(-)-Tetrahydroberberrubine∙acetate accelerates antioxidant potential and inhibits food associated Bacillus cereus in rice.

A protoberberine alkaloid, (-)-tetrahydroberberrubine∙acetate (THBA) was assessed for its antioxidant potential and ability to inhibit the growth of a food hazard bacterium Bacillus cereus in vitro and in situ. THBA displayed significant and dose-dependent cellular antioxidant potential against hydrogen peroxide-induced oxidative stress in NIH 3T3 fibroblast cells and decreased the ROS levels as well as increased the expression levels of SOD1 and SOD2 enzymes. The inhibitory spectrum of THBA confirmed its mechanistic role in the disruption of the membrane integrity of B. cereus as evidenced by the results of time-inactivation, cell membrane integrity, NPN membrane uptake, membrane potential, and electron microscopy analyses. Moreover, THBA inhibited biofilm formation by B. cereus and disrupted pre-established biofilms on a glass surface. Furthermore, THBA was also able to inhibit B. cereus in raw rice with a significant amount of reduction in CFU counts, suggesting its potential role as a natural antioxidant and antimicrobial agent.

Flow path system of ultraviolet C irradiation from xenon flash to reduce bacteria survival in platelet products containing a platelet additive solution.

BACKGROUND: Our previous study showed that ultraviolet C (UVC) from xenon (Xe) flash without any photoreactive compounds inactivated bacteria in platelet concentrates (PCs) with less damage to platelets (PLTs) as compared with Xe flash containing ultraviolet A, ultraviolet B, and visible light. Here, we report a UVC irradiation system for PCs under flow conditions consisting of a flow path-irradiation sheet, a peristaltic pump, and a collection bag. STUDY DESIGN AND METHODS: Platelet concentrates containing Ringer's solution (R-PCs) inoculated with bacteria were injected into a flow path sheet using a peristaltic pump, being irradiated with UVC from Xe flash. The quality of the irradiated PCs containing platelet additive solution (PAS-PCs) was assessed based on PC variables, PLT surface markers, and aggregation ability. RESULTS: Streptococcus dysgalactiae (12 tests) and Escherichia coli (11) were all negative on bacterial culture, while Staphylococcus aureus (12) and Klebsiella pneumoniae (14) grew in one and two R-PCs, respectively. Bacillus cereus spores were inactivated in 7 of 12 R-PCs. PC variables became significantly different between irradiated and nonirradiated PAS-PCs. P-selectin, first procaspase-activating compound (PAC-1) binding, and phosphatidylserine increased by irradiation. Aggregability stimulated by adenosine diphosphate, collagen, or thromboxane A2 increased in the irradiated PAS-PCs, while that by thrombin became smaller compared with nonirradiated controls. CONCLUSION: This newly developed system inactivated bacteria including spores in R-PCs. PAS-PCs irradiated by this system retained acceptable in vitro quality and aggregability. Usage of a peristaltic pump instead of agitator during irradiation may enable this system to be directly combined with an apheresis blood cell separator.

Inactivation of Bacillus and Clostridium Spores in Coconut Water by Ultraviolet Light.

Bacterial spores are generally more resistant than vegetative bacteria to ultraviolet (UV) inactivation. The UV sensitivity of these spores must be known for implementing UV disinfection of low acid liquid foods. UV inactivation kinetics of bacterial spores in coconut water (CW) and distilled sterile water was studied. Populations of Bacillus cereus and Clostridium sporogenes dormant spores were reduced by more than 5.5 log10 at the UV-C photon fluence of 1142 µE·m-2 and 1919 µE·m-2 respectively. C. sporogenes spores showed higher UV-C resistance than B. cereus, with the photon fluence 300 µE·m-2 required for one log inactivation (D10) and 194 µE·m-2, respectively. No significant difference was observed in D10 values of spores suspended in the two fluid types (p > 0.05). The inactivation kinetics of microorganisms were described by log linear models with low root mean square error and high coefficient of determination (R2 > 0.98). This study clearly demonstrated that high levels of inactivation of bacterial spores can be achieved in CW. The baseline data generated from this study will be used to conduct spore inactivation studies in continuous flow UV systems. Further proliferation of the technology will include conducting extensive pilot studies.

Decontamination of Mesquite Pod Flour Naturally Contaminated with Bacillus cereus and Formation of Furan by Ionizing Irradiation.

Mesquite pod flour produced from nitrogen-fixing trees of the Prosopis species has a unique aroma and flavor that is preferred by some consumers. Due to the presence of wildlife, grazing domestic animals, and insects, the pods have a high potential of being contaminated with human pathogenic bacteria, such as Bacillus cereus. Nonthermal processing technologies are helpful to reduce the population of microorganisms in the flour because heating deteriorates the characteristic flavor. A study was conducted to investigate the efficacy of ionizing radiation in decontaminating two types of mesquite pod flours (Prosopis alba and Prosopis pallida) naturally contaminated with B. cereus and the effects of irradiation on the formation of furan, a possible human carcinogen. Results showed that the populations of B. cereus were 3.8 and 5.4 log CFU/g in nonirradiated P. alba and P. pallida flours, respectively, and populations of microflora, mesophilic spores, B. cereus, and B. cereus spores decreased with increasing radiation doses. At 6 kGy, the populations fell below 1 log CFU/g. Irradiation at 6 kGy had no significant effect on the fructose, glucose, or sucrose content of the flour. Nonirradiated P. alba and P. pallida flours contained 13.0 and 3.1 ng/g of furan, respectively. Furan levels increased with irradiation doses at rates of 2.3 and 2.4 ng/g/kGy in the two flours. The level of 3-methylbutanal was reduced or not affected by irradiation, while the hexanal level was increased. Our results suggested that irradiation was effective in decontaminating contaminated mesquite flour. The significance of furan formation and possible changes in flavor due to irradiation may need to be further examined.

Combined effect of cadmium, lead, and UV rays on Bacillus cereus using comet assay and oxidative stress parameters.

Exposure to environmental chemicals and oxidative stress particularly at low dose levels may produce additive or synergistic interactions not seen in single component exposure. Exposure to cadmium, lead, and ultraviolet rays occurs in many occupational settings, such as pigment and battery production, galvanization, and recycling of electric tools. However, little is known about interactions between heavy metals and ultraviolet rays. This study aimed to evaluate the interactions of ultraviolet rays of 254 nm (UV-B) with cadmium or lead on Bacillus cereus. B. cereus was treated with different concentrations of cadmium or lead followed by exposure to UV-B radiation as combined effect. Photoirradiation of B. cereus with UV-B with exposure to cadmium or lead results in DNA damage, cytotoxicity, depletion of glutathione, and formation of lipid peroxidation. UV-B rays alone enhanced glutathione production which was depleted with lead and high doses of cadmium. Lead alone does not increase DNA breaking. The mechanism behind these interactions might be repair inhibition of oxidative DNA damage, since a decrease in repair capacity will increase susceptibility to reactive oxygen species generated by cadmium or lead. Lipid peroxidation was increased with exposure to UV-B and cadmium or lead. DNA, glutathione, and lipid peroxidation can be used as biomarkers to identify possible environmental contamination in bacteria. One conclusion from this model is the existence of more than multiplicative effects for co-exposures of cadmium or lead and UV rays.

Synergistic effects of NaOCl and ultrasound combination on the reduction of Escherichia coli and Bacillus cereus in raw laver.

The current study investigated the synergistic effects of NaOCl (50-200 ppm)/ultrasound (37 kHz, 380 W for 5-100 min) combination on the reduction of Escherichia coli and Bacillus cereus in raw laver. The synergistic reductions were not dependent on concentrations of NaOCl and times of ultrasound. Synergistic reduction ranged from 0.1 to 0.5 log10 colony-forming units (CFU)/g and 0.1-1.1 log10 CFU/g, respectively, for E. coli and B. cereus, with the largest synergistic reduction in the combination of 200 ppm NaOCl and 60-min ultrasound. Moreover, significant differences of "L" (lightness), "a" (redness), and "b" (yellowness) were not observed in combined with 50-200 ppm NaOCl and 100-min ultrasound compared to those in raw laver treated by only 100-min ultrasound. The results in the current study indicate that the combined treatment of 200 ppm NaOCl and 60-min ultrasound could be regarded a potential optimum hurdle approach in the seaweed production, processing, and distribution process to enhance seaweed safety.

Response of Bacillus cereus vegetative cells after exposure to repetitive sublethal radiation processing in combination with nisin.

The present study evaluated the response of Bacillus cereus LSPQ 2872 vegetative cells, following exposure to single and repetitive sublethal γ-radiation treatment at 1 kGy alone or in combination with nisin at its maximum tolerated concentration, in BHI broth supplemented with 0.5% glucose. Results showed that B. cereus has the capability to develop increased resistance to subsequent cycles of gamma irradiation and a significant increase (p ≤ 0.05) of the relative radiation resistance (D(10)) value was noticed after the fourth irradiation at 1 kGy. The sequence and the repetitive sublethal treatment of γ-radiation with nisin affected significantly (p ≤ 0.05) B. cereus radio-tolerance since lower D(10) values were recorded. Our results showed that the response of B. cereus to repetitive mild bactericidal treatment was accompanied by relevant modifications in the cell properties leading to the increased resistance. Transmission electron microscopy revealed that resistance might be related to changes in the cell wall. Multiparameter flow cytometry revealed effectively diverse physiological changes that B. cereus underwent during development of radio-resistance. The development of increased resistance to gamma irradiation was accompanied by an increase of the percentage of injured and viable cells at the expense of dead cells. Nisin treated cells developed also increased radio-resistance if repetitively processed with γ-radiation at sublethal doses which was not demonstrated by plate counting.

Synergistic effects of sodium hypochlorite and ultraviolet radiation in reducing the levels of selected foodborne pathogenic bacteria.

The purpose of this study was to determine whether combined treatment would produce synergistic effects to facilitate the sterilization of food products during production relative to single treatment. To assess this hypothesis, we investigated the bactericidal effects of ultraviolet (UV) irradiation and a commercial chemical disinfectant, sodium hypochlorite (NaClO), on Bacillus cereus F4810/72, Cronobacter sakazakii KCTC 2949, Staphylococcus aureus ATCC 35556, Escherichia coli ATCC 10536, and Salmonella Typhimurium novobiocin/nalidixic acid in vitro. Various concentrations of NaClO (20, 60, 100, and 200 ppm NaClO) were tested along with exposure to UV radiation at various doses (6, 96, 216, 360, and 504 mW s/cm(2)). The combined NaClO/UV treatments resulted in greater reductions in bacterial counts than either treatment alone. The synergy values against B. cereus, C. sakazakii, S. aureus, Salmonella Typhimurium, and E. coli were 0.25-1.17, 0.33-1.97, 0.42-1.72, 0.02-1.44, and 0.01-0.85 log(10) CFU/mL, respectively. The results of this study suggest that a significant synergistic benefit results from combined NaClO/UV processing against food-borne pathogenic bacteria in vitro.

Response of Bacillus cereus to gamma-irradiation in combination with carvacrol or mild heat treatment.

Carvacrol and mild heat treatment were tested for their efficiency to increase the radiosensitivity of Bacillus cereus in broth. The bacterium was treated with gamma-irradiation alone or in combination with carvacrol at its minimal inhibitory concentration or mild heat treatment for 10 min at 45 degrees C. The effects of this combination of treatments were studied on various parameters: the bacterial viability, the modifications of the cell morphology with scanning electron microscopy (SEM), the cellular fatty acids composition of the membrane quantified by gas chromatography, the intracellular and extracellular adenosine 5'-triphosphate (ATP) concentrations, and the DNA degradation. Combined treatments resulted in additive or synergistic effects as compared to gamma-irradiation alone. A significant modification (P < or = 0.05) of the fatty acid composition and unsaturation ratios was observed. Pretreatment with mild heat or carvacrol before irradiation disturbed the membrane integrity of B. cereus and induced a significant decrease (P < or = 0.05) of the intracellular ATP concentration. SEM observations revealed that the cell membrane was more severely affected with combined treatment than irradiation alone. The electrophoresis analysis showed that DNA degradation by combined treatments was greater than the gamma-irradiation alone.

Broadband 10-300 GHz stimulus-response sensing for chemical and biological entities.

By illuminating the sample with a broadband 10-300 GHz stimulus and coherently detecting the response, we obtain reflection and transmission spectra of common powdered substances, and compare them as a starting point for distinguishing concealed threats in envelopes and on personnel. Because these samples are irregular and their dielectric properties cannot be modulated, however, the spectral information we obtain is largely qualitative. To show how to gain quantitative information on biological species at micro- and millimetre-wave frequencies, we introduce thermal modulation of a globular protein in solution, and show that changes in single-wavelength microwave reflections coincide with accepted visible absorption spectra, pointing the way towards gaining quantitative chemical and biological spectra from broadband terahertz systems.

Radiation sensitivity of Bacillus cereus with and without a crystalline surface protein layer.

The radiation sensitivity of four strains of Bacillus cereus was investigated with attention to bacterial surface structure. All four strains were sensitive to radiation with gamma rays (D(10)=0.4 kGy). No crystalline surface protein layer could be detected on the cell surface. When cultured on solid media, an S-layer covered the cells of the two strains, and they were 2.6 times as resistant to radiation as the two reference strains without an S-layer. In SDS-PAGE, a major 97-kDa band from the resistant strains from plate cultures was replaced by a ca. 85-kDa protein band in samples from broth cultures. Electron microscopy, SDS-PAGE, Western blot and fluorescent antibody staining indicated that the higher resistance to radiation of the clinical strains from plate cultures was associated with the presence of the S-layer on the cell surface.

Radiation inactivation of some food-borne pathogens in fish as influenced by fat levels.

The influence of low (0.39-1.1%), medium (4.25%) and high (7.1-32.5%) fat levels in fish on radiation inactivation of four food-borne pathogens was investigated. Cells of Listeria monocytogenes 036, Yersinia enterocolitica F5692, Bacillus cereus and Salmonella typhimurium at logarithmic phase were inoculated in 10% fish homogenates and subjected to gamma irradiation at ice temperature (0-1 degree C) with doses ranging from 0.05 to 0.8 kGy. The radiation survival curves of L. monocytogenes and B. cereus were characterized by shoulders, while a tailing effect was depicted by cells of Y. enterocolitica and B. cereus. The D10 values in kGy calculated on the exponential part of the curve ranged from 0.2 to 0.3, 0.15 to 0.25, 0.1 to 0.15 and 0.09 to 0.1 for L. monocytogenes 036, B. cereus, Salm. typhimurium and Y. enterocolitica F5692, respectively. This order (D10) of radiation resistance of each organism was not affected by the fat content of the fish. Inoculated pack studies carried out separately with each pathogen in fatty (Indian sardine, 7.1%) and lean (Golden anchovy, 0.39%) fish showed no difference in their survival after exposure to 1 kGy and 3 kGy doses, which corroborated the above observation. The practical significance of these results in the application of the technology is discussed.

Efecto de las microondas sobre la sobrevivencia de bacterias esporuladas inoculadas en carne molida / Microwave effect on the survival of sporulated bacteria inoculated in minced meat

Debido a la creciente tendencia de cocinar y recalentar en horno de microondas platillos a base de carne y a la posibilidad de que estos transmitan enfermedades de origen bacteriano, se evaluó la capacidad de sobrevivencia de esporas de bacterias inoculadas en carne molida y tratadas en horno de microonda. Las tortas de carne fueron inoculadas con un número conocido de esporas de Bacillus cereus o Clostridium perfringens y posteriormente cocinadas en horno de microondas Amana de 2450 Hz. Se utilizó la técnica descrita por Vanderzant y Splittstoesser para determinar la tase de sobrevivencia y la actividad de la enxima fosfatasa ácida como parámetro de conocimiento. Las esporas de B. cereus mostraron una tendencia a disminuir en número, conforme aumenta el tiempo de exposición a la radiación, pero si llegar a desaparecer totalmente. Las esporas de C. perfringens disminuyen en número al aumentar el tiempo de exposición, pero presentan un segundo aumento asociado a la germinación de esporas sobrevivientes.

[A comparative study of ethylene oxide and ionizing radiation for sterilizing bone grafts].

To find a good way for sterilization and disinfection of bone grafts, we compared the sterilization capacity of gaseous ethylene oxide (EO) and cobalt-60 gamma radiation. The bone chips were contaminated with 10(7) bacteria per milliliter of Staphylococcus aureus ATCC 25923, Bacillus subtilis globigii 8017 and Bacillus cereus 4001, then sterilized with various doses of gaseous EO or cobalt- 60 gamma radiation. The sterilization effect of EO was more stronger and faster than that of 60Co gamma radiation. The application of moderate doses of EO for sterilizing particulate bone grafts was recommended.

Effect of low-dose irradiation on growth of and toxin production by Staphylococcus aureus and Bacillus cereus in roast beef and gravy.

The effect of irradiation (2 kGy) on growth of and toxin production by Staphylococcus aureus and Bacillus cereus in roast beef and gravy during storage at abuse temperatures (15 and 22 degrees C) was assessed by inoculation studies. Irradiation resulted in a 3-4 log10 reduction in numbers of both pathogens. Whenever B. cereus and S. aureus numbers reached 10(6) and 10(7) cfu/g, respectively, during storage their toxins were detectable. As the time taken to attain these levels was longer in irradiated than in unirradiated samples, toxin production by both pathogens was delayed by irradiation. When samples initially containing low levels (10(2)/g) of S. aureus were irradiated no toxin was produced during subsequent storage at 15 or 22 degrees C. Diarrhoeal toxin produced by B. cereus was detected after 2 days at 22 degrees C, but not at 15 degrees C, in samples containing 10(2) cells/g prior to irradiation. When higher numbers (10(6)/g) of either pathogen were present prior to irradiation, toxins were produced by both pathogens at 22 degrees C, but not at 15 degrees C. Microbial competition had an effect on the growth of B. cereus and S. aureus after irradiation when a low initial inoculum was applied. However, when a higher inoculum was used the pathogens outnumbered their competitors and competition effects were less important. It was concluded that low-dose irradiation would improve the microbiological safety of roast beef and gravy.

Bactericidal action of carbon dioxide laser radiation in experimental dental root canals.

The ability of a carbon dioxide laser to sterilize the root canal of human teeth has been investigated. Three oral bacteria, Streptococcus sanguis, Streptococcus mutans, and Actinomyces viscosus, and three other bacteria, Bacillus cereus, Staphylococcus aureus, and Pseudomonas aeruginosa were used as experimental organisms. Exposure of cells on glass slides to laser radiation showed there was little difference in the exposure required to kill these six organisms. Complete recovery of bacteria from the root canal was initially a problem and was only achieved when bacterial manipulations and removal were carried out in rapid succession, within 5 min of inoculation. However, the geometry of the instrumented canal and the laser alignment were major factors in achieving consistent cell death of oral bacteria in the root canals. Using sets of 10 teeth, four repeated exposures of 10 W for 1 s was found to sterilize 4 or more of the teeth.

[Resistance of resting cyst-like forms of Bacillus cereus to exposure to high temperature, ultraviolet rays and low-molecular alcohols]. / Ustoichivost' pokoiashchikhsia tsistopodobnykh form Bacillus cereus k vozdeistviiu vysokoi temperatury, ul'trafioletovykh luchei i nizkomolekuliarnykh spirtov.

The dormancy cyst-like forms of Bacillus cereus obtained under the action of an autoregulatory factor were shown to be more resistant to the action of such external factors of the medium as high temperatures, ultraviolet irradiation and low molecular weight alcohols comparing to the vegetative cells. Incubation at 80 degrees C for 15 min killed 98,5% vegetative cells and 86,5% of the dormancy refractile forms. U. V.--radiation treatment resulted in a loss of viability in 97% of the vegetative cells and in 75% of the refractile forms. Differences were also found in the resistance of the cyst-like forms and the vegetative cells to the action of ethanol, methanol and butanol. The resistance of the cyst-like forms of B. cereus to the action of the above damaging factors is consistent, in general, with the resistance of cysts, methylotrophic bacterial spores and other dormancy prokaryotic cells differing in the mode of their formation from endospores.

Bacterial food-poisoning versus coliforms-faecal streptococci and gamma radiation survival/dose.

All the species tested were sensitive to gamma radiation treatment, even at a dose as low as 0.001 Mrad. The spore-former Bacillus cereus (PCJ 213) proved to be more resistant than the rest of the tested species, while the Gram-negative organisms were more susceptible to irradiation treatment. The tolerance among those organisms is depicted by the descending order: Proteus vulgaris, irregular VI, Escherichia coli I, E. coli III, and Salmonella typhi-murium. Yet all the tested faecal streptococci species, with the exception of Streptococcus bovis, may be regarded as holding an intermediate position between the more resistant B. cereus (PCJ 213) and the more sensitive Gram-negative organisms.