In vitro anti-inflammatory properties of fermented pepino (Solanum muricatum) milk by γ-aminobutyric acid-producing Lactobacillus brevis and an in vivo animal model for evaluating its effects on hypertension.

BACKGROUND: The objectives of this study were to determine the in vitro anti-inflammatory and in vivo antihypertensive effects of fermented pepino (Solanum muricatum) milk by Lactobacillus brevis with the goal of developing functional healthy products. The inflammatory factors of fermented pepino milk with L. brevis were assessed in RAW 264.7 macrophages, including nitric oxide (NO) production. Inflammatory factor genes of cyclooxygenase (COX)-1 and -2, and tumor necrosis factor (TNF)-α were also assayed by a reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Results showed that fermented PE inhibited NO production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells with 150 mg mL(-1) fermented PE completely blocking LPS-induced NO production. The mRNA expressions of COX-1, COX-2, and iNOS were attenuated by treatment with higher concentrations of fermented PE (150 mg/ml). Cells treated with fermented pepino extract (PE) (100 ng mL(-1)) exhibited strikingly decreased LPS-induced expression of TNF-α mRNA. During the feeding trial, rats treated with 10% fermented pepino milk (100 µg 2.5 mL(-1)) and 100% fermented pepino milk (1000 µg 2.5 mL(-1)) exhibited significant decreases in the systolic blood pressure. CONCLUSION: Our results showed that fermented pepino milk has wide potential applications for development as a health food.

Improvement in antioxidant activity, angiotensin-converting enzyme inhibitory activity and in vitro cellular properties of fermented pepino milk by Lactobacillus strains containing the glutamate decarboxylase gene.

BACKGROUND: The purpose of this study was to evaluate the functional potential of fermented pepino extract (PE) milk by Lactobacillus strains containing the glutamate decarboxylase (GAD) gene. Three Lactobacillus strains were selected, including L. brevis BCRC 12310, L. casei BCRC 14082 and L. salivarius subsp. salivarius BCRC 14759. The contents of free amino acids, total phenolics content, total carotenoids and the associated functional and antioxidant abilities were analyzed, including angiotensin-converting enzyme (ACE) inhibition activity, 1,1-diphenyl-2-picylhydrazyl (DPPH) radical-scavenging ability and oxygen radical absorbance capacity (ORAC). Cell proliferation of fermented PE milk was also evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. RESULTS: Compared to the unfermented PE, fermented PE milk from Lactobacillus strains with the GAD gene showed higher levels of total phenolics, γ-aminobutyric acid, ACE inhibitory activity, DPPH, and ORAC. The viability of human promyelocytic leukemia cells (HL-60) determined by the MTT method decreased significantly when the cells were incubated with the PE and the fermented PE milk extracts. CONCLUSION: The consumption of fermented PE milk from Lactobacillus strains with the GAD gene is expected to benefit health. Further application as a health food is worthy of investigation. © 2012 Society of Chemical Industry.

The effects of Caulerpa microphysa enzyme-digested extracts on ACE-inhibitory activity and in vitro anti-tumour properties.

The aim of this study was to determine the ACE inhibitory activity and its anti-cancer properties of Caulerpa microphysa extracts. C. microphysa samples were digested with Flavourzyme, Alcalase, and pepsin. The ACE inhibitory activity of enzyme-digested C. microphysa decreased in the order of digestion with pepsin>Flavourzyme>Alcalase; that is, pepsin-extracted samples had significantly higher activity than the other enzyme extractions. To test its anti-tumour effects in vitro C. microphysa pepsin-digested extracts were applied to BALB/c mice with transplanted myelomonocytic leukaemia (WEHI-3) and Human promyelocytic leukaemia (HL-60) cell lines. The growth of both cell lines was inhibited, and extracts induced DNA damage, evaluated with a comet assay. The data demonstrate that C. microphysa pepsin-digested extract had the ability to anti-tumour effects. Further application as a health food is worthy of investigation.

Effect of antioxidant activity and functional properties of Chingshey purple sweet potato fermented milk by Lactobacillus acidophilus, L. delbrueckii subsp. lactis, and L. gasseri strains.

UNLABELLED: In this study, individual selected lactic acid bacteria strains Lactobacillus acidophilus (LA), L. delbrueckii subsp. lactis (LDL), and L. gasseri (LGA) were grown in Chingshey purple sweet potato (CPSP) substrate/media. CPSP is rich in anthocyanin, which possesses antioxidant activity and in vitro cell assay. The antioxidant ability and functional properties of the fermented milk were examined. High-performance liquid chromatographic (HPLC) method was used to analyze the free amino acid, organic acids, and anthocyanin content. Total phenolic compounds, scavenging effects of 1,1-diphenyl-2-picyl-hydrazyl (DPPH) radicals, and scavenging effects of superoxide anion radicals were determined to evaluate the antioxidant ability of the samples. The cell proliferation of the fermented PSP milk was evaluated by 3-(4,5-dimethyldiazol-2-yl)-2,5 diphenyl Tetrazolium Bromide (MTT) assay. The result indicated that the antioxidant ability of the fermented CPSP milk through LA, LDL, and LGA strains was significantly higher than CPSP. The main anthocyanins present in the samples are cyanidin and delphinidin. MTT assay has revealed that incubation with both PSP and fermented CPSP milk prevented the cell death of macrophage-like RAW264.7 cells. The potential health benefit of fermented PSP milk through LA, LDL, and LGA strains makes the further application of CPSP in health food highly worthwhile. PRACTICAL APPLICATION: (1) In our study, we have employed the γ-aminobutyric acid (GABA), organic acid contents, total phenol content, anthocyanins content, DPPH, oxygen radical absorbance capacity, superoxide dismutase activity assay, and cytotoxicity assay to assess the functional properties of fermented CPSP milk by different lactic acid bacteria. (2) Our results have revealed that the fermented CPSP milk samples possess high GABA concentrations, organic acid contents, anthocyanins contents, and antioxidant activity. This will provide potential opportunity to develop different functional food products from fermented CPSP milk. (3) The potential health benefit of fermented CPSP milk makes the further application of CPSP in health food highly worthwhile.

Phytochemicals characterization of solvent extracts from taro-scented japonica rice bran.

The major phytochemicals and antioxidant properties of taro-scented rice bran (TaiNung 71; TN71) extracts using 3 different solvents are characterized. Some progress is realized in creating an economic value for rice bran that has long been considered an agricultural waste. Various solvent extracts reveal the presence of phenolic compounds, oryzanols, tocopherols, and tocotrienols. Ethyl acetate (EtOAc) can extract more oryzanols (1.55 ± 0.20 g/kg rice bran). Meanwhile, the methanol (MeOH) extract possesses a higher yield in total contents (15.42 ± 1.41 g/kg bran), which includes phenolic compounds (2.69 ± 0.29 g gallic acid equivalent/kg bran), tocopherols (251 ± 26 mg/kg bran) and tocotrienols (111 ± 4 mg/kg bran). The MeOH extract exhibits more effective antioxidant activity against various oxidative systems in vitro, including the inhibition of linoleic acid peroxidation (33.89%), scavenging of DPPH radicals (83.88%), and reducing power. It is found that the yield, total content in phenolic compounds and tocols of the extracts increase with increasing Synder's polarity value and viscosity, which can then be used as the indices in isolation of the desired rice bran phytochemicals extracts.

Characteristics of virulent vibrio parahaemolyticus isolated from Oregon and Washington.

Thirty-four virulent strains of Vibrio parahaemolyticus containing tdh and/or trh genes isolated from Oregon and Washington coastal water were analyzed for O-group antigens and urease activity, and by pulsed-field gel electrophoresis. Six O serotypes (O1, O3, O4, O5, O10, and O11) were identified among the isolates, with the O5 group (19 isolates) being the most prevalent, followed by the 01 group (9 isolates). Nearly all (33 of 34) isolates were capable of producing urease, which reaffirmed the correlation between urease production and virulence factors of V. parahaemolyticus strains isolated from the Pacific Northwest. Pulsed-field gel electrophoresis analysis with NotI and SfiI digestions of the 34 V. parahaemolyticus isolates plus five clinical strains revealed 22 patterns (NlS1 to N20S22), with NIS1 (25.6%) being the most common, followed by N2S2 (10.3%). Nine Oregon isolates were grouped with a 1997 Oregon outbreak strain (027-1C1) with the same serotype (O5), virulence factors (tdh+ and trh+), and genotype (N S 1). Three Washington isolates were found to share the same serotype (O1), virulence factors (tdh' and trh'), and genotype (N2S2) with a 1997 Washington outbreak strain (10293). The repetitive isolation of virulent strains of V. parahaemolyticus identical to clinical strains involved in previous outbreaks indicates potential hazards associated with oyster consumption. These data may be useful in risk assessment of V. parahaemolyticus infections associated with raw oyster consumption in Oregon and Washington.

A pulsed field gel electrophoresis (PFGE) study that suggests a major world-wide clone of Salmonella enterica serovar Enteritidis.

Since human infections by Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) have been increasing world-wide over the past years and epidemiological studies have implicated the consumption of meat, poultry, eggs and egg products, elucidation of the predominant subtypes for this Salmonella spp. is important. In this study, 107 poultry and food isolates of Salmonella Enteritidis obtained from Germany were analyzed by pulsed field gel electrophoresis (PFGE), and the subtypes were compared with those of the 124 human isolates obtained in Taiwan. Results showed that for these 107 poultry and food isolates, when XbaI, SpeI and NotI were used for chromosomal DNA digestion followed by PFGE analysis, a total of 19, 20 and 19 PFGE patterns, respectively, were identified. Of them, 51 (47.7%), 52 (48.6%) and 42 (39.3%) strains belong to a single pattern of X3, S3 and N3, respectively, and 34 strains belong to a pattern combination of X3S3N3, which was the major subtype. When PFGE patterns of these 107 German isolates were compared with those of the 124 human isolates obtained in Taiwan, pattern combination of X3S3N3 was found as the most common pattern shared by isolates from both areas. PT4 is a major phage type for German and Taiwan isolates. Although most of the X3S3N3 strains are of this phage type, some strains of other PFGE patterns are also of this phage type. Since strains used in this study were unrelated, i.e., they were isolated from different origins in areas geographically far apart from each other, the PFGE study suggests a major world-wide clone of S. enterica serovar Enteritidis.

Characterisation of antimicrobial resistance patterns and class 1 integrons among Escherichia coli and Salmonella enterica serovar Choleraesuis strains isolated from humans and swine in Taiwan.

Escherichia coli isolates from humans (n=110) and swine (n=61) and Salmonella enterica serovar Choleraesuis isolates (n=95) from swine in southern Taiwan were characterised for antimicrobial resistance patterns and class 1 integrons. All E. coli isolates and S. Choleraesuis isolates were multidrug resistant and demonstrated high resistance to beta-lactams, aminoglycosides, tetracycline, sulfonamides, spectinomycin, chloramphenicol and nalidixic acid. By polymerase chain reaction and DNA sequencing, 104 (61%) E. coli isolates and 31 (33%) S. Choleraesuis isolates were found to carry class 1 integrons. The gene cassette array dfrA12-orfF-aadA2 was the most prevalent (24%) among the human and swine E. coli isolates, whilst the gene cassette array dfrA12-orfF-aadA2-sul1 was the most prevalent (24%) among S. Choleraesuis strains. For E. coil isolates, all class 1 integrons were located on conjugated plasmids. Meanwhile, human and swine E. coli isolates carrying identical gene cassettes were genetically unrelated. Our results revealed that multidrug resistance and class 1 integrons were widely present in E. coli and S. Choleraesuis isolates obtained in Taiwan and that class 1 integrons might play an important role in contributing to the horizontal transfer of antimicrobial resistance.

Development of PCR primers for the detection of Salmonella enterica serovar Choleraesuis based on the fliC gene.

Salmonella enterica serovar Choleraesuis may cause swine salmonellosis and human infection. Because the conventional method for detection of this Salmonella serovar may take 3 to 5 days, a PCR method for detection was evaluated. By comparing the sequence of the phase 1 flagellin (fliC) gene of Salmonella Choleraesuis with that of other Salmonella serovars and of other bacteria species available in GenBank, two PCR primers (flinC-F and flinC-R) were designed. Using these primers, all 97 Salmonella Choleraesuis strains assayed generated the expected PCR product, with a molecular mass of 963 bp. Except for S. enterica Paratyphi C, Salmonella isolates other than Salmonella Choleraesuis and non-Salmonella isolates, including strains of Enterobacteriaceae, all generated negative PCR results. Salmonella Paratyphi C could be differentiated from Salmonella Choleraesuis through the use of primers designed from the viaB gene. When Salmonella Choleraesuis isolates from swine stool, pork, liver, feed, and human whole blood samples were assayed with a preenrichment step, as low as 1 CFU/g or ml of the original sample could be detected.