Inactivation Of Bacillus Spores Research Articles

Comparison of Synergistic Effect of Nisin and Monolaurin on the Inactivation of Three Heat Resistant Spores Studied by Design of Experiments in Milk

Spore forming bacteria are special problems for the dairy industry. Heat treatments are insufficient to kill the spores. This is a continuously increasing problem for the industry, but we should be able to control it. In this context, we investigated the combined effect of nisin, monolaurin, and pH values on three heat resistant spores in UHT milk and distilled water and to select an optimal combination for the maximum spore inactivation. The inhibitory effect of nisin (between 50 and 200 IU/ml), monolaurin (ranging from 150 to 300 µg/ml), and pH (between 5 and 8) was investigated using a central composite plan. Results were analyzed using the response surface methodology (RSM). The obtained data showed that the inactivation of Bacillus spores by the combined effect of nisin-monolaurin varies with spore species, acidity, and nature of the medium in which the bacterial spores are suspended. In fact, Terribacillus aidingensis spores were more resistant, to this treatment, than Paenibacillus sp. and Bacillus sporothermodurans ones. The optimum process parameters for a maximum reduction of bacterial spores (∼3log) were obtained at a concentration of nisin >150 IU/ml and of monolaurin >200 µg/ml. The current study highlighted the presence of a synergistic effect between nisin and monolaurin against heat bacterial spores. So, such treatment could be applied by the dairy industry to decontaminate UHT milk and other dairy products from bacterial spores.

Biological Validation of a Chemical Effluent Decontamination System

Introduction: Failure of an existing effluent decontamination system (EDS) prompted the consideration of commercial off-the-shelf solutions for decontamination of containment laboratory waste. A bleach-based chemical EDS was purchased to serve as an interim solution. Methods: Studies were conducted in the laboratory to validate inactivation of Bacillus spores with bleach in complex matrices containing organic simulants including fetal bovine serum, humic acid, and animal room sanitation effluent. Results: These studies demonstrated effective decontamination of >106 spores at a free chlorine concentration of ≥5700 parts per million with a 2-hour contact time. Translation of these results to biological validation of the bleach-based chemical EDS required some modifications to the system and its operation. Discussion: The chemical EDS was validated for the treatment of biosafety levels 3 and 4 waste effluent using laboratory-prepared spore packets along with commercial biological indicators; however, several issues and lessons learned identified during the process of onboarding are also discussed, including bleach product source, method of validation, dechlorination, and treated waste disposal.

Inactivation of Bacillus Spores in Wash Waters Using Dilute Chlorine Bleach Solutions at Different Temperatures and pH Levels.

Inactivation of Bacillus globigii spores in wash water was studied to simulate chlorine inactivation of Bacillus anthracis spores in water generated during biological cleanups. Eight waters were studied, with six containing detergent. Chlorine levels were approximately 3000 mg/L. Results across different waters showed decreasing inactivation with increasing pH. Inactivation did not appear to be influenced by chemical oxygen demand, suspended solids, turbidity, or dissolved solids. Inactivation efficacy was expressed as the time calculated to yield 6 log10 inactivation at 3000 mg NaOCl/L. This time ranged from 5 to 51 minutes at ~21 °C and from 11 to 209 minutes at ~5 °C. For one wash water, inactivation was conducted when there was no pH adjustment, and when the pH was buffered at 7 and 8. Inactivation in these buffered waters was rapid, but inactivation decreased sharply at a pH above ~9.3.

Inactivation ofBacillusspores in decontamination wash down wastewater using chlorine bleach solution

Bacillus anthracis spores are highly resistant to disinfection. The effectiveness of sodium hypochlorite (bleach) solution for the inactivation of Bacillus atrophaeus subspecies globigii spores (a surrogate for Bacillus anthracis) in wash down wastewater was evaluated by a series of bench-scale and pilot-scale experiments. The wastewater was generated from a simulated decontamination of interior laboratory surfaces and personal protective equipment. A concentration of approximately 5% (v/v) bleach solution was adequate for a ≥7 log10inactivation of Bacillus spores spiked in wastewater after 10 min of contact time at room temperature. This sodium hypochlorite dose is less than the U.S. National Response Team-recommended formulation of 10% (v/v) bleach solution with 10% (v/v) vinegar. For the wash down waters evaluated, the addition of vinegar was not required. Inactivation of Bacillus spores was faster when a detergent was added to the wash down waters tested.

Influence of the surface temperature of packaging specimens on the inactivation of Bacillus spores by means of gaseous H2O2

To determine the influence of condensation as a function of the surface temperature of aseptic packaging, on the inactivation of Bacillus spores [Bacillus subtilis (DSM 347), B. subtilis SA22, Bacillus atrophaeus] having different surface properties by means of vaporized H(2) O(2). The packaging specimens inoculated with Bacillus spores were tempered and subsequently exposed to H(2) O(2) -vapour. During the exposure, surface temperature curves were measured and the spore survival was determined. Results showed that decreasing the initial surface temperature of the packaging specimens had a positive effect on the sporicidal activity of H(2) O(2) -vapour, where the effect was less pronounced for less hydrophilic spores. The surfaces of spores were characterized by means of the water contact angle. For starting surface temperatures below the dew point temperature of the sterilant gas, the condensation of highly concentrated liquid H(2) O(2) on the packaging surface accelerates the killing of the spores, while the inferior wettability of more hydrophobic spores compared to more hydrophilic ones diminishes the effect. Regarding industrial packaging sterilization, a mixed microflora has to be inactivated. Promoting the condensation of H(2) O(2) improves in general the killing of different species of spores, however, at various degrees depending on the wettability of spores.

Hydrostatic Pressure-Induced Germination and Inactivation of Bacillus Spores in the Presence or Absence of Nutrients

The influence of pressure and temperature, in the presence (in glucose broth, GB) or absence (in phosphate buffer, PB) of nutrients, on hydrostatic pressure (HP) induced-germination and inactivation of Bacillus subtilis, B. cereus, and B. polymyxa spores was investigated for a relatively low range of 20-100MPa (low HP). Low HP-induced germination was more potent in GB than in PB at 20-60MPa, and the difference in germination ratio between the growth media (GB and PB) tended to decrease with increasing pressure. For B. subtilis and B. cereus, the optimal temperature for the low HP-induced germination in PB tended to be higher than in GB, whereas temperature dependence on low HP-induced germination of B. polymyxa was different considerably from those of B. subtilis and B. cereus. After 30-60min incubation at 40°C and 60MPa, the three types of bacterial spores germinated 5 log-cycles in the presence of nutrients and 2-3 log-cycles in the absence of nutrients. These results indicate that the low HP-induced germination of Bacillus spores has potential as a method of reducing bacterial spores in foods and foodstuffs, especially those containing large amounts of nutrients.

Inactivation of Bacillus spores in reconstituted skim milk by combined high pressure and heat treatment

To determine the resistance of a variety of Bacillus species spores to a combined high pressure and heat treatment; and to determine the affect of varying sporulation and treatment conditions on the level of inactivation achieved. Spores from eight Bacillus species (40 isolates) were high pressure-heat treated at 600 MPa, 1 min, initial temperature 72 degrees C. The level of inactivation was broad (no inactivation to 6 log10 spores ml(-1) reduction) and it varied within species. Different sporulation agar, high pressure equipment and pressure-transmitting fluid significantly affected the response of some isolates. Varying the initial treatment temperature (75, 85 or 95 degrees C) shifted the relative order of isolate high pressure-heat resistance. The response of Bacillus spores to combined high pressure-heat treatment is variable and can be attributed to both intrinsic and extrinsic factors. The combined process resulted in a high level of spore inactivation for several Bacillus species and is a potential alternative treatment to traditional heat-only processes. Sporulation conditions, processing conditions and treatment temperature all affect the response of Bacillus spores to the combined treatment of high pressure and heat. High levels of spore inactivation can be achieved but the response is variable both within and between species.

Inactivation of Bacillus spores by the combination of moderate heat and low hydrostatic pressure in ketchup and potage

The combination effect of moderate heat and low hydrostatic pressure (MHP) on the reduction of Bacillus subtilis, Bacillus coagulans and Geobacillus stearothermophilus spores in food materials (potage and ketchup) was investigated. These bacterial spores were suspended in potage (pH 7), acidified potage (pH 4), neutralized ketchup (pH 7) and ketchup (pH 4). The suspensions were treated with and without pressure (100 MPa) and temperatures of 65–85 °C for 3 to 12 h. The bacterial spores were inactivated by 4–8 log cycles during MHP treatment in potage, acidified potage and ketchup, whereas the spores were highly resistant to long time heat treatment in potage and neutralized ketchup. The degrees of spore destruction were mostly dependent on pH and medium composition during MHP treatment. The inactivation effect in MHP treatment was higher at the pH 7 than at pH 4 both in ketchup and potage. The bacterial spores showed higher inactivation in potage than ketchup during MHP treatment.

Inactivation of Bacillus spores by gaseous ozone.

The sporicidal activity of ozone in the gas phase was investigated. Spores of six strains of Bacillus species deposited on filter paper or glass fibre filter were conditioned at different relative humidities (r.h.), and then exposed to ozone ranging in concentration from 0.5 to 3.0 mg/l at different r.h. There was a lag phase in the initial stage of exposure followed by an exponential decrease in the number of survivors with time, although no lag phase was observed with one strain. Inactivation rates increased with increasing exposure r.h. while no significant inactivation was attained at a r.h. of 50% or below. The conditioning r.h. influenced the duration of the lag phase. The D-values (decimal reduction time) in the logarithmic phase varied roughly in inverse proportion to the ozone concentration.

Inactivation of Bacillus Spores in Dry Systems at Low and High Temperatures

A plot of the thermal resistance of Bacillus subtilis var. niger spores (log D value) against temperature was linear between 37 and 190 degrees C (z = 23 degrees C), provided that the relative humidity of the spore environment was kept below a certain critical level. The corresponding plot for Bacillus stearothermophilus spores was linear in the range 150 to 180 degrees C (z = 29 degrees C) but departed from linearity at lower temperatures (decreasing z value). However, the z value of 29 degrees C was decreased to 23 degrees C if spores were dried before heat treatment. The straight line corresponding to this new z value was consistent with the inactivation rate at a lower temperature (60 degrees C). The data indicate that bacterial spores which are treated in dry heat at an environmental relative humidity near zero are inactivated mainly by a drying process. By extrapolation of the thermal resistance plot obtained under these conditions for B. subtilis var. niger spores, the D value at 0 degrees C would be about 4 years.