An impedimetric method for rapid screening of cosmetic preservatives

An efficient impedance method was developed for rapid evaluation of cosmetic preservatives. The method used decimal reduction time or D-value to assess preservative efficacies. The D-value, which was calculated from the plot of Log CFU ml^–1 versus time by linear regression analysis, could be obtained within 48 h. Thus, the time required for the challenge test was reduced from 4–8 weeks with the standard procedures (eg US Pharmacopeia), to 2 days with the current method. A calibration curve (r=-0.95) was established by plotting the Log CFU ml^–1 versus capacitance detection time (DT) of 108 samples. With the calibration, CFU can be estimated directly from the impedance test without plating. Two commercial biocides and several other chemicals were evaluated in a shampoo by the impedance procedure againstPseudomonas aeruginosa. The D-values obtained from the impedance test were not significantly different from those produced by the conventional plate count method. The technique was found to be particularly useful when screening a large number of compounds to find novel preservatives and synergistic preservative combinations.     

Effect of environmental stresses on the sensitivity of Enterobacter sakazakii in powdered infant milk formula to gamma radiation

Aim:  To evaluate the effect of starvation, heat, cold, acid, alkaline, chlorine and ethanol stresses on the resistance of Enterobacter sakazakii in powdered infant milk formula (PIMF) towards gamma radiation.
Methods and Results:  Stressed cells of E. sakazakii ATCC 51329 and four other food isolate strains were mixed individually with PIMF, kept overnight at room temperature, and then exposed to gamma radiation up to 7·5 kGy. The D ₁₀-values were determined using linear regression and for the stressed E. sakazakii strains these values ranged from 0·82 to 1·95 kGy.
Conclusions:  Environmental stresses did not significantly change the sensitivity of most E. sakazakii strains to ionizing radiation.
Significance and Impact of the Study:  Data obtained established that most forms of environmental stress are unlikely to significantly enhance the resistance of E. sakazakii strains to lethal, low dose irradiation treatment.     

Interaction of pressure, time and temperature of pressurization on viability loss of Listeria innocua

In this study, the combined effectiveness of pressures of 137.9–344.7MPa, temperatures of 25–50°C and exposure times of 5–15min on Listeria innocua viability in peptone solution is examined. The results showed that under the study conditions only the combination of 344.7MPa, 50°C and 9.1min can reduce the viability of this species by 7 logs, with a z value of 173.1MPa.     

The growth profile, thermotolerance and biofilm formation of Enterobacter sakazakii grown in infant formula milk

AIMS: To study the growth, thermotolerance and biofilm formation of the emergent pathogen Enterobacter sakazakii in infant formula milk (IFM). METHODS AND RESULTS: The temperature range, death kinetics and biofilm formation of E. sakazakii were determined using impedance microbiology and conventional methods. In IFM the organism grew as low as 6 degrees C and optimally at 37-43 degrees C. In faecal coliform tests, 23% of strains (n = 70) produced gas from lauryl sulphate broth (LSB) at 44 degrees C after 48 h incubation. Three strains failed to grow in LSB at any of the temperatures. The D-value of cells suspended in IFM was determined between 54 and 62 degrees C. The resultant z-value was 5.7 degrees C. The organism was able to adhere and grow on latex, polycarbonate, silicon and to a lesser extent stainless steel. CONCLUSIONS: Enterobacter sakazakii was able to grow at refrigeration temperatures and on infant-feeding equipment. The thermotolerance of the organism was similar to other Enterobacteriaceae and should be killed during standard pasteurization treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: Enterobacter sakazakii has been associated with infant meningitis through consumption of contaminated IFM. Enterobacter sakazakii is able to grow in IFM during storage at refrigeration temperatures and attach to infant-feeding equipment, which may become reservoirs of infection.     

Using Thermal Inactivation Kinetics to Calculate the Probability of Extreme Spore Longevity: Implications for Paleomicrobiology and Lithopanspermia

Thermal inactivation kinetics with extrapolation were used to model the survival probabilities of spores of various Bacillus species over time periods of millions of years at the historical ambient temperatures (25–40 °) encountered within the 250 million-year-old Salado formation, from which the putative ancient spore-forming bacteriumSalibacillus marismortui strain 2-9-3 was recovered. The model indicated extremely low-to-moderate survivalprobabilities for spores of mesophiles, but surprisingly high survival probabilities for thermophilic spores. The significance of the results are discussed in terms of the survival probabilities of (i) terrestrial spores in ancient geologic samples and (ii) spores transported betweenplanets within impact ejecta.     

Effects of superheated steam on Geobacillus stearothermophilus spore viability

Aims: To examine the effect of processing with superheated steam (SS) on Geobacillus stearothermophilus ATCC 10149 spores. Methods and Results: Two inoculum levels of spores of G. stearothermophilus were mixed with sterile sand and exposed to SS at 105–175°C. The decimal reduction time (D‐value) and the thermal resistance constant (z‐value) were calculated. The effect of cooling of spores between periods of exposure to SS was also examined. A mean z‐value of 25·4°C was calculated for both inoculum levels for SS processing temperatures between 130°C and 175°C. Conclusions: Spore response to SS treatment depends on inoculum size. SS treatment may be effective for reduction in viability of thermally resistant bacterial spores provided treatments are separated by intermittent cooling periods. Significance and Impact of the Study: There is a need for technologies that require short thermal processing times to eliminate bacterial spores in foods. The SS processing technique has the potential to reduce microbial load and to modify food texture with less energy in comparison to commonly used hot air treatment. This work provides information on the effect of SS processing parameters on the viability of G. stearothermophilus spores.     

Destruction and injury of Escherichia coli during microwave heating under vacuum

AIMS: To study the effect of 2450 MHz microwave radiation under vacuum (vacuum microwave or VM) on survival and injury of Escherichia coli and to search for possible nonthermal effects associated with VM. METHODS AND RESULTS: Destruction kinetics of E. coli in peptone water were determined in a continuous-flow vacuum system, heated by convection heating in a water bath or with microwaves (VMs). Vacuum was used to control the boiling point of water and to maintain temperature in the bacterial suspensions at specified levels (49-64 degrees C). CONCLUSIONS: z-Value in the water bath treatment was 9.1 degrees C while for VM at 510 and 711 W it was 6.2 and 5.9 degrees C, suggesting that E. coli is more sensitive to temperature changes under microwave heating. Arrhenius calculations of the activation energies of the destruction reactions suggest that the mechanism of destruction in VM may be different from that of conventional heat. The number of injured micro-organisms showed no significant differences among treatments. SIGNIFICANCE AND IMPACT OF THE STUDY: The impact of temperature on E. coli destruction was different when microwaves were the medium of heat transfer, suggesting the existence of factors other than heat contributing to the lethal effect of VM.     

Validation of a polynomial regression model: the thermal inactivation of Bacillus subtilis spores in milk

AIMS: The predicted survival of Bacillus subtilis 168 spores from a polynomial regression equation was validated in milk. METHODS AND RESULTS: Bias factor suggested as an index of model performance was used to validate the polynomial model predictions in ultrahigh temperature (UHT) treated and sterilized whole and skim milk. Model predictions were fail safe, predicting higher D-values (decimal reduction times) in buffer than actually noted in milk. CONCLUSIONS: The D-values for spores were lower in milk as compared with those predicted in potassium phosphate buffer contrary to the popular expectation of better spore survival in complex food systems. The Bias factor, a quantitative measure of the model performance, indicated that on average the model predictions exceed the observations by 40% in the case of whole milk and by 60% in the case of skim milk. SIGNIFICANCE AND IMPACT OF THE STUDY: The present work is an attempt to ascertain the extent of reliability that one can safely place in polynomial model predictions, without compromising on the safety or palatability of foods where it is eventually intended to be applied. The work has also highlighted the differences in the thermal inactivation pattern of spores in buffer and in milk with a possible influence of the various constituents of milk. The work will assist the dairy industry to better design thermal processes to ensure longer shelf life of dairy foods.     

两种天然植物提取物的抑菌效果的快速评价与比较

本文研究了绿茶和桑叶提取物在不同浓度时对细菌的抑制作用,采用D值计算法快速评价了它们的抑菌效果。结果表明,绿茶提取物对金黄色葡萄球菌和大肠杆菌的最低抑制浓度分别为0.034%,0.42%;而桑叶提取物分别为0.86%,1.54%。绿茶的抑菌效果更好。     

Thermal characteristics of recombinant green fluorescent protein (GFPuv) extracted from Escherichia coli

AIMS: The thermal stability of isolated and extracted recombinant green fluorescent protein (GFPuv) was evaluated by analysing the loss of fluorescence intensity. METHODS AND RESULTS: GFPuv was expressed by Escherichia coli, extracted by the three-phase partitioning method and purified by elution through an hydrophobic interaction column. The collected fractions were further diluted in Tris-HCl-EDTA (pH 8.0) and subjected to continuous heating at set temperatures (45-95 degrees C). From a standard curve relating fluorescence intensity to GFPuv concentration, the loss of fluorescence intensity was converted to denatured GFPuv concentration (microg ml-1). To determine the extent of the thermal stability of GFPuv, decimal reduction times (D-values), z-value and energy of activation (Ea) were calculated. CONCLUSIONS: For temperatures between 45 and 70 degrees C, extracted native GFPuv activity decreased from 11 to 75% relative to initial native protein concentration above 70 degrees C, the average decrease in GFPuv fluorescence was between 72 to 83%. SIGNIFICANCE AND IMPACT OF THE STUDY: The thermal stability of GFPuv provides the basis for its potential utility as a fluorescent biological indicator to assess the efficacy of the treatment of liquids and materials exposed to steam.