Effect of bleeding on the quality of amberjack Seriola dumerili and red sea bream Pagrus major muscle tissues during iced storage

The effect of bleeding on white muscle quality in amberjack and red sea bream was evaluated by measuring ATP-related compounds, volatile basic nitrogen (VBN), and the trimethylamine (TMA) content. The freshness was evaluated by the K value, and the degree of spoilage was elucidated by VBN and TMA. ATP was rapidly converted to ADP and AMP in the muscle and IMP was the main product of ATP degradation during iced storage. For both species, the IMP content was higher in the muscles of fish that were bled than in those unbled during iced storage. Conversely, the K value and the levels of hypoxanthine (Hx) and VBN were higher in the muscle tissue of unbled fish than in the bled tissue of both species. Similarly, the TMA content was higher in the unbled muscle tissues of both species after a week of storage.     

Texture and Quality Assessment of Ready-to-eat Farmed Obscure Puffer Fish (Takifugu obscurus) Fillet by Evaluating Bacterial and Myofibrillar Degradation and Biochemical Changes during Refrigerated Storage

ABSTRACT

Obscure puffer fish (Takifugu obscurus) flesh, often eaten as raw fish fillet, is susceptible to softening. To investigate the mechanisms for the softening of flesh and changes in firmness, the proliferation of total microbial flora, Enterobacteriaceae, Pseudomonas spp., and lactic acid bacteria (LAB), total volatile basic nitrogen (TVBN), trimethylamine (TMA), and pH were periodically assayed during storage at 0, 4, and 8ºC. Myofibril extraction and separation of puffer fish flesh was carried out and the length of the separated myofibrils was measured periodically using an optical microscope during the entire storage time. Transmission electron microscopy was also used to inspect the changes in myofibrillar characteristics of the fish flesh at the beginning and end of the storage period. The results indicated that lower chilled temperature can contribute to reduced proliferation of microbial flora and less variation in TVBN, TMA, and pH and further maintain the integrity of myofibrils and retain firmness of puffer fish flesh during storage.     

Effects of Bicarbonate,Xanthan Gum,and Preparation Methods on Biochemical,Physicochemical, and Gel Properties of Nile Tilapia (Oreochomis niloticus Linn) Mince

This study was aimed to determine the effects of phosphate compound substitutions (sodium bicarbonate and xanthan gum) and preparation methods—headed, gutted whole fish, and mince; fresh and after frozen storage (?20°C for 3 months)—on Nile tilapia mince qualities. Results showed that bicarbonate (0.3% with 8% sucrose/sorbitol) is an efficient phosphate compound replacement as evidenced by the comparative values of salt extractable protein, Ca²⁺-ATPase activity, total sulfhydryl content, and textural properties to those of the phosphate-added—0.3% sodium tripolyphosphate (STPP) with 8% sucrose-sorbitol—sample after 3-month frozen storage (p > 0.05). Both cryoprotected samples containing STPP or bicarbonate exhibited higher denaturation temperatures of myosin than others. Xanthan gum (0.5%) could neither stabilize the biochemical and physicochemical properties of mince during 3-month frozen storage nor improve textural properties of gel from frozen whole fish.     

Changes in Biochemical Compositions and Quality of White Shrimp (Litopenaeus vannamei) During Storage

ABSTRACT

To understand biochemical characteristics, storage stability, and freshness indicators of white shrimp (Litopenaeus vannamei), changes in extractable nitrogenous compounds, microbial count, and sensory rating of white shrimp during storage at 25 and 4°C were investigated. Free amino acids showed a slow increase during storage at 25°C, but no obvious change was found at 4°C. Adenosine triphosphate (ATP) and adenosine diphosphate (ADP) were found at initial stage and decreased rapidly after storage. Both inosine 5′-monophosphate (IMP) and adenosine monophosphate (AMP) increased and then decreased during storage. Inosine, hypoxanthine, and the K-value gradually increased with time. The levels of total volatile basic nitrogen (TVB-N), NH₃, and trimethylamine (TMA) of white shrimp increased with storage time at 25 and 4°C. The TVB-N, NH₃, TMA, inosine, hypoxanthine, and K-value could be considered as freshness indicators of white shrimp during storage. However, the total plate count did not corroborate the acceptability recommended limits for white shrimp during storage. The sensory evaluation, associated with TVB-N, TMA, and K-value, showed the quality was unacceptable after 6 h storage at 25°C and 7 days at 4°C.     

北极虾在不同冷藏条件下的质量变化与质量指标

采用感官评估、物理、化学、微生物分析方法,对贮藏在不同冷藏条件下的北极虾的质量变化与质量指标进行了研究。保藏方法分别为传统冰块冷却法、液态冰冷却法、盐水-冰混合冷却剂法,贮藏环境温度为-1．5℃或1．5℃。研究结果表明,液态冰冷藏法对延缓虾腐败变质的效果比传统冰块冷藏法、盐水-冰混合冷却剂冷藏法好。经过1d的贮藏后发现,无论在-1．5℃还是1．5℃的环境温度下,保藏在液态冰中虾的挥发性总氮(TVB-N)都有所下降,并且在-1．5℃的环境温度下,其TVB-N值在3d后才回升到刚开始贮藏时的水平,而在其它保藏方法中,虾的TVB-N值始终随贮藏时间的延长而增加;除保藏在液态冰-1．5℃环境中虾的三甲胺(TMA)在贮藏的第1天有所降低以外,其它各试验组虾的TMA值均随贮藏时间的延长而增加。检测结果还表明,保藏在传统冰块及盐水-冰混合冷却剂中虾的细菌总数(TVC)增加最快,其次是保藏在液态冰中1．5℃的环境下。而保藏在液态冰中-1．5℃的环境下,虾的细菌繁殖最慢,并且在贮藏初期,产生了明显的细菌总数降低及细菌繁殖被抑制的现象。采用主要因子分析(PCA)和方差分析(ANOVA)发现,质量指标如TVB-N、TMA、TVC、pH、NH3(电子鼻测量值)和感官评估结果等相互之间存在着良好的相关性。     

Effect of Thermal Microbial Inactivation and Washing on Quality Properties of Fish Sausage During Cold Storage (4°C)

The effect of microbial inactivation and washing process on physicochemical and sensory quality of pasteurized fish sausage during cold storage (4°C) was evaluated. Mince was obtained after the grinding; washed mince (WM) was obtained by single washing of mince; and surimi was obtained after three washing and dewatering cycles. Texture profile analysis results demonstrated that textural characteristics of pasteurized fish sausages remain stable during storage period (p > 0.05). Color analysis showed that Hunter L* had an increasing trend, and a* and b* had a decreasing trend over time (p < 0.05). Thus, it is possible to inactivate all microorganisms in pasteurized fish sausage without any undesirable effect on physicochemical and sensory quality.     

Using Antioxidants for Extending the Shelf Life of Frozen Nile Karmout (Claries lazera) Fish Mince

Abstract

Nile karmout (Claries lazera)fish mince was treated with 0.5% sodium tripolyphosphate (STPP), 0.5% ascorbic acid, 0.5% citric acid, and 0.1% Na EDTA, then stored at -18°C for 6 months. Yield, proximate composition, thiobarbitunc acid value (TBA), free fatty acid (FFA), salt soluble protein (SSP), non-protein nitrogen (NPN), water holding capacity (WHC), and cooking loss were determined at 0, 2, 4, and 6 months of storage. The yield of skinned fillet from whole fish was 47.0%. Moisture, protein, fat and ash offish mince were 72.9%, 17.6%, 8.5% and 1.0%, respectively. TBA values and FFA levels indicated that ascorbic acid and Na₂EDTA were the most effective antioxidants. Samples treated with STPP had the highest salt soluble protein. STPP was effective in maintaining the WHC of treated sample. Samples treated with citric acid had the highest cooking loss. Results of the study suggested that karmout is an excellent source for producing fish mince for further processing. Samples treated with antioxidants retarded the rancidity during frozen storage.     

Autolysis and Characterization of Sarcoplasmic and Myofibril Associated Proteinases of Oxeye Scad (Selar boops) Muscle

ABSTRACT

The autolytic profile of oxeye scad mince was characterized. Mince showed higher proteolytic activity than washed mince. The highest autolysis was observed at 65 and 60°C for mince and washed mince, respectively. Both mince and washed mince showed the optimum pH for autolysis at pH 9.0, and their activities decreased with increasing NaCl concentration (0–3.5%). Autolysis of washed mince was markedly inhibited by soybean trypsin inhibitor (SBTI), suggesting that myofibril-associated proteinase was serine proteinase. Sarcoplasmic proteinase was characterized to be heat-activated alkaline proteinase having the optimal pH and temperature of 9.0 and 60°C, respectively. The activities were stable at pH range of 8.0–11.0 at 20–40°C. The crude proteinase was inhibited by N-p-tosyl-L-lysine chloromethyl ketone, SBTI, and phenylmethanesulfonyl fluoride, suggesting the predominance of serine proteinases, especially trypsin. NaCl suppressed the activity while β-mercaptoethanol, dithiothreitol, and CaCl₂ activated the activity. Therefore, trypsin-like proteinase is a major endogenous proteinase responsible for autolysis in oxeye scad muscle. The present results can be used as scientific guidelines to predict the gel strength of surimi made from oxeye scad muscle.     

Gelation Characteristics of Mince and Washed Mince From Small-Scale Mud Carp and Common Carp

Small-scale mud carp (SC, Cirrhina microlepis) and common carp (CC, Cyprinus carpio) are underutilized freshwater fish. The objective of this study was to elucidate the gel-forming ability of mince and washed mince from these species in relation to washing cycles and CaCl₂ addition. Protein loss of up to 67% was observed after three-washing cycles of both species, rendering relatively low yield. About 85–90% of total fat was removed from SC flesh after three-washing cycles, but only two-washing cycles were sufficient for fat removal in CC mince. Mince and washed mince of both species did not exhibit severe proteolysis with low autolytic activity at 65°C. SC exhibited superior gel-forming ability to CC. Two- and three-washing cycles in conjunction with a 40°C-preincubation resulted in the highest textural properties of SC and CC, respectively. CaCl₂ increased the breaking force of CC and SC mince gels up to 65 and 95% at 0.3 and 0.5% CaCl₂, respectively, as compared to without CaCl₂. However, it showed negative effects on surimi gels of both species. Both SC and CC are potential freshwater fish resources for mince and washed mince production with reasonable good gel-forming ability.     

Effects of Modified Atmospheric Packaging Conditions on the Quality Changes of Pasteurized Oyster (Crassostrea belcheri) Meat during Chilled Storage

ABSTRACT

The effects of different atmospheric conditions on the quality and the shelf life of pasteurized oyster meat in chilled storage were investigated. The alternatives tested for pasteurized oyster meat were normal air (Control), 75% CO₂/25% N₂ (MAP-1), 75% CO₂/25% O₂ (MAP-2), 75% CO₂/5% O₂/20% N₂ (MAP-3), and vacuum packing (VP). The various observations of microbiological (total viable count (TVC), Enterobacteriaceae, Pseudomonas spp., lactic acid bacteria (LAB), and hydrogen sulfide–producing bacteria), physical (color coordinates L*, a*, and b*, and cutting strength), chemical (moisture, pH, total volatile basic nitrogen (TVB-N), peroxide value (PV), and thiobarbituric acid reactive substances (TBARS)) and sensory properties were performed every 3 days over a total of 27 days. During storage across all the treatments, L*, a*, cutting strength, pH, moisture content, and sensory scores decreased; whereas, b*, TVB-N, PV, TBARS, and microorganisms increased. However, among the various alternatives, MAP-1 had the best retention of oyster quality during storage. Based on sensory and microbiological qualities, the shelf lives in chilled storage were 9 days for control, 15 days for VP, 21 days for MAP-2 and MAP-3, and 24 days for MAP-1-treated cases.     

Effects of High Pressure Treatment on Physicochemical Quality of Pre- and Post-Rigor Palm Ruff (Seriolella Violacea) Fillets

Fish and fish products are characterized for having a short shelf life. Nonthermal processing techniques such as high hydrostatic pressure (HHP) have increasingly been employed to extend shelf life of food products. The aim of this study was to evaluate changes on flesh physicochemical spoilage parameters (pH, total volatile bases (TVB-N), trimethylamine (TMA), thiobarbituric acid (TBA), and color) of palm ruff (Seriolella violacea) fillets in pre- and post-rigor conditions, subjected to two different HHP conditions: 450 MPa and 550 MPa, for 3 and 4 min each. Unpressurized and pressurized fillets were kept in chilled storage (4 ± 1°C) for 26 days to assess the effect of HHP on shelf life. pH and TBA values increased after HHP treatment and with storage time for both unpressurized and pressurized samples. This is attributable to pressure-induced lipid oxidation. Lightness (L*) values increased with pressure, where fish fillets had a cooked appearance. TMA and TVB-N values decreased after HHP treatment compared to the unpressurized samples, showing that HHP treatment is an efficient method to maintain the quality of palm ruff fillets. There was no clear difference between pre- and post-rigor in the parameters evaluated.     

Quality Changes and Browning Developments During Storage of Dried-Seasoned Squid (Dosidicus gigas and Ommastrephes bartrami)

ABSTRACT

The physicochemical changes—moisture, water activity, crude protein, total volatile basic nitrogen (TVB-N), trimethylamine oxide (TMAO), dimethylamine (DMA) and formaldehyde, reducing sugar and amino acid; microbiological changes—aerobic plate count (APC) and coliforms; and browning development (browning intensity and color) in dried-seasoned squids (Dosidicus gigas and Ommastrephes bartrami) during storage at 25°C were investigated. After the storage, the moisture and water activity decreased significantly. The content of crude protein decreased, accompanied by progressive accumulation of TVB-N and slow increase of APC. The high level of TMAO in dried-seasoned squid gradually decreased, whereas DMA and formaldehyde increased significantly during the whole period of storage. The browning intensity and b* value significantly increased, which revealed the occurrence of browning in two squid products. The dominant reducing sugar, lactose, and glucose, and the total amino acid including lysine and proline, decreased remarkably, which were well-correlated with the browning development. The quality deterioration in jumbo squid was more rapid than in neon flying squid partially due to the high level of reducing sugar in the former product. To improve quality and resist browning reaction of dried squid product during storage, the addition of extrinsic reducing sugar became a control factor.     

Combined Effect of Ozonized Water Pretreatment and Ozonized Flake Ice on Maintaining Quality of Japanese Sea Bass (Lateolabrax japonicus)

The effect of the combined application of ozonized water pretreatment and ozonized flake ice (O+OIce) as compared with traditional flake ice (CK), ozonized flake ice (OIce), and ozonized water pretreatment and traditional flake ice (O) was investigated for the storage of Japanese sea bass. Microbiological, chemical, and sensory analysis and moisture content were carried out throughout a storage period of 18 days. A slower increase of total viable mesophilic count (TVC), total volatile base nitrogen (TVB-N), and thiobarbituric acid reactive substances (TBARS) was observed for O+OIce as compared with OIce, O, and CK. Treatment of O+OIce also allowed a better control of pH as compared with OIce, O, and CK. None of the treatments resulted in a significant difference in myofibrillar protein extractability and moisture content. The shelf life of Japanese sea bass treated with CK, OIce, O, and O+OIce, determined by sensory evaluation, was 9, 15, 12, and more than 18 days, respectively. Principal component analysis results showed that the treatments significantly affected the fish quality during storage, and the sensory attributes had a significant correlation with TVB-N, TVC, and TBARS. In conclusion, the combined application of ozonized water and ozonized flake ice had the best effect on fish quality maintenance.     

Biochemical,Physical, Chemical,and Microbiological Assessment of Blue Shrimp (Litopenaeus stylirostris) Stored in Ice

Postmortem changes in blue shrimp (Litopenaeus stylirostris) muscle were studied on the basis of biochemical, chemical, physical, and microbiological changes during an 18 day storage period at 0°C. Adenosine 5′-triphosphate (ATP) content, breakdown products, K-value, pH, trimethylamine nitrogen (TMA-N), total volatile basic nitrogen (TVB-N), water holding capacity (WHC), color, and texture (shear force) changes were examined. Also, total mesophilic and psychrophilic bacterial counts were measured. K-value increased linearly (r² = 0.98) from an initial value of 1.37 ± 0.59 to 59.42 ± 6.05% at Day 18. Spoilage indicators TVB-N and TMA-N increased from 29.56 ± 1.33 and 0.69 ± 0.25 to 39.04 and 2.04 ± 0.59 mg of N/100 g at Day 18, respectively; meanwhile, the total viable counts of mesophilic and psychrophilic bacteria increased from 3.48 ± 0.44 and 2.61 ± 0.29 log CFU/g to 6.27 ± 0.21 and 7.14 ± 0.39 log CFU/g, respectively, which indicated spoilage at the end of the storage period. The pH, texture, WHC, and color were affected (p < 0.05) during the storage period. Overall, results indicate that blue shrimp muscle quality was maintained for 12 days of storage in ice.     

Effect of Delayed Icing on the Microbiological,Chemical, and Sensory Properties of Caspian Sea Golden Grey Mullet (Liza aurata)

Research has shown that fresh fish is an extremely perishable food as compared to other food commodities. In this study, quality deterioration during storage of golden grey mullet (GGM) iced for 0, 6, and 12 h after landing was monitored. Microbiological indices (total viable count, TVC; psychrophilic counts, PTC; lactic acid bacteria, LAB; and Enterobacteriaceae counts, EBC), chemical parameters (pH; total volatile basic nitrogen, TVB-N; peroxide value, PV; thiobarbituric acid, TBA; and free fatty acids, FFA) values increased over the preicing duration. Delayed icing did not lead to significant increases (p > 0.05) in TVC, PTC, LAB, and EBC throughout the period of storage. All chemical indices increased during storage (p < 0.05). This study showed that sensorial analysis of GGM correlated well with microbiological analysis. Results of this study based on the microbiological and raw fish sensorial data indicated that the shelf life of GGM stored in ice immediately after catch was approximately 10–11 days, while the 6- and 12-h delays in icing shortened the shelf life to approximately 6–7 and 2–3 days, respectively.     

Quality attributes,polycyclic aromatic hydrocarbon,and heavy metal profile of traditional drum-smoked Guinean barracuda fish from Lagos State,Nigeria

This study was carried out to assess the quality attributes, polycyclic aromatic hydrocarbon (PAH), and heavy metal profile of traditional drum-smoked Guinean barracuda fish from Lagos State, Nigeria. Fresh Guinean barracuda fish (60) samples were obtained from 20 different fish-processing centers. The fresh fish samples were smoked with traditional drum kiln. The fresh and smoked fish samples were analyzed for proximate composition, quality indices (peroxide value (PV), thiobarbituric acid (TBA), total nitrogen base-nitrogen (TVB-N), trimethylamine acid (TMA), and free fatty acid (FFA)), PAH, heavy metals, and microbiological (coliform, fungi, Listeria monocytogenes (LM), Salmonella paratyphi (SP), and Staphylococcus aureus (SA)) analyses. The results revealed that the mean moisture, protein, fat, crude fiber, ash, and carbohydrate contents of traditional drum-smoked fish samples were in the range 10.93–14.15%, 51.97–60.96%, 11.34–15.99%, 1.74–2.99%, 1.12–1.56%, and 7.70–17.99%, respectively. The values of PV, FFA, TBA, TVB-N, and TMA were in the range of 8.21–10.19 meq.peroxide/kg, 1.13–1.80%, 1.03–1.19 mgMol/kg, 17.29–19.36 mgN/kg, and 2.12–2.68 mgN/kg, respectively. The results of PAH compounds in the traditional drum-smoked fish samples showed that 10 of the 16 PAH compounds detected were above the 5.0 µg/kg B(α)P maximum permissible levels stipulated by EU Regulation 1881/2006. The concentrations of Hg, Pb, Cd, and Cr in the smoked fish samples ranged from 0.001 to 0.0027 µg/g, 0.0013 to 0.0028 µg/g, 0.0014 to 0.0028 µg/g, and 0.0621to 0.0796 µg/g, respectively. The results of microbiological analysis revealed the presence of LM and SA in the traditional drum-smoked fish samples. The study concluded that traditional drum-smoked fish may constitute higher risks (chemical and microbiological) and public health concerns to consumers.     

Effect of Fish Gelatin Coating Enriched with Oregano Essential Oil on the Quality of Refrigerated Rainbow Trout Fillet

ABSTRACT

In the present study, the effect of fish gelatin coating (FG; 4%, w/v) containing oregano (Origanum vulgare L.) essential oil (OEO; 1.2% w/v) on shelf-life extension of rainbow trout (Oncorhynchus mykiss) fillet stored under refrigerated storage (4 ± 1ºC) was investigated. The control and the treated fish samples were analyzed periodically for chemical (pH; total volatile basic nitrogen, TVB-N; peroxide value, PV; thiobarbituric acid, TBA) and microbiological (total viable count, TVC; psychrotrophic count) characteristics. The FG + OEO treatment produced significantly lower (p < 0.05) TVB-N values as compared to control and FG-coated samples after Day 6 and until the end of the storage period. The TBA values remained low and were found to fluctuate during storage. Results showed that TVC exceeded 6 log CFU g^?1 on Day 8 of storage for control and FG-coated samples and Day 12 for samples coated with FG + OEO. The reduction of 1.05 and 0.83 log CFU g^?1 of psychrotrophic counts was achieved by FG + OEO coating in trout fillet (p < 0.05) as compared with control and FG-coated samples, respectively. Therefore, the incorporation of OEO into FG coating may be a promising method of maintaining the storage quality of the rainbow trout fillet stored at refrigerated temperature.     

Quality Indices of Squid (Photololigo duvaucelii) and Cuttlefish (Sepia aculeata) Stored in Ice

The objective quality indices of squid (Photololigo duvaucelii) and cuttlefish (Sepia aculeata) stored in ice were compared with the subjective counterparts. Sensory (overall quality rating, quality index method [QIM], and multisample difference test), microbiological (total viable count [TVC], psychrophilic count), chemical (trichloroacetic acid-soluble peptide [TCA-soluble peptide], trimethylamine nitrogen [TMA-N], total volatile bases nitrogen [TVB-N], ammonia content, and protein pattern), and physical analyses (expressible drip, color, and texture) were determined in both species during 16 days of iced storage. As storage time increased, TCA-soluble peptide, TVB, ammonia content, and expressible drip were increased (p < 0.05). TMA content was markedly increased after 10 and 8 days of storage in squid and cuttlefish, respectively. Both TVC and psychrophilic count increased as the storage time increased (p < 0.01). Myosin heavy chain was degraded with coincidental decrease in shear force and sensory texture during storage (p < 0.05). According to the overall rating score, shelf life of both species in ice was estimated to be 6 days. The increases in ammonia content and expressible drip were highly correlated with the decrease in overall quality rating and increase in quality index score of squid and cuttlefish (p < 0.01).     

Postmortem Biochemical and Microbiological Changes in Loricariid Catfish (Pterygoplichthys disjunctivus) Muscle During Ice Storage

ABSTRACT

Postmortem biochemical and microbiological changes in loricariid catfish (Pterygoplichthys disjunctivus) muscle were evaluated during ice storage for 20 days. Values of pH remained stable for 15 days (7.4 ± 0.2), and total volatile base-nitrogen (TVB-N) remained under acceptable limits throughout storage, with a final value of 25.2 ± 3.3 mg N/100 g muscle at Day 20. Adenosine-5´-triphosphate (ATP) and derivatives followed a postmortem degradation pattern similar to other species, with the K-value being the best freshness loss indicator. Biochemical and microbiological changes indicated that the shelf life of iced loricariid catfish muscle was 15 days under optimal (0°C) storage conditions.     

Microbial Growth Models in Gilthead Sea Bream (Sparus aurata) Stored in Ice

ABSTRACT

This study analyzes microbiological changes in whole, ungutted farmed gilthead sea bream (Sparus aurata) stored for an 18-day period in ice using traditional methods for mesophilic aerobic bacteria, psychrotrophic, Pseudomonas spp., Aeromonas spp., Shewanella putrefaciens, Enterobacteriaceae, sulphide-reducing Clostridium (Clostridia), and Photobacterium phosphoreum in muscle, skin, and gills, evaluating their seasonal differentiation. Two different statistical models were used to analyze microbiological growth. Simultaneously, physicochemical parameters such as the temperature, pH, biological oxygen demand (BOD), total dissolved solids, salinity, ammonia nitrogen, and total phosphorus content of growing waters were analyzed. The results showed that by the end of the storage time, specific spoilage bacteria (SSB) such as Pseudomonas spp., Aeromonas spp., and S. putrefaciens as H₂S-producing bacteria were dominant in sea bream harvested in temperate water in the Canary Islands. Muscle tissue had the least contamination, followed by skin and gills. The values of the analyzed seawater parameters were constant during the four seasons, except that the temperature showed a small difference between winter and summer. Seasonal effects were observed among the fish analyzed, suggesting that the lower levels of contamination detected in winter may have been due to the slight difference observed in water temperature in that season.