四川区域自然发酵香肠及人工接种牦牛肉发酵香肠中生物胺含量的研究

以四川省10 区域自然发酵香肠及人工接种发酵剂的牦牛肉香肠共21 个样品为研究对象,测定发酵香肠中的生物胺含量。结果发现,21 个发酵香肠样品中均检测到酪胺、亚精胺、精胺、尸胺、腐胺、色胺及组胺,β-苯乙胺均未检出;生物胺总量在57.34～411.12 mg/kg,除采自凉山州西昌的自然发酵牦牛肉香肠和广安的自然发酵猪肉香肠中酪胺含量超过了美国食品及药品管理局（Food and Drug Administration,FDA）规定标准（酪胺含量≤100 mg/kg）,其余均不存在生物胺安全问题。     

传统自然发酵四川香肠加工贮藏过程中生物胺含量变化

采用高效液相色谱法对传统自然发酵四川香肠加工贮藏过程中生物胺(色胺、苯乙胺、腐胺、尸胺、组胺、酪胺和亚精胺)含量变化进行测定。结果表明:传统自然发酵四川香肠原料肉中只检测出了色胺、苯乙胺和亚精胺,贮藏120天检测出了7种生物胺,除亚精胺外,6种生物胺(色胺、苯乙胺、腐胺、尸胺、组胺和酪胺)的含量在加工过程中均显著增加(P0.01)。传统自然发酵四川香肠中以酪胺、组胺、尸胺和腐胺为主,含量分别达到153.06、127.90、132.46和92.83 mg/kg,4种生物胺占生物胺总量的89.66%。传统自然发酵四川香肠中生物胺(色胺、苯乙胺、组胺和酪胺)含量均低于20 mg/100 g。生物胺总量在加工贮藏过程中变化显著(P0.01),总量低于1 000 mg/kg。     

混合发酵剂对川味香肠发酵成熟过程中生物胺含量变化的影响

以植物乳杆菌、戊糖片球菌和葡萄球菌作为发酵剂接种至川味香肠,采用高效液相色谱法测定川味香肠发酵成熟过程中生物胺含量的变化。结果表明:成熟结束时,除苯乙胺外,A组(接种组)中的色胺、腐胺、尸胺、组胺、酪胺和亚精胺的含量显著低于B组(自然发酵组)(P0.01),说明混合发酵剂能显著抑制川味香肠发酵成熟过程中生物胺的形成(P0.01)。A组香肠在发酵成熟过程中除酪胺含量最高达到103.05 mg/kg外,其余生物胺含量均低于100 mg/kg;B组香肠在发酵成熟过程中腐胺、尸胺、组胺和酪胺含量超过100 mg/kg,因此接种混合发酵剂可以更好地保障川味香肠的安全性。     

混合菌种发酵生产低酸度川味香肠加工过程中生物胺含量的变化

接种混合发酵剂并在控温控湿条件下生产低酸度川味香肠,对加工过程中生物胺(色胺、苯乙胺、腐胺、尸胺、组胺、酪胺和亚精胺)含量变化进行研究。结果表明:干燥结束时,低酸度川味香肠中除亚精胺以外,其他生物胺含量均低于人工控温控湿不接种组(B组)、自然条件接种组(C组)、自然条件不接种组(D组),其中色胺、苯乙胺、腐胺、组胺和酪胺的含量均显著低于未接种组(P0.01),说明混合发酵剂在适宜的温湿度条件下能明显抑制低酸度川味香肠中色胺、苯乙胺、腐胺、组胺和酪胺积累(P0.01);混合发酵剂对低酸度川味香肠中尸胺的抑制作用不显著(P0.05)。     

休闲发酵色拉米中生物胺的测定与分析

人体摄入过多的生物胺时会出现许多不良反应，本实验对乳杆菌（米酒乳杆菌和戊糖乳杆菌）和片球菌（戊糖片球菌和小片球菌）在休闲发酵色拉米中生物胺的生成量进行了测定与分析。结果表明：供试乳杆菌和片球菌均能产生不等量的组胺、尸胺、腐胺、酪胺、亚精胺和精胺，但都不产生色胺，产品中的生物胺生成量表现出明显的菌株效应。采用复配菌种（片球菌＋葡萄球菌）发酵可以明显降低香肠中组胺、尸胺、腐胺和酪胺以及总生物胺的含量。     

传统中式香肠成熟过程中生物胺的生物控制

以传统中式香肠成熟过程中生物胺含量的变化为研究对象,重点介绍了色胺、腐胺、尸胺、组胺、酪胺的变化。在香肠加工过程中添加发酵剂和复合植物提取物来控制香肠中生物胺的积累,利用高效液相色谱仪检测成熟过程中生物胺的变化。结果表明,添加发酵剂可有效地抑制色胺、腐胺、尸胺、酪胺的生成;复合植物提取物只是在抑制酪胺含量的增加上有显著效果;同时加入发酵剂和复合植物提取物对色胺、尸胺、组胺和酪胺的抑制效果更显著,特别是组胺,在生产结束时含量为0。发酵剂和复合植物提取物对精胺和亚精胺基本没有影响。     

豇豆泡菜中产生物胺菌株的筛选鉴定及其产胺特性研究

传统四川泡菜在发酵过程中由于部分微生物代谢作用而产生的生物胺对人体有毒害作用，为探究泡菜中主要产胺微生物种类及其产胺特性，以自然发酵和卤水发酵豇豆泡菜为原料，在不同发酵时期进行取样，以氨基酸肉汤培养基作为选择培养基对产胺菌株进行筛选，利用高效液相色谱对其产胺能力进行定量分析，挑选出高产胺菌株，探究其在不同环境(pH、盐度、温度)下的产胺能力。共筛得26株产胺菌株，经16S rRNA基因测序技术鉴定为8个属，包括肠杆菌属、克雷伯氏菌属、勒克氏菌属、假柠檬酸盐杆菌属、莱略特菌属、泛菌属、魏斯氏菌属和乳杆菌属，其中肠杆菌科菌株具有很强的产胺能力，总产胺量最高为7 400.13 mg/L,主要产胺种类为腐胺和尸胺。温度对于菌株产胺能力的影响相对较小，而pH和盐度对菌株产胺能力有明显的影响，当pH值低至3.5、盐度高达80 g/L时，菌株的产胺能力受到强烈抑制。研究结果可为控制泡菜中产胺微生物生长，降低生物胺积累提供理论参考。     

一种混合发酵剂对干发酵香肠中生物胺含量的影响

研究了复合发酵剂(戊糖片球菌Pediococcuspentocaceus和木糖葡萄球菌Staphylococcusxylosus)对发酵香肠中生物胺含量的影响。通过与单一发酵剂戊糖片球菌和空白组的产胺、微生物生长情况的比较,分析了木糖葡萄球菌对戊糖片球菌在发酵过程中降胺作用的影响。结果表明,木糖葡萄球菌有助于戊糖片球菌进一步降低发酵香肠中的酪胺和色胺的量,分别比单一发酵菌株降低了6.7%、22.83%;三种胺的量比空白组分别降低了94.04%,29.53%,46.59%,从而证实了戊糖片球菌具有降酪胺和色胺的能力。结合微生物菌数可知,木糖葡萄球菌是通过抑制乳酸杂菌的生长促使戊糖片球菌菌数的增加而有利于胺量的降低。     

西式发酵香肠中生物胺含量的调查分析

综述生物胺的种类、危害及在西式发酵香肠中存在的可能,采集了西式发酵香肠的样本,检测了其5种生物胺(尸胺、组胺、酪胺、亚精胺、精胺)及4种微生物(乳酸菌、微球菌、肠细菌、假单胞菌)的含量。经分析得出结论:5种生物胺在西式发酵香肠中普遍存在;肠细菌的数量与生物胺的含量有显著的相关性,未发现其他3种微生物与生物胺含量之间的关系。     

不同发酵剂对发酵香肠中风味物质释放及有害生物胺控制的影响

通过接种植物乳杆菌与肉葡萄球菌,将试验分为复合发酵组和单一植物乳杆菌组,并以自然发酵为对照。探究发酵剂对发酵香肠加工过程中pH、Aw、色泽、亚硝酸盐、风味物质释放及有害生物胺含量控制的影响。结果表明:复配组产酸速率单一组产酸速率对照组产酸速率,且干燥结束时复配组降到组中最低为4.74;pH快速降低也使复配组Aw下降速率显著快于其它两组(P0.05);9 d后成熟结束时,复配组亚硝酸盐含量为12.6 mg/kg,显著低于其它两组及国家安全规定30 mg/kg(P0.05);从3组香肠中检出尸胺、β-苯乙胺、酪胺、组胺及精胺和亚精胺,由于精胺和亚精胺是肉中固有生物胺且含量变化微小,所以在试验中未加以说明;成品时所列4种生物胺总量在33.37～40.41 mg/kg范围,复配组中组胺及酪胺含量分别为2.89,22.54 mg/kg,低于美国食品及药品管理局(FDA)规定标准(组胺含量50 mg/kg,酪胺含量≤100 mg/kg)(P0.01)。从香肠中共检出26种挥发性风味物质,其中添加微生物发酵组风味物质总含量显著高于对照组(P0.01)。综上,复配发酵剂可使pH、Aw快速降低,有效抑制和降低香肠残留亚硝酸盐及有害生物胺含量,对香肠中风味物质释放和含量增加起主要作用。     

Contribution of contaminant enterobacteria and lactic acid bacteria to biogenic amine accumulation in spontaneous fermentation of pork sausages.

The contribution of selected contaminant bacteria to the qualitative and quantitative profile of biogenic amine formed was examined during the spontaneous fermentation of pork sausages (without starter culture). The study mainly dealt with the evolution of the amine profile and microbial development during spoilage of meat, but also focused on the biogenic amine build-up during sausage fermentation using pork meat contaminated by a tyramine-positive Lactobacillus brevis strain and a cadaverine-positive Enterobacter cloacae strain. Amines and microbial counts correlated well during meat spoilage and sausage fermentation. The high accumulation of cadaverine was associated with the undesirably high number of Enterobacteriaceae. Tyramine production during sausage fermentation was not entirely attributable to contaminant lactic acid bacteria. On the contrary, the spontaneous fermenting flora needed to produce these types of product are also involved. The contribution of contaminant bacteria to the putrescine and other biogenic amines during meat fermentation remains unclear.     

不同发酵方式对发酵马肉肠贮藏期品质劣变及食用安全性的影响

为有效确保发酵马肉肠贮藏期间的品质及安全性,研究添加汉逊德巴利酵母菌1808（Debaryomyces hansenii 1808）、木糖葡萄球菌21445（Staphylococcus xylose 21445）、植物乳杆菌E11（Lactobacillus plantarum E11）的复合发酵剂发酵组（FP）和单一乳酸菌发酵组（DZ）、自然发酵组（ZR）马肉肠贮藏过程中生物胺的变化,考察不同发酵方式对发酵马肉肠pH值、水分活度（aw）、色差、总挥发性盐基氮（total volatile basic nitrogen,TVB-N）含量、硫代巴比妥酸反应物（thiobarbituric acid reactive substances,TBARs）值及菌落总数的影响。结果表明：在贮藏期间,相比于自然发酵组和单一乳酸菌发酵组,添加复配发酵剂可有效减缓发酵马肉肠TVB-N含量、TBARs值、pH值的增幅,抑制贮藏后期有害杂菌的生长,防止产品过早发生腐败变质;此外,复配发酵剂可显著降低贮藏过程中马肉肠的水分损失（P＜0.05）,维持色泽的稳定,抑制生物胺（尸胺、腐胺、酪胺、组胺）的积累。综上,随着贮藏时间的延长,复配发酵剂可显著抑制发酵马肉肠品质的劣变及不良生物胺的过多积累。     

Species diversity and metabolic impact of the microbiota are low in spontaneously acidified Belgian sausages with an added starter culture of Staphylococcus carnosus

Quality of fermented sausages is affected by acidifying lactic acid bacteria (LAB) and colour- and flavour-promoting coagulase-negative staphylococci (CNS), whether or not used as starter culture. Artisan fermented sausages are often perceived as superior to industrial variants, partially because of the specific microbiota due to spontaneous acidification, which may be considered as an artisan characteristic. Therefore, two kinds of spontaneously acidified Belgian sausages were prepared (Belgian-type salami and Boulogne sausage), but with addition of a Staphylococcus carnosus culture. The Belgian-type salami was made from pork and beef, whereas the Boulogne sausage contained pork and horse meat. In all cases, Lactobacillus sakei was the dominant LAB species present on the raw materials and during fermentation, whereas enterococci remained present in the background. Enterobacteriaceae vanished after fermentation. The CNS species diversity on the raw materials was large and differed between the pork, beef, and horse meat. Nevertheless, this species diversity was annihilated during fermentation by the added S. carnosus culture. The volatiles fraction was mainly composed of aldehydes that originated from lipid oxidation and spices-derived compounds. Aromatic compounds that are typically associated to CNS activity, such as end-products from the metabolism of branched-chain amino acids, were not present in the Belgian-type salami and only marginally present in the Boulogne sausage. In conclusion, spontaneous acidification of Belgian-type fermented sausages leads to dominance of L. sakei and is no guarantee for bacterial contribution to the aroma profile when S. carnosus is added as a starter culture.     

Effectiveness of a Lactobacillus sakei starter culture in the reduction of biogenic amine accumulation as a function of the raw material quality

The effectiveness of an amine-negative starter culture (Lactobacillus sakei CTC494) in the reduction of biogenic amine production during the ripening of fermented sausages was examined. Four batches were manufactured in parallel: spontaneously fermented and starter-mediated sausages were manufactured from two lots of raw materials of different hygienic quality. Besides the biogenic amine contents, changes in the microbial counts, nitrogenous fractions, pH, and water content were measured at several sampling points during the ripening process. In sausages manufactured from good quality meat, the starter strain of L. sakei reduced and even inhibited biogenic amine accumulation during sausage fermentation, the end products showing extremely low biogenic amine contents (tyramine levels less than 15 mg/kg of dry matter and putrescine and cadaverine levels less than 5 mg/kg of dry matter). Nevertheless, starter-mediated sausages made from poorer-quality raw materials showed much higher amine contents (308, 223, and 36 mg/kg of dry matter of cadaverine, tyramine, and putrescine, respectively), which were only slightly lower than those of the spontaneously fermented sausages made from the same raw materials. The relatively high bacterial numbers of raw materials of poorer-hygienic quality diminished the beneficial effect of the starter strain. Therefore, the effectiveness of the starter was strongly dependent on the hygienic quality of the raw materials used.     

Reduction of biogenic amine formation using a negative amino acid-decarboxylase starter culture for fermentation of Fuet sausages

The ability of Lactobacillus sakei CTC494, a negative amino acid-decarboxylase starter culture, to reduce biogenic amine accumulation during sausage fermentation and storage at 4 and 19 degrees C was studied. The effect on the amine formation of the tyramine producer Lactobacillus curvatus CTC371, as a positive strain, was also examined in comparison to a spontaneous fermentation process without starter culture (control batch). The polyamines spermine, spermidine, and diaminopropane were not influenced by the ripening, and their levels slightly decreased in all the batches throughout the storage. Tyramine, cadaverine, and putrescine were the main amines formed during the ripening. The addition of starter culture resulted in a decrease on the biogenic amine formation, depending on the strain inoculated. A great reduction in tyramine content was achieved when L. sakei CTC494 was inoculated, whereas L. curvatus CTC371 only attenuated tyramine accumulation compared with the control batch. Both starters were able to significantly limit the production of putrescine and cadaverine, and they inhibited tryptamine and phenylethylamine formation by the wild microbial flora. Tyramine levels of the control sausages rose during the storage at both temperatures, whereas those of cadaverine only increased at 19 degrees C. On the contrary, sausages manufactured through the starter controlled fermentation did not show changes of amine contents during the storage. The addition of a proper selected starter culture is advisable to produce safer sausages with low contents of biogenic amines.     

冷却猪肉及托盘表面细菌生物被膜分析和肉源荧光假单胞菌的鉴定与被膜研究

为研究冷却猪肉及其接触面中细菌生物被膜能力并探讨肉类特定腐败菌的生物被膜特征,分析了腐败的冷却肉和销售托盘表面的细菌生物被膜形成能力;利用形态学观察、16S r DNA分析和VITEK2微生物鉴定系统鉴定了冷却肉中一株生物被膜能力较强的荧光假单胞菌(Pseudomonas fluorescens);测定了该菌在培养基和猪肉浸提液中的生物被膜能力;用激光共聚焦和扫描电子显微镜观测了其生物被膜的结构特征。结果发现:冷却肉中及其销售托盘表面的细菌能够形成生物被膜的比例较高,37%和45%的细菌具有较强生物被膜形成能力。肉源性荧光假单胞菌能够在培养基和肉液中形成大量生物被膜,具有较强的生物被膜能力。显微成像结果表明荧光假单胞菌在培养6 h后有明显黏附,18 h后多糖分泌增多、菌体堆积并形成具有生物功能的立体生物被膜。这些特点可能有利于荧光假单胞菌在冷却肉表面黏附并成为优势腐败菌。     

Biogenic amine formation in Nham,a Thai fermented sausage,and the reduction by commercial starter culture, Lactobacillus plantarum BCC 9546

Biogenic amines are of concern for sausage due to their toxicological effects on nervous, blood pressure, gastric and intestinal systems. In this study, the influence of raw pork meat quality and starter culture inoculation on biogenic amines accumulation in Nham, a Thai traditional fermented pork, were studied. Before Nham processing, pork meat was stored at 30 °C for 6 h, and at 4 and −20 °C for 2 days. Formation of biogenic amines (cadaverine, putrescine, histamine and tyramine) was significantly higher in Nham processed from stored meat. Accumulation of these biogenic amines in Nham reduced significantly by the addition of Lactobacillus plantarum BCC 9546, a commercial Nham starter culture. The results highlight the importance of using fresh meat products and the inclusion of an appropriate starter culture to minimise the formation of biogenic amines during the process of Nham fermentation.     

不同储藏时期香肠菌群结构的变化及优势腐败菌研究

以天然肠衣的熏煮香肠为研究对象,将新鲜香肠和腐败香肠分别取样,提取DNA,采用高通量测序技术对两种香肠的菌群结构进行比较研究,并确定优势腐败菌。结果表明:腐败香肠菌群丰度和均匀度较新鲜香肠降低;在属的水平上,新鲜香肠中丰度最高的是Staphylococcus(葡萄球菌属90.64%),而腐败香肠中丰度最高的是Weissella(魏斯氏菌属65.93%),其次是Staphylocococcus(葡萄球菌属33.79%);在种的分类水平上,新鲜香肠中Staphylococcus equorum(马胃葡萄球菌25.59%)和Staphylococcus vitulinus(小牛葡萄球菌18.01%)丰度最高,腐败香肠中丰度最高的是Weissella viridesecens(绿色魏斯氏菌46.04%),其次是Weissella cibaria(食窦魏斯氏菌19.68%)。结果提示:与新鲜香肠的菌群组成相比较,腐败香肠的菌群结构发生了显著的变化;腐败香肠中的优势腐败菌是Weissella viridesecens,该种微生物在香肠储藏过程中大量增殖,主导了产品的腐败;在生产过程中需消除Weissella viridesecens,以延长产品货架期,提高产品品质。     

发酵肉制品中生物胺及其产生菌株研究进展

生物胺是一类具有生物活性的含氨基小分子有机化合物,其广泛存在于蛋白丰富的食品中。生物体内适量生物胺具有增强代谢、促进生长等功能,但当含量达到临界值时,会产生诸多不良影响。存在大量微生物的食品中,生物胺的含量较高,同类发酵肉制品中生物胺的种类和含量也有很大不同,如发酵时间、贮藏时间和包装材料等都会对此产生影响。本文主要介绍生物胺的常见种类、生理作用,发酵肉制品中产生物胺的微生物种类、形成途径、菌株筛选方法及检测方法,旨在为今后发酵肉制品中生物胺的进一步研究提供帮助,以期为发酵肉制品的质量保障提供参考。     

Evaluating the Influence of Malolactic Fermentation Inoculation Practices and Ageing on Lees on Biogenic Amine Production in Wine

Biogenic amines are compounds, produced primarily by lactic acid bacteria (LAB) that negatively affect the wholesomeness of wine. Standard winemaking practices can greatly influence the levels of biogenic amines in wine. The aim of this study was to determine the relative contribution of different malolactic fermentation (MLF) practices and ageing of wines on fermentation lees to the final levels of biogenic amines. Wines were made on small scale over two harvest seasons with two red grape cultivars. Treatments included spontaneous MLF, co-inoculated MLF, MLF inoculated after alcoholic fermentation (conventional inoculation) and 4 months of ageing in the presence and absence of fermentation lees of all MLF treatments. Biogenic amine concentrations were measured by high-performance liquid chromatography at key winemaking stages and statistically analysed for the effects of MLF treatment and winemaking stage. Results indicate that the presence of indigenous LAB increased the risk of biogenic amine formation. Inoculation proved to reduce biogenic amine production over time compared to spontaneous MLF and co-inoculation even more than conventional inoculation. The presence of yeast lees during ageing generally led to higher final concentrations of biogenic amines in wines than the absence of lees. This study confirms other works that conclude that spontaneous MLF and uncontrolled ageing on yeast lees are generally unpredictable and pose a risk of biogenic amine contamination in finished wines.