Initial screening for inhibitory activity of essential oils on growth of Fusarium verticillioides, F. proliferatum and F. graminearum on maize-based agar media

An in vitro initial screening of a range of 37 essential oils on inhibition of mycelial growth of Fusarium verticillioides, F. proliferatum and F. graminearum under different temperature (20–30°C) and water activity (a_w) (0.95–0.995) conditions was made. The basic medium was a 3% maize meal extract agar. The maize meal agar was modified with glycerol to a range of water activity conditions and the essential oils were incorporated at different concentrations (0, 500, 1000 μg ml⁻¹). Cinnamon leaf, clove, lemongrass, oregano and palmarosa oils were the products tested suitable for being used as novel preservatives in the control of the three Fusarium species studied. Although water activity was determinant for the growth of the isolates, in general, the preservative effects of the oils were not linked to a_w. However, a trend to a higher inhibition by the oils when a_w was low was observed. Temperature had a minor importance in the inhibitory effect of the preservatives. In vivo studies may be required to confirm the usefulness of the results obtained.     

Essential oil composition and antibacterial activity of Thymus caramanicus at different phenological stages

Thymus species are well known as medicinal plants because of their biological and pharmacological properties. Thymus caramanicus is an endemic species grown in Iran. Variation in the quantity and quality of the essential oil of wild population of T. caramanicus at different phenological stages including vegetative, floral budding, flowering and seed set are reported. The oils of air-dried samples were obtained by hydrodistillation. The yields of oils (w/w%) at different stages were in the order of: flowering (2.5%), floral budding (2.1%), seed set (2.0%) and vegetative (1.9%). The oils were analyzed by GC and GC–MS. In total 37, 37, 29 and 35 components were identified and quantified in vegetative, floral budding, full flowering and seed set, representing 99.3, 98.6, 99.2 and 97.8% of the oil, respectively. Carvacrol was the major compound in all samples. The ranges of major constituents were as follow: carvacrol (58.9–68.9%), p-cymene (3.0–8.9%), γ-terpinene (4.3–8.0%), thymol (2.4–6.0%) and borneol (2.3–4.0%). Antibacterial activity of the oils and their main compounds were tested against seven Gram-positive and Gram-negative bacteria by disc diffusion method and determining their minimum inhibitory concentration (MIC) values. The inhibition zones (IZ) and MIC values for bacterial strains, which were sensitive to the essential oil of T. caramanicus, were in the range of 15–36 mm and 0.5–15.0 mg/ml, respectively. The oils of various phenological stages showed high activity against all tested bacteria, of which Bacillus subtilis and Pseudomonas aeruginosa were the most sensitive and resistant strains, respectively. Thus, they represent an inexpensive source of natural antibacterial substances that exhibited potential for use in pathogenic systems.     

Supercritical fluid extraction of<Emphasis Type="Italic"> Cucurbita ficifolia</Emphasis> seed oil

Pumpkin, Cucurbita ficifolia, seed oil was extracted with supercritical carbon dioxide (SC-CO₂) in the temperature range of 308–318 K and in the pressure range of 18–20 MPa. In addition, the influence of the superficial velocity within a tubular extractor was studied. The oil content determined by a Soxhlet apparatus was 43.5%. Physical and chemical characteristics of the oil were obtained. The results in terms of free fatty acids contents were compared with those obtained when n-hexane was used as the solvent, and no significant differences between the oils extracted by both methods were found. The main fatty acid was 6-linoleic acid (about 60%), followed by palmitic acid (about 15%) and oleic acid (about 14%). Oxidative stability was studied by using the induction time determined by the Rancimat method. The oil obtained by supercritical fluid extraction (SFE) was less protected against oxidation (4.2 h for SFE-extracted oil and 8.3 h for the pumpkin seed oil extracted with n-hexane). The oil extracted by SC-CO₂ was clearer than that extracted by n-hexane, showing some refining. The acidity index was 5.5 for the n-hexane extracted oil. For the oils extracted by SC-CO₂, two analyses were made: for the oils obtained at 15 min of extraction time, for which the acidity indices varied from about 15 to 20, and for the remaining oils (extracted until 150 min), for which the acidity indices varied from about 2 to 2.6. The central composite nonfactorial design was used to optimise the extraction conditions, using the Statistica, version 5, software (Statsoft). The best results, in terms of oil recovered by SC-CO₂, were found at 19 MPa, 308 K and a superficial velocity of 6.0×10^–4 ms^–1.     

Variability of<Emphasis Type="Italic"> Satureja cuneifolia</Emphasis> Ten. essential oils and their antimicrobial activity depending on the stage of development

The essential oils obtained from Satureja cuneifolia Ten. harvested in the central part of Dalmatia at three ontogenetic stages were evaluated for their chemical composition and antimicrobial activity against food pathogens. The GC/MS analyses allowed 32 compounds to be determined; the main constituents of the essential oils were linalool (18.2–17.2%), carvacrol (16.0–5.0%), p-cymene (14.8–4.0%), -pinene (12.0–5.8%) and limonene (11.0–1.8%). The compounds linalool and borneol appeared to be relatively constant but carvacrol, limonene and -pinene showed variability during the growth cycles. The oils had a broad-spectrum antimicrobial activity against food pathogens in broth microdilution bioassays. Maximum activity was observed against the yeast Candida albicans, the Gram-negative bacteria Escherichia coli, Salmonella typhimurium and Proteus mirabilis and the Gram-positive bacteria Staphylococcus aureus and Bacillus cereus. The essential oils showed good antibacterial effects against E. coli with a minimum inhibitory concentration of 0.06% and a minimum bactericidal concentration of 0.12% during the flowering stage. These inhibitory effects are interesting in relation to the prevention of microbial contamination in many foods and, therefore, essential oils of S. cuneifolia could be used as substitutes for synthetic antimicrobial compounds.     

Chemical composition and antimicrobial activity of the essential oils of four Ocimum species growing in Tanzania

As part of ongoing research on Tanzanian plants used as edibles or spices, six samples of essential oils from four Ocimum species (O. basilicum, O. kilimandscharicum, O. lamiifolium, O. suave) were analyzed by GC and GC–MS. Eighty-one compounds, corresponding to 81.1–98.2% of the chemical components of the oils, were identified. Major compounds were either phenyl propane derivatives or terpenoids, including methyl eugenol, 1,8-cineole, camphor, bornyl acetate, germacrene-D, E-myroxide, germacrene-B, caryophylene oxide and p-cymene. The oils were also evaluated for antimicrobial activity against eight bacterial strains and three fungi. The oil of O. suave (B) showed the strongest antibacterial activity; O. suave (A), O. kilimandscharicum and, O. lamiifolium were moderately active, while O. basilicum oil was weakly active. However, none of the oils was active against the fungi species. The study has shown that, Ocimum oils could potentially be used as anti-infective agents.     

Environmental factors affect efficacy of some essential oils and resveratrol to control growth and ochratoxin A production by Penicillium verrucosum and Aspergillus westerdijkiae on wheat grain

This study determined the efficacy of three essential oils (bay, clove and cinnamon oil) and the antioxidant resveratrol (0–500 μg g⁻¹) on the control of growth and ochratoxin A (OTA) production by Penicillium verrucosum and Aspergillus westerdijkiae (=A. ochraceus) under different water activity (a_w, 0.90, 0.95, 0.995), and temperature (15, 25 °C) conditions on irradiated wheat grain. The most effective treatment (resveratrol) was then tested on natural grain. The ED₅₀ values for growth inhibition by the oils were 200–300 μg g⁻¹ at the a_w and the temperatures tested. For resveratrol, this varied from <50 μg g⁻¹ at 0.90–0.95 a_w to >350 at 0.995a_w at both temperatures. The ED₅₀ values for the control of OTA were slightly lower than for control of growth, with approx. 200 μg g⁻¹ required for the oils and 50–100 μg g⁻¹ of the antioxidant, at 0.90/0.95a_w and both temperatures. In wet grain (0.995a_w), higher concentrations were required. For growth there were statistically significant effects of single-, two- and three-way interactions between treatments except for concentration×temperature and concentration×temperature×essential oil/antioxidant treatment. For OTA control, statistically significant treatments were a_w, temperature×a_w, concentration×temperature, treatment×concentration, and three-way interaction of concentration×a_w×treatment for P. verrucosum and A. westerdijkiae. Subsequent studies were done with the best treatment (resveratrol, 200 μg g⁻¹) on natural wheat grain with either P. verrucosum or A. westerdijkiae at 0.85–0.995a_w and 15/25 °C over 28 days storage. This showed that the populations of the mycotoxigenic species and OTA contamination could be reduced by >60% by this treatment at the end of the storage period.     

Efficacy of heat treatment in the reduction of salmonellae and Escherichia coli O157:H– on alfalfa, mung bean and radish seeds used for sprout production

We studied the effect of hot-water treatment at various time/temperature regimes to design a decontamination process which is consistent with the recommendation of the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) to reduce pathogens on seeds by 5log cfu/g. Alfalfa, mung bean and radish seeds were inoculated by immersion with more than 10⁷ cfu/g of enterobacteria (Salmonella Senftenberg W775, S. Bovismorbificans and Escherichia coli O157:H^–), dried and stored at 2 °C. The numbers of salmonellae and E. coli O157:H^– on these seeds remained unchanged during storage for 8 weeks. To achieve sprouting rates of more than 95%, time-temperature regimes were defined. The thermal treatment of contaminated mung bean (2–20 min for 55–80 °C), radish and alfalfa seeds 0.5–8 min (53–64 °C) reduced all pathogens by more than 5log cfu/g. For S. Senftenberg W775 on radish seeds, D values of 3.2, 1.9 and 0.6 min were determined for exposure at 53, 55 and 58 °C and a z value of 6.2 °C was calculated. For alfalfa seeds, the respective D values were 3.0, 1.6, and 0.4 min and the z value was the same as that determined for radish seeds.     

Combination of warm water and hydrogen peroxide to reduce the numbers of Salmonella Typhimurium and Listeria innocua on field salad (Valerianella locusta)

The optimal conditions of decontamination of field salad by treatment with warm water containing hydrogen peroxide were determined with the aid of a central composite design. It was the aim to improve the hygienic status while keeping the sensory quality at the highest level possible. Two series of experiments were performed to study the effects of the following parameters: temperature (45–50 °C), H₂O₂ (1–4%) and treatment time (60–210 s). In series 1 the effect on sensory properties and reduction of total aerobic counts was investigated. To determine sensory quality, treated samples stored at 4 °C for 7 days were subjected to evaluation by 15 panelists. Overall visual quality (OVQ), color, texture and odor correlated well with product acceptability (R²=0.934, 0.946, 0.945 and 0.956, respectively), and OVQ was the most sensitive criterion. An acceptability of 75% in OVQ was chosen as the limit and an acceptance area was defined to deduce permitted conditions for decontamination. Within that area a reduction of total counts by 2.64–3.22 and 1.9–3.48 log₁₀ cfu/g was achieved after treatment and storage at 4 °C for 7 days, respectively. In series 2 field salad was inoculated with Listeria innocua and Salmonella Typhimurium and it was observed that within the area of acceptability a reduction of the numbers of the challenge organisms by 1.27–3.26 and 0.79–3.05 log₁₀ cfu/g, respectively, was achieved immediately after treatment, and by 1.64–3.09 (L. innocua) and 1.1–2.63 log₁₀ cfu/g (S. Typhimurium) after storage. The initial decontaminating effect on L. innocua and S. Typhimurium was sustained or even increased after storage, when the salad was treated below 48.9 °C for more than 68 s. Three independent experiments confirmed the reliability of the models of the count reduction of L. innocua and S. Typhimurium.     

Influence of organic acid concentration on survival of Listeria monocytogenes and Escherichia coli 0157:H7 in beef carcass wash water and on model equipment surfaces

Acid-adapted cultures of Escherichia coli O157:H7 and Listeria monocytogenes were inoculated in meat decontamination spray-washing runoff fluids in order to evaluate their survival and potential to form biofilms on stainless steel coupons. The cultures (10⁷ cfu ml⁻¹) and stainless steel coupons were exposed to mixtures of water and organic acid washings (composites of each of 2% acetic acid or lactic acid washings with water washings from meat decontamination in proportions of 1/9, 1/49, 1/99 [vol/vol]) or to water washings for up to 14 days at 15°C. E. coli O157:H7 formed biofilms and remained detectable (1.3 log cfu cm⁻²) on stainless steel for up to 4 d in the 1/9 dilution (pH 3.17–3.77) of the organic acid washings, and persisted throughout storage (14 d) in the 1/49 (pH 3.96–4.33) and 1/99 (pH 4.34–6.86) dilution of the organic acid washings. L. monocytogenes populations were unable to form detectable (<1.3 log cfu cm⁻²) biofilms in the 1/9 and 1/49 dilutions of both organic acid washings for up to 14 d; however, by day-14 in the 1/99 dilution of the washings, the pathogen was able to attach at detectable levels (2.7 to 3.4 logs). The pH effects of lower concentrations (1/49 or 1/99) of acidic washings decreased over time due to the formation of amine compounds produced by the natural meat flora, allowing resuscitation of the acid-stressed pathogen survivors. The resuscitation of acid-stressed pathogens may potentially enhance their survival and prevalence in biofilms and thus more attention should be focused on avoiding or minimizing the collection of decontamination runoff fluids on food contact equipment surfaces.     

Control of food spoiling bacteria in cooked meat products with nisin,lacticin 3147, and a lacticin 3147-producing starter culture

Nisin, in the form of the commercial product Nisaplin, and lacticin 3147 in whey powdered form were added to minced pork-meat in amounts of 0.15% (w/w) and 1.5% (w/w), respectively. The meat was cooked and inoculated with a Staphylococcus aureus strain of meat origin and a Listeria innocua strain at a level of 10⁷ or 10⁵ CFU g^–1. The batches were stored vacuum-packaged for 21 days at 8 °C. Nisin and lacticin 3147 immediately reduced the L. innocua population at the time of inoculation. Nisin showed higher inhibitory activity than lacticin 3147. During the storage period, a slight L. innocua growth was observed in the batches inoculated with the larger inoculum, and a bacteriostatic effect was observed against Listeria in the batches inoculated with 10⁵ CFU g^–1. Nisin maintained a constant S. aureus population in the cooked batch inoculated with 10⁷ CFU g^–1, although the bacteriocin was capable of reducing the amount of S. aureus by 90% in the batch inoculated with 10⁵ CFU g^–1. On the other hand, lacticin 3147 did not show an inhibitory effect against S. aureus in the cooked meat. The starter culture Lactococcus lactis DPC 303-T4 (containing the conjugative plasmid encoding production of lacticin 3147) was inoculated in a portion of a Longissimus dorsi pork muscle with brine. L. lactis DPC 303-T4 performed a good fermentation, but lacticin 3147 production was not found after 7 days at 12 °C of storage.     

Nisin treatments to control Listeria monocytogenes post-processing contamination on Anthotyros, a traditional Greek whey cheese, stored at 4°C in vacuum packages

Post-processing contamination and growth of Listeria monocytogenes in whey cheeses stored under refrigeration is an important safety concern. This study evaluated commercially available nisin (Nisaplin^®) as a biopreservative to control L. monocytogenes introduced post-processing on Anthotyros, a traditional Greek whey cheese, stored at 4°C in vacuum packages for up to 45 days. The whey used (pH 6.5–6.7) was from Feta cheese manufacture, and it was subjected either to natural acidification (pH 5.3, readjusted to 6.2 with 10% NaOH) prior to heating, or to direct acidification (pH 6.0–6.2) at 80°C with 10% citric acid. Nisin was added either to the whey (100 or 500 IU g⁻¹) prior to heating, or to the cheese (500 IU g⁻¹) prior to packaging, also inoculated with ca. 10⁴ cfu g⁻¹ of L. monocytogenes strain Scott A. In cheese samples without nisin, L. monocytogenes (PALCAM agar) exceeded 7 log cfu g⁻¹ after the first 10 days of storage, irrespective of the whey acidification method. All nisin treatments had an immediate lethal effect (0.7–2.2 log reduction) on L. monocytogenes populations at inoculation (day 0), which was more pronounced with 500 IU g⁻¹ added to the whey. This treatment also suppressed L. monocytogenes growth below the inoculation level for 30 and 45 days in naturally and directly acidified samples, respectively. All other treatments had weak antilisterial effects. Nisin reversed the natural spoilage flora of Anthotyros cheese from Gram-positive to Gram-negative, and this ecological alteration was far more pronounced in the most effective antilisterial treatments.     

Microbial and physicochemical succession in fermented sausages produced with bacteriocinogenic culture of Lactobacillus sakei and semi-purified bacteriocin mesenterocin Y

The influence of the bacteriocinogenic culture Lactobacillus sakei (10⁵/g) and semi-purified bacteriocin mesenterocin Y (2560 AU/kg) on the safety and quality of traditional Croatian fermented sausages was investigated. The addition of Lb. sakei and/or mesenterocin Y reduced microbial counts (P < 0.05) in the final products. After 28 days of ripening, coagulase-negative cocci decreased 1.5–2.0 log, yeasts 1.2–1.4 log and enterococci 1.7–2.7 log. In the case of the addition of Lb. sakei, the lactic acid bacteria count was significantly (P < 0.05) higher at day 7 of ripening, and was accompanied by a lower pH and a higher amount of lactic acid (P < 0.05). In the final product the amount of acetic acid was significantly lower. More intensive proteolysis and an increase in ammonia content were found at the beginning of fermentation, and in the second phase of ripening in the control samples, respectively. The free fatty acid concentration was significantly lower during the entire ripening process compared to the control (P < 0.05). Semi-purified mesenterocin Y did not affect the sensory properties of the sausages, whilst the addition of Lb. sakei enhanced them.     

Effects of γ-irradiation on molar mass and properties of Konjac mannan

Konjac mannan (KM) is a water-soluble glucomannan with high molar mass. Here, the effects of γ-irradiation on the structure of KM, its viscosity, molar mass distribution and the state of sorbed water were studied after irradiation at reduced pressure. These changes were investigated using ESR, FT-IR, UV, viscometer, SEC–MALS and DSC. Free radical yields increased with absorbed dose. Irradiation led to chain scission, but introduced no significant new chemical groups into the structure, apart from a small increase in content of carbonyl groups. The intrinsic viscosity, molar mass and radius of gyration decreased rapidly with increasing dose up to 10 kGy and then at a slower rate. The Mark–Houwink–Sakurada equation for the KM gave [η]=5.30×10⁻⁴ M^0.78. The α value showed that KM molecules are solvated in the form of random coils in water. There are three types of sorbed water in irradiated KM as in the original KM. There is no significant change in water binding ability for KM with M_W greater than 2×10⁵.     

Tunisian Salvia officinalis L. and Schinus molle L. essential oils: their chemical compositions and their preservative effects against Salmonella inoculated in minced beef meat

The essential oils (EOs) extracted from the aerial parts of cultivated Salvia officinalis L. and the berries of Schinus molle L. were analysed by gas chromatography–mass spectrometry (GC–MS) and 68 and 67 constituents were identified, respectively. The major constituents were 1,8-cineole (33.27%), β-thujone (18.40%), α-thujone (13.45%), borneol (7.39%) in S. officinalis oil and α-phellandrene (35.86%), β-phellandrene (29.3%), β-pinene (15.68%), p-cymene (5.43%) and α-pinene (5.22%) in S. molle oil.In its second part, the present study was conducted to evaluate the in vitro antimicrobial activity of both studied EOs. For this purpose, paper disc-diffusion method and broth microdilution test were used. The disc-diffusion method showed significant zone of lysis against all the pathogens studied (gram-negative and gram-positive bacteria, yeast). These activities remained stable after six months, and decreased approximately by 20% after one year of storage of the EOs at 4 to 7 °C. On comparing the efficiency of both EOs, S. officinalis EO exhibited higher antibacterial activity against the majority of strains and especially against Candida albicans (two fold more active according to the inhibition zones values). The minimal inhibitory concentrations (MICs) were reported between 4.5 mg/ml and 72 mg/ml on nutrient broth. The particular chemotype of each EO may be involved in its specific antimicrobial behaviour.Furthermore, the inhibitory effect of these EOs were evaluated against two foodborne pathogens belonging to Salmonella genus, experimentally inoculated (10³ CFU/g) in minced beef meat, which was mixed with different concentrations of the EO and stored at 4 to 7 °C for 15 days. Although the antibacterial activities of both EOs in minced beef meat were clearly evident, their addition had notable effects on the flavour and taste of the meat at concentrations more than 2% for S. molle and 1.5% for S. officinalis. One solution to the above-mentioned problem may be the use of combinations of different food preservation systems. In this context, each of the EOs has been used along with low water activity (addition of NaCl) in addition to low refrigeration temperatures. Results on the Salmonella growth showed that some combinations could be recommended to eliminate germs from minced raw beef. By using this method, a stable and, from a microbiological point of view, safe meat can be produced without substantial loss in sensory quality.Results obtained herein, may suggest that the EOs of S. officinalis and S. molle possess antimicrobial activity, and therefore, they can be used in biotechnological fields as natural preservative ingredients in food and/or pharmaceutical industry.     

Amyloglucosidase-catalyzed synthesis of n-octyl-d-glucoside—analysis using response surface methodology

Amyloglucosidase (3.2.1.3)-catalyzed synthesis of n-octyl-d-glucoside was optimized using response surface methodology. A central composite rotatable design involving 32 experiments of five variables at five levels was employed to study the glucosylation reaction. Among the variables studied, namely, n-octanol (15–75 MEq to d-glucose), enzyme (20–100 mg ), pH (4.0–8.0), buffer volume (0.2–1.0 ml) and temperature (30–70°C), amyloglucosidase concentration, pH and temperature were found to be significant. Experimental data fitted the second-order polynomial equation well, as indicated by an R² value of 0.895. Validation experiments carried out under predicted conditions showed good correspondence between experimental and predicted yields. Various surface plots were generated to describe the relationship between operating variables and the conversion yields. The highest yield of 53.5% predicted at optimum conditions of 75 Eq n-octanol, 20 mg amyloglucosidase, 0.2 ml, pH 7.8 buffer at 50 °C showed good correspondence to the experimental yield of 53.8% under these conditions.     

Effects of rapid chilling of carcasses and time of deboning on weight loss and technological quality of pork semimembranosus muscle

The effect of rapid air chilling of carcasses in the first 3 h of chilling at −31 °C (then at 2–4 °C, till 24 h post-mortem) and the possibility of earlier deboning (8 h post-mortem) after rapid air chilling, compared to conventional air chilling (at 2–4 °C, till 24 h post-mortem) on weight loss and technological quality (pH value, tenderness, drip loss, cooking loss and colour - L^*a^*b^* values) of pork M. semimembranosus was investigated. Under the rapid chilling conditions, weight loss was 0.8% at 8 h post-mortem and increased to 1.4% at 24 h post-mortem when weight loss was 2.0% under conventional chilling. Carcasses that were rapid chilled had significantly lower (P < 0.001) internal temperature in the deep leg at 4 (25.7 °C), 6 (13.0 °C), 8 (6.2 °C) and 24 h (3.8 °C) post-mortem compared to conventional chill treatment (32.7, 24.2, 19.1 and 5.1 °C, respectively). Rapid chilling reduced significantly (P < 0.05) the rate of pH value decline at 8 h (6.02) post-mortem in M. semimembranosus compared to conventional chill treatment (5.88). Compared to conventional chilling, in M. semimembranosus deboned in different time post-mortem, rapid chilling had a positive significant effect on drip loss (P < 0.05, muscles deboned 8 h post-mortem), cooking loss (P < 0.001) and incidence of pale colour (L^* value). Rapid chilling i.e. rapid chilling and earlier deboning had neither positive nor negative significant effects (P > 0.05) on other investigated technological quality parameters of M. semimembranosus (tenderness, a^* value and b^* value) compared to conventional chilling.     

Aggregation profile, preparation and nutritional characterization of African yam bean (Sphenostylis stenocarpa) acid and salt protein isolates

Aggregation tendencies of African yam bean (Sphenostylis stenocarpa) protein in the various aqueous extraction media with Ca²⁺, Mg²⁺ or at pI (isoelectric point) were determined. Water extractable S. stenocarpa protein aggregates more with addition of either Ca²⁺ or Mg²⁺ than at pI. In alkaline extractants considered (except at pH 10), aggregation tendency of the protein is in the order: pI > Ca²⁺ > Mg²⁺. The protein aggregation trend in salt media is a function of the salt type, aggregation in NaCl solution is of order: Ca²⁺ = pI > Mg²⁺, while in Na₂SO₄, we had pI > Mg²⁺ > Ca²⁺. S. stenocarpa protein aggregation was significantly (P < 0.05) more in Na₂SO₄ than NaCl. The amount of Ca²⁺ required for maximum precipitation of S. stenocarpa from alkaline (water-pH10) extractant was higher than that of Na₂SO₄ and more Ca-proteinate was obtained from the alkaline aliquot. The crude protein of the Ca-proteinates and isoelectric protein isolates obtained from salt and alkaline extract were in the range 71.7–91.8% (dry basis). Protein isolate from alkaline extract had significantly (P < 0.05) higher fat content than the one from salt extract. Isoelectrically precipitated isolates had lower ash content than Ca-proteinates. The percentage ratio of essential to non-essential amino acid was in the range 45–47%. With reference to FAO/WHO standard, the chemical score showed that most of the essential amino acid were in excess, thus, the amino acid distribution of S. stenocarpa protein isolates showed that it can fulfill the essential amino acid requirement of human especially the acid proteins and can be a good protein supplement in food and a wide range of new food products.     

Assessment of the microbiological safety of salad vegetables and sauces from kebab take-away restaurants in the United Kingdom

The purpose of this study was to establish the microbiological safety of salad vegetables and sauces served in kebab take-away restaurants. Comparison with published microbiological guidelines revealed that 4.7% of 1213 salad vegetable samples were of unsatisfactory microbiological quality due to Escherichia coli and/or Staphylococcus aureus levels at ≥10² cfu g⁻¹. Another 0.3% of salad samples were of unacceptable quality due to S. aureus at ≥10⁴ cfu g⁻¹ (2 samples) or the presence of Salmonella Kentucky (1 sample). Cucumber was the most contaminated salad vegetable with regards to unsatisfactory levels of E. coli (6.0%) or S. aureus (4.5%). Five percent of 1208 sauce samples were of unsatisfactory microbiological quality due to E. coli, S. aureus at ≥10² cfu g⁻¹ and/or Bacillus cereus and other Bacillus spp. at ≥10⁴ cfu g⁻¹. A further 0.6% of sauce samples were of unacceptable quality due to Bacillus spp. (Bacillus subtilis, Bacillus pumilus, Bacillus licheniformis) at ≥10⁵ cfu g⁻¹ or the presence of Salmonella Agbeni (1 sample). More samples of chilli sauce (8.7%) were of unsatisfactory or unacceptable microbiological quality than any other sauce types. The results emphasize the need for good hygiene practices in kebab take-away restaurants handling these types of ready-to-eat products.     

Reduction of soybean oligosaccharides and properties of alpha-D-galactosidase from Lactobacillus curvatus R08 and Leuconostoc mesenteroides [corrected] JK55

This study was undertaken to investigate the potential for reducing non-digestive oligosaccharides (NDO) in soy foods, as well as the influence of exogenous conditions on intracellular α-galactosidase (α-Gal) producing lactic acid bacteria. Two strains, Lactobacillus curvatus R08 and Leuconostoc mesenteriodes JK55, showed the highest levels of raffinose degrading activity at over 40 U mL⁻¹, and presented maximum activities during the stationary phase in a medium where raffinose was the only carbon source. Raffinose was the most effective inducer, followed by melibiose, and galactose; the enzymes were partially inhibited by fructose and sucrose. On the other hand, limited activity was observed in glucose. The strains displayed optimum activity levels at neutral pH and a 35–37 °C temperature range. The α-Gal activities of L. curvatus R08 and Leu. mesenteriodes JK55 were maintained at pH 6.5–10.0. The activity of the α-Gal enzyme was stable in a relatively broad range of temperatures from 0 to 40 °C for 3 h. In soymilk, Leu. mesenteriodes JK55 and L. curvatus R08 completely hydrolyzed the NDO after 18–24 h of fermentation. The abilities of L. curvatus R08 and Leu. mesenteriodes JK55 to degrade raffinose sugars and, particularly, to produce organic acids from sugar, could contribute to reductions in the anti-nutritional properties of soy, and to the accumulation of compounds with beneficial properties during food processing. Furthermore, this study provides the optimum conditions to induce α-Gal from these strains.     

Microbial, physical-chemical and sensory spoilage during the refrigerated storage of cooked pork loin processed by the sous vide method

The aim was to study spoilage during the refrigerated storage of cooked pork loin processed by the sous vide method. Samples were packaged under vacuum into polyamide-polypropylene pouches, cooked at an oven temperature/time of 70 °C/12 h, chilled at 3 °C and stored at 2 °C for 0, 5 or 10 weeks. Microbial (psychrotrophs, lactic acid bacteria, Enterobacteriaceae, moulds and yeasts), physical–chemical (pH, water activity, TBARS, acidity, L^*a^*b^* colour, texture profile analysis and shear force) and sensory (appearance, odour, flavour, texture and acceptance) parameters were determined. The results showed that sensory spoilage preceded microbiological spoilage of sous vide pork loin. Counts bellow 1 log cfu/g of psychrotrophs, anaerobic psychrotrophs, Enterobacteriaceae and lactic acid bacteria were detected in any control week, while moderate counts (2–3 log cfu/g) of moulds and yeasts were found. Minor changes in water activity, lipid oxidation, CIELab colour, hardness, cohesiveness or gumminess were associated with spoilage of pork loin, only decreases of lactic acid, springiness and shear force were observed. The pork loin was unacceptable after 10 weeks. This loss of acceptance was mainly due to the deterioration of meaty flavour and odour, although the loss of appearance, juiciness and firmness also contributed. Moderate warmed-over and rancidity were detected. The sensory analysis was the most effective method for determining the shelf life of the sous vide pork-based dishes.