Variation of glucosinolates in 62 varieties of Chinese cabbage (Brassica rapa L. ssp. pekinensis) and their antioxidant activity

Glucosinolate (GSL) and antioxidant activity in 62 varieties of Chinese cabbage (Brassica rapa L. ssp. pekinensis) were determined by HPLC and DPPH, HRSA, and FRAP assays. Five aliphatic GSLs: progoitrin, sinigrin, glucoalyssin, gluconapin, and glucobrassicanapin; four indolyl GSLs: 4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, and neoglucobrassicin; one aromatic GSL: gluconasturtiin were identified. Glucobrassicanapin and gluconapin documented the most abundant (average 4.52 and 3.72 μmol/g DW, respectively). The contents of total GSLs varied extensively among 62 varieties (range from 2.83 to 48.53 μmol/g DW). Comprehensive differences in total and individual GSL contents have also been observed among different varieties. Indolyl and aromatic GSL together accounted 26% of the total GSLs; but there are few differences among varieties. FC7 and FI17 could be good candidates for future breeding programs since they had a high quantity of glucobrassicin (2.10 and 1.66 μmol/g DW, respectively). Most of the Chinese cabbage varieties showed significant antioxidant activities when compare with positive control. However, three antioxidant assays were not significantly correlated with total GSLs. The presence of significant quantities of glucobrassicin in some varieties should be studied more extensively, since GSL is the precursor of indole-3-carbinol, a potent anticancer isothiocyanate.     

Evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of Vibrio parahaemolyticus in naturally contaminated seafood samples

We investigated the efficacy of a loop-mediated isothermal amplification (LAMP) assay for rapid screening of seafood samples naturally contaminated with Vibrio parahaemolyticus. A total of 171 seafood samples enriched in alkaline peptone water (APW) were assessed by LAMP assay and conventional culture methods, which consist of a combination of APW enrichment culture and plating onto CHROMagar Vibrio and TCBS agars. Compared with V. parahaemolyticus isolation using the conventional culture test, LAMP results showed 100% (30/30) and 90.8% (128/141) sensitivity and specificity, respectively. The conventional culture test required more than 3 days to isolate and identify V. parahaemolyticus in the APW enrichment culture. In contrast, the LAMP assay was markedly faster, requiring less than 60 min from the beginning of DNA extraction to final detection of V. parahaemolyticus. In total, the LAMP assay required 17-19 h from the beginning of enrichment culture to final determination. This is the first report of the LAMP assay for rapid screening of seafood samples naturally contaminated by V. parahaemolyticus.     

The use of a CYP51C gene based PCR-RFLP assay for simultaneous detection and identification of Fusarium avenaceum and F. tricinctum in wheat

Contamination of cereals with mycotoxins such as beauvericin (BEA), enniatins (Ens) and moniliformin (MON) is mainly caused by Fusarium avenaceum and F. tricinctum. This is a world-wide problem which requires rapid and sensitive detection methods. To allow for high throughput screening of large numbers of samples, a diagnostic PCR method was developed for the simultaneous detection of F. avenaceum and F. tricinctum. The interspecific divergence found in the Fusarium-specific CYP51C gene was used to design species-specific PCR primers. The specificity of the assay was demonstrated for DNA samples extracted from a wide range of Fusarium species belonging to the Fusarium head blight (FHB) complex, as well as for naturally-infected grain samples. The PCR-amplified products were digested with the restriction enzyme XbaI to enable differentiation between F. avenaceum and F. tricinctum. This PCR- restriction fragment length polymorphism (RFLP) assay proved to be a simple and relatively inexpensive method highly suited for routine detection and identification of F. avenaceum and F. tricinctum in wheat samples.     

Antibacterial activity of the extracts from the fruit rinds of Garcinia cowa and Garcinia pedunculata against food borne pathogens and spoilage bacteria

The crude hexane and chloroform extracts from the fruit rinds of Garcinia cowa and Garcinia pedunculata were studied for their antibacterial activity against some foodborne pathogens and spoilage bacteria such as Bacillus cereus, Bacillus coagulans, Bacillus subtilis, Staphylococcus aureus and Escherichia coli. The minimum inhibitory concentrations (MICs) of the extracts determined by the agar dilution method were ranging from 15 to 500 μg/ml and 300 to 1250 μg/ml for G. cowa and G. pedunculata, respectively. However, the hexane and chloroform extracts from the fruit rinds of G. cowa exhibited marked inhibitory effect against all the test organisms and were more effective than that of G. pedunculata extracts. The antibacterial activity of all the extracts was more pronounced against the tested Gram-positive bacteria than the tested Gram-negative bacterium. Furthermore, this study is the first report on the in vitro antibacterial activity of extracts from the fruit rinds of G. cowa and G. pedunculata.     

A bacteriocin-like substance produced from Lactobacillus pentosus 39 is a natural antagonist for the control of Aeromonas hydrophila and Listeria monocytogenes in fresh salmon fillets

We studied the ability of Lactobacillus pentosus 39, a BLS (Bacteriocin-like substance)-producing strain, to control the growth of Aeromonas hydrophila ATCC 14715 and Listeria monocytogenes ATCC 19117 artificially added to fresh salmon fillets at refrigeration temperatures and under simulated cold-chain break conditions.At refrigeration temperatures, Lb. pentosus 39 protective culture and its putative bacteriocin significantly reduced A. hydrophila counts compared with the control (2.1 and 1.4 log CFU/g reductions, respectively). Similar behaviour was observed for L. monocytogenes (3.6 and 1.3 log CFU/g reductions, respectively).Under simulated cold-chain break conditions, an increase in temperature (30°C for 12h) produced an evident increase in the development of A. hydrophila, L. monocytogenes, but also of Lb. pentosus 39, with a consequent increase in BLS production. This condition resulted in a greater reduction of both pathogens compared with samples stored at 4°C throughout the experiment (2.8 log CFU/g reduction for A. hydrophila, 5.8 log CFU/g reduction for L. monocytogenes). In samples treated with the putative bacteriocin alone, a less marked decrease was observed.Our study demonstrates the capability of Lb. pentosus 39 to control the growth of psychrotrophic bacteria in an experimental seafood model system. A similar biopreservation technology could provide more prolonged shelf-life during storage of ready-to-eat seafood, ensuring safety, even under extreme conditions.     

Integrated management of insect vectors of Aspergillus flavus in stored maize, using synthetic antioxidants and natural phytochemicals

The purpose of this study was to investigate the insecticidal activity of two benzoic acids 2(3)-tert-butyl-4 hydroxyanisole (BHA) and 2,6-di(tert-butyl)-p-cresol (BHT); two phenolic acids 3-phenyl-2-propenoic acid (CA) and trans-4-hydroxy-3-methoxycinnamic acid (FA) and two essential oils of Eugenia caryophyllata (clove tree) and Thymus vulgaris (thyme) against Sitophilus zeamais, Tribolium confusum and Rhyzopertha dominica, vector carriers of aflatoxigenic fungi in stored maize. The susceptibility of insects, the frequency of isolation of Aspergillus section Flavi in insects and maize, and the analysis of aflatoxin B₁ in maize were determined. BHA, BHT, BHA/BHT mixture and the natural phytochemicals AF and AF/AC mixture showed the highest insecticidal activity against S. zeamais, T. confusum and R. dominica after 120 days of incubation. The insecticidal efficacy of the volatile fraction of essential oils of clove and thyme showed less inhibition. There was no contamination of Aspergillus section Flavi in dead and live insects collected from maize treated with BHA. No aflatoxin B₁ accumulation was detected in the control and treatments. The information obtained shows that these substances have the potential to control pest insect vectors of aflatoxigenic fungi in stored maize in microcosms during 120 days.     

Characterisation of partially purified milk-clotting enzyme from Solanum dubium Fresen seeds

The seeds of Solanum dubium were blended and extracted using different types of buffers. The most reliable, quick, and efficient buffer was found to be 5% NaCl in acetate buffer (pH 5.0) which was used throughout the study. The extract was filtered and fractionated twice with ammonium sulphate. The partially purified enzyme was characterised by SDS–PAGE which showed a band of molecular weight of 66 kDa with the presence of other bands of low density. When compared with other plant enzymes, S. dubium enzyme was found to have higher clotting and proteolytic activities. The activity of the enzyme was steadily increased with enzyme and substrate concentration. The enzyme was found to be very stable against a wide range of pH values as well as a wide range of temperature (20–90 °C). The results of substrate specificity of the enzyme showed that the partially purified enzyme preferred both hydrophilic and hydrophobic amino acid residues at P1 position. The catalytic efficiency of the purified enzyme was enhanced by an aliphatic amino acids (Leu) compared to aromatic residue (Phe) at P1 position at the same site.     

Chemical analysis of edible aromatic plants growing in Tanzania

The volatiles from the aerial parts of edible plants growing in Tanzania, Leucas glabrata, Plectranthus laxiflorus, Salvia nilotica and Vernonia smithiana, were investigated by GC and GC/MS. Thirty-five compounds were identified from L. glabrata, representing 80.4% of the total oil; forty-three from P. laxiflorus (86.7%); twenty-four from S. nilotica (94.3%); and thirty-nine compounds from V. smithiana (92.9%). Among the identified components, menthone, (p + o)-cymene, trans-caryophyllene and caryophyllene oxide were found as the main ones. Furthermore, the essential oils were investigated for their antimicrobial activity as well as for their antiradical activity, through the DPPH method. Upon antimicrobial assays, the oil of V. smithiana showed very strong antimicrobial activity against Gram-positive bacteria, oral pathogens and pathogenic fungi; the oil of P. laxiflorus also exhibited strong activity, mostly against Gram-positive bacteria and especially oral pathogens, while L. glabrata showed strong activity against all assayed bacteria. The essential oil of S. nilotica appeared to have the most antioxidant activity but was almost inactive against all tested microorganisms.     

Varieties of antioxidant and antibacterial properties of Ecklonia stolonifera and Ecklonia kurome products harvested and processed in the Noto peninsula,Japan

Ecklonia stolonifera and Ecklonia kurome are traditional edible brown algae in the fisheries towns in Far East Asia. In the Noto Peninsula area, Ishikawa, Japan, both the Ecklonia are called ‘kajime’ and people believe that the algae improve the property of blood. To determine the varieties of antioxidant and antibacterial properties E. stolonifera and E. kurome products, assays for total phenolic content and antioxidant activities, including DPPH radical-scavenging activity, superoxide anion radical-generated by non-enzymatic system, ferrous-reducing power and WST-8, a tetrazolium salt, redox activity of four dried and two boiled E. stolonifera, and four dried and two raw E. kurome preparations were tested in this study. Furthermore, antibacterial activity of the products was tested. Though the total phenolic content, the antioxidant activity and the antibacterial activities of E. stolorifera and E. kurome products were high, these properties were varied by manufacturers or each product. Especially, two dried and two boiled products of E. stolonifera showed low activities. The results of this study indicate that the contents of functional compounds and their activities were affected or decreased greatly by the processing method.     

Phytochemical composition of the essential oils from three Apiaceae species and their antibacterial effects on food-borne pathogens

Essential oils (EOs) of three Apiaceae species, including Bunium persicum, Cuminum cyminum and Carum copticum, extracted by hydrodistillation, were analyzed by gas chromatography (GC) and GC/mass spectrometry. The main components of EOs of B. persicum were γ-terpinene (44.2%), cuminaldehyde (16.9%), γ-terpinen-7-al (10.5%), and ρ-cymene (8%). The major constituents of Cu. cyminum were cuminaldehyde (30.2%), ρ-cymene (14.1%), γ-terpinene (12.8%), and safranal (9.4%), while those of Ca. copticum were thymol (48.4%), ρ-cymene (21.8%) and γ-terpinene (21.3%). The antibacterial effects of the EOs were assessed on several food-borne pathogens, namely Staphylococcus aureus, Bacillus cereus, Escherichia coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes. The ranges of minimum inhibitory concentration (MIC) of the oils were 0.03–0.5, 0.18–3.0, and 0.37–3.0 mg/ml, respectively, for Ca. copticum, B. persicum and Cu. cyminum. Moreover, the combination of B. persicum and Cu. cyminum EOs confirmed synergistic and additive activities against the pathogens.     

Purification and characterization of intracellular proteinase from Lactobacillus casei ssp. casei LLG

The intracellular proteinase of Lactobacillus casei ssp. casei LLG was isolated in the cytoplasmic fraction with 0.05 M Tris-HCl buffer (pH 7.5). The enzyme was purified by the fast protein liquid chromatography system equipped with ion-exchange and gel filtration chromatographies. This proteinase comprised a single monomeric form and had a molecular weight of about 55 kDa and an isoelectric point near pH 4.9. The optimum pH and temperature for the enzyme activity were determined to be pH 6.5 and 37 degrees C, respectively. The enzyme was inactivated by metal-chelating compounds (EDTA, 1,10-phenanthroline) and less affected by serine proteinase inhibitors (diisopropylfluorophosphate, phenylmethylsulfonyl fluoride). Proteinase activity was increased by Ca++, Mn++, and Co++, and inhibited by Cu++, Mg++, and Zn++. The activity of this enzyme to hydrolyze casein appeared to be more active on beta-casein than alphas1-casein and kappa-casein as monitored by polyacrylamide gel electrophoresis.     

Comparative chemical composition,free radical-scavenging and cytotoxic properties of essential oils of six Stachys species from different regions of the Mediterranean Area

The chemical composition of essential oils of six Stachys species, S. cretica L. ssp. vacillans Rech. fil., S. germanica L., S. hydrophila Boiss., S. nivea Labill., S. palustris L. and S. spinosa L., obtained by hydrodistillation, was studied by GC and GC–MS. All the oils have in common a great percentage of fatty acids and esters (24.2–58.5%) and a high amount of sesquiterpenes (16–35.9%), with the exception of the oil from S. palustris, which consisted mainly of carbonylic compounds (25.4%). The antioxidant activity by DPPH test and the antiproliferative activity on a series of human cancer cell lines (C32, amelanotic melanoma and ACHN, renal cell adenocarcinoma) were investigated for all the oils. S. palustris,S. cretica and S. hydrophila showed the highest antiradical effect, with IC₅₀ values of 0.482, 0.652 and 0.664 mg/ml, respectively. The most antiproliferative essential oil against C32 cell line was the oil of S. germanica with a 77% of inhibition at a concentration of 100 μg/ml. S. germanica, S. palustris and S. spinosa showed the most antiproliferative activity on ACHN cell line, at a concentration of 100 μg/ml,with 81%, 77% and 73% inhibition, respectively.     

Evaluation of the antioxidant activity of three microalgal species for use as dietary supplements and in the preservation of foods

The antioxidant activity of the microalgal ethanolic extracts of Porphyridium cruentum, Phaeodactylum tricornutum and Chlorella vulgaris was determined by means of the β-carotene–linoleate model system. The results show that the activity of C. vulgaris extract was higher than those obtained for the other microalgal extracts tested and for the synthetics BHA (butylated hydroxyanisole), and BHT (butylated hydroxytoluene). In addition, the major constituents present in the ethanolic extracts of the three microalgae species were analyzed by means of GC and GC–mass spectrometry. The results showed that the tested microalgae may be an important source of natural antioxidants, as an alternative to higher plants or the production by chemical synthesis.     

In vitro antioxidant and antimicrobial activities of the extract of Pericarpium Citri Reticulatae of a new Citrus cultivar and its main flavonoids

To provide the base for the application of a new cultivar of Citrus reticulata Blanco, whose family is Rutaceae, a flavonoid extract of Pericarpium Citri Reticulatae (FEPCR), the dried rind of the ripe citrus, was obtained with 80% aq. ethanol. Total flavonoid content of FEPCR was determined by a colorimetric method. Total phenol content was estimated as gallic acid equivalents. The major constituents of FEPCR, including Hesperidin, Nobiletin and Tangeretin, were determined by HPLC analysis. The antioxidant activities of FEPCR, Hesperidin, Nobiletin and Tangeretin were evaluated by various antioxidant assays, including DPPH scavenging, hydroxyl radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging and reducing power. All samples showed antioxidant activities to some degree in all the tested methods. In addition to the antioxidant activity, the antimicrobial assay was measured as well. Six strains of microorganisms including Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Salmonella typhi and Enterobacter cloacae were used in the tests. FEPCR and Hesperidin displayed a broad antimicrobial spectrum and exerted antimicrobial effects in antimicrobial tests. But Tangeretin and Nobiletin exhibited low antimicrobial activities.     

Evaluation of antioxidant,antibacterial and anti-tyrosinase activities of four Macaranga species

The methanolic fresh leaf extracts of Macaranga gigantea, Macaranga pruinosa, Macaranga tanarius and Macaranga triloba were screened for their antioxidant properties (AOP), tyrosinase inhibition and antibacterial activities. Total phenolic content (TPC), 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging, ferric-ion reducing power (FRAP), ferrous-ion chelating (FIC) and lipid peroxidation inhibition (LPI) activities were used to evaluate the AOP. Modified 3,4-dihydroxy-L-phenylalanine (L-DOPA) method was used to determine tyrosinase inhibition activity, whereas antibacterial activity was determined using the disc-diffusion technique. TPC screening of the same species from different collection sites showed no significant difference between sites. M.triloba showed the highest ascorbic acid equivalent antioxidant activity (AEAC), FRAP and LPI values. M. tanarius, which showed the lowest TPC, AEAC, FRAP and LPI activities, exhibited the best FIC activity. M. pruinosa showed the best tyrosinase inhibition activity, whereas M. triloba showed the best antibacterial activity against Gram-positive bacteria species, with minimal inhibition dosage (MID) values as low as 10 μg/disc.     

Development and validation of a real-time PCR assay specific for Clostridium estertheticum and C. estertheticum-like psychrotolerant bacteria

A new real-time PCR assay was developed targeted to the psychrotolerant spoilage bacteria, Clostridum estertheticum, a causative agent of 'blown-pack' spoilage of vacuum packaged meats during chilled storage. Further, a robust validation of the sensitivity and specificity in different meat processing related matrices was carried out. Results show that real-time PCR is a valid method for the detection of C. estertheticum spores as long as consideration is given to the matrix being tested and the sensitivity of detection required. For meat, hide, blood/drip and environmental swabs it was possible to detect low numbers of C. estertheticum spores (approx 3 spores per ml) by direct real-time PCR (without pre-enrichment of the samples). For faeces and soil matrices, a cold temperature enrichment step was required prior to DNA extraction and real-time PCR analysis to increase the ability to detect samples containing C. estertheticum spores; this was particularly important when the samples contained low numbers of spores (less than 3 spores per ml). For matrices with high levels of PCR inhibitors such as soil, it was necessary to dilute the extracted DNA sample 100 fold especially for detection of high levels of contamination (greater than 10(3)per ml) otherwise a pre-enrichment was required.     

Composition and antimicrobial activity of the essential oils of three Satureja species growing in Tanzania

Hydro-distilled volatile oils from the aerial parts of three Satureja species were investigated, mainly by a combination of GC and GC/MS. One hundred and thirteen compounds were identified, representing 82.9–92.0% of the total oil. Among the identified components, spathulenol, cis-piperitone oxide, α-bisabolol oxide-B, terpinen-4-ol, linalool, bornyl acetate, β-bourbonene, isomenthone, thymol, neoisomenthol and menthone were found as the main components. Furthermore, the essential oils were investigated for their antimicrobial activity, by the agar dilution technique. The antimicrobial test results showed that the oils had a high antimicrobial activity against two Gram-positive and four Gram-negative bacteria, two oral pathogens and three pathogenic fungi. Gram-positive bacteria were more sensitive to the investigated oils than were Gram-negative bacteria. These results could support the suggestion of Satureja species as a source of antimicrobial ingredients for the food industry.     

Behaviour of the constitutive biota of two types of Spanish dry-sausages ripened in a pilot-scale chamber

The behaviour of the constitutive biota in eighty four samples belonging to two different types of Spanish dry-cured sausages during the ripening process in a pilot-scale chamber was investigated. Samples were analyzed in three stages during production: fresh product, first drying stage and finished product. Lactic acid bacteria (LAB) and Coagulase-negative cocci (CNC) were identified by the API system. In general, evolution of LAB and CNC during the ripening process of Spanish dry-cured sausages increased during the first days after which numbers of these organisms remained stable. Pediococcus pentosaceus and Staphylococcus xylosus, were the dominant species. Lactobacillus plantarum, Staphylococcus saprophyticus Staphylococcus simulans and Kocuria varians were also present. The results obtained show that the ripening process in a pilot-scale chamber under controlled conditions contributes to a more homogeneous behaviour of the constitutive biota, in comparison with commercial production standards.