Determination of trans‐ and cis‐Urocanic Acid in Relation to Histamine,Putrescine, and Cadaverine Contents in Tuna (Auxis Thazard) at Different Storage Temperatures

Scombroid fish poisoning is usually associated with consumption of fish containing high levels of histamine. However, reports indicate that some cases have responded to antihistamine therapy while ingested histamine levels in these cases were low. Potentiation of histamine toxicity by some biogenic amines, and release of endogenous histamine by other compounds such as cis‐urocanic acid (UCA) are some hypotheses that have been put forth to explain this anomaly. Very little is known about the effects of storage conditions on the production of both UCA isomers and biogenic amines in tuna. Thus, the production of trans‐ and cis‐UCA, histamine, putrescine, and cadaverine in tuna during 15 d of storage at 0, 3, and 10 °C and 2 d storage at ambient temperature were monitored. The initial trans‐ and cis‐UCA contents in fresh tuna were 2.90 and 1.47 mg/kg, respectively, whereas the levels of putrescine and cadaverine were less than 2 mg/kg, and histamine was not detected. The highest levels of trans‐ and cis‐UCA were obtained during 15 d storage at 3 °C (23.74 and 21.79 mg/kg, respectively) while the highest concentrations of histamine (2796 mg/kg), putrescine (220.32 mg/kg) and cadaverine (1045.20 mg/kg) were obtained during storage at room temperature, 10 and 10 °C, respectively. Histamine content increased considerably during storage at 10 °C whereas trans‐ and cis‐UCA contents changed slightly. The initial trans‐UCA content decreased during storage at ambient temperature. Thus, unlike histamine, concentrations of trans‐ and cis‐UCA did not result in elevated levels during storage of tuna.     

Effect of Diamines, Polyamines and Tuna Fish Extracts on the Binding of Histamine to Mucin In Vitro

Binding of [ring-2-¹⁴C]-histamine to mucin was studied. Maximum binding of 2.5 μmol histamine/μmol mucin was observed at a histamine concentration of 0.5 mg/ml. Histamine binding was decreased by 46.5% by spermine, 25.3% by spermidine, 9.5% by cadaverine, and 11.1% by putrescine. These maximum effects were observed at molar ratios of histamine to polyamine of 1:1 for spermine and spermidine, 1:2 for cadaverine, and 1:2.3 for putrescine. The relative effects on histamine binding to mucin of extracts from wholesome canned tuna fish and a mixture of spermine, spermidine, cadaverine and putrescine were also examined. Although the total neutral aqueous extract of tuna had little effect on histamine-mucin binding (5.4% inhibition), the extract of bases from tuna and the prepared base mixture showed maximum inhibition of histamine binding to mucin of 23.1% and 21.4%, respectively. Certain other components of tuna extracts (NaCl, aspartic acid and glutamic acid) did not decrease histamine-mucin binding under the conditions used.     

温度和姜精油对金枪鱼品质影响及生物胺相关性

将分别贮藏于不同温度(0、4、15℃)条件下的金枪鱼样品,以及4℃条件下用姜精油作为生物抗氧化剂处理过的样品作为研究对象,定期测定金枪鱼生物胺(腐胺Put、尸胺Cad、组胺His、酪胺Tyr)、反式尿刊酸的含量、挥发性盐基氮(total volatile basic nitrogen,TVB-N)值、K值和菌落总数,研究温度和姜精油对金枪鱼生物胺含量的影响及组胺与反式尿刊酸的关系。结果表明,温度对金枪鱼生物胺生成有较大影响,0℃条件下生物胺含量显著低于其他贮藏温度,到第6天时组胺、酪胺、腐胺和尸胺含量分别为28.25、5.47、14.84 mg/kg和17.05 mg/kg,而此时15℃条件下金枪鱼组胺、酪胺、腐胺和尸胺含量分别为135.4、14.63、29.49 mg/kg和41.55 mg/kg;生物胺中组胺含量最高,而酪胺含量始终处于低水平状态,即使在15℃条件下到第7天时其含量仍为16.24 mg/kg;结果同时表明,生物抗氧化剂对生物胺的产生和微生物生长有一定的抑制作用,并能有效地延缓蛋白质和ATP降解。相关分析和回归分析表明,组胺、酪胺、尸胺、与TVB-N值、K值和菌落总数高度相关,其相关系数r在0.804~0.981之间,且生物胺和菌落总数、TVB-N值之间存在重要对应关系,反式尿刊酸对组胺的产生影响不大,两者之间相关性较弱,在4℃时其相关系数r为0.630。     

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.     

Quality Indices for Canned Skipjack Tuna: Correlation of Sensory Attributes with Chemical Indices

Different styles of commercially canned skipjack tuna were evaluated by sensory judges as well as by chemical analysis for putrefactive amines as indicators of protein decomposition. Good correlation was found between expert sensory decisions for “accept/reject” quality and levels of putrescine and cadaverine in the product. The study confirmed the repeatability of judgements by trained panelists and the existence of a “cut-off” point for sample acceptance which related to specific levels of putrescine and cadaverine in products which had experienced putrefactive decomposition.     

Effects of on-board and dockside handling on the formation of biogenic amines in mahimahi (Coryphaena hippurus), skipjack tuna (Katsuwonus pelamis), and yellowfin tuna (Thunnus albacares)

Consumer illnesses by scombroid poisonings have been a continuing problem for many years. The intoxications follow the ingestion of fish such as tuna and mahimahi that have undergone bacterial decomposition, leading to the formation of biogenic amines. Research studies have concluded that histamine is one of the indicators of scombrotoxic fish and that other amines, such as cadaverine, could be involved in the illnesses. Guidance for the handling of fish on board fishing vessels to prevent the production of scombrotoxic fish has been limited by a lack of data addressing changes that occur in fish from the water to delivery at dockside. In this study, the changes in selected biogenic amines were determined in mahimahi and tuna, which were captured and held in seawater at 25 to 35 degrees C for incubation times up to 18 h. The fillets from the treated fish were sectioned by transverse cuts and analyzed for histamine, cadaverine, and putrescine. Results showed that at 26 degrees C, more than 12 h of incubation were required before a histamine concentration of 50 ppm was reached in mahimahi. At 35 degrees C, 50 ppm histamine formed within 9 h. Similar results were found for skipjack and yellowfin tuna. Histamine concentrations exceeded 500 ppm within an additional 3 h of incubation in mahimahi. At both temperatures, an increase in the concentration of cadaverine preceded an increase in histamine levels. Changes in putrescine concentrations in the fish were less pronounced. The study also demonstrated that histidine decarboxylase activity was retained in some frozen samples of fish and could result in further increases in histamine on thawing.     

Development of Biogenic Amines in Yellowfin Tuna (Thunnus albacares): Effect of Storage and Correlation with Decarboxylase-Positive Bacterial Flora

ABSTRACT: The effects of storage at 0,4,10, and 22°C for 0,1,3,5, and 9 d on the quality of yellowfin tuna fillets as determined by microbiological assessment, development of some biogenic amines, and sensory analysis were studied. Tuna fillets stored at 22 °C for 3 d, 10 °C for 5 d, and 4 °C for 9 d were rated unacceptable for consumption. Those stored at 22 °C for 3 d had total aerobic bacterial count of > 8 log₁₀ CFU/g, a histamine-producing bacterial population of 7 log₁₀ CFU/g, and 832 ppm of histamine, 35.8 ppm of putrescine, and 147 ppm of cadaverine. A comparison of the capillary electrophoresis, AOAC fluorometric method, and gas chromatography showed a very good correlation (r² > 0.99) among these 3 methods for histamine quantitation in tuna samples. Morganella morganii, Enterobacter agglomerans, Enterobacter intermedium, Pseudomonas fluorescens, Proteins vulgaris , and Serratia liquefaciens were the decarboxylase-positive bacterial species isolated by using the Niven's medium and identified during storage, which were responsible for histamine production in test tuna fillets.     

Biogenic amine production by<Emphasis Type="Italic"> Morganella morganii</Emphasis> and<Emphasis Type="Italic"> Klebsiella oxytoca</Emphasis> in tuna

Biogenic amine formation in tuna inoculated with two micro-organisms reported as powerful in vitro histamine producers, Morganella morganii and Klebsiella oxytoca, was investigated. Inoculated and non-inoculated tuna samples were stored in ice, under refrigeration at 8–10 °C and at 20±1 °C. Ten biogenic amines were studied and changes were found only in cadaverine and histamine in inoculated samples. Therefore, the in vitro ability of K. oxytoca to form cadaverine and that of both microorganisms to form histamine was confirmed in inoculated samples. Storage temperature had a crucial effect on the amount and intensity of amine production. On the other hand, the potential ability of M. morganii to form putrescine was not shown in the inoculated samples, regardless of the storage temperature.     

Determination of biogenic amines in fresh unpacked and vacuum-packed beef during storage at 4°C

Biogenic amines, putrescine, cadaverine, and histamine were determined in unpacked and vacuum-packed beef; during storage at 4°C for 12 and 35 days, respectively, under the same conditions. Analysis by High Performance Liquid Chromatography in both kind of samples, showed variation of cadaverine and histamine with an increasing tendency to a definite increase of putrescine. In unpacked samples, maximum mean contents for each amine reached 10.4, 5.2 μg/g for putrescine and cadaverine, respectively (12th day) and 2.2 μg/g for histamine (5th day). The total amine content reached 16.5 μg/g (12th day). After 8 days of storage, organoleptic fall-off began and, on the 12th day, all samples were disqualified. In vacuum packs, amine levels increased after 19 days. Maximum levels were determined for putrescine on the 35th day (36.3 μg/g), for cadaverine on the 26th day (28.9 μg/g) and for histamine on the 35th day (19.0 μg/g). Although the vacuum-packed beef was organoleptically acceptable up to 35 days, the total amine levels were greater than 5 μg/g after the 12th day, indicating a leveling off of quality.     

Amino acid composition of whole mackerel (Scomber scombrus) stored anaerobically at 20°C and at 2°C

Total amino acid composition of whole mackerel ( Scomber scombrus ) and of milt and roe from mackerel stored at different conditions were studied. During storage at low temperature, no obvious changes were found. When stored at high temperature (20°C), the contents of several of the amino acids decreased, and phenylethylamine, tyramine, putrescine, histamine and cadaverine were formed the formation of cadaverine was higher than the loss of lysine.     

Biogenic Amine Content of Beers Consumed in Turkey and Influence of Storage Conditions on Biogenic Amine Formation

The biogenic amine content of beers available in Turkey, both domestic production and imported products, was investigated. A total of 17 domestic and 13 imported beers were evaluated in terms of 8 different biogenic amines (histamine, tyramine, spermine, spermidine, 2‐phenylethylamine, putrescine, tryptamine and cadaverine). HPLC methodology with pre‐column derivatization and photodiode array detection after derivatization with dansyl chloride was used for quantification. In addition, the evolution of important amines such as histamine, tyramine, putrescine and cadaverine were investigated during different storage conditions by response surface methodology. The results indicated that both storage temperature and storage time were important factors affecting biogenic amine content. Histamine and putrescine increased during storage, but then decreased after reaching a maximum level after six weeks. With the biogenic amines tyramine and cadaverine, the amounts increased during the entire storage period. At higher storage temperatures, the formation of biogenic amines increased.     

Effects of Storage Time and Temperature on the Microflora and Amine-Development in Spanish Mackerel (Scomberomorus maculatus)

Microbial content was characterized and levels of three amines (histamine, cadaverine, and putrescine) were determined in Spanish mackerel (Scomberomorus maculatus) decomposed at PC, WC, and 30°C for varying lengths of time. Correlations were shown (1) between the levels of the histamine, cadaverine, and putrescine and the time and temperature of decomposition, (2) between the ratios of cadaverine/histamine and putrescine/histamine levels and the temperature of decomposition, and (3) between increasing total microbial counts and rising amine levels. A total of 14 bacterial species with histidine decarboxylase activity were isolated from decomposing fish, including three species (Acinetobacter lwoffi, Pseudomonas putrefaciens, and Aeromonas hydrophila) not previously reported to have the potential to produce histamine.     

Free Amino Acids in Dark- and White-muscle Fish as Determined by O-phthaldialdehyde Precolumn Derivatization

ABSTRACT: Free amino acids in fish reflect microbial spoilage, and are precursors of biogenic amines, factors of health concern and indicators offish decomposition. The objective of this research was to quantify levels of free histidine, lysine, ornithine and glutamine, which may become originators of highly undesirable histamine, cadaverine and putrescine in fish tissue. Liquid chromatography using o-phthaldialdehyde (OPA) pre-column derivatization, gradient elution and a C18 column were used for separation. Histidine was higher in white tissue of mahi-mahi and tuna than in red tissue. No significant difference in lysine levels was found between the two tissues of tuna. Glutamine and ornithine were higher in red tissue of mahi-mahi and tuna. Red snapper had higher levels of lysine than histidine.     

Histamine and biogenic amine production by Morganella morganii isolated from temperature-abused albacore

Histamine-producing bacteria were isolated from albacore stored at 0, 25, 30, and 37 degrees C. They were screened using Niven's differential medium, and their histamine production was confirmed by high-pressure liquid chromatography analysis. The optimum temperature for growth of histamine-producing bacteria was 25 degrees C. The bacterium producing the highest level of histamine was isolated from fish abused at 25 degrees C. It was identified as Morganella morganii by morphological, cultural, biochemical, and antimicrobial characteristics and by the Vitek microbial identification system. The M. morganii isolate was inoculated into tuna fish infusion broth medium, and the effect of temperature was determined for microbial growth and formation of histamine and other biogenic amines. The isolate produced the highest level of histamine, 5,253 ppm, at 25 degrees C in the stationary phase. At 15 degrees C, histamine production was reduced to 2,769 ppm. Neither microbial growth nor histamine formation was detected at 4 degrees C. To determine whether the isolate can also produce other biogenic amines that can potentiate histamine toxicity, production of cadaverine, putrescine, serotonin, tryptamine, tyramine, phenylethylamine, spermidine, and spermine by the isolate was also monitored. Cadaverine, putrescine, and phenylethylamine were detected with microbial growth in the tuna fish infusion broth medium. The optimum temperature for cadaverine, putrescine, and phenylethylamine formation was found to be 25 degrees C, as it was for histamine.     

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.     

抗氧化剂对金枪鱼肉冻藏过程中组胺的抑制作用

利用C18柱、紫外检测器的高效液相色谱建立快速准确检测金枪鱼鱼肉中组胺含量的方法和技术,并对抗氧化剂对金枪鱼储藏过程中组胺产生的抑制作用进行研究。对不同部位的鱼肉采用不同抗氧化剂浸泡处理,检测不同储藏时间下组胺含量的产生和变化。结果表明：该方法的线性回归方程为Y=5.3748X＋11.42,线性范围为10～1000μg/mL,相关系数为0.9986,加标回收率为92.0%～111.3%,检出限为3.684μg/mL,用于实际样品测定操作简单、准确灵敏、适用性强、结果满意;经抗氧化剂处理的金枪鱼鱼肉中组胺含量明显低于未经处理的,其中异VC钠和柠檬酸钠混合溶液的浸泡效果最好、VC钠次之、柠檬酸钠最差。抗氧化剂既可以防止金枪鱼鱼肉氧化变黑,又可有效降低组胺的产生。     

Bacterial Growth and Histamine Production on Vacuum Packaged Tuna

Vacuum and nonvacuum packaged samples of tuna or tuna spiked with Klebsiella oxytoca T₂, Morganella morganii JM or Hafnia alvei T₈ were stored at 2°C and 10°C and examined for bacterial growth and histamine production on day 3, 6, 10 and 15. Unlike those stored at 2°C, tuna samples stored at 100°C showed significant bacterial growth over time and had high levels of histamine (>200 mg/100g tuna on day 15). Vacuum packaging did not show any beneficial effect in controlling bacterial growth and histamine production on tuna samples at these two temperatures. Low temperature storage was more effective than vacuum packaging in controlling histamine production by the bacteria tested.     

A rapid dipstick test for histamine in tuna

This study describes the production of a solid phase assay (dipstick test) for histamine in tuna based on the coupling of diamine oxidase to a peroxidase/dye system. The optimized dipstick preparation consisted of a diamine oxidase/peroxidase/Stabilcoat mixture layered onto a tetramethylbenzidine color strip. The assay was linear to 1.0 mM histamine (RSQ = 0.994) and the minimum detectable concentration was 0.07 mM corresponding to 2.3 mg% in tuna extracts. Quality control samples gave intra and interassay precisions of 9.14–9.84 %CV and 7.08–13.86 %CV respectively. Similarly, the assay was accurate (p> 0.05) for both low and high controls. Histidine and cadaverine did not interfere with the assay whereas putrescine reacted slightly with the dipstick. Histamine determinations of fresh and spoiled tuna gave good agreement between the dipstick method and both the official reference and modified AOAC flurometric methods.     

Formation of biogenic amines in herring and mackerel

The formation of biogenic amines (histamine, cadaverine, putrescine and spermidine) was followed during vacuum packed storage at 2 degrees C or 10 degrees C in the scombroid fish mackerel and in the non-scombroid fish herring. Also the changes in the content of free amino acids and in the organoleptic and microbiological qualities were followed. At 10 degrees C the amine contents were 2-20 times higher at the time of rejection as compared with samples stored at 2 degrees C. In herring and mackerel similar amounts of histamine were accumulated, whilst cadaverine was formed at much higher levels in mackerel compared with herring. The high contents of cadaverine in mackerel can possibly explain why mackerel and not herring are often implicated in incidents of scombrotoxic poisoning.     

冷藏期内猪肉肌苷酸与生物胺两者变化关系的研究

采用高效液相色谱法测定生鲜猪肉的背最长肌在4℃温度条件下肌苷酸(IMP)和腐胺、尸胺、组胺、酪胺、亚精胺、精胺这六种生物胺含量的变化情况,从而进一步分析它们之间的相关性。结果表明,随着货架期的延长,IMP含量呈先上升后降低的趋势,并在第2d达到最高;精胺含量基本保持在4.0mg/kg左右,组胺的含量始终很低,其它的胺类物质都有所增加,尸胺的变化最为突出;虽然腐胺和亚精胺的含量比较低,但是仍然有明显的变化;酪胺变化也非常明显。从Person积差相关系数可以看出,IMP与其他指标相关系数呈负相关显著,有的指标是不显著的;在0.01水平上,IMP与亚精胺之间的负相关性最强,达到了-0.981;其次是尸胺与IMP之间,相关系数是-0.960,呈显著负相关;酪胺也与IMP显著负相关;IMP与腐胺、组胺、精胺的相关系数都不显著。因此,通过测定IMP含量变化可以预测猪肉新鲜度,且可作为猪肉保藏与加工过程中品质控制的重要指标之一。