International Journal of Advanced Research in Engineering and Technology

(IJARET)
Volume 6, Issue 10, Oct 2015, pp. 51-61, Article ID: IJARET_06_10_010
Available online at
http://www.iaeme.com/IJARET/issues.asp?JType=IJARET&VType=6&IType=10
ISSN Print: 0976-6480 and ISSN Online: 0976-6499
IAEME Publication
___________________________________________________________________________

DEHYDROGENASES AND ASCORBIC ACID

PROFILE WITH MICROBIAL ANALYSIS
BETWEEN TWO GREEN MUSSEL (PERNA
VIRIDIS) POPULATIONS
S.P. Indumathi1, R. Karthik2&3, Angelin C Pushpam2&3,
M.C. Vanitha2&3 and K. Ramalingam1*
1
Department of Zoology, Government Arts College,
Nandanam, Chennai- 600035, Tamil Nadu, India
2
Department of Marine Biotechnology, AMET University (U/S 3 of UGC Act 1956),
Kanathur, Chennai 603112, Tamil Nadu, India
3
Centre for Marine Bioprospecting, AMET University (U/S 3 of UGC Act 1956),
Kanathur, Chennai 603112, Tamil Nadu, India
ABSTRACT
The marine mollusks are essential since most of them are edible and used
as food. It has received distinct attention and become a natural resource of
economic importance. Perna species, like other bivalves are filter feeders are
mostly cultured in coastal and estuarine areas. The coastal area is much
polluted by the human impacts especially by industrial and sewage
discharges. The organisms grow under such pollution prone zone are much
affected and there is an variation in the metabolic, enzymatic and other
biochemical components. The oxido reductases of the mitochondrial system
such as lactic dehydrogenase (LDH), Malate dehydrogenase (MDH) and
Succinate dehydrogenase (SDH) plays a key role in most of the metabolic
pathways. The presence of ascorbic acid acts as defence mechanism during
the stress condition. The essential dehydrogenases and the antioxidant along
with the microbial content were investigated. In the present study the Perna
viridis was selected from two sites, one from the relatively unpolluted zone
about one km away from the shore and the other group from the polluted zone
located along the shore line for comparative study on the dehydrogenases,
ascorbic acid and microbial content in the relation to the varied
environmental characteristics. The reduction in the enzymes and ascorbic acid
profile and increase of microbial population in the polluted zone reveals that
the organism is under stress and is not viable for consuming.

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S.P. Indumathi, R. Karthik, Angelin C Pushpam, M.C. Vanitha and K. Ramalingam

Keywords: Mussel Population, Antioxidant, Dehydrogenases, Pollution,

Shoreline.
Cite this Article: S.P. Indumathi, R. Karthik, Angelin C Pushpam, M.C.
Vanitha and K. Ramalingam. Dehydrogenases and Ascorbic Acid Profile with
Microbial Analysis between Two Green Mussel (Perna Viridis) Populations.
International Journal of Advanced Research in Engineering and Technology,
6(10), 2015, pp. 51-61.
http://www.iaeme.com/IJARET/issues.asp?JType=IJARET&VType=6&IType=10

1. INTRODUCTION
The marine resources of India being enormous, molluscs contribute to a great extend
to the food resource. In India, the average annual landings of total bivalve during
1995-2000 was estimated as 1, 50, 103 tonnes of which mussels represented 11, 316
tonnes, although the potential yield was estimated to be 2, 080 tonnes. The various
molluscan species include oysters, clams and mussels. The last mentioned species i.e.,
mussels are an important source of fishery along the coastal areas. The bivalve
molluscs occupy the inter-tidal region, the back waters and also the region of waters
off the coastline (Dhandapani, 2001). The demand for mussels of good quality far
exceeds the supply. Coastal areas with domestic and industrial pollution load could
reduce their population in spite of their reproductive capacity, breeding potential and
growth (Nelson et al., 1988). The living organisms found in poor environmental
condition or in polluted waters are considered to be under stress. Such stress not only
may have a direct bearing in reducing the overall biodiversity but could affect the
growth, density and distribution of a single species which is biologically and
economically important which leads to the dominance of tolerant varieties, thus
affecting the catch both qualitatively and quantitatively. The variations in biochemical
constituents do exist with the progression of maturity (Nagabhushnam and Mane,
1978). During the last three decades the coastal regions have become a sink and an
inlet for various pollutants originated by the anthropological activities of human
population (Swarnalatha, 1998). The various categories of pollutants that reach the
coastal habitats where the molluscs inhabit include the heavy metals, pesticides,
domestic sewage and industrial effluents etc. Enzymes serve as indicators of tissue
and molecular malfunction. Based on their function enzymes are categorized into
oxidative, hydrolytic and transaminating enzymes. Any stress subjected to the
organisms would alter the enzymatic level since the cells or tissues undergo direct
damage. Such changes have been reported in the activities of lactate dehydrogenase
(LDH), succinate dehydrogenase (SDH), glutamate oxaloacetate transaminase (GOT),
glutamate pyruvate transaminase (GPT), acid phosphatase (ACP), and alkaline
phosphatase (ALP) in animals subjected to drug poisoning (Payne and Payne, 1985).
This information indicates the functional status of various organs and the impact of
the strain in stressed conditions. The results obtained indicates how the enymatic
profiles and their consequent changes result in the manifestation of pathological
changes of tissues and organs concerned, and morbidity of the animal.
The present study was proposed to delineate the impact of pollution factor on this
species, revealed their occurrence in two different zones viz., near the coast and off
the coast (one kilometre away from the coast), i.e., species occurring in pollution free
and species occurring in pollution prone zone. The enzyme concentrations, ascorbic
acid content along with the microbial analysis were viewed to obtain the information
on the species growth, development and food value.

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Dehydrogenases and Ascorbic Acid Profile with Microbial Analysis Between Two Green
Mussel (Perna Viridis) Populations

2. MATERIALS AND METHODS

2.1. Collection and Identification of Green Mussels
The live specimens Green mussel Perna viridis were handpicked from submerged
rocky areas of Royapuram coastal zone. The specimens collected one kilometre away
from the shoreline (Station 1) and collected from the shoreline (Station 2). They were
considered as the control and as the polluted group respectively. Animals measuring
about 90-100 mm in length were selected for the study irrespective of sex. 12 mussels
were taken to carry out the following analysis. The specimens collected were
identified by its morphological features. (Buddo et al. 2003).

2.2. Haemolymph Collection and Tissue Separation

The haemolymph from Perna viridis was collected following the method mentioned
by (Santarem and Figueras, 1995). A plastic rod was carefully inserted between the
shell valves and the mussel was taken out from the medium. The needle was inserted
with a 5ml sterile plastic syringe, into the posterior adductor muscle to withdraw the
colourless haemolymph. For tissue analysis shells were split open and dissected to
obtain the tissues in digestive gland, adductor muscle, foot and gills. The tissues from
three animals weigh mg the same, were collected for the analysis.

2.3. Determination of Tissue and Haemolymph Lactate Dehydrogenase

The lactate dehydrogenase activity was determined following the procedure adopted
by King (1965). 100mg wet tissue of digestive gland, adductor muscle, foot and gill
regions of Perna viridis from control viz., unpolluted and polluted groups were
weighed and homogenized separately in 100 ml ice cold double distilled water
centrifuged at 3000rpm for 15 mins. 0.1ml of the buffered substrate were taken in two
different test tubes. 0.1ml of centrifuged supernatant of the tissue extract and to the
control 1ml of distilled water was added. 0.2 ml of NADH solution was added to both
the test tubes incubated exactly for 15 mins at 37C. At the end of this period 1.0 ml
of 2, 4- dinitrophenol hydrazine reagent was added to each tube incubated at 37c for
15mins and 10ml of 0.4N sodium hydroxide was added to each tube. The intensity of
the colour was measured at 440nm, in Baush and Lomb Spectronic21
Spectrophotometer exactly 60 seconds after the addition of the alkali. The amount of
LDH activity was calculated from the OD of the standard and expressed as
MIU/min/mg protein.
0.1ml of the haemolymph and 1ml of ice cold double distilled water were taken
and centrifuged at 3000 rpm for 15mins. The procedure for the tissue extract was
applied for the estimation of haemolymph LDH.

2.4. Determination of Tissue and Haemolymph Malate Dehydrogenase

MDH activity was determined following the procedure adopted by Aquiline and
Farmsworth (1960). 100mg of wet tissue was weighed and homogenised using 5 ml of
ice cold double distilled water centrifuged at 3000 rpm for 15mins and supernatant
thus obtained was used for measurement of enzyme activity. 2.5 ml of Sorenson
buffer, 0.2ml of the tissue extract and 0.2ml of co-enzyme solutions were mixed in a
test tube and allowed to stand at room temperature for 15-20mins. Freshly prepared
substrate 0.2ml and mixed well and the decrease in the adsorption of the reaction
mixture, at the gap of one min interval for 5 min were recorded at 340 nm in Baush

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S.P. Indumathi, R. Karthik, Angelin C Pushpam, M.C. Vanitha and K. Ramalingam

and Lomb Spectronic21 Spectrophotometer. The enzyme activity is expressed in

MIU/min/mg protein.
To 0.5ml of the haemolymph 0.5ml of ice cold water was added and centrifuged at
3000rpm for 15 min. the supernatant thus obtained was used for the measurement of
enzyme activity. The MDH activity was estimated in haemolymph extract following
the procedure applied for tissue MDH activity and the result was determined in
MIU/min/mg protein.

2.5. Determination of Tissue and haemolymph Succinate dehydrogenase

The method used to estimate SDH activity is that of Nachlas et al. (1960). 100mg of
wet tissue was weighed and homogenised using 10 ml of 0.25M sucrose solution and
centrifuged at 3000rpm for 15 min and the supernatants thus obtained was used for
the measurement of enzyme activity. The reaction mixture contained 0.4ml of 40mM
sodium succinate. 0.5ml of 100mM sodium phosphate buffer pH 7.4, 0.5ml of 2mM
INT. The volume was made upto 2 ml by adding 0.6ml of double distilled water.
0.5ml of the extract was added and then incubated for 30 min at 37C. 5ml of glacial
acetic acid was added and the colour was extracted in 5ml of toluene by keeping in
the refrigerator overnight at 10C. The colour developed was measured at 495 nm
against toluene blank in a Baush and Lomb Spectronic21 Spectrophotometer. Enzyme
activity of the tissue was expressed in MIU/min/mg protein.
To 0.5ml of the haemolymph 0.5ml of ice cold water was added and centrifuged at
3000rpm for 15 min. the supernatant thus obtained was used for the measurement of
enzyme activity. The MDH activity was estimated in haemolymph extract following
the procedure applied for tissue MDH activity and the result was determined in
MIU/min/mg protein.

2.6. Determination of Ascorbic acid (Vitamin c) Antioxidant

Ascorbic acid content was determined calorimetrically following the method of
Sadasivam and Manikam (1996). 10-100g standard dehydroascorbic solution was
pipette out into a series of tubes. Similarly different aliquots (0.1ml-2ml) of
brominated sample extracts were pipetted out. Volume in each test tube was made
upto 3 ml by adding distilled water. One ml of DNPH reagent was added followed by
1-2 drops of thiourea to each tube. A blank was set as above with water in place of
ascorbic acid solution. The content of the tube was mixed thoroughly and incubated at
37C for 3 hrs. After incubation the orange red osazone crystals formed was dissolved
by adding 7ml of 80% sulphuric acid. The absorbance was measured at 540 nm. A
graph of ascorbic acid contents in the sample was calculated. The results were
expressed in g/100 mg wet tissue.
1ml of the haemolymph was taken for analysis by the above procedure.

3. MICROBIAL ANALYSIS
Bacterial count in the surface water and raw meat of Perna viridis control viz.,
unpolluted and polluted groups were made according to the standard methods (APHA,
1998).

3.1. Microbial analysis in water samples

Samples were collected at polluted and non-polluted sites in sterile borosil bottles
leaving ample air space. Samples were transported to the laboratory within hours for
analysis. The sterilized medium was poured in sterile petri plates and was allowed to
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Dehydrogenases and Ascorbic Acid Profile with Microbial Analysis Between Two Green
Mussel (Perna Viridis) Populations

solidify. 1ml of diluted water samples was spread over the medium and plates were
kept inverted and incubated for 24 h at 37C. The numbers of colonies were counted
after the incubation period (cfu/ml).

3.2. Microbial analysis of the tissue homogenate for bacteria

The peptone water was prepared by dissolving 5 g of peptone in 500 ml of water
followed by the addition of 2.5 gms of sodium chloride, the pH was adjusted to
7.20.2. the solution was sterilization at 15 lbs pressure (121C) for 15 minutes.
About 5 gms of mussel meat from control viz., unpolluted and polluted groups were
aseptically removed and transferred to a sterile mortor. To this, 45 ml of peptone
water was added. The meat was ground well to form tissue homogenate. This formed
1:10 dilution which was then serially diluted decimally (10-1 to 10-6). 0.1ml from each
dilution was pipetted aseptically into culture dishes. Uninoculated plates serve as
control. The plates were inverted and incubated at 37C and bacterial count was done
after the incubation period (cfu/g).

3.3. Statistical analysis

In the present investigation tissues and haemolymph of Perna viridis from viz.,
unpolluted and polluted groups were taken as factors. The statistical analysis was
performed with Analysis of variance (ANOVA), F-test and Turkey test (t-test) for
comparison of data (Zar, 1974 and Kanji, 1999).

4. RESULTS
4.1. Lactate dehydrogenase activity (LDH)
The lactate dehydrogenase activity in the various tissues and haemolymph of Perna
viridis control viz., unpolluted group versus polluted are given in Table 1. The level of
lactate dehydrogenase activity showed a significantly lowered activity in the digestive
gland, adductor muscle, foot, gill and haemolymph of the control viz., unpolluted
group when compared to polluted group. In the polluted group, the level of lactate
dehydrogenase activity in the respective tissues and haemolymph showed a
significantly higher level. The differences within the groups were found to be
statistically significant (P<0.01).
Table 1 Lactate Dehydrogenase (LDH) activity in the various tissues (MIU/min/mg protein)
and haemolymph (MIU/min/mg protein) of Perna viridis. Control/unpolluted Vs polluted
t value

P value

SD
0.0014

188197.00

620.742c

0.0011

387413.00

0.0012

964.062e

0.0015

380282.00

175.938a

0.0008

460.556a

0.0028

242551.00

Haemolymph

322.564b

0.0028

470.465b

0.0054

85498.00

P
0.01
P
0.01
P
0.01
P
0.01
P
0.01

P value

P < 0.01

SD
0.0034

Polluted
Mean
910.633d

325.175c

0.0015

656.789e

Gill

Digestive
gland
Adducator
Gland
Foot

Control
Mean
630.929d

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<
<
<
<
<

P < 0.01

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S.P. Indumathi, R. Karthik, Angelin C Pushpam, M.C. Vanitha and K. Ramalingam

Each value represents mean SD of six observations.

P value 0.01denotes significant at 1% level.
Different alphabets within groups denotes significant at 5% level.

5. MALATE DEHYDROGENASE ACTIVITY (MDH)

The level of malate dehydrogenase activity in the various tissues and haemolymph of
Perna viridis control viz., unpolluted group versus polluted group are given in Table
2. The malate dehydrogenase activity showed a significantly higher activity in the
digestive gland, adductor muscle, foot, gill except for the haemolymph of control viz.,
unpolluted group when compared to the polluted group. In the polluted group the
levels of malate dehydrogenase activity showed a significantly higher level. The
differences within the groups were found to be statistically significant (P<0.01).
Table 2 Malate Dehydrogenase (MDH) activity in the various tissues (MIU/min/mg protein)
and haemolymph (MIU/min/mg protein) of Perna viridis. Control/unpolluted Vs polluted
Control
Mean
0.006523d

SD
0.000015

Polluted
Mean
SD
b
0.001954
0.000032

0.008500e

0.000022

0.001578a

0.000022

541.56

0.002960b

0.000013

0.002859c

0.000007

17.27

Gill

0.006373c

0.000029

0.006248a

0.000033

7.03

Haemolymph

0.001367a

0.000035

0.003402d

0.000033

103.06

P value

P < 0.01

Digestive
gland
Adducator
Gland
Foot

t value

P value

320.39

P
0.01
P
0.01
P
0.01
P
0.01
P
0.01

<
<
<
<
<

P < 0.01

Each value represents mean SD of six observations.

P value 0.01denotes significant at 1% level.
Different alphabets within groups denotes significant at 5% level.

6. SUCCINATE DEHYDROGENASE ACTIVITY (SDH)

The level of succinate dehydrogenase activity in the various tissues and haemolymph
of Perna viridis control viz., unpolluted group versus polluted group are given in
Table 3. The succinate dehydrogenase activity showed a significantly higher level in
the digestive gland, adductor muscle, gill and haemolymph except in the foot of
control viz., unpolluted group when compared to the polluted group. In the polluted
group, the respective tissues and haemolymph except for the foot showed a
significantly lowered activity. The differences between the tissues within the groups
were found to be statistically significant (P<0.01).

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Dehydrogenases and Ascorbic Acid Profile with Microbial Analysis Between Two Green
Mussel (Perna Viridis) Populations
Table 3 Succinate Dehydrogenase (SDH) activity in the various tissues (MIU/min/mg
protein) and haemolymph (MIU/min/mg protein) of Perna viridis. Control/unpolluted Vs
polluted
Control
Mean
SD
e
Digestive gland
9929.667 0.02981
Adducator Gland 1873.548d 0.1451
Foot
1108.643c 0.2280
Gill
496.2967a 0.0484
Haemolymph
837.6067b 0.0841
P value
P < 0.01

Polluted
Mean
SD
e
6242.455 0.1184
1809.525c 0.1618
1990.535d 0.1532
219.4533a 0.1585
237.6517b 0.2161
P < 0.01

t value
28156.10
721.49
7864.00
4092.72
6337.99

P value
P < 0.01
P < 0.01
P < 0.01
P < 0.01
P < 0.01

Each value represents mean SD of six observations.

P value 0.01denotes significant at 1% level.
Different alphabets within groups denotes significant at 5% level.

7. ASCORBIC ACID (VITAMINS-C)

The level of ascorbic acid in the various tissues and haemolymph of Perna viridis
control viz.,unpolluted group versus polluted group are presented in Table 4. The
ascorbic acid level showed a significantly higher level in the digestive gland, adductor
muscle, foot, gill and haemolymph of the control viz.,unpolluted group when
compared to the polluted group. In the polluted group the levels of ascorbic acid in the
different tissues and haemolymph showed a significantly lower level. The difference
within the groups were found to be statistically significant (P < 0.01).
Table 4 Ascorbic acid (Vitamin-C) Levels in the various tissues (g/100 mg wet tissue) and
haemolymph (g/ml) of Perna viridis. Control/unpolluted Vs polluted
Control
Mean
SD
c
537.2631
0.0044
e
816.6478
0.0026

Polluted
Mean
SD
e
300.5534
0.0032
d
296.6303
0.0048

106369.00
234988.00

P < 0.01
P < 0.01

623.3778d

0.0037

265.1247c

0.0034

173361.00

P < 0.01

Gill

97.7147a

0.0042

27.2626a

0.0031

32999.80

P < 0.01

Haemolymph

127.3240b

0.0029

28.2307b

0.0023

65016.70

P < 0.01

P value

P < 0.01

Digestive gland
Adducator
Gland
Foot

t value

P value

P < 0.01

Each value represents mean SD of six observations.

P value 0.01denotes significant at 1% level.
Different alphabets within groups denotes significant at 5% level.

8. MICROBIAL ANALYSIS
The microbial analysis revealed the presence of bacteria in both the water samples.
Comparison revealed higher colonies in the polluted waters in all the serial dilutions
Table 5a. The bacterial count in the raw meat of Perna viridis ranged from 11 x 10-2;
8 x 10-3 and 2 x 10-4 in the control group. Similarly in the polluted group the range
was found to be
16 x 10-2; 11 x 10-3 and 7 x 10-3 respectively. There were no

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S.P. Indumathi, R. Karthik, Angelin C Pushpam, M.C. Vanitha and K. Ramalingam

colonies in X10-5 and X10-6 in both the water samples. These results along with
infective doses of bacteria that could cause food poisoning to human being is
presented in Table 5b. The comparison suggests that the bacterial contamination was
found to be within the permissible limits.
Table 5a Variation in microbial cell number of the surface seawater from the non-polluted
(control) station-1 and polluted site (station-2) (Cfu/ml)
Serial Dilution
X10-1
X10-2
X10-3
X10-4
X10-5
X10-6

Station-1, Unpolluted Water

(control)
Exceeds 300 colonies
Exceeds 300 colonies
43 x 10-3
28 x 10-4
Nil
Nil

Station-2 Polluted Water

Exceeds 300 colonies
Exceeds 300 colonies
55 x 10-3
44 x 10-4
10 x 10-5
3 x 10-6

Table 5b Bacterial count in the raw meat of Perna viridis. Results expressed in Cfu/gm
Serial Dilution
X10-1
X10-2
X10-3
X10-4
X10-5
X10-6
Infective dose for human consumption

Raw meat from control viz.

unpolluted group
Exceeds 300 colonies
11 x 10-2
8 x 10-3
2 x 10-4
Nil
Nil
X10-6 to X10-10

Raw meat from

polluted group
Exceeds 300 colonies
16 x 10-2
11 x 10-3
7 x 10-4
Nil
Nil

9. DISCUSSION
The increase in mussel cultivation has brought an increasing awareness. Compared to
the environmental stressors, the man-made stressors remain unique in that they
impose severe stress over the already conditioned and genetically adapted organisms
like molluscs. Hence the quantitative estimation of the degree of the above super
imposed stress of man-made origin seemed to be pertinent to diagnose the health of
the animals and also to find ways and means to do better ecological resource
management of the above forms.
Oxidoreductases of the mitochondrial system include lactic dehydrogenase
(LDH), malate dehydrogenase (MDH) and Succinate dehydrogenase (SDH).
Considerable work on LDH, MDH and SDH activity of animals intoxicated by
pesticides and other biocides have been carried out to implicate the toxic stress on
them (Koundiya and Ramamurthi, 1978; Bhagyalakshmi et al., 1984; Shubashini et
al., 1985). The enzyme lactate dehydrogenase represents a key point in the flow of
carbon through glycolysis and citric acid cycle and the physiological status of an
animal.(De Zwaan and Van Marrewijk, 1973; Gade and Zeba, 1973; Holwerda and
De Zwaan. 1973; Hammen, 1975). In the present study, the results on LDH activity
showed an overall increase in all the tissues of the polluted group over that of the
control viz., unpolluted group. Previous studies that have been made on LDH of
molluscs are very much meagre with regard to natural contaminants or toxicants of
man made origin. The increased LDH activity in both hepatopancreas and body
muscle was noticed in the prawn Caridina rajadhari exposed to lethal and sublethal
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Dehydrogenases and Ascorbic Acid Profile with Microbial Analysis Between Two Green
Mussel (Perna Viridis) Populations

holothurian biotoxin (Kumar et al., 1991). Reddy and Rao (1986) reported similar
increase in LDH activity in the tissues of hepatopancreas and body muscle of Penaeus
indicus treated with phosphomidon. In contrast, Reddy and Rao (1991) observed
decreased LDH activity in the midgut gland and muscle tissue of the Penaeid prawn,
Metapenaeous monoceros exposed to methyl parathion. Malate dehydrogenase
catalyses the reversible oxidation of oxaloacetate to L-malate. Enzymes of
carbohydrate metabolism are sensitive to nutritional stress in that they showed
decreased activity during starvation (De Bruin, 1976). Decrease in LDH enzymatic
activity level was observed in fishes when left in a frozen condition (Nambudiri and
Gopakumar, 1992). The activity of SDH in the tissues of control revealed the normal
aerobic capacity of the tissues. In the polluted group, the digestive gland , gill and
adductor muscle tissues showed decreased levels of SDH activity depicting the
hypoxic condition of these tissues. The haemolymph also showed the lower activity of
SDH contrast to that of control. The activity of SDH in these tissues of the polluted
also infers about the availability of the substrate namely the succinate and its further
oxidation in the Krebs cycle. Kabeer Ahammad et al. (1978) noticed a reduction in
the SDH activity in the foot, mantle and hepatopancreas of Lamellidens marginalis
Vitamin C or ascorbic acid is one among the several antioxidants such as vitaminE, Co-enzyme Q10, selenium, zinc etc., (Sinatra, 1998). using biocides. According to
Som et al. (1980) it protects the cellular membrane by inducing high stability.
Decrease in tissue ascorbic acid level due to pathogenesis was reported by Mitoma
and Smith (1960) and Gameel (1982). In the present study the ascorbic acid level was
found to be significantly lowered in all the tissues of the polluted zone forms over that
of the relatively unpolluted control group. Decreased levels of ascorbic acid affecting
germ cells in rainbow trout was reported by Ciereszko et al.(1999). In this context the
lower value of vitamin C estimated in the different tissues of P.viridis implicates upon
its consequences in its larval development, larval settlement, anchoring byssal thread
formation, adductor muscle development and finally the immunity in its bivalve.
Michael et al. (1998) reported that the fish Oreochromis mossambicus, when
administrated with multiple dose of ascorbic acid possessed insignificant
enhancement of antibody response. Martinez and Oliveira (2010) studied the bacterial
loading in Perna perna (Linnaeus, 1758) in the coastal waters of Brazil. A comparable
study in Malaysia showed that the mussels showed higher loads of bacterial coliform
(Sasikumar and Krishnamoorthy 2010). The above enzymatic and antioxidant changes
in this species by the pollution may lead to severe problem to these animals which can
be resolved by awareness to the human mankind.
10. CONCLUSION
Being sessile in habit the perna viridis is of importance, in the assessment of
environmental contamination. In the present study the survey of Perna viridis reveals that
the species is found in two zones viz., one in the shoreline area and the other in the
offshore area one kilometre away from the coast. The study has been proposed with a
view to delineating the difference between the two populations with reference to enzyme
composition, antioxidant content and microbial analysis. The bivalve mussel population
in natural marine habitats as well as in artificial laboratory situation shows that
environmental contaminations affect their normal development of adults, larval survival
and larval development. These studies revealing specific pathological changes in both
larvae and adults implicate upon their importance in environmental monitoring as
bioindicator organisms. The results of the present study and similar observations in the
adults of bivalves species elucidate the fact that environmental contamination could bring
deleterious effects which would cause morbidity and bring mortality of the population.
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