• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1472-1478
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• 2004

The effects of microorganisms inoculated in beef, pork and chicken on the production of various biogenic amines (BA) were examined. Acinetobacter haemolyticus, Aeromonas hydrophila subsp. hydrophila, Alcaligenes faecalis subsp. faecalis, Bacillus cereus, Bacillus subtilis, Enterobacter aerogenes, Enterobacter cloacae, Escherichia coli, Lactobacillus alimentarius, Lactobacillus curvatus, Leuconostoc mesenteroides subsp. Mesenteroides, Proteus mirabilis, Proteus vulgaris, Pseudomonas aerugina, Salmonella enteritidis and Salmonella typhimurium were inoculated into beef, pork and chicken and incubated for 24 h at optimum temperatures of each bacterium. In ground beef, total amount of amines (TAA) produced was highest in the sample inoculated with Bacillus cereus, followed by Enterobacter cloacae. In ground pork, TAA was highest in the sample inoculated with Alcaligenes faecalis, followed by Enterobacter cloacae, Proteus vulgaris and Bacillus cereus. TAA of chicken breast was highest in the sample inoculated with Alcaligenes faecalis, followed by Bacillus cereus and Lactobacillus alimentarius while in chicken leg was the sample inoculated with Proteus vulgaris, followed by Enterobacter aerogenes, Enterobacter cloacae and Alcaligenes faecalis. Among biogenic amines produced, cadaverine (CAD) was detected at the highest level, followed by putrescine (PUT) and tyramine (TYM), their order being reversed by the kind of microorganism in beef and pork. In chicken breast and leg, CAD level was still the highest but PUT, TYM or PHM was the second highest, depending upon the kind of microorganism inoculated. In total, Alcaligenes faecalis, Enterobacter cloacae and Bacillus cereus were ones that produced a larger amount of BAs regardless of meat sources from different species.

• Food Science of Animal Resources
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• v.38 no.1
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• pp.152-161
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• 2018

This study aims to evaluate biogenic amine levels of bez sucuks (BS) produced with different meat:fat ratios. For this, three BS groups were manufactured with meat:fat ratios of 90:10 (BS10), 80:20 (BS20), and 70:30 (BS30). The pH and water activity values and biogenic amine amounts of sucuk samples were determined during processing and storage periods and the pH values of the initial mixtures of BS samples were in the range 5.51-5.74, decreasing to 4.72-4.94 by the $14^{th}$ day. The water activity values of BS samples showed significant decreases as a result of the drying stage and reached to range 0.913-0.935 on the $14^{th}$ day of processing (p<0.05). Although BS10 had the highest tyramine (434.12 mg/kg), histamine (5.69 mg/kg), cadaverine (12.48 mg/kg), putrescine (17.83 mg/kg), 2-phenylethylamine (15.43 mg/kg), and tryptamine (122.41 mg/kg) levels at the end of processing stage (p<0.05), spermine and spermidine levels did not differ between the BS samples due to their utilization of different meat:fat ratios (p>0.05). Similarly, the tryptamine (205.11 mg/kg), putrescine (43.57 mg/kg), and tyramine (766.23 mg/kg) levels of BS10 were higher than BS20 and BS30 samples at the end of storage (p<0.05). The results showed that BS10 with the highest meat ratio had the highest tryptamine, putrescine, and tyramine levels at the end of the processing and storage period.

• Toxicological Research
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• v.31 no.3
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• pp.299-305
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• 2015

An HPLC analytical method was validated for the quantitative determination of biogenic amines in agricultural products. Four agricultural foods, including apple juice, Juk, corn oil and peanut butter, were selected as food matrices based on their water and fat contents (i.e., non-fatty liquid, non-fatty solid, fatty liquid and fatty solid, respectively). The precision, accuracy, recovery, limit of detection (LOD) and quantification (LOQ) were determined to test the validity of an HPLC procedure for the determination of biogenic amines, including tryptamine, ${\beta}$-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine, in each matrix. The LODs and LOQs for the biogenic amines were within the range of 0.01~0.10 mg/kg and 0.02~0.31 mg/kg, respectively. The relative standard deviation (RSD) of intraday for biogenic amine concentrations ranged from 1.86 to 5.95%, whereas the RSD of interday ranged from 2.08 to 5.96%. Of the matrices spiked with biogenic amines, corn oil with tyramine and Juk with putrescine exhibited the least accuracy of 84.85% and recovery rate of 89.63%, respectively, at the lowest concentration (10 mg/kg). Therefore, the validation results fulfilled AOAC criteria and recommendations. Subsequently, the method was applied to the analysis of biogenic amines in fermented agricultural products for a total dietary survey in Korea. Although the results revealed that Korean traditional soy sauce and Doenjang contained relatively high levels of histamine, the amounts are of no concern if these fermented agricultural products serve as condiments.

• Parasites, Hosts and Diseases
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• v.51 no.3
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• pp.279-287
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• 2013

Autophagy is a process of cytoplasmic degradation of endogenous proteins and organelles. Although its primary role is protective, it can also contribute to cell death. Recently, autophagy was found to play a role in the activation of host defense against intracellular pathogens. The aims of our study was to investigate whether host cell autophagy influences Toxoplasma gondii proliferation and whether autophagy inhibitors modulate cell survival. HeLa cells were infected with T. gondii with and without rapamycin treatment to induce autophagy. Lactate dehydrogenase assays showed that cell death was extensive at 36-48 hr after infection in cells treated with T. gondii with or without rapamycin. The autophagic markers, LC3 II and Beclin 1, were strongly expressed at 18-24 hr after exposure as shown by Western blotting and RT-PCR. However, the subsequent T. gondii proliferation suppressed autophagy at 36 hr post-infection. Pre-treatment with the autophagy inhibitor, 3-methyladenine (3-MA), down-regulated LC3 II and Beclin 1. The latter was also down-regulated by calpeptin, a calpain inhibitor. Monodansyl cadaverine (MDC) staining detected numerous autophagic vacuoles (AVs) at 18 hr post-infection. Ultrastructural observations showed T. gondii proliferation in parasitophorous vacuoles (PVs) coinciding with a decline in the numbers of AVs by 18 hr. FACS analysis failed to confirm the presence of cell apoptosis after exposure to T. gondii and rapamycin. We concluded that T. gondii proliferation may inhibit host cell autophagy and has an impact on cell survival.

• Korean Journal of Microbiology
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• v.55 no.3
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• pp.258-267
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• 2019

The purpose of this study was to investigate the effect of lactic acid bacteria (LAB) on the growth and biogenic amines (BA) formation of Enterobacter aerogenes CIH05 in skim milk and soymilk. Lactobacillus acidophilus GK20, Lactobacillus paracasei GK74, and Lactobacillus plantarum GK81 isolated from mustard kimchi did not produce BA in the decarboxylation broth. L. paracasei GK74 exhibited the highest cell viability and antimicrobial compounds producing ability in fermented skim milk and soymilk samples, while the lowest producer was L. plantarum GK81. The production yield of lactic acid, hydrogen peroxide, and bacteriocin was dependent on the species of Lactobacillus and the type of culture medium. As LAB the number of viable cells of E. aerogenes CIH05 were higher in skim milk than in soymilk. When mixed culture with L. acidophilus GK20 and L. paracasei GK74 and treated with bacteriocin solution (300 AU/ml) obtained from these strains in milk media, the cell growth and cadaverine and histamine contents of E. aerogenes CIH05 were significantly (P < 0.05) lower than the respective values in control sample.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.51 no.6
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• pp.667-672
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• 2018

We investigated the quality of 12 commercial salted and fermented anchovy Engraulis japonicas sauces by measuring their chemical compositions, bacteria concentrations, and biogenic amine contents. The sauces had a moisture content of 67.13-69.83% (mean: 68.17%), salinity of 20.00-25.84% (mean: 22.29%), pH of 5.14-6.28 (mean: 5.63), volatile basic nitrogen content of 119.12-273.37 mg/100 g (mean: 199.19 mg/100 g), total nitrogen content of 0.82-1.50% (mean: 1.30%) and amino nitrogen content of 550.17-1,086.62 mg/100 g (mean: 774.99 mg/100 g). The viable cell counts ranged from not detected to $1.6{\times}10^3CFU/mL$, and the number of biogenic amine-forming bacteria was very low or undetectable. The tested samples contained 372.32-2,111,61 mg/kg (mean 813.48 mg/kg) histamine, 29.62-144.29 mg/kg (mean 98.14 mg/kg) cadaverine, 87.89-530.84 mg/kg (mean 329.91 mg/kg) tryptamine, 20.89-127.17 mg/kg (mean 60.49 mg/kg) putrescine, and 13.08-109.91 mg/kg (mean 57.74 mg/kg) tyramine. Whereas no spermidine or spermine was detected in any sample. These results strongly suggest the necessary of monitoring the biogenic amine contents of commercial salted and fermented anchovy sauces carefully to ensure consumer health.

• Journal of Microbiology and Biotechnology
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• v.31 no.7
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• pp.949-955
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• 2021

Previously, our research group isolated Bifidobacterium breve IDCC4401 from infant feces as a potential probiotic. For this study, we evaluated the safety of B. breve IDCC4401 using genomic and phenotypic analyses. Whole genome sequencing was performed to identify genomic characteristics and investigate the potential presence of genes encoding virulence, antibiotic resistance, and mobile genetic elements. Phenotypic analyses including antibiotic susceptibility, enzyme activity, production of biogenic amines (BAs), and proportion of D-/L-lactate were evaluated using E-test, API ZYM test, high-performance liquid chromatography (HPLC), and D-/L-lactic acid assay respectively. The genome of B. breve IDCC4401 consists of 2,426,499 bp with a GC content of 58.70% and 2,016 coding regions. Confirmation of the genome as B. breve was provided by its 98.93% similarity with B. breve DSM20213. Furthermore, B. breve IDCC4401 genes encoding virulence and antibiotic resistance were not identified. Although B. breve IDCC4401 showed antibiotic resistance against vancomycin, we confirmed that this was an intrinsic feature since the antibiotic resistance gene was not present. B. breve IDCC4401 showed leucine arylamidase, cystine arylamidase, α-galactosidase, β-galactosidase, and α-glucosidase activities, whereas it did not show production of harmful enzymes such as β-glucosidase and β-glucuronidase. In addition, B. breve IDCC4401 did not produce any tyramine, histamine, putrescine, cadaverine, or 2-phenethylamine, which are frequently detected BAs during fermentation. B. breve IDCC4401 produced 95.08% of L-lactate and 4.92% of D-lactate. Therefore, our findings demonstrate the safety of B. breve IDCC 4401 as a potential probiotic for use in the food industry.

• Journal of Applied Biological Chemistry
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• v.63 no.4
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• pp.375-385
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• 2020

The purpose of this study was to investigate the quality characteristics of kimchi prepared with a single starter culture of biogenic amines (BA)-forming lactic acid bacteria (LAB) or a combined starter cultures composed of BA-forming and BA-degrading LAB. As the fermentation proceeded, the lactic acid bacterial count, titratable acidity, and BA content in kimchi prepared with myeolchi-aekjeot were slightly higher than those of kimchi prepared with saeu-jeot. The amount and type of BA produced by LAB were mostly strain dependent rather than species specific. Among all of the isolated LAB strains, the highest levels of cadaverine, histamine, putrescine and tyramine were produced by Leuconostoc mesenteroides MBK32, Lactobacillus brevis MBK34, Lactobacillus curvatus MBK31 and Enterococcus faecalis SBK31, respectively. BA-forming and BA-degrading starter cultures played an important role in the growth rate and organic acid-producing ability of LAB in kimchi. Interestingly, BA contents in kimchi increased by adding single BA-forming LAB starter were effectively lowered by the mixed cultures with BA-degrading LAB.

• Journal of the Korean Applied Science and Technology
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• v.37 no.6
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• pp.1535-1544
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• 2020

The primary objective of this study was to isolate and identify amine oxidase-producing probiotic Bacillus strains from traditional fermented soybean paste. Biogenic amines (BA)-forming bacteria isolated from the samples were identified as Bacillus sp. TS09, Bacillus licheniformis TS17, Bacillus subtilis TS19, Bacillus cereus TS23, Bacillus sp. TS30, Bacillus megaterium TS31, B. subtilis TS44, Bacillus coagulans TS46 and Bacillus amyloliquefaciens TS59. Meanwhile, B. subtilis TS04 and TS50 isolated from the same samples exhibited good probiotic properties, including the tolerance to artificial gastric juice and bile salts, the adhesion to intestinal epithelial cells, and the production of bacteriocin(s) active against BA-forming bacteria (Bacillus sp. TS30 and B. subtilis TS44). In addition, the amine oxidase produced by B. subtilis TS04 and TS50 significantly decreased the formation of BA, especially cadaverine, putrescine, and tyramine, therefore, these strains could be considered good potential probiotic candidates to prevent or reduce BA accumulation in food products.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.25-32
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• 2021

Inflammatory reactions activated by lipopolysaccharide (LPS) of gram-negative bacteria can lead to severe septic shock. With the recent emergence of multidrug-resistant gram-negative bacteria and a lack of efficient ways to treat resulting infections, there is a need to develop novel anti-endotoxin agents. Antimicrobial peptides have been noticed as potential therapeutic molecules for bacterial infection and as candidates for new antibiotic drugs. We previously designed the 9-meric antimicrobial peptide Pro9-3 and it showed high antimicrobial activity against gram-negative bacteria. Here, to further examine its potency as an anti-endotoxin agent, we examined the anti-endotoxin activities of Pro9-3 and elucidated its mechanism of action. We performed a dye-leakage experiment and BODIPY-TR cadaverine and limulus amebocyte lysate assays for Pro9-3 as well as its lysine-substituted analogue and their enantiomers. The results confirmed that Pro9-3 targets the bacterial membrane and the arginine residues play key roles in its antimicrobial activity. Pro9-3 showed excellent LPS-neutralizing activity and LPS-binding properties, which were superior to those of other peptides. Saturation transfer difference-nuclear magnetic resonance experiments to explore the interaction between LPS and Pro9-3 revealed that Trp3 and Tlr7 in Pro9-3 are critical for attracting Pro9-3 to the LPS in the gram-negative bacterial membrane. Moreover, the anti-septic effect of Pro9-3 in vivo was investigated using an LPS-induced endotoxemia mouse model, demonstrating its dual activities: antibacterial activity against gram-negative bacteria and immunosuppressive effect preventing LPS-induced endotoxemia. Collectively, these results confirmed the therapeutic potential of Pro9-3 against infection of gram-negative bacteria.