monocytogenes on food-contact surfaces. Moreover, the log reduction of SAEW treatment presented an increasing tendency within the prolonging of treatment time when SAEW was used to remove the L. monocytogenes biofilm formed on stainless steel and glass surfaces, which suggested that SAEW could remove L. monocytogenes biofilm effectively and its disinfection efficacy is equal to (in case of stainless steel) or higher than (in case of glass) that of high ACC of NaClO solutions. In addition, the results of the crystal violet staining and scanning electron microscopy (SEM) also demonstrated that SAEW treatment could remove the L. monocytogenes biofilm on food-contact surfaces.

Combined in-vitro and on-farm evaluation of commercial disinfectants used against Brachyspira hyodysenteriae

Background Swine dysentery (SD) is a severe infectious disease with a relevant impact on pig production usually caused by Brachyspira hyodysenteriae, although B. hampsonii causes an identical clinical picture. SD control relies on antimicrobials, good management practices and strict biosecurity with cleaning and disinfection as crucial tools to avoid the pathogen transmission. This study evaluates the in-vitro efficacy of an array of commercial disinfectants against a collection of B. hyodysenteriae isolates using broth tests. The efficacy of cleaning and disinfection protocols was also evaluated on two farms with endemic SD using surface swabs collected in emptied pens before and after cleaning and disinfection procedures, using both real-time PCR and bacterial microbiological culture. Results Most of the commercial disinfectants evaluated were effective against all B. hyodysenteriae isolates tested, with a reduction of more than 5.00 log10 CFU/mL (bactericidal efficacy of 99.999%). However, some isolates exhibited reduced susceptibility to Virkon-S and Limoseptic disinfectants. The evaluation of cleaning and disinfection protocols on farms with SD outbreaks showed that approximately half the pens tested (n = 25) were positive by real-time PCR after pigs removal (mean B. hyodysenteriae counts 5.72 ± 1.04 log10 CFU/mL) while almost 20% of the pens remained positive after cleaning (n = 7) and disinfection (n = 5) procedures although with significantly lower, mean estimates (4.31 ± 0.43 log10 CFU/mL and 4.01 ± 0.55 log10 CFU/mL, respectively). Conclusions These results show the efficacy of disinfectants against B. hyodysenteriae but also stress the need to implement adequately the cleaning and disinfection protocols on pig farms and review and revise their efficiency periodically.

Improved bactericidal efficacy and thermostability of Staphylococcus aureus-specific bacteriophage SA3821 by repeated sodium pyrophosphate challenges

As antibiotic resistance is being a threat to public health worldwide, bacteriophages are re-highlighted as alternative antimicrobials to fight with pathogens. Various wild-type phages isolated from diverse sources have been tested, but potential mutant phages generated by genome engineering or random mutagenesis are drawing increasing attention. Here, we applied a chelating agent, sodium pyrophosphate, to the staphylococcal temperate Siphoviridae phage SA3821 to introduce random mutations. Through 30 sequential sodium pyrophosphate challenges and random selections, the suspected mutant phage SA3821M was isolated. SA3821M maintained an intact virion morphology, but exhibited better bactericidal activity against its host Staphylococcous aureus CCARM 3821 for up to 17 h and thermostability than its parent, SA3821. Sodium pyrophosphate-mediated mutations in SA3821M were absent in lysogenic development genes but concentrated (83.9%) in genes related to the phage tail, particularly in the tail tape measure protein, indicating that changes in the tail module might have been responsible for the altered traits. This intentional random mutagenesis through controlled treatments with sodium pyrophosphate could be applied to other phages as a simple but potent method to improve their traits as alternative antimicrobials.

The Bactericidal Efficacy and the Mechanism of Action of Slightly Acidic Electrolyzed Water on Listeria monocytogenes’ Survival

In the present work, the bactericidal efficacy and mechanism of slightly acidic electrolyzed water (SAEW) on L. monocytogenes were evaluated. The results showed that the strains of L. monocytogenes were killed completely within 30 s by SAEW whose available chlorine concentration (ACC) was higher than 12 mg/L, and it was confirmed that ACC is the main factor affecting the disinfection efficacy of SAEW. Moreover, our results demonstrated that SAEW could destroy the cell membrane of L. monocytogenes, which was observed by SEM and FT-IR, thus resulting in the leakage of intracellular substances including electrolyte, protein and nucleic acid, and DNA damage. On the other hand, the results found that SAEW could disrupt the intracellular ROS balance of L. monocytogenes by inhibiting the antioxidant enzyme activity, thus promoting the death of L. monocytogenes. In conclusion, the bactericidal mechanism of SAEW on L. monocytogenes was explained from two aspects including the damage of the cell membrane and the breaking of ROS balance.

SYNTHESIS OF AMIDES WITH MONOETHANOLAMINE OF AMBER AND OXALIC ACID AND RESEARCH OF THEIR BACTERICIDE PROPERTIES

Amides were synthesized in 1:1 and 1:2 molar ratios of amber and oxalic acid with monoethanolamine. IR-spectra of the obtained substances were drawn, the structures were determined and confirmed, physicochemical properties were studied. 10% aqueous solutions of these substances were prepared, physicochemical properties were determined and bactericidal properties were studied. Bactericidal efficacy of amber acid N-mono­ethanolamide was 82.9% at 25 mg/l of concentration, 88% at 50 mg/l and 95% at 100 mg/l of concentrations. Bactericidal efficacy of amber acid N1, N2 - bis-monoethanolamide was 69% at 25 mg/l of concentration, 83.2% at 50 mg/l and 90% at 100 mg/l of concentrations. The bactericidal effect of oxalic acid has also been studied N-mono­ethanolamide was 82% at 25 mg/l of concentration, 91% at 50 mg/l and 97% at 100 mg/l of concentrations. Bactericidal efficacy of amber acid N1, N2 - bis-monoethanolamide was 85% at 25 mg/l of concentration, 94% at 50 mg/l and 97.6 at 100 mg/l of concentrations. Key words: amber acid, oxalic acid, bactericidal efficacy, N-monoethanolamide, N1,N -bis-monoethanolamide