Biogenic amine production by Oenococcus oeni during malolactic fermentation of wines obtained using different strains of Saccharomyces cerevisiae

Twenty-six wild Oenococcus oeni strains were investigated for their ability to form biogenic amines during malolactic fermentation in synthetic medium and in wine. Eight strains produced histamine and tyramine in screening broth at concentrations of 2.6-5.6 mg/L and 1.2-5.3 mg/L, respectively. Based on their ability to form biogenic amines, five strains were selected to inoculate three wines obtained by the fermentation of three different Saccharomyces cerevisiae strains (A, B, and C). All bacterial strains could perform malolactic fermentation for short periods in wine C, whereas only one strain performed complete malolactic fermentation in wines A and B. Two O. oeni strains (261 and 351) produced histamine and tyramine in wine C. Time-course analysis of these compounds showed that for both strains, histamine and tyramine production began at day 10 and finished on day 25, after the end of malolactic fermentation. These results indicate that the ability of O. oeni to produce histamine and tyramine is dependent on the bacterial strain and on the wine composition, which in turn depends on the yeast strain used for fermentation, and on the length of bacteria-yeast contact time after the completion of malolactic fermentation.     

Effect of γ‐aminobutyric acid supplementation on the composition of Chinese rice wine

This study investigated the effect of γ‐aminobutyric acid (GABA) supplementation on the composition of Chinese rice wine. GABA was added at 188.5 and 377 mg/L at the beginning of primary fermentation or post‐fermentation. Addition of GABA during primary fermentation had no impact on alcohol and total acid formation or the utilisation of sugar. However, total free amino acids, total essential amino acids and GABA in rice wine supplemented with GABA were significantly lower than the control rice wine without supplementation. Alcohol formation, total acid, sugar content and total essential amino acids were similar compared with the control. However, the GABA post‐supplementation was significantly higher, suggesting that GABA supplementation post‐fermentation process could enhance the GABA content in rice wine. © 2018 The Institute of Brewing & Distilling     

BIOGENIC AMINES IN FINNISH DRY SAUSAGES

A study was conducted to determine the biogenic amine levels of Finnish dry sausages and to compare the results with those of other surveys of different types of fermented sausages. Sausages with high amine concentration were analyzed microbiologically in order to identify amine-producing microorganisms. Tyramine was the most common amine: the mean of 68 samples was 82 mg/kg and the mode 110 mg/kg. Other vasoactive amines found were histamine (<1–200 mg/kg), phenylethylamine (<1–48 mg/kg) and tryptamine (<10–91 mg/kg). Increased amounts of the spoilage indicator amines putrescine and cadaverine were found from most of the samples containing vasoactive amines. The formation of amines can be linked with problem(s) in hygienic conditions of raw materials and manufacturing practices and it is thus proposed that the sum of tyramine, histamine, putrescine and cadaverine could serve as an indicator of quality in fermented meat products.     

Effect of fermentation with mixed starter cultures on biogenic amines in bighead carp surimi

The effects of one group mixed starter cultures, combined with Lactobacillus casei, Streptococcus lactis, Saccharomyces cerevisiae Hansen, and Monascus anka, and a batch without starter as control on biogenic amines accumulation in the bighead carp surimi during fermentation were investigated. Determination of five different biogenic amines was carried out by reverse‐phase high‐performance liquid chromatography with UV detection. Results showed that mixed starter cultures decreased the pH quickly from initial pH value of 6.4 to 5.2, inhibited the growth of contaminant microorganisms, such as Enterobacteriaceae and Pseudomonas, present in the raw materials, and suppressed the accumulation of histamine, tyramines, spermine and spermidine. Fermentation inoculated with combination of Lactobacillus casei, Streptococcus lactis, Saccharomyces cerevisiae Hansen, and Monascus anka with negative‐decarboxylase activity may prevent biogenic amine formation in the fermented ssurimi and improve hygienic quality of fermented surimi.     

利用高效液相色谱法测定不同发酵时长郫县豆瓣酱中的8种生物胺

本研究基于高效液相色谱(HPLC)技术,测定了不同发酵时长(1~96个月)郫县豆瓣酱中色胺、β-苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺和精胺8种生物胺的含量。结果显示,HPLC分析中,8种生物胺的检出限在0.17~0.35 mg/L之间,定量限在0.55~0.92 mg/L之间,各生物胺标品在1~60 mg/L的质量浓度范围内线性关系良好,可被准确定量;各生物胺回收率在73.10%~104.13%之间,说明本方法具有较高的重复性和灵敏度。腐胺、尸胺、酪胺和组胺是郫县豆瓣酱中的主要生物胺;随着发酵时间的延长,腐胺含量快速上升,尸胺含量在发酵前54个月迅速上升,之后明显下降,酪胺和组胺在发酵54个月后含量有所增加;其他生物胺含量较低,且发酵期间变化不明显;郫县豆瓣酱中总生物胺含量随发酵时间的延长不断积累,从(37.86±6.69) mg/kg增加到(155.09±8.53) mg/kg,其含量在发酵前54个月快速上升,之后趋于稳定。目前食品中的多数生物胺尚没有限量标准,但参考现有的毒理学数据来看,本研究中郫县豆瓣酱的生物胺含量水平相对安全。     

Characterization by Volatile Compounds of Microbial Deep Spoilage in Iberian Dry-Cured Ham

Abstract: In the present study, volatile compounds of spoiled dry-cured Iberian ham with deep spoilage or “bone taint” were analyzed and correlated with level of spoilage and the microorganisms detected. Volatile compounds extracted by a solid phase micro-extraction technique were assayed by gas chromatography/mass spectrometry. The spoiled hams were evaluated sensorially, and the correlations among volatile compounds, spoilage level, and microbial counts were studied. The spoiled hams had higher concentrations of hydrocarbons, alcohols, acids, esters, pyrazines, sulfur compounds, and other minor volatile compounds than unspoiled hams. The sensorial analysis showed that the spoilage level of hams correlated with several volatile compounds, most of them associated with Gram-positive catalase positive cocci and Enterobacteriaceae counts. Cyclic compounds such as cyclohexanone, some ethers, and pyrazines should be considered as indicators to monitor incipient microbial deep spoilage in the elaboration of this meat product.     

新疆赤霞珠葡萄酒发酵过程中生物胺含量的变化

利用高效液相色谱技术分析葡萄酒发酵过程中8种生物胺含量的变化。结果表明,葡萄酒发酵过程中8种生物胺总含量整体呈上升趋势。色胺含量最高,最高可达到98.03 mg/L;腐胺和组胺含量较低,含量均不超过2.48 mg/L;精胺和亚精胺含量变化相对平稳,波动范围在1.81～3.07 mg/L之间,除色胺外,其他生物胺含量均不超过4.50 mg/L。苯乙胺、尸胺、酪胺等在酒精发酵后期和苹果酸-乳酸发酵初期大幅上升,在苹-乳发酵后期,酪胺和亚精胺含量有所下降,其他生物胺含量均有一定程度上升。     

Histamine and Tyramine during Storage and Spoilage of Anchovie, Engraulis encrasicholus: Relationships with Other Fish Spoilage Indicators

The formation of biogenic amines (histamine and tyramine) during Engraulis encrasicholus storage and spoilage was studied and compared with changes of other fish spoilage indicators: trimethylamine nitrogen, volatile basic nitrogen, hypoxanthine and pH. Good correlations were observed between biogenic amines and the other indicators, at both refrigeration (4–6°C) and room temperature (18–22°C).Two trials were carried out at each temperature. Differences were observed in levels of biogenic amines as well as other indicators in experiments carried out under similar conditions.     

Investigation of mashing regimes for low‐alcohol beer production

Low‐alcohol beer can be obtained by physical and biological methods. The group of biological methods includes modification of the mashing regimes and changes in the fermentation process. The aim of the present work was to study two mashing regimes for low‐alcohol beer production. The increase in the mashing duration at 50 °C led to a linear increase in the extract and the concentration of reducing and fermentable sugars in the wort. It was found that the rate of formation of reducing sugars was higher than that of the formation of fermentable sugars, which can be used for the optimization of the mashing process. The introduction of a pause at 77 °C did not lead to a substantial increase in the concentration of fermentable extract, but did lead to an increase in the total and non‐fermentable extract. The available nitrogen content in the laboratory wort was in the range of 120–150 mg/dm³. As a result of conducting fermentation processes with the top‐fermenting yeast strain Saccharomyces cerevisiae S‐33, it was found that the combination of a small amount of fermentable sugars and a low fermentation temperature led to a beer being obtained that met the requirements for a low‐alcohol beverage. Copyright © 2016 The Institute of Brewing & Distilling     

Microbiological attributes and biogenic amine content of probiotic Turkish fermented sausage

Microbiological attributes and biogenic amine content of Turkish fermented sausage manufactured by using probiotic starter cultures (Lactobacillus casei, L. acidophilus or their combination) were investigated before and after fermentation-drying period and during refrigerated storage at 4 ± 1 °C for 8 months at 2 month intervals. As results of the study, during fermentation and storage biogenic amine content (histamine, putrescine, cadaverine and tyramine) of the samples were increased significantly. Salmonella, Staphylococcus aureus, coliform and fecal coliform microorganisms were not detected during fermentation and storage. Probiotic microorganism counts of all samples were higher than the lower limit of 6.0 log cfu/g which is requested for probiotic foods.     

Biodiversity and safety aspects of yeast strains characterized from vineyards and spontaneous fermentations in the Apulia Region,Italy

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.     

GROWTH OF AEROBIC WILD YEASTS

Wild yeasts of the genera Debaryomyces, Hansenula and Pichia are commonly considered to be associated with spoilage only under aerobic conditions. However, in pure cultures in either wort or a synthetic medium of yeast nitrogen base + 10% glucose, yeasts of these genera grew as well as a brewing strain of Saccharomyces cerevisiae under anaerobic conditions. Growth of S. cerevisiae was increased by the addition of unsaturated fatty acid (Tween 80) or ergosterol to the medium for anaerobic culture. No equivalent requirement was observed for the wild yeasts examined. Indeed, growth of the wild yeasts was often reduced by the addition of Tween 80, which as a surfactant prevented formation of the surface film of growth. Even under anaerobic conditions, these yeasts grew best with a surface pellicle. Although capable of good anaerobic growth in pure culture, growth of the wild yeasts was suppressed under anaerobic conditions in mixed culture with S. cerevisiae, simulating a contaminated brewery fermentation. However, the contaminants competed successfully with S. cerevisiae under aerobic conditions. There was no evidence of a “killer” effect, but prevention of pellicle formation, or production of inhibitory levels of pH or ethanol under anaerobic conditions could explain the suppression of wild yeasts under anaerobic fermentation conditions.     

Levels of biogenic amines as a measure of the quality of the beer fermentation process: Data from Belgian samples

Biogenic amines were determined by HPLC in four beer types. In spontaneously fermented beers (SF beers), the amounts of vasoactive amines reached high levels, namely a mean value (1) above 20 mg/l for tyramine and (2) close to 10 mg/l for histamine. Considering the bacterial origin of these amines, we established a calculation formula for a Beer biogenic amine index (Beer BAI), reflecting the microbiological quality of the fermentation process. Using this formula we determined a mean Beer BAI value lying between 0.84 ± 0.89 (high quality) in low fermented beers and 16.2 ± 13.9 in SF beers. BAI values ⩾10 (poor hygienic fermentation process) corresponded to beers showing values of vasoactive BA (>10 mg/l) that could cause health troubles in certain types of consumers (people under treatment with monoamine-oxidase medication or genetically more sensitive to food-born BA).     

West African sorghum beer fermented with Lactobacillus delbrueckii and Saccharomyces cerevisiae: shelf‐life and consumer acceptance

The fermentation profiles, shelf‐life and consumer acceptance of traditional West African sour sorghum beer (pito) fermented with pure commercial starter cultures of lactic acid bacteria (L. delbrueckii) and yeast (Saccharomyces cerevisiae) were evaluated. The beers from this ‘pure culture’ approach were compared with the spontaneous fermentation of pito wort. Lactic acid formation, pH change and extract utilisation were monitored during fermentation. Lactic acid content was used as a measure of sourness to establish the spoilage level of over‐sourness. Further, regression models relating sourness to the time the drink was kept were used to predict the shelf‐life. Consumer acceptance of the product was evaluated using a novel nine‐point hedonic scale. The pure culture and traditionally fermented beers followed similar lactic acid and fermentation profiles but strain‐specific differences were observed. Similar levels of pH, lactic acid level and extract utilisation were achieved. An improvement in shelf‐life of two days was found over traditionally fermented pito. There was no statistical difference between the two pito products for overall liking and taste. However, there was a preference for the aroma of the pure culture pito. It is suggested that the use of pure cultures will facilitate the scale‐up of pito production. © 2019 The Institute of Brewing & Distilling     

高效液相法测定草鱼糜发酵过程中生物胺变化

本文采用纳豆芽孢杆菌和戊糖片球菌组成的混合发酵剂对草鱼鱼糜进行发酵,建立了发酵鱼制品中六种生物胺(色胺、腐胺、尸胺、组胺、酪胺、亚精胺)的高效液相色谱分析方法,研究了鱼糜发酵过程中生物胺含量的变化以及微生物数量的变化。结果发现,在30 min内各种生物胺得到很好的分离,且线性关系良好(R2>0.999),回收率在97.35%~103.00%,精密度、重现性RSD均小于10%。添加混合发酵剂可以明显抑制金黄色葡萄球菌、假单胞菌和大肠杆菌等杂菌的生长,降低发酵过程中腐胺、尸胺、组胺和酪胺含量的积累;利用纳豆芽孢杆菌,戊糖片球菌组成的混合发酵剂以及一系列负控脱羧酶活动能有效抑制鱼糜发酵过程中生物胺的形成,抑制腐败菌和致病菌的生长,提高鱼糜发酵食品的品质。      

Formation of biogenic amines throughout the industrial manufacture of red wine

Changes in biogenic amines (histamine, methylamine, ethylamine, tyramine, phenylethylamine, putrescine, and cadaverine) were monitored during the industrial manufacture of 55 batches of red wine. The origin of these amines in relation to must, alcoholic fermentation, malolactic fermentation, sulfur dioxide addition, and wine aging and the interactions between amines and their corresponding amino acids and pH were statistically evaluated in samples from the same batches throughout the elaboration process. Some amines can be produced in the grape or the musts (e.g., putrescine, cadaverine, and phenylethylamine) or can be formed by yeast during alcoholic fermentation (e.g., ethylamine and phenylethylamine), although quantitatively only very low concentrations are reached in these stages (less than 3 mg/liter). Malolactic fermentation was the main mechanism of biogenic amine formation, especially of histamine, tyramine, and putrescine. During this stage, the increase in these amines was accompanied by a significant decline in their amino acid precursors. Significant correlations between biogenic amine formation and the disappearance of their corresponding amino acids were observed, which clearly supports the hypothesis that malolactic bacteria are responsible for accumulation of these amines in wines. No increase in the concentration of biogenic amines was observed after SO2 addition and during wine aging, indicating that sulfur dioxide prevents amine formation in subsequent stages.     

HOP SUBSTANCES AND YEAST BEHAVIOUR

Top fermentation yeast strains vary in their ability to form a yeast head in the absence of hop substances. However, the addition of isohumulone or humulone to unhopped wort stimulates yeast-head formation. A quantitative method for the estimation of hop substances bound to the yeast cell is described and is used to demonstrate that in top fermentation, most of the adsorbed bitter substances are present in the head. It is shown that most of the hop substances in the yeast head are transported there by bubbles of carbon dioxide; in bottom fermentation, considerable quantities are present in the surface foam. In comparison with hopped wort the fermentation of unhopped wort with Saccharomyces cerevisiae (N.C.Y.C. 240) is characterized by an increased amount of yeast in suspension during fermentation, although there is no increase in the rate of attenuation.     

Production of Biogenic Amines and Their Potential Use as Quality Control Indices for Hake (Merluccius merluccius, L.) Stored in Ice

ABSTRACT: The production of tyramine, histamine, putrescine, cadaverine, agmatine, and spermidine was studied for use as quality indices in muscle of gutted hake stored in ice for 25 d. The changes in biogenic amine concentrations were correlated with trimethylamine nitrogen levels. All concentrations except for spermidine, increased progressively throughout storage. During storage, cadaverine reached 72.14 mg/kg while there were no significant increases in histamine and tyramine. The levels of free amino acids, which may be metabolized into biogenic amines, were not correlated to respective biogenic amines levels. The production of cadaverine and agmatine occurred prior to spoilage and these biogenic amines may indicate freshness of hake stored in ice.     

Application of the simplex method for optimization of chromatographic analysis of biogenic amines in fish

The aim of this study was to optimize the separation of biogenic amines with ion-exchange high-performance liquid chromatography, using the modified simplex method. A modified chromatographic response function was used, and two methodological parameters were selected as simplex variables: the proportion of the organic solvent 2-propanol in the mobile phases, added as a new mobile phase, and the variation of the gradient curve at one specific step in the elution program. In optimum conditions the correct resolution and separation of the biogenic amines (tyramine, β-phenylethylamine, histamine, tryptamine, putrescine, cadaverine, agmatine, spermidine and spermine) were completed in 16 min, reducing the analysis time by 9 min compared with the original method. The regression coefficients were higher than 0.99 in all the amines, except for spermine (0.989). The detection limits varied between 0.06 mg/L, for histamine and agmatine, and 0.22 mg/L, for tryptamine, while retention time repeatability, determined as a relative standard deviation, was between 4.64% for histamine and 11.95% for tryptamine. The repeatability variation for the amine concentrates was found between 0.71% in cadaverine and 3.23% for tryptamine. Applying the optimized method to samples of refrigerated sardine and sea bream demonstrated the effectiveness of the method in these fish species.     

Variation in content of tyramine,histamine, 2-phenylethylamine,tryptamine and their precursor amino acids in a Chardonnay wine by using different commercial active dry yeasts and nitrogen sources

Wine is a fermented beverage that could be affected by high concentrations of biogenic amines, thus altering organoleptic and health properties. In this work, the capacity of different selected yeast strains regarding to affect the content of tyramine, histamine, 2-phenylethylamine, tryptamine and their precursor amino acids (pAA) during fermentation has been described. Overall, biogenic amines (BAs) and pAA contents appeared significantly related both to the strain and to the nutrient supplementation applied. Among BAs, without and (with) nitrogen source addition, the concentrations covered a range between 0.1 and 9.5 (0.1–10.3) mg L⁻¹ for tyramine; 0.1 and 4.4 (0.1–4.6) mg L⁻¹ for histamine; 0.4 and 1.1 (0.4–1.4) mg L⁻¹ for 2-phenylethylamine; and 0.02 and 0.14 (0.01–0.12) mg L⁻¹ for tryptamine, respectively. Considering that also wine yeast species are responsible for BAs formation, the evidence from fermentation trials underlined the relevance of a careful choice of ADY strains and nitrogen sources in the management of alcoholic fermentation in wine.