Morphological changes of temperature- and pH-stressed Salmonella following exposure to cetylpyridinium chloride and nisin

The outer membrane of Gram-negative bacteria such as Salmonella, act as a permeability barrier, preventing nisin gaining access to the cytoplasmic membrane. If the outer membrane permeability is reduced, however, Gram-negative bacteria can show nisin sensitivity. In this study, temperature stresses (heating, chilling and freezing), pH stress (pH 4.5, 5.0, 6.0) and cetylpyridinium chloride (CPC)-nisin treatment were used to alter the outer membrane permeability of Salmonella, producing a loss of barrier function and reduced resistance to nisin. The morphological changes in Salmonella were examined using scanning electron microscopy. Temperature and pH-stressed S. typhimurium cells, untreated and treated with CPC-nisin had perturbed cell morphology, including apparent indentations and craters in the cell surfaces and collapsed amorphous bodies.     

Potential of bacteriocinogenic Lactococcus lactis subsp. lactis inhabiting low pH vegetables to produce nisin variants

Antimicrobial behavior of lactic acid bacteria (LAB) has been explored since many years to assess their ability to produce bacteriocin, a natural preservative, to increase the shelf life of food. This study aims to characterize bacteriocin producing strains of lactic acid bacteria isolated from acidic to slightly acidic raw vegetables including tomato, bell pepper and green chili and to investigate their potential to inhibit food related bacteria. Among twenty nine LAB screened for antimicrobial activity, three exhibited antagonism against closely related bacterial isolates which was influenced by varying temperature and pH. They were identified up to strain level as Lactococcus lactis subsp. lactis TI-4, L. lactis subsp. lactis CE-2 and L. lactis subsp. lactis PI-2 based on 16S rRNA gene sequence. Their spectrum of inhibition was observed against food associated strains of Bacillus subtilis and Staphylococcus aureus. Moreover, L. lactis subsp. lactis PI-2 selected on the basis of higher antimicrobial activity was further evaluated for bacteriocin production which was detected as nisin A and nisin Z. These findings suggest the possible use of L. lactis strains of vegetable origin as protective cultures in slightly acidic as well as slightly alkaline food by the bio-preservative action of bacteriocins.     

Effect of nanovesicle-encapsulated nisin on growth of Listeria monocytogenes in milk

Commercial nisin was encapsulated in nanovesicles (mean diameter 140 nm) prepared from partially purified soy lecithin. Nisin-loaded liposomes and unencapsulated (free) nisin were initially tested in BHI medium and skim milk inoculated with Listeria monocytogenes and incubated for 48 h at 30 °C. At such abuse temperature conditions, free nisin showed better inhibitory than the liposomal counterparts. Subsequently, the effect of encapsulated or free nisin was evaluated in combination with refrigeration (7 ± 1 °C) in both whole (3.25% fat) and skim (0% fat) milk for up to 14 day. A decrease of 3–4 log cycles in L. monocytogenes counts was observed for free and encapsulated nisin at 0.5 mg/ml concentration. Liposome encapsulation of antimicrobial peptides may be important to overcome stability issues and interaction with food components. The utilization of nanovesicle-encapsulated nisin in combination with low temperatures appeared to be effective to control L. monocytogenes in milk, emphasizing the importance of hurdle technology to assure food safety.     

Polysulfide reduction by Clostridium relatives isolated from sulfate-reducing enrichment cultures

Sulfur is almost insoluble in water at ambient temperatures, and therefore polysulfide (S_n²⁻) has been considered as a possible intermediate that is used directly by bacteria in sulfur respiration. Sulfur-reducing reductases have been purified and characterized from a few sulfur reducers. However, polysulfide reduction has only been confirmed in Wolinella succinogenes. In our previous study, the direct production of hydrogen sulfide from polysulfide was confirmed by an enrichment culture obtained from natural samples under sulfate-reducing conditions. The present study attempted to isolate and identify polysulfide-reducing bacteria from the enrichment cultures. Almost all the isolated strains were classified into the genus Clostridium, based on 16S rRNA gene sequence analysis. The isolates, and some closely related strains, were able to reduce polysulfide to hydrogen sulfide. During production of 1 mol of hydrogen sulfide, approximately 2 mol of lactate was converted to acetate. Thus, dissimilatory polysulfide reduction occurred using lactate as an electron donor. The ability to reduce elemental sulfur was also examined with the isolates and the related strains. Although elemental sulfur reducing strains can reduce polysulfides, not all polysulfide-reducing strains can reduce elemental sulfur. These results demonstrate that the conversion of elemental sulfur to polysulfide seems to be important in the reduction process of sulfur.     

Diversity of culturable microorganisms and occurrence of Listeria monocytogenes and Salmonella spp. in Tuber aestivum and Tuber melanosporum ascocarps

The aim of this study was to investigate the total mesophilic microorganisms, Pseudomonas genus, Enterobacteriaceae family, mold and yeast counts and the presence of Listeria monocytogenes and Salmonella spp on Tuber aestivum and Tuber melanosporum ascocarps. The results confirmed that the major percentage of the microorganisms, approximately 9.0 log ufc/g, were present in the peridium, the glebas of healthy truffles being practically free of microorganisms. The predominant microbial group was the Pseudomonas averaging 8.3 and 8.4 log cfu/g on T. aestivum and T. melanosporum whole ascocarps, respectively. The Enterobacteriaceae also achieved high populations, especially in T. aestivum truffles, with 6.3 log cfu/g. Molds and yeasts never exceeded 5.0 log cfu/g. The characterization of the isolates revealed that the fluorescens pseudomonads were the most prevalent. Raoultella terrigena and Enterobacter intermedius were the dominant Enterobacteriaceae. The identification of the yeast isolates revealed five species: Debaryomyces hansenii, Issatchenkia scutulata, Rhodotorula aurantiaca, Saccharomyces dairensis and Trichosporon beigelii subspecies A and B. The mold genera detected in both species of truffles were Aspergillus, Cladosporium, Penicillium and Fusarium, Trichoderma being present only in T. aestivum. L. monocytogenes was found in 10% of the samples of T. aestivum analysed but Salmonella spp. was not detected. Knowledge of the microbial population in terms of possible food borne and pathogen microorganisms is very useful for establishing successful disinfection and storage methods to prolong the shelf-life of ascocarps of T. aestivum and T. melanosporum.     

Effect of exopolysaccharides (EPSs) produced by Lactobacillus delbrueckii subsp. bulgaricus strains to bacteriophage and nisin sensitivity of the bacteria

Lactobacillus strains used in this study were isolated from village-type yogurt and raw milk. The isolates were identified as Lactobacillus delbrueckii subsp. bulgaricus by 16 s rDNA sequence analysis and API 50 CHL identification systems. The exopolysaccharide (EPS) production of the strains growth in skim milk were investigated. In addition sensitivity and insensitivity of these strains against domestic bacteriophages and nisin were examined. It was deduced that those strains which had relatively high EPS-producing capacity were insensitive against phages and nisin. Linear relationships were determined between EPS production of the bacteria and bacteriophage and nisin insensitivity of the bacteria.There was a negative correlation between EPS production quantity and phage and nisin sensitivity of the bacteria. Of all the strains, L. delbrueckii subsp bulgaricus B3 produced the highest EPS quantity, and it was insensitive against phages and nisin. Based on these results, it is suggested that L. delbrueckii subsp bulgaricus B3 can be used with the starter culture in dairy industry for stable and high-quality yogurt production.     

Response of Bacillus cereus spores to high hydrostatic pressure and moderate heat

The effects of high pressure (400-600 MPa) and moderate heat (60-80 °C) treatments at various process times (10-20 min) on the reduction of Bacillus cereus As 1.1846 spores, suspended in milk buffer were investigated. In the present work, response surface methodology (RSM) was employed, and a quadratic equation of high hydrostatic pressure inactivation was built with RSM. By analyzing response surface plots and corresponding contour plots and by solving the quadratic equation, experimental values were shown to be significantly in agreement with predicted values, since the adjusted determination coefficient was 0.9752 and the level of significance was P < 0.0001. Optimum process parameters for a six-log cycle reduction of B. cereus spores were obtained: pressure, 540.0 MPa; temperature, 71 °C; and pressure-holding time, 16.8 min. The adequacy of the model equation in predicting optimum response values was verified effectively using experimental test data that was not used in the development of the model.     

Inactivation of Vibrio parahaemolyticus and Vibrio vulnificus in oysters by high-hydrostatic pressure and mild heat

Several recent outbreaks associated with oysters have heightened safety concerns of raw shellfish consumptions, with the majority being attributed to Vibrio spp. The objective of this study was to determine the effect of high-hydrostatic pressure (HHP) followed by mild heating on the inactivation of Vibrio parahaemolyticus and Vibrio vulnificus in live oysters. Inoculated oysters were randomly subjected to: a) pressurization at 200–300 MPa for 2 min at 21 °C, b) mild heat treatment at 40, 45 or 50 °C for up to 20 min and c) pressure treatment of 200–300 MPa for 2 min at 21 °C followed by heat treatment at 40–50 °C. Counts of V. parahaemolyticus and V. vulnificus were then determined using the most probable number (MPN) method. Pressurization at 200–300 MPa for 2 min resulted in various degrees of inactivation, from 1.2 to >7 log MPN/g reductions. Heat treatment at 40 and 45 °C for 20 min only reduced V. parahaemolyticus and V. vulnificus by 0.7–2.5 log MPN/g while at 50 °C for 15 min achieved >7 log MPN/g reduction. HHP and mild heat had synergistic effects. Combinations such as HHP at 250 MPa for 2 min followed by heat treatment at 45 °C for 15 min and HHP at 200 MPa for 2 min followed by heat treatment at 50 °C for 5 min reduced both V. parahaemolyticus and V. vulnificus to non-detectable levels by the MPN method (<3 MPN/g). HHP at ≥275 MPa for 2 min followed by heat treatment at 45 °C for 20 min and HHP at ≥200 MPa for 2 min followed by heat treatment at 50 °C for 15 min completely eliminated both pathogens in oysters (negative enrichment results). This study demonstrated the efficiency of HHP followed by mild heat treatments on inactivation of V. parahaemolyticus and V. vulnificus and could help the industry to establish parameters for processing oysters.     

Membrane permeabilization and cellular death of Escherichia coli, Listeria monocytogenes and Saccharomyces cerevisiae as induced by high pressure carbon dioxide treatment

In this study, the relationship between (irreversible) membrane permeabilization and loss of viability in Escherichia coli, Listeria monocytogenes and Saccharomyces cerevisiae cells subjected to high pressure carbon dioxide (HPCD) treatment at different process conditions including temperature (35–45 °C), pressure (10.5–21.0 MPa) and treatment time (0–60 min) was examined. Loss of membrane integrity was measured as increased uptake of the fluorescent dye propidium iodide (PI) with spectrofluorometry, while cell inactivation was determined by viable cell count. Uptake of PI by all three strains indicated that membrane damage is involved in the mechanism of HPCD inactivation of vegetative cells. The extent of membrane permeabilization and cellular death increased with the severity of the HPCD treatment. The resistance of the three tested organisms to HPCD treatment changed as a function of treatment time, leading to significant tailing in the survival curves, and was dependent on pressure and temperature. The results in this study also indicated a HPCD-induced damage on nucleic acids during cell inactivation. Transmission electron microscopy showed that HPCD treatment had a profound effect on the intracellular organization of the micro-organisms and influenced the permeability of the bacterial cells by introducing pores in the cell wall.     

Effects of simulated digestion in vitro on cell wall polysaccharides from kiwifruit (Actinidia spp.)

Cell wall polysaccharides are resistant to digestion and absorption in the human small intestine and are considered to be delivered to the colon in a chemically unaltered state. In this paper, pulp from green and gold kiwifruit was subjected to in vitro upper-intestinal tract digestion and the chemical and physical changes to cell wall polysaccharides (dietary fibre) were investigated. Yields of insoluble fibre decreased slightly with simulated digestion while soluble fibre yields increased. Constituent sugar and glycosyl linkage analysis of the soluble and insoluble fibre fractions revealed that the chemical composition and structure of the non-starch polysaccharides remained largely unchanged. However, the degree of methylesterification of galacturonic acid residues present in the pectin-rich soluble fibre fractions of both fruit decreased with treatment; size-exclusion chromatography detected changes in the molecular weight profiles of these fractions. These changes may affect the physicochemical properties and fermentability of kiwifruit dietary fibre in the large intestine.     

Effects of combining microbial transglutaminase and high pressure processing treatments on the mechanical properties of heat-induced gels prepared from arrowtooth flounder (Atheresthes stomias)

The objective of this study was to evaluate the effect of setting conditions (25 °C for 2 h or 40 °C for 30 min) and combining of microbial transglutaminase (MTGase) and high pressure processing (HPP) on the mechanical properties of heat induced gels obtained from paste from arrowtooth flounder (Atheresthes stomias). Treatments included fish paste control without added MTGase, fish paste incubated with MTGase but not pressurized (MTGase + cooking), fish paste incubated with MTGase and pressurized at 600 MPa for 5 min (MTGase + HPP + cooking) and fish paste pressurized at 600 MPa for 5 min and incubated with MTGase (HPP + MTGase + cooking). The controls and the treated samples were then subjected to one of two thermal treatments: 90 °C for 15 min or 60 °C for 30 min before cooking at 90 °C for 15 min. Samples of fish paste heated at 60 °C before cooking could not be used to prepare gels for texture profile analysis (TPA). TPA showed that pressurization improved the mechanical properties of gels made from paste treated with MTGase and set at 25 °C. The opposite was observed for samples set at 40 °C. Setting at 40 °C appeared to induce proteolytic degradation of myofibrillar proteins.     

Lactic acid resistance of Shiga toxin-producing Escherichia coli and multidrug-resistant and susceptible Salmonella Typhimurium and Salmonella Newport in meat homogenate

This study compared lactic acid resistance of individual strains of wild-type and rifampicin-resistant non-O157 Shiga toxin-producing Escherichia coli (STEC) and of susceptible and multidrug-resistant (MDR) and/or MDR with acquired ampC gene (MDR-AmpC) Salmonella against E. coli O157:H7. After inoculation of sterile 10% beef homogenate, lactic acid was added to a target concentration of 5%. Before acid addition (control), after acid addition (within 2 s, i.e. time-0), and 2, 4, 6 and 8 min after addition of acid, aliquots were removed, neutralized, and analyzed for survivors. Of wild-type and of rifampicin-resistant non-O157 STEC strains, irrespective of serogroup, 85.7% (30 out of 35 strains) and 82.9% (29 out of 35 strains), respectively, reached the detection limit within 0–6 min. Of Salmonella strains, 87.9% (29 out of 33 isolates) reached the detection limit within 0–4 min, irrespective of antibiotic resistance phenotype. Analysis of non-log-linear microbial survivor curves indicated that non-O157 STEC serogroups and MDR and susceptible Salmonella strains required less time for 4D-reduction compared to E. coli O157:H7. Overall, for nearly all strains and time intervals, individual strains of wild-type and rifampicin-resistant non-O157 STEC and Salmonella were less (P < 0.05) acid tolerant than E. coli O157:H7.     

Combined pH and high hydrostatic pressure effects on Lactococcus starter cultures and Candida spoilage yeasts in a fermented milk test system during cold storage

The combined effects of high pressure processing (HPP) and pH on the glycolytic and proteolytic activities of Lactococcus lactis subsp. lactis, a commonly used cheese starter culture and the outgrowth of spoilage yeasts of Candida species were investigated in a fermented milk test system. To prepare the test system, L. lactis subsp. lactis C10 was grown in UHT skim milk to a final pH of 4.30 and then additional samples for treatment were prepared by dilution of fermented milk with UHT skim milk to pH levels of 5.20 and 6.50. These milk samples (pH 4.30, 5.20 and 6.50) with or without an added mixture of two yeast cultures, Candida zeylanoides and Candida lipolytica (10⁵ CFU mL⁻¹ of each species), were treated at 300 and 600 MPa (≤20 °C, 5 min) and stored at 4 °C for up to 8 weeks. Continuing acidification by starter cultures, as monitored during storage, was substantially reduced in the milk pressurised at pH 5.20 where the initial titratable acidity (TA) of 0.40% increased by only 0.05% (600 MPa) and 0.10% (300 MPa) at week 8, compared to an increase of 0.30% in untreated controls. No substantial differences were observed in pH or TA between pressure-treated and untreated milk samples at pH 4.30 or 6.50. The rate of proteolysis in milk samples at pH values of 5.20 and 6.50 during storage was significantly reduced by treatment at 600 MPa. Treatment at 600 MPa also reduced the viable counts of both Candida yeast species to below the detection limit (1 CFU mL⁻¹) at all pH levels for the entire storage period. However, samples treated at 300 MPa showed recovery of C. lipolytica from week 3 onwards, reaching 10⁶–10⁷ CFU mL⁻¹ by week 8. In contrast, C. zeylanoides did not show any recovery in any of the pressure-treated samples during storage.     

Inactivation of Salmonella and Escherichia coli O157:H7 on artificially contaminated alfalfa seeds using high hydrostatic pressure

Alfalfa sprouts contaminated with Salmonella and Escherichia coli O157:H7 have been implicated in several outbreaks of foodborne illnesses in recent years. The seed used for sprouting appears to be the primary source of pathogens. Seed decontamination prior to sprouting presents a unique challenge for the sprouting industry since cells of the pathogenic survivors although undetectable after sanitizing treatments, can potentially multiply back to hazardous levels. The focus of this study was to therefore test the efficacy of high hydrostatic pressure to eliminate a ∼5 log CFU/g load of Salmonella and E. coli O157:H7 on alfalfa seeds. Pressure treatment of 600 MPa for up to 25 min at 20 °C could not result in complete inactivation of Salmonella. High-pressure treatment was then carried out either at sub-ambient (4 °C) or elevated (40, 45 and 50 °C) temperatures to test the ability of high pressure to eliminate Salmonella. Pressure treatment at 4 and 20 °C did not deliver any satisfactory inactivation of Salmonella while high pressure at elevated temperatures achieved complete kill. Pre-soaking seeds prior to high-pressure treatment also enhanced pressure inactivation of Salmonella but at the expense of seed viability. High-pressure treatment of 500 MPa for 2 min at 45 °C was able to eliminate wild-type Salmonella and E. coli O157:H7 strains without bringing about any appreciable decrease in the seed viability.     

Inactivation of Serratia liquefaciens on dry-cured ham by high pressure processing

To quantify the inactivation of Serratia liquefaciens exerted by high pressure processing (HPP), slices of dry-cured ham were inoculated and processed combining different levels of technological parameters: pressure (347–852 MPa), time (2.3–15.8 min) and temperature (7.6–24.4 °C) according to a central composite design. Bacterial inactivation, as logarithmic reduction, indicated that S. liquefaciens was relatively sensitive to HPP. Six log reductions were achieved in a total of 10 trials combining pressures of 600 MPa or above with different holding times and temperatures. The inactivation of S. liquefaciens was analysed through the multiple regression analysis to generate a second order polynomial equation. Pressure and time were the two factors which significantly determined the inactivation of S. liquefaciens on dry-cured ham. Temperature did not significantly affect the lethality of the process. The response surface methodology was used to determine optimum process conditions to maximize the inactivation of S. liquefaciens in the experimental range tested. The maximum inactivation of S. liquefaciens in dry-cured ham was achieved by combining a pressure of 650 MPa with a holding time of 8 min. Combinations above these values (i.e. 750 MPa for 13 min) would not significantly improve the lethality of the process.     

Inactivation of spores of Bacillus cereus in cheese by high hydrostatic pressure with the addition of nisin or lysozyme

The objective of this work was to study high hydrostatic pressure (HHP) inactivation of spores of Bacillus cereus ATCC 9139 inoculated in model cheeses made of raw milk, together with the effects of the addition of nisin or lysozyme. The concentration of spores in model cheeses was approximately 6-log10 cfu/g of cheese. Cheeses were vacuum packed and stored at 8 degrees C. All samples except controls were submitted to a germination cycle of 60 MPa at 30 degrees C for 210 min, to a vegetative cells destruction cycle of 300 or 400 MPa at 30 degrees C for 15 min, or to both treatments. Bacillus cereus counts were measured 24 h and 15 d after HHP treatment. The combination of both cycles improved the efficiency of the whole treatment. When the second pressure-cycle was of 400 MPa, the highest inactivation (2.4 +/- 0.1 log10 cfu/g) was obtained with the presence of nisin (1.56 mg/L of milk), whereas lysozyme (22.4 mg/L of milk) did not increase sensitivity of the spores to HHP. For nisin (0.05 and 1.56 mg/L of milk), no significant differences were found between counts at 24 h and 15 d after treatment. Considering that mesophilic spore counts usually range from 2.6 to 3.0 log10 cfu/ml in raw milk, HHP at mild temperatures with the addition of nisin may be useful for improving safety and preservation of soft curd cheeses made from raw milk.     

In vitro antimicrobial activity of essential oil from endemic Origanum minutiflorum on ciprofloxacin-resistant Campylobacter spp.

This study evaluated the antimicrobial activities of an essential oil of Origanum minutiflorum (O. Schwarz and P.H. Davis) against ciprofloxacin-resistant Campylobacter spp., by broth microdilution and agar well-diffusion methods. Moreover, O. minutiflorum oil was analyzed by gas chromatography/mass spectrometry (GC/MS). Twenty-nine components were identified, representing 98.7 of the oil. The oil yield from the plants was 4.0–4.4% v/w. The major components of O. minutiflorum oil were carvacrol (73.9%) and p-cymene (7.20%). The oil has lower contents of carvacrol methyl ether (0.05%), heptadecanol (0.06%) and carvacryl acetate (0.06%). Twenty-one Campylobacter spp. (12 C. jejuni, 5 C. lari and 4 C. coli) strains using in this study were selected among 300 isolates according to their resistance to ciprofloxacin. The minimum inhibitory concentration (MIC) values for bacterial strains, which were sensitive to the essential oil of O. minutiflorum, were in the range of 7.8–800 μg/ml. The essential oil obtained showed strong antimicrobial activity against all of the tested ciprofloxacin-resistance Campylobacter spp. These results suggest that the essential of O. minutiflorum may be used as a natural preservative in food against food-born disease, such as Campylobacteriosis.     

Antimicrobial resistance and co-selection phenomenon in Listeria spp. recovered from food and food production environments

Antimicrobial resistance (AMR), co-selection phenomenon, and the relationship between reduced susceptibility (RSC) to ciprofloxacin (CIP) and resistance to other antimicrobials in Listeria spp. (n = 103) recovered from food processing environments (FPE) and food were investigated. Resistance of Listeria monocytogenes and other listeriae, respectively, to cefoxitin (FOX; 98% vs. 88%), CIP (7% vs. 4%), clindamycin (CLI; 33% vs. 59%) and tetracycline (6% vs. 8%) was observed, as was RSC to CIP (67% vs. 57%) and CLI (65% vs. 41%). L. monocytogenes also possessed RSC to linezolid (LZD; 6%), rifampicin (2%) and streptomycin (6%), with other listeriae displaying RSC to chloramphenicol (4%). L. monocytogenes serotype 1/2a (90%) isolates were more frequently resistant or possessed RSC to CIP compared to serotype 4b (55%) (p = 0.015). When eight strains were experimentally adapted to high concentrations of CIP, co-selection occurred as MICs to benzalkonium chloride (BAC) increased (n = 5), gentamicin MICs remained the same (n = 6) or increased 2-fold (n = 2), and led to RSC to LZD (n = 1) and resistance to CLI (n = 8). Overall, levels of resistance/RSC to CIP in food chain isolates, particularly 1/2a, are concerning. Further, reduced sensitivity to disparate antimicrobials following CIP exposure highlights the need for increased knowledge of co-selection phenomenon linked with antimicrobial agents.     

Effect of media,additives, and incubation conditions on the recovery of high pressure and heat-injured Clostridium botulinum spores

The effect of additives and post-treatment incubation conditions on the recovery of high pressure and heat-injured (i.e., processed at 620 MPa and 95 and 100 °C for 5 min) spores of Clostridium botulinum strains, 62-A (proteolytic type A) and 17-B (nonproteolytic type B) was studied. High pressure and heat-injured spores were inoculated into TPGY (Trypticase–Peptone–Glucose–Yeast extract) anaerobic broth media containing additives (lysozyme, l-alanine, l-aspartic acid, dipicolonic acid, sodium bicarbonate, and sodium lactate) at various concentrations (0–10 μg/ml) individually or in combination. The spore counts of high pressure and heat-injured 62-A and 17-B recovered from TPGY broth containing lysozyme (10 μg/ml) incubated for 4 months versus that recovered from peptone–yeast extract–glucose–starch (PYGS) plating agar containing lysozyme (10 μg/ml) incubated under anaerobic conditions for 5 days were also compared. None of the additives either individually or in combination in TPGY broth improved recovery of injured spore enumeration compared to processed controls without additives. Addition of lysozyme at concentrations of 5 and 10 μg/ml in TPGY broth improved initial recovery of injured spores of 17-B during the first 4 days of incubation but did not result in additional recovery at the end of the 4 month incubation compared to the processed control without lysozyme. Adding lysozyme at a concentration of 10 μg/ml to PYGS plating agar resulted in no effect on the recovery of high pressure and heat-injured 62-A and 17-B spores. The recovery counts of high pressure and heat-injured spores of 62-A and 17-B were lower (i.e., <1.0 log units) with PYGS plating agar compared to the MPN method using TPGY broth as the growth medium.     

Fatty acids profile of selected Artemisia spp. plants: Health promotion

In the present study we report the fatty acids profile of thirteen species of Artemisia, a hardy herb or shrub, analyzed by gas chromatography connected to a mass detector (GC-MS) for their nutritional value and their potential exploitation as a new source of essential fatty acids. Total lipids content ranged from 3.31 ± 0.19 to 17.78 ± 0.27 mg/g (fresh weight). The three most abundant fatty acids were C16:0, C18:2ω6 and C18:3ω3. Unsaturated fatty acids predominated in all the Artemisia species are studied with the α-linolenic acid (ALA) and linoleic acid (LA), which are essential for normal human growth, health promotion, and disease prevention. The predominant ω3 PUFA acid in all Artemisia species analyzed, was linolenic acid, with Artemisia gmellini, Artemisia ludoviciana and Artemisia vulgaris, showing higher amounts of this fatty acid, all thirteen species, analyzed in this study, were also rich in oleic acid (ω9) and linoleic acid (ω6), accounted for 50-70% of total PUFA. The ratio of ω3 PUFA to ω6 PUFA was similar in all species, varying from 1.0 to 3.0. Identifying Artemisia species as newer sources of PUFAs and enriching or optimizing the ω3FAs in known plant sources offer us ways of increasing the availability of ω3FAs in the food supply.