Chemometrics Approaches to Monitoring of Biogenic Amines Changes in Three Fish Species

The biogenic amine contents in fillets of three freshwater fish of kutum (Rutilus frisii), common carp (Cyprinus carpio), and Caspian salmon (Salmon caspius) were monitored during storage at 4°C for 20 days. The amounts of biogenic amines were determined at regular intervals using high performance liquid chromatography after derivatization with benzoyl chloride. Significant differences in the levels of selected biogenic amines among the fish samples were found (p < 0.05) during storage times. While the amount of putrescine, cadaverine, histamine, and tyramine in the fish fillets increased during the storage time, the changes in spermine and spermidine contents of the fish species were minor and both showed low and inverse correlations with storage time. For each of the three fish species, the sensory acceptability limit was found to be 4–8 days. The quality and biogenic amine indices of fish samples showed high correlation with storage time (r > 0.9). The principal component analysis of the results indicated that the formation of biogenic amines displays a certain behavioral pattern in all fish samples.     

草鱼冷藏过程鱼肉品质与生物胺的变化及热处理对生物胺的影响

文章研究了草鱼（Ctenpharyngodon idellus）冷藏过程鱼肉品质与生物胺的变化及热处理对生物胺的影响.将经过前处理的草鱼鱼片于4℃条件下贮藏,对其感官品质、挥发性盐基氮（TVB-N）、菌落总数、汁液流失率以及生物胺等指标进行测定,另外经过85℃、15 min的热处理后对草鱼鱼片的生物胺进行测定,探求其变化规律及草鱼的生物胺与其品质指标的相关性.结果表明,随着贮藏时间的延长,草鱼的汁液流失率、TVB-N、菌落总数均呈现明显的上升趋势,与感官分值的变化趋势相符,草鱼的感官接受极限为9d.草鱼鱼片在冷藏加工过程中变化最显著的生物胺是腐胺和尸胺,第15天腐胺的质量分数达到（17.12±4.40） mg·kg^-1,尸胺的质量分数达到（237.47 ±3.96） mg·kg^-1,且腐胺和尸胺与其他评价指标有着较好的相关性.经热处理后,腐胺、尸胺、组胺、精胺的质量分数有所下降,但与未经热处理的鱼片没有显著性差异（P＞0.05）,故85℃、15 min的热处理对于草鱼中生物胺的破坏作用不明显,难以有效地去除生物胺.     

Formation of Biogenic Amines in Delayed Salt-Cured Whole and Gutted Indian Oil Sardines (Sardinella longiceps)

The formation of biogenic amines in Indian oil sardines (Sardinella longiceps) collected from the Tuticorin coast of South India and treated with delayed salt-curing was investigated. Sardines were wet salt-cured in whole and gutted forms and examined in fresh and after 6, 9, 12, and 15 h delayed conditions at ambient temperature (32 ± 2°C) at four stages of salt-curing process for quality parameters and formation of biogenic amines. Moisture content decreased from 76% to 11.3% in salting followed by drying. Similarly, water activity reduced from 0.96 to 0.74 in salt-cured sardines. In contrast, total volatile base nitrogen content increased from 79 to 3,590 mg kg^–1 in salt-cured sardines. Halophilic count of sardines was higher (8 log cfu g^–1) after salting and decreased in drying stages. The 15 h delayed salt-cured whole sardines had higher histamine contents (1,568.27 mg kg^–1), cadaverine (4,059.84 mg kg^–1), putrescine (1,604.95 mg kg^–1), and tyramine (862.94 mg kg^–1) on the final day of drying. Whole sardines had higher biogenic amines and halophilic bacterial counts than the gutted sardines. The whole and gutted sardines salt-cured up to a delay of 6 and 12 h, respectively, at ambient temperature yielded the acceptable quality products with respect to biogenic amines.     

高效液相色谱法测定生物胺衍生条件的优化研究

利用荧光检测器（FLD）,采用柱前衍生高效液相色谱法（HPLC）对组胺（HIS）、色胺（TRP）、2-苯乙胺（2-PHE）、腐胺（PUT）、尸胺（CAD）、酪胺（TYR）、亚精胺（SPD）和精胺（SPM）8种生物胺（BA）进行测定分析。通过探究丹酰氯（Dns-Cl）质量浓度、反应体系的pH、衍生时间及衍生温度对生物胺衍生反应的影响,确定最优的衍生条件。结果显示,当Dns-Cl的质量浓度为BA质量浓度的10倍、pH 11.0、40℃下避光反应45 min,所有BA均能有效分离。BA的相对标准偏差（RSD）在3%以内,能够满足分析要求。     

Effect of Bacterial Community and Free Amino Acids on the Content of Biogenic Amines During Fermentation of Yu-lu,a Chinese Fermented Fish Sauce

The main objective of this work was to investigate the effects of bacterial community and free amino acids on the content of biogenic amines in Yu-lu during fermentation. Four major biogenic amines (histamine, tyramine, cadaverine, and putrescine) were identified, and the content of tyramine was positively correlated with histamine (R value = 0.9113). Aspartic acid, glutamic acid, histidine, leucine, and lysine were the dominant free amino acids. 16S rRNA sequencing showed that the composition of the bacterial community changed significantly during fermentation of Yu-lu. Principal component analysis revealed the crucial links between microbial community and biogenic amines. For example, Halanaerobium was probably associated with the formation of putrescine, while Halomonas might be associated with the degradation of biogenic amines at the end of fermentation of Yu-lu. This study provided a detailed evaluation of the Yu-lu fermentation process, enabling development of better strategies for biogenic amine control in fish sauce.     

Biochemical and Microbiological Assessment of Crucifix Crab (Charybdis feriatus) Stored at 4°C

Charybdis feriatus is a highly prized crab in the local and international markets of East Asian countries. In local markets, crabs are sold live, iced, or refrigerated at 2°C – 4°C. The present study was carried out to determine the shelf life of whole crab stored at 4°C for 192 h using a set of biochemical, microbiological, and sensorial parameters. Biochemical indicators such as the total volatile base nitrogen (TVB-N) and trimethylamine nitrogen (TMA-N) increased significantly during storage and within an acceptable limit up to 144 h. Regarding the biogenic amines, cadaverine and spermidine increased significantly (p < 0.05) during the storage period. Among them, cadaverine could be proposed as a useful spoilage indicator for this crab species. The mesophilic and psychotropic bacterial count exceeded the limit of acceptance after 144 h of storage. Sensory attributes were gradually diminished with the storage time. Sensory scores were found to correlate well with both mesophiles and psychrotrophs. Based on the results of this study, the meat quality of Charybdis feriatus can be maintained up to 144 h when properly stored at a temperature of 4°C ± 1°C.     

Effectiveness of Synthetic Molecules,and Animal and Vegetable Extracts as Baits for Harvesting Red Swamp Crayfish,Procambarus clarkii

Abstract

The present study was designed to evaluate the efficiency of different biogenic amines (putrescine, cadaverine, tyramine, spermine and spermidine) and their precursory amino acids (arginine, histidine, lysine and tyrosine), as well as animal extracts (from fish, crustaceans and mollusks) and vegetable extracts (Chara sp., coconut and alfalfa) as baits in traps for red swamp crayfish, P. clarkii. The methodology was conceived as a logical sequence to proceed from rapid screening of a large number of treatments and was divided into three phases: (1) chemo-detection bioassays, (2) chemattraction bioassays, and (3) field bioassays carried out under natural conditions. The attractants that showed the best performance as baits for P. clarkii, were: the fish soluble extract (included at 2.96%), the putrescine (included at 0.30%) and the red crab soluble extract (included at 2.69%). The benefits of using these attractants, when added to common pelleted feed, come from their low cost, availability and enhanced handling properties.     

Changes in Biogenic Amines and Bacteria of Tiger-Toothed Croaker (Otolithes ruber) during Ice Storage

Changes in the content of biogenic amines (BA; e.g., putrescine, tyramine, and histamine) and the mesophilic and psychrotrophic bacterial counts of tiger-tooth croaker (Otolithes ruber) were monitored during ice storage for 18 days. BA and bacterial loads gradually increased (p < 0.05) with storage time. On the first day of storage, no amines were detected, and no histamine was detected up to 6 days. The best linear regressions between BA and bacterial counts were found between putrescine and psychrotrophic bacteria. Furthermore, it was found that psychrotrophs were the dominant microorganisms during storage and showed significant (p < 0.05) differences with storage time. The initial concentration of putrescine was 1.27 μg g^?1 and reached 18.8 μg g^?1 when the psychrotrophic bacterial load was over 10⁸ CFU g^?1. The results suggest that the level of putrescine is a suitable indicator of freshness for O. ruber, and the shelf life of O. ruber stored in ice as determined by BA content and microbiological data was 10 days.     

Physicochemical and Microbiological Changes During Drying of Wolf Herring (Chirocentrus dorab) and Coastal Trevally (Carangoides coeruleopinnatus)

The aim of the present study was to assess the physicochemical and microbiological changes during sun drying of salted wolf herring (Chirocentrus dorab) and coastal trevally (Carangoides coeruleopinnatus). For that purpose, the pH value, moisture, sodium chloride (NaCl) content, total volatile base nitrogen (TVB-N), histamine, cadaverine, putrescine, tryptamine, tyramine, spermine, and spermidine, as well as total aerobic mesophilic count, amine forming bacteria, total coliforms, Staphylococcus spp., and Bacillus spp., were determined. The initial pH value was 6.4 and increased during the salt drying process to 6.9 in both cases. The initial moisture, salt, and TVB-N levels of C. dorab and C. coeruleopinnatus were 64.3 and 60.3%, 2.55 and 2.70%, and 22.8 and 16.2 mg/100 g, respectively. At the end of drying, moisture decreased to 31.3 and 35.6%, respectively; salt increased to 13.71 and 16.04%; and TVB-N increased to 35.9 and 33.13 mg/100 g, respectively. Regarding total aerobic mesophilic count, amine forming bacteria, total coliforms, Staphylococcus spp., and Bacillus spp., a statistically significant (P < 0.05) reduction of the population was observed in both cases. Regarding the biogenic amine forming bacteria, Morganella morganii and Klebsiella pneumoniae were isolated from C. dorab; Bacillus cereus, Staphylococcus xylosus, and Providencia rettgeri were isolated from C. coeruleopinnatus. During sun drying, the amount of histamine, cadaverine, putrescine, and tryptamine was reduced; spermine was detected in C. dorab only during the first day, whereas spermidine was not detected. This reduction may be attributed to the presence of biogenic amine decomposing bacteria. However, further research is necessary in order to verify in situ this capacity and exploit potential applications for fish and fishery products.     

The Effects of Fermentation Process with Acid and Lactic Acid Bacteria Strains on the Biogenic Amine Formation of Wet and Spray-Dried Fish Silages of Discards

The impacts of fermentation process with acid and lactic acid bacteria strains (Lactobacillus plantarum, Lactobacillus brevis, Pediococcus acidilactici, Enterococcus gallinarum, and Streptococcus spp.) on the biogenic amine formation of wet and spray-dried fish silage obtained from whole gibel carp (Carassius gibelio, freshwater discard), whole ponyfish (Equulites klunzingeri, seawater discard), and processing by-products of seabass (Dicentrarchus labrax) were investigated. The results showed that among biogenic amines, cadaverine, putrescine, spermidine, spermine, serotonin, dopamine, and agmatine were predominant in all groups. Significant differences (p < 0.05) in biogenic amine concentrations of wet and spray-dried fish silage were observed. Raw fish and wet silages contained histamine level lower than the allowable limit of 50mg/kg, indicating the use of raw fish material with low microbial counts. In addition, no histamine was detected in spray-dried fish silage, except for seabass by-products with a trace quantity of histamine (<0.04mg/100g). The starter culture used for silage preparation did not effectively retard formation of biogenic amines compared to acid silage. It can be concluded that there is potential use of fermented fish silage as a protein source and possibly as a probiotic ingredient for animal feed in both wet and dry form.     

The use of biochemical,sensorial and chromaticity attributes as indicators of postmortem changes in commercial‐size,cultured red porgy Pagrus pagrus,stored on ice

The freshness of red porgy slaughtered in ice slurry and stored in melting ice was evaluated instrumentally, biochemically and sensorialy. Additionally, postmortem skin colour changes were monitored, in an attempt to demonstrate the use of chromaticity parameters as a reliable and convenient approach to quality assessment. Dielectric properties showed a statistically significant decrease (P<0.05) from 12±0 on day 0 to 7.85±0.31 on day 7; there was also a significant decrease in muscle polyamines with more than two amino groups (spermidine and spermine) and an increase in the di‐amine putrescine. The sensorial score was significantly decreased from 30±0 to 4.33±0.21 on day 7. Minimal lightness (L^*) and hue (H°ab) at the dorsal skin area were observed on days 1 and 3 following harvesting. Finally, there was a marked decrease in the entire colour index (ECI – a combination of skin hue and chroma) of both the dorsal and the ventral area in day 3 onwards to day 7, as well as a statistically significant correlation between all the estimated freshness indices and ECI. Based on all these, we inferred that the total polyamines (or putrescine to spermidine and spermine ratio) and ECI could be reliable estimators of freshness, at least under the experimental conditions applied.     

Quality Indices of Cooked Southern King Crab (Lithodes santolla Molina, 1782) Meat During Storage at 0 and −20°C

ABSTRACT

The aim of this work was to determine the changes in quality of cooked southern king crab (Lithodes santolla) meat during storage at 0°C for 5 and 10 days and ?20°C for 10, 30, 60, and 90 days. Chemical indices varied from the initial to the final storage time in refrigerated and frozen samples as follows: TVB-N, 15.8–29.2 and 15.8–23.2 mg/100 g; TMA, 0.0–8.3 and 0.0–1.2 mg/100 g; formaldehyde, 0.5–8.6 and 0.5–3.7 mg/kg; indole, 3.3–20.7 and 3.3–6.6 µg/100 g; lactic acid, 16.4–39.1 and 16.4–16.6 mg/100 g; and P ratio, 0–28.4 and 0–5.2%, respectively. The total nucleotide content rendered 6.77 µmol/g, with adenosine monophosphate (AMP) the main component. The nucleotide indices varied in refrigerated and frozen samples as follows: K, 7.37–27.5 and 7.37–10.3%; Ki, 58.3–86.5 and 58.3–82.7%; H, 30.2–54.9 and 30.2–38.0%; G, 19.4–57.8 and 19.4–15.8%; P, 17.2–42.3 and 17.2–14.7%; and Fr, 41.6–13.4 and 41.6–17.3%, respectively. Biogenic amines found at both storage temperatures were spermine, spermidine, tyramine, and agmatine. Spermine may be considered as a potential freshness index for refrigerated samples and spermidine for frozen storage. The tyramine level should be monitored. During the storage period tested, none of the chemical indices studied exceeded the legal limits.     

Histamine content and isolation of histamine-forming bacteria in fish meal and fish soluble concentrate

Forty fish meals and five fish soluble concentrates (FSC) imported to Taiwan were sampled to determine biogenic amines and histamine-forming bacteria. Average levels of water activity (0.8), water content (39.9%), salt content (7.32%), and total volatile basic nitrogen (TVBN) (150.7 mg/100 g) in FSC samples were significantly higher than those of fish meal samples, whereas the average levels of crude fat (3.94%) and crude protein (40.7%) in FSC samples were lower than those of fish meal samples. Thirty-five (87.5%) fish meal samples and 5 (100%) FSC samples had TVBN levels above the decomposition limit level of 30 mg/100 g in fresh fish meat. Average contents of putrescine, cadaverine, histamine, and tyramine in tested samples were higher than those of tryptamine, 2-phenylamine, spermidine, and spermine. Four fish meal samples (10%, 4/40) and 1 FSC sample (20%, 1/5) had histamine levels >20 mg/100 g. Five histamine-producing bacterial strains isolated from fish meal samples producing 1.31–6.21 ppm of histamine in trypticase soy broth supplemented with 1.0% l-histidine (TSBH) were identified as Bacillus licheniformis (three strains), B. amyloliquefaciens (one strain), and B. subtilis (one strain).     

Effects of dietary Lactobicillus plantarum on the growth,carcass quality,and immune response of African catfish (Clarias gariepinus) challenged with Salmonella typhi

An experiment was conducted to investigate the effects of Lactobacillus plantarum on the production of African catfish (Clarias gariepinus). Five experimental diets containing 0, 10³, 10⁵, 10⁷, and 10⁹ CFU of Lactobacillus plantarum g^?1 diet (T₁–T₅ treatments respectively) were fed to African catfish (Clarias gariepinus) (9.20 ± 0.1 g initial body mass), for 70 days. Results indicated significantly lower growth performance in the fish fed diet without probiotics and in those fed diet with the least probiotic level. Treatments with probiotics significantly improved the blood profile and carcass protein but significantly reduced the carcass fat (P < 0.05); these treatments also marginally improved the carcass minerals in comparison with the treatment without probiotic (P < 0.05). Challenging the fish fed the experimental diets with Salmonella typhi showed higher immunity of fish fed the probiotic diets than those fed the nonprobiotic diet. Duncan’s multiple range test showed that the best fish performance was observed with 10³ CFU g^?1 L. plantarum for very parameter measured. However, regression analyses showed the optimum level of the bacteria to be 10^4.13–10⁵ CFU g^?1     

Quality of Filleted Atlantic Mackerel (Scomber Scombrus) During Chilled and Frozen Storage: Changes in Lipids,Vitamin D,Proteins, and Small Metabolites,including Biogenic Amines

Quality changes of vacuum-packed Atlantic mackerel (Scomber scombrus) fillets during 12 months’ frozen storage at ?27°C and 9 days’ chilled storage at +4°C were evaluated. Freezing at ?27°C preserved the long chain n-3 polyunsaturated fatty acids (LC n-3 PUFAs), both in light and dark muscle, vitamin D, and the low molecular weight metabolites (LMW) (studied by high resolution nuclear magnetic resonance spectroscopy, HR NMR). Protein oxidation took place, especially between 1 and 7 months, decreasing water holding capacity and protein extractability. During chilled storage, no lipid or protein oxidation was observed, but lipolysis increased, and several LMW metabolites relevant for sensory and nutritional quality degraded into non-favorable compounds. The content of biogenic amines was high at day 9 (e.g., 18 mg histamine/100 g), jeopardizing safety. Preservation of mackerel fillets by freezing at ?27°C is thus a better option compared to prolonged chilled storage at +4°C; the quality was well preserved for 12 months’ frozen storage.     

Eye structure and function in the giant freshwater prawn Macrobrachium rosenbergii

We used histological and electroretinogram methods to investigate eye structure and function in Macrobrachium rosenbergii. M. rosenbergii was demonstrated to have superposition compound eyes with a clear zone. Rhabdom volume dramatically increased only under dark-adapted conditions at night. The eyes were assumed to be dark adapted after about 50 min under dark conditions, and to be light adapted after about 4 min under light conditions during the day. The mean angle between receptor units was 1.75°. The slope of the visual sensitivity (V/log I) curve of M. rosenbergii was 0.40 and the K value (stimulus irradiance eliciting half of the maximum response) was 16.1 log photons cm^?2 s^?1. The mean dynamic range was 4.06, ranging from 13.91 to 17.97 log photons cm^?2 s^?1. The response latency was 19.6 ± 7.5 ms (mean ± SD). Analysis of the visual spectral sensitivity of dark-adapted specimens indicated that the sensitivity maximum was centered at a wavelength of 563 nm in the yellow–green region of the spectrum. The critical flicker fusion frequency increased as stimulus irradiance increased, up to a maximum of 44.3 Hz at almost the same stimulus irradiance of the K value.     

Effect of the UV-C Radiation on Shelf Life of Vacuum-Packed Refrigerated Pirarucu (Arapaima gigas) Fillets

The effect of Ultraviolet radiation type C (UV-C) radiation (0.100 ± 0.010 J/cm²) on shelf life of Arapaima gigas fillets stored at 4 ± 1°C for 18 days was investigated. The samples were analyzed for total aerobic mesophilic bacteria (TAMB) counts; total aerobic psychrotrophic bacteria (TAPB) counts; Enterobacteriaceae; purge loss; pH; lipid oxidation; total volatile bases (TVB-N); ammonia; biogenic amines (BAs); and L*, a*, and b* values. UV-C radiation increased (P < 0.05) the generation time of all bacterial groups evaluated, increasing the shelf life by 2 days. UV-C-treated fillets showed lower (P < 0.05) L* values, purge loss, pH, TVB-N, ammonia, and BAs levels than their control counterparts throughout the storage period. Moreover, UV-C treatment did not result (P > 0.05) in detrimental effects on lipid oxidation or a* and b* values. UV-C at 0.100 J/cm² demonstrated a good potential for use in A. gigas fillets and, therefore, it could be applied at industrial scale.     

常见水产品中生物胺的调查及分析

为研究不同水产品中生物胺的产生情况,利用高效液相色谱-柱后衍生-荧光检测技术对蓝点马鲛、黄鳍金枪鱼、银鲑、牙鲆、中国明对虾、鹰爪虾、中华绒螯蟹、杂色蛤、杂色鲍中的7种生物胺含量进行调查与分析。试验结果表明,鲜活水产品中几乎不含生物胺,或仅含少量的精胺、亚精胺;腐败水产品中检出大量的生物胺,其中蓝点马鲛、黄鳍金枪鱼、银鲑、牙鲆中的组胺含量分别为422.57、640.00、309.63、151.86mg/kg,而虾、蟹、贝类等无脊椎动物几乎不产生组胺。不同水产品在腐败过程中产生的生物胺种类、含量、比例不同。生物胺总量可作为水产品腐败变质程度的指标。     

Quality Changes in Chum Salmon (Oncorhynchus keta) Caviar (ikura) Affected by Thermal Pasteurization,Storage Time,and Packaging Material

Changes in quality parameters including pH, water activity, texture, and lipid oxidation were studied in pasteurized chum salmon (Oncorhynchus keta) ikura samples packaged using two films with different oxygen transmission rates (OTR) (40 and 62 cm³·m^?2·day^?1; F-40 and F-62), during 60 days storage at 4°C. No significant differences in pH and water activity (a_w) were observed between ikura packaged using two different films with different OTR (P > 0.05). However, compared to the first day of study, water activity decreased significantly in ikura (P < 0.05). Ikura thiobarbituric reactive substance (TBARS) in the pouches significantly increased during the storage at 4°C (P < 0.05). Both pouches showed similar trends in TBARS until day 29, while after day 29, ikura packaged in F-62 (OTR = 62 cm³·m^?2·day^?1) showed a significant increase in TBARS compared to F-40 with less OTR (P < 0.05). The texture of ikura became softer compared to the first day; however, no significant difference was observed between the ikura samples in two pouches (P < 0.05). The quality changes of ikura measured during storage indicate that packaging ikura in a lower OTR film would provide greater quality retention than one with higher OTR.