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。     

发酵香肠中生物胺生成量的菌株效应分析

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)在发酵香肠中生物胺的生成量进行了分析。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。     

Effect of starter culture, spice mix and storage time and temperature on biogenic amine content of dry fermented sausages

Two types of dry fermented sausage differing in spicing mixture and the diameter (low content of red pepper + diameter 80 mm, H-sausage; high content of red pepper + diameter 55 mm, P-sausage, respectively) were produced in parallel with two different starter cultures (Pediococcus pentosaceus + Staphylococcus carnosus, B-samples and S. carnosus + Staphylococcus xylosus + Lactobacillus farciminis, F-samples, respectively). The sausages were ripened 21 days and subsequently stored 91 days at the room temperature. Concentration of both most abundant amines, putrescine and tyramine (y; mg/kg DM) increased significantly (P<0.01) in HB-sausage during ripening (x; days): y=2.5+18.13x−0.3144x² (R²=0.99) and y=0.7+8.17x−0.1130x² (R²=0.99), and also during storage: y=127.3+5.123x (R²=0.79) and y=26.0+3.211x (R²=0.74), respectively. At the end of ripening, putrescine (247 mg/kg DM) and tyramine (123 mg/kg DM) content in the HB-sausage was higher (P<0.05) than in the PB-sausage (12 and 9 mg/kg DM, respectively), concentration of either of these amines was negligible (1 mg/kg DM) in either type of F-inoculated sausage. Both starter culture and sausage type influenced significantly (P<0.001) both putrescine and tyramine content in the sausage; starter accounted for 57% and 55% of total variability in putrescine and tyramine content, respectively. Due to the significant (P<0.05) increase of total aerobic counts in the HB-sausage between the end of ripening and the 7th day of storage, followed by the significant (P<0.01) increase of the sum of total biogenic amines between the end of ripening (425 mg/kg DM) and the end of storage (1029 mg/kg DM), the storage of the dry fermented sausages at the room temperature should not be recommended.     

FORMATIONS OF BIOGENIC AMINES IN TURKISH STYLE SAUSAGE DURING RIPENING AND STORAGE PERIODS

Changes of biogenic amine (histamine, β‐phenylethylamine, serotonin, spermine, spermidine, cadaverine, putresine, 1,7‐diamino heptane, tryptamine and tyramine) concentrations and pH values of Turkish style sausages were investigated during ripening and storage. Cadaverine and β‐phenylethylamine were not detected during the ripening and storage periods for all sausage types. Storage time had a significant (P<0.05) effect on the formation of all biogenic amines. Starter culture usage decreased (P<0.05) biogenic amine formation. Histamine levels were higher in sausages prepared without additives and lower in the sausages made with starter culture than the sausages made without starter culture. There was no effect of starter culture and additives on formation of tryptamine, 1,7‐diamino heptane, serotonin or spermidine. Starter culture and additives affected (P<0.05) the formation histamine, putrescine and spermine. pH values of sausages sharply decreased (P< 0.05) from 5.98 to about 4.53 during 3 days of ripening. After that, pH values increased slowly and were nearly constant during the storage period.     

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.     

Effects of starter cultures and nitrite levels on formation of biogenic amines in sucuk

The effects of starter cultures, starter A (Lactobacillus sakei+Staphylococcus carnosus) and starter B (Pediococcus acidilactici+Staphylococcus xylosus+Lactobacillus curvatus), nitrite levels (0, 75, and 150ppm) and ripening period on biogenic amine production were investigated in sucuk (Turkish dry-fermented sausage) production. Changes in biogenic amines, pH, water activity, moisture, non-protein nitrogen, nitrite and counts of lactic acid bacteria, Micrococcus/Staphylococcus, Enterobacteriaceae, and moulds-yeasts were determined. Ripening period had a very significant effect (P<0.01) on the all biogenic amines. Sausages produced by fermentation with starter cultures, as compared to natural fermentation (control), had lower amounts of putrescine, cadaverine and tyramine (P<0.05). Starter cultures A and B were found to have almost the same effect on the formation of biogenic amines. Use of nitrite in sucuk production affected the formation of biogenic amines except for spermidine and spermine (P<0.01). Lactic acid bacteria counts in the control group were lower than that of starter culture groups. Lactic acid bacteria, Micrococcus/Staphylococcus, and moulds-yeasts counts increased during ripening period while levels of species of Enterobacteriaceae decreased, which was also found to be under the detectable level (<100cfug(-1)), by day 3 of ripening.     

Contents of polyamines in selected foods

Dietary polyamines, putrescine, spermidine and spermine, participate in many biochemical processes, mainly in cell proliferation and differentiation. Polyamines were determined as N-benzamides by micellar electrokinetic capillary chromatography in 153 samples of 21 foods, mostly culinary processed. Very low putrescine contents were observed in processed meats, pork liver and kidney, while the highest mean contents exceeded 55 mg kg⁻¹ in stewed green pea, grapefruit and fresh green pepper. Higher spermine than spermidine contents were typical for foods of animal origin, while the opposite was observed in plant products. Mean spermidine contents, exceeding 20 mg kg⁻¹, were found in dry soybean, stewed green pea, yellow pea puree and roasted chicken breast. Roasted chicken breast, stewed pork kidney, roasted pork liver and roasted pork neck had mean spermine contents above the same level. Polyamine content varies widely within individual food items, what makes application of the results by dietitians rather difficult.     

Formation of biogenic amines by Gram-negative bacteria isolated from poultry skin

The aim of this study is to explore production of biogenic amines (histamine, tyramine, putrescine, cadaverine, agmatine, spermine and spermidine) by 88 Gram-negative bacteria isolated from poultry skin (41 isolates of family Enterobacteriaceae, 21 isolates of genus Aeromonas, 16 isolates of genus Pseudomonas, and 10 isolates of other Gram-negative rods). Ion-exchange chromatography was employed to analyse the above mentioned amines. Enterobacteria were found to be the largest producers of amines with proved presence of tyramine, agmatine, putrescine, and cadaverine in cultivation broth after incubation of bacteria. Putrescine and cadaverine were the most abundant products. Presence of at least two biogenic amines, i.e. mainly concurrent presence of putrescine and cadaverine, was revealed in 19 enterobacteria strains. Eleven isolates classified into Aeromonas genus produced putrescine and five of them also formed cadaverine. The other observed biogenic amines (histamine, spermine and spermidine) were not found among tested isolates. Production of biogenic amines by any Pseudomonas family isolates was not proved.     

CHANGES IN BIOGENIC AMINE CONTENTS AND SENSORY QUALITY OF SARDINE (SARDINA PILCHARDUS) STORED AT 4C AND 20C

Biogenic amines (tyramine, putrescine, cadaverine, histamine, spermidine and spermine) in sardine during the storage at two temperatures (4C and 20C) were investigated. Changes in sensory scores were also recorded. Significant differences between storage times were found. The levels of tyramine, putrescine, cadaverine and histamine increased as decomposition of sardine progressed at both 4C and 20C. Spermidine and spermine did not change throughout storage. Putrescine was the main biogenic amine formed, followed by tyramine, cadaverine and histamine. Histamine remained lower than the toxic level of 100 mg/kg in the samples stored at 4C whereas it exceeded the toxic level after 16 h at 20C. The increase in histamine concentration was lower than tyramine and putrescine at both temperatures. Sensory scores gradually decreased during the storage.     

A survey of biogenic amines in chinese red wines

Eight biogenic amines were analyzed in 38 samples of red wine from five wine-making regions of China. The analysis was carried out by reverse phase high performance liquid chromatography with fluorescence detector and dansyl chloride precolumn derivation. Tryptamine was not found in any of the samples. Putrescine was detected in all samples (100%), followed by phenylethylamine (84.2%), spermidine (60.5%), histamine (57.8%), tyramine (57.8%), cadaverine (47.4%), and spermine (36.8%). In all the samples, the levels of aromatic and heterocyclic amines with toxicological effects were: 0–4.58 mg/l for phenylethylamine, 0–10.51 mg/l for histamine, and 0–9.13 mg/l for tyramine. The amount of histamine and tyramine in most of the samples (94.7%) was less than 8 mg/l. The amines associated with sanitary conditions were also found to be present in a very low range, between 0 and 12.98 mg/l for putrescine, 0 and 19.01 mg/l for cadaverine, respectively. In the case of other amines such as spermine and spermidine, they yielded very low levels varying between 0 and 2.64 mg/l for spermine, 0 and 3.82 mg/l for spermidine, respectively.     

熏马肠成熟过程中产生物胺氧化酶菌对生物胺作用的研究

该研究以从新疆熏马肠中筛选出来的6株产生物胺氧化酶菌株(鼠李糖乳杆菌、枯草芽孢杆、腐生葡萄球菌、木糖葡萄球菌、戊糖片球菌、植物乳杆菌)作为发酵剂加入熏马肠中,采用PCR-DGGE技术监测菌落动态变化,高效液相色谱法检测熏马肠中生物胺(色胺、腐胺、苯乙胺、组胺、酪胺、尸胺、精胺、亚精胺)含量变化。结果表明,在熏马肠成熟过程中发酵剂对生物胺有一定的抑制作用。其中以腐生葡萄球菌对色胺、腐生葡萄球菌对苯乙胺、木糖葡萄球菌对尸胺、鼠李糖乳杆菌对腐胺、腐生葡萄球菌对组胺和枯草芽孢杆菌对酪胺的抑制作用最为显著,减少程度分别为45.49%、57.54%、40.75%、51.03%、49.28%、64.33%。本研究为熏马肠以及其他发酵肉制品的提高安全性奠定了基础。     

Relation of biogenic amines with microbial and sensory changes of whole and filleted freshwater rainbow trout (Onchorynchus mykiss) stored on ice

The biogenic amine (BA) content of whole and filleted rainbow trout was monitored during ice storage for a period of 18 days and related to respective microbial and sensorial changes occurring during the same period. Eight amines, namely, putrescine, cadaverine, tyramine, spermidine, tryptamine, beta-phenylethylamine, spermine, and histamine, were determined. Agmatine was not detected in any of the fish samples. In all cases, concentration of BAs was higher (P < 0.05) in filleted compared with whole trout samples. Pseudomonads, H2S-producing bacteria, and, to a lesser extent, Enterobacteriaceae were the dominant microorganisms in both whole and filleted trout. Higher populations (P < 0.05) of these microorganisms were present in filleted trout compared with whole fish samples. Of the BAs determined, concentration of putrescine, cadaverine, spermidine, tryptamine, and beta-phenylethylamine increased steeply in both whole and filleted trout between days 15 and 18 of storage when pseudomonads and H2S-producing bacteria reached approximately 10(6) to 10(7) CFU/g. For the rest of the BAs, including tyramine, histamine, and spermine, a stepwise increase was recorded throughout the entire storage period. Interestingly, Enterobacteriaceae counts remained below 10(6) throughout the entire storage period, accounting for the lower production of histamine. A putrescine value of 13 to 14 mg/kg and a spermidine value of approximately 7 mg/kg for both the whole and filleted trout obtained after 12 and 9 days, respectively, may be proposed as the upper limit for spoilage initiation (freshness indicator) of fresh rainbow trout based on sensorial and microbiological (total viable count of 10(6) to 10(7)) data. With respect to other amines determined, both tyramine and spermine may also be proposed as freshness indicators preferably for whole trout, whereas tryptamine, beta-phenylethylamine, histamine, and cadaverine produced only during later stages of storage are not suitable as freshness indicators of either whole or filleted trout.     

Levels of biogenic amines in typical vegetable products

Biogenic amines were determined in frozen spinach purée, ketchup, concentrated tomato pasta and frozen green pea as N-benzamides, by micellar electrokinetic capillary chromatography. Putrescine and spermidine were observed in most samples at detectable levels, while histamine, spermine and cadaverine concentrations were frequently below detection limits. Maximum mean levels (33.6 and 52.5 mg kg⁻¹ of tyramine and putrescine, respectively) were found in ketchup and 46.6 mg kg⁻¹ of spermidine in pea. Levels of histamine and spermine were low, and those of cadaverine and tryptamine low or medium. Thus, biogenic amines in the observed vegetable products should not be of a risk for healthy consumers.     

发酵剂对牦牛乳硬质干酪成熟过程中生物胺的影响

乳酸菌产生物胺的能力具有菌株特异性,因此,为了探究不同种类发酵剂对牦牛乳硬质干酪中生物胺形成的影响,该试验利用高效液相色谱对3种不同发酵剂制作的硬质干酪成熟过程中生物胺进行了测定和分析。结果表明,嗜热和嗜温发酵剂牦牛乳硬质干酪中检测出2-苯乙胺、腐胺、尸胺、组胺和酪胺,混合发酵剂干酪中检测出腐胺、2-苯乙胺、尸胺和酪胺。各生物胺之间呈现正相关性。3种不同发酵剂干酪在1～6个月成熟过程中,其各生物胺整体呈现增加趋势,嗜热、嗜温和混合发酵剂干酪中总生物胺最高含量分别为(448.3±9.6)、(456.8±58.4)、(293±24.5)mg/kg。组胺和酪胺是2种毒性相对高的生物胺,嗜热发酵剂干酪中组胺和嗜温发酵剂干酪中酪胺最高,其最高含量分别为(20.8±7.9)、(92.9±6.7)mg/kg,混合发酵干酪中未检测出组胺,酪胺含量次之,3种不同发酵剂干酪中组胺、酪胺含量均低于推荐安全剂量50 mg/kg和100 mg/kg。这为合理选择发酵剂和控制干酪中生物胺形成提供了依据。     

Monitoring of biogenic amines in cheeses manufactured at small-scale farms and in fermented dairy products in the Czech Republic

The aim of the study was the monitoring of six biogenic amines (histamine, tyramine, phenylethylamine, tryptamine, putrescine, and cadaverine) and two polyamines (spermidine and spermine) in 112 samples of dairy products purchased in the Czech Republic, namely in 55 cheeses made in small-scale farms and in 57 fermented dairy products. The products were tested at the end of their shelf-life period. Neither tryptamine nor phenylethylamine was detected in the monitored samples; histamine was found only in four cheese samples containing up to 25 mg/kg. The contents of spermine and spermidine were low and did not exceed the values of 35 mg/kg. Significant amounts of tyramine, putrescine, and cadaverine occurred especially in cheeses produced from ewe’s milk or in long-term ripened cheeses. In about 10% of the tested cheeses, the total concentration of all the monitored biogenic amines and polyamines exceeded the level of 200 mg/kg, which can be considered toxicologically significant. In fermented dairy products, the tested biogenic amines occurred in relatively low amounts (generally up to 30 mg/kg) that are regarded safe for the consumer’s health.     

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.     

Screening of biogenic amine production by lactic acid bacteria isolated from grape must and wine

The potential to produce the biogenic amines tyramine, histamine and putrescine, was investigated for lactic acid bacteria (LAB) of various origin, including commercial malolactic starter cultures, type strains and 78 strains isolated from Spanish grape must and wine. The presence of biogenic amines in a decarboxylase synthetic broth was determined by reverse-phase high performance liquid chromatography (RP-HPLC). Tyramine was the main amine formed by the LAB strains investigated. Leuconostoc strains were the most intensive tyramine formers. No potential to form biogenic amines was observed in Oenococcus oeni strains. Two strains of Latobacillus buchneri were associated with putrescine formation. None of the lactic acid bacteria produced histamine. According to these in vitro results, the commercial starter bacteria analyzed did not produce histamine, tyramine and putrescine.     

Content and distribution of biogenic amines in Dutch-type hard cheese

Content of biogenic amines (BA; histamine, tyramine, tryptamine, putrescine, 2-phenylethylamine, cadaverine, spermidine, spermine) and counts of microorganisms (total aerobic and facultative anaerobic mesophilic microorganisms, TAC; lactic acid bacteria, LAB; coliforms; total anaerobic microorganisms, TAN; yeasts + moulds), were evaluated within the ripening interval of 1–6 months in Dutch-type hard cheese, produced by two different producers (the cheeses designated as R30 and R45) using pasteurized milk and different starter cultures (designated as R30-FD and R30-CH in the case of the R30 cheeses). The cheeses from the two producers differed in their fat content (30% and 45% of fat in dry matter, respectively).     

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。     

Content of biologically active polyamines in livers of cattle, pigs and chickens after animal slaughter

Dietary polyamines putrescine (PUT), spermidine (SPD) and spermine (SPM) participate in an array of important human physiological roles, including tumour growth. Physicians and dieticians thus need reliable information on polyamine contents in foods. However, data for livers are lacking. We determined therefore the content of these polyamines 24 h after slaughter in livers of young bulls, cows, pigs and chicken in 58, 19, 36 and 38 samples, respectively. Polyamines were determined as N-benzamides by micellar electrokinetic capillary chromatography. Mean PUT contents about 25 mg kg⁻¹ were found in cattle livers, while very low or negligible contents were determined in livers of the other animals. Extremely high mean SPD contents of 122 and 161 mg kg⁻¹ were found in livers of bulls and cows, respectively and mean levels of 32 and 57 mg kg⁻¹ in livers of pigs and chicken. An opposite relation was observed for SPM. Its mean contents were 43, 35, 115 and 120 mg kg⁻¹ for bulls, cows, pigs and chicken livers, respectively. Thus, livers of the tested animal species belong among foods with the highest polyamine contents. However, the contents ranged very widely, that makes application of the results for the control of human nutrition rather difficult. Polyamine contents in bovine blood were found to be below the detection limits of 2.1, 1.0 and 1.4 mg kg⁻¹ for PUT, SPD and SPM, respectively. Thus, the blood content did not contribute to the substantial polyamine contents in bovine liver found in this study.