2.7.5 Reduced Glutathione (GSH)
Reduced glutathione levels were determined by following the method of Ganie et al.
[62]. In brief, the tissues were homogenized in 100 mL of ice-cold potassium
phosphate buffer (0.1 M, pH 6.5) followed by adding 100 mL of 25% TCA to
precipitate the homogenate. Then the precipitate was centrifuged at 3,000 g for
10 min, 4
C, and 150 mL of supernatant were transferred to aliquots containing
reaction mixture (60 mM DTNB and 450 mL of 50 mM potassium phosphate buffer
(pH 7.4)). The absorbance of the samples was read at 412 nm in UV
spectrophotometer.
2.8 Statistical Analysis
A statistical analysis was performed using SPSS software – Ver.16 statistical
package. The results of ASA -treated groups were compared against the control
followed by one-way ANOVA and the Duncan’s multiple range test at p < 0.05
level.
3 Results
3.1 Haematology
The changes in the Hb level of fish C. carpio exposed to chronic concentrations
(100 mg L
À1 - Treatment I and 200 mg L
À1 - Treatment II) of acetylsalicylic acid
(ASA) for a period of 21 days were illustrated in Fig. 1. During the above exposure
period, Hb level was decreased in both the treatments throughout the study period. A
similar trend was also noted for Ht and RBC count in ASA-treated group when
compared with their respective control groups (P < 0.05). A maximum percent
decrease of Hb level was recorded at the end of 14th day in Treatment II (Fig. 2).
Similarly a maximum percent decrease of HCT level was noted on 14th day in
Treatment II. There was a significant (P < 0.05) decrease in the RBC count at the
end of 7th, 14th and 21st days of exposure in ASA-treated groups when compared to
control groups (Fig. 3). In both the treatments, WBC level of C. carpio was elevated
when compared to the control group (Fig. 4). Compared with the control groups,
ASA-treated Cyprinus carpio had significantly higher MCV (Fig. 5), MCH (Fig. 6)
and MCHC values (Fig. 7) throughout the study period.
Toxicity Assessment of Acetylsalicylic Acid to a Freshwater Fish Cyprinus. . .
199
Reduced glutathione levels were determined by following the method of Ganie et al.
[62]. In brief, the tissues were homogenized in 100 mL of ice-cold potassium
phosphate buffer (0.1 M, pH 6.5) followed by adding 100 mL of 25% TCA to
precipitate the homogenate. Then the precipitate was centrifuged at 3,000 g for
10 min, 4
C, and 150 mL of supernatant were transferred to aliquots containing
reaction mixture (60 mM DTNB and 450 mL of 50 mM potassium phosphate buffer
(pH 7.4)). The absorbance of the samples was read at 412 nm in UV
spectrophotometer.
2.8 Statistical Analysis
A statistical analysis was performed using SPSS software – Ver.16 statistical
package. The results of ASA -treated groups were compared against the control
followed by one-way ANOVA and the Duncan’s multiple range test at p < 0.05
level.
3 Results
3.1 Haematology
The changes in the Hb level of fish C. carpio exposed to chronic concentrations
(100 mg L
À1 - Treatment I and 200 mg L
À1 - Treatment II) of acetylsalicylic acid
(ASA) for a period of 21 days were illustrated in Fig. 1. During the above exposure
period, Hb level was decreased in both the treatments throughout the study period. A
similar trend was also noted for Ht and RBC count in ASA-treated group when
compared with their respective control groups (P < 0.05). A maximum percent
decrease of Hb level was recorded at the end of 14th day in Treatment II (Fig. 2).
Similarly a maximum percent decrease of HCT level was noted on 14th day in
Treatment II. There was a significant (P < 0.05) decrease in the RBC count at the
end of 7th, 14th and 21st days of exposure in ASA-treated groups when compared to
control groups (Fig. 3). In both the treatments, WBC level of C. carpio was elevated
when compared to the control group (Fig. 4). Compared with the control groups,
ASA-treated Cyprinus carpio had significantly higher MCV (Fig. 5), MCH (Fig. 6)
and MCHC values (Fig. 7) throughout the study period.
Toxicity Assessment of Acetylsalicylic Acid to a Freshwater Fish Cyprinus. . .
199