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。     

Biogenic amine and microbiological profile of Serbian dry fermented sausages of artisanal and industrial origin and associated health risk

ABSTRACT

From the public health point of view, safety of dry fermented sausages is of concern due to possible presence of biogenic amines and some other food hazards. Taking into consideration that biogenic amines are chemical indicators of microbiological contamination, industrial and artisanal dry fermented sausages produced in Serbia were subjected to the microbiological and biogenic amine profiling. Microorganisms identified by MALDI-TOF MS included Cronobacter sakazakii and Listeria innocua, whereas the food pathogens were not detected. Biogenic amine content, determined by HPLC method in the range from 37.3 to 1186 mg/kg, was characterised as very low to low in 40% of the samples, moderate in 38%, high in 14% and very high in 8%. Risk assessment revealed that consumption of dry fermented sausages could cause histamine and tyramine intake up to 11.9% and 3.4% of threshold dose for healthy population, respectively. Adverse health effects would be rather unlikely for general population.     

Relationship between biogenic amine contents and the size of dry fermented sausages

Three trials were carried out to study the influence of the diameter on biogenic amine contents and related parameters (pH, humidity and proteolysis) in fermented sausages. The first trial was done on three groups of Spanish dry fermented sausages with different diameter. In the second, two sections (centre and edge) of salchichón sausages were examined. The last trial consisted in the study of the ripening of two batches of sausages fermented under the same conditions but with two different diameters. Biogenic amine contents varied among the different type of products as well as among the same type of samples. Generally, amine levels in the biggest diameter sausages were higher than in the thinnest sausages. Amine contents were higher in the central part of the sausages than in the edge. During the ripening, larger tyramine amounts were formed in sausages with the biggest diameter. Statistical correlations were found among the diameter, the pH, the proteolysis and some amines. The results of the three trials agree with the hypothesis that the diameter is a factor that may affect the formation of biogenic amines during sausage fermentation.     

Biogenic amines in traditional fermented sausages produced in selected European countries

Aminogenesis in traditional fermented sausages produced in Europe was studied during manufacturing process taking into account technological, physico-chemical and microbial factors. Tyramine was the major amine, followed by putrescine and cadaverine, although the occurrence of di-amines was much more variable. By principal component analysis, relationships between aminogenesis and the country of origin, physico-chemical parameters, processing conditions and microbial counts, were not found, probably due to the high dispersion observed in those variables. Therefore, biogenic amines occurred irrespectively of physico-chemical changes and technological conditions applied for sausage manufacture. By cluster analysis, five groups of fermented sausages were identified on the basis of their quantitative and qualitative profile of total biogenic amine content. Group A included products from very low to low total amine content (from not detected to 150 mg/kg); group B, products with moderate levels (from 150 to 350 mg/kg) tyramine being the major amine; group C, also with moderate amine contents but cadaverine being the major amine; and groups D and E, comprising products with high (from 350 to 550 mg/kg) and very high (higher than 550 mg/kg) amine content, respectively. Samples with moderate, high or very high levels of biogenic amines could be considered as products of less quality, and their consumption could be unhealthy for sensitive individuals or for those under classical monoamine oxidase inhibitor drug therapy.     

发酵香肠中生物胺的研究进展

生物胺是一种低分子量的有机物,主要是通过氨基酸的脱羧作用生成.它广泛存在于发酵香肠、泡菜、干酪、酸奶等发酵食品中.本文综述了目前国内外关于发酵香肠中生物胺的研究情况,主要介绍了生物胺的产生原因、常见的生物胺种类、产生物胺的主要微生物、影响生物胺产生的理化因素以及对发酵香肠中生物胺含量积累的控制措施,旨在为今后对发酵香肠中生物胺的进一步研究提供帮助.     

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.     

Biogenic amine content of boza: A traditional cereal-based, fermented Turkish beverage

Boza is a fermented beverage made from millet, maize, wheat or rice. Biogenic amine contents of 10 boza samples from different manufacturers in Turkey were analysed for the first time, using HPLC after derivatisation with benzoyl chloride. Of the 11 biogenic amines under study, putrescine, spermidine and tyramine were detected in all boza samples. Tyramine was the prevailing biogenic amine. Tyramine concentrations of boza samples were between 13 and 65 mg/kg. Total biogenic amine contents of boza samples were between 25 and 69 mg/kg. Consequently, consumption of boza might represent a health risk for patients being treated with drugs containing monoamine oxidase inhibitor (MAOI). The pH values of boza samples were in the range from 3.16 to 4.02; total dry matters were from 15.3% to 31.1% (w/w); protein contents were from 0.50% to 0.99% (w/w). No significant correlations were detected between biogenic amine concentrations and pH, protein content and total dry matter content.     

Determination of Biogenic Amines in Sichuan-Style Spontaneously Fermented Sausages

The qualitative and quantitative profile of biogenic amines in 30 samples of Sichuan-style spontaneously fermented sausages sold in Sichuan retail markets was studied by HPLC. Tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, and spermidine were analyzed by UV detection after pre-column derivatization with dansyl chloride. There were wide ranges of different biogenic amines in the samples, except spermidine. The results showed that the histamine was the major biogenic amines, followed by tyramine and cadaverine, which have a mean value of 196.06 mg/kg (ranging from 114.33 to 327.35 mg/kg), 164.67 mg/kg (ranging from 88.84 to 285.89 mg/kg), and 141.65 mg/kg (ranging from 19.14 to 376.46 mg/kg), respectively. The toxicity limit of histamine is 100 mg/kg, where all of the samples exceeded this limit. None of the samples of total biogenic amines content exceeded 1000 mg/kg. The results showed that the amounts of biogenic amines in Sichuan-style spontaneously fermented sausages sold in Sichuan retail markets may pose a potential health risk for sensitive individuals. This suggested that it is necessary to improve the production techniques and the raw materials’ hygienic quality, to ensure the quality and safety of this fermented sausages with respect to their biogenic amine content.     

原料肉贮藏时间对牛肉发酵香肠中微生物菌相变化和蛋白质降解的影响

采用传统微生物计数、高通量测序、凯氏定氮、高效液相色谱等方法,探究牛肉宰后0～4 ℃下不同贮藏时间（0、12、24、36 h）对发酵香肠加工过程中微生物菌相变化、蛋白质降解及其相关性的影响。结果显示,随着牛肉贮藏时间的延长,香肠中菌落总数、大肠菌群数、菌群多样性和丰富度以及蛋白质降解指标均呈上升趋势,总游离氨基酸含量呈下降趋势;香肠成熟阶段中总挥发性盐基氮含量与总生物胺含量之间呈极显著正相关（P＜0.01）。其中,贮藏36 h牛肉加工的香肠与未经贮藏牛肉加工的香肠对比,菌落总数平均值由2.40×107 CFU/g增加到4.03×107 CFU/g（P＜0.05）,反映菌群多样性的Shannon指数平均值由1.39增长至2.89,优势菌群由整个加工阶段均为葡萄球菌变成前期为魏斯氏菌,后期为葡萄球菌;总游离氨基酸含量平均值由7.09 mg/g减少至6.44 mg/g,总生物胺含量平均值由92.14 mg/kg增长为117.42 mg/kg（P＜0.05）。结论：控制原料牛肉贮藏时间可以有效减少香肠中微生物的总量,降低菌群多样性,抑制蛋白质的降解,减缓生物胺的积累,从而保障发酵香肠的安全性。     

发酵香肠中生物胺控制技术的研究进展

生物胺是一类具有生物活性的低分子量含氮有机化合物的总称,适量生物胺有助于人体正常的生理功能,但生物胺含量过高不仅会降低发酵香肠品质,而且会对机体健康造成不良反应。本文综述了发酵香肠中生物胺的种类和危害性并重点介绍了生物胺的物理、化学和生物控制方法,展望了发酵香肠中生物胺的控制技术的研究方向。      

Biogenic amine contents in selected Egyptian fermented foods as determined by ion-exchange chromatography

Biogenic amines in foods may pose a public health problem. Therefore, histamine, tyramine, putrescine, cadaverine, spermine, and spermidine concentrations were measured in selected food items commonly consumed in Egypt. Foods examined were dairy products (blue cheese and Mesh cheese), meats (fermented and smoked cooked sausage), and fish (smoked and salted fermented fish [Feseekh], salted sardines, and anchovies). Egyptian fermented sausage had the highest concentration of total biogenic amines (2,482 mg/kg), followed by Mesh cheese (2,118 mg/kg) and blue cheese (2,084 mg/kg). The lowest concentration was found in smoked cooked sausage (111 mg/kg). Histamine was found at a high level (521 mg/kg) in Feseekh, and tyramine was highest (2,010 mg/kg) in blue cheese. These results indicate that some traditional Egyptian foods may pose a health risk due to the concentration of biogenic amines, especially histamine.     

Influence of hygienic quality of raw materials on biogenic amine production during ripening and storage of dry fermented sausages

The effect of the hygienic status of raw materials on biogenic amine production during ripening and storage (at 4 and 15 degrees C) of fermented sausages was studied. Two portions of fresh lean and back fat pork were stored for 5 days at -20 degrees C (treatment A) and at 4 degrees C (treatment B), respectively. Raw materials of treatment A maintained their hygienic quality high and low amine content. Raw materials of treatment B showed from 1 to 3 log (CFU/g) higher microbial counts and a biogenic amine index near 50 mg/kg, indicating poorer hygienic quality. The quality of raw materials influenced the composition and the concentration of biogenic amines produced during the ripening sausages. Sausages of treatment A (A-sausages) showed a large accumulation of tyramine (up to 100 mg/kg dm) followed by putrescine and cadaverine (<15 mg/kg). In contrast, B-sausages resulted in earlier and much greater amine production, and cadaverine, tyramine, and putrescine levels were 50-, 2.6-, and 6.5-fold higher than those of A-sausages. Other biogenic amines, such as octopamine, tryptamine, phenylethylamine, and histamine were also produced in B-sausages. The higher proteolysis and the lower pH of B-sausages might have favored the decarboxylase activity of microorganisms. Biogenic amine contents of sausages during storage depended on the raw materials used and storage temperature. No significant modification on the amine contents was observed during the storage of A-sausages at either temperature. Greater changes occurred in B-sausages stored at 15 degrees C than in those stored at 4 degrees C. Higher temperatures favored proteolytic and decarboxylase reactions, resulting in increased amine concentrations after storage.     

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.     

复合果蔬取代部分硝酸盐对降低发酵香肠中亚硝胺的作用

分析复合果蔬替代硝酸盐对发酵香肠中N-亚硝胺生成的影响,探究发酵香肠生产过程中复合果蔬取代部分硝酸盐的可能性。制备4 组中式发酵香肠：阴性对照组（negative control group,NCG,空白组）、阳性对照组（positive control group,PCG）、芹菜樱桃组（celery and cherry group,CCG）、卷心菜番茄组（cabbage and tomato group,CTG）,考察在发酵及成熟过程中亚硝酸盐、亚硝胺及生物胺含量的变化。结果表明：发酵过程中,果蔬发酵香肠（CCG和CTG）与PCG亚硝酸盐含量同样呈现先增加后降低趋势,但CTG峰值（65.52?mg/kg）显著低于PCG（105.31?mg/kg）。4?组发酵香肠均能检出N-二乙基亚硝胺（N-nitrosodiethylamine,NDEA）,而果蔬发酵香肠（CCG和CTG）在发酵后期NDEA含量（1.23 μg/kg未检出）显著低于PCG（3.72 μg/kg）。干燥成熟阶段,果蔬发酵香肠（CTG和CCG）亚硝酸盐残留量（36.45?mg/kg和47.97?mg/kg）显著高于NCG（24.49?mg/kg）和PCG（31.40 mg/kg）,但在干燥3 d后下降至国家标准限量（30 mg/kg）以下。成品中总生物胺含量排序为NCG＞PCG＞CCG＞CTG,CTG中未检出组胺和酪胺。各组样品中N-二甲基亚硝胺（N-nitrosodimethylamine,NDMA）（PCG除外）和NDEA含量均随干燥进程而逐渐增加,但是果蔬发酵香肠（CCG和CTG）在干燥结束时NDMA（0.16?μg/kg和0.11?μg/kg）和NDEA含量（4.33?μg/kg和4.13?μg/kg）低于NCG（1.73?μg/kg和9.50?μg/kg）,与PCG（0?μg/kg和4.74?μg/kg）相当。实验表明复合果蔬浆替代部分硝酸盐,可降低中式发酵香肠的N-亚硝胺和生物胺含量。     

不同发酵豆制品中生物胺调查分析

目的 调查腐乳、豆酱和豆豉3类豆制品中生物胺含量, 初步了解发酵豆制品中生物胺存在情况。方法 参照GB 5009.208-2016《食品安全国家标准 食品中生物胺的测定》, 采用高效液相色谱法测定6种腐乳、3种豆酱、6种豆豉中的生物胺含量。结果 腐乳中的生物胺含量明显超出另两类, 其总生物胺平均含量达472.35 mg/kg; 豆酱略高于豆豉, 分别为196.77 mg/kg和171.46 mg/kg。腐乳、豆豉和豆酱样品中含量最高的生物胺均是酪胺, 腐乳样品中居其次的是腐胺, 豆豉和豆酱样品中是章鱼胺, 3种品种样品亚精胺和精胺含量均较少。结论 从整体而言, 3类发酵豆制品生物胺含量在安全范围内。     

Use of Fourier transform infrared spectroscopy to evaluate the proteolytic activity of Yarrowia lipolytica and its contribution to cheese ripening

The interaction between tyrosine-decarboxylase and proteolytic activities of a Lactobacillus curvatus and Staphylococcus xylosus, respectively, on biogenic amine production during the ripening and the storage of dry fermented sausages was investigated. Water content, pH, proteolysis parameters, microbial counts, and biogenic amine contents were monitored in spontaneously and starter fermented sausages. The use of proteolytic staphylococci as starter resulted in a higher content of non-protein nitrogen and total free amino acids. Tyramine was the main amine produced in all batches. However, tyrosine-decarboxylase activity of the L. curvatus starter strain was weak and yielded lower amounts of tyramine than those produced by the wild mioroflora in the control batch. Association between tyramine production and proteolysis could only be established in a defectively dried batch. Putrescine and cadaverine accumulation was efficiently reduced in the starter-mediated fermentation, in agreement with the lower development of enterobacteria. Phenylethylamine and tryptamine were only detected in the spontaneously fermented sausages, while histamine, spermine and spermidine did not vary during the ripening. Biogenic amine levels and related parameters showed significant changes during the storage of dry sausages depending on the temperature and the batch. As a general rule, changes in the pH, proteolysis, microbial counts, and biogenic amine contents were stronger at 19 degrees C than at 4 degrees C. The results suggest that refrigeration would be advisable for preventing further accumulation of biogenic amines during the storage of dry fermented sausages.     

Changes in biogenic amine and polyamine contents in slightly fermented sausages manufactured with and without sugar

The effect of sugar omission on biogenic amine contents of slightly fermented sausages during ripening and storage was evaluated. Two batches of spontaneously fermented sausages were produced with and without sugars in two different trials at pilot-plant scale. Ripened sausages were stored at 4 and 19°C for a further 20 days. Tyramine and cadaverine were the main amines formed during ripening, their contents being significantly higher in batches without sugar. High counts of LAB and Enterobacteriaceae could be associated with the production of tyramine and cadaverine, respectively. The occurrence of putrescine depended on the trial and batch. Sausages without sugar contained more putrescine than those with sugar in trial 1, but this was not repeated in trial 2, in which a high production of agmatine occurred. Tryptamine and phenylethylamine were only detected in the later stages of ripening, their contents also being higher in sausages without sugar. Biogenic amine contents generally rose during storage at 19°C, the increase being especially important for cadaverine and tyramine in sausages without sugar. Levels of spermidine and spermine remained constant during ripening and decreased slightly during storage. Sugar omission is not recommended because it might increase biogenic amine accumulation during the manufacture and storage of slightly fermented sausages.     

Changes in biogenic amine levels during storage of Mexican-style soft and Spanish-style dry-ripened sausages with different aw values under modified atmosphere

Two raw sausages were prepared: a soft and a dry-ripened one, both by local traditional and industrial manufacturing practices. Sausages were packaged under a CO₂/N₂ atmosphere at different targeted activity water (a_w) values: 0.96 and 0.92 (soft sausages) and 0.88 and 0.82 (dry-ripened sausages). Sausages were then stored at 5 °C for 42 days or at 12 °C for 240 days (soft and a dry-ripened sausages, respectively). The time-related changes in dominant microbiota, pH and biogenic amine contents during storage were determined. Tyramine was the most abundant biogenic amine in all the sausages. Biogenic amine levels were higher in dry-ripened sausages than in soft sausages at packaging. However, during refrigerated storage soft sausages were fermented and the levels of biogenic amines increased (P < 0.05). At the end of storage, traditional soft sausages with 0.96 a_w presented comparable levels of biogenic amines to traditional dry-ripened sausages.     

Biogenic amines in dry fermented sausages: a review

Biogenic amines are compounds commonly present in living organisms in which they are responsible for many essential functions. They can be naturally present in many foods such as fruits and vegetables, meat, fish, chocolate and milk, but they can also be produced in high amounts by microorganisms through the activity of amino acid decarboxylases. Excessive consumption of these amines can be of health concern because their not equilibrate assumption in human organism can generate different degrees of diseases determined by their action on nervous, gastric and intestinal systems and blood pressure. High microbial counts, which characterise fermented foods, often unavoidably lead to considerable accumulation of biogenic amines, especially tyramine, 2-phenylethylamine, tryptamine, cadaverine, putrescine and histamine. However, great fluctuations of amine content are reported in the same type of product. These differences depend on many variables: the quali-quantitative composition of microbial microflora, the chemico-physical variables, the hygienic procedure adopted during production, and the availability of precursors. Dry fermented sausages are worldwide diffused fermented meat products that can be a source of biogenic amines. Even in the absence of specific rules and regulations regarding the presence of these compounds in sausages and other fermented products, an increasing attention is given to biogenic amines, especially in relation to the higher number of consumers with enhanced sensitivity to biogenic amines determined by the inhibition of the action of amino oxidases, the enzymes involved in the detoxification of these substances. The aim of this paper is to give an overview on the presence of these compounds in dry fermented sausages and to discuss the most important factors influencing their accumulation. These include process and implicit factors as well as the role of starter and nonstarter microflora growing in the different steps of sausage production. Moreover, the role of microorganisms with amino oxidase activity as starter cultures to control or reduce the accumulation of biogenic amines during ripening and storage of sausages is discussed.