Spoilage Potential of Some Contaminant Bacteria Isolated from Indian Mackerel (Rastrelliger kanagurta)

Pseudomonas marinoglutinosa, Aeromonas hydrophila isolated from unirradiated spoiling Indian mackerel (Rastrelliger kanagurta) and Bacillus megaterium, Micrococcus colpogenes isolated from gamma-irradiated mackerel were examined for their spoilage potential in fish homogenate and fish muscle sarcoplasmic and myofibrillar protein fractions. Pseudomonas marinoglutinosa (0° -2°C), Aeromonas hydrophila (0° - 2°C) and Bacillus megaterium (26°C) proliferated in the fish homogenate and sacroplasmic protein fraction which was associated with losses in sensory attributes and increase in non-protein-nitrogen (NPN) compounds, total volatile acids (TVA) and total volatile basic nitrogen (TVBN) compounds. On the other hand, the organisms exhibited diminished growth in MF fraction which was also accompanied by less changes in sensory and chemical quality parameters. Growth of M. colpogenes was not accompanied by putrefaction of the fish media as assessed by sensory and chemical criteria, suggesting contribution of varying spoilage potentials of the bacterial isolates in the overall spoilage of the fish.     

ROLE OF BACTERIAL PROTEOLYSIS IN THE SPOILAGE OF IRRADIATED FLESH FOODS

Microorganisms (Aeromonas hydrophila, Bacillus megaterium, Pseudomonas marinoglutinosa and Salmonella typhimurium) proliferated equally well when incubated in unirradiated and irradiated fish myofibrillar protein fractions. However, formation of Total Volatile Bases was found to be 50–60% less in the irradiated myofibrillar protein fraction. Proteolysis measured in terms of tyrosine release also registered 50% less in irradiated flesh foods as compared to unirradiated samples under identical conditions. Bacterial protease levels were also evaluated after growth in both control and irradiated fish proteins using hemoglobin as substrate which revealed that there is considerable reduction in protease secretion when the bacterium proliferated in irradiated fish proteins. These results suggest that appreciable delay in the spoilage of irradiated flesh foods could be attributed to low bacterial proteolysis.     

ADIATION SENSITIVITY AND BIOCHEMICAL CHARACTERISTICS OF MICROFLORA OF BOMBAY DUCK (Harpodon nehereus)

SUMMARY– The microbial flora of unirradiated and irradiated (0.5 Mrad) Bombay duck (Harpodon nehereus) was differentiated into organisms as spoilers and non-spoilers based on ability to liquify gelatin; ferment glucose; and produce indole, H₂S and urease. In spoiling unirradiated fish, there was a predominance of Vibrio, Aeromonas and Proteus spp while Micrococci and Achromobacter spp were the major surviving groups in irradiated Bombay duck stored for 15 days at 10°C. The predominant spoilers, Proteus vulgaris and Aeromonas hydrophila, produced large amounts of TMA and TVBN and were radiation sensitive as indicated by D₁₀ values of 8.6 and 5.4 Krad respectively. Micrococcus luteus was relatively biochemically inert and radiation resistant, the D₁₀ value being 88 Krad.     

Sensory,microbiological, and chemical spoilage of cultured common sea bass (<Emphasis Type="Italic"> Dicentrarchus labrax</Emphasis>) stored in ice: a seasonal differentiation

Common sea bass ( Dicentrarchus labrax) of a Greek cage-culture origin, sampled in December and July, were stored in ice, and their sensory, microbiological and chemical spoilage patterns were studied as well as their seasonal differentiation. The sensory storage life was determined at 15 days of ice storage. The microbial population in the muscle reached levels of 10⁵ at the acceptability limit. The ATP breakdown pattern showed a quick depletion of inosine-monophosphate (IMP) in the first 8–9 days of ice storage and slower depletion thereafter. Inosine (INO) and hypoxanthine (Hx) were formed at significant quantities. TVBN and TBA, as indicators of protein breakdown and lipid oxidation, respectively, showed low and delayed increase. A seasonal effect was observed with summer fish showing higher rates of K-value increase during early spoilage, while winter fish showed higher K-values, microbial populations and TVBN at late spoilage stages.     

RADIATION PASTEURIZATION OF FRESH AND BLANCHED TROPICAL SHRIMPS

SUMMARY— The storage life of fresh, peeled and deveined tropical shrimps at 10—12°C is extended to 10—14 and 18—21 days with 0.15 and 0.25 Mrad, respectively, as against spoilage of unirradiated samples within 4 days. Blanching treatment (steaming for 4 min) with or without brine (3.0% NaCl) extends the storage life up to 30 days at lo—12°C. the former retaining the shell-fish flavor. However, the blanched products develop musty odor and slimy texture during storage; subjected to irradiation (0.15 Mrad) these are devoid of undesirable attributes and are acceptable for 60 and 130 days, respectively, at 10—1°C and 2—4°C. Bacterial counts exceeding 10⁸/g and high levels of TMAN and TVBN concur with spoilage of unirradiated shrimps. In contrast, low levels of TMAN and TVBN, despite high TBC, corroborate with high acceptability of radiation processed shrimps. In blanched as well as in blanched and irradiated shrimps TBC, TMAN and TVBN are low. Samples subjected to blanching only have low acceptability as against the improvement in quality by irradiation. These alterations in storage property have been discussed in the light of shifts in microflora.     

Modeling total volatile basic nitrogen production as a dose function in gamma irradiated refrigerated squid rings

The behavior of a chemical spoilage index for marine products (total volatile basic nitrogen – TVBN) was modeled as a function of the irradiation dose in Illex argentinus rings stored in refrigeration. The effect of gamma irradiation at 0, 1.8, 3.3 and 5.8 kGy on TVBN was analyzed in vacuum-packed squid rings during storage at 4 ± 1 °C. The modified Gompertz model satisfactorily described TVBN behavior for each irradiation dose (R² > 0.980; RMSE < 5.7). Gompertz model parameters (μ, A, L) were modeled with dose-dependent second order polynomials, for doses ranging between 0 and 5.8 kGy, in order to develop a complete model that is useful for predicting TVBN as a function of the dose in squid rings. Model validation was carried out using independent data of TVBN in squid rings irradiated at 4.8 kGy. The developed model can be used to predict TVBN production in gamma irradiated squid meat.     

鱼类腐败菌腐败能力测定方法

通过分析比较接种腐败菌的大黄鱼无菌鱼块和灭菌鱼汁,在贮藏中感官、挥发性盐基氮(total volatile base nitrogen,TVBN)、三甲胺(trimethylamine,TMA)和腐败菌的变化,以及腐败菌生长动力学参数和腐败代谢产物的产量因子(YTVBN/CFU 和YTMA/CFU),探讨无菌鱼块和灭菌鱼汁两种腐败能力的测定方法。结果表明：接种腐败希瓦氏菌的无菌鱼块和灭菌鱼汁的货架期分别为162h 和132h,此时的TVBN 含量分别为32.16mg/100g 和29.64mg/100mL,TMA含量为9.31mg/100g 和0.99mg/100mL,腐败希瓦氏菌菌数为8.67 lg(CFU/g)和8.56 lg(CFU/mL),产量因子YTVBN/CFU为2.58 × 10-10mg TVBN/CFU 和1.98 × 10-10mL TVBN/CFU,产量因子YTMA/CFU 为2.12 × 10-10mg TMA/CFU 和1.27× 10-11mL TMA/CFU。TMA 作为接种鱼汁的理化指标是不可靠的,而TVBN 可以作为接种鱼汁的理化指标。接种腐败希瓦氏菌的无菌鱼块和灭菌鱼汁产量因子YTVBN/CFU 的相对误差为23.64%,因此灭菌鱼汁作为腐败菌腐败能力测定方法具有一定的可靠性。     

Nucleotide degradation products of gamma‐irradiated sea bream (Sparus aurata) stored in ice

Nucleotide degradation products of irradiated sea bream stored up to 19 days in ice were investigated. Irradiation had significant effect on the nucleotide concentrations in sea bream muscle (P < 0.05). The results showed that the highest value of inosine monophosphate (IMP) was observed in irradiated sea bream at 5 kGy, followed by at 2.5 kGy. Initial inosine (INO) concentration in irradiated sea bream at 5 kGy was 4.26 μmoles g^?1, which reached maximum value of 8.83 μmoles g^?1 when fish completely spoiled (19 days). When the fish reached the limit of acceptability, the mean values of K, Ki, H and G were 86.8%, 90.3%, 59.8% and 213.9% for unirradiated sea bream, 85.2%, 87.8%, 56.8% and 197.8% for irradiated sea bream at 2.5 kGy and 88.4%, 90.9%, 57.8%, 211.5% for irradiated sea bream at 5 kGy, respectively. The results of this study indicated that nucleotide degradation was more rapid in unirradiated sea bream than those irradiated. K, Ki, H and G value in irradiated fish can be used as a freshness index because there is a good linear relationship between values and storage time of fish.     

Microbiological Method to Identify Irradiated Meat

Beef, chicken, mutton and pork exposed to -γ-radiation doses of 0–5 kGy were inoculated with Aeromonas hydrophila initially, after 7 days and 15 days of storage at 3°C and -11°C. After 18 hr incubation at 30°C TVBN values of nonirradiated samples were found to be 210 mg% and TVA value were 205 while the corresponding irradiated samples showed 30–80 mg% and 40–80. A similar pattern was observed in stored samples. A rapid method was developed by incubating chicken meat at 37°C for 6–7 hr with bacteria followed by estimation of volatile acids (TVA) and bases (TVBN). A 40–50% reduction in TVA and TVBN in irradiated samples was observed suggesting its possible application for detecting irradiated meat samples.     

Sensory, microbiological, physical and chemical properties of cuttlefish (Sepia officinalis) and broadtail shortfin squid (Illex coindetii) stored in ice

The objective of this study was to characterize whole raw cuttlefish (Sepia officinalis) and shortfin squid (Illex coindetii) during storage in ice through sensory, microbiological, chemical and physical analyses. The recently developed Quality Index Method (QIM) tables for these species were used for sensory analysis. Shelf-life of whole cuttlefish and shortfin squid were estimated as around 10 and 9 days, respectively, according to QIM, which is shorter than for most species, especially fish. The numbers of microorganisms found in cuttlefish and shortfin squid surfaces until rejection were lower than in fish, which suggest predominance of enzymatic (autolytic) degradation. H₂S-producing bacteria constituted a significant proportion of the spoilage flora. Physical analysis performed using the RT-Freshmeter and the Torrymeter showed these instruments can be applied to cuttlefish and shortfin squid, providing useful complementary information on the rates of change of electrical properties. Free tryptophan and VBN contents significantly changed during the first storage week indicating that low levels of these compounds could be used as indicators of fresh and high quality cuttlefish and broadtail shortfin squid. Urea can be useful as spoilage indicator for cuttlefish, as well as agmatine for broadtail shortfin squid. An overall look into data obtained shows, as expected, that spoilage of these cephalopod species can be considered a different phenomena when compared to fish and is not yet completely clarified.     

Gamma radiation effect on quality changes in vacuum‐packed squid (Illex argentinus) mantle rings during refrigerated (4–5 °C) storage

The effect of gamma radiation (0, 1.8, 3.3 and 5.8 kGy) on microbiological, chemical and colour characteristics of vacuum‐packed squid (Illex argentinus) mantle rings was studied. Total viable counts; psychrotrophic bacteria counts, Escherichia Coli, Staphylococcus aureus and Clostridium perfringens; total volatile basic nitrogen (TVBN) and colour difference were analysed during 29 days of storage at 4–5 °C. Higher doses of gamma radiation significantly reduced Total Viable, phychrotrophic counts and TVBN production (P < 0.05) in a dose‐dependent way, delaying squid spoilage. Colour difference of non‐irradiated samples with respect to first day significantly increased while it was constant in radiated samples during 22 days (P < 0.05). Independently from the dose, radiation avoided colour changes of squid rings. Gamma irradiation was effective in delaying deterioration reactions, improving microbiological, chemical and colour quality of vacuum‐packed squid rings stored at 4–5 °C.     

Identification of Microbial Isolates from Vacuum-Packaged Ground Pork Irradiated at 1 kGy

Bacterial cultures from irradiated (1 kGy) and nonirradiated, vacuum-packaged ground pork held at 5°C were isolated and characterized over a 12-day storage period. The initial flora of the meat was composed mostly of Pseudomonas sp. and Enterobacter sp. Although the microflora of nonirradiated samples gradually shifted from Gram-negative to Gram-positive microorganisms, 76% of the isolates were characterized as Gram-negative at the onset of spoilage (9 days at 5°C). In contrast, the irradiated ground pork microflora was mainly Gram-positive (66%) shortly after irradiation and increased to 97% after 9 days at 5°C. A total of 720 isolates were identified to genus.     

Effects of a bacteriocin-like substance produced by Leuconostoc mesenteroides subsp. cremoris on spoilage strain Lactobacillus fructivorans and various pathogens

Certain lactic acid bacteria present in the natural flora of the wine fermentation medium perform the malolactic fermentation process and other biochemical activities which lead to the formation of the outstanding aroma and bouquet of wine. However, because of the spoilage phenomena caused also by certain lactic cultures, off‐odours formed via metabolic side reactions cause severe economic losses in the wine industry. Several control mechanisms were introduced in order to eliminate these spoilage cultures, but none of these has been implemented as an efficient and healthy process. A partially purified bacteriocin‐like inhibitory substance (BLIS) with a molecular weight of 32 000 Da, obtained from a Leuconostoc mesenteroides subsp. cremoris isolate belonging to the native flora of wines of the Cappadocia region, was studied for its inhibitory activity on the spoilage strain, Lactobacillus fructivorans, isolated from the same wine flora and several other pathogens, and was found to be efficient in eliminating both the spoilage strain and the pathogens.     

The bacteriology of fresh and spoiling Lake Victorian Nile perch (Lates niloticus)

A total of 177 bacterial cultures isolated from Lake Victorian Nile Perch (Lates niloticus) were investigated. The flora on newly caught Nile perch consisted of organisms belonging to the genera Moraxella, Alcaligenes, Acinetobacter, Pseudomonas, Aeromonas, Micrococcus and other Gram-positive organisms. 39% were identified as Gram-positive species and 61% were negative in the Gram-reaction. Three cultures out of 53 investigated caused weak rotten off-odours in sterile fish broth and one culture, an Aeromonas spp. produced strong rotten, fishy, hydrogen sulphide off-odours. From Nile perch spoiled at ambient temperature, 15 of the 42 strains isolated caused rotten, fishy, hydrogen sulphide off-odours. These specific spoilage bacteria were all identified as Aeromonas and all reduced trimethylamine oxide to trimethylamine and produced hydrogen sulphide. From spoiled iced Nile perch, 74 out of 82 (90%) of the bacteria isolated were identified as Pseudomonas. A small proportion of these (13 out of 74) produced off-odours in sterile fish broth resembling the spoiling fish. These specific spoilers could not be separated from the non-spoilers based on biochemical activities used in classical taxonomy. While the Pseudomonas spp. isolated did not produce trimethylamine or H₂S, a few of the remaining isolates (two Shewanella putrefaciens and five Aeromonas spp.) did produce these compounds. The role of Shewanella putrefaciens in the iced spoilage of Nile perch was, however, insignificant, since they only very late in the storage reached numbers where their spoilage could be detected.     

冷藏鲤鱼和罗非鱼优势腐败菌腐败能力分析

通过分析接种腐败菌的鲤鱼和罗非鱼无菌鱼块贮藏中感官、腐败代谢产物和腐败菌的变化,以腐败菌的生长动力学参数和腐败代谢产物的产量因子(YTVBN/CFU)为指标,探讨冷藏鲤鱼和罗非鱼优势腐败菌假单胞菌和腐败希瓦氏菌的腐败能力。结果表明:接种腐败希瓦氏菌和恶臭假单胞菌的鲤鱼无菌鱼块的货架期分别为132h和162h,此时的TVBN值为27.12mg/100g和22.51mg/100g,腐败希瓦氏菌和恶臭假单胞菌菌数为8.96 lg(CFU/g)和9.07 lg(CFU/g),产量因子YTVBN/CFU为9.28×10-9mg TVBN/CFU和1.81×10-8mg TVBN/CFU。接种荧光假单胞菌和腐败希瓦氏菌的罗非鱼无菌鱼块的货架期分别为132h和144h,此时的TVBN值为23.46mg/100g和24.30mg/100g,荧光假单胞菌和腐败希瓦氏菌菌数为8.83 lg(CFU/g)和9.12 lg(CFU/g),产量因子YTVBN/CFU为1.67×10-8mg TVBN/CFU和9.10×10-9mg TVBN/CFU。结合两种养殖鱼冷藏过程中的菌相变化和腐败菌在腐败过程中的作用,初步得出冷藏罗非鱼和鲤鱼的特定腐败菌是假单胞菌,两种腐败菌都具有较强的腐败能力。     

Identification and stability study of irradiated chicken,pork, beef,lamb, fish and mollusk shells by electron paramagnetic resonance (EPR) spectroscopy

The application of electron paramagnetic resonance (EPR) spectroscopy to the detection of irradiation treatment of bones of chicken, pork, beef, lamb, fish and mollusk shells was described. Induced radicals in irradiated samples gave distinguishable EPR patterns for differentiation from the unirradiated samples. The EPR signals were found to be relatively stable in mammalian bones and mollusk shells; but significant loss was observed in fish samples. Dose–response relationships of the samples were found to be linear at 0–5 kGy range (r ²>0.95). Using calibration curve method, the irradiation status of all the 64 blind samples (14 unirradiated and 50 irradiated) was correctly identified. The estimation of irradiation doses was satisfactory with mean values ranging from 86 to 116% of those of the nominal values, and precision (%RSD) ranged from 15 to 29%. The validated analytical procedure was applied in the surveillance monitoring of about 400 food samples during the period of 2001–2004.     

Influence of storage temperature on the quality of salted mackerel (Rastrelliger kangurta) and pink perch (Nemipterus japonicus)

Dry-salted mackerel and pink perch were stored at two temperatures: ambient (26·8 ± 3·3°C) and 2·5 ± 1°C. Changes in moisture content, salt content, water activity (a_w), peroxide value (PV), free fatty acid content (FFA), total volatile base nitrogen (TVBN) content, halophilic bacterial count and sensory scores for overall acceptability were studied. Loss of moisture and absorption of salt were considerably higher in the products stored at ambient temperature. The decrease in a_w was more pronounced at ambient temperature than at the lower temperature. Although the chemical indices of freshness (PV, FFA and TVBN) and the halophilic counts showed increasing trends, they were considerably lower in the products stored at the lower temperature. Sensory evaluation for overall acceptability indicated that storage at the lower temperature could considerably extend the shelf-life of salted fish.     

Real-Time Monitoring of Shrimp Spoilage Using On-Package Sticker Sensor Based on Natural Dye of Curcumin

The development of a curcumin-based sensor for the detection of volatile amines (specifically known as total volatile basic nitrogen, TVBN) is described. Curcumin [(1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5dione] is the major yellow pigment extracted from turmeric, a commonly used spice, derived from the rhizome of the plant Curcuma longa. Curcumin was immobilized onto bacterial cellulose membrane via the absorption method. Thus, the sensing materials are edible and suitable for food applications. The curcumin/bacterial cellulose membrane as the TVBN sensor worked based on pH increase as the basic spoilage volatile amines produced gradually in the package headspace, and subsequently, the color of the sensor will change from yellow to orange, then to reddish orange for spoilage indication, which is easily visible to the naked eye. The curcumin membrane is a highly sensitive material toward acid–base reactions. Color changes, as a result of its interactions with increasing pH (as a result of increasing TVBN), were monitored directly with visual inspection and the color quantitatively measured with color analysis via Photoshop software. Furthermore, the membrane response was found to correlate with bacterial growth patterns in shrimp samples. Finally, the curcumin/bacterial cellulose membrane was successfully used as a sticker sensor for real-time monitoring of shrimp spoilage in ambient and chiller conditions.     

Shelf‐life extension of vacuum‐packaged sea bream (Sparus aurata) fillets by combined γ‐irradiation and refrigeration: microbiological,chemical and sensory changes

The combined effect of γ‐irradiation and refrigeration on the shelf‐life of vacuum‐packaged sea bream (Sparus aurata) fillets was studied by monitoring the microbiological, chemical and sensory changes of non‐irradiated and irradiated fish samples using low‐dose irradiation doses of 1 and 3 kGy. Fish species such as sea bream and sea bass are very popular in the Mediterranean countries due to their high quality characteristics, and their preservation is a constant challenge given their extreme perishability. Irradiation (3 kGy) dramatically reduced populations of bacteria, namely, total viable counts (3 vs 7 log cfu g^?1) for the non‐irradiated samples, Pseudomonas spp (<2 vs 7.6 log cfu g^?1), H₂S‐producing bacteria typical of Shewanella putrefaciens (<2 vs 5.9 log cfu g^?1), Enterobacteriaceae (<2 vs 6.0 log cfu g^?1) and lactic acid bacteria (<2 vs 3.5 log cfu g^?1) after 10 days of storage. The effect was more pronounced at the higher dose (3 kGy). Lactic acid bacteria, Enterobacteriaceae and H₂S‐producing bacteria typical of Shewanella putrefaciens showed higher sensitivity to γ‐radiation than did the rest of the microbial species. Of the chemical indicators of spoilage, Trimethylamine (TMA) values of non‐irradiated sea bream increased very slowly, whereas for irradiated samples significantly lower values were obtained reaching a final value of 7.9 and 6.3 mg N per 100 g muscle at 1 and 3 kGy respectively (day 42). Total volatile basic nitrogen (TVB‐N) values increased slowly attaining a value of 67.3 mg N per 100 g for non‐irradiated sea bream during refrigerated storage, whereas for irradiated fish, lower values of 52.8 and 43.1 mg N per 100 g muscle were recorded (day 42). Thiobarbituric acid (TBA) values for irradiated sea bream samples were higher than respective non‐irradiated fish and increased slowly until day 21 of storage, reaching final values of 1.1 (non‐irradiated), 2.0 (1 kGy) and 2.2 mg malonaldehyde kg^?1 muscle (3 kGy), respectively (day 42). Sensory evaluation showed a good correlation with bacterial populations. On the basis of overall acceptability scores (sensory evaluation) a shelf‐life of 28 days (3 kGy) was obtained for vacuum‐packaged sea bream, compared with a shelf‐life of 9–10 days for the non‐irradiated sample. Copyright © 2004 Society of Chemical Industry     

Biogenic and volatile amine-related qualities of three popular fish species sold at Kuwait fish markets

Three fish species commonly consumed in Kuwait were studied for their total volatile base nitrogen (TVBN) and biogenic amine composition during normal commercial activities at the fish markets. Samples of the fish species Zubaidi or silver pomfret (Pampus argenteus), Hammour or orange–spotted grouper (Epinephelus coioides), and Negrule or grunt (Pomadasys kaakan) were analyzed using steam distillation and high performance liquid chromatography (HPLC) procedures. Fish samples collected in the morning had lower levels of TVBN, ranging from 25.9 mg/100 g to 30.2 mg/100 g compared with the range of 31.2 mg/100 g to 40.9 mg/100 g for samples collected in the evening. Among all three fish species, Zubaidi (Pampus argenteus) showed the lowest TVBN content while Negrule (Pomadasys kaakan) had the highest. Biogenic amine levels were also low in the fish species. The data suggest that fish handling practices at the fish markets in Kuwait do not result in significant abuse of fish temperatures, and that the amounts of volatile and biogenic amines produced during normal activities did not rise to levels indicating fish decomposition.