0 引言
【研究意义】赛里木镇位于新疆阿克苏地区拜城县,赛里木酸奶质地粘稠,拉出的长丝能达到1 m左右,又被称作赛里木拉丝酸奶 [1]。乳酸菌在胃肠道中繁衍增殖, 能够维持肠道菌群动态平衡、增强机体免疫力[2]。赛里木酸奶虽然具有良好的风味特征,但关于其中微生物组成的研究还不深入,传统的发酵方式该种酸奶的微生物组成复杂,组分及其相互作用也不明确。分离、保藏赛里木酸奶中的乳酸菌对于开发和利用优良菌种资源有着重要意义。【前人研究进展】玛依诺·木图拉[3]从赛里木酸奶中筛选得到得到2株产粘乳酸菌,分别为嗜热链球菌(Streptococcus thermophilic)和瑞士乳酸菌(Lactobacillus helveticus);高冬腊[4]在赛里木酸奶中分离得到1株德氏乳杆菌保加利亚亚种(Lactobacillus delbrueckii subsp.bulgaricus),均具有较好的益生特性。【本研究切入点】有关赛里木酸奶中乳酸菌的分离鉴定及特性分析文献较少,近些年来,对赛里木酸奶的相关研究在逐渐增加,但赛里木酸奶中微生物资源丰富,需进一步挖掘利用。【拟解决的关键问题】以采集到的12种赛里木酸奶为研究对象,筛选鉴定赛里木酸奶中分离得到的乳酸菌,检测生理生化特性。筛选益生效果好、食用安全的乳酸菌资源,有效扩大特色乳酸菌资源库和保护利用相关益生资源。
1 材料与方法
1.1 材料
1.1.1 原料
赛里木酸奶样品:分别采自新疆阿克苏地区拜城县赛里木镇周边。
供试菌株:枯草芽孢杆菌(Bacillus subtilis)、金黄色葡萄球菌(Staphylococcus aureus)、大肠埃希菌(Escherichia coli)、白色念珠菌(Candida albicans)、青霉菌(Penicillium)和黑曲霉(Aspergillus niger)菌株,均由新疆微生物资源保藏管理中心(Microbiological Culture Collection Center of Xinjiang,MCCCX)提供。
1.1.2 试剂
无水乙醇、冰乙酸、溴酚蓝、吐温-80、戈卢氏碘液、刚果红,均为分析纯,由北京鼎国昌盛生物技术有限责任公司提供。
吲哚试剂:取 0.1 g 对二甲氨基苯甲醛,用 5 mL 95%乙醇溶解,然后再缓慢加入 2 mL 浓盐酸。
α-萘胺试剂:1 g α-萘胺溶于 20 mL 蒸馏水和 150 mL 200 g/L 的乙酸溶液中。
对氨基苯磺酸试剂:0.5 g 对氨基苯磺酸溶于 150 mL 200 g/L 的乙酸溶液中。
抗生素滤纸片:青霉素 10 IU/片、红霉素 15 μg/片、四环素 30 μg/片、链霉素 10 μg/片、庆大霉素 10 μg/片、卡那霉素 30 μg/片、诺氟沙星 10 μg/片、氨苄西林 10 μg/片、利福平 5 μg/片、万古霉素 30 μg/片,购自北京天坛生物制品股份有限公司。
1.1.3 主要培养基
MRS 培养基、哥伦比亚血琼脂培养基和 LB 液体培养基,由青岛高科技工业园海博生物技术有限公司提供。
生物胺检测培养基:牛肉膏 5.0 g,胰蛋白胨 5.0 g,酵母粉 5.0 g,葡萄糖 0.5 g,柠檬酸铵 2.0 g,NaCl 2.5 g,K2HPO4·7H2O 2.0 g,吐温-80 1 mL,MgSO4 0.2 g,CaCO3 0.1 g,FeSO4 0.04 g,MnSO4 0.05 g,VB1 0.01 g,5-磷酸吡哆醛 0.05 g,溴甲酚紫 0.06 g,琼脂 20 g,蒸馏水 1 000 mL,pH 值 5.8~6.0。
改良氨基脱羧酶检测培养基:分别在生物胺检测培养基中加入 10 g/L 的对应前体氨基酸(精氨酸、酪氨酸、尸氨酸、色氨酸、组氨酸)。
硝基还原酶检测培养基:蛋白胨 10 g,硝酸钾 1 g,蒸馏水 1 000 mL,pH 值7.4 左右。
蛋白胨水培养基:蛋白胨 20 g,氯化钠 5 g,蒸馏水 1 000 mL,pH 7.4~7.6。
1.1.4 仪器与设备
超净工作台(苏州安泰空气技术有限公司);自动高压灭菌锅(日本 HIRAYAMA 公司);TS 恒温振荡器(德国耶拿分析仪器股份公司);PCR仪(赛默飞世尔科技(中国)有限公司);生化培养箱(上海福玛实验设备有限公司);立式冷藏柜(海尔 HAIER 公司);电热恒温水浴锅(上海齐欣科学仪器有限公司)。
1.2 方法
1.2.1 样品采集
赛里木酸奶样品采自新疆阿克苏地区拜城县,经灭菌的 50 mL 离心管分装,放入车载低温冰箱保存,采样结束后分别放入4℃和-80℃冰箱保存,备用。表1
表1 样品及来源
Tab.1
| 序号 Serial number | 编号 Code numble | 采集地点 Sampling position | 备注 Remark |
|---|---|---|---|
| 1 | H1 | 拜城县黑英山乡亚吐尔村4组 | 自产 |
| 2 | H2 | 拜城县黑英山乡8村1组 | 半商品化 |
| 3 | H3 | 拜城县黑英山乡8村2组 | 自产 |
| 4 | H4 | 拜城县黑英山乡明布拉克村大组 | 自产 |
| 5 | H5 | 拜城县黑英山乡1村4组 | 自产 |
| 6 | D1 | 拜城县赛里木酸奶厂 | 商品化 |
| 7 | M1 | 赛里木镇2村2大队 | 自产 |
| 8 | K1 | 拜城县克孜尔乡 | 自产 |
| 9 | S1 | 拜城县赛里木酸奶厂 | 商品化 |
| 10 | S4 | 赛里木镇托喀其买里村 | 半商品化 |
| 11 | S5 | 赛里木镇6村1组 | 自产 |
| 12 | S6 | 赛里木镇2村2大队 | 自产 |
1.2.2 菌株分离筛选
在无菌条件下吸1 mL 样品于9 mL的无菌生理盐水中, 振荡混匀, 采用梯度稀释法,将稀释液在含有1% CaCO3的MRS固体培养基上涂布,37℃倒置培养48 h。挑取有溶钙圈的细菌单菌落,进一步划线纯化后,转接至MRS斜面培养基上,37℃培养48 h, 4℃下保存备用。
1.2.3 菌株分子鉴定
挑取少量新鲜的乳酸菌单菌落,以菌株DNA为模板,使用16S rRNA通用引物27F (5'-AGAGTTTGATCCTGGCTCAG-3')和1 492R (5'-GGTTACCTTGTTACGACTT-3')进行PCR扩增,扩增产物在进行0.8%琼脂糖凝胶电泳检测之后送至生工生物工程(上海)股份有限公司进行测序分析。
将测序完成的DNA序列利用SeqMan (DNAStar 6.0)软件进行序列拼接,将得到的菌株的16S rRNA基因序列在EzTaxon数据库(http://www.ezbiocloud.net/)中比对分析,使用MEGA 7.0进行ClustalX多重比对,构建Neighbor-Joining系统发育树,确定菌株分类地位。
1.2.4 菌株的生长特性
以MRS液体培养基为基础培养基进行菌株的生长温度、耐盐、耐酸特性检验,在含0.5%猪胆盐的乳糖胆盐培养基上进行胆盐耐受性检验,牛奶胨化实验参考《常见细菌系统鉴定手册》[5]进行。
1.2.5 菌株的功能性酶活
将各乳酸菌株点接接种至相应功能酶筛选培养基上,30℃培养3~5 d,测定菌株的蛋白酶、果胶酶、酯化酶、淀粉酶和纤维素酶活性,其中,蛋白酶、果胶酶和酯化酶筛选直接观察透明圈;淀粉酶筛选用0.1%的碘液染色后观察透明圈;纤维素酶筛选用0.1%的刚果红染色后观察透明圈[6]。
1.2.6 菌株的抑菌活性
将指示菌株大肠埃希菌、枯草芽孢杆菌、金黄色葡萄球菌、白色念珠菌,接种于LB液体培养基中,30℃、120 ×g条件下振荡培养16 h。取100 μL发酵菌液,均匀涂布于MRS固体培养基。全部菌液平板吸收后,在设定区域的中央接种菌株,30℃培养48 h,确定出其产抑菌圈性能,充分的冲洗青霉菌、黑曲霉,根据要求将孢子悬液涂布后点接待测菌株,30℃环境条件下充分培养48 h后观察,判断出其产抑菌圈。
1.2.7 溶血性测定
将乳酸菌株接种到血琼脂平板上,在37℃下培养48 h,其附近产生透明的溶血环,对应为β-溶血;观察出现一定尺度的草绿色环,对应为α-溶血;未出现变化的,对应为γ-溶血。以金黄色葡萄球菌作为阳性对照。
1.2.8 有害代谢物质检测
1.2.9 菌株的自聚合能力测定
将乳酸菌接种至MRS液体培养基,37℃,120 ×g条件下培养48 h,将发酵后菌液在3 000 ×g、4℃条件下离心10 min,弃去上清液,保留菌体,重悬于无菌生理盐水中,调节600 nm下的OD值至0.5,记为A0h)。振荡混匀30 s,在酶标板中加入400 μL菌悬液,37℃条件下静置2 h。静置完成后,吸取200 μL上清液,测定其在,记为A2h,乳酸菌自凝率测定如下所示,取3次重复试验的平均值[10]。
自凝率(%)=(1-A2h/A0h)×100.
1.3 数据处理
每个样品测定 3 次取平均值,采用Excel作图。
2 结果与分析
2.1 酸奶中分离乳酸菌及其生理生化特性鉴定
研究表明,从赛里木酸奶中分离出4种乳酸菌菌分属于Eremococcus、Leuconostoc、Lentilactobacillus、 Streptococcus 4个属下的4个种。图1
图1
图1
赛里木酸奶中乳酸菌的系统进化树
Fig.1
Phylogenetic tree of lactic acid bacteria in Sayram Ketteki
2.2 菌株的生长特性
研究表明,其中4株乳酸菌在45℃正常生长,没有异常;所有菌株在生长过程中对5%的NaCl都有良好的耐受性,其中菌株S5-1能耐受7%的NaCl;pH 3.0时不生长,适当的增加酸碱度在pH为4时,开始微弱的生长。在含0.5%猪胆盐培养基上4株试验菌株的生长都没有受到影响。4株试验菌株的胨化能力均较好,生长过程中并不析出乳清;菌株H4-2发酵牛奶可产生浓郁香气,质地柔和细腻。表2
表2 菌株的生长情况
Tab.2
| 菌株 Strains | 温度 Temperature (℃) | NaCl(%) | pH | 胆盐 Bile salt (%) | 牛奶胨化 Milk peptonization | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| 20 | 45 | 3 | 5 | 7 | 3 | 4 | 0.5 | 胨化 Peptization | 产香 Fragrance | |
| S5-1 | + | + | + | + | + | - | w | ++ | + | + |
| D1-3 | + | + | + | + | - | - | w | ++ | + | + |
| H4-2 | + | + | + | + | - | - | w | ++ | + | ++ |
| H5-4 | + | + | + | + | - | - | w | ++ | + | + |
注:+:菌株正常生长;-:菌株没有生长;w:生长较弱
Note:+:Normal growth of strain; -:The strain did not grow; w:weak growth
2.3 菌株的自聚合能力测定
研究表明,4种菌株均表现出良好的自聚合能力,自凝率最高的为H4-2,达到83.46%,最低为S5-1,达到67.1%。 图2
图2
2.4 菌株的功能酶活性
研究表明,4株菌均具有蛋白酶活性,但不具有其他功能酶活性。表3
表3 菌株功能酶活性
Tab.3
| 菌株 Strains | 功能酶 Functional enzymes | ||||
|---|---|---|---|---|---|
| 蛋白酶 Protease | 淀粉酶 Amylase | 果胶酶 Pectase | 酯化酶 Esterase | 纤维素酶 Cellulase | |
| S5-1 | + | - | - | - | - |
| D1-3 | + | - | - | - | - |
| H4-2 | + | - | - | - | - |
| H5-4 | + | - | - | - | - |
2.5 菌株的抑菌活性
研究表明,4株菌均对大肠埃希菌有抑制作用,其中D1-3与H4-2的抑菌活性较强;4株菌对金黄色葡萄球菌有抑制作用,其中H4-2的抑菌活性较强;3株菌对枯草芽孢杆菌有抑制作用;仅有H4-2对白色念珠菌有抑制作用,所有菌均不抑制青霉菌和黑曲霉。表4
表4 菌株的抑菌活性
Tab.4
| 菌株 Strains | 病原指示菌 Pathogen indicator | |||||
|---|---|---|---|---|---|---|
| 大肠埃希菌 E.coli | 枯草芽孢杆菌 B.subtilis | 金黄色葡萄球菌 S.aureus | 青霉菌 Penicillium | 黑曲霉 Asp.niger | 白色念珠菌 C.albicans | |
| S5-1 | ++ | - | + | - | - | - |
| D1-3 | +++ | + | ++ | - | - | - |
| H4-2 | +++ | ++ | +++ | - | - | + |
| H5-4 | + | + | + | - | - | - |
2.6 菌株的生物安全性
研究表明,4株菌的生物安全性都达到较高水平,其中H5-4菌株在含L-色氨酸的平板上阳性,由此可判断出其产胺性能强,可转换前体形成色胺,其他的菌株都为阴性。在硝基还原酶检测结果表明,除H5-4菌株外都不产生硝基还原酶。所有菌株在吲哚实验中均为阴性结果,并且均不具有溶血性。表5
表5 菌株的有害代谢物质检测
Tab.5
| 菌株 Strains | 生物胺 Biogenic amines | 吲哚 Indoles | 溶血性 hemolytic | 硝基还原酶 Nitroreductase | ||||
|---|---|---|---|---|---|---|---|---|
| 精胺 Spermine | 酪胺 Tyramine | 色胺 Tryptamine | 尸胺 Cadaverine | 组胺 Histamine | ||||
| S5-1 | - | - | - | - | - | - | - | - |
| D1-3 | - | - | - | - | - | - | - | - |
| H4-2 | - | - | - | - | - | - | - | - |
| H5-4 | - | - | + | - | - | - | - | + |
| *E.coli | (CK: -) | + | + | + | ||||
3 讨论
3.2 赛里木酸奶独特风味的形成与其复杂的微生物组成有着直接的关系,高冬腊[4]在赛里木酸奶中利用高通量测序发现,酸奶中的优势菌门为厚壁菌门,优势细菌属为乳杆菌属和链球菌属。王子涵[14]利用高通量测序技术在赛里木酸奶中得到的结果基本相同,酸奶中链球菌属为优势菌属,其次为乳杆菌属。研究在阿克苏地区赛里木镇周边收集到12份赛里木酸奶样品,分离筛选得到36株乳酸菌,经16S rRNA基因序列分析鉴定为Eremococcus、Leuconostoc、Lentilactobacillus、Streptococcus 4个属下的4个种,分别为S5-1(Eremococcus coleocola)、H4-2(Lentilactobacillus kefiri)、H5-4(Leuconostoc mesenteroides subsp.mesenteroides)及D1-3(Streptococcus thermophilus)。
3.4 乳酸菌的抗菌能力,是其众多益生功能中的一个重要特性,能够避免胃肠道感染的出现[17]。王融等[18]在研究鱼类肠道乳酸菌时发现,肠道乳酸菌在生长过程中会分泌出胞外物质,这种物质可以将病原菌与胃肠道黏膜隔绝开来,此外肠道乳酸菌还会与病原菌竞争营养物质,同时分泌出具有抗菌活性的代谢产物,进而达到抑制病原菌生长繁殖的效果。蔡静静等[19]在新疆伊犁地区乳制品中分离得到7株对大肠埃希氏菌具有较好抑制性的乳酸菌。研究中4株乳酸菌均对大肠埃希菌及金黄色葡萄球菌均有抑制作用,其中H4-2的抑菌活性较强;3株菌对枯草芽孢杆菌有抑制作用;仅有H4-2对白色念珠菌有抑制作用。不同乳酸菌之间抑菌范围及抑菌效果均存在较大差异,可能是由于不同乳酸菌所产生的的代谢产物不同,仍需进一步研究。
4 结论
在12种赛里木酸奶样品中筛选得到4种乳酸菌,分属于Eremococcus、Leuconostoc、Lentilactobacillus、 Streptococcus 4个属下的4个种,试验菌株可耐受0.5%胆盐,0.5%NaCl,且具有较好耐高温、牛奶胨化和抑制细菌等特性,并展现出良好的自聚合能力。4株乳酸菌均不溶血,且吲哚检测试验均呈阴性,除H5-4外均不产生硝基还原酶及生物胺。
参考文献
Some current applications, limitations and future perspectives of Lactic acid bacteria as probiotics
[J].
库车小白杏内生细菌多样性及其产酶特性
[J].
Diversity and functional enzymes of endophytic bacteria in Kuqa Apricot
[J].
This study aimed to analyze the diversity of endophytic bacteria in postharvest Kuqa apricot for the purpose of discovering potential microbial resources and their functions. The composition of the endophytic bacterial community was analyzed by high-throughput sequencing. Meanwhile, endophytic bacteria were isolated using the traditional culture method, and identified by 16S rRNA gene sequence analysis. The enzyme activities of the isolates were tested. High-throughput sequencing results showed that 406 operational taxonomic units (OTUs) affiliated to 94 genera of 9 phyla were observed. Bacteroidetes, Bacillus, Paenibacillus and unclassified bacteria were the most abundant species. A total of 99 isolates were obtained, which were classified into 28 genera in the phyla of Actinobacteria, Proteobacteria, Firmicutes and Bacteroidetes. In addition, seven genera not detected by high-throughput sequencing and 5 potential novel species were discovered. At the same time, several isolates were found to be capable of producing protease, lipase, cellulase, amylase and pectase, which could provide abundant materials for further mining and development of potential microbial resource and enzymes from Kuqa apricot. The results provide a new idea for studying microbial deterioration of Kuqa apricot.
Raw donkey milk as a source of enterococcus diversity: assessment of their technological properties and safety characteristics
[J].
一株乳酸乳球菌产γ-氨基丁酸能力及其安全性评价
[J].
γ- aminobutyric acid-producing ability and safety evaluation of Lactococcus lactis
[J].
乳酸菌体外黏附性方法的评价及相关性研究
[J].
Correlation between Methods for Evaluating the Adhesion of Lactic Acid Bacteria
[J].
The influence of the wooden equipment employed for cheese manufacture on the characteristics of a traditional stretched cheese during ripening
[J].
Diversity of yeast species during fermentative process contributing to Chinese Maotai-flavour liquor making
[J].
中国传统发酵食品微生物多样性及其代谢研究进展
[J].
Research progress of microganism and its metabolism in traditional fermentation food
[J].
赛里木酸奶细菌多样性及挥发性风味物质成分分析
[J/OL].
Analysis of bacterial diversity and volatile flavor compounds in sayram yogurt
[J/OL].
Lactic acid bacteria diversity of African raw and fermented camel milk products reveals a highly competitive, potentially health-threatening predominant microflora
[J].
In vitro evaluation of probiotic potential of Lactobacillus cultures of human origin capable of selenium bioaccumulation
[J].
Isolation of lactic acid bacteria showing antioxidative and probiotic activities from klimchi and infant fece
[J].
鱼类肠道乳酸菌的抑菌特性及机理研究
[J].
Study on antibacterial characteristics and mechanism of lactic acid bacteria in fish intestine
[J].
新疆伊犁地区乳制品中乳酸菌发酵和益生特性及其复合发酵方案优化
[J].
Fermentation and probiotic properties of lactic acid bacteria isolated from dairy products in Yili, Xinjiang and optimization of their mixtures for mixed-culture fermentation
[J].The acidification ability and aroma-producing ability of seven strains of lactic acid bacteria preserved in our laboratory for yogurt fermentation and the texture properties of yogurt made with each of the strains were measured. The probiotic characteristics of these strains were evaluated by their antibacterial properties, gastrointestinal juice tolerance and antibiotic susceptibility. Strains with better performance were selected and combined to obtain the optimal mixed starter culture. The results of single-strain fermentation showed that the coagulation time of yogurts made separately with Leuconostoc lactis XKN1-5 and Lactobacillus delbrueckii subsp. bulgaricus BSTS6-4 was shorter (about 7 h), and the acidity reached around 96 °T; their acidification abilities were better than those of other strains. The yogurts fermented separately by L. delbrueckii subsp. bulgaricus BSTS6-3 and Streptococcus thermophilus XSXN1-2 had excellent texture characteristics. L. delbrueckii subsp. bulgaricus BSTS6-1 and S. thermophilus XSXN1-1 had the best aroma-producing ability and the yogurt fermented by strain XSXN1-1 contained 2.02 mg/L of diacetyl and 16 mg/L of acetaldehyde, indicating significantly improved flavor. The results of probiotic properties showed that L. fermentum BST2-3 and L. delbrueckii subsp. bulgaricus (BSTS6-1, BSTS6-3, and BSTS6-4) could inhibit the growth of pathogenic bacteria, and Lactobacillus fermentum BST2-3, L. delbrueckii subsp. bulgaricus BSTS6-1, BSTS6-4) had greater adhesion ability; the number of viable L. fermentum BST2-3 was 106 CFU/mL after 9 h of simulated gastrointestinal juice treatment. All the seven strains showed sensitivity to most tested antibiotics and were of high safety. Based on the ability to produce acids and aroma compounds, yogurt texture and probiotic properties, the optimal strain combination for mixed-culture fermentation was obtained as L. delbrueckii subsp. bulgaricus (BSTS6-4 and BSTS6-3) + S. thermophilus XSXN1-1 + L. fermentum BST2-3.
强化发酵对辣椒中生物胺及理化指标影响的研究
[J].
Effect of Intensified Fermentation on Biogenic Amines and Physicochemical Indexes of Chili
[J].
Antioxidative Effect of Biogenic Amine on the Peroxidation of Linoleic Acid
[J].
Focused Review: Agmatine in Fermented Foods
[J].
发酵肉制品加工中衍生的非健康因子控制研究进展
[J].
Recent progress on control of the unhealthy factors deriving from spontaneously fermented meat products processing
[J].
Biogenic amine formation and bacterial contribution in Cheonggukjang, a Korean traditional fermented soybean food
[J].
腌干鱼贮藏过程生物胺的变化及其货架期研究
[J].为建立以生物胺变化预测腌干鱼在贮藏过程中品质和安全食用货架期的回归方程,以多脂的腌干带鱼和低脂的腌干金线鱼为原料,研究其在常温贮藏过程中的理化指标、主要微生物和生物胺的变化及相关关系,并应用逐步回归法进行回归分析,初步建立分析腌干鱼制品的货架期预测模型。结果表明,腌干带鱼和金线鱼在贮藏过程中pH、水分含量和Aw不断下降。葡萄球菌及微球菌、乳酸菌、菌落总数在贮藏3周内呈上升趋势,从第4周开始逐渐下降,而假单胞杆菌和肠杆菌在贮藏第4周之后未检出;组胺、尸胺和腐胺为腌干鱼贮藏过程主要胺类物质,呈上升趋势。5种生物胺(组胺、尸胺、腐胺、色胺、酪胺)、总生物胺量、生物胺相关指标、理化指标、假单胞杆菌均与贮藏时间高度相关,微生物中只有假单胞杆菌与生物胺及其相关指标有相关性;生物胺与理化指标也呈高度负相关。本研究结果为腌干鱼制品货架期体系的建立提供了参考依据。
Study on change of biogenic amines and its shelf life of dried-salted fish at storage
[J].To establish the regression equation to predict the quality and safety dried-salted fish on storage depend on the change of biogenic amines, the change rule of biogenic amines of dried-salted fish during storage at room temperature and predictive modeling of shelf life, the changes and mutual relationship of physicochemical indexes, main microorganism and biogenic amines in dried-salted fatty hairtail and low fat threadfin bream during storage were studied. The preliminary establishment of dried-salted fish shelf life prediction model was used stepwise regression analysis method. The results showed that pH, water content and Aw in dried-salted hairtail and threadfin bream continued to decline during storage, total plate counts, lactic acid bacteria, <em>micrococcus</em> and <em>staphylococcus</em> contents were on the rise in three weeks, then decreased beginning from fourth week, while <em>pseudomonas</em> and <em>enterobacterium</em> contents were not checked out after the fourth week. Major biogenic amines, such as histamine, cadaverine and putrescine, were on the rise during storage. Five kinds of biogenic amines (histamine, cadaverine, putrescine, tryptamine, tyramine), total biogenic amine, relevant biogenic amine indices, physicochemical indicators and <em>pseudomonas</em> had a significant correlation with the storage time. Only <em>pseudomonas</em> was related to the biogenic amine and its related indicators. It is to provide a reference foundation for establish the system of shelf life about dried-salted fish.
Tyramine and histamine risk assessment related to consumption of dry fermented sausages by the Spanish population
[J].
