Identification of pepsinogens and pepsins from the stomach of European eel (Anguilla anguilla)

Three pepsinogens (PG-I, PG-II and PG-III) were purified to homogeneity from the stomach of freshwater fish European eel (Anguilla anguilla) by ammonium sulphate fractionation, column chromatography on anion exchange and gel-filtration. Their molecular masses were determined as 36, 37 and 36 kDa, respectively, by SDS–PAGE, which were in agreement with the results obtained by Sephacryl S-200 and Superdex 75 gel-filtration. PG-I, PG-II and PG-III converted into corresponding pepsins with molecular masses of approximately 30 kDa at pH 2.0 and showed maximal activity at pH 3.5, 2.5 and 2.5. Optimal temperatures of these pepsins were 40, 40 and 35 °C, using bovine haemoglobin as substrate. Western blot analysis revealed that anti-sea bream PG-I and PG-II polyclonal antibodies cross-reacted with all three PGs of European eel. On the other hand, anti-sea bream PG-III and PG-IV antibodies cross-reacted with PG-II and PG-III of European eel, while no cross-reaction with PG-I was detected. The kinetic constants of K_m and k_cat of pepsins (P-I, P-II and P-III) for haemoglobin were calculated as 8.8 × 10⁻⁵ M, 23.7 s⁻¹; 9.2 × 10⁻⁵ M, 19.4 s⁻¹ and 7.0 × 10⁻⁵ M, 34.4 s⁻¹, respectively.     

Nucleotide degradation and biogenic amine formation of wild white grouper (Epinephelus aeneus) stored in ice and at chill temperature (4 °C)

Sensory (cooked and uncooked), chemical (proximate composition, TVB-N, nucleotide degradation products and biogenic amines) and microbiological quality (TVC and total coliform) changes were investigated during storage of ungutted white grouper kept in ice and at chill temperature (4 °C). According to the sensory assessment, the shelf life of white grouper was 16 days in ice and 4 days for fish stored at chill temperature. TVB-N values increased with storage time. Amines found in white grouper stored in ice were TMA, putrescine, cadaverine, 2-phenylethylamine, dopamine, agmatine, tryptamine and serotonin. Histamine, spermine, spermidine were never detected with either storage condition. The acceptability limit in terms of microbial count was exceeded at 8 days in ice and at 4 days for fish stored at chill temperature. Total coliform count was 2.8 log₁₀ cfu/ml at 1 day and reached 10⁵ cfu/ml for both storage conditions.     

The effect of thawing methods on the quality of eels (Anguilla anguilla)

Physical (colour), chemical (pH, total volatile basic nitrogen (TVB-N), thiobarbituric acid values (TBA)) and microbiological (total aerobic mesophylic bacteria, salmonella, coliform, yeast and mould counts) analyses were carried out on thawed European eel (Anguilla anguilla). Different thawing treatments were used (in a refrigerator, in water, in air at ambient temperature and in a microwave oven). The results obtained were compared statistically with those of fresh fish. pH, TBA and a^∗ values of thawed samples usually decreased significantly (P < 0.05) when compared to the fresh control. Salmonella was not detected in any of the samples. Coliform and mould counts of fresh control and thawed samples were <1 CFU/g. Total aerobic mesophylic bacteria count of all thawed fish decreased. However, the yeast count of the refrigerator–thawed samples increased. The lowest total aerobic mesophylic bacteria and yeast counts were determined in water–thawed samples. Water thawing is therefore suitable for frozen eel.     

Lipid and fatty acid compositions differentiate between wild and cultured Japanese eel (Anguilla japonica)

The lipid profile and fatty acid composition of muscle, liver, and plasma lipoprotein were examined in wild and cultured Japanese eel (Anguilla japonica). Although, the muscle lipid levels of wild eels (11.6%) were less than those of cultured eels (13.1%), both eels were classified as typical fatty fish. Compared with the liver lipid composition of cultured eels, triacylglycerol (TG) level of the liver decreased in wild eels, whilst phosphatidylcholine and phosphatidylethanolamine levels of the liver increased in wild eels, reflecting the difference of liver lipid levels in both eels. Wild eels contained more cholesteryl ester (CE) and less TG, phospholipid, and free cholesterol in their plasma than cultured eels. The ratio of TG to CE decreased, whilst that of CE to total cholesterol increased in the plasma of wild eels. Different fatty acid compositions were found between wild and cultured eels. Compared with the fatty acid compositions of cultured eels, wild eels contained high percentages of 18:2, 18:3, and 20:4 and low percentages of 22:6, 20:1, and 20:5 in their muscle, liver, and plasma lipoprotein. The lipid profile and fatty acid composition seemed to be useful criteria for distinguishing between wild and cultured eels.     

The biogenic amines and bacterial changes of farmed rainbow trout (Oncorhynchus mykiss) stored in ice

The biogenic amine content and related bacterial changes (Pseudomonas spp., psychrotrophic and mesophilic counts) in whole farmed rainbow trout (Oncorhynchus mykiss) were monitored during ice storage for 18 days (at 0, 3, 6, 9, 12, 15 and 18 days). Levels of putrescine, cadaverine and histamine, and bacterial loads, increased (P < 0.05) during storage, but tyramine was not detected in any of the tested samples. Concentration of putrescine ranged from 0.4 initially to 8.97 μg/g, and psychrotrophic microorganisms were dominant. The best linear regressions (correlations) were for putrescine and Pseudomonas spp. and psychrotrophs (r = 0.98), and for cadaverine with Pseudomonas spp. (r = 0.82). Putrescine content was a good quality marker. Histamine was detected only at later stages of storage and was therefore less suitable than the other biogenic amines as freshness indicator.     

鱼肉蛋白的热变性研究进展

为探究盐浓度对欧洲鳗肌原纤维蛋白热诱导凝胶形成性能的影响,测定了蛋白的浊度、表面疏水性和活性巯基的变化,考察了0.1、0.3、0.5 mol/L的NaCl浓度下的肌原纤维蛋白热诱导凝胶的性质。结果表明,欧洲鳗肌原纤维蛋白的浊度、表面疏水性和活性巯基分别在30、35和40 ℃开始增加。随着NaCl浓度的增加,肌原纤维蛋白的热吸收峰温度降低,而储能模量和损耗模量增加。0.5 mol/L NaCl溶解的欧洲鳗肌原纤维蛋白热诱导凝胶破断强度为98.81 g,高于低浓度NaCl溶解的蛋白热诱导凝胶。电泳结果表明,增加NaCl浓度会促进肌球蛋白重链和肌动蛋白的相互作用。根据傅里叶变换红外光谱和扫描电镜结果,发现0.5 mol/L NaCl溶解的肌原纤维蛋白热诱导凝胶具有最高的酰胺II/酰胺I强度比和最致密的网络结构。研究结果表明欧洲鳗肌原纤维蛋白在35 ℃时开始变性,提高NaCl浓度可以增强热诱导蛋白凝胶的强度和网络结构致密度,将为利用盐浓度和温度调控鳗鱼加热调理产品提供理论指导。     

Freshness assessment of ray fish stored in ice by biochemical,chemical and physical methods

Ray fish were caught, filleted, and stored in ice. Fillets were analysed for 18 days to determine the chemical, biochemical and physical changes and their relation to the muscle eating quality. Trimethylamine (TMA-N), total volatile bases (TVB-N), ATP content and breakdown products, K value, pH, texture, water-holding capacity (WHC) and colour changes were monitored. At the beginning of the study, the ray fish muscle showed a low concentration of ATP and a high value of inosine 5′-monophosphate (IMP). Regarding to the signs of freshness and deterioration, K value presented an exponential increase (r2 = 0.95) with an initial value of 4.7% and a final value of 47.5%. Furthermore, the TBV-N and TMA-N significantly increased (P < 0.05) during the storage in ice. As for the physical analysis whereas the texture changed (P < 0.05); pH and the WHC were not affected (P < 0.05). The overall results of this study indicated that the edible quality of ray fish muscle was maintained during at least 15 days of ice storage.     

Sensory,microbiological and chemical assessment of the freshness of red mullet (Mullus barbatus) and goldband goatfish (Upeneus moluccensis) during storage in ice

The shelf life of red mullet and goldband goatfish during ice storage were studied in terms of sensory, microbiological and chemical changes. The sensory acceptability limit was 8 days for goldband goatfish (Upeneus moluccensis) and 11 days for red mullet (Mullus barbatus) stored in ice. The TVC level was correlated with sensory assessment. The TVC exceeded 7 log cfu g⁻¹ after 8 days for goldband goatfish, and 11 days for red mullet. At the end of storage period, pH, TVB-N, TBA, FFA and PV for red mullet were 7.84, 47.19 mg/100 g, 0.69 mg MA kg⁻¹, 1.17% oleic acid and 1.58 meq O₂/kg and for goldband goatfish they were 7.53, 43.97 mg/100 g, 0.74 mg MA kg⁻¹, 1.62% oleic acid and 1.68 meq O₂/kg, respectively. In red mullet, agmatine, serotonin, histamine and dopamine became the dominant amines, reaching 7.30, 5.97, 2.52 and 2.31 mg/100 g, respectively. Also the dominant amines for goldband goatfish were 4.37, 3.88, 3.38 and 2.00 mg/100 g for histamine, agmatine, dopamine and putrescine, respectively.     

Effect of reduced oxygen atmosphere and sodium acetate treatment on the microbial quality changes of seer fish (Scomberomorus commerson) steaks stored in ice

The effect of reduced oxygen atmosphere and sodium acetate treatment on the microbial quality of seer fish (Scomberomorus commerson) steaks was determined during chilled storage (1–2 °C). The O₂ absorber reduced the oxygen content in the pack to less than 0.01% corresponding to 99.96% reduction within 24 h. The use of O₂ absorber with sodium acetate dip treatment (2% w/v) extended the sensory shelf life up to 25 days compared to only 12 days for control air packs and 20 days for untreated samples with O₂ absorber. A prominent lag phase was observed for many bacterium studied, particularly for the sodium acetate treated samples with O₂ absorber. On the day of sensory rejection, both the total mesophilic and psychrotrophic counts reached 7.7–8.1 and 7.1–7.9 log cfu/g, respectively. The sodium acetate treatment and reduced O₂ atmosphere affected the type of major spoilers. In air packed samples, H₂S-producers predominated followed by Brochothrix thermosphacta, Pseudomonas spp., where as in the untreated samples with O₂ absorber, H₂S-producers predominated the microbial flora followed by Lactobacillus spp. For treated samples with O₂ absorber, B. thermosphacta formed the major micro-flora followed by Lactobacillus spp. The use of O₂ absorber inhibited the growth of Pseudomonas spp., and total Enterobacteriaceae.     

The effects of gamma-irradiation on the nucleotide degradation compounds in sea bass (Dicentrarchus labrax) stored in ice

The effects of two irradiation doses (2.5 and 5 kGy) on nucleotide breakdown compounds in sea bass stored in ice were investigated. Nucleotide degradation was slower in sea bass irradiated at 2.5 and 5 kGy than untreated samples. Irradiated samples had lower hypoxanthine and inosine content than the control group. Significant differences (p < 0.05) in K and related values were found between control groups and irradiated samples (2.5 and 5 kGy). H and G values showed a good correlation with storage time (r² ? 0.98) while linear regressions of K and Ki ranged from 0.95 and 0.93 to 0.98 and 0.97, respectively. A dose of 5 kGy seemed to be more effective than that of 2.5 kGy to reduce nucleotide breakdown in sea bass. The best linear correlation was obtained from G and H values; thus, they might be used as freshness indicators for non-irradiated and irradiated sea bass.     

Effects of food additives on biogenic amine formation in Myeolchi-jeot, a salted and fermented anchovy (Engraulis japonicus)

This study was carried out to reduce biogenic amine contents in Myeolchi-jeot, a salted and fermented anchovy (Engraulis japonicus). The effects of various food additives on biogenic amine formation were determined by HPLC. The greatest inhibitory effect on biogenic amine production was observed in the culture treated with glycine. In the culture, the contents of putrescine, cadaverine, histamine, tyramine and spermidine were reduced by 32.6%, 78.4%, 93.2%, 100.0% and 100.0%, respectively, compared to control. The other additives tested had less effect in inhibiting biogenic amine production. Out of food additives tested, glycine was finally applied to the ripening of Myeolchi-jeotin situ, and then overall production of biogenic amines was reduced by up to 63.0% and 73.4%, compared to controls prepared with no and 20% NaCl, respectively. Therefore, it is expected that the findings of this study enhance the safety of not only Myeolchi-jeot but other salted and/or fermented seafood.     

Effect on lamb meat quality of including thyme (Thymus zygis ssp. gracilis) leaves in ewes’ diet

The aim of this study was to investigate the effect of including thyme leaves (TL) in the diet of pregnant sheep on the sensorial characteristics, bacterial spoilage and oxidative stability of lamb meat stored in modified atmosphere (70% O₂:30% CO₂). For this, thirty-six sheep were randomly assigned to three groups: control (basal diet), T₁ (3.7% thyme leaves), T₂ (7.5% thyme leaves). Meat spoilage (TV, PSY, MY, ENT, and LA), TBARS, CIELAB coordinates, metmyoglobin and the sensory characteristics of fresh lamb meat were analyzed on days 0, 7, 14 and 21. The presence of antioxidant compounds in the diet containing TL delayed (P < 0.05) colour deterioration, lipid oxidation and bacterial counts, while at the same time imparting a better appearance to the fresh lamb meat. In general, this effect was more pronounced at the higher level of TL (7.5%). High Pearson’s correlation coefficients were found between the sensory attributes, CIELAB coordinates and TBARS.     

Inhibition of aldehyde dehydrogenase enzyme by Durian (Durio zibethinus Murray) fruit extract

The scientific basis of the adverse, or at times lethal, effect of ingesting durian (Durio zibethinus Murray) while imbibing alcohol has not been established. Symptoms are reminiscent of the disulfiram–ethanol reaction (DER) arising from the inhibition of aldehyde dehydrogenase (ALDH). Cognizant of the inhibitory effect of sulphur compounds like disulfiram on ALDH and the rich sulphur content of durian, the influence of durian fruit extract on the ALDH-mediated oxidative metabolism of acetaldehyde was investigated. We report a dose-dependent inhibition of yeast ALDH (yALDH), at most 70% at 0.33 ppm (mg extract/l assay mix), by dichloromethane:pentane extracts. Sulphur-rich TLC fruit extract fractions that eluted farthest from the origin effected the greatest inhibitory action. yALDH assay using diethyl disulfide as internal standard further supports the role of durian’s sulfury constituents in the fruit’s ALDH-inhibiting property. Insight into the etiology of DER-like symptoms felt upon simultaneous durian and alcohol consumption is hereby presented.     

A laboratory assessment of the potential of different strains of the entomopathogenic fungi Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metschnikoff) to control Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) in stored cowpea

Twelve indigenous and exotic isolates of Beauveria bassiana and Metarhizium anisopliae were evaluated for their virulence and their ability to suppress populations of Callosobruchus maculatus in stored cowpea. LT₅₀ values ranged from 3.11 to 6.12 days following immersion in aqueous suspensions containing 1×10⁸ conidia ml⁻¹. Indigenous isolates that had been recovered from C. maculatus were more virulent in laboratory bioassays than exotic isolates from other insects. The two isolates with the shortest LT₅₀ values were compared in dose-response assays by immersion and by exposure to cowpea grains treated with dry conidia. In both assays B. bassiana 0362 was consistently more virulent than M. anisopliae 0351. By immersion, LC₅₀ values on day 6 post-treatment were 9.10×10⁴ and 7.10×10⁵ conidia ml⁻¹ for B. bassiana 0362 and M. anisopliae 0351, respectively. Exposure to treated grains gave LC₅₀ values of 1.15×10⁷ and 4.44×10⁷ conidia g⁻¹ grain for B. bassiana 0362 and M. anisopliae 0351, respectively. In 1 kg batches of cowpea stocked with 50 adult C. maculatus, B. bassiana 0362 at both 1×10⁷ and 1×10⁸ conidia g⁻¹ grain led to significant adult mortality and reduced F₁ emergence relative to untreated populations. At 1×10⁸ conidia g⁻¹ the effect of the fungus persisted into the F₁ generation. The net reproductive rates, R₀, measured 26 days after insects were released were 5.16 and 7.32 for the high and low doses compared to 9.52 for the untreated control.     

Insecticidal effect of Steinernema feltiae (Filipjev) (Nematoda: Steinernematidae) against Tribolium confusum du Val (Coleoptera: Tenebrionidae) and Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae) in stored wheat

The confused flour beetle, Tribolium confusum, and the Mediterranean flour moth, Ephestia kuehniella, are important pests of stored grain products. The insecticidal effect of three strains (UK 76 [=Nemasys], USA/SC, and Hawaii) of the entomopathogenic nematode Steinernema feltiae was determined in the laboratory, in wheat, against these pests. The nematodes were applied at three dose rates: 100, 300 and 900 nematodes/insect individual. The Hawaii strain was most virulent against T. confusum adults and larvae, with a significant dose effect in the case of the larvae. Larval mortality of this species reached 79% and 100% after 7 and 14 d of exposure to the nematodes, respectively, at the highest dose applied. On the other hand, adult mortality of T. confusum did not exceed 66%. In the case of E. kuehniella larvae, USA/SC performed best causing 52% and 69% mortality after 7 and 14 d exposure, respectively, at the highest dose tested. Since very few data are available on the effect of entomopathogenic nematodes against these pests, it is concluded that the Hawaii and USA/SC strains of S. feltiae should be further investigated as promising biological control agents for T. confusum and E. kuehniella.     

Characterisation of thermostable trypsin and determination of trypsin isozymes from intestine of Nile tilapia (Oreochromis niloticus L.)

Trypsin from intestinal extracts of Nile tilapia (Oreochromis niloticus L.) was characterised. Three-step purification – by ammonium sulphate precipitation, Sephadex G-100, and Q Sepharose – was applied to isolate trypsin, and resulted in 3.77% recovery with a 5.34-fold increase in specific activity. At least 6 isoforms of trypsin were found in different ages. Only one major trypsin isozyme was isolated with high purity, as assessed by SDS-PAGE and native-PAGE zymogram, appearing as a single band of approximately 22.39 kDa protein. The purified trypsin was stable, with activity over a wide pH range of 6.0–11.0 and an optimal temperature of approximately 55–60 °C. The relative activity of the purified enzyme was dramatically increased in the presence of commercially used detergents, alkylbenzene sulphonate or alcohol ethoxylate, at 1% (v/v). The observed Michaelis–Menten constant (K_m) and catalytic constant (K_cat) of the purified trypsin for BAPNA were 0.16 mM and 23.8 s⁻¹, respectively. The catalytic efficiency (K_cat/K_m) was 238 s⁻¹ mM⁻¹.     

Inhibitory effects of p-alkylbenzoic acids on the activity of polyphenol oxidase from potato (Solanum tuberosum)

In order to prevent potato from browning during preservation and processing, the effects of p-alkylbenzoic acids (p-propylbenzoic acid, p-butylbenzoic acid, p-pentylbenzoic acid, p-hexylbenzoic acid, p-heptylbenzoic acid, p-octylbenzoic acid) on the activity of potato polyphenol oxidase (PPO) have been studied. The PPO was extracted with PBS (pH 6.8), fractionated with ammonium sulphate, concentrated, and purified with Sephadex G-100 (1.8 × 100 cm) filtration chromatography. The active fractions were pooled and the PPO activity was determined to be 79.83 U/mg. The results showed that p-alkylbenzoic acids could strongly inhibit the activity of potato PPO. The values of IC₅₀ of these six inhibitors were determined to be 0.213, 0.180, 0.152, 0.106, 0.075 and 0.047 mM, respectively. Kinetic analyses showed that p-alkylbenzoic acids used in this experiment were reversible and noncompetitive inhibitors to the enzyme. The inhibitory effects were potentiated with increasing lengths of the hydrocarbon chains, indicating that the inhibitory efficiency on the enzyme was influenced by the steric effect of the substituted groups.     

Temperature-mediated functional response of Theocolax elegans (Hymenoptera: Pteromalidae) parasitizing Rhyzopertha dominica (Coleoptera: Bostrichidae) in stored wheat

The functional response of Theocolax elegans (Westwood) parasitizing the lesser grain borer, Rhyzopertha dominica (F.) was examined over a range of temperatures. A type II functional response equation was fitted to each temperature regime. The parasitization rate was highest at 30°C (20 hosts per day) and was lowest at 20°C (2 hosts per day). Handling time was inversely proportional to temperature, and ranged from 1.6 days at 20°C to 0.05 days at 30°C. Instantaneous search rate also changed with temperature. It was lowest at 20°C and highest at 30°C. A temperature-mediated functional response equation was fitted to the data, in which handling time was a quadratic function of temperature. The equation explained 74% of the variance in parasitization rate. Theocolax elegans has a narrower optimal temperature range than other parasitic stored-product Hymenoptera. Temperatures greater than 32.5°C caused high parasitoid mortality.     

Biology and management of Plodia interpunctella (Lepidoptera: Pyralidae) in stored products

Plodia interpunctella (Hübner), the Indian meal moth, is a world-wide insect pest of stored-products and processed food commodities. It can infest a variety of products and is perhaps the most economically important insect pest of processed food. In this review, we summarize the biology of P. interpunctella, discuss oviposition and development in relation to temperature, environment and food source, examine studies involving sampling and detection, describe various aspects of integrated control, summarize the current knowledge regarding management of P. interpunctella, and address potential areas for new research. The use of reduced-risk insecticides, non-chemical control, targeted pest management through spatial analysis and other means of identifying specific locations of infestations, and computer models that simulate population growth, are examples of some of those new areas of research.     

Augmented release of Teretrius nigrescens Lewis (Coleoptera: Histeridae) for the control of Prostephanus truncatus (Horn) (Coleoptera: Bostrichidae) in stored cassava chips

A trial was set up in northern Benin to evaluate the potential of Teretrius nigrescens to reduce the infestation and damage to cassava chips caused by storage insects. Cassava chips were stored for 5 months in mud silos and 50 adults of T. nigrescens were added when the stores were first filled. Stores where no predator was released were monitored as controls. The main storage insects observed were Prostephanus truncatus and Dinoderus spp. Initial chip weight varied between 102 and 246 g with no difference between treatments. Chip weight and number of holes on chips initially differed between treatments after 2 months of storage. After 3 months of storage, losses reached 40-50% without T. nigrescens and 30-40% when cassava chips were stored with T. nigrescens. A farmer can increase his profit by 1437 Fcfa/100 kg (1$=560 Fcfa, 1£=968 Fcfa; 1€=656 Fcfa, as on 2 December 2005) through the use of T. nigrescens because losses are reduced by 11%. Data analysis showed that there were significant differences (P<0.0001) between the two treatments for the number of holes, number of insects, weight of each chip as well as damage. There were twice as many P. truncatus and holes on chips in stores where T. nigrescens was not released. The addition of the predator to farmers’ stores is an economic option for controlling losses due to insects in cassava chips.