Evaluation of the Quality of Seer Fish

(Scomberomorus commersonii)
Stored in Dry Ice (Solid Carbon Dioxide)
M. Sasi
G. Jeyasekaran
S. A. Shanmugam
R. Jeya Shakila

ABSTRACT. The effect of dry ice (solid carbon dioxide) in preserving

seer fish (Scomberomorus commersonii) steaks showed that those packed
in 50% dry ice remained in acceptable condition up to 9 days, whereas
those packed in 20% dry ice along with 50% water ice had a longer shelf
life of 11 days. However, the seer fish steaks packed with water ice (1:1)
alone were acceptable for only 7 days. When seer fish steaks were rated
unacceptable, the TPC, psychrophilic, H2S producers, and lactics counts
reached levels of 106, 105, 105, and 104 cfu/g, respectively, but the
TMA-N and TVB-N contents were within the prescribed limits and Hx
reached a maximum of 6.66 mg%. Seer fish preserved in a combination
of dry ice and water ice was found to have a longer shelf life. [Article cop-
ies available for a fee from The Haworth Document Delivery Service:
1-800-HAWORTH. E-mail address: <docdelivery@haworthpress.com> Web-
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rights reserved.]

M. Sasi, G. Jeyasekaran, S. A. Shanmugam, and R. Jeya Shakila are all affiliated

with the Department of Fish Processing Technology, Fisheries College and Research
Institute, Tamilnadu Veterinary and Animal Sciences University, Tuticorin 628 008,
India.
Address correspondence to: G. Jeyasekaran at the above address (E-mail:
jeyasekarang@yahoo.com or ttn_jerosh99@sancharnet.in).
The authors wish to thank the Dean, Fisheries College and Research Institute, Tamil
Nadu Veterinary and Animal Sciences University, Tuticorin, India for having provided
necessary facilities to carry out this study.
Journal of Aquatic Food Product Technology, Vol. 12(2) 2003
http://www.haworthpress.com/store/product.asp?sku=J030
 2003 by The Haworth Press, Inc. All rights reserved.
10.1300/J030v12n02_06 61
62 JOURNAL OF AQUATIC FOOD PRODUCT TECHNOLOGY

KEYWORDS. Quality, seer fish, Scomberomorus commersonii, dry

ice, shelf life, water ice

INTRODUCTION

Fish undergo rapid quality changes, especially under tropical condi-

tions. Fish that have been well handled and kept at low temperatures ex-
hibit reduced bacterial growth. Water ice is commonly used for chilling
fresh fish. However, the quantity of crushed ice required for chilling
fresh fish is quite substantial (at least 1:1) and is even higher with tropi-
cal conditions. The other disadvantages of using water ice are more drip
loss, textural toughness, nutrient loss, and decreased protein extract-
ability (Putro, 1989). Hence, a need for an alternative method of chilling
fresh fish is essential to retain high quality of fresh fish.
Application of solid carbon dioxide (dry ice) is found to be effective
for preserving fish by reducing the temperature rapidly. Dry ice as a
cooling agent has certain advantages, viz., it removes three times the
quantity of heat when compared to water ice; it has a bacteriostatic ef-
fect; and it acts as an insulant enveloping the fish upon evaporation
(Putro, 1989). The dry ice is also used as a coolant in the present trend of
shipping of fresh seafoods (Schoemaker, 1990). Leblanc and Leblanc
(1992) have reported the effect of super chilling with CO2 snow on the
quality of commercially processed haddock (Melanogrammus aeglefinus)
fillets. Dry ice has recently gained popularity in India for the rapid
transportation of fresh fish by air. A significant number of fish exporters
use dry ice in combination with water ice in varying proportions for
fresh fish transportation. Since we were unable to find any scientific re-
port on the preservative effect of dry ice (solid carbon dioxide) and its
combination with water ice on the quality and shelf life extension of
fresh fish, the present study was undertaken to determine the shelf life
of a commercially important marine fish, seer fish (Scomberomorus
commersonii) preserved in dry ice and its combination with water ice.

MATERIALS AND METHODS

Whole seer fish (Scomberomorus commersonii) obtained from Tuticorin

fish landing center, India were used in this study. Dry ice (solid carbon
dioxide) manufactured by SIGGIL India Limited (Chennai, India) in 5
kg blocks was used for the experiment. Flake ice produced by a flake ice
 Sasi et al. 63

maker (ZIEGRA-EISMASCHMINEN, Germany) was used as the wa-

ter ice. Trypticase soy agar, nutrient agar, peptone iron agar, and MRS
Lactobacillus agar were obtained from Himedia Laboratories (Mumbai,
India). Analytical grade chemicals and reagents (Glaxo and S.D. Fine
Chemicals, India) were used for the biochemical analysis.
Fish (25 Nos. each weighing 2.5-3.0 kg) were beheaded, washed, and
made into steaks (without belly flaps) of size 80-120 g. The fish steaks
were then divided into three lots and each consisting of eight packs. The
first lot was packed with dry ice at the standardized level of 50%, the
second lot was packed with the standardized level of 20% dry ice along
with 50% water ice (Sasi et al., 2000) and the third lot was packed with
the recommended level (100%) of water ice (Lima dos Santos et al.,
1984), which served as control. They were designated as Packages I, II,
and III. Gloves were worn during handling of ice and fish. Care was
taken to avoid direct contact of fish and water ice with dry ice. Packages
were wrapped in polythene bags, placed in square shaped thermocole
boxes with 3 L capacity and sealed airtight with white cellophane tape
having a size of 7.5 cm. Thermocole boxes were stored at ambient tem-
perature (28 ± 2°C). Re-icing was done with the appropriate ice every
24 h to compensate for the melting loss and the melt water was drained
out. Periodically one pack containing 2 kg fish steaks were drawn from
each lot and analyzed for sensory, microbiological, and biochemical
analyses in triplicates.
Sensory characteristics and overall acceptability of fish were as-
sessed by a panel of six experienced members of the Faculty of Fish
Processing Technology on the basis of the 10-point scales described by
Huss (1988). Microbiological analysis was performed by the methods
described in APHA (1976). The total plate count, psychrophilic count,
H2S producers count, and total lactics were enumerated on trypticase
soy agar, nutrient agar, peptone iron agar, and MRS Lactobacillus agar,
respectively. The trimethylamin-nitrogen (TMA-N) and total volatile
base-nitrogen (TVB-N) contents were determined by the method of
Beatty and Gibbons (1937) using Microconway diffusion units. The
hypoxanthine (Hx) was estimated by the enzymatic method described
by Howgate (1982), using perchloric acid extracts of fish. Analysis of
Variance (ANOVA) was performed (Snedecor and Cochran, 1962) to
examine whether any significant differences existed among the differ-
ent packages of fish, with respect to the different quality characteristics
at 5% level.
64 JOURNAL OF AQUATIC FOOD PRODUCT TECHNOLOGY

RESULTS AND DISCUSSION

Sensory Evaluation
Overall sensory scores of fish stored in Packages I, II, and III are
given in Table 1. Initially, the fish had a sensory score of 9.3, which fell
below 4.5 on the 9th day of storage in Package I. However, the sensory
score of fish stored in Package II fell below 4.5 on the 11th day of stor-
age. There was a rapid reduction in sensory score of fish stored in Pack-
age III and it fell below 4.5 after 7 days. Although there was a loss in the
characteristics seaweedy odor between the 5th and 7th day of storage,
no off-flavor development was noticed in the seer fish stored in Pack-
ages I and II. However, in the seer fish stored in Package III, off-flavor
was pronounced from the 5th day onwards. The suppression of off-fla-
vor development in Packages I and II may be due to the effect of gas-
eous CO2 on the off-flavor. Bleaching and toughening were observed to
occur at a higher rate in the seer fish steaks stored in Package III, which
is mainly attributed to the fall in the fish flesh pH below a value of 6.0
(Connell, 1995). A significant difference (P < 0.05) in sensory scores
was observed only between the Packages II and III.
Microbiological Quality
Changes in total bacterial load (TPC) of fish stored in Packages I, II,
and III are presented in Table 2. The initial bacterial load of the fish was

TABLE 1. Changes in overall sensory score of seer fish (n = 3) stored in differ-

ent packages of ice

Storage Period I II III

(days)
0 9.3 ± 0.1 9.3 ± 0.1 9.3 ± 0.1
1 8.5 ± 0.3 8.8 ± 0.3 8.4 ± 0.4
3 7.8 ± 0.4 8.0 ± 0.2 6.8 ± 0.4
5 6.5 ± 0.2 7.8 ± 0.4 5.8 ± 0.3
7 5.5 ± 0.1 6.3 ± 0.2 4.6 ± 0.2
9 4.5 ± 0.2 5.8 ± 0.1 DC
11 *DC 4.3 ± 0.4
13 DC
I–Dry ice; II–Combination of dry ice and water ice; III–Water ice
* DC–Discontinued
 Sasi et al. 65

TABLE 2. Changes in total bacterial load (cfu/g) of seer fish (n = 3) stored in dif-
ferent packages of ice

Storage I II III
Period (days)
0 1.50 ± 0.04 ⫻ 104 1.50 ± 0.04 ⫻ 104 1.50 ± 0.04 ⫻ 104
1 1.03 ± 0.06 ⫻ 105 2.20 ± 0.03 ⫻ 104 3.00 ± 0.07 ⫻ 105
3 1.21 ± 0.03 ⫻ 105 5.50 ± 0.05 ⫻ 104 8.90 ± 0.02 ⫻ 106
5 1.49 ± 0.08 ⫻ 105 7.80 ± 0.02 ⫻ 104 9.80 ± 0.13 ⫻ 106
7 2.62 ± 0.07 ⫻ 105 3.92 ± 0.10 ⫻ 105 2.90 ± 0.18 ⫻ 107
9 3.72 ± 0.12 ⫻ 106 1.17 ± 0.09 ⫻ 106 DC
11 * DC 2.83 ± 0.17 ⫻ 106
13 DC
I–Dry ice; II–Combination of dry ice and water ice; III–Water ice
* DC–Discontinued

1.50 ⫻ 104 cfu/g, which increased to 3.72 ⫻ 106 cfu/g in the fish stored
in Package I on the 9th day of storage. Total bacterial load of fish stored
in Package II was 2.83 ⫻ 106 cfu/g on the 11th day of storage. However,
it increased to 2.90 ⫻ 107 cfu/g in the case of fish stored in Package III
on the 7th day of storage. The total bacterial count of seer fish in Pack-
ages I and III increased to 105 cfu/g on the first day of storage. In Pack-
age I, the bacterial load remained constant thereafter up to the 7th day,
indicating a prolonged lag period during storage. In contrast, Package II
had an initial bacterial load of 104 cfu/g and the same was maintained up
to 5 days of storage, which is an indication that the preservative effect of
CO2 snow with water ice is stronger, than the dry ice alone. However, in
Package III, there was generally an increase in the bacterial load of
about one log every two days. It has been reported that application of a
CO2 gaseous environment inhibits bacterial growth during the lag
phase, rather than during the exponential phase (Clark and Lentz,
1969), and this could be the reason for the constant bacterial load ob-
served in dry ice (CO2 snow) stored seer fish (Packages I and II). CO2 is
reported to inhibit the growth of active spoilage bacteria, such as
Alteromonas and Pseudomonas (Barnett et al., 1971), which might have
lowered the bacterial counts in Packages I and II. A significant differ-
ence (P < 0.05) was observed in the total bacterial load of the seer fish
stored in Packages II and III, and I and III.
Total psychrophiles in the fish stored in Packages I, II, and III were
found to be 103 cfu/g, 104 cfu/g, and 105 cfu/g, respectively, on the 1st
66 JOURNAL OF AQUATIC FOOD PRODUCT TECHNOLOGY

TABLE 3. Changes in total psychrophiles (cfu/g) of seer fish (n = 3) stored in

different packages of ice

Storage Period I II III

(days)
0 < 1.00 ± 0.01 ⫻ 101 < 1.00 ± 0.01 ⫻ 101 < 1.00 ± 0.01 ⫻ 101
1 7.00 ± 0.07 ⫻ 103 1.50 ± 0.05 ⫻ 104 2.80 ± 0.09 ⫻ 105
3 2.21 ± 0.08 ⫻ 104 2.50 ± 0.03 ⫻ 104 1.02 ± 0.14 ⫻ 106
5 5.50 ± 0.07 ⫻ 104 4.00 ± 0.05 ⫻ 104 4.30 ± 0.10 ⫻ 106
7 7.50 ± 0.11 ⫻ 104 1.18 ± 0.04 ⫻ 105 8.40 ± 0.12 ⫻ 106
9 1.60 ± 0.10 ⫻ 105 3.92 ± 0.09 ⫻ 105 DC
11 * DC 6.00 ± 0.13 ⫻ 105
13 DC
I–Dry ice; II–Combination of dry ice and water ice; III–Water ice
* DC–Discontinued

day of storage (Table 3). A log increase in the count was observed on the
3rd day and thereafter, it remained constant up to 7 days in the fish stored
in Packages I and III. In the case of fish stored in Package II, there was al-
most no change in the psychrophilic count from the 1st day to the 5th day.
At the end of the storage period, the fish stored in Packages I and II had a
load of 105 cfu/g, while the load was 106 cfu/g in the fish stored in Pack-
age III. The higher psychrophilic bacterial count encountered in Pack-
age III was mainly due to the favorable effect of low refrigeration
temperature brought about by the water ice for growth of bacteria. The
lower psychrophilic bacterial count observed in Packages I and II was
mainly because of the effect of dry ice (CO2 snow), which creates a gas-
eous environment and a much lower temperature than the normal water
ice. It was reported that Gram-negative psychrophilic spoilage bacteria
were more susceptible to CO2 (Gill and Tan, 1980), which may also be a
reason for the reduction in the psychrophilic bacterial count in Packages
I and II. A significant difference (P < 0.05) in psychrophilic counts be-
tween the seer fish stored in Packages I and III, and II and III was ob-
served.
The fish samples stored in Packages I, II, and III were found to con-
tain H2S producers at the level of 102 cfu/g (Table 4) on the 1st day of
storage. A constant increase in the load by one log every two days was
observed in the fish stored in Package III. On the other hand, H2S pro-
ducers in the fish stored in Packages I and II exhibited a slower growth.
 Sasi et al. 67

TABLE 4. Changes in H2S producers (cfu/g) of seer fish (n = 3) stored in differ-

ent packages of ice

Storage Period I II III

(days)
0 < 1.00 ± 0.01 ⫻ 101 < 1.00 ± 0.01 ⫻ 101 < 1.00 ± 0.01 ⫻ 101
1 2.20 ± 0.04 ⫻ 102 5.00 ± 0.03 ⫻ 102 6.50 ± 0.05 ⫻ 102
3 4.00 ± 0.07 ⫻ 103 1.13 ± 0.06 ⫻ 103 7.70 ± 0.09 ⫻ 103
5 1.20 ± 0.09 ⫻ 104 1.65 ± 0.08 ⫻ 104 4.89 ± 0.12 ⫻ 104
7 6.00 ± 0.13 ⫻ 104 6.20 ± 0.10 ⫻ 104 9.94 ± 0.15 ⫻ 105
9 7.30 ± 0.18 ⫻ 105 2.31 ± 0.13 ⫻ 105 DC
11 * DC 8.26 ± 0.14 ⫻ 105
13 DC
I–Dry ice; II–Combination of dry ice and water ice; III–Water ice
* DC–Discontinued

The H2S-producing bacteria were found to be lower than the total

bacterial and psychrophilic bacterial counts. A similar observation was
recorded in cod fish stored at 2°C by Jensen et al. (1980) and Huss
(1988). The H2S-producing bacterial count increased more or less uni-
formly throughout the storage period in all the packages of seer fish
from 101 to 105 cfu/g. Earlier investigations also showed that CO2 has
an inhibitory effect on H2S-producing bacteria (Jensen et al., 1980).
The increase of the H2S-producing bacterial count in seer fish stored in
different packages of ice was significant (P < 0.05).
Changes in total lactics in fish stored in Packages I, II, and III are pre-
sented in Table 5. Fish stored in Packages I and II had a higher load of
lactics (103 cfu/g) than the fish stored in Package III (102 cfu/g) on the
3rd day of storage. A gradual rise in lactics was observed in the fish
stored in Package III, whereas, the load was constant in fish stored in
Package II from the 3rd day to the 7th day and thereafter, more or less a
log increase was noticed. However, in Package I, the lactics count
reached a level of 104 cfu/g on the 5th day, which was higher than the
counts observed in Packages II and III. Dry ice provided a favorable en-
vironment for the lactics, and this was reflected in the counts, which in-
creased further from 103 to 105 cfu/g in Package I. Whereas, Package II
provided only a minimally favorable environment for the lactics, as the
counts only reached 104 cfu/g during the storage period. On the other
hand, in the fish stored in Package III, the lactic counts increased by
only a log after the 3rd day, which then very gradually increased to
68 JOURNAL OF AQUATIC FOOD PRODUCT TECHNOLOGY

TABLE 5. Changes in total lactics (cfu/g) of seer fish (n = 3) stored in different

packages of ice

Storage Period I II III

(days)
0 < 1.00 ± 0.01 ⫻ 101 < 1.00 ± 0.01 ⫻ 101 < 1.00 ± 0.01 ⫻ 101
1 < 1.00 ± 0.01 ⫻ 101 < 1.00 ± 0.01 ⫻ 101 < 1.00 ± 0.01 ⫻ 101
3 1.70 ± 0.03 ⫻ 103 2.60 ± 0.03 ⫻ 103 9.00 ± 0.02 ⫻ 102
5 1.40 ± 0.05 ⫻ 104 1.50 ± 0.04 ⫻ 103 1.54 ± 0.04 ⫻ 103
7 5.50 ± 0.06 ⫻ 104 4.70 ± 0.05 ⫻ 103 2.28 ± 0.06 ⫻ 104
9 1.86 ± 0.10 ⫻ 105 1.11 ± 0.08 ⫻ 104 DC
11 * DC 6.30 ± 0.09 ⫻ 104
13 DC
I–Dry ice; II–Combination of dry ice and water ice; III–Water ice
* DC–Discontinued

104 cfu/g. Jay (1987) has reported that most of the lactics can grow in
the pH range of 4.0 to 4.5. In the present study, since the pH of the seer
fish stored in different packages of ice varied from 6.1 to 6.3, the multi-
plication of lactics was not significant. However, the lactic counts ob-
served in the seer fish stored in different packages of ice were not
significant (P < 0.05).

Biochemical Quality

Changes in trimethylamine-nitrogen (TMA-N) content of fish stored

in Packages I, II, and III are given in Table 6. The initial TMA-N con-
tent of fish was 1.78 mg%, which increased to 5.33 mg% in the fish
stored in Package I on the 9th day of storage, while it was only 4.27
mg% in the fish stored in Package II. But, the fish stored in Package III
had a higher TMA-N level of 7.46 mg% on the 7th day of storage. How-
ever, the TMA-N contents observed in all of the packages of fish did not
exceed the acceptable limit of 10-15 mg% (Connell, 1995) even at the
time of rejection. It has been reported that in marine fish, the rate of in-
crease in TMA varies considerably varies from species to species (Amu
and Disney, 1973; Huss, 1988). Though it has been suggested that CO2
has an inhibitory effect on TMAO reducing bacteria, there was a report
that in a MAP fish system, 20% CO2 has inhibited the TMA-N produc-
tion for 14 days, but 40% CO2, only for 3 days (Brown et al., 1980). The
results of the present study also show that TMA-N cannot be univer-
 Sasi et al. 69

sally used as an index of fish quality. The changes in TMA-N contents

of seer fish stored only in Packages II and III were significant (P < 0.05).
The seer fish had an initial TVB-N level of 3.91 mg% (Table 7). The
rise in TVB-N was more rapid in the fish stored in Package III, with
Package II having the lowest value on the 9th day. The TVB-N content
of seer fish steaks stored in different packages of ice was well within the
limit of acceptability of 30 mg% (Connell, 1995) until the end of stor-
age. The TVB-N values of seer fish stored in Package I ranged from
3.91 to 25.25 mg%. Oberlender et al. (1983) have reported that the

TABLE 6. Changes in trimethylamine-nitrogen (TMA-N) content (mg%) of seer

fish (n = 3) stored in different packages of ice

Storage Period (days) I II III

0 1.78 ± 0.04 1.78 ± 0.04 1.78 ± 0.04
1 2.49 ± 0.13 1.78 ± 0.14 2.13 ± 0.07
3 2.49 ± 0.08 2.13 ± 0.08 3.56 ± 0.08
5 4.27 ± 0.07 2.85 ± 0.08 5.33 ± 0.04
7 5.33 ± 0.14 3.56 ± 0.09 7.46 ± 0.13
9 5.33 ± 0.05 4.27 ± 0.03 DC
11 *DC 4.62 ± 0.14
13 DC
I–Dry ice; II–Combination of dry ice and water ice; III–Water ice
* DC–Discontinued

TABLE 7. Changes in total volatile base-nitrogen (TVB-N) content (mg%) of

seer fish (n = 3) stored in different packages of ice

Storage Period (days) I II III

0 3.91 ± 0.04 3.91 ± 0.14 3.91 ± 0.14
1 8.89 ± 0.11 7.82 ± 0.07 10.31 ± 0.05
3 10.67 ± 0.08 11.38 ± 0.04 13.51 ± 0.03
5 13.87 ± 0.07 13.16 ± 0.06 19.56 ± 0.06
7 15.65 ± 0.14 16.01 ± 0.12 24.53 ± 0.15
9 25.25 ± 0.17 20.62 ± 0.14 DC
11 *DC 24.18 ± 0.05
13 DC
I–Dry ice; II–Combination of dry ice and water ice; III–Water ice
* DC–Discontinued
70 JOURNAL OF AQUATIC FOOD PRODUCT TECHNOLOGY

TVB-N contents of MAP-stored swordfish steaks were about 11.14

mg% of the 8th day. The lower level of TVB-N contents in the seer fish
stored in Packages I and II when compared to that stored in Package III
was mainly because of the presence of CO2 (dry ice) that inhibited the
microorganisms responsible for the formation of TVB-N (Oberlender et
al.,1983). Significant difference (P < 0.05) was noticed in the TVB-N
contents of seer fish stored in Packages I and II with that stored in Pack-
age III.
Initial hypoxanthine (Hx) content of the fish was 0.24 mg% (Table 8).
The rise in Hx content was rapid (9.28 mg%) in the fish stored in Pack-
age III. On the other hand, the fish stored in Package II had a low level
of hypoxanthine (4.88 mg%), even when they were at the threshold of
spoilage. However, the fish stored in Package I had an intermediate
level of hypoxanthine (6.66 mg%) at the end of storage. The lower Hx
values recorded in fish stored in Package II coincided well with the sen-
sory and most of the microbiological and biochemical characteristics.
Brown et al. (1980) have also reported that Hx values of MAP-stored
rock fish showed wide variation during storage, but, remained unaf-
fected by the modified atmospheric (CO2) treatment. In the present
study also, no significant different (P < 0.05) in Hx was observed
among the different packages of ice.
The results showed that the seer fish steaks stored in a combination of
dry ice and water ice had a longer shelf life of 11 days, followed by the
seer fish steaks stored in dry ice with 9 days, when compared to their
storage in water ice with 7 days. As the quality of seer fish steaks stored

TABLE 8. Changes in hypoxanthine (Hx) content (mg%) of seer fish (n = 3)

stored in different packages of ice

Storage Period (days) I II III

0 0.24 ± 0.07 0.24 ± 0.07 0.24 ± 0.07
1 0.19 ± 0.13 0.31 ± 0.15 1.07 ± 0.05
3 2.74 ± 0.08 0.79 ± 0.07 3.14 ± 0.07
5 4.40 ± 0.07 2.49 ± 0.04 6.90 ± 0.13
7 5.59 ± 0.07 3.21 ± 0.08 9.28 ± 0.12
9 6.66 ± 0.09 4.52 ± 0.11 DC
11 *DC 4.88 ± 0.07
13 DC
I–Dry ice; II–Combination of dry ice and water ice; III–Water ice
* DC–Discontinued
 Sasi et al. 71

in dry ice and its combination with water ice was found to be exception-
ally better during the initial period of storage than in simple water ice,
such fish will certainly fetch a higher price when exported in fresh con-
dition and, thereby, increase the revenue for the seafood industry. The
use of dry ice alone was found to be cost prohibitive, as higher concen-
tration was required for effective chilling, while its combination with
water ice at lower concentration was found to be effective, economical,
and best suited for the short-term preservation and transportation of
high valued fishes like seer fish.

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IBH Publishing Co., Calcutta. pp. 339-380.

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