Chemical and sensory changes associated Yu-lu fermentation process – A traditional Chinese fish sauce

The production of ‘Yu-lu’, a Chinese traditional fermented fish sauce, was replicated in the laboratory with little adaption in order to study the chemical, microorganism and sensory changes associated with the process. Yu-lu was made by incubating mixtures of small anchovies and 30% salt (salt/fish, w/w) at 30 ± 5 °C for 180 days, then rising the incubation temperature to 50 °C for 7 days. Changes in total soluble nitrogen (TSN), TCA (trichloroacetic acid) soluble peptides, formaldehyde nitrogen, total acid, total volatile base nitrogen (TVB-N), trimethylamine nitrogen (TMA-N), composition of amino acid, the non-enzymatic browning index and total plate counts of Yu-lu were observed. TSN, formaldehyde nitrogen, TCA soluble peptides, total titratable acid and non-enzymatic browning index increased throughout the fermentation period. Glutamic acid, lysine, leucine and valine were prominent in Yu-lu. The results from QDA test showed that incubation after 180 days for a week can accelerate the flavor formation of Yu-lu.     

Effects of the addition of spleen of skipjack tuna (Katsuwonus pelamis) on the liquefaction and characteristics of fish sauce made from sardine (Sardinella gibbosa)

The effects of the addition of spleen of skipjack tuna (Katsuwonus pelamis), at levels of 0%, 10% and 20%, on the liquefaction and characteristics of fish sauce produced from the sardine (Sardinella gibbosa) with different salt concentrations (15%, 20% and 25%) were monitored during fermentation for 180 days. Fish sauces prepared from sardine with spleen supplementation contained greater total nitrogen, amino nitrogen, formaldehyde nitrogen and ammonia nitrogen than did those without spleen addition throughout the fermentation. The rate of liquefaction was dependent on the amount of spleen added. Reduction of salt content accelerated the hydrolysis of fish protein during fermentation. The liquefaction rate of the lower salt-treated samples was generally faster than were those treated with higher salt content. Among all treatments, sardine with 25% spleen and 15% salt added exhibited the greatest protein hydrolysis, particularly at the early stages, suggesting the combined effects of autolysis and spleen proteinase. The greater liquefaction was coincidental with the development of browning as well as the increase in redness of liquid formed. An acceptability test revealed that the samples were different in colour, aroma, taste and overall acceptance (p < 0.05). Fish sauce samples containing 20% salt, without and with 10% spleen addition had similar acceptabilities to commercial fish sauce. Therefore, the addition of spleen, as well as salt reduction, can accelerate the liquefaction of sardine for fish sauce production.     

Novel starter cultures to inhibit biogenic amines accumulation during fish sauce fermentation

Bacteria with amine oxidase activity have become a particular interest to reduce biogenic amines concentration in food products such as meat and fish sausages. However, little information is available regarding the application of these bacteria in fish sauce. Hence, our study was aimed to investigate the effect of such starter cultures in reducing biogenic amines accumulation during fish sauce fermentation. Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05 isolated from fish sauce which possess amine oxidase activity were used as starter cultures in this study. Fermentation was held for 120 days at 35 °C. The pH value increased in all samples, while salt concentration remained constant throughout fermentation. Aerobic bacteria count was significantly lower (p < 0.05) in the control than in inoculated samples as a result of starter cultures addition. However, it decreased during fermentation due to the growth inhibition by high salt concentration. Proteolytic bacterial count decreased during fermentation with no significant difference (p > 0.05) among samples. These bacteria hydrolyzed protein in anchovy to produce free amino acid precursors for amines formation by decarboxylase bacteria. The presence of biogenic amines producing bacteria in this study was considered to be indigenous from raw material or contamination during fermentation, since our cultures were negative histamine producers. Amino acid histidine, arginine, lysine and tyrosine concentration decreased at different rates during fermentation as they were converted into their respective amines. In general, biogenic amines concentration namely histamine, putrescine, cadaverine and tyramine increased throughout fermentation. However, their concentrations were markedly higher (p < 0.05) in the control (without starter cultures) as compared to the samples treated with starter cultures. Histamine concentration was reduced by 27.7% and 15.4% by Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively. Both cultures could also reduce other amines during fermentation. After 120 days of fermentation, the overall biogenic amines concentration was 15.9% and 12.5% less in samples inoculated with Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively, as compared to control samples. These findings emphasized that application of starter cultures with amines oxidase activity in fish sauce fermentation was found to be effective in reducing biogenic amines accumulation.     

低盐外加曲发酵潮汕鱼露的理化性质变化

以鳀鱼(Engraulis japonicus)为原料,在不同盐质量分数(10%,15%,20%,30%)和不同曲质量分数(0%,5%)条件下发酵30d,然后再增加盐量至30%后继续发酵180d,对整个发酵过程中的总可溶性氮(TSN)、氨基酸态氮(AA-N)、挥发性盐基氮(TVB-N)、pH值、非酶褐变指数、游离氨基酸组成、感官及色差变化进行分析。结果表明,低盐发酵及低盐外加曲发酵可加速潮汕鱼露发酵;与低盐发酵鱼露相比,低盐加曲发酵鱼露中TSN、氨基酸态氮以及主要氨基酸(谷氨酸、丙氨酸、赖氨酸)质量浓度有较大提高,其中加盐15%加曲5%工艺风味较好。     

Effect of electrodialysis pretreatment on physicochemical properties and morphology of spray-dried-fish sauce powder

Fish sauce is a popular Asian food ingredient having specific flavor that gives pleasure to the senses of taste. However, fish sauce contains high-salt content, which is nowadays not desirable by health-conscious consumers. Recently, it has been shown that electrodialysis succeeds in reducing the salt content of fish sauce without affecting its important characteristics over a specific range of residual salt content. In this study the effect of electrodialysis pretreatment prior to spray drying to produce low-salt fish sauce powder, which is a more shelf-stable and convenient product compared with liquid fish sauce, was investigated. Product recovery, morphology and selected physicochemical properties, i.e., moisture content, water activity, hygroscopicity, as well as X-ray diffraction (XRD) patterns of the fish sauce powder were determined. The ED-treated-fish sauces of various salt contents (in the range of approximately 2–25% w/w) were spray dried at inlet drying air temperatures of 120–180 °C. It was found that the feed salt concentration and inlet air temperature had significant effects on all observed variables. This is due to the fact that the feed salt concentration strongly affected water removal from the ED-treated-fish sauce during spray drying, leading to significant alterations of the ED-treated-fish sauce powder properties.     

Effects of brine concentration on shelf-life of hot-smoked tilapia (Oreochromis niloticus) stored at 4 °C

This work evaluated the effect of brine concentration on the shelf-life of hot-smoked tilapia (Oreochromis niloticus) stored at 4 °C. The fish were brined in solutions of 5%, 10%, and 15% NaCl and unsalted fish were used as controls. The fish were then smoked, cooled and stored at 4 °C. Oxidative rancidity measured by the peroxide value (PV), and thiobarbituric acid number (TBA) showed increases with the storage time and also as a result of the increasing salt content in fish muscle. Hot smoked tilapia can be stored safely under refrigerated conditions for over 35 days, and 5% brine was found to be optimal for storage.     

Optimisation of gelatine extraction from hoki (Macruronus novaezelandiae) skins and measurement of gel strength and SDS–PAGE

The extraction of hoki (Macruronus novaezelandiae) gelatine was optimised using Response Surface Methodology (RSM) and gel strength and SDS–PAGE were evaluated. The optimum conditions for extraction were 0.75 M NaCl for 9 min of pre-treatment time and hot water extraction at 49.3 °C for 60 min. Results showed that the predicted yield by RSM (17.4%) closely matched the experimental yield of 17.6%. SDS–PAGE showed that hoki gelatine contained higher molecular weight subunits (∼191 kDa) but lower gel strength (197 ± 5 g) than those from porcine (307 ± 8.4 g) or bovine (273 ± 16.1 g) gelatine determined at 7 °C. However, hoki gelatine gel strength was significantly higher than those from other cold-water fish species reported in literature, which could account for the slight differences in methodologies reported.     

Histamine contents of fermented fish products in Taiwan and isolation of histamine-forming bacteria

Twenty-seven imported fermented fish products from Southeast Asian countries and sold in the supermarkets in Taiwan, including fish sauce, fish paste and shrimp paste, were tested to determine the occurrence of histamine and histamine-forming bacteria. The levels of pH, salt content, total volatile basic nitrogen, trimethylamine, and aerobic plate count in all samples ranged from 4.8% to 6.5%, 16.2% to 45.3%, 51 to 275 mg/100 g, 5.4 to 53.9 mg/100 g and 1.0 to 4.2 log CFU/g, respectively. The average content for each of eight different biogenic amines in all samples was less than 90 ppm, except for histamine which has an average content of 394 ppm in fish sauce, 263 ppm in fish paste, and 382 ppm in shrimp paste. Most of the tested fermented fish products (92.6%) had histamine levels greater than the FDA guideline of 50 ppm, while seven of them (25.9%) contained >500 ppm of histamine. Although Bacillus coagulans and Bacillus megaterium were identified as the two histamine-producing bacteria capable of producing 13.7 and 8.1 ppm of histamine, respectively, in trypticase soy broth broth supplemented with 1.0% l-histidine, they were not determined to be the main contributors to histamine accumulation in these fermented fish products.     

The effects of packaging method (vacuum pouch vs. plastic tray) on spoilage in a cook-chill pork-based dish kept under refrigeration

The effects of two packaging methods on the spoilage of a cook-chill pork-based dish kept under refrigeration were studied. Raw pork cuts and pre-cooked tomato sauce were packed under vacuum “sous vide” in polyamide–polypropylene pouches (SV) or into translucent polypropylene trays under modified atmosphere (80% N₂ + 20% CO₂) and sealed with a top film (PT). Samples were cooked inside the pack at an oven temperature/time of 70 °C/7 h, chilled at 3 °C and stored at 2 °C for up to 90 days. Microbial (psychrotrophs, lactic-acid bacteria, Enterobacteriaceae, moulds and yeasts), physical–chemical (pH, water activity and total acidity) and sensory (colour, odour, flavour, texture and acceptance) parameters were determined. Heat penetration was faster in SV (2 °C/min) than in PT (1 °C/min) (core temperature). Both packaging methods were equally effective in protecting against microbial spoilage for 90 day at 2 °C. Minor counts were only detected for lactic-acid bacteria and anaerobic psychrotrophs in SV. No Enterobacteriaceae growth was found. Slight differences between SV and PT in pH and total acidity were observed. SV and PT had similar effects on the sensory preservation of the dishes. A gradual loss of acceptance of the cooked pork and tomato sauce was observed. Rancid flavour in PT and warmed-over-flavour in SV were noted in the final stages of storage. According to acceptance scores, the shelf-life of both SV and PT was 56 days at 2 °C. Both packaging methods can be used to manufacture sous vide meat-based dishes subsequently stored under refrigeration for catering use.     

Characterization of Ca-activated cell-bound proteinase from Virgibacillus sp. SK37 isolated from fish sauce fermentation

Cell-bound proteinase from Virgibacillus sp. SK37 isolated from the first month of fish sauce fermentation was characterized. The enzyme showed the maximum activity at 65 °C, pH 7.0 and 9.5, using azocasein as a substrate. The enzyme required at least 10 mmol/l Ca²⁺ to effectively hydrolyze casein and the extent of casein degradation increased with Ca²⁺ concentration. Ethylenediaminetetraacetic acid (EDTA) and phenylmethanesulfonyl fluoride (PMSF) largely inhibited the activity, indicating a characteristic of Ca²⁺-activated serine proteinase. Among six synthetic substrates tested, the enzyme preferably hydrolyzed Suc-Ala-Ala-Pro-Phe-AMC, indicating a subtilisin-like proteinase. Although activity towards actomyosin at 20 g/100 ml NaCl decreased to 63% compared to at 5 g/100 ml, the enzyme showed high stability at 25 g/100 ml NaCl, 30 °C. This was the first study to report biochemical characteristics of cell-bound proteinase from a moderately halophilic bacterium isolated from fish sauce.     

Study on the properties of gelatins from skin of carp (Cyprinus carpio) caught in winter and summer season

The objective of this work was to compare the physiochemical (molecular weight distribution and amino acid composition) and rheological (viscosity property, gel strength and melting point) properties of gelatins from skins of carp caught in winter to those obtained for the summer equivalents. Gelatins from winter and summer fish skins were extracted at 60, 70 and 80 °C. SDS-PAGE patterns for gelatins extracted under the same conditions showed that the degradation of gelatins from winter fish skins were more severe than that of the summer ones. The imino acid contents of the winter and summer gelatins extracted at 60 °C were very similar, showing 190 and 188 residues/1000 residues, respectively. The gelatins from summer fish presented higher melting points and gel strengths, as well as better viscosity properties than the winter equivalents (P < 0.05). The differences in the rheological properties between winter and summer gelatins may be explained by different thermostability of interstitial collagen molecules (from which gelatins were derived) in the two seasons.     

Influence of brine salting prior to pickle salting in the manufacturing of various salted-dried fish species

In Portugal, the production of salted-dried cod and similar species is a relevant fish industry. Pickle salting is the most common salting method in this industry. The objective of this work was to study the influence of brine salting before pickle salting in the characteristics of the final salted-dried product of different fish species. Three salting methods were studied: only pickle salting, brine salting at 16°Baumé followed by pickle salting and brine salting at 25°Baumé followed by pickle salting. Fish species studied were Gadus morhua, Gadus macrocephalus, Theragra chalcogramma. The results showed from 4.7% to 9.2% significant higher yields of salted-dried fish when using the lower salt content brine, but drying time increased from 32 to 45 h (35–65%), depending on size of the fish, and texture became significantly softer, what may imply that salted-dried fish using lower salt content brines may be perceived by consumers as a different product. No significant differences of color were obtained.     

Protein hydrolysates from meriga (Cirrhinus mrigala) egg and evaluation of their functional properties

Protein hydrolysates from underutilised meriga (Cirrhinus mrigala) fish egg were prepared by using commercial Alcalase and papain enzymes. The degree of hydrolysis was 62% for Alcalase and 17.1% for papain, after 90 min digestion at 50–55 and 60–65 °C, respectively. The protein content of Alcalase-produced hydrolysate was higher (85%) than that of papain hydrolysate (70%) (p < 0.05). Hydrolysis by both enzymes increased protein solubility of fish egg protein hydrolysates to above 72.4% over a wide pH range (2–12). Results showed that the hydrolysates had good fat absorption capacity (0.9 and 1.0 g/g sample), foam capacity (70% and 25%) and emulsifying capacity (4.25 and 5.98 ml/g hydrolysate), respectively for Alcalase and papain protein hydrolysates. Gel filtration chromatograms and SDS–PAGE analysis indicated the distribution of smaller peptides. These results suggested that fish egg protein hydrolysates could be useful in the food industry.     

Proteinase-producing halophilic lactic acid bacteria isolated from fish sauce fermentation and their ability to produce volatile compounds

Halophilic lactic acid bacteria were isolated from fish sauce mashes fermented at 1 to 12 months. Seven out of sixty-four isolates were selected according to their proteolytic activity and growth at 25% NaCl for characterization and investigation of volatile compound production. All selected isolates were Gram-positive cocci with pairs/tetrads and grew at 0-25% NaCl, pH 4.5-9.0. Results of 16S rRNA gene sequence analysis showed 99% homology to Tetragenococcus halophilus ATCC 33315. The restriction fragment length polymorphism (RFLP) patterns of all isolates were also similar to those of T. halophilus ATCC 33315. These isolates were, thus, identified as T. halophilus. All isolates hydrolyzed fish protein in the medium containing 25% NaCl. Intracellular aminopeptidase of 7 isolates exhibited the highest activity of 2.85-3.67 U/ml toward Ala-p-nitroanilide (Ala-pNA). T.halophilus strains MS33 and M11 showed the highest alanyl aminopeptidase activity (P < 0.05), and produced histamine in mGYP broth containing 5 and 25% NaCl in the level of 6.62-22.55 and 13.14-20.39 mg/100 ml, respectively. Predominant volatile compounds of fish broth containing 25% NaCl inoculated with T. halophilus MS33 and MRC5-5-2 were 1-propanol, 2-methylpropanal, and benzaldehyde, corresponding to major volatile compounds in fish sauce. T.halophilus appeared to play an important role in volatile compound formation during fish sauce fermentation.     

Evaluation of some physico-chemical properties of restructured trout and hake mince during cold gelation and chilled storage

Cold gelation was carried out on trout (Oncorhynchus mykiss) or on hake (Merluccius merluccius) mince with or without addition of fish oil and using microbial transglutaminase (MTGase). Products were stored at 4 °C for 6 days and lipid oxidation, protein oxidation, texture, water binding capacity, and colour were followed. Results indicated that MTGase was able to generate gels with good properties for both trout and hake. Gels prepared with trout were oxidised whilst gels prepared with hake were stable toward oxidation even in the presence of 5% fish oil. However, in the presence of oil, as an alternative for generating omega-3 enriched products, the activity of MTGase was impaired, as the gels took longer to reach maximum hardness. Furthermore, in all samples containing MTGase, protein oxidation was high.     

Chemical and biochemical changes of hybrid catfish fillet stored at 4 °C and its gel properties

Chemical and biochemical changes of aquacultured hybrid catfish fillet (Clarias macrocephalus × Clarias gariepinus) and its gel-forming ability as affected by age and sex of fish along with storage time were investigated. Fillets were stored at 4 °C for 0, 3, 6, 9, 12 and 15 days. There was no significant effect of sex and age of fish as well as storage time on fat, moisture and ash contents (P > 0.05). The total protein, water soluble protein, and salt soluble protein contents of the fillets significantly decreased with storage time (P < 0.05). On the other hand, pH, total volatile base nitrogen (TVB-N) and autolytic degradation products (ADP) increased as storage time continued (P < 0.05). Decreases in Ca²⁺-ATPase activity and gel properties were observed as storage time increased. However, there was no significant effect of either sex or age of fish on textural properties of gel (P > 0.05). Hybrid catfish fillet stored at 4 °C should be processed within 6 days.     

Chemical, microbiological and sensory changes associated with fish sauce processing

The production of sardine fermented fish sauce was replicated in the laboratory in order to study the chemical, microbiological and sensory changes associated with the process. Fish sauce were produced by incubating mixtures of sardine (Sardina pilchardus) at different concentrations of sodium chloride and glucose at 37 °C for 57 days. Changes in chemical composition (moisture, protein, fat contents), pH, total volatile basic nitrogen (TVB-N, mg N/100 g fish flesh), trimethylamine (TMA-N, mg/100 g fish flesh), thiobarbituric acid (TBA, mg malonaldehyde/kg fish flesh), water activity (aw), color measurement, total viable count, lactic acid bacteria count, yeast and moulds counts and sensory analyses were observed. The fish sauces with spices were determined lower bacteria counts than fish sauces without spices. The addition of glucose to the fish sauces caused a significant increasing in the bacterial counts. Staphylococcus aureus and yeast-mould counts were not detected during fermentation period. The use of spices in fermentation of sardine enhanced good color, aroma and taste.     

Effects of pomegranate sauce on quality of marinated anchovy during refrigerated storage

Effects of pomegranate sauce on the quality of marinated anchovy during storage at 4 °C were investigated. Anchovy were marinated with 30 g/L acetic acid and 150 g/L salt, put into glass jars, filled with either sunflower oil or pomegranate sauce and stored at 4 °C. Total volatile basic nitrogen (TVB-N) and trimethylamine (TMA-N) values increased during the storage. Higher values for free fatty acid (FFA), conjugated diens (k-dien) and para-anisidine (p-Av) were found in samples with sunflower oil than those with pomegranate sauce. Samples in pomegranate sauce showed better oxidative stability. Higher taste and flavour and lower appearance scores were found for samples in pomegranate sauce than those samples in sunflower oil. It was found that pomegranate sauce was as least effective as the traditional sunflower oil to keep quality. Pomegranate sauce also produced desirable taste and flavour but the coloration should be studied in further studies.     

发酵鳀制鱼露的理化与感官性质的变化

鳀鱼中添加30%的海盐(盐/鱼,w/w)后在(30±5)℃条件下自然发酵180d,再在50℃下保温7d。定期测定发酵液的pH值、总可溶性氮(TSN)、氨基酸态氮、TCA可溶性肽、总酸、总挥发性盐基氮(TVB-N)、三甲胺氮(TMA-N)、氨基酸组成、红色指数以及非酶褐变指数,并进行感官分析(QDA-test)。结果表明,鳀鱼露发酵过程中,TSN含量、TCA可溶性肽含量、氨基酸态氮、游离氨基酸含量、红色指数以及非酶褐变指数均随着发酵时间的延长而增加,而pH值则随着发酵时间的增加而减少。TCA可溶性肽含量在保温发酵后有明显增长,TVB-N、TMA-N含量与发酵时间不呈明显的线形相关关系。谷氨酸、赖氨酸、亮氨酸、缬氨酸和丙氨酸是鱼露中最主要的氨基酸。感官分析结果表明,鱼露发酵的后期阶段,在50℃短期保温可以起到加速风味成熟与降低不良风味的作用。