EFFECTS OF VIETNAMESE TAMARIND FISH SAUCE ENRICHED WITH IRON AND ZINC ON GREEN MUSSEL QUALITY
Abstract and keywords
Abstract (English):
Marinating is a traditional technology to improve the shelf - life and quality of products. Enriched Viet- namese tamarind fish sauce was prepared and used to marinate green mussels. Subjects of this research were: green mussels (C1), marinated green mussels (C2), and marinated green mussels packaged in modified atmosphere (M1, M2, and M3). A percentage of O :CO :N was 5:50:45 for M1, 5:70:25 for M2, and 5:90:5 for M3. Microbiological, hemical, and sensory qualities of the samples were analyzed during storage at 4°C for 30 days. The results indicate that glycogen, iron, and zinc contents as well as pH values decreased in all the cases during storage. In addition, TVB-N and TBARS values as well as mesophilic and lactic acid bacteria increased with time in all the samples. The number of psychrophilic bacteria in marinated samples was small or not detected at all. Escherichia coli, Staphylo- coccus aureus, Salmonella spp. and Vibrio cholera were below standard values. Based on sensory acceptability, the shelf-life of C1, C2, M1, M2, and M3 samples was 12, 18, 24, 24, and 27 days, respectively.

Keywords:
Vietnamese tamarind fish sauce, iron, zinc, marinated green mussel, modified atmosphere packaging (MAP)
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Green mussel (Perna viridis L.) is a common ma- rine animal in the Asia-Pacific region [1]. This is a large bivalve with smooth, elongated  shell  typical  of  seve- ral mytilids. Flesh  of  the  green  mussel  is  con- sidered  an  excellent   source   of   selenium,   cal- cium,       iron,       magnesium,       phosphorous       and

vitamins   (A,    B1,    B2,   B6,    B12   and    C)   [2,    3]. In addition, mussel fat is rich in polyunsaturated fat- ty acids (PUFA,  37–48% of total  fatty acids) mainly

ω–3 PUFA [4], which is biologically important and can decrease the risk of cardiovascular disease [5]. Howe- ver, this filter feeder animal has a high risk of microbio- logical contamination that lead to a short shelf-life, poor quality, and  economic  losses.  In  addition,  only  well– cooked green mussels are microbiologically safe.

Marinating is a seasoning process mainly using or- ganic acid containing liquids [6]. Liquid marinades ge-

 

nerally include sugar, spices, oil, and acid such as vine- gar, fruit juice or wine. Marinating improves tenderness, juiciness, taste and aroma, as well as improves the shelf- life of red meat, poultry, seafood, and vegetables [7–10].

Vietnamese tamarind fish sauce is popular in its country of origin. The main ingredients are tamarind pulp, fish sauce and sugar, while some fresh garlic and chili are used to enhance taste. The Vietnamese con- sume this sauce with various foods, such as seafood, fried food, dried food. It is even used as a condiment in various kinds of food products. Ready-to-cook pro- ducts are expected to gain popularity in South-East Asia as well. Mineral deficiencies, including iron and zinc, remain problematic in developing countries. Thus, the enrichment of food products with minerals can be an effective way to solve this problem. However,  accor- ding to a market survey, green mussels marinated with enriched tamarind fish sauce are not commercially avai-

 

 

Copyright © 2019, Tien et al. This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), allowing third parties to copy and redistribute the material in any medium or format and to remix, transform, and build upon the material for any purpose, even commercially, provided the original work is properly cited and states its license.

 

 

 

lable, especially when packed in a modified atmosphere. Modified atmosphere packaging (MAP) is a technology that prolongs the shelf-life of food products by retard- ing microbial growth and biochemical reactions, often with an elevated carbon dioxide level [11]. Moreover, in MAP oxygen may inhibit the growth of anaerobic bacte- ria and accumulate toxins from Clostridium botulinum, type E [12]. However, botulinum toxins can form at be- low 2% oxygen [13, 14].

Therefore, the aim of this study was to determine the shelf-life of green mussels marinated with tamarind fish sauce enriched with iron and zinc. The mussels were packed in modified atmosphere and stored at 4°C.

 

STUDY OBJECTS AND METHODS

2

 
Materials. Tamarind pulp without seeds, fish sauce, white sugar, finger chili, garlic, and the fresh green mussels  (40–50  individuals/kg)  were  purchased  from a supermarket located in Songkhla province. Expe- riments were carried out at the Food Technology De- partment, the Agro-Industry Faculty of  the Prince of Songkla University, Thailand. Iron sodium ethylenedi- aminotetraacetate (FeNaEDTA·3(H O)) and zinc sulfate

(ZnSO ·7(H O)) were taken from Sigma–Aldrich Com-

 

foam agent were added. 5 ml from 50 ml of the distillate were mixed with 5 ml of 0.02 M 2-thiobarbituric acid in 90% glacial acetic acid (TBA reagent). The mixture was placed in a vial that was capped and heated in a water bath for 35 min. Afterwards, the vial was cooled down and the sample was measured at 532 nm using a spec- trophotometer (UV/VIS T180, PG Instrument Ltd., UK). The results are expressed in mg malondialdehyde/kg sample.

Glycogen content. Glycogen content was deter- mined based on the method of [16]. 50 mg of chopped marinated green mussels were boiled for 20 min with 400 µl of 33% KOH, cooled down, and 700 µl of 96% ethanol was added. The mixture was placed in an ice bath for 2 hours to reach complete precipitation. There- after, the mixture was centrifuged at 7,500 g for 20 min using a microlitre centrifuge (Z-233-M2, Hermle– Germany) and the supernatant was collected. The su- pernatant was mixed with 1 ml of distilled water and sonicated for 5 min using Crest Ultrasonic (575 DAE, Malaysia). Then 100 µl of the solution and 2 ml of an- throne reagent were kept at 90°C for 20 min in dark- ness. The reagent consisted of 38 ml of concentrated

sulphuric acid with 15 ml of distilled water and 0.075 g

 

4            2

 

pany (Steinheim, Germany).

Marinated green mussel preparation. The green mussels were washed during 2 hours with tap water, then blanched in boiling water at 100°C for 30 s to fa- cilitate meat removal. A ratio of sample to water was 1:2  w/w.  Afterwards,  the  meat  of   green   mussels was separated from the shells. We prepared tam- arind fish sauce contained  42%  of  tamarind  slur- ry, 37% of sugar, and 21% of fish sauce.   236 mg of

FeNaEDTA·3(H O)   and   88.5 mg   of   ZnSO ·7(H O)

 

of anthrone. The solution was measured at 620 nm using

a spectrophotometer. Glycogen concentration was calcu- lated from a standard curve for glycogen from Sigma– Aldrich Company (Steinheim, Germany).

Total volatile base nitrogen (TVBN). Total vola- tile base (TVBN) contents in the green mussels were determined using Conway micro-diffusion method de- scribed in [17]. 2 g of a sample were mixed with 8 ml of 4% tricloroacetic acid (TCA), then homogenized with

a high-speed homogenizer for 1 min. The supernatant

 

2                                                                                      4            2

 

were added into 30 g of the sauce, which was 36 mg of Fe and 20 mg of Zn, respectively. The green mus- sel meat was soaked in the enriched sauce at the ra- tio of 1:1.  The  marinated  meat  was  packaged  in 15 х 30 cm Havel Vacuum bags, b.v. (Europac Co., Ltd)

 

was filtered by Whatman No. 41 filter paper (Whatman

2         3

 
International, Ltd., Maidstone, UK). 1 ml of the sample was placed in the outer ring, while 1% boric acid con- taining the Conway indicator was pipetted into the inner ring. 1 ml of saturated K CO  was added to the sam-

 

2

 
with  O

transmission  rate  46.6  cm3m2   day–1    at  38°C,

 

ple to initiate the reaction after mixing. Thereafter, the

 

1 atm. We tested five samples of green mussels: meat without marinade solution packed under normal air (C1), marinated meat without gases (C2), and meat packed in the modified atmosphere (M1, M2, and M3). A percent-

 

mixture was incubated at room temperature for 3 hours.

Afterwards, the inner ring solution was titrated using 0.02N HCl until the green color turned to pink similar to a blank sample. TVB-N content was calculated as fol-

 

age of O :CO :N

 

was 5:50:45 for M1, 5:70:25 for M2,

 

lows, mg/100 g:

 

2           2      2

 

and 5:90:5 for M3. All samples were stored at 4°C for

30 days. Microbiological, chemical and sensory analyses were performed every 3 days.

pH. The marinated green mussels were blended with sterilized distilled water at the ratio of 1:5 and allowed to stand for 2 min. Then pH was measured by using a pH meter (Mettler 350, Singapore).

Thiobarbituric acid reactive substances (TBARS). The thiobarbituric acid reactive substances (TBARS) were determined by the method described in [15]. 10 g of chopped samples were homogenized with 50 ml of distilled water for 2 min and then transferred to distil- lation tubes. 47.5 ml of distilled water was additional- ly infused into the distillation tube to avoid losing the

sample. 2.5 ml of 4 N HCl and a few drops of an anti-

 

(mg/100 g) ,

where: N is normality of HCl in titrant; A is titration vo- lume for actual sample; B is titration volume for blank; V is total initial volume of sample with TCA.

Iron and zinc content. Samples of 0.5 g in mass were digested in 5 ml of concentrated nitric acid with 2 ml of 30% hydrogen peroxide under pressure in a closed vessel before heating in a microwave oven un- til the samples were digested. Afterwards,  the  sam- ples were cooled down and filtered by Whatman filter paper No. 1 with a pore size of 110 nm (Whatman In-

ternational, Ltd., Maidstone, UK)). Each sample solu-

 

 

 

tion was then transferred into a 25 ml volumetric flask and the volume was made up with distilled water [18]. The extract samples were used to determine iron and zinc by Inductively Couple Plasma Mass Spectrometry (ICP-MS) (Perkin-Elmer SCIEX, Shelton, CT, USA). The RF power was 1,350 W, the plasma gas flow rate was 15 l/min, the carrier gas flow was 0.94 l/min, and make-up gas flow was 0.15 l/min. The sample take rate was 100 µl/min and the sample depth was 6.0 mm.

Microbiological quality. 25 g of the marinated green mussels were blended with 225 ml of 0.1% ste- rilized peptone water. From the 10–1 dilution, dilutions to 10–2 and 10–3  were prepared with 0.1% sterilized peptone water. Mesophilic and psychrophilic bacteria, Esche- richia coli, Salmonella spp, Vibrio cholera, and Staphy- lococcus aureus were  determined  using  the  method in [19]. Lactic acid bacteria were determined using de Man Rogosa and Shape agar by pour plate method, with incubation in anaerobic conditions at 35°C for 48 hours before colony count [20].

Sensory evaluation. Marinated samples were baked in a casserole pan (House worth HW-1707S, China) at 180°C for 10 min until the core temperature of the meat sample reached 80 ± 2°C. Then the samples were served to fifty untrained panelists of post-graduate students and technicians from the Department of Food Technology, Prince of Songkla University. The panelists were asked to evaluate the likability scores for appearance, aroma, texture, taste and overall on a nine-point hedonic scale.

Statistical analyses.  All experiments  were  run in triplicates. The data were subjected to Analysis of Vari- ance (ANOVA) and the differences between means were assessed for significance by Duncan’s Multiple Range Test [21]. Data analysis was performed with the SPSS package (SPSS 6.0 for Windows, SPSS Inc., Chicago, IL, USA).

 

RESULTS AND DISCUSSION

We found that the sauce had pH 3.31, total titratable acidity 32.13%, and water activity 0.87. The viscosity of this enriched sauce was 840 cP, with non-Newtonian be- havior. The colour coordinates L*, a*, and b* were 26.44,

15.8 and 33.7, respectively.

pH. pH levels of un-treated and treated green mus- sels are shown in Fig. 1. In general, pH with all treat- ments decreased with storage time. The initial pH of un-treated green mussels (C1) was 6.77 and decreased to

6.21 at 15 days of storage (p < 0.05). The marinated sam- ples had significantly lower pH than the control group due to acidity of the sauce (pH 3.31) (p < 0.05). The pH of marinated green mussels without gases (C2) was

4.48 on day 1 and declined to 4.07 by day 21 of storage (p < 0.05). The initial pH of marinated green mussel (M1, M2, and M3) was about 4.51–4.64 and dropped to 4.24, 4.2 and 3.95, respectively, at 30 days of storage (p < 0.05). The decrease in pH of meat during chilled storage is normally caused by conversion of glycogen to lactic acid and other volatile acids, as a result of mus- cle degradation [22]. Cao et al. also reported that the spoilage of mollusk shellfish was partly by fermentation

indicated by decreasing pH [23]. Moreover, pH was low-

 

 

,

 

 

Fig. 1. Dependence of pH values of green mussels on storage time. C1 = control; C2 = marinated with no gases; M1, M2, and M3 = marinated at the O :СO :N  percentage of 5:50:45;

2          2      2

5:70:25; and 5:90:5, respectively.

 

2

2

due to the dissolution of CO  that  produced  carbonic acid [24] and reduced production of alkaline compounds in the marinated samples [24, 25]. Higher concentrations of CO (M3 > M2 > M1) gave lower pH levels in our ex- periments, although without significant differences. The pH value in our experiments was mainly due to the aci- dic marinade sauce with tamarind pulp.

TBARS. Fig. 2  demonstrates  the  TBARS  levels in green mussels during storage for the various treat- ments. TBARS is indicator of the malondialdehyde con- tent, which is a secondary lipid oxidation product from polyunsaturated fatty acids [26] causing off-flavors in meat products [27]. The initial TBARS of the control (C1) was 2.62 mg MDA/kg, while the marinated sam- ples (C2, M1, M2, M3) had 0.99, 0.81, 0.92, 0.88 mg

,

,

Подпись: ,MDA/kg, respectively. It should be noted that the mari- nated green mussels had lower TBARS values than the control samples, apparently due to osmosis during ma- rinating conveying TBARS out from the mussels. Also the bacterial count was higher for C1 than for the other samples, which could also play a role in producing TBARS [28]. The lower TBARS values of the marina-

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Fig. 2. Dependence of TBARS values of green mussel

on storage time. C1 = control; C2 = marinated with no gases; M1, M2, and M3 = marinated at the O :СO :N  percentage

 

2          2      2

 

er with MAP using gases than with other treatments,

 

of 5:50:45; 5:70:25; and 5:90:5, respectively.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

,

2,400

 

1,800

 

1,200

 

600

 

0

,

Fig. 3. Dependence of TVB – N values of green mussels on storage time. C1 = control; C2 = marinated with no gases; M1, M2, and M3 = marinated at the O :СO :N  percentage

 

 

Fig. 4. Dependence of glycogen content of green mussels

on storage time. C1 = control; C2 = marinated with no gases; M1, M2, and M3 = marinated at the O :СO :N  percentage of

 

2          2      2

 

2          2      2

 

of 5:50:45; 5:70:25; and 5:90:5, respectively.

 

5:50:45; 5:70:25; and 5:90:5, respectively.

 

 

 

ted green mussels were consistent throughout the sto- rage, and sulfur compounds from garlic in the marinade may have played a role as antioxidants [29]. However, TBARS in all cases increased during storage because of high content of polyunsaturated fatty acids in mussel fat, as mentioned in the introduction. Moreover, MAP packaging reduced TBARS may be due to O2-elimina-

 

and M1 samples were 27, 24, and 25 days, respectively. In addition, [36] noticed that for seafood TVB-N indi- cated the freshness of raw materials and acceptability for human consumption. They pointed out that TVB-N increased with storage time as a result of quality dete- rioration. However, the initial TVB-N of the marinated products may not indicate the quality, considering that

 

2

 
tion by enriched CO

proportion [22]. Masniyom et al.

 

fish sauce and garlic used as ingredients cause erroneous

 

reported that seabass flesh in MAP did not suffer from

increased lipid oxidation [30]. The TBARS  level  of 5–8 mg/kg malonaldehyde is a standard criterion of the poor quality of oily food [31, 32]. TBARS in the control (C1) reached 8.03 mg MDA/kg on day 12 of storage, while C2 had 7.85 mg MDA/kg on day 18. M1, M2, and M3 had TBARS levels below the standard on day 27, namely 7.59, 6.89, and 6.82 mg MDA/kg, respectively. The marinated green mussels packed at the percentage

of O2:CO2:N2 equaled 5:90:5 gave the least TBARS level at the end of storage (p > 0.05).

Total volatile base nitrogen (TVB-N). Changes in the TVB-N value of the treated and un-treated green mussels during storage are shown in Fig. 3. The TVB-N increased with storage time. The initial TVB-N of green mussels (C1) was 6.44 mg/100 g and increased gradually to 10.22 mg/100 g during storage. The treated samples C2, M1, M2, and M3 had initial TVB-N values 20.6,

 

results.

Glycogen content. During storage glycogen content decreased gradually in all samples, but declined rapidly in C2 samples (p < 0.05), as shown in Fig. 4. The ini- tial glycogen content in the control sample (C1) was sig- nificantly lower than that in the marinated samples (C2, M1, M2, and M3). Glycogen is a polysaccharide includ- ing glucose units in a branched structure. Extraction and hydrolysis were used to obtain simple sugars in the gly- cogen content determination. As mentioned earlier, the marinade solution had 37% of sugar, so it positively con- tributed to the glycogen content. Jermyn [37] mentioned

 

Table 1. Iron and zinc content in green mussel samples during storage

 

Minerals                   Samples  Initially                 End of storage

 

18.97, 19.53 and 18.94 mg/100 g, respectively. By the end of the storage they reached 30.1, 33.33, 33.67, and

29.69 mg/100 g, respectively (p < 0.05). TVB-N is com- posed of  volatile amines,  typically  dimethylamine, trimethylamine and ammonia [22], that commonly de- termine the spoilage of seafood [33]. The marinated samples had higher TVB-N values than the control even on day 1, due to volatile amines in the fish sauce [34]. In addition, the marinade contained garlic with sulfur compounds that could also react with boric acid, con- tributing to TVB-N content [29]. The increasing trend

of TVB-N with time in this study agrees with the find-

 

Iron content,

mg/kg

 

 

 

 

 

Zinc content, mg/kg

 

C1            71.44 ± 0.77a           67.61 ± 0.67b

C2            599.62 ± 12.60a      371.47 ± 2.33b M1               389.67 ± 1.14a            381.34 ± 1.99b M2 399.2 ± 1.98a               390.00 ± 1.41b M3                481.93 ± 1.36a        392.36 ± 0.44b C1  11.09 ± 0.40a               8.95 ± 0.13b C2      382.19 ± 8.22a            249.00 ± 1.41b M1 267.20 ± 1.56a            257.74 ± 2.51b M2             280.35 ± 0.64a            266.17 ± 1.17b M3 331.00 ± 1.98a            271.11 ± 2.40b

 

ings of [35].  Fishery products are acceptable for human

 

Note: a–b   Means within rows with different superscripts are signifi-

cantly different (p < 0.05). C1 = control; C2 = marinated with no

 

consumption when TVB-N does not exceed 30 mg/100 g

 

gases; M1, M2, and M3 = marinated at the O :СO :N

 

percentage of

 

,

Подпись: ,

,

Подпись: ,of sample [34]. Based on this, the shelf-lives of M3, M2,

 

 

5:50:45; 5:70:25; and 5:90:5, respectively

 

2         2      2

 

 

Table 2. Microbiological levels in marinated and non-marinated green mussel kept under different conditions during the storage at 4°C for 30 days.

 

 

E.coli, MPN/g     Psychrophile,

CFU/g

Подпись: E.coli, MPN/g     Psychrophile,
CFU/g

Mesophile, CFU/g

Подпись: Mesophile, CFU/gBac-

 

 

Treat-

 

Storage time, days

 

 

terial

type

ment

1

3

6

9

12

15

18

21

24

27

30

 

C1

9.3 × 103

7.0 × 104

2.5 × 105

6.3 × 105

8.6 × 105

1.2 × 106

 

 

 

 

 

 

C2

2.8 × 103

3.7 × 103

8.9 × 104

9.8× 104

1.04 × 105

5.3 × 105

7.1 × 105

1.13 × 106

 

 

 

 

M1

< 100

3.6 × 103

6.5 × 103

7.9 × 103

8.6 × 103

1.30 × 104

1.63 × 104

1.90 × 104

2.45 × 104

9.0 × 104

1.42 × 106

 

M2

< 100

2.5 × 103

5.2 × 103

5.7 × 103

6.3 × 103

9.2× 102

1.05 × 104

1.47 × 104

2.10 × 104

2.6 × 104

1.12 × 106

 

M3

< 100

< 100

< 100

3.0 × 103

5.2 × 103

7.4 × 103

9.6 × 103

1.10 × 104

1.80 × 104

2.48 × 104

9.8 × 105

 

C1

8.5 × 103

1.56 × 104

2.0 × 104

2.13 × 104

2.32 × 104

5.0 × 104

 

 

 

 

 

 

C2

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

 

 

 

 

M1

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

 

M2

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

 

M3

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

< 10

 

C1

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

 

 

 

 

 

C2

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

 

 

 

M1

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

M2

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

<  3.0

 

M3

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

C1

ND

ND

ND

ND

ND

ND

ND

ND

ND

ND

ND

 

C2

ND

ND

ND

ND

ND

ND

ND

ND

ND

ND

ND

 

M1

ND

ND

ND

ND

ND

ND

ND

ND

ND

ND

ND

 

M2

ND

ND

ND

ND

ND

ND

ND

ND

ND

ND

ND

 

M3

ND

ND

ND

ND

ND

ND

ND

ND

ND

ND

ND

 

C1

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

 

 

 

 

 

C2

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

 

 

 

M1

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

M2

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

M3

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

C1

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

 

 

 

 

 

C2

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

 

 

 

M1

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

M2

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

M3

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

< 3.0

 

C1

9.9 × 103

1.5 × 104

1.76 × 104

2.1 × 104

2.6 × 104

3.5 × 104

 

 

 

 

 

 

C2

6.0 × 103

1.02 × 104

1.66 × 104

1.81× 104

2.41 × 104

4.8 × 104

7.2 × 104

1.14 × 105

 

 

 

 

M1

8.0 × 103

9.5 × 103

1.43 × 104

1.72 × 104

2.10 × 104

2.5 × 104

4.6 × 104

6.2 × 104

8.5 × 104

1.12 × 105

2.12 × 105

 

M2

6.0 × 103

7.6 × 103

9.4 × 103

1.02 × 104

1.71 × 104

2.38 × 104

2.5 × 104

3.5 × 104

4.1 × 104

5.6 × 104

1.36 × 105

 

M3

2.5 × 103

3.4 × 103

5.4 × 103

8.3 × 103

1.34 × 104

1.87 × 104

2.25 × 104

3.2 × 104

3.5 × 104

4.2 × 104

6.2 × 104

 

Lactic acid bac-   V.cholerae,

Подпись: Lactic acid bac-   V.cholerae,

teria , CFU/g

Подпись: teria , CFU/g

MPN/g

Подпись: MPN/gNote: C1 = control, C2 = normal marinated, M1 = marinated at 5%O :50%CO :45%N , M2 = 5%O :70%CO :25%N

 

 

M3 = 5%O :90%CO :5%N

 

2                   2               2

 

2                   2                2,

 

2                   2             2

 

 

 

S.aureus,

MPN/g

Подпись: S.aureus,
MPN/g

Salmonella

spp., CFU/g

Подпись: Salmonella
spp., CFU/g
that in the determination of carbohydrates by anthrone method, other simple sugars in the system interfered. He pointed out that using glycogen content for glycolysis determination, which should decrease with storage time, might not give good results for high sugar products. The glycogen content decreased with storage time due to me- tabolism by glycolysis pathway in some bacteria [22]. The glycogen content of the control sample (C1) dropped to 61.42 mg/kg by day 12 of storage, while the marinat- ed without MAP sample (C2) had 614.01 mg/kg after 18 days (p < 0.05). In the marinated samples packed with gases (M1, M2, and M3) the glycogen contents were 857.33; 1,005.68; and 1,293.17 mg/kg. The marinade with sugar strongly increased glycogen levels in marinated products. However, in C1the glycogen content decreased from 94.75 to 61.42 mg/kg during 12 days. The rapid de- crease in glycogen levels of C2 may due to faster bacte- rial growth than with the treatments that included MAP, and the garlic from marinade solution seems to acceler- ate glycogen loss relative to the control (C1) perhaps by

 

 

sulfur compounds and prebiotics in garlic that support lactic acid bacterial growth [22].

Iron and zinc content. Iron and zinc contents of green mussels with  various  treatments  are  present- ed in Table 1. On day 1, the iron content in C1 was

71.44 mg/kg, while that in the marinated samples C2, M1, M2, and M3 was 599.62; 389.67; 399.2; and 481.93 mg/kg, respectively. The initial zinc contents in C1, C2, M1, M2, and M3 were 11.09; 382.19; 267.2; 280.35; and 331 mg/kg, respectively. The iron and zinc contents may have fluc- tuated due to uncontrolled factors, such as uneven mar- inade absorption, the amount of marinade drip from the surface of green mussels during sample preparation for testing, or the small weight (1.0 g) of samples tested. If the samples are not uniform, then consistent results may not be obtained. The iron and zinc content in the mari- nated samples was higher than that in the control sample due to the absorption of iron and zinc from the mari- nade. Generally, the iron and zinc contents decreased by

the end of the shelf-life to 67.61; 371.47; 381.34; 390.00;

 

 

Table 3. Sensory acceptability score of green mussel kept under different condition during storage at 4°C for 30 days

 

 

Storage time,

 

Treatments                                                                               Attributes

 

day

Appearance

Flavor

Colour

Texture

Taste

Overall

1

C1

6.63 ± 0.72a

6.40 ± 0.62b

6.90 ± 0.99a

7.07 ± 0.74ab

6.57 ± 0.68a

6.63 ± 0.73b

 

C2

6.90 ± 0.76a

7.10 ± 0.84a

6.90 ± 0.80a

7.30 ± 0.75a

7.03 ± 0.61a

7.10 ± 0.84a

 

M1

6.93 ± 0.98a

6.57 ± 0.63b

6.73 ± 0.69a

6.73 ± 0.52bc

6.77 ± 0.68a

6.60 ± 0.56b

 

M2

6.83 ± 0.65a

6.80 ± 0.71ab

6.53 ± 0.57a

6.33 ± 0.48c

6.77 ± 0.73a

6.60 ± 0.67b

 

M3

6.90 ± 0.61a

6.80 ± 0.81ab

6.90 ± 0.96a

6.83 ± 0.70a bc

6.83 ± 0.70a

6.80 ± 0.48ab

3

C1

6.50 ± 0.73a

6.27 ± 0.98b

6.47 ± 0.78a

6.97 ± 0.85a

6.40 ± 0.93b

6.23 ± 0.94c

 

C2

6.83 ± 0.91a

6.87 ± 0.94a

6.73 ± 0.83a

6.80 ± 0.71a

6.83 ± 0.83ab

6.80 ± 0.76ab

 

M1

6.60 ± 0.81a

6.40 ± 0.67b

6.57 ± 0.50a

6.63 ± 0.93ab

6.50 ± 0.86ab

6.53 ± 0.78bc

 

M2

6.70 ± 0.79a

6.90 ± 0.84a

6.60 ± 0.67a

6.27 ± 0.52b

6.40 ± 0.56b

6.43 ± 0.50bc

 

M3

6.83 ± 0.79a

6.67 ± 0.76ab

6.83 ± 0.75a

6.77 ± 0.63a

6.93 ± 0.78a

6.97 ± 0.96a

6

C1

6.23 ± 0.63b

6.13 ± 0.43c

6.43 ± 0.63ab

6.77 ± 0.82a

6.70 ± 0.53abc

6.13 ± 0.68c

 

C2

6.67 ± 0.71a

6.80 ± 0.92a

6.70 ± 0.88a

6.57 ± 0.50abc

6.80 ± 0.76a

6.77 ± 0.90a

 

M1

6.60 ± 0.72a

6.30 ± 0.53bc

6.27 ± 0.52b

6.33 ± 0.61bc

6.43 ± 0.63bc

6.57 ± 0.57ab

 

M2

6.60 ± 0.56a

6.53 ± 0.63ab

6.40 ± 0.50b

6.20 ± 0.76c

6.40 ± 0.62c

6.27 ± 0.52bc

 

M3

6.83 ± 0.70a

6.63 ± 0.67ab

6.60 ± 0.67ab

6.70 ± 0.70ab

6.77 ± 0.68ab

6.80 ± 0.71a

9

C1

6.17 ± 0.53b

6.03 ± 0.41c

6.13 ± 0.68b

6.50 ± 0.51ab

6.03 ± 0.61b

6.07 ± 0.69b

 

C2

6.57 ± 0.68a

6.70 ± 0.79a

7.00 ± 0.79a

6.20 ± 0.61bc

6.57 ± 0.90a

6.63 ± 0.76a

 

M1

6.47 ± 0.51ab

6.27 ± 0.69bc

6.13 ± 0.63b

6.23 ± 0.73bc

6.30 ± 0.65ab

6.37 ± 0.56bc

 

M2

6.53 ± 0.63a

6.43 ± 0.50ab

6.30 ± 0.47b

6.13 ± 0.57c

6.30 ± 0.60ab

6.17 ± 0.59b

 

M3

6.47 ± 0.73ab

6.53 ± 0.73ab

6.50 ± 0.78b

6.60 ± 0.67a

6.63 ± 0.61a

6.70 ± 0.60a

12

C1

6.10 ± 0.55ab

5.73 ± 0.58b

6.03 ± 0.41c

6.17 ± 0.53b

5.87 ± 0.57c

6.00 ± 0.45b

 

C2

6.03 ± 0.76b

6.27 ± 0.45a

6.57 ± 0.68a

6.13 ± 0.63b

6.20 ± 0.48bc

6.13 ± 0.43b

 

M1

6.37 ± 0.49ab

6.17 ± 0.38a

6.07 ± 0.52c

6.17 ± 0.59b

6.17 ± 0.53b

6.27 ± 0.58b

 

M2

6.47 ± 0.57a

6.40 ± 0.72a

6.23 ± 0.50c

6.07 ± 0.58b

6.23 ± 0.63ab

6.10 ± 0.66b

 

M3

6.30 ± 0.53ab

6.37 ± 0.56a

6.43 ± 0.82ab

6.50 ± 0.63a

6.50 ± 0.63a

6.57 ± 0.63a

15

C2

5.90 ± 0.66a

6.17 ± 0.91a

6.40 ± 0.77a

6.10 ± 0.76a

6.13 ± 0.76a

6.10 ± 0.61b

 

M1

6.23 ± 0.57a

6.07 ± 0.78a

6.00 ± 0.91b

6.10 ± 0.40a

6.07 ± 0.37a

6.13 ± 0.63ab

 

M2

6.20 ± 0.61a

6.27 ± 0.45a

6.17 ± 0.38ab

6.03 ± 0.49a

6.20 ± 0.66a

6.03 ± 0.67a

 

M3

6.27 ± 0.52a

6.23 ± 0.68a

6.33 ± 0.61a

6.43 ± 0.68a

6.27 ± 0.45a

6.37 ± 0.76a

18

C2

5.83 ± 0.79a

5.93 ± 0.78a

6.27 ± 0.78a

6.03 ± 0.67a

5.97 ± 0.76a

5.90 ± 0.66b

 

M1

6.13 ± 0.57a

6.03 ± 0.56a

5.87 ± 0.455b

6.07±0.45a

6.00 ± 0.79a

6.07 ± 0.64ab

 

M2

6.17 ± 0.75a

6.13 ± 0.63a

6.10 ± 0.66ab

6.00 ± 0.69a

6.13 ± 0.43a

6.00 ± 0.74ab

 

M3

6.17 ± 0.38a

6.17 ± 0.53a

6.27 ± 0.58a

6.30 ± 0.47a

6.18 ± 0.46a

6.30 ± 0.47a

21

M1

6.07 ± 0.52a

5.90 ± 0.61a

5.70 ± 0.75b

6.00 ± 0.53a

5.93 ± 0.52a

6.03 ± 0.72a

 

M2

6.10 ± 0.76a

6.07 ± 1.11a

6.07 ± 0.64a

5.90 ± 0.55b

6.07 ± 0.58a

5.97 ± 0.61a

 

M3

6.10 ± 0.55a

6.13 ± 0.63a

6.17 ± 0.59a

6.23 ± 0.43a

6.10 ± 0.31a

6.23 ± 0.50a

24

M1

6.03 ± 0.72a

5.90 ± 0.80a

5.70 ± 0.84a

5.97 ± 0.67a

5.80 ± 0.71a

5.97 ± 0.76a

 

M2

6.00 ± 0.53a

6.03 ± 0.76a

5.93 ± 0.64a

5.90 ± 0.61a

6.00 ± 0.37a

5.87 ± 0.63a

 

M3

6.03 ± 0.72a

6.03 ± 0.56a

6.10 ± 1.09a

6.13 ± 0.51a

6.00 ± 0.45a

6.13 ± 0.82a

27

M3

5.93 ± 0.37

5.97 ± 0.72

5.93 ± 0.58

5.90 ± 0.55

6.00 ± 0.26

6.07 ± 0.37

Note: Mean ± SD from 50 panalists; a–f Means within a column with different superscripts are significantly different (p < 0.05). C1 = control;

C2 = marinated with no gases; M1, M2, and M3 = marinated at the O :СO :N  percentage of 5:50:45; 5:70:25; and 5:90:5, respectively

2         2      2

 

 

and 392.36 mg/kg for C1, C2, M1, M2, and M3, respec- tively. The zinc contents similarly were 8.95; 249.00; 257.74; 266.17; and 271.11 mg/kg for the same samples. The decrease in iron and zinc contents during storage may be due to the degradation of proteins by browning reaction, which may easily interact with iron and zinc.

El-Din et al. [38] mentioned that  when  iron  and zinc minerals were added into dairy products, such as milk, the iron was bound with amino acids into casein micelles. Zinc associated with colloidal calcium phos- phate in the casein micelles. Therefore, the contents of

available iron and zinc may be reduced. In addition, it

 

was noted that the drip increased with storage time due to high salt content (~ 5.5% NaCl) and acidity of the marinade, leaching iron and zinc out from the mussels. Baygar et al. [39] observed that water content decreased during marinating.

Microbiological quality. Mesophilic and lactic acid bacteria were found in all treated samples throughout the experiment. However, psychrophilic bacteria were found only in the control sample (C1), while Escherich- ia coli, Staphylococcus aureus, Salmonella spp., and Vibrio cholera were very low or not detected, as shown

in Table 2. Psychrophilic bacteria in the marinated sam-

 

 

 

ples (C2, M1, M2, and M3) were present in the amount of no more than 10 CFU/g. This indicates that the hur-

 

various  treatments  during  storage  are  summarized  in Table 3. The scores from panelists became worse with

 

w

 
dle effect of the sauce (a

0.87, pH 3.31, TA 32.13%, and

 

storage time (p < 0.05). The threshold score for rejection

 

2

 
salt 5.26%) strongly controlled psychrophilic bacterial growth. During storage, mesophilic and lactic acid bac- teria increased with time (p < 0.05). Not surprisingly that the bacterial count in the control sample (C1) was the highest, followed by the marinated samples (C2), (M1), (M2) and (M3) in this order. ICMSF [40] recom- mended 106CFU/g as the standard limit of total viable count in freshwater fish and seafood. By this thresh- old the shelf-life of the control was 12 days, while the marinated green mussel (C2) had 18 days. With higher CO  contents lower bacterial counts were found. This

was chosen to be 6, and only aroma showed significant differences between un-treated and treated green mussels during storage (Table 3). While all the sensory attributes with any treatment degraded towards the end of shelf-life, the scores remained on an acceptable level. Unexpectedly, the marinated mussels did not score better than the con- trol group. This may be due to too soft texture, and unusu- al aroma and taste from the sauce that was highly acidic and contained iron and zinc. This altered the good natural aroma and texture of the mussels. This type of sauces is served as dips in Vietnam, not as marinades.

 

suggests  that  MAP  can  prolong  the  shelf-life  when

 

2

 
CO   inhibits microbial growth. Farber [25] also stat-
CONCLUSION

 

2

 
ed  that  CO

becomed  more  antibacterial  as  its  con-

 

TVB-N, TBARS, TVC, and lactic acid bacteria in-

 

2

 
centration increased. CO

can decarboxylate enzymes

 

creased in all samples during storage, while pH, glyco-

 

and inhibit the metabolic activity of microflora, there-

by extending the lag phase and reducing logarithimic phase growth of spoilage bacteria [25, 41, 12]. Goulas

 

gen, iron, and zinc contents decreased. The shelf-life of marinated green mussel was extended from 12 to 18, 24, or 27 days based on microbiological quality, TBARS and

 

et al. [42] reported that when mussels (Mytilus gallo-

 

TVB-N value, by use of CO

 

rich MAP packaging. For

 

provincialis) were kept in 80%CO :20%N

 

atmosphere,                                                                                            2

 

2                   2                                                 marinated green mussels, high concentration of CO

 

(90%)

 

the total viable count of bacteria was retarded compared

to other tested conditions. Masniyom et al. [43] also re-

 

2

appears to be both feasible and beneficial to shelf-life.

 

2

 
ported that MAP with 80–100% of CO

effectively ex-

 
CONFLICT OF INTEREST

 

tended the shelf-life of green mussels (Perna viridis).

 

2

 
Our current study indicates that 90% CO

atmosphere

 

The authors declare no conflict of interest.

 

is the most effective to prolong the shelf-life of mar- inated  green  mussels.  Therefore,  combined  acidity,

 
ACKNOWLEDGMENTS

 

2

 
salt and atmospheric CO microbial growth.

were effective in retarding

 

This work was supported by the Higher Education Research Promotion and the Thailand’s Education Hub

 

Sensory evaluation. Generally,  sensory  evalu- ation is applied  in  the  estimation  of  seafood  quali- ty and correlates well with the microbiological and chemical analyses [44].  The  sensorial  scores  from  fif- ty  non-trained  panelists  for  green  mussels  with  the

 

for Southern Region of ASEAN Countries Project Office

of the Higher Education Commission.

We would like to thank the team of the Research and Development Office (RDO) of the Prince of Songkla University, Thailand for the assistance with English.

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