Control of Lipid Oxidation and Texture Changes in Frozen Sardine Mince by Combined Use of Milk Protein Concentrate and Caffeic Acid

The effectiveness of milk protein concentrate (MPC) and caffeic acid in controlling lipid oxidation and texture changes in 6-month frozen stored sardine mince was investigated using two different concentrations of caffeic acid (200 and 400 ppm) with and without MPC (4%). The extent of lipid oxidation was monitored by measuring thiobarbituric acid reactive substances (TBARS) while changes in the texture of mince due to frozen storage were assessed using Texture Analyzer (TA.XT Plus) and sensory analysis. Cooking loss upon frozen storage was also determined. The addition of MPC and/or caffeic acid significantly reduced lipid oxidation throughout the storage period. However, neither added benefit nor synergistic effect in retarding lipid oxidation was shown by caffeic acid. MPC not only retarded lipid oxidation but also moderated texture hardening, cooking loss, and moistness during frozen storage. No improvement by caffeic acid alone and no significant differences in sensory firmness and moistness between MPC-added mince with and without caffeic acid indicate that caffeic acid did not play any role in texture and moisture control. This study suggests that lipid oxidation and texture of frozen sardine mince can be better controlled by using MPC alone.     

A Study of Lipid Bioaccessibility in Canned Sardine (Sardina pilchardus) and Chub Mackerel (Scomber japonicus)

The lipid composition and bioaccessibility in canned sardine (Sardina pilchardus) and chub mackerel (Scomber japonicus) were studied. Triacylglycerols were absent from the bioaccessible fraction in both products and each FA had a different bioaccessibility level. While total fat bioaccessibility was in the 60–64% range, the bioaccessibility of ω6 PUFA and DHA was <55%. Bioaccessibility depended on canned fish product species: fat, EPA, DHA, and ω3 PUFA bioaccessibility levels seemed to be higher in canned sardine than in canned mackerel. Daily consumption of 278 g of canned mackerel or 47 g of canned sardine would ensure the recommended daily intake of EPA+ DHA.     

Effects of dietary levels of 20:5n-3 and 22:6n-3 on tissue lipid composition in juvenile Atlantic salmon, Salmo salar, with emphasis on brain and eye

Four dietary groups of juvenile Atlantic salmon, Salmo salar L., each with three replicates, were fed diets with increasing levels of docosahexaenoic acid (22:6n-3; DHA) and eicosapentaenoic acid (20:5n-3; EPA). Fatty acid composition of brain and eye was determined at the start and approximately every 3 weeks during the experimental period, and fatty acid composition of liver and fillet was determined in fish from the final sampling. Lipid class composition of brain and eye, and fatty acid composition of these lipid classes was determined at the end of the experiment. There was no effect of increasing dietary DHA content on fatty acid composition, lipid class composition or DHA levels in the lipid classes in the juvenile Atlantic salmon brain. The increasing dietary EPA content, however, was reflected in both the total fatty acid composition and in the EPA content in neutral lipids, phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol (PI). A minor effect of the increasing dietary DHA content was found in the lipid composition of the juvenile salmon eye. Both EPA and 18:2n-6 levels in eye, however, clearly reflected the increasing and decreasing, respectively, dietary levels of these two fatty acids. The dietary EPA levels also affected the EPA levels in neutral lipids, PC, PE, PI and PS (phosphatidylserine) in the juvenile salmon eye. The results demonstrate that these dietary levels of DHA had no effect on brain lipid composition and only a minor effect on eye lipid composition. Furthermore, the dietary EPA levels significantly affected the lipid composition of both brain and eye. The fillet fatty acid composition reflected the dietary fatty acid composition, except for the DHA/EPA ratio, which was reversed in fillet compared with that in the diets. The liver fatty acid composition was also affected by the increasing dietary EPA and DHA levels.     

Interactions between dietary docosahexaenoic acid and other long-chain polyunsaturated fatty acids on performance and fatty acid retention in post-smolt Atlantic salmon (Salmo salar)

A study with varying dietary inclusion levels (1, 5, 10, 15 and 20 g kg^?1) of docosahexaenoic acid (DHA; 22:6n-3) was conducted with post-smolt (111 ± 2.6 g; mean ± S.) Atlantic salmon (Salmo salar) over a 9-week period. In addition to the series of DHA inclusion levels, the study included further diets that had DHA at 10 g kg^?1 in combination with either eicosapentaenoic acid (EPA; 20:5n-3) or arachidonic acid (ARA; 20:4n-6), both also included at 10 g kg^?1. An additional treatment with both EPA and DHA included at 5 g kg^?1 (total of 10 g kg^?1 long-chain polyunsaturated fatty acids, LC-PUFA) was also included. After a 9-week feeding period, fish were weighed, and carcass, blood and tissue samples collected. A minor improvement in growth was seen with increasing inclusion of DHA. However, the addition of EPA further improved growth response while addition of ARA had no effect on growth. As with most lipid studies, the fatty acid composition of the whole body lipids generally reflected that of the diets. However, there were notable exceptions to this, and these implicate some interactions among the different LC-PUFA in terms of the fatty acid biochemistry in this species. At very low inclusion levels, DHA retention was substantially higher (~250 %) than that at all other inclusion levels (31–58 %). The inclusion of EPA in the diet also had a positive effect on the retention efficiency of DHA. However, EPA retention was highly variable and at low DHA inclusion levels there was a net loss of EPA as this fatty acid was most likely elongated to produce DHA, consistent with increased DHA retention with additional EPA in the diet. Retention of DPA (22:5n-3) was high at low levels of DHA, but diminished with increasing DHA inclusion, similar to that seen with DHA retention. The addition of EPA to the diet resulted in a substantial increase in the efficiency of DPA retention; the inclusion of ARA had the opposite effect. Retention of ARA was unaffected by DHA inclusion, but the addition of either EPA or ARA to the diet resulted in a substantial reduction in the efficiency of ARA retention. No effects of dietary treatment were noted on the retention of either linolenic (18:3n-3) or linoleic (18:2n-6) acids. When the total n-3 LC-PUFA content of the diet was the same but consisted of either DHA alone or as a combination of EPA plus DHA, the performance effects were similar.     

Oxidative stability of salmon and herring roe lipids and their dietary effect on plasma cholesterol levels of rats

ABSTRACT:   The oxidative stability of lipids from salmon roe and herring roe was compared with those of commercial fish oils originated from sardine and tuna. Both fish roe lipids contained high amounts of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Total EPA and DHA was more than 35% of total fatty acids in both roe lipids. On the basis of oxygen consumption, fish roe lipids showed the higher oxidative stability than both fish oils. This tendency in oxidative stabilities was also confirmed by the determination of propanal formation during oxidation. Analyses of lipid compositions suggested that the higher oxidative stabilities of fish roe lipids would be mainly due to the presence of phospholipids in them. Dietary effects of salmon and herring roe lipids were also determined. Little increase in total cholesterol level was observed in plasma lipids in rats fed salmon and herring roe lipids, although cholesterol content in fish roe lipids were 6.3% and 9.7% of total lipids for salmon roe and herring roe.     

Comparison of Proximate Composition and Fatty Acid Profile of On-Growing and Wild Mediterranean Horse Mackerel (Trachurus mediterraneus)

ABSTRACT

On-growing of horse mackerel is not known in the world. Recently, we have initiated on-growing of the Mediterranean horse mackerel in the Black Sea. Therefore, we aim to compare proximate composition and fatty acid profile of on-growing and wild horse mackerels to evaluate the effect on their nutritional value. Captured horse mackerels less than 13 cm were kept on-growing in sea cages and fed sea bass feed for a year in the southern Turkish Black Sea. Results showed seasonal variations in the proximate contents and fatty acid profile of both on-growing and wild fish groups (p < 0.05). Protein contents of the wild horse mackerel group were significantly higher than the on-growing mackerel group, while the opposite situation was observed for lipid contents (p < 0.05). Despite higher eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) levels (as fatty acid methyl esters %) of wild horse mackerel in comparison with on-growing group, much higher EPA + DHA contents were accounted for in on-growing fish in the edible portion resulting from higher lipid contents of these samples. The results suggest that lower amounts of fish meat, 50–90 g, from on-growing mackerel would cover the daily suggested value of EPA + DHA; this level is calculated as 51–150 g for wild fish meat.     

Optimizing the essential fatty acids, eicosapentaenoic and docosahexaenoic acid, in the diet of the prawn, Penaeus monodon

The dietary requirements of Penaeus monodon for eicosapentaenoic (20:5n‐3; EPA) and docosahexaenoic (22:6n‐3; DHA) acids were examined. These requirements were examined when dietary levels of linoleic (18:2n‐6; LOA) and linolenic acids (18:3n‐3; LNA) were also provided at previously established optimal levels of 14 and 21% respectively of the total lipid fatty acids. A 5 × 5 factorial design was used with incremental amounts (0, 4, 8, 12 and 16% of total fatty acids) of EPA and/or DHA. An additional diet containing cod‐liver oil was provided as a reference diet. The total lipid content of all of the 25 treatments and reference diets was maintained at the same level of 75 g kg^?1. Growth of prawns fed with the reference diet after 50 days was 244 ± 21%. The greatest response to singular additions of EPA or DHA was with a 12% inclusion of either fatty acid, resulting in 287 ± 21 and 293 ± 18% weight gain, respectively. Growth was generally better when combinations of EPA and DHA were used, the optimal combination being EPA 4% and DHA 4%, resulting in 335 ± 25% weight gain. Addition of high levels of either of the highly unsaturated fatty acids (HUFA) in the diet had a negative effect on growth. Digestibilities of the total neutral lipid and specific fatty acids were examined during the growth trials. The digestibility of total neutral lipid was usually higher when either or both HUFA were present, however there were few significant differences between treatments that contained either or both HUFA. Following the growth trials, digestive glands (DG) of prawns fed with the various diets were analysed to determine the total lipid content and fatty acid composition. Total lipid in the digestive gland increased with the inclusion of DHA, but was not significantly affected by the addition of EPA. The fatty acid composition of the digestive gland lipid generally reflected that of the diet. However, the maximum retention of EPA (11.1% of total DG fatty acids) and DHA (10.7% of total DG fatty acids), was not directly proportional to the amount of either fatty acid present in the diet. These results demonstrate that both EPA and DHA have considerable growth promoting capacity. This growth promoting capacity is enhanced when an optimal balance of both fatty acids are incorporated into the diet.     

An Invasive Shrimp Species,Machrobrachium nipponense,in Anzali Wetland Demonstrated a Potential Source for Commercial Fishing

ABSTRACT

The proximate content, fatty acids composition, and nutritional quality index (NQI) of Macrobrachium nipponense at three habitats in the Anzali wetland in Iran were investigated as a potential source for human consumption. The highest amounts of protein, lipid, ash, and energy contents in muscle of M. nipponense were showed in autumn (non-reproductive season) (p < 0.05). The main monounsaturated fatty acids (MUFA) were oleic acid (C18:1 n9 C, C18:1 ω9 T) and palmitoleic acid (C16:1). Moreover, the main polyunsaturated fatty acids (PUFA) were docosahexaenoic acid (DHA, C22:6 n3), eicosapentaenoic acid (EPA, C20:5n3), arachidonic acid (ARA, C20:4 n6), linoleic acid (LA, C18:2 ω6), and α-linolenic acid (ALA, C18:3 n3). The predominant individual saturated fatty acid (SFA) was palmitic acid (0.07–13.4%), while oleic acid (14.7–26.3%), EPA (3.5–12.7%) and linoleic acid (0.04–14.9%) represented the most abundant individual MUFA and PUFA in M. nipponense. The highest mean value of EPA+DHA (14.0), n3/n6 (1.02), ΣMUFA/ΣSFA (1.05), ΣPUFA/ΣSFA (1.04), and EPA/DHA (3.8) ratios in M. nipponense was in autumn. The range of atherogenicity index (AI) and thrombogenicity index (TI) was much lower, from 0.42 to 0.6 and from 0.33 to 0.57, respectively, in terms of season. The results obtained in the present study show that M. nipponense is an excellent nutritional food source in the Anzali wetland.     

Influence of egg lipids and fatty acids on egg viability, and their utilization during embryonic development of spotted wolf-fish, Anarhichas minor Olafsen

The changes in egg lipids and fatty acid compositions that occur during embryonic development of spotted wolf‐fish, Anarhichas minor, were examined by monitoring individual egg batches from the time of spawning (egg stripping) until hatching. The lipids, present as 3.7±0.1% of the wet mass of the freshly stripped eggs, contained high percentages of monoenes (monounsaturated fatty acids (MUFAs), ca. 33%) and polyenes (ca. 43%) and approximately 20% saturated fatty acids (SFAs). The fatty acid profiles were dominated by a small number of fatty acids. The major SFA was 16:0 (ca. 14%), the dominant MUFA was 18:1 n‐9 (ca. 21%), and among the polyenes, the n‐3 highly unsaturated fatty acids (HUFAs) 22:6 n‐3 docosahexaenoic acid (DHA) and 20:5 n‐3 eicosapentaenoic acid (EPA) were present in the highest concentrations (EPA, ca. 16%; DHA, ca. 19%). The n‐6 HUFA 20:4 n‐6 arachidonic acid (AA) was present as ca. 1% of the total fatty acids in the freshly stripped eggs. This resulted in an AA:EPA of ca. 0.07, which is lower than reported for eggs of many other fish species. As embryonic development progressed, the percentage contribution of AA to the total fatty acids almost doubled. There were also increases in the relative proportions of SFAs (due mainly to an increase in the percentage of 16:0 to ca. 16% at hatch) and DHA (to ca. 23%), and there was a corresponding decrease in the percentage of MUFAs (mostly brought about by a decrease in the percentage of 18:1 n‐9 to ca. 18% at hatch). The most marked changes occurred towards the end of incubation. The percentage of EPA changed little during incubation. This implies that there was selective retention of DHA, 16:0 and AA, and these fatty acids were probably incorporated into cell membranes. MUFAs, particularly 18:1 n‐9, seem to have been catabolized to provide energy for the developing embryo, and some EPA also seems to have been utilized as an energy source. Survival of eggs to the eyed stage (range ca. 10–80%) and to hatch (ca. 5–75%) was negatively correlated with the %AA, %EPA and AA:DHA of the freshly stripped eggs. There was also a negative correlation between AA:EPA and egg survival, which implies that there is not a universal requirement for a high AA:EPA to ensure high rates of survival of fish eggs.     

Chemical Composition of Fresh and Salted Lumpfish (Cyclopterus lumpus) Roe

The original "Eskimo" cardiovascular health benefit of omega-3 fatty acids was ascribed to EPA (eicosapentaenoic acid). The two readily available fish body oils (from U.S. menhaden and Japanese sardines) have approximately 180 mg of EPA/gram, accompanied by much less (120 mg) DHA (docosahexaenoic acid). The EPA was recognized as the source of a prostaglandin (PGI₃) acting as an agent to keep the vascular walls soft and flexible, thus hindering atherosclerosis and vascular constriction, but DHA presented a problem as no such role for it was known. Australian researchers identified DHA as preventing cardiac arrhythmia because they used tuna body oil (DHA ~5 x EPA) in their research. Generally our analyses of salmon muscle triglycerides seem to show DHA > EPA. Total salmon muscle lipids in fillets obtain extra DHA from phospholipids. In the United Kingdom an official medical body now recommends eating fish twice a week and "oily" fish once a week. Examination of DHA for a legitimate health claim in the cardiovascular system, and in infant neural developments and nutrition, and maternal nutrition, offers an alternative market exploiting the most recent developments. These matters should be investigated by the salmon aquaculture industry.     

Effects of dietary <Emphasis Type="Italic">n</Emphasis>-3 fatty acids on Toll-like receptor activation in primary leucocytes from Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>)

The shortage of the n-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on the international markets has led to increasing substitution of fish oil by plant oils in Atlantic salmon (Salmo salar) feed and thereby reducing the EPA and DHA content in salmon. However, the minimum required levels of these fatty acids in fish diets for securing fish health are unknown. Fish were fed with 0, 1 or 2% EPA or DHA alone or in combination of both over a period, growing from 50 to 400 g. Primary head kidney leucocytes were isolated and stimulated with Toll-like receptor (TLR) ligands to determine if EPA and DHA deficiency can affect expression of important immune genes and eicosanoid production. Several genes related to viral immune response did not vary between groups. However, there was a tendency that the high-level EPA and DHA groups expressed lower levels of IL-1β in non-stimulated leucocytes. These leucocytes were also more responsive to the TLR ligands, inducing higher expression levels of IL-1β and Mx1 after stimulation. The levels of prostaglandin E2 and leukotriene B4 in serum and media from stimulated leucocytes were lower in both low and high EPA and DHA groups. In conclusion, leucocytes from low EPA and DHA groups seemed to be less responsive towards immunostimulants, like TLR ligands, indicating that low levels or absence of dietary EPA and DHA may have immunosuppressive effects.     

Antioxidant Properties of Chitosan Coatings on Frozen Atlantic Salmon Fillet Portions

Atlantic salmon contain omega-3 fatty acids, which play important roles in promoting human health but are highly susceptible to oxidation. Chitosan has been shown to have antioxidant properties which could be beneficial in extending the shelf life of Atlantic salmon; however, the effects of chitosan molecular size on oxidation of salmon fillets have not been reported. The primary objective of this study was to evaluate the effects of chitosan coatings on lipid oxidation of Atlantic salmon fillet portions during 5 months frozen storage. The effects of chitosan molecular weight (high molecular weight, low molecular weight, and enzymatically degraded chitosan), concentration (0.5 and 1.0%), and the addition of 1% ascorbic acid to the chitosan coating were evaluated. Chitosan molecular weight significantly affected oxidation as evaluated by propanal levels and changes in L* values of the fillet portions. The chitosan treatment with added ascorbic acid resulted in the lowest propanal levels during 5 months of frozen storage. Manipulation of chitosan molecular weight and incorporation of natural antioxidants into chitosan coatings can provide an effective method of reducing lipid oxidation during extended frozen storage of seafood products.     

Effect of a Supplemented Diet with Canned Sardine on the Lipid Fraction of Human Plasma and Erythrocytes

Abstract

Over the past 20 years many studies and clinical investigations have addressed the beneficial effects of polyunsaturated fatty acids in general and ω3 fatty acids in particular. It is currently known that ω3 fatty acids play important role in the prevention of several pathologies like the coronary heart disease, hypertension, diabetes and other. The protection played by ω3 fatty acids against coronary heart disease may be related to antiatherogenic effects and/or to the modification of risk factors through mechanisms related to lipid metabolism. The objective of this work was to evaluate, in healthy volunteers, the effect of a diet supplementation with canned sardine, which is rich in co3 fatty acids on the profile of plasma and erythrocytes, lipids, phospholipids and fatty acids.

The serum levels of total cholesterol and high density lipoproteins (HDL) did not change during the period of the supplementation, tri-acylglycerol levels showed a trend for a decrease and low density lipoproteins (LDL)-derived thiobarbituric acid-reactive substances (LDL TBARS) levels showed a decrease. Incorporation of eicosapentaenoic and docosahexaenoic fatty acids (EPA and DHA, respectively) into plasma and erythrocyte phospholipids was observed. The results suggest that the undergone dietary supplementation may have allowed the incorporation of co3 polyunsaturated fatty acids into cell membranes and has increased the resistance of LDL to oxidative stress. This may represent protection factors against atherosclerosis and cardiovascular disease.     

Effect of Liquid Smoke Dipping and Packaging Method on the Keeping Quality of Raw and Cooked Chub Mackerel (Scomber japonicus) Fillets

Changes in biochemical and sensory attributes of chub mackerel (Scomber japonicus) fillets during storage treated by immersion in a 12% NaCl brine with or without 0.2% liquid smoke were investigated. Brine-treated chub mackerel fillets were packaged in modified atmosphere or under vacuum and were stored at 4°C. The brined fillets were stored and evaluated raw and after heat treatment as cooked samples. Fish fillets were assessed organoleptically to determine acceptability and by measuring the pH, oxidation levels (TBARS test), and total volatile basic nitrogen (TVBN). Fillets treated in brine with liquid smoke exhibited similar pH and significantly lower TBARS and TVBN values (p < 0.05). The combination of the use of brines containing liquid smoke and subsequent vacuum packaging produced the lowest TBARS and TVBN values and resulted in extended acceptability.     

High dietary 18:2n‐6/18:3n‐3 ratio does not inhibit elongation and desaturation of 18:3n‐3 to EPA and DHA in Atlantic salmon (Salmo salar L.)

This study investigated whether retention of n‐3 fatty acids (FAs) is influenced by the levels of dietary monounsaturated (MUFA)‐, saturated (SFA)‐ and n‐6 FAs. The feeding trial used a mixture design, with rapeseed‐, palm (PO)‐ and soybean oil (SO), providing high levels of MUFA, SFA and n‐6 FAs, respectively, while 18:3n‐3, eicosapentaenoic acid (20:5n‐3) and docosahexaenoic acid (DHA, 22:6n‐3) were kept constant. Furthermore, a diet group with high SO and reduced 18:3n‐3 was included in addition to the mixture design. There were no differences in growth or proximate composition, but the PO diet gave reduced FA digestibility and increased feed conversion ratio. All diet groups had a net production of DHA, shown by retention values >100% (133%–177%). High dietary n‐6 FAs of up to 43% of FAs (18:2n‐6/18:3n‐3 ratio 8.6) did not negatively affect DHA retention, but rather had a small positive impact. High levels of substrate (dietary 18:3n‐3 at ~10% of FAs) did not enhance DHA retention compared to 5% 18:3n‐3. This is a highly significant finding for the aquaculture industry, allowing for greater flexibility in the choice of lipid sources to replace fish oil in salmon feeds.     

Effects of dietary vegetable oils and varying dietary EPA and DHA levels on intestinal lipid accumulations in Atlantic salmon

High dietary content of vegetable oil (VO) has been associated with increased intestinal lipid accumulations in fish. The extent of this in aquacultured Atlantic salmon (Salmo salar L.) and its health effects are not certain. Samples were therefore collected from two separate feeding trials to investigate the effect of high dietary VO on intestinal lipid accumulations in Atlantic salmon. In the first trial, the fish were fed diets high in plant protein and with fish oil or ~80% of the fish oil replaced with either olive oil, rapeseed oil or soybean oil in a land‐based experimental set‐up. The second trial was performed in sea cages under commercial production conditions, and the fish were fed two dietary concentrations of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (9.7% or 5.5% EPA + DHA of total fatty acids). Neither dietary VO nor variations in EPA and DHA led to any significant effects on intestinal health or lipid accumulations. There were, however, indications of a delayed lipid transport in the rapeseed oil‐fed fish of the first trial, possibly caused by high dietary ≥18‐carbon fatty acids and low dietary 16:0 fatty acid and cholesterol.     

Effect of dietary ω3 and ω6 fatty acids on growth and feed conversion efficiency of coho salmon (Oncorhynchus kisutch)

Sixteen isocaloric diets varying only in the levels of ω3 and ω6 fatty acids were prepared. Each diet was fed to duplicate groups of coho salmon for 14 weeks. At termination, the final weight of each diet group fish was determined. The optimum level of dietary ω3 fatty acid ranged from 1% to 2.5%. Dietary ω6 fatty acid higher than 1% depressed fish growth. Fish growth, feed conversion efficiency, fish mortality and fatty acid composition of fish phospholipids were determined.     

Fatty Acid Composition of U.S. West Coast Albacore Tuna (Thunnus alalunga) and the Effects of Canning and Short-Term Storage

ABSTRACT

Changes in the fatty acid composition of 13 bled and blast-frozen U.S. West Coast albacore tuna (Thunnus alalunga) were determined following custom-canning and 5 weeks of canned storage. In accordance with typical custom-canning procedures, no additional packing materials were used, and the product did not undergo a precooking step. Raw albacore tuna was found to be rich in n-3 polyunsaturated fatty acids (PUFAs), with an average of 9.1% (% wt of total fatty acids) EPA and 33.8% DHA. Canning and short-term storage did not cause major fluctuations in the fatty acid profiles of the albacore tuna, with no significant changes in the total fractions of saturated fatty acids (30–31%), monounsaturated fatty acids (19%), PUFAs (50%), or n-3 PUFAs (46–47%). On a tissue weight basis, U.S. West Coast albacore tuna was found to contain high levels of total n-3 PUFAs, with 3.3 g/100 g tissue in raw tuna and 4.0–4.1 g/100 g tissue in products that were canned and stored for 5 weeks.