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1.
Appl Environ Microbiol ; 87(10)2021 04 27.
Article in English | MEDLINE | ID: mdl-33674429

ABSTRACT

Bacterial vaginosis (BV) is the most common vaginal disorder of reproductive-aged women, yet its etiology remains enigmatic. One clinical symptom of BV, malodor, is linked to the microbial production of biogenic amines (BA). Using targeted liquid chromatography mass spectrometry, we analyzed 149 longitudinally collected vaginal samples to determine the in vivo concentrations of the most common BAs and then assessed their relationship to BV and effect upon the growth kinetics of axenically cultured vaginal Lactobacillus species. Increases in cadaverine, putrescine, and tyramine were associated with greater odds of women transitioning from L. crispatus-dominated vaginal microbiota to microbiota that have a paucity of Lactobacillus spp. and from Nugent scores of 0 to 3 to Nugent scores of 7 to 10, consistent with BV. Exposure to putrescine lengthened the lag time and/or slowed the growth of all vaginal Lactobacillus spp. except L. jensenii 62G. L. iners AB107's lag time was lengthened by cadaverine but reduced in the presence of spermidine and spermine. The growth rate of L. crispatus VPI 3199 was slowed by cadaverine and tyramine, and strain-specific responses to spermine and spermidine were observed. BAs were associated with reduced production of d- and l-lactic acid by vaginal Lactobacillus spp., and this effect was independent of their effect upon Lactobacillus species growth. The exceptions were higher levels of d- and l-lactic acid by two strains of L. crispatus when grown in the presence of spermine. Results of this study provide evidence of a direct impact of common biogenic amines on vaginal Lactobacillus spp.IMPORTANCELactobacillus spp. are credited with providing the primary defense against gynecological conditions, including BV, most notably through the acidification of the vaginal microenvironment, which results from their production of lactic acid. The microbial production of BAs has been hypothesized to play a mechanistic role in diminishing Lactobacillus species-mediated protection, enabling the colonization and outgrowth of diverse anaerobic bacterial species associated with BV. Here, we demonstrate that in vivo increases in the most commonly observed BAs are associated with a loss of Lactobacillus spp. and the development of BV, measured by Nugent score. Further, we show that BAs formed by amino acid decarboxylase enzymes negatively affect the growth of type strains of the most common vaginal Lactobacillus spp. and separately alter their production of lactic acid. These results suggest that BAs destabilize vaginal Lactobacillus spp. and play an important and direct role in diminishing their protection of the vaginal microenvironment.


Subject(s)
Biogenic Amines/biosynthesis , Lactobacillus/metabolism , Vaginosis, Bacterial/microbiology , Female , Humans , Lactic Acid/biosynthesis , Lactobacillus/growth & development , Vagina/microbiology
2.
Molecules ; 25(22)2020 Nov 22.
Article in English | MEDLINE | ID: mdl-33266479

ABSTRACT

Cheese ripening involves a number of biochemical processes, mainly of a proteolytic nature, which are initially triggered principally by milk-coagulating enzymes and, afterward, by microorganisms or enzymes of microbial origin. The proteolytic reactions affect, primarily, the synthesis of macro- and medium-molecular peptides from casein. In turn, the advanced proteolysis ends in the formation of short peptides and free amino acids. Further reactions may lead to the formation of nutritionally unfavorable biogenic amines. The present study aimed to determine changes in the contents of bioactive peptides (anserine and L-carnosine), free amino acids, and biogenic amines throughout the ripening of cheese models produced with the addition of Lactobacillus genus bacteria. The contents of amino acids varied considerably in the cheese models, depending on the bacterial strain added and ripening time. After five weeks of ripening, the total content of free amino acids in the cheese models ranged from 611.02 (a cheese model with Lactobacillus casei 2639) to 1596.64 mg kg-1 (a cheese model with Lb. acidophilus 2499). After the same time, the contents of the total biogenic amines in the cheese models with the addition of lactobacilli were lower than in the control cheese model (except for the model with Lb. rhamnosus 489). Anserine was detected in all cheese models (79.29-119.02 mg kg-1), whereas no L-carnosine was found over a five-week ripening period in the cheese models with Lb. delbrueckii 490 and Lb. casei 2639. After a five-week ripening, the highest total content of bioactive peptides was determined in the cheese models containing Lb. acidophilus 2499 (136.11 mg kg-1).


Subject(s)
Amino Acids/analysis , Biogenic Amines/analysis , Cheese/microbiology , Lactobacillus/metabolism , Peptides/analysis , Amino Acids/biosynthesis , Amino Acids/chemistry , Animals , Biogenic Amines/biosynthesis , Biogenic Amines/chemistry , Carnosine/analysis , Carnosine/metabolism , Cheese/analysis , Fermentation , Food Handling/methods , Food Microbiology/methods , Food Quality , Milk/chemistry , Milk/microbiology , Netherlands , Peptides/chemistry , Peptides/metabolism , Proteolysis , Time Factors
3.
Curr Microbiol ; 76(10): 1095-1104, 2019 Oct.
Article in English | MEDLINE | ID: mdl-31270565

ABSTRACT

Fermented cereals are part of the main traditional diets of many people in Africa, usually obtained from artisanal production. The intensification of their manufacturing, responding to the consumers demand, requires a better control to ensure their sanitary, nutritional, and taste qualities, hence, the need of selecting accurate and safe starter cultures. In the present study, 48 lactic acid bacteria (LAB) strains, previously isolated from Algerian fermented wheat lemzeiet, were analyzed for different technological properties. 14 LAB strains, belonging to Pediococcus pentosaceus, Enterococcus faecium, Lactobacillus curvatus, Lactobacillus brevis, and Leuconostoc mesenteroides species, decreased rapidly the pH of the flour extract broth close to 4 or below. 91% of strains showed extracellular protease activity, but only 12% were amylolytics. 18 LAB strains inhibited or postponed the growth of three fungal targets Rhodotorula mucilaginosa UBOCC-A-216004, Penicillium verrucosum UBOCC-A-109221, and Aspergillus flavus UBOCC-A-106028. The strains belonging to Lactobacillus spp., Leuconostoc fallax, L. mesenteroides, and Weissella paramesenteroides were the most antifungal ones. Multiplex PCR for biogenic amines' production did not reveal any of the genes involved in the production of putrescine, histamine, and tyramine for 17 of the 48 strains. The obtained results provided several candidates for use as starter culture in the future production of lemzeiet.


Subject(s)
Fermented Foods/microbiology , Food Microbiology , Lactobacillales/isolation & purification , Lactobacillales/metabolism , Triticum/microbiology , Amylases/metabolism , Antifungal Agents/pharmacology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Biogenic Amines/biosynthesis , DNA, Bacterial/genetics , Fermentation , Hydrogen-Ion Concentration , Lactobacillales/enzymology , Lactobacillales/genetics , Peptide Hydrolases/metabolism
4.
Metab Eng ; 47: 184-189, 2018 05.
Article in English | MEDLINE | ID: mdl-29477859

ABSTRACT

Biocatalytic C-H amination is one of the most challenging tasks. C-H amination reaction can hardly be driven efficiently by solely one enzyme so far. Thus, enzymatic synergy represents an alternative strategy. Herein, we report an "Artificially Bioamination Pathway" for C-H amination of cyclohexane as a model substrate. Three enzymes, a monooxygenase P450BM3 mutant, an alcohol dehydrogenase ScCR from Streptomyces coelicolor and an amine dehydrogenase EsLeuDH from Exiguobacterium sibiricum, constituted a clean cascade reaction system with easy product isolation. Two independent cofactor regeneration systems were optimized to avoid interference from the endogenous NADH oxidases in the host E. coli cells. Based on a stepwise pH adjustment and ammonium supplement strategy, and using an in vitro mixture of cell-free extracts of the three enzymes, cyclohexylamine was produced in a titer of 14.9 mM, with a product content of up to 92.5%. Furthermore, designer cells coexpressing the three required enzymes were constructed and their capability of alkane bio-amination was examined. This artificially designed bioamination paves an attractive approach for enzymatic synthesis of amines from accessible and cheap alkanes.


Subject(s)
Alkanes/metabolism , Biogenic Amines/biosynthesis , Escherichia coli , Amination , Bacillales/enzymology , Bacillales/genetics , Bacterial Proteins/biosynthesis , Bacterial Proteins/genetics , Cytochrome P-450 Enzyme System/biosynthesis , Cytochrome P-450 Enzyme System/genetics , Escherichia coli/genetics , Escherichia coli/metabolism , Streptomyces coelicolor/enzymology , Streptomyces coelicolor/genetics
5.
J Appl Microbiol ; 122(4): 1078-1091, 2017 Apr.
Article in English | MEDLINE | ID: mdl-28117533

ABSTRACT

AIMS: The tyraminogenic potential of the strains Enterococcus faecalis EF37 and ATCC 29212 was investigated in a synthetic medium containing defined amounts of tyrosine and phenylalanine at different temperatures. METHODS AND RESULTS: Enterococci growth and the production of biogenic amines (BA) were evaluated in relation to their pre-growth in medium containing tyrosine. Significant differences between the two strains were evidenced at metabolic level. Both the pre-adapted strains grew faster in all the tested conditions, independently of the presence of the precursor. Temperatures of 30 and 40°C positively affected the growth parameters. The tyrosine decarboxylase (tyrDC) activity of the strain EF37 was positively affected by pre-adaptation, while ATCC 29212 showed a faster and higher tyramine accumulation with not-adapted cells. The expression analysis of the gene tyrDC confirmed the influence of the growth conditions on gene transcription. CONCLUSIONS: The small differences found between the two strains in the maximum transcript level reached rapidly after the inoculum and the different behaviour in the tyramine accumulation suggested the possible involvement of complex regulation mechanisms on the tyrDC or on the membrane transport systems, which could affect the different BA accumulation trend. SIGNIFICANCE AND IMPACT OF THE STUDY: This study gives deeper insight into the metabolic regulation of tyrDC activity of enterococci.


Subject(s)
Biogenic Amines/biosynthesis , Enterococcus faecalis/metabolism , Tyrosine Decarboxylase/biosynthesis , Culture Media , Enterococcus faecalis/enzymology , Enterococcus faecalis/genetics , Enterococcus faecalis/growth & development , Phenylalanine/metabolism , Transcription, Genetic , Tyramine/biosynthesis , Tyrosine/metabolism , Tyrosine Decarboxylase/genetics
6.
Curr Microbiol ; 74(11): 1337-1342, 2017 Nov.
Article in English | MEDLINE | ID: mdl-28761980

ABSTRACT

The effect of disodium fumarate (DF) on the ruminal fermentation profiles, the accumulation of lipopolysaccharide (LPS) and bioamines, and the composition of the ruminal bacterial community was investigated by in vitro rumen fermentation. The addition of DF increased the total gas production; the concentrations of propionate, valerate, total volatile fatty acids, and ammonia-nitrogen; and the rumen pH after a 24 h fermentation. By contrast, DF addition decreased the ratio of acetate to propionate and the concentrations of lactate, lipopolysaccharide, methylamine, tryptamine, putrescine, histamine, and tyramine (P < 0.05). Principal coordinates analysis and molecular variance analysis showed that DF altered the ruminal bacterial community (P < 0.05). At the phylum level, DF decreased the proportion of Proteobacteria, and increased the proportions of Spirochaetae and Elusimicrobia (P < 0.05). At the genus level, DF decreased the percentage of Ruminobacter, while increasing the percentage of Succinivibrio and Treponema (P < 0.05). Overall, the results indicate that DF modified rumen fermentation and mitigated the production of several toxic compounds. Thus, DF has great potential for preventing subacute rumen acidosis in dairy cows and for improving the health of ruminants.


Subject(s)
Bacteria/metabolism , Biogenic Amines/biosynthesis , Fermentation/drug effects , Fumarates/pharmacology , Lipopolysaccharides/biosynthesis , Microbiota/drug effects , Rumen/microbiology , Animals , Bacteria/classification , Bacteria/genetics , Cattle , Cluster Analysis , Metagenome , Metagenomics/methods
7.
Food Microbiol ; 62: 92-98, 2017 Apr.
Article in English | MEDLINE | ID: mdl-27889172

ABSTRACT

We assessed the safety of 49 Tetragenococcus halophilus strains isolated from doenjang in Korea. Minimum inhibitory concentration assays showed that all strains can be considered as susceptible to ampicillin, erythromycin, penicillin G, tetracycline, and vancomycin, but resistant to ciprofloxacin based on the Enterococcus breakpoint values provided by the European Committee on Antimicrobial Susceptibility testing in 2015. Ciprofloxacin resistance was sufficiently high to consider the potential for acquisition of transmissible determinants. Two strains exhibiting potentially acquired resistance to chloramphenicol and gentamicin, and chloramphenicol alone, were identified. None of the strains exhibited α-hemolytic activity or biofilm formation; two strains exhibited weak ß-hemolytic activity. Doenjang isolates produced an average of 3338.6 ppm of tyramine in the laboratory, considerably higher than the levels produced by two reference strains. All of the test strains exhibited similar cadaverine, histamine, and putrescine production patterns. Most T. halophilus strains could grow at a NaCl concentration >18%, exhibited acid production at 15% NaCl, and expressed strain-specific protease and lipase activities. The potential acquisition of transmissible determinants for antibiotic resistance and tyramine production identified in this study necessitate the need for a thorough safety assessment of T. halophilus before it can be considered for use in food fermentation processes.


Subject(s)
Enterococcaceae/isolation & purification , Fermentation , Food Safety , Glycine max/microbiology , Soy Foods/microbiology , Anti-Bacterial Agents/pharmacology , Biofilms/growth & development , Biogenic Amines/biosynthesis , Bioreactors , Ciprofloxacin/pharmacology , Drug Resistance, Microbial , Enterococcaceae/drug effects , Enterococcaceae/genetics , Food Handling , Food Microbiology , Histidine Decarboxylase/genetics , Microbial Sensitivity Tests , Republic of Korea , Sodium Chloride, Dietary , Tyrosine Decarboxylase/genetics
8.
Molecules ; 20(6): 9510-23, 2015 May 26.
Article in English | MEDLINE | ID: mdl-26016543

ABSTRACT

Most red wines commercialized in the market use the malolactic fermentation process in order to ensure stability from a microbiological point of view. In this second fermentation, malic acid is converted into L-lactic acid under controlled setups. However this process is not free from possible collateral effects that on some occasions produce off-flavors, wine quality loss and human health problems. In warm viticulture regions such as the south of Spain, the risk of suffering a deviation during the malolactic fermentation process increases due to the high must pH. This contributes to produce wines with high volatile acidity and biogenic amine values. This manuscript develops a new red wine making methodology that consists of combining the use of two non-Saccharomyces yeast strains as an alternative to the traditional malolactic fermentation. In this method, malic acid is totally consumed by Schizosaccharomyces pombe, thus achieving the microbiological stabilization objective, while Lachancea thermotolerans produces lactic acid in order not to reduce and even increase the acidity of wines produced from low acidity musts. This technique reduces the risks inherent to the malolactic fermentation process when performed in warm regions.The result is more fruity wines that contain less acetic acid and biogenic amines than the traditional controls that have undergone the classical malolactic fermentation.


Subject(s)
Kluyveromyces/metabolism , Lactic Acid/metabolism , Malates/metabolism , Saccharomycetales/metabolism , Schizosaccharomyces/metabolism , Wine/analysis , Acetic Acid/analysis , Acetic Acid/metabolism , Biogenic Amines/analysis , Biogenic Amines/biosynthesis , Fermentation , Humans , Hydrogen-Ion Concentration , Spain , Vitis/metabolism , Vitis/microbiology
9.
J Sci Food Agric ; 95(6): 1157-62, 2015 Apr.
Article in English | MEDLINE | ID: mdl-24976408

ABSTRACT

BACKGROUND: Biogenic amines have gained a great deal of attention due to their toxic potential in humans. Carp is one of the most important freshwater fish species in China. Salt and sugar are capable of preserving food. There is a limited amount of information on the changes of biogenic amines in freshwater fish influenced by salt and sugar. This study aimed to detect the changes in biogenic amines, sensory attributes, total volatile basic nitrogen (TVB-N) and total viable counts (TVC) of carp influenced by different concentrations of salt and sugar stored at 4 °C. RESULTS: TVB-N and TVC increased with storage time, which was in accordance with the changes of sensory scores. The eight biogenic amines were detected in fresh carp. Putrescine and cadaverine were the main biogenic amines found in carp fillets stored at 4 °C; they had a significant (P < 0.05) correlation with TVB-N. CONCLUSION: Salt processing was found to inhibit the increase of TVB-N, TVC, putrescine and cadaverine in carp. High salt concentration had a positive effect on extending the shelf-life of the carp, compared to low salt concentration.


Subject(s)
Biogenic Amines/biosynthesis , Carbohydrates , Carps , Cold Temperature , Food Preservation , Seafood/analysis , Sodium Chloride , Animals , Cadaverine/biosynthesis , Diet , Food Microbiology , Food Preservatives , Food Storage/methods , Humans , Nitrogen/metabolism , Putrescine/biosynthesis , Seafood/microbiology , Seafood/standards , Taste
10.
J Sci Food Agric ; 95(11): 2215-21, 2015 Aug 30.
Article in English | MEDLINE | ID: mdl-25263002

ABSTRACT

BACKGROUND: Microbial transglutaminase (MTGase) is an enzyme utilized in the food industry in many areas. In this study, the suppression effect of MTGase at various levels (0, 2, 5, 10 g kg(-1)) on the formation of biogenic amines in mackerel was determined during refrigerated storage of 8 days. RESULTS: Mince added with 2 g kg(-1) MTGase showed the lowest formation of putrescine, cadaverine and tyramine throughout the storage. Histamine exceeded the consumable limit (500 mg kg(-1) ) after the 4th day, except for that containing 2 g kg(-1) MTGase. The formation of total volatile basic nitrogen and total free amino acid content were retarded and the pH value was unaltered by addition of MTGase. With increasing MTGase levels, the growth of total psychrophilic bacteria, mould, yeast and coliform bacteria was retarded. The sensory scores of mackerel mince increased as MTGase concentrations increased. CONCLUSION: MTGase plays a role in maintaining the quality of mackerel mince during refrigerated storage. As a result of the present study, a new use for MTGase in the food industry is revealed. It will contribute especially in the field of development of products for consumers with allergic sensitivity.


Subject(s)
Biogenic Amines/biosynthesis , Food Handling/methods , Food Microbiology , Food Preservation/methods , Perciformes , Seafood/analysis , Transglutaminases/metabolism , Amino Acids/metabolism , Animals , Bacteria/drug effects , Cold Temperature , Colony Count, Microbial , Fungi/drug effects , Humans , Nitrogen/metabolism , Refrigeration , Seafood/microbiology , Seafood/standards , Taste , Transglutaminases/pharmacology
11.
J Ind Microbiol Biotechnol ; 41(10): 1553-62, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25085740

ABSTRACT

As one of the major microbes in the soy sauce fermentation, Candida versatilis enriches the flavor and improves the quality of soy sauce. In this study, a combination of five different GC-MS and LC-MS-based metabolome analytical approaches was used to analyze the intracellular, extracellular and whole metabolites of C. versatilis. Our results found out that a total of 132, 244 and 267 different metabolites were detectable from the intracellular, extracellular and whole part, respectively. When exposed to 0. 9 and 18 % salt, respectively, 114, 123 and 129 different intracellular metabolites, 184, 200 and 178 extracellular metabolites and 177, 188 and 186 whole metabolites were detected, respectively. Our data showed that salt enhances the metabolic capacity of C. versatilis, especially its amino acid and enhances the synthesis and secretion of some metabolites of C. versatilis, especially the aldehydes and phenols, such as vanillin, guaiacol and 5-hydroxymethylfurfural. Our data also showed that special attention has to be paid to the generation of biogenic amines when C. versatilis was treated with salt.


Subject(s)
Candida/metabolism , Metabolome , Sodium Chloride/metabolism , Stress, Physiological , Benzaldehydes/metabolism , Biogenic Amines/biosynthesis , Furaldehyde/analogs & derivatives , Furaldehyde/metabolism , Guaiacol/metabolism , Principal Component Analysis , Salt Tolerance
12.
Molecules ; 19(11): 18139-51, 2014 Nov 06.
Article in English | MEDLINE | ID: mdl-25383752

ABSTRACT

Rice produces a wide array of phytoalexins in response to pathogen attacks and UV-irradiation. Except for the flavonoid sakuranetin, most phytoalexins identified in rice are diterpenoid compounds. Analysis of phenolic-enriched fractions from UV-treated rice leaves showed that several phenolic compounds in addition to sakuranetin accumulated remarkably in rice leaves. We isolated two compounds from UV-treated rice leaves using silica gel column chromatography and preparative HPLC. The isolated phenolic compounds were identified as phenylamide compounds: N-trans-cinnamoyltryptamine and N-p-coumaroylserotonin. Expression analysis of biosynthetic genes demonstrated that genes for arylamine biosynthesis were upregulated by UV irradiation. This result suggested that phenylamide biosynthetic pathways are activated in rice leaves by UV treatment. To unravel the role of UV-induced phenylamides as phytoalexins, we examined their antimicrobial activity against rice fungal and bacterial pathogens. N-trans-Cinnamoyltryptamine inhibited the growth of rice brown spot fungus (Bipolaris oryzae). In addition to the known antifungal activity to the blast fungus, sakuranetin had antimicrobial activity toward B. oryzae and Rhizoctonia solani (rice sheath blight fungus). UV-induced phenylamides and sakuranetin also had antimicrobial activity against rice bacterial pathogens for grain rot (Burkholderia glumae), blight (Xanthomonas oryzae pv. oryzae) and leaf streak (X. oryzae pv. oryzicola) diseases. These findings suggested that the UV-induced phenylamides in rice are phytoalexins against a diverse array of pathogens.


Subject(s)
Anti-Infective Agents , Ascomycota/growth & development , Biogenic Amines , Burkholderia/growth & development , Plant Leaves , Rhizoctonia/growth & development , Ultraviolet Rays , Xanthomonas/growth & development , Anti-Infective Agents/chemistry , Anti-Infective Agents/isolation & purification , Anti-Infective Agents/metabolism , Anti-Infective Agents/pharmacology , Biogenic Amines/biosynthesis , Biogenic Amines/chemistry , Biogenic Amines/isolation & purification , Biogenic Amines/pharmacology , Gene Expression Regulation , Oryza/chemistry , Oryza/metabolism , Plant Leaves/chemistry , Plant Leaves/metabolism
13.
J Sci Food Agric ; 94(8): 1537-42, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24154976

ABSTRACT

BACKGROUND: This study aimed to enhance and improve the quality and safety of soy sauce. In the present work, the change of biogenic amines, such as histamine, tyramine, cadaverine, spermidine, was examined by the treatment of Candida versatilis and Zygosaccharomyces rouxii, and the influence of salt-tolerant yeast on biogenic amines was analysed during the whole fermentation process. RESULTS: The results showed that the content of biogenic amines was elevated after yeast treatment and the content of biogenic amines was influenced by using yeast. The dominating biogenic amine in soy sauce was tyramine. At the end of fermentation, the concentrations of biogenic amines produced by Zygosaccharomyces rouxii and Candida versatilis in the soy mash were 122.71 mg kg(-1) and 69.96 mg kg(-1) . CONCLUSIONS: The changes of biogenic amines in high-salt liquid soy mash during fermentation process indicated that a variety of biogenic amines were increased in the fermentation ageing period, which may be due to amino acid decarboxylation to form biogenic amines by yeast decarboxylase. The fermentation period of soy sauce should be longer than 5 months because biogenic amines began to decline after this time period.


Subject(s)
Biogenic Amines/biosynthesis , Candida/metabolism , Fermentation , Food Handling/methods , Soy Foods , Zygosaccharomyces/metabolism , Biogenic Amines/analysis , Cadaverine/analysis , Candida/growth & development , Histamine/analysis , Hydrogen-Ion Concentration , Soy Foods/analysis , Soy Foods/microbiology , Spermidine/analysis , Time Factors , Tyramine/analysis , Zygosaccharomyces/growth & development
14.
World J Microbiol Biotechnol ; 30(8): 2213-21, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24668182

ABSTRACT

Using decarboxylation medium and 16S rDNA sequence analysis, histamine-producing bacteria (HPB) in blue scad (Decapterus maruadsi) were isolated and identified, and the histamine-producing abilities of the isolated HPB were determined. Nine mesophilic strains (H1-H9) isolated from the muscle of blue scad were identified as the genera of HPB, including Arthrobacter bergeri (H1), Pseudomonas sp. (H2, H5 and H6), Psychrobacter sp. (H3), Shewanella baltica (H4 and H7), and Aeromonas salmonicida (H8 and H9), respectively. Results showed that most of the HPB strains were weak on histamine formation (13.0-20.4 mg/l), except for the H8 strain with the ability of producing 115 mg of histamine/l in trypticase soy broth containing 1.0 % L-histidine. As the strongest HPB in blue scad, bacterial strain H8 also presented a strong ability to produce other biogenic amines, such as putrescine, cadaverine, spermidine, spermine, tyramine and tryptamine. Therefore, the H8 strain identified as the genus of A. salmonicida was the dominant mesophilic HPB strain for producing histamine and other biogenic amines in blue scad at room temperature.


Subject(s)
Gram-Negative Bacteria/classification , Gram-Negative Bacteria/isolation & purification , Histamine/biosynthesis , Muscles/microbiology , Perciformes/microbiology , Animals , Bacterial Typing Techniques , Biogenic Amines/biosynthesis , Perciformes/anatomy & histology , Perciformes/classification , Phylogeny
15.
Food Microbiol ; 35(1): 10-4, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23628608

ABSTRACT

The development of fast, reliable and culture-independent molecular tools to detect bacteria producing biogenic amines deserves the attention of research and ultimately of the food industry in order to protect consumers' health. Here we present the application of a simple, low-cost, fast and sensitive method to perform microdroplet-based multiplex PCR, directly on a food matrix, for the simultaneous detection of bacterial genes involved in biogenic amine biosynthesis. After inoculating wine with Lactobacillus brevis IOEB 9809, cell lysis and DNA amplification are performed in one single step, without preliminary nucleic acid extraction or purification treatments. The assay is performed in about 30 min, requiring 150 nL of starting sample and it enables the detection of down to 15 bacterial cells. With respect to traditional culture techniques, the speed, the simplicity and the cheapness of this procedure allow an effective monitoring of microbial cells during food-making and processing.


Subject(s)
Biogenic Amines/biosynthesis , Food Contamination/analysis , Levilactobacillus brevis/isolation & purification , Multiplex Polymerase Chain Reaction/methods , Food Microbiology , Genes, Bacterial , Levilactobacillus brevis/genetics , Microfluidic Analytical Techniques/methods , Wine/microbiology
16.
Food Microbiol ; 36(2): 335-42, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24010615

ABSTRACT

This work is the first large-scale study on vineyard-associated yeast strains from Apulia (Southern Italy). Yeasts were identified by Internal Transcribed Spacer (ITS) ribotyping and bioinformatic analysis. The polymorphism of interdelta elements was used to differentiate Saccharomyces cerevisiae strains. Twenty different species belonging to 9 genera were identified. Predominant on the grape surface were Metschnikowia pulcherrima, Hanseniaspora uvarum and Aureobasidium pullulans, whereas M. pulcherrima and H. uvarum were dominant in the early fermentation stage. A total of 692 S. cerevisiae isolates were identified and a number of S. cerevisiae strains, ranging from 26 to 55, was detected in each of the eight fermentations. The strains were tested for biogenic amines (BAs) production, either in synthetic media or grape must. Two Pichia manshurica, an Issatchenkia terricola and a M. pulcherrima strains were able to produce histamine and cadaverine, during must fermentation. The production of BAs in wine must was different than that observed in the synthetic medium. This feature indicate the importance of an "in grape must" assessment of BAs producing yeast. Overall, our results suggest the importance of microbiological control during wine-making to reduce the potential health risk for consumer represented by these spoilage yeasts.


Subject(s)
Biodiversity , Biogenic Amines/biosynthesis , Vitis/microbiology , Wine/microbiology , Yeasts/isolation & purification , Yeasts/metabolism , Biogenic Amines/toxicity , Culture Media/metabolism , Fermentation , Food Safety , Italy , Vitis/metabolism , Yeasts/classification , Yeasts/genetics
17.
J Dairy Res ; 80(1): 122-7, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23298547

ABSTRACT

The aim of the study was to identify the species of Enterobacteriaceae present in Montasio cheese and to assess their potential to produce biogenic amines. Plate count methods and an Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) approach, combined with 16S rDNA sequencing, were used to investigate the Enterobacteriaceae community present during the cheesemaking and ripening of 6 batches of Montasio cheese. Additionally, the potential decarboxylation abilities of selected bacterial isolates were qualitatively and quantitatively assessed against tyrosine, histidine, ornithine and lysine. The most predominant species detected during cheese manufacturing and ripening were Enterobacter cloacae, Escherichia coli and Hafnia alvei. The non-limiting physico-chemical conditions (pH, NaCl% and a(w)) during ripening were probably the cause of the presence of detectable levels of Enterobacteriaceae up to 120 d of ripening. The HPLC test showed that cadaverine and putrescine were the amines produced in higher amounts by almost all isolates, indicating that the presence of these amines in cheese can be linked to the presence of high counts of Enterobacteriaceae. 44 isolates produced low amounts of histamine (<300 ppm), and four isolates produced more than 1000 ppm of this amine. Only 9 isolates, belonging to the species Citrobacter freundii, Esch. coli and Raoultella ornithinolytica, appeared to produce tyramine. These data provided new information regarding the decarboxylase activity of some Enterobacteriaceae species, including Pantoea agglomerans, Esch. fergusonii and R. ornithinolytica.


Subject(s)
Aromatic-L-Amino-Acid Decarboxylases/metabolism , Cheese/microbiology , Enterobacteriaceae/classification , Enterobacteriaceae/enzymology , Animals , Bacterial Load , Biogenic Amines/biosynthesis , Cadaverine/biosynthesis , Cattle , Chromatography, High Pressure Liquid , Fermentation , Histamine/biosynthesis , Italy , Putrescine/biosynthesis , Tyramine/biosynthesis
18.
BMC Microbiol ; 12: 247, 2012 Oct 31.
Article in English | MEDLINE | ID: mdl-23113922

ABSTRACT

BACKGROUND: Ingestion of fermented foods containing high levels of biogenic amines (BA) can be deleterious to human health. Less obvious is the threat posed by BA producing organisms contained within the food which, in principle, could form BA after ingestion even if the food product itself does not initially contain high BA levels. In this work we have investigated the production of tyramine and putrescine by Lactobacillus brevis IOEB 9809, of wine origin, under simulated gastrointestinal tract (GIT) conditions. RESULTS: An in vitro model that simulates the normal physiological conditions in the human digestive tract, as well as Caco-2 epithelial human cell lines, was used to challenge L. brevis IOEB 9809, which produced both tyramine and putrescine under all conditions tested. In the presence of BA precursors and under mild gastric stress, a correlation between enhancement of bacterial survival and a synchronous transcriptional activation of the tyramine and putrescine biosynthetic pathways was detected. High levels of both BA were observed after exposure of the bacterium to Caco-2 cells. CONCLUSIONS: L. brevis IOEB 9809 can produce tyramine and putrescine under simulated human digestive tract conditions. The results indicate that BA production may be a mechanism that increases bacterial survival under gastric stress.


Subject(s)
Biogenic Amines/biosynthesis , Levilactobacillus brevis/metabolism , Stress, Physiological , Caco-2 Cells , Epithelial Cells/microbiology , Gene Expression Profiling , Humans , Levilactobacillus brevis/isolation & purification , Microbial Viability , Models, Biological , Putrescine/biosynthesis , Tyramine/biosynthesis , Wine/microbiology
19.
J Appl Microbiol ; 113(6): 1407-16, 2012 Dec.
Article in English | MEDLINE | ID: mdl-22963007

ABSTRACT

AIMS: The capacity of Lactobacillus sakei CRL1862 to prevent the growth of pathogens and its ability to degrade sarcoplasmic and myofibrillar proteins in pork meat systems was evaluated. In addition, basic safety aspects of Lact. sakei CRL1862 such as production of biogenic amines and antibiotic susceptibility were addressed. METHODS AND RESULTS: The bacteriocin-producing Lact. sakei CRL1862 showed respectively bactericide and bacteriostatic effect against Listeria monocytogenes and Staphylococcus aureus in beaker sausage assay during 9 days of storage at 22 °C. The hydrolytic effect of Lact. sakei CRL1862 on protein extracts was evaluated by SDS-PAGE and reverse phase HPLC. A more pronounced proteolysis was evidenced in inoculated sarcoplasmic proteins compared with myofibrillar extracts with the generation of predominantly hydrophilic peptides and increase of total free amino acids concentration. Lactobacillus sakei CRL1862 produced neither histamine nor tyrosine and exhibited no resistance to the antibiotics assayed. CONCLUSIONS: Lactobacillus sakei CRL1862 effectively controlled the growth of L. monocytogenes and Staph. aureus; moreover, it was able to hydrolyse pork meat extracts generating peptides and amino acids, which may improve hygienic and sensorial attributes of fermented meat products. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of an integrated approach to evaluate the major traits of Lact. sakei CRL1862 showed it can be applied as an autochthonous functional starter in meat fermentation.


Subject(s)
Antibiosis , Food Microbiology , Lactobacillus/growth & development , Meat Products/microbiology , Amino Acids/analysis , Animals , Bacteriocins/biosynthesis , Biogenic Amines/biosynthesis , Drug Resistance, Bacterial , Fermentation , Hydrolysis , Lactobacillus/drug effects , Lactobacillus/metabolism , Listeria monocytogenes/growth & development , Muscle Proteins/metabolism , Staphylococcus aureus/growth & development , Swine
20.
Biochem J ; 433(1): 139-44, 2011 Jan 01.
Article in English | MEDLINE | ID: mdl-20950271

ABSTRACT

dcAdoMet (decarboxylated S-adenosylmethionine) is an essential intermediate in the synthesis of polyamines. Its content is normally very low, amounting to less than 5% of that of S-adenosylmethionine itself. It was found that in mice lacking spermine synthase there was a large increase in dcAdoMet and that overexpression of spermine synthase reduced the amount of this nucleoside. There was also an increase in dcAdoMet in cells derived from patients with Snyder-Robinson syndrome, a rare X-linked recessive human disease caused by SMS gene mutations that greatly reduce the content of spermine synthase. These results suggest that there is an inverse relationship between the amount of spermine synthase protein and the content of dcAdoMet and raise the possibility that some of the abnormalities seen in mammals deficient in spermine synthase might be due to changes in dcAdoMet pools.


Subject(s)
S-Adenosylmethionine/analogs & derivatives , Spermine Synthase/metabolism , Animals , Biogenic Amines/biosynthesis , Cells, Cultured , Decarboxylation , Humans , Mental Retardation, X-Linked/metabolism , Mice , Mice, Knockout , S-Adenosylmethionine/analysis , S-Adenosylmethionine/metabolism , Spermine Synthase/analysis
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