Antimicrobial activity of Enterococcus faecium L50, a strain producing enterocins L50 (L50A and L50B), P and Q, against beer-spoilage lactic acid bacteria in broth, wort (hopped and unhopped), and alcoholic and non-alcoholic lager beers

Enterococcus faecium L50 produces enterocin L50 (L50A and L50B) (EntL50, EntL50A and EntL50B), enterocin P (EntP) and enterocin Q (EntQ) and displays a broad antimicrobial spectrum against the most relevant beer-spoilage lactic acid bacteria (LAB) (i.e., Lactobacillus brevis and Pediococcus damnosus), which is mainly due to the production of EntL50 (EntL50A and EntL50B). Bacteriocin assays using in vitro-synthesized EntL50 (EntL50A and EntL50B) showed that both individual peptides possess antimicrobial activity on their own, EntL50A being the most active, but when the two peptides were combined a synergistic effect was observed. The only virulence genes detected in E. faecium L50 were efaAfm (cell wall adhesin) and ccf (sex pheromone), and this strain was susceptible to most clinically relevant antibiotics. E. faecium L50 survived but did not grow nor showed antimicrobial activity in hopped and unhopped wort, and alcoholic (1 and 5% ethanol, v/v) and non-alcoholic (0% ethanol, v/v) commercial lager beers. However, when unhopped wort was supplemented with 50% (v/v) MRS broth, E. faecium L50 grew and exerted antimicrobial activity similarly as in MRS broth. The enterocins produced by this strain were bactericidal (5 log decrease) against P. damnosus and Lb. brevis in a dose- and substrate-dependent manner when challenged in MRS broth, wort (hopped and unhopped), and alcoholic (1 and 5% ethanol, v/v) and non-alcoholic (0% ethanol, v/v) lager beers at 32 degrees C, and no bacterial resistances were detected even after incubation for 6-15 days. The enterocins in wort and lager beer (5% ethanol, v/v) withstood the heat treatments commonly employed in the brewing industry during mashing, wort boiling, fermentation, and pasteurization, and retained most of their antimicrobial activity in lager beer (5% ethanol, v/v) after long-term storage at 8 and 25 degrees C.     

Effect of overexpression of LAS17 on stress tolerance and the stability of extrachromosomal DNA in Saccharomyces cerevisiae

The effects of the overexpression of LAS17/BEE1, which encodes a yeast protein exhibiting sequence homology to the Wiscott-Aldrich syndrome protein, on the cell growth of Saccharomyces cerevisiae were examined. Sake yeast strain UT-1 grows at a faster rate as a result of the overexpression of LAS17 than control cultures under various stresses such as high temperature, high ethanol concentration, and oxidative stress, and the tolerance to these stresses was increased compared with the control. Moreover, a high cell survival rate was attained with overexpression of LAS17, when cells in the stationary phase of the growth cycle were subjected to heat killing (48 degrees C) or ethanol killing (20% v/v). In addition, the rate of induction of rho- was markedly reduced by overexpression of LAS17 when serine, tyrosine, and aspartic acid were used as N sources and the yeast was cultured at 35 degrees C, while rho- strains in control cultures were induced at a high frequency. After the incubation of cells harboring a multicopy vector in YPD or synthetic complete medium, almost all of the cells inherited the vector at about 15 copies per cell as a result of the overexpression of LAS17, whereas the cells harboring the control vector accounted for only 15% of the total number of cells. These results suggest that Las17p might be a multifunctional protein involved in cell growth regulation, extrachromosomal DNA transportation and stress responses.     

Tea extraction methods in relation to control of epimerization of tea catechins

Effect of water temperature and ethanol concentration on epimerization and extractability of tea catechins was investigated. The results showed that epigallocatechin gallate (EGCG) and epicatechin gallate (ECG) were partially epimerized into gallocatechin gallate (GCG) and catechin gallate (CG), respectively, when tea catechins extract was heated in water solution at 100 °C for 2 h or dry tea was extracted in water at 100 °C. The epimerization of the catechins was inhibited if the tea catechins extract was heated as solid powder and the dry tea was extracted in 50% (v/v) ethanol or in water at 80 °C or below. When the dry tea was extracted in water, the extractability of catechins increased with the increase of extraction temperature up to 100 °C, but there was no statistically significant difference in total catechins between 80 °C and 100 °C. When teas were extracted using ethanol solutions, the highest extractability of total catechins was obtained in 50% (v/v) ethanol for dry tea and in 75% (v/v) ethanol for fresh tea leaf. In order to reveal the real profiles of tea catechins in teas to be tested, dry tea should be extracted in 50% (v/v) ethanol for 10 min, while fresh tea leaf should be extracted in 75% (v/v) ethanol for 10 min. For commercial extraction, temperature should be controlled at 80 °C if water is used as the solvent. Copyright © 2007 Society of Chemical Industry     

Heat shock induces thermotolerance and inhibition of lysis in a lysogenic strain of Lactococcus lactis.

In this preliminary work, the heat shock response of lactic acid bacteria was investigated and characterized. Log-phase Lactococcus lactis cells pre-incubated at 40 degrees C before heat challenge at 52 degrees C for 30 min demonstrated increased thermotolerance as compared with cells pre-incubated at 30 degrees C. The response persisted for at least 60 min. Additionally, we demonstrated that: (i) the physiological expression of the heat shock response is temperature dependent; (ii) ethanol 4.0% (v/v) caused, to a lesser extent, a response similar to the heat shock; and (iii) hydrogen peroxide failed to induce a detectable response. Furthermore, we suggest that the induction of the heat shock response increases the resistance of a lysogenic strain of L. lactis, treated by mitomycin C (1.25 micrograms/ml), to lysis by the bacteriophage.     

Isolation of a novel mutant strain of Saccharomyces cerevisiae by an ethyl methane sulfonate-induced mutagenesis approach as a high producer of bioethanol

In order to obtain mutant strains showing higher bioethanol production than wild-type strains, a commercial Saccharomyces cerevisiae type was subjected to mutagenesis using ethyl methane sulfonate (EMS). After adding EMS to a shaken yeast suspension, the viability of yeast cells was assessed by diluted sample inoculation to solid yeast-extract peptone glucose (YEPG) medium at 15-min intervals. At 45 min, the viability of yeast cells was estimated to be about 40%. Mutagenized cells were recovered from YEPG broth after incubation at 30 degrees C for 18 h. After this period, EMS-treated yeast cells were grown on solid aerobic low-peptone (ALP) medium containing 2-12% (v/v) ethanol. All plates were incubated at 30 degrees C for 2-6 d in order to form colonies. The mutant strains that tolerated high concentrations of ethanol were selected for bioethanol production in microfuge tubes containing fermentation medium. Formation of bioethanol in small tubes was detected by the distillation-colorimetric method. In addition, trehalose content and invertase activity were determined in each mutant strain. Among many isolated mutant strains, there were six isolated colonies that grew on ALP medium supplemented with 10% (v/v) ethanol and one of them produced bioethanol 17.3% more than the wild type.     

Influence of food system conditions on N-acyl-L-homoserine lactones production by Aeromonas spp

Eleven of 13 Aeromonas strains were shown to produce AHLs. Results of TLC showed that N-butanoyl-L-homoserine lactone (C4-HSL) was the main AHL produced in LB medium at 30 degrees C. The influence of different carbon sources, temperature, pH values and salt concentrations on AHL production was determined in eight A. hydrophila and one A. caviae strain. Additionally a quantitative study of C4-HSL production by A. hydrophila strain 519 under different conditions was performed. Positive results were found in the AHL induction assay for some Aeromonas strains in cultures in LB agar incubated at 12 degrees C after 72-96 h. The induction of the sensor strains by Aeromonas spp. occurred in LB medium supplemented with all carbon sources in a concentration of 0.5%. The production of C4-HSL by A. hydrophila 519 was found until 3.5% (w/v) of NaCl. For pHs close to the neutrality the C4-HSL production by A. hydrophila was evident after 24-48 h of incubation. A. hydrophila 519 produced C4-HSL under anaerobic conditions. Also, the AHL production by Aeromonas strains was studied in simulate agar of shrimp, fish and some vegetables. The production of AHLs was evident by almost all the test strains in shrimp simulated agar. In fish agar only for one of three fish species tested, positive results were found. Induction assay in vegetables simulated agar showed principally negative results, probably because of the presence of inhibitory compounds in these vegetables.     

Survival of Vibrio parahaemolyticus under environmental stresses as influenced by growth phase and pre-adaptation treatment

In this study, the susceptibility of Vibrio parahaemolyticus in different growth phases after exposure to lethal stresses including 47 °C and 8% ethanol was first investigated. The effect of a culture's growth phase on both the heat and ethanol shock response of V. parahaemolyticus was then examined. It was found that cells of V. parahaemolyticus in the mid-exponential phase, regardless of adaptation, were most susceptible to environmental stresses, while cells in the stationary phase were least susceptible to the lethal stresses examined. Adaptation with heat shock at 42 °C for 45 min or ethanol shock with 5% ethanol for 60 min induced an increased resistance of V. parahaemolyticus to subsequent lethal stresses at 47 °C and 8% ethanol. While the adaptation treatments resulted in a reduced resistance of the test organism to pH 4.4 and 20% NaCl. Generally, the extent of changes in the resistance of V. parahaemolyticus to lethal stresses between the adapted and control cells was found to be growth phase dependent. Compared with the respective control cells, the adapted late-exponential phase cells exhibited the greatest extent of change, while the adapted stationary phase cells showed the least change in their resistance to the lethal stresses examined.     

Temperature shift as a process optimization step for the production of pro-urokinase by a recombinant Chinese hamster ovary cell line in high-density perfusion culture

Based on the effects of temperature shift on the cell cycle, apoptosis and metabolism of a recombinant Chinese hamster ovary (rCHO) cell line (CL-11G) producing pro-urokinase (pro-UK) in batch cultures, the potential of temperature shift as a tool in the optimization of the perfusion culture of CL-11G cells for the production of pro-UK was examined. The proportion of CL-11G cells in the G0/G1 phase in static cultures increased from 56.4% to 82.8% following a temperature shift from 37 degrees C to 31 degrees C. Conversely, the proportion of CL-11G cells in the S phase decreased from 34.8% to 11.6%. The specific growth rate of CL-11G cells reflected the effect of temperature on the cell cycle and decreased from 0.024 h(-1) at 37 degrees C to 0.006 h(-1) at 31 degrees C. Continuous exposure to the non-permissive temperature of 31 degrees C led to a marginal increase in apoptosis. The specific pro-UK productivity of CL-11G cells increased by 74% at 34 degrees C compared with controls at 37 degrees C in batch cultures. CL-11G cells immobilized with Cytopore 1 in a 5-l bioreactor initiated at 37 degrees C and temperature shifted to 34 degrees C exhibited an average 17% increase in viable cell density and an average 47% increase in pro-UK production. These results demonstrated that temperature shift offers the prospect of enhancing the productivity of pro-UK in high-density perfusion culture.     

Antibacterial effect of protamine in combination with EDTA and refrigeration.

The antimicrobial effect of protamine (clupeine) on a range of gram-positive and gram-negative foodborne pathogens and spoilage bacteria, was evaluated using an agar dilution assay and a broth dilution assay with Alamar Blue as growth indicator. Protamine was tested alone at concentrations from 0 to 10,000 microg/ml, and in combination with EDTA (0.9 mM). Assays were performed at 5 degrees C, 10 degrees C, 18 degrees C and 30 degrees C to test the effect of temperature. Minimum inhibitory concentration (MIC) values ranged from 10 microg/ml for Brochothrix thermosphacta to no inhibition at 10,000 microg/ml for bacteria such as Aeromonas hydrophila, proteolytic strains of Clostridium botulinum, Hafnia alvei and Morganella morganii. The minimum bactericidal concentrations (MBCs) were generally higher than MICs. In combination with EDTA, MICs of protamine decreased for gram-negative test strains, whereas EDTA alone inhibited gram-positive strains. The effect of assay incubation temperature was variable and not clear for most strains. Concentrations of 100-750 microg/ml protamine inhibited the five non-proteolytic C. botulinum strains, while none of the eight proteolytic strains was inhibited, indicating the possible role of proteolytic enzymes in protecting cells from protamine. Clearing zones, indicative of proteolytic activity, were observed in the opaque TSB-agarose around colonies of some but not all protamine-resistant bacteria, suggesting that this is not the only resistance mechanism. Addition of 5% (w/v) gelatin to study the effect of an increased protein concentration in the agar dilution assay showed that electrostatic interactions between protamine and the protein decreased the antimicrobial efficacy of the peptide.     

Determination of viability of Aeromonas hydrophila in increasing concentrations of sodium chloride at different temperatures by flow cytometry and plate count technique

Aeromonads in waters and foods can represent a risk to human health. Factors such as sodium chloride concentration and temperature can affect growth and viability of several food and water-borne pathogens. The behaviour of an Aeromonas hydrophila strain in the presence of 1.7%, 3.4% and 6% NaCl concentrations at 24 degrees C and 4 degrees C was studied over a 188 day period. Viability and membrane potential were assessed by flow cytometry; growth was evaluated by plate count technique. Flow cytometry evidenced that A. hydrophila retained viability over the period although varying according to temperature and salt concentrations. Colony Forming Units were generally lower in number than viable cells especially in the presence of 6% NaCl, indicating the occurrence of stressed cells which maintain metabolic activity yet are not able to grow on agar plates. In conclusion, A. hydrophila showed a long-term halotolerance even at elevated (6%) NaCl concentrations and a lesser sensitivity to salt at low temperature; therefore, low temperature and salt, which are two important factors limiting bacterial growth, do not assure safety in the case of high initial contamination. Finally, cytometry appears a valid tool for the rapid detection of the viability of pathogenic bacteria in food and environmental matrices to control and prevent health risks.     

Selection of entomopathogenic fungi for aphid control

Twelve strains of entomopathogenic fungi such as Lecanicillium lecanii, Paecilomyces farinosus, Beauveria bassiana, Metarhizium anisopliae, Cordyceps scarabaeicola, and Nomuraea rileyi were screened for aphid control. At 25 degrees C and 75% relative humidity (RH), among tested entomopathogenic fungi, L. lecanii 41185 showed the highest virulent pathogenicity for both Myzus persicae and Aphis gossypii, and their control values were both nearly 100% 5 and 2 d after treatment, respectively. Moreover, at an RH of 45% and in a wide temperature range (20-30 degrees C), L. lecanii 41185 also exhibited the highest virulence to M. persicae. The control value of M. persicae and the 50% lethal time (LT50) decreased significantly as the applied conidial concentration increased. The 50% lethal concentration (LC50) of the conidial suspension of this fungus was determined to be 6.55x10(5) conidia/ml. The control values of M. persicae resulting from the application of 1x10(7) and 1x10(8) conidia/ml were nearly the same and were significantly higher than that of 1x10(6) conidia/ml. The tested entomopathogenic fungi grew in a broad temperature range (15-30 degrees C). Lecanicillium strains showed optimum growth at 25 degrees C. The aerial conidia of Lecanicillium strains also could germinate in a broad temperature range (15-30 degrees C) and L. lecanii 41185 was the only strain with conidial germination at 35 degrees C.     

Growth of Aeromonas hydrophila in the whey cheeses Myzithra, Anthotyros, and Manouri during storage at 4 and 12 degrees C

The fresh whey cheeses Myzithra, Anthotyros, and Manouri were inoculated with Aeromonas hydrophila strain NTCC 8049 (type strain) or with an A. hydrophila strain isolated from food (food isolate) at levels of 3.0 to 5.0 x 10(2) CFU/g of cheese and stored at 4 or 12 degrees C. Duplicate samples of cheeses were tested for levels of A. hydrophila and pH after up to 29 days of storage. At 4 degrees C, A. hydrophila grew in Myzithra and Anthotyros with a generation time of ca. 19 h, but no growth was observed in Manouri. In Myzithra, average maximum populations of 8.87 log CFU/g (type strain) and 8.79 log CFU/g (food isolate) were recorded after 20 and 22 days of storage at 4 degrees C, respectively. The average maximum populations observed in Anthotyros stored at 4 degrees C were 6.72 log CFU/g (food isolate) and 6.13 log CFU/g (type strain) and were observed after 15 and 16 days of storage, respectively. A. hydrophila grew rapidly and reached high numbers in cheeses stored at 12 degrees C. The average generation times were 3.7 and 3.9 h (Myzithra), 4.1 and 6.1 h (Anthotyros), and 8.0 and 9.2 h (Manouri) for the type strain and the food isolate, respectively. Among the different whey cheese trials, the highest A. hydrophila population recorded (10.13 log CFU/g) was in Myzithra that had been inoculated with the food isolate after 8 days of storage at 12 degrees C. To prevent A. hydrophila growth in whey cheeses, efforts must be focused on preventing postprocessing contamination and temperature abuse during transportation and storage.     

Behavior of aeromonas hydrophila in bottled mineral waters.

The growth and survival of Aeromonas hydrophila in three types of natural mineral waters were investigated. Mineral waters with different levels of mineral content (low, medium, and high) were experimentally contaminated with A. hydrophila, stored at different temperatures (10 degrees C and 20 degrees C), and analyzed at intervals over a 60-day period. Water samples that were not experimentally contaminated were investigated for indigenous A. hydrophila. The results confirmed that A. hydrophila may occur naturally in mineral waters and showed that the level of mineral content, temperature, length of storage, and, in some cases, the type of container used may favor the growth of A. hydrophila. The greatest proliferation was observed in water with a low mineral content stored in PET bottles at 10 degrees C, in which A. hydrophila peaked at day 28 (4.47 +/- 0.01 log CFU/100 ml). At 20 degrees C, the same load was observed at day 60. The presence of high densities of A. hydrophila in bottled mineral water can constitute a risk for some groups of consumers, such as elderly and immunocompromised persons.     

Increase of the ATP-dependent phosphoenolpyruvate carboxykinase activity in Sinorhizobium meliloti (Rhizobium meliloti) during hypothermic environmental conditions

Sinorhizobium meliloti growth is affected when the incubation temperature is lower than 22 degrees C. In culture media containing glucose or fructose (1%, w/v), the doubling time at 19 degrees C was about 6.25 h during the exponential growth phase, while it was 2.75 h at 30 degrees C; at 17 degrees C it was three-fold higher than at 30 degrees C. Modifications in the bacterial metabolism explain the doubling time increase when bacteria are incubated at low temperature. We determine here, the phosphoenolpyruvate carboxykinase (PEPCK) activity increases when S. meliloti cells first grown at 30 degrees C are shifted at 17 degrees C and incubated for 10 h at this low temperature; we noted the PEPCK activity was three-fold higher in cells incubated in media containing glucose and shifted from 30 to 17 degrees C than in cells maintained at 30 degrees C, while it was only 1.5-fold higher in cells grown in media containing fructose.     

Hemolytic and proteolytic activities of Aeromonas hydrophila and Aeromonas veronii biovar sobria in broth and salmon extract at different temperatures

Expression of hemolytic and proteolytic activities throughout the growth cycle was investigated with two enterotoxic aeromonad strains assigned to the species Aeromonas hydrophila and Aeromonas veronii biovar sobria. Although growth kinetic data were dependent on strain, temperature, and substrate, maximum populations attained were higher than 9 log CFU/ml in aerated tryptone soya broth plus yeast extract (TSBYE) and salmon extract within the range 4 to 28 degrees C. For both strains in TSBYE, variable amounts of hemolytic activity were first detected at any temperature when aeromonad counts were over 9 log CFU/ml. Afterwards, this activity increased up to similar levels (109 to 112 hemolytic units per ml) without a significant increase in populations. Salmon extract supported hemolysin synthesis at 28 but not 4 degrees C. Proteolytic activity of the A. hydrophila strain was only expressed in salmon extract at 28 degrees C, whereas A. veronii biovar sobria did at 28 degrees C in both substrates and at 10 degrees C in TSBYE.     

Short communication: Salt tolerance of Lactococcus lactis R-604 as influenced by mild stresses from ethanol,heat, hydrogen peroxide,and UV light

Lactococcus lactis is a culture widely used in salt-containing dairy products. Salt hinders bacterial growth, but exposure to environmental stress may protect cells against subsequent stress, including salt. The objective of this study was to evaluate the salt tolerance of L. lactis R-604 after exposure to various stresses. The culture was subjected to 10% (vol/vol) ethanol for 30 min, mild heat at 52°C for 30 min, 15 mM hydrogen peroxide for 30 min, or UV light (254 nm) for 5 min and compared with a control. Starting with 5 log cfu/mL for all treatments, growth was determined in M17 broth with 5 NaCl concentrations (0, 1, 3, 5, and 7% wt/vol). Plating was conducted daily for 5 d. Salt tolerance was enhanced with mild heat exposure before growth in M17 broth with 5% (wt/vol) NaCl on d 3, 4, and 5, and with exposure to hydrogen peroxide and ethanol stresses before growth in M17 broth with 5% (wt/vol) NaCl on d 4 and 5. Exposure of this culture to mild heat, hydrogen peroxide, or ethanol before growth in M17 broth containing 5% (wt/vol) salt can enhance its survival, which could be beneficial when using it in salt-containing dairy products.     

Bioactive components and antioxidant properties of γ-aminobutyric acid (GABA) tea leaves

Various ethanolic concentrations (0–95%, v/v) and temperatures (25–95 °C) were used to extract γ-aminobutyric acid (GABA) tea leaves. Extraction yields, and contents of total phenols, various catechins, GABA, theanine, and antioxidant properties of extracts were determined. The 50% (v/v) ethanol at 50–95 °C gave higher yields (32.05–32.56 g dried extract/100 g dried tea leaves). The bioactive components and antioxidant properties of extracts were affected by the ethanolic concentrations and temperatures. Among catechins, epigallocatechin gallate was the main catechin in all extracts, followed by epigallocatechin, epicatechin, epicatechin gallate, gallocatechin and gallocatechin gallate. The 50–75% (v/v) ethanol at 75–95 °C gave higher contents of ester type (102.92–104.54 mg/g extract) and non-ester type (61.75–63.55 mg/g extract) catechins. Water at 50–75 °C gave higher GABA and theanine contents and higher chelating ability of extracts. The 75% (v/v) ethanol at 25–75 °C gave higher scavenging ability and reducing power of extracts. Based on dried tea extracts or leaves results obtained, the optimal extraction conditions to maintain the total contents of various catechins, GABA and theanine in the maximum level were 50% ethanol (v/v) and 75–95 °C.     

Production of a high concentration of ethanol from potato tuber by high gravity fermentation

To produce a high concentration of ethanol from viscous potato tuber mash, potato tuber mash containing high contents of solids (28%) was prepared by grinding the potato tuber without the addition of water. The viscosity of the potato mash was reduced by using Viscozyme (0.1%) at 50°C for 30 min. The potato mash was then liquefied using Liquozyme (0.1%) at 90°C for 30 min and optimal conditions for the simultaneous saccharification and fermentation (SSF) of the potato mash for ethanol production were investigated using statistical methods. Using 2⁴ factorial design, saccharifying-enzyme and incubation temperature were found to be important factors. Using response surface methodology, the optimal saccharifying-enzyme dosage and incubation temperature were determined to be 1.45 AGU/g dry matter and 31.3°C, respectively. Under these optimal conditions for SSF, 14.92%(v/v) ethanol with 91.0% of theoretical yield was produced after 60 h, and all the starch was completely used up.     

Response of Vibrio parahaemolyticus to ethanol shock

In this study, the growth and survival of Vibrio parahaemolyticus in the presence of 0.0-8.0% ethanol was first examined. V. parahaemolyticus was then exposed to a sub-lethal dose of 5.0% ethanol for 30 and 60 min (ethanol shock). Morphological changes and alterations in cell leakage, thermal tolerance at 47 degrees C, and susceptibility to 8% ethanol and low temperature (4 and -18 degrees C) of V. parahaemolyticus caused by ethanol shock were investigated. In addition, recoveries of the ethanol-shocked cells of V. parahaemolyticus on thiosulfate-citrate-bile salts-sucrose agar (TCBS) and TSA-3.0% NaCl were also compared. The findings revealed that the presence of ethanol in TSB-3.0% NaCl at 6.0-8.0% and 5.0% or less, exerted bactericidal and partial growth inhibition effect, respectively, on V. parahaemolyticus. Recovery of ethanol-shocked cells of V. parahaemolyticus was significantly (P<0.05) less on TCBS than on TSA-3.0% NaCl. A significantly (P<0.05) marked increase of protein and nucleic acid material in the supernatant of cell suspension was found after cells of V. parahaemolyticus were exposed to ethanol shock. Extensive cell disruption, wrinkling and cell-wall pitting, indicative of cell-surface damage were also noted on the ethanol-shocked cells. Ethanol-shocked cells of V. parahaemolyticus exhibited a similar yet higher susceptibility at 4 and -18 degrees C compared with the control cells. Moreover, there was a marked increase in the thermal tolerance and resistance to 8.0% ethanol with cells of V. parahaemolyticus after ethanol shock. Finally, the duration of ethanol shock testing did not affect the extent of increased thermal tolerance. While cells of V. parahaemolyticus subjected to ethanol shock for 60 min showed an increase in their resistance to 8.0% ethanol, they also showed an increase in susceptibility at -18 degrees C, than those ethanol shocked for 30 min.     

Effect of packaging atmosphere on the microbial attributes of pearlspot (Etroplus suratensis Bloch) stored at 0-2 degrees C

Effect of packaging atmosphere (air and under different modified atmospheres (MAs), 40% CO2/60% O2, 50%/50% O2, 60% CO2/40% O2, 70% CO2/30% O2 and 40% CO2/30% O2/30% N2) on the microbial and biochemical attributes of fresh pearlspot (Etroplus suratensis Bloch) stored at 0-2 degrees C was investigated. Trimethylamine nitrogen (TMA-N) and thiobarbituric acid (TBA) values remained lower than the proposed acceptability limits throughout the storage period. Results demonstrated that storage of pearlspot under air and MA 40% CO2/30% O(2)/30% N(2) resulted in growth of Enterobacteriaceae, Aeromonas and H(2)S-producing bacteria including Shewanella putrefaciens, while all other packaging atmospheres did not allow multiplication of Enterobacteriaceae and Aeromonas within 3 weeks. Aeromonas spp. identified were Aeromonas hydrophila, Aeromonas veronii biovar sobria and A. veronii biovar veronii. Significant reduction (p<0.01) was noticed in Aeromonas population of pearlspot stored under MA 60% CO2/40% O2 and 70% CO2/30% O2. A delay of growth of Pseudomonas below 5.0log(10)cfug(-1) was observed during the 15th day of storage at 0-2 degrees C under modified atmosphere packaging (MAP) conditions. Growth of faecal streptococci was significantly inhibited in all the packaging atmospheres at 0-2 degrees C during the entire storage period. Survival of coagulase positive Staphylococci (<50cfug(-1)) in low numbers was noticed during storage in all the packaging atmospheres. Clostridium botulinum toxin was not detected. All the packaging atmospheres did not allow multiplication of sulphite-reducing clostridia at 0-2 degrees C during the entire storage period. Packaging in MA 60% CO2/40% O2 resulted in the inhibition of growth of Aeromonas and Enterobacteriaceae, and the slowest growth of psychrotrophic bacteria, H(2)S-producing bacteria, including Shewanella putrefaciens and Pseudomonas and extended microbiological shelf life to 9-10 days. This study confirms the survival of potentially pathogenic A. hydrophila, A. veronii biovar sobria and A. veronii biovar veronii capable of growth at low temperature in pearlspot stored under MA.