sodium-hypochlorite and Dehydration

Dry hospital environments are contaminated with pathogenic bacteria in biofilms, which suggests that current cleaning practices and disinfectants are failing.. To test the efficacy of sodium hypochlorite solution against Staphylococcus aureus dry-surface biofilms.. The Centers for Disease Control and Prevention Biofilm Reactor was adapted to create a dry-surface biofilm, containing 1.36 × 10(7)S. aureus/coupon, by alternating cycles of growth and dehydration over 12 days. Biofilm was detected qualitatively using live/dead stain confocal laser scanning microscopy (CLSM), and quantitatively with sonicated viable plate counts and crystal violet assay. Sodium hypochlorite (1000-20,000parts per million) was applied to the dry-surface biofilm for 10min, coupons were rinsed three times, and residual biofilm viability was determined by CLSM, plate counts and prolonged culture up to 16 days. Isolates before and after exposure underwent minimum inhibitory concentration (MIC) and minimum eradication concentration (MEC) testing, and one pair underwent whole-genome sequencing.. Hypochlorite exposure reduced plate counts by a factor of 7 log10, and reduced biofilm biomass by a factor of 100; however, staining of residual biofilm showed that live S. aureus cells remained. On prolonged incubation, S. aureus regrew and formed biofilms. Post-exposure S. aureus isolates had MICs and MECs that were not significantly different from the parent strains. Whole-genome sequencing of one pre- and post-exposure pair found that they were virtually identical.. Hypochlorite exposure led to a 7-log kill but the organisms regrew. No resistance mutations occurred, implying that hypochlorite resistance is an intrinsic property of S. aureus biofilms. The clinical significance of this warrants further study.

Topics: Biofilms; Colony Count, Microbial; Dehydration; Disinfectants; Environmental Microbiology; Infection Control; Microbial Sensitivity Tests; Microbial Viability; Microscopy, Confocal; Sodium Hypochlorite; Staphylococcus aureus; Surface Properties

There is an increasing need for methods of cryopreservation of arthropods. In particular, Lepidoptera are extremely important in entomological applications for the protection of agricultural crops and forest ecosystems and also in many aspects of biodiversity conservation. Yet, few studies have dealt with cryopreservation techniques in species of this insect order. The aim of this study was to examine the chill sensitivity of eggs of the greater wax moth Galleria mellonella (L.) and the possibility to cryopreserve the eggs by vitrification methods. One day-old eggs were dechlorinated with water solutions of 1.25% sodium hypochlorite and 0.04% Tween 80, treated with cryoprotective agents in two steps, subjected to rapid cooling by immersion in LN and stored in a mechanical freezer for 48 h at -140 degrees C. They exhibited survival rates of 1.6+/-0.5% after being cooled in LN and 0.6+/-0.2% after being stored in the mechanical freezer. 92.9% of the larvae that hatched from cryopreserved eggs completed development regularly, producing adults that bred and laid fertile eggs. The hatching rate of eggs in the F1 and F2 generations was higher than 90%. Adult emergences of the progeny of eggs stored at ultra-low temperatures allowed us to establish a laboratory colony.

Topics: Animals; Cryopreservation; Cryoprotective Agents; Dehydration; Female; Male; Moths; Ovum; Sodium Hypochlorite

The effects of temperature and salinity on the embryonation period and hatching success of eggs of Benedenia seriolae were investigated. Temperature strongly influenced embryonation period; eggs first hatched 5 days after laying at 28 degrees C and 16 days after laying at 14 degrees C. The relationship between temperature and embryonation period is described by quadratic regression equations for time to first and last hatching. Hatching success was >70% for B. seriolae eggs incubated at temperatures from 14 to 28 degrees C. However, no B. seriolae eggs embryonated and hatched at 30 degrees C and <2% of eggs hatched when incubated at 24 degrees C after transfer to 30 degrees C for 48 h. Embryonation period was similar for eggs incubated in sea water at 25, 30 and 35 per thousand salinity, but increased for eggs incubated at higher or lower salinities. When incubated at salinities ranging from 25 to 45 per thousand, more than 70% of B. seriolae eggs embryonated and hatched. Hatching success was lower at 20 and 50 per thousand salinity and few or no eggs hatched at 10 and 15 per thousand. Hatching of B. seriolae eggs can be prevented by desiccation for 3 min, by immersion in water at 50 degrees C for 30 s or by treatment with 25% ethanol for 3 min.

Topics: Analysis of Variance; Animals; Dehydration; Embryo, Nonmammalian; Ethanol; Hydrogen Peroxide; Perciformes; Platyhelminths; Seawater; Sodium Chloride; Sodium Hypochlorite; Temperature; Time Factors