Effects of Modified Atmosphere Packaging with Various Carbon Dioxide Composition on Biogenic Amines Formation in Indian Mackerel (Rastrelliger kanagurta) stored at 5 ± 1°C

The profile of major biogenic amines was investigated in Indian mackerel packed in modified atmosphere for up to 12 days at 5 ± 1°C. Beheaded and gutted Indian mackerel was packed under different carbon dioxide compositions to study the effects on biogenic amines formation. The treatments were control air (C), vacuum packaging (VP), 30% CO₂/65% N₂/5% O₂ (M30C), 60% CO₂/35% N₂/5% O₂ (M60C), 80% CO₂/15% N₂/5% O₂ (M80C) and 100% CO₂ (M100C). Each amine responded differently to different CO₂ levels. After 12 days of storage, concentrations of histamine were reduced by 6.4%, 8.5%, 70.3%, 78.8% and 90.2% in fish packed under VP, M30C, M60C, M80C and M100C, respectively as compared with control air. Changes in putrescine and cadaverine showed a similar pattern. Gas mix of M30C and VP stimulated the formation of tyramine reaching 203 and 172 ppm, respectively. Higher composition of CO₂ had a significant inhibitory effect on tyramine concentration (p < 0.05). There were parallel increases of putrescine and spermidine in C, VP and M30C. No significant effect of CO₂ was observed on spermine (p > 0.05). After 9 days of storage, more than 300 ppm of histamine was detected in mackerel packed in VP and M30C; therefore, these atmospheres pose a histamine intoxication risk. Copyright © 2013 John Wiley & Sons, Ltd.     

赖氨酸脱羧酶发酵工艺及酶学性质

考察了蜂房哈夫尼菌(H.alveiAS1.1009)的L-赖氨酸脱羧酶对L-赖氨酸的脱羧作用,分析了培养基、温度、pH、激活剂等因素对酶活力的影响。实验结果表明,最适培养基成分为:ρ(蔗糖)=20 g/L、ρ(玉米浆)=20g/L、ρ(酵母膏)=5 g/L、ρ(牛肉膏)=3 g/L、ρ(蛋白胨)=10 g/L、ρ(氯化钠)=5 g/L、ρ(硫酸铵)=2 g/L、ρ(维生素B6)=5 g/L和ρ(L-赖氨酸)=5 g/L,100 mL发酵液中接种量为5 mL液体菌种。最佳转化工艺条件为:转化体系温度35℃、pH=5.0,ρ(菌体)=10 g/L,ρ(吐温-80)=0.15 g/L。L-赖氨酸脱羧酶在最适发酵和转化条件下比酶活可达7 984.43 U,底物转化率可达70%。     

Stimulation of Cadaverine Production by Foodborne Pathogens in the Presence of Lactobacillus,Lactococcus, and Streptococcus spp.

Abstract: The effect of Lactobacillus plantarum (FI8595), Lactococcus lactis subsp. cremoris MG 1363), Lactococcus lactis subsp. lactis (IL 1403), and Streptococcus thermophilus on cadaverine and other biogenic amine production by foodborne pathogens was investigated lysine decarboxylase broth. Both of lactic acid bacteria and foodborne pathogens used (especially Staphylococcus aureus, E. coli, Lc. lactis subsp. lactis and Lb. plantarum) had an ability to convert aminoacids into biogenic amine. The conversion of lysine into cadaverine was the highest (167.11 mg/L) by Lactobacillus spp. Gram‐positive bacteria generally had a greater ability to produce cadaverine with corresponding value of 46.26, 53.76, and 154.54 mg/L for Enterococcus faecalis, S. aureus, and Listeria monocytogenes, respectively. Significant variations on biogenic amine production were observed in the presence of lactic acid bacteria strains (P < 0.05). The role of lactic acid bacteria on biogenic amine production by foodborne pathogens varied depending on strains and specific amine. Cadaverine accumulation by Enterobactericeae was increased in the presence of lactic acid bacteria strains except for St. thermophilus, which induced 2‐fold lower cadaverine production by S. Paratyphi A. Lc. lactis subsp. lactis and Lc. lactis subsp. cremoris induced 10‐fold higher increases in histamine for E. coli and K. pneumoniae, respectively. Lactic acid bacteria resulted in strong increases in cadaverine production by P. aeruginosa, although remarkable decreases were observed for histamine, spermidine, dopamine, agmatine, and TMA in the presence of lactic acid bacteria in lysine decarboxylase broth . The result of the study showed that amine positive lactic acid bacteria strains in fermented food led to significant amine accumulation by contaminant bacteria and their accumulation in food product may be controlled by the use of proper starters with amine‐negative activity. Practical Application: Foodborne pathogens and certain lactic acid bacteria are particularly active in the production of biogenic amines. Most of the strains of bacteria possess more than 1 amino acid decarboxylase activity under lysine enrichment culture conditions. Lactic acid bacteria strains had a significant role on increase putrescine accumulation by foodborne pathogens. The increased production of biogenic amines in mixed culture is the result of presence of amine positive lactic acid bacteria strains. The addition of a proper selected starter culture with amine‐negative activity is advisable to produce safer fermented food with low contents of biogenic amines.     

屠宰过程中微生物污染与冷却肉品质的关系研究

研究了微生物污染的数量与各品质指标的变化关系,并对剥皮与烫毛两条生产线控制前后的初始微生物与各品质指标进行了对比研究。结果表明：冷却肉的品质变化与微生物的生长繁殖程度紧密相关,当微生物数量达到108 cfu／g时,产品已经开始腐败;通过对冷却肉中的微生物数量与冷却肉中尸胺含量进行回归分析,得出回归方程为：微生物数量＝5．145＋0．557×尸胺（R＝0．959,R2＝0．919,Adj． R2＝0．879）;对是否采用控制措施的起始微生物数量及相关肉品质指标进行测定,采用控制措施显著地减少了初始微生物数量,使得冷却肉的保鲜期（0～4℃条件贮藏）延长3～4d。     

Biogenic Amines Formation in Bigeye Tuna Steaks and Whole Skipjack Tuna

ABSTRACT: Putrescine, cadaverine and histamine formation was monitored in fresh and frozen/thawed Bigeye tuna steaks ( Thunnus obesus ) and whole Skipjack ( Katsuwonus pelamis ) stored at room temperature and in ice and to determine the potential use of putrescine and cadaverine as indicators of decomposition. While all biogenic amines were formed during both storage conditions, putrescine was formed at a much lower rate and concentration. For both storage conditions and species, cadaverine appeared prior to and/or accumulated at a faster rate than histamine. Skipjack tuna had a faster accumulation of cadaverine than Bigeye under the same storage conditions. The results indicated that cadaverine, either alone or with histamine, could be used as an index of decomposition for Skipjack and Bigeye tuna.     

荧光法测定微生物转谷氨酰胺酶的探讨

讨论了荧光法测定微生物ＴＧａｓｅ的条件，对反应体系中ＭＤＣ的浓度、反应时间、ＤＴＴ的浓度、硫铵的浓度、酶浓度等因素与体系荧光强度的关系进行了探讨，得出３０μｍｏｌ／ＬＭＤＣ１．０ｍＬ，２．２９ｍｇ／ｍＬＤＭＣ１．０ｍＬ，５．５ｍｍｏｌ／ＬＤＴＴ０．３ｍＬ，加入一定量酶样品，３７℃反应３０ｍｉｎ后用１ｍｏｌ／Ｌ硫铵溶液０１ｍＬ灭活，测定体系在３６０ｎｍ激发光下，５３０ｎｍ处的荧光强度值，以此作为定量的依据。     

Concentrations of biogenic amines in fish,squid and octopus and their changes during storage

The concentrations of seven biogenic amines (BA) were simultaneously determined in 74 samples of fish, squid and octopus, by the method of HPLC coupled with pre-column derivatisation. The relationship between the formation of BA in aquatic products and the growth of microbial flora during storage was also investigated. Results showed that putrescine, cadaverine, histamine and tyramine were the dominant BA in the studied samples, but the concentrations of histamine and tyramine were mostly less than 50 and 100 mg kg⁻¹, respectively. Freezing can effectively prevent the formation of BA, but the levels of putrescine, cadaverine, histamine and tyramine significantly increased (p < 0.05) during storage at 4 and 25 °C. The growth of mesophilic or psychrophilic bacteria in blue scad and octopus strongly and positively correlated with the formation of amines (such as putrescine, cadaverine, histamine and tyramine) during storage, except for histamine in octopus.     

Amperometric Biosensor for Total Histamine, Putrescine and Cadaverine using Diamine Oxidase

Diamine oxidase was purified from porcine kidney to homogeneity and immobilized onto a porous preactivated nylon membrane. The enzymic membrane was attached to an amperometric electrode for determination of the total concentrations of histamine, cadaverine and putrescine accumulated in fish fillets during storage. For detecting hydrogen peroxide released, the working platinum electrode was poised at +400 mV vs Ag/AgCl to prevent electrode fouling as well as minimize electroactive interferences. The biosensor was linear up to 6 mM with a lower detection limit of 25 μM for the three substrates and steady state response was achieved within 1.5 min. The enzyme membranes were stable at 5°C for 2 mo and usable for at least 60 assays.     

Monitoring of beef aging using a two-line flow injection analysis biosensor consisting of putrescine and xanthine electrodes

A two-line flow injection analysis (FIA) biosensor was developed which can simultaneously detect bacterial spoilage and the progress of aging. This FIA biosensor was composed of a putrescine oxidase immobilized electrode and a xanthine oxidase immobilized electrode as detectors. The putrescine electrode measures putrescine and cadaverine which are produced by bacteria, and the xanthine electrode measures hypoxanthine and xanthine which accumulate in meat with aging.

The analytical conditions for this system were set as follows; flow rate, 1 ml/ min; water bath temperature, 30 °C; flow buffer, 0.1 M phosphate buffer (pH 7.0); injection volume for putrescine electrode, 200 μl; injection volume for xanthine electrode, 40 μl; and measurement cycle, 2 min.

The linear relationship for standard solution was between 20 and 800 nmol/ml in the putrescine electrode and between 0.1 and 3.0 μmol/ml in the xanthine electrode. The coefficients of variation in standard solutions were 2.14% with the putrescine electrode and 2.83% with the xanthine electrode. The coefficients of variation values in the specimen solution were 3.22% and 3.76%, respectively.

This two-line FIA biosensor was applied to the vacuum-packed beef stored at 0, 5 and 10 °C. The progress of aging could be monitored at all temperatures, and the bacterial spoilage could be detected before the appearance of putrid odor at 5 and 10 °C. However, at 0 °C the putrid odor did not appear during storage, and neither putrescine nor cadaverine was detected. Thus, this FIA biosensor was confirmed to be useful for the quality control of beef aging at 5 and 10 °C, but not at 0 °C.