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05_ h. p. nandurkar.pmd
8(3): 997-1000, 2013 (Supplement on Toxicology)
Save Nature to Survive
STUDY OF TETRACYCLINE INDUCED ALTERATION IN ASCOR-
BIC ACID CONTENTS IN FRESHWATER BIVALVES, LAMELLIDENS
CORRIANUS (LEA) AND PARREYSIA CYLINDRICA (ANNANDALE
H. P. NANDURKAR
Department of Zoology, Sant Gadge Baba Amravati University, Amravati. (M.S.) 444 602, INDIA
e-mail: [email protected]
After exposure to lethal and sublethal concentrations of tetracycline, one of the broad spectrum antibiotics, the
vitamin C level showed the depletion in various tissues except in gill and foot of experimental models, the
freshwater bivalve, L. corrianus
and P. cylindrica.
The bivalves were exposed to acute dose of tetracycline (LC )
) 72.83 PPM up to 21 days. The ascorbic acid contents were
estimated in mantle, gill, foot, testis, ovary, whole body and digestive gland after 24 and 96h of acute and 7, 14
Received on :
and 21 days of chronic exposure. A lethal and sublethal concentration of tetracycline induced severe, progressive
perturbations in tissues after 96 hrs and 21 days. The level of ascorbic acid showed almost an overall decrease indifferent tissues but mainly in gill and mantle on lethal and sublethal exposures. In L. corrianus
the decrease in
Accepted on :
ascorbic acid was 22.22 % and 49.99% after 96 hrs and 21 days exposures respectively and in P. cylindrica
39.99% decrease after lethal dose induction of tetracycline. The foot and gill in case of P. Cylindrical
showedincreased ascorbic acid contents on lethal exposure and the rest tissues showed depleted level in the ascorbic acid
contents. In the foot of P. cylindrical
the level inclined up to 12.3% and 27.39 % when exposed to lethal and
sublethal concentration and in gill it was 16.12% after exposure of lethal dose.
this water soluble vitamin by their own, but not the humanbeing. Hence in case of human beings it is introduced through
The biochemical changes occurring in the body gives first
indication of stress. During any stress, to overcome the alteredsituation extra energy is needed. The biochemical composition
Ascorbic acid acts as an essential factor for normal growth in
varies according to seasonal changes, environmental factors
rainbow trout, Salmo gairdneri
(Halver et al
., 1969; Tucker,
(temperature, salinity), starvation and contamination of
1983; Tucker and Halver, 1986). In terrestrial animals the
pollutants due to different anthropogenic activities.
dietary ascorbic acid has role in the host defense systems.
Ascorbic acid influences various parameters of
The bivalves resist against such unwanted conditions by its
immunocompetence in the guinea pig (Thurman and
own way and try to minimize the effect of the altered situation
Goldstein, 1979). Though the complete prevention of viral
by removing the toxicant or made it simple by
infection is not possible, high doses of ascorbic acid reduces
biotransformation. Effects of toxicants result from their
potency of the viral diseases (Murata, 1975). The accumulation
interaction with certain receptors of the organisms. Thus the
of ascorbic acid at the site of wound healing was found by
impact of toxicant is exerted not only on cell but also on the
Gould (1963). Interferons get enhanced in circulatory system
cell content. Different toxicants affect the metabolic activities
after ascorbic acid ingestion through diet (Siegel, 1974).
which are expressed in terms of different changes that
Lymphoidal tissue regeneration and their differentiation
occurred in bivalve (Abel, 1974; Langston, 1986; Lumet et
occurred under the influence of ascorbic acid (Dieler and
., 2000). Ascorbic acid, being important constituent in cellular
Breitenbach, 1971). Siddique (1967) found the increase in
metabolism, the interactions of the bimolecular gives proper
ascorbic acid in liver, gonads and serum of Ophiocephalus
idea of toxicant stress and its effect.
with increase in temperature.
From the time ancient demand for the protein rich food, being
Thus ascorbic acid has a central position in curing the impaired
fulfilled by fishes, shellfishes, bivalves and oyster like sources,
condition occurred by the pathogenic attack and resists
as it provides many of the nutrient elements. The study of
against the diseases in organisms. The impact of tetracycline
these nutrients is possible by the biochemical analysis of the
on ascorbic acid content was studied in bivalves, L
different bimolecular in general and with the altering induced
and P. cylindrica
in the present study because in artificial
conditions in exposed animals in particular.
pearl culture during postoperative care bivalves are exposed
For different physiological acts vitamins are essential, although
to certain antibiotic treatment as there are more chances of
required in trace amount. Most of the animals can synthesize
bivalve mortality. Use of antibiotics reduces the rate of mortality.
Similarly in culture, to reduce the mortality rate of larvae of
The rest of the tissues depressed the contents at different
bivalves and oysters, antibiotics are used. So far, the side effects
proportions as compared to the control. The overall effect as
and toxic effects of antibiotics on the bivalves, oysters and
increase or decrease in ascorbic acid contents was significant
other invertebrates are not yet studied.
at P < 0.001, P< 0.01 or P < 0.05 level for acute and chronicexposure while was non-significant in some cases (Table 2 ).
MATERIALS AND METHODS
The antioxidant role of ascorbic acid is a well-knownphenomenon, which protects the tissues from the superoxide
The freshwater bivalves, L. corrianus
and P. cylindrica
radical generated due to different toxicological effects. Changes
collected from Girna dam, Dist: Nasik, M.S. The animals were
in the environment cause alteration in the ascorbic acid
acclimatized to laboratory conditions for 4 days prior to
experimentation. During experimentation only those animalsshowing movements and in apparent good health, were used
The varied functions of the ascorbic acid make it dynamic.
for investigation. The animals were divided into five groups,
Any alteration in the surrounding water due to the
two for acute and two for chronic exposures of tetracycline
contamination of water also alters ascorbic acid contents.
and one group was maintained as control in each case.
Different pollutant stress has its impact on the concentration
a) Acute exposure to Tetracycline
of ascorbic acid (Ali et al
., 1983; Bhusari, 1987). Ascorbic
The healthy bivalves, Lamellidens corrianus
were exposed to
acid contents increase during stress (Rao and Chinoy, 1986)
and after metal intoxication indicating its role in detoxification
were exposed to tetracycline 166.54PPM
The curing response against methyl mercury damage was seen
b) Chronic exposure to Tetracycline
in the reproductive organs of guinea pig after ascorbate
The acclimatized L. corrianus
were exposed to (LC
administration (Rao et al
., 1994). Seymour (1981, b) reported
concentration of tetracycline 73.82 PPM while P
that the levels of ascorbic acid in the ovaries of maturing crucian
were exposed to chronic concentration of tetracycline 33.30
carp, Carassius carassius
decreased after injection of pituitary
extract. Wedemeyer (1969) observed that the stress-induced
During exposure period, no special food was provided and
release of cortisol occurred concomitant with a decrease in
the water with required concentration of tetracycline was
the ascorbic acid in the kidney of salmonids.
changed daily in the experimental set. Control set was provided
In higher animals (vertebrates) the reduced exogenous
with dechlorinated water only without addition of tetracycline.
requirement of ascorbic acid may be a result of its lower need
After 24 and 96h of acute exposure and after every 7, 14 and
for biochemical functions with age or an increased storage
21 days of chronic exposure, the mantle, gill, foot, testis, ovary,
capacity combined with more efficient endogenous reuse.
digestive gland and the whole flesh were isolated, blotted to
Jadhav et al
. (1996) showed a decreased level of ascorbic
remove excess water and dried in oven at 80ºC till constant
acid content after pesticidal stress in Corbicula striatella
weight was obtained. All tissues were ground separately into
Waykar (2000) reported a decrease in the ascorbic acid level
fine powder from which ascorbic acid contents were estimated.
in various tissues of Parreysia cylindrica
on exposure to
Ascorbic acid content was estimated by using Hydrazine
reagent by the method as given by Roe (1967). The calibrationcurves were drawn by plotting concentrations of standard
Ascorbate protected the binding sites of the receptors and
against optical density to determine the corresponding value
making the metal forms insoluble (Scheuhanner and Cherian,
of ascorbic acid content from tissues after acute and chronic
1985). The rate of growth retardation caused by toxic metals
exposure to tetracycline. The results were expressed in mg
was minimized by the ascorbate administration (Hill, 1979).
per 100mg of dry tissue. The % variations were also calculated
Clarkson et al
. (1988) and Rao et al
. (1994) reported that the
to find out the antibiotic induced stress to the biochemical
oxygen radical formed due to methyl mercury forms reactive
substances undertaken for study and the test of significance
oxygen intermediates with ascorbic acid. Daine et al
showed the recovery from chromium intoxication by ascorbicacid treatment. Sometimes vitamin C and vitamin E acts in
RESULTS AND DISCUSSION
combination for detoxification (Chan, 1993; Meister, 1994).
Mahajan and Zambare (2001) found that the reduction in
The maximum ascorbic acid content was observed in mantle.
protein depletion due to CuSO and HgCl was recovered by
There was a marked decrease in ascorbic acid contents in
ascorbate treatment in Corbicula striatella
almost all tissues of L. corrianus
and P. cylindrica
Mouse peritoneal macrophages when elicited by the
and chronic exposures. The significant effect of tetracycline
antioxidant ascorbic acid have been found to be significantly
observed in gills of L. corrianus
was 49.99 % and the reverse
stimulatory, exhibiting significant enhancement in protein
trend was shown by the whole body 33.33 % increase in
content, lysosomal acid hydrolase levels and capability to
ascorbic acid contents after sublethal exposure of tetracycline
phagocytise (Agrawal et al
., 2003). The ascorbic acid supply
may boost the macrophage activity, helping to remove
After 96h of lethal exposure to tetracycline showed incline
intracellular free irritant. These results indicate the positive
pattern of ascorbic acid content in gill and foot 16.12% and
role of ascorbic acid in toxicant stress.
12.30 % respectively while sublethal exposure increased the
The depleted level of ascorbic acid is a vivid response against
contents in foot up to 27.39 % after 21 days in P. cylindrica
tetracycline to cope up the toxic stress caused by exposure to
Table 1: Impact of Tetracycline on ascorbic acid content of Lamellidens corrianus after acute and chronic exposure
Table 1: Cont.
M = Mantle; G = Gill; F = Foot; O = Ovary; T = Testis; WB = Whole body; DG = Digestive gland; Values are expressed as mg/100mg dry weight of tissue. ± indicates standard deviationof three independent replications; + or - indicates % variation over control. Significance: * p< 0.05; ** p < 0.01; *** p 0.001; NS = Non-significant
Table 2: Impact of Tetracycline on ascorbic acid content of Parreysia cylindrica after acute and chronic exposure
Table 2: Cont.
0.675±0.05817+15.151NS 0.3733±0.0290 0.4382±0.0101+17.391*** 0.4088±0.02936
0.2133±0.0050-33.333*** 0.3911±0.0029 0.2666±0.00-31.818***
M = Mantle; G = Gill; F = Foot; O = Ovary; T = Testis; WB = Whole body; DG = Digestive gland; Values are expressed as mg/100mg dry weight of tissue. ± indicates standard deviationof three independent replications; + or - indicates % variation over control. Significance: * p < 0.05; ** p < 0.01; *** p 0.001; NS=Non-significant
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