The interactive effect of elevated temperature on deltamethrin-induced biochemical stress responses in Channa punctata Bloch

The interactive effect of elevated temperature on deltamethrin-induced biochemical stress responses in Channa punctata Bloch

Chemico-Biological Interactions 193 (2011) 216–224 Contents lists available at SciVerse ScienceDirect Chemico-Biological Interactions journal homepa...

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Chemico-Biological Interactions 193 (2011) 216–224

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Chemico-Biological Interactions journal homepage: www.elsevier.com/locate/chembioint

The interactive effect of elevated temperature on deltamethrin-induced biochemical stress responses in Channa punctata Bloch Manpreet Kaur a, Fahim Atif b, Rizwan A. Ansari a, Firoz Ahmad a, Sheikh Raisuddin a,⇑ a b

Department of Medical Elementology and Toxicology, Hamdard University (Jamia Hamdard), New Delhi 110062, India Brain Research Laboratory, Department of Emergency Medicine, Emory University, Atlanta, GA, USA

a r t i c l e

i n f o

Article history: Received 11 February 2011 Received in revised form 3 June 2011 Accepted 30 June 2011 Available online 23 July 2011 Keywords: Deltamethrin Heat stress HSP Antioxidant enzymes Oxidative stress

a b s t r a c t There are reports showing interactive effect of environmental factors with the toxic outcome of chemicals. We studied the interactive effect of elevated temperature as an abiotic stressor on deltamethrininduced biochemical stress responses in a freshwater fish, Channa punctata Bloch. Heat stress (12 °C above ambient temperature for 3 h) and pesticide exposure (deltamethrin 0.75 ppb for 48 h) showed significant induction of heat shock protein-70 (HSP70) in liver, kidney and gills of fishes. Elevated temperature when followed by deltamethrin exposure showed synergistic effect showing a high level of HSP70 in liver and gills whereas response in the kidney was opposite. On the contrary, when deltamethrin exposure followed the heat stress, no significant difference was observed. Protein carbonylation was found to be more pronounced in heat-stressed group compared with control fish group. A significant increase in lipid peroxidation (LPO) was observed in different tissues of fish exposed to either of the stressors. In the kidney of fish exposed to heat stress followed by deltamethrin, LPO was relatively lower as compared to other treatments. Thiols content such as reduced glutathione (GSH), total thiols (T-SH), nonprotein thiols (NP-SH) and protein thiols (P-SH) showed no consistent pattern in different tissues. In deltamethrin-exposed group that was subsequently exposed to heat stress, the GSH content was higher in liver and lower in both kidney and gills when compared with other groups. Alteration in the activities of antioxidant enzymes such as catalase (CAT), glutathione S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx) was also observed when fish were exposed to heat stress and/or deltamethrin. Our study demonstrated that heat stress modulated biochemical stress responses in fish showing a tissue specific pattern. This implies that fish has the capacity to elicit differential response to exposure to abiotic stressors in order to reduce the systemic magnitude of stress which may otherwise lead to severe dysfunction of vital tissues. Ó 2011 Elsevier Ireland Ltd. All rights reserved.

1. Introduction Temperature is an important environmental variable that has reflective impact on aquatic ectotherms, such as fishes. Stress responses including those induced by temperature variation, dictate whether the organism adapts, survives, or dies [1]. Higher water temperature lowers the availability of dissolved oxygen (DO), accelerates the metabolism, alters the respiration rate and enhances the oxygen demand of fish. Fishes are poikilotherms Abbreviations: CAT, catalase; BOD, biological oxygen demand; COD, chemical oxygen demand; DO, dissolved oxygen; GSH, reduced glutathione; GST, glutathione S-transferase; GR, glutathione reductase; GPx, glutathione peroxidase; HSP, heat shock protein; LPO, lipid peroxidation; T-SH, total thiols; TSS, total suspended solids; NP-SH, non-protein thiols; P-SH, protein thiols. ⇑ Corresponding author. Tel.: +91 11 26059688; fax: +91 11 26059663. E-mail addresses: [email protected], [email protected] (S. Raisuddin). 0009-2797/$ - see front matter Ó 2011 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.cbi.2011.06.011

and change in water temperature has a great impact on their physiology [2–5]. Gradual change in temperature can be physiologically compensated for, but a rapid change disturbs homeostasis and thus becomes a stress. Pesticides are chemical stressors, which have become an integral part of the ecosystem and regarded as major environmental pollutants over the years. When used in the vicinity of aquatic ecosystems, they tend to mix with water and may exert adverse effects on fish populations [6,7]. Pyrethroids including deltamethrin are being used as substitutes for organochlorines and organophosphates. They are extensively used in agriculture, for controlling pests, insects and vectors of endemic diseases, protecting seeds during storage and fighting household insects because of their low environmental persistence [8]. The potential hazard of pyrethroids is owed to their heavy use in many aquatic larvicidal and malarial control programs. Deltamethrin and other pyrethroids exposure have been found to be highly toxic to fish and its

M. Kaur et al. / Chemico-Biological Interactions 193 (2011) 216–224

oxidative stress-inducing effect has been demonstrated in fish under short-term exposure [9,10], thus warranting a thorough ecotoxicological risk assessment [11–14]. Fishes can exhibit a cumulative response to stressors [15]. When stressors, singularly or in combination, are severe enough to challenge the homeostatic mechanisms beyond the compensatory limits of fish or permanently alter them, physiological processes generally adapt to compensate for the stress [16]. Heat shock proteins (HSPs) are a family of proteins that are expressed in response to a diverse range of biotic and abiotic stressors. They are thus, also referred to as stress proteins. Stress protein induction has been attributed to many environmental pollutants in aquatic organisms. However, there is a paucity of data concerning HSPs and pesticide stress, with the exception of a few recent studies of paraquat [17], heptachlor exposure in Homarus americanus larvae [18] and rainbow trout exposed to deltamethrin [19]. The physiology of fish is strongly related to temperature which is supported by studies which have shown their response to chemical exposures is influenced by ambient temperature [1,5,15]. The interactive effect may be additive, synergistic or antagonistic and the nature of the interaction may differ for different compounds or stressors. Temperature has also been suggested to play an important role in the toxicity of aquatic metals [20]. Additionally, exposures to pesticides in combination with other agents (viz., stressors) may exert effects different from those experienced with pesticides alone [21]. It is well established that stressors can elicit non-specific responses in fish, which are considered adaptive and hence enable the fish to cope with the disturbance and maintain its homeostatic state. Antioxidant defense systems with non-enzymatic and enzymatic components are altered by stresses and are well documented [22]. Aquatic organisms including fish are routinely exposed to a mixture of aquatic pollutants; we were interested in studying the effect of temperature and deltamethrin on stress responses in fish. In the last decade, deltamethrin use has exponentially increased in some countries, like, India [23], where the seasons are distinct, leading to substantial variation in temperature. Such environmental variation of temperature may considerably influence the toxicity of pesticides. While the effects of temperature and contaminants on fish have been studied as individual stressors, there is no detailed information about the combined effects of such stressors. With this background the present research was undertaken to study stress responses of Channa punctata Bloch on interaction of elevated temperature and deltamethrin exposure.

2. Materials and methods 2.1. Fish C. punctata Bloch (Spotted snake-head murrel, Order: Perciformes, Family: Channidae) weighing 50–75 g were commercially procured and maintained in 60 l aquaria following standard procedure [24]. Aquarium water was kept oxygen saturated by continuous aeration and ambient aquarium temperature was maintained at 20 ± 2 °C with a photoperiod of 12 h light and 12 h dark cycle. Fish were acclimatized for 2 weeks under the above mentioned conditions before the start of the experimental paradigm. Physico-chemical characteristics of aquarium water were monitored every alternate day. The normal ranges of selected parameters were as follows: DO = 7.1 ± 0.8 mg/l, biological oxygen demand (BOD) = 9.7 ± 0.2 mg/l, chemical oxygen demand (COD) = 14.9 ± 1.1 mg/l, total suspended solid (TSS) = 8.0 ± 0.9 mg/l, turbidity (in NTU) = 5 ± 0.2 and pH 7.6 ± 0.08. Aquarium water was replaced every 24 h to minimize contamination from metabolic wastes. Fish

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were handled using the institutional animal ethical committee (IAEC) guidelines of animal usage. 2.2. Exposure groups Acclimatized fish were divided into five groups each comprised a minimum of eight fish. The first group maintained at ambient temperature 20 ± 2 °C served as control (C). The second group of fish was exposed to 0.75 ppb deltamethrin (in acetone) in aquarium water for 48 h (DEL). Fish of the third group were exposed to pre-heated water (32 °C; 12 °C above the average ambient temperature) for 3 h (HS). The fourth group comprised of the fish which were first exposed to elevated temperature for 3 h and then to deltamethrin (0.75 ppb for 48 h) (HS + DEL). Whereas, the fish of the fifth group were first exposed to deltamethrin and then to heat stress at the same concentration and temperature as mentioned above (DEL + HS). Experiment was designed in such a manner that all the fish were sacrificed at the same time. The dose concentration of deltamethrin was selected on the basis of previously published reports on C. punctata [9,10]. 2.3. HSP measurement Stress protein HSP70 was analyzed in different tissues of C. punctata using indirect ELISA procedure [25]. SDS–PAGE was also performed to study induction of HSP70 in different tissues [26]. 2.3.1. ELISA Each well of 96-well microtiter plate (Nunc, USA) was coated with sample (50 lg protein) in triplicate and incubated at 4 °C overnight. After incubation, each well was washed thrice with Tween-20 in PBS (TPBS, pH 7.2). After washes, plate was blocked with 2% bovine serum albumin (BSA, Sigma) for 1 h at 37 °C. After blocking, wells were again washed and 100 ll anti-HSP70 primary antibody (1:4000, anti-HSP70 [5A5, ab2787] mouse monoclonal from Abcam Ltd., UK) was added to each well and incubated at 37 °C. Peroxidase tagged anti mouse IgG (1:4000 dilution) was used as secondary antibody (Bangalore Genei Pvt. Ltd.) and the antigen–antibody reaction was developed by adding substrate solution (0.1% tetramethylbenzidine in phosphate citrate buffer, pH 5.0) to each well. The plate was incubated at 37 °C for 2 min and yellow color developed was read at 450 nm in Microplate Reader (Benchmark, Biorad) after stopping the reaction with 50 ll of 2 M H2SO4. 2.3.2. SDS–PAGE Tissue samples were homogenized after thawing in 66 mM Tris buffer (pH 7.8, Hi-Media Labs, Mumbai, India) with 1% Igepal CA 630, 0.1 mM phenylmethylsulfonyl fluoride (PMSF) and protease inhibitor cocktail (Sigma) to inhibit proteases. Samples were transferred to microcentrifuge tubes and SDS sample buffer (63 mM Tris–HCl, pH 6.8, 10% glycerol, 2% SDS, 0.0025% bromophenol blue; Hi-Media Labs) was added to each sample tube and heated at 95– 100 °C for 5 min. The proteins were separated by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE, Biorad Mini Protein II) using 12.5% sodium dodecyl sulfate–polyacrylamide gel with a 5% stacking gel as described by Laemmli [27], at 100 V for 1 h. Samples in duplicate were then run simultaneously on gels. One was stained using Coomassie brilliant blue stain (E-Merck, Germany) and other was used for blotting. 2.4. Preparation of homogenate and post-mitochondrial supernatant (PMS) After end of the treatment schedule fish were sacrificed and dissected to remove liver, kidney and gills. Tissues were washed in

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ice-cold saline (0.85% NaCl) and for gills, gill rakers were removed and lamellae were homogenized. A 10% (w/v) tissue homogenate was prepared in chilled phosphate buffer (0.1 M, pH 7.4) containing KCl (1.17%) using a Potter–Elvehjem homogenizer. The homogenate was centrifuged in a refrigerated centrifuge (Hermle, Model Z323 K) at 800g for 5 min at 4 °C to separate the nuclear debris. The supernatants were again centrifuged at 10,500g for 30 min at 4 °C to obtain the PMS for various biochemical analyses. 2.5. Protein carbonyl assay Protein carbonyl content was assayed by the procedure of Floor and Wetzel, [28]. Soluble protein (0.5 ml) reacted with 10 mM 2,4dinitrophenylhydrazine (DNPH) in 2 M hydrochloric acid for 1 h at room temperature and precipitated with 6% trichloroacetic acid (TCA). The pelleted protein was washed thrice by resuspension in ethanol/ethyl acetate (1:1). Proteins were then solubilized in 6 M guanidine hydrochloride, 50% formic acid and centrifuged at 16,000 g for 5 min to remove any trace of insoluble material. The carbonyl content was measured spectrophotometrically (Shimadzu UV–Vis spectrophotometer, Japan) at 366 nm. Assay was performed in triplicate and a tissue blank incubated with 2 M HCl without DNPH was included for each sample. The results were expressed as nmol of carbonyl/mg protein based on the molar extinction coefficient of 21,000 M1 cm1[28]. 2.6. LPO LPO was measured by the procedure of Mihara and Uchiyama, [29] with some modifications. Briefly, 0.25 ml of homogenate was mixed with 25 ll of 10 mM butylated hydroxytoluene (BHT, Sigma), 3 ml of 1% o-phosphoric acid (OPA, CDH Chemicals, Mumbai, India) and 1 ml of 0.67% thiobarbituric acid (TBA, Hi-Media Labs) were added, and mixture was incubated at 90 °C for 45 min. The absorbance was measured at 535 nm. The rate of LPO was expressed as nmol of TBA reactive substance (TBARS) formed/h/g of tissue using a molar extinction coefficient of 1.56  105 M1 cm1.

2.8. Antioxidant enzymes Catalase (CAT) activity was measured by the method of Claiborne, [33]. The assay mixture consisted of 1.95 ml phosphate buffer (0.05 M, pH 7.0), 1 ml hydrogen peroxide (0.019 M H2O2, CDH chemicals) and 0.05 ml PMS in a final volume of 3 ml. CAT activity was measured at 240 nm and calculated as nmol H2O2 consumed/ min/mg protein. Glutathione S-transferase (GST) activity was measured by the method of Habig et al. [34]. The reaction mixture consisted of 1.65 ml phosphate buffer (0.1 M pH 6.5), 0.1 ml GSH (1 mM, Sigma), 0.05 ml 1-chloro-2,4-dinitrobenzene (1 mM CDNB, Sigma) and 0.2 ml PMS in a total volume of 2 ml. The enzyme activity was measured at 340 nm and calculated as nmol CDNB conjugates/min/mg protein using a molar extinction coefficient of 9.6  103 M1 cm1. Glutathione peroxidase (GPx) activity was assayed according to the method described by Mohandas et al. [35] with some modifications. The assay mixture consisted of 1.44 ml phosphate buffer (0.1 M, pH 7.6), 0.1 ml ethylene diamine tetra-acetic acid (1 mM, EDTA, Sigma), 0.1 ml sodium azide, 0.05 ml glutathione reductase 1 IU/ml (GR, Sigma), 0.1 ml GSH (1 mM), 0.1 ml nicotinamide adenine dinucleotide phosphate reduced (0.2 mM, NADPH, Sigma), 0.01 ml H2O2 (0.25 mM) and 0.1 ml PMS in a total volume of 2 ml. The enzyme activity was measured at 340 nm and calculated as nmol NADPH oxidized/min/mg of protein, using a molar extinction coefficient of 6.22  103 M1 cm1. The glutathione reductase (GR) activity was measured by the method of Pandey et al. [36]. The reaction mixture consisted of 1.6 ml phosphate buffer (0.1 M, pH 7.4), 0.1 ml EDTA (0.5 mM), 0.1 ml GSSG (1 mM, Sigma), 0.1 ml NADPH (0.1 mM) and 0.1 ml PMS in a total volume of 2 ml. The enzyme activity was quantitated at 25 °C by measuring the disappearance of NADPH at 340 nm and calculated as nmol NADPH oxidized/min/mg protein using a molar extinction coefficient of 6.22  103 M1 cm1. All the enzymatic assays were monitored for 3 min and the difference of OD per min was used to calculate the respective activities against non-enzymatic blank consisting of all the reagents except PMS and making up the volume using phosphate buffers.

2.7. Estimation of reduced glutathione and thiols Reduced glutathione was determined in PMS by the method of Jollow et al. [30]. Sulfosalicylic acid 4% in the ratio of 1:1 was used to precipitate PMS. The samples were kept at 4 °C for 1 h followed by centrifugation at 400g for 15 min at 4 °C. The assay mixture consisted of PMS, phosphate buffer (0.1 M, pH 7.4) and dithiobis-2-nitrobenzoic acid, (DTNB, Sigma) in the total volume of 3 ml. The optical density of reaction product was read immediately at 412 nm on a spectrophotometer. The GSH values were calculated as lmol GSH/g tissue. Total thiol (T-SH), protein bound thiol (P-SH) and non-protein bound thiol (NP-SH) groups in the PMS were determined using the method of Sedlak and Lindsay [31], as adopted by Parvez et al. [32] in case of fish. For total thiols, 1.5 ml Tris buffer (0.2 M, pH 8.2), 0.1 ml 0.01 M DTNB, PMS and methanol were mixed in the total volume of 10 ml. After 10 min, mixture was centrifuged at 3000g at 4 °C for 10 min and the absorbance of the supernatant was measured at 412 nm. For non-protein thiols, 0.4 ml PMS was precipitated with 0.1 ml 40% TCA and 0.5 ml distilled water. After 10 min, mixture was centrifuged at 3000g. The absorbance of supernatant with tris buffer (0.4 M, pH 8.9) and 0.01 M DTNB in the total volume of 1.5 ml was read at 412 nm. The level of P-SH was calculated by subtracting the values of NPSH from T-SH content. The molar extinction coefficient of 13,000 M1 cm1 was used to measure various thiols. The values are expressed as lmol/g of wet tissue.

2.9. Protein estimation Protein content in various samples was estimated by the method of Lowry et al. [37] using Folin reagent (Sigma) and BSA as standard. 2.10. Statistical Analysis Analysis of variance (ANOVA) was applied to determine significant differences between data of different groups when compared with control. p values <0.05 were considered significant. Subsequently, Students–Newman–Keul’s test was applied for analyzing the significant difference between different treatment groups. The values are expressed as means ± SE. 3. Results 3.1. Effect on HSP70 Deltamethrin at the dose of 0.75 ppb for 48 h and heat stress for 3 h both induced HSP70 in all the tissues of C. punctata as measured by ELISA (Fig. 1). A noticeable induction of amount of constitutive HSP70 was also noted in all the tissues analyzed. HSP levels in liver increased significantly (p < 0.01) with more or less in same manner in all the groups as compared to control but there were no significant differences recorded among combined exposures when

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A

A

B

C

D

B

C

D

E

F

116.0 66.2

B

A

E

F

116.0 66.2

C

A

B

C

D

E

F

116.0 66.2

Fig. 2. SDS–PAGE image showing protein profile of fish liver (A), kidney (B) and gills (C) after exposure to heat stress (3 h) and deltamethrin 0.75 ppb for 48 h only and in combined exposure followed one by the other. Lane A was loaded with molecular weight protein marker. Lane B is control, Lane C exposed to deltamethrin alone and Lane D to heat stress only. Lanes E and F of combined exposure of deltamethrin followed by heat stress and vice versa, respectively. Arrow indicates HSP70 (stress proteins).

Table 1 Protein carbonyl content in different tissues of Channa punctata Bloch following exposure to deltamethrin and heat stress. Groups

Fig. 1. HSP70 in fish Channa punctata liver (A), kidney (B) and gills (C) after exposure to heat stress (3 h) and deltamethrin 0.75 ppb for 48 h only and in combined exposure followed one by the other. The values are expressed as in terms of relative optical density at 490 nm in ELISA (means ± SE, n = 5). Significant difference is shown as bp < 0.01 when compared with controls; pp < 0.05 and q p < 0.01 when compared with heat stressed (3 h) group and yp < 0.01 when compared with deltamethrin (0.75 ppb x 48 h) only exposed group.

compared to the deltamethrin exposed and heat stressed group. In kidney, although the HSP levels were significantly (p < 0.05–0.01) higher in the groups with stress of deltamethrin or heat, the values of HSP70 in the combined stressed groups were relatively lower than those with either of the stressors. Induction of HSP in kidney was found to be significantly (p < 0.01) lower in the group where heat stress followed deltamethrin exposure when compared to deltamethrin exposed and heat stressed groups. In gills of fish, heat stress induced significantly (p < 0.01) more HSP than deltamethrin and values were still significantly higher (p < 0.05–0.01) for groups that received combined exposure when compared to the groups that received exposure, either of heat stress or deltamethrin alone. The results were also supported by the SDS–PAGE profile of different tissue samples (Fig. 2).

3.2. Effect on protein carbonyl groups A significant (p < 0.05–0.01) rise in the content of protein carbonyls in liver, kidney and gills was observed in groups exposed to deltamethrin, heat exposure and combination of both the stressors as compared to that of controls (Table 1). Protein carbonylation

Control Heat Stress (HS) Deltamethrin (DEL) HS + DEL DEL + HS

Tissues Liver

Kidney

Gills

0.9 ± 0.06 2.3 ± 0.16b 1.4 ± 0.02b 1.7 ± 0.02b,q,x 2.0 ± 0.03b, x

1.4 ± 0.09 2.2 ± 0.12b 2.4 ± 0.13b 2.4 ± .09b 1.9 ± 0.06b,x

1.6 ± 0.09 2.9 ± 0.14b 2.2 ± 0.07b 2.0 ± 0.04a,q 2.4 ± 0.10b

Values are expressed as nmol of carbonyl/mg protein (means ± SE, n = 5). Significant difference is shown as ap < 0.05, bp < 0.01 when compared with controls; qp < 0.01 when compared with heat stressed (3 h) group and xp < 0.05, when compared with deltamethrin (0.75 ppb  48 h) only exposed group.

was more prominent in heat stressed group as compared to deltamethrin-exposed group. In the group of fish where heat stress followed the exposure by deltamethrin or vice versa showed decreased carbonylation of proteins as compared to only heat stressed group. 3.3. Effect on LPO Sequential exposures to deltamethrin for 48 h at the dose of 0.75 ppb and heat stress for 3 h induced a significant (p < 0.05– 0.01) increase in LPO in all the tissues when compared with control fish (Fig. 3). Similar responses were observed with the combined exposure of the two stresses in the study, one following the other. In liver, it increased LPO levels significantly (p < 0.01) in all cases as compared to controls. In kidney also, the response was same except in the group of fish where heat stress followed deltamethrin exposure, the significant increase (p < 0.05) in LPO was relatively lower as compared to other treatments. LPO response was noteworthy in gills where significant increase (p < 0.05) in the production of TBARS was observed for group exposed to deltamethrin only and the group with deltamethrin exposure followed by heat stress when compared to fish with heat stress only.

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M. Kaur et al. / Chemico-Biological Interactions 193 (2011) 216–224 Table 2 Non-enzymatic antioxidants as modulated by heat stress and deltamethrin in the liver of Channa punctata Bloch. Groups

Control Heat Stress (HS) Deltamethrin (DEL) HS + DEL DEL + HS

Parameters Total thiols

Non-protein thiols

Protein thiols

72.5 ± 3.8 86.8 ± 2.0a 90.1 ± 5.4a 119.8 ± 6.5b,q,x 130.8 ± 4.2b,q,y

1.3 ± 0.04 1.1 ± 0.04a 1.7 ± 0.1b 1.2 ± 0.04a,y 1.4 ± 0.04q,y

71.2 ± 3.8 85.6 ± 1.9a 88.3 ± 5.3a 118.7 ± 6.5b,q,x 129.4 ± 4.2b,q,y

Values are expressed as lmol/g of wet tissue (means ± SE, n = 5). Significant difference is shown as ap < 0.05, bp < 0.01 when compared with controls; qp < 0.01 when compared with heat stressed (3 h) group and xp < 0.05, yp < 0.01 when compared with deltamethrin (0.75 ppb  48 h) only exposed group.

Table 3 Non-enzymatic antioxidants as modulated by heat stress and deltamethrin in the kidney of Channa punctata Bloch. Fig. 3. LPO level in different tissues of Channa punctata after exposure to heat stress (3 h) and deltamethrin (0.75 ppb for 48 h) individually (HS and DEL, respectively) or in combination when heat treatment was given either before or after deltamethrin exposure (HS + DEL and DEL + HS, respectively). Values are expressed as nmol of TBARS formed/h/g of tissue (means ± SE, n = 5). Significant difference in result is shown on bar as ap < 0.05 and bp < 0.01 when exposure group data were compared with respective controls.

3.4. Effect on thiol profile 3.4.1. Effect on GSH Both the stresses under study (deltamethrin and heat stress) in combination or alone significantly altered the GSH content in various tissues (Fig. 4). Heat stress for 3 h significantly (p < 0.01) decreased the GSH content in all the tissues, whereas deltamethrin exposure significantly (p < 0.05–0.01) increased the content of GSH in liver, kidney and gills when compared to controls. Fish exposed to deltamethrin after heat stress showed significant (p < 0.01) increase and decrease in quantity of GSH in kidney and gills, respectively whereas no significant changes were observed

Groups

Control Heat Stress (HS) Deltamethrin (DEL) HS + DEL DEL + HS

Parameters Total thiols

Non-protein thiols

Protein thiols

89.3 ± 1.9 76.8 ± 1.3b 118.3 ± 8.7a 106.6 ± 7.0a,q 81.7 ± 1.8a,q,y

1.6 ± 0.2 1.2 ± 0.1a 2.2 ± 0.1a 1.7 ± 0.1p,x 1.1 ± 0.1a,y

87.7 ± 1.8 75.7 ± 1.3 116.1 ± 8.6a 104.9 ± 6.9a,q 80.6 ± 1.7a,q,y

Values are expressed as lmol/g of wet tissue (means ± SE, n = 5). Significant difference is shown as ap < 0.05, bp < 0.01 when compared with controls; pp < 0.05, q p < 0.01 when compared with heat stressed (3 h) group and xp < 0.05, yp < 0.01 when compared with deltamethrin (0.75 ppb  48 h) only exposed group.

Table 4 Non-enzymatic antioxidants as modulated by heat stress and deltamethrin in the gills of Channa punctata Bloch. Groups

Control Heat Stress (HS) Deltamethrin (DEL) HS + DEL DEL + HS

Parameters Total thiols

Non-protein thiols

Protein thiols

64.1 ± 3.6 79.1 ± 1.8a 50.6 ± 0.9a 65.4 ± 2.2q,y 69.2 ± 3.6p,y

0.6 ± 0.04 0.4 ± 0.03b 0.8 ± 0.05a 0.7 ± 0.09a,q,x 0.7 ± 0.01b,q

63.5 ± 3.6 78.7 ± 1.7a 49.8 ± 0.9a 64.7 ± 2.3q,y 68.5 ± 3.6p,y

Values are expressed as lmol/g of wet tissue (means ± SE, n = 5). Significant difference is shown as ap < 0.05, bp < 0.01 when compared with controls; pp < 0.05, q p < 0.01 when compared with heat stressed (3 h) group and xp < 0.05, yp < 0.01 when compared with deltamethrin (0.75 ppb  48 h) only exposed group.

in liver. In deltamethrin-exposed group that was subsequently stressed by heat, the GSH content was significantly (p < 0.01) higher in liver and significantly (p < 0.05–0.01) lower in both kidney and gills when compared to deltamethrin and heat stressed groups alone along with the control group.

Fig. 4. GSH content in different tissues of fish Channa punctata after exposure to heat stress (3 h) and deltamethrin 0.75 ppb for 48 h only and in combined exposure followed one by the other. The values are expressed as lmol of GSH per g of tissue (means ± SE, n = 5). Significant difference is shown as ap < 0.05, bp < 0.01 when compared with controls; pp < 0.05, qp < 0.01 when compared with heat stressed group and xp < 0.05, yp < 0.01 when compared with deltamethrin only exposed group.

3.4.2. Effect on T-SH, NP-SH and P-SH Levels of T-SH, NP-SH and P-SH were altered significantly (p < 0.05–0.01) in liver, kidney and gills of all the stressed fish groups. T-SH content increased significantly (p < 0.05–0.01) in all the tissues with exception in kidney where it decreased significantly (p < 0.05–0.01) with heat stress and in gills of fish with deltamethrin exposure. Dual exposure of stressors (heat and deltamethrin) in liver significantly (p < 0.05–0.01) increased T-SH content when compared with controls and single stresses of deltamethrin and heat whereas the contents were unchanged in gills. NP-SH almost shared the pattern similar to GSH in all the tissues,

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where it decreased significantly (p < 0.05–0.01) with heat stress and increased significantly (p < 0.05–0.01) with deltamethrin exposure. Groups in which one stress followed the other, showed significant changes when compared to groups with their respective stressor in alone. Pattern of P-SH modulation was similar to that found with T-SH (Tables 2–4). 3.5. Effect on antioxidant enzymes Antioxidant enzyme activities (Tables 5–7) displayed differential but significant (p < 0.05–0.01) responses to single and combined exposures of deltamethrin and heat stress. With the exception of significant (p < 0.01) increase in liver in response to heat stress, CAT activity was significantly (p < 0.05–0.01) decreased in response to heat stress and deltamethrin in rest of the tissues as compared to controls. The values in liver were still significantly (p < 0.05–0.01) lower in the groups where one stress was followed by another when compared to groups that received single stress. In kidney, groups with combined stress showed no changes as com-

pared to controls but the values were significantly (p < 0.05–0.01) higher to their respective stresses. In gills the group that was exposed to deltamethrin after heat stress showed significantly (p < 0.05–0.01) higher CAT activity than the group that received deltamethrin and heat stress exposure only. GST activities with heat stressed fish were significantly (p < 0.05) decreased whereas in deltamethrin exposure the activities were significantly (p < 0.05–0.01) higher in liver and kidney but lower (p < 0.01) in the gills in all the exposures (Tables 5–7). When heat stress followed deltamethrin stress the activity of GST in liver and kidney were significantly (p < 0.05–0.01) increased in contrast to gills where it significantly (p < 0.05–0.01) decreased with their respective exposures. GPx activities decreased significantly (p < 0.05–0.01) in all the tissues with 3 h heat stress and increased in liver and kidney (p < 0.05–0.01) with 48 h deltamethrin stress when compared to control group. Heat stressed fish exposed to deltamethrin exhibited significantly (p < 0.05–0.01) lower and higher activities respectively in liver and kidney as compared to controls along

Table 5 Enzymatic antioxidants as modulated by heat stress and deltamethrin in the liver of Channa punctata Bloch. Groups

Control Heat Stress (HS) Deltamethrin (DEL) HS + DEL DEL + HS

Parameters Catalase (nmol H2O2 consumed/min/mg protein)

Glutathione S-transferase (nmol CDNB conjugates/min/mg protein)

Glutathione peroxidase (nmol NADPH oxidised/min/mg protein)

Glutathione reductase (nmol NADPH oxidized/min/mg protein)

112.5 ± 10.2 159.6 ± 12.6a

298.5 ± 4.6 238.2 ± 14.3a

241.8 ± 4.0 175.7 ± 11.6b

203.8 ± 13.0 163.9 ± 7.2a

69.2 ± 5.3a

357.1 ± 2.6b,q

328.1 ± 12.5b

349.0 ± 16.7b

95.7 ± 3.1q,x 34.3 ± 1.8b,q,y

276.3 ± 26.9x 337.3 ± 4.2b,q,x

201.5 ± 10.2a,y 281.6 ± 6.4b,q,x

240.9 ± 11.8a,q,y 305.1 ± 7.1b,q,x

Values are expressed as means ± SE, n = 5. Significant difference is shown as ap < 0.05, bp < 0.01 when compared with controls; pp < 0.05, qp < 0.01 when compared with heat stressed (3 h) group and xp < 0.05, yp < 0.01 when compared with deltamethrin (0.75 ppb  48 h) only exposed group.

Table 6 Activities of enzymatic antioxidants as modulated by heat stress and deltamethrin in the kidney of Channa punctata Bloch. Groups

Control Heat Stress (HS) Deltamethrin (DEL) HS + DEL DEL + HS

Parameters Catalase (nmol H2O2 consumed/min/mg protein)

Glutathione S-transferase (nmol CDNB conjugates/min/mg protein)

Glutathione peroxidase (nmol NADPH oxidised/min/mg protein)

Glutathione reductase (nmol NADPH oxidized/min/mg protein)

210.6 ± 8.2 159.3 ± 11.7a

314.3 ± 25.4 257.0 ± 13.2a

274.5 ± 18.1 170.9 ± 12.1b

220.8 ± 10.3 184.8 ± 7.6b

126.6 ± 14.4b

427.4 ± 15.6a

428.1 ± 15.7b

280.6 ± 10.7b

189.3 ± 7.0y 198.8 ± 6.4p,y

409.5 ± 23.1a,q 615.7 ± 81.9a,q,x

473.0 ± 12.2b,p,x 287.6 ± 43.3

336.1 ± 32.1a,y 293.0 ± 31.9a,p

Values are expressed as means ± SE, n = 5. Significant difference is shown as ap < 0.05, bp < 0.01 when compared with controls; pp < 0.05, qp < 0.01 when compared with heat stressed (3 h) group and xp < 0.05, yp < 0.01 when compared with deltamethrin (0.75 ppb  48 h) only exposed group.

Table 7 Activities of enzymatic antioxidants as modulated by heat stress and deltamethrin in the gills of Channa punctata Bloch. Groups

Control Heat Stress (HS) Deltamethrin (DEL) HS + DEL DEL + HS

Parameters Catalase (nmol H2O2 consumed/min/mg protein)

Glutathione S-transferase (nmol CDNB conjugates/min/mg protein)

Glutathione peroxidase (nmol NADPH oxidised/min/mg protein)

Glutathione reductase (nmol NADPH oxidized/min/mg protein)

211.7 ± 11.0 140.4 ± 7.7b

288.4 ± 3.0 199.6 ± 5.2b

392.6 ± 17.4 304.7 ± 16.9b

242.9 ± 11.5 212.9 ± 14.8b

153.4 ± 9.7b

221.3 ± 15.8b

293.5 ± 14.9b

340.1 ± 15.2b

186.6 ± 4.4b,q,x 152.8 ± 2.9b

205.2 ± 9.8b 164.6 ± 9.0b,p,x

277.6 ± 20.8b,q 261.9 ± 46.9b

273.6 ± 15.2b,q,y 342.3 ± 9.2b,q

Values are expressed as means ± SE, n = 5. Significant difference is shown as bp < 0.01 when compared with controls; pp < 0.05, qp < 0.01 when compared with heat stressed (3 h) group and xp < 0.05, yp < 0.01 when compared with deltamethrin (0.75 ppb  48 h) only exposed group.

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with single exposure groups. When deltamethrin exposed fish were subsequently given heat stress the activity of GPx was significantly (p < 0.05–0.01) higher as compared to heat stressed group and lower than deltamethrin-exposed group. In gills, all the exposures significantly decreased (p < 0.05–0.01) GPx activity (Tables 5–7). GR followed the pattern of GPx in liver and kidney, but in gills the response was different. The GR activity was significantly (p < 0.01) higher in deltamethrin-exposed group and in the dual exposure groups as compared to controls. The activities in the groups of combined exposures were significantly (p < 0.01) higher, when compared with control, than their respective exposures (Tables 5–7).

4. Discussion In the present study, deltamethrin-induced stress response in C. punctata, was manifested as induction of HSP70 and alterations of various biochemical parameters. Additionally, heat stress augmented toxic effects of deltamethrin on certain biochemical parameters, more importantly the antioxidants in most of the tissues. Deltamethrin and other pyrethroids possess a low degree of toxicity and are considered to be environmentally less persistent [38]. Since their introduction the toxicity of pyrethroids has been widely studied in different animal models including fish. Pyrethroids have been shown to be lethal to fish at concentrations 10–1000 times lower than corresponding values for mammals and birds [39,40]. An interactive effect of contaminants has been reported in fish [41,42]. In the present study with deltamethrin and elevated temperature as stress, the combined exposure revealed differential results in all the parameters studied when compared to their individual respective exposures. According to Sekine et al. [43] increasing or decreasing temperature can influence toxicity of pollutants. The synergistic effect of water temperature and herbicide has been reported. Tarja et al. [44] observed significant decrease in activities of ethoxyresorufin-O-deethylase (EROD) with increasing water temperature in Oncorhynchus mykiss. Therefore, the presence of pyrethroids in the aquatic environment and water temperature may modulate the toxic impact on fish [45]. In addition to heat stress, deltamethrin at the concentration of 0.75 ppb for 48 h induced HSP70 in different tissues. A noticeable amount of constitutive HSP70 was also found in all the tissues. Yu et al. [46] and Fader et al. [47] have demonstrated that fish and other aquatic organisms maintain detectable concentrations of constitutive HSP70. The combined exposure of both stressors showed synergistic response in the induction of HSP70. In liver, where heat stress followed the deltamethrin exposure, HSP70 values were lower than that induced by individual stressors. Levels of HSP in kidney were significantly lower in the group where heat stress followed the deltamethrin exposure when compared to deltamethrin exposed and heat stressed groups. In gills, HSP70 values in the heat stressed group were higher than deltamethrin exposed group as compared to controls and stressors in combination exhibited added response showing HSP levels higher than their individual responses. All these observations infer that conditions at the time of a pesticide challenge may modify the magnitude of response. The induction of hepatic HSP70 is reported to be different against various interacting abiotic factors in fish exposed to environmental pollutants [48]. The decrease in the levels of HSP70 could be supported by a study in brown trout (Salmo trutta f. fario) and stone loach (Barbatula barbatula) exposed to mixtures of environmental pollutants in laboratory, semi-field and in field studies. It was revealed that the stress response follows an optimum curve, resulting in a maximum HSP70 level under stress but rather low

HSP70 levels when stressors (chemicals, high temperature) become too severe. It has also been reported that the expression of HSP in fishes may be subjected to seasonal variation [47]. Acute toxicity of deltamethrin on various aspects in fish has been reported in Oreochromis niloticus L., Cyprinus carpio L. Carassius auratus gibelio Bloch, and C. Punctata, [11,12,14,49]. Toxicity of pyrethroids has also been reported in zooplankton communities, some beneficial aquatic arthropods, lobster and shrimp [50,51]. Besides the induction of HSP70, the stressors significantly increased the formation of protein carbonyls. Thus, protein oxidation appeared to be one of the apparent toxic effects of the stressors. When one stress followed the other, the response in the carbonylation of proteins was different than that observed with individual stressor. Protein carbonyl formation can occur as a result of oxidative stress. An increase in the levels of carbonyl groups correlates well with protein damage caused by oxidative stress [52]. The induction of protein carbonyl in fish was identified as a potentially useful biomarker of oxidative damage in C. punctata and Zoarces viviparus [53,54]. The level of protein carbonyl was consistent with the induction of HSP70. When subjected to treatments causing protein damage (proteotoxicity) stress-proteins are upregulated proportionately to the degree of stress [55]. Under these adverse conditions stress-proteins are thought to counter proteotoxic effects [56]. Exposure to deltamethrin and 3 h heat stress induced a perceptible oxidative stress in fish. Oxidative stress-inducing effect of deltamethrin has been established in fish [10]. Deltamethrin and heat stress significantly increased peroxidation of lipids in all the tissues Braguini et al. [8] attributed the toxicity of deltamethrin to perturbations in lipid–lipid, lipid–protein interactions and interference in transport mechanisms. Heat stress has also been reported to cause lipid peroxidation [57,58] which is also evident with the findings in the present study. In liver and kidney, extent of LPO in the combined exposure of deltamethrin and heat stress showed lower values than the two exposures individually. But in gills, the differential response of single and combined exposures was noteworthy where combined exposures of deltamethrin and heat stress did not exhibit any additional effect in toxicity. The responses of gills may be attributed to the high levels of HSP70 induction following combined exposures. Su et al. [59] have reported a similar observation of decreased LPO values involving HSP70. Thiols, including GSH are also an integral part of antioxidant system [60]. In response to stress, thiols are reported to be modulated by the cells, as they are the first to be used in cellular defense against stress [61]. Fish responded to deltamethrin exposure with increased GSH but the response to heat shock for 3 h was opposite to that of deltamethrin where GSH decreased (Fig. 4). In group of fish which was exposed to deltamethrin after heat stress, no significant increase was observed in GSH content. However, fish pre-exposed to deltamethrin followed with heat showed increase in GSH content in the liver and decrease in gills. Even when deltamethrin was given after heat stress it was unable to lift the GSH content of heat stress whereas heat stress to deltamethrin pre-exposed further increased the GSH in liver but decrease the same in gills showing that they are more vulnerable than the liver. Increase in GSH content has been described as one of the protective mechanisms that fish adopt in the initial phases of exposure to aquatic pollutants [62,63]. Thiol profile was also modulated by deltamethrin and heat stress in combination or in their individual exposure. Combined stress of deltamethrin and heat one after the other showed differential response in different tissues. Increase in the activities of antioxidants has been reported to be a general response of fish when exposed to environmental contaminants [63]. To neutralize the impact of reactive oxygen species (ROS), both enzymatic and non-enzymatic antioxidants are activated [64]. Similarly, deltamethrin exposure results in significant

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increase in activities of glutathione-dependent antioxidant enzymes. A significant increase was recorded in the activities of GST, GR and GPx in liver and kidney, while there was a significant decrease in the activities of GST and GPx in gills as in heat stress where all glutathione-dependent antioxidant enzymes were reported to decrease in gills besides in liver and kidney. The decrease in the activities of antioxidant enzymes may result when the stress is overwhelmed and cannot be compensated anymore [65]. The exposure to deltamethrin and heat stress caused a significant decrease in the activity of catalase in all the organs. This decrease in catalase activity could be attributed to the fluctuation of superoxide radicals, which have been accounted to impede CAT activity [66]. In deltamethrin exposed fish, the levels of GPx, GST and GSH were found elevated, apparently to provide protection against ROS damage. Antioxidant responses and oxidative stress have been used as biomarkers of exposure [22,67]. A high rate of absorption of deltamethrin through gills also makes fish vulnerable to its toxicity [50]. Once in the aquatic environment, pesticides can have deleterious effects on aquatic organisms. Exposure to pesticides in combination with other agents may exert effects different from those experienced with pesticides alone. As the physiology of fish is strongly related to temperature due to its ambient environment, their response to chemical exposures is also influenced by temperature. Although reports are available on the extent of oxidative damage and antioxidant mechanisms in fish on exposure to deltamethrin, no such attempt has so far been made on the aspect regarding its response under other stressors, environmental or otherwise including heat stress. Seasonal temperature changes have profound effects on the physiology of ectotherms, resulting in altered toxicity of chemicals. In the present study, we have demonstrated that modulation in one of the environmental variable such as temperature has significant effect on toxicity outcome of pesticide deltamethrin. This study has an ecological relevance from the view point of effect of temperature on aquatic organisms. It also provides an insight into the adaptation and survival of a fish community in a habitat where temperature elevation is quite frequent in concurrence with exposure to environmental toxicants. Conflict of interest statement Authors declare that there are no conflicts of interest. Acknowledgements Financial support of Council of Scientific and Industrial Research (CSIR) Government of India to Manpreet Kaur in the form of Research Associateship is gratefully acknowledged. References [1] S. Peuranen, M. Keinanen, C. Tigerstedt, P.J. Vuorinen, Effects of temperature on the recovery of juvenile grayling (Thymallus thymallus) from exposure to Al + Fe, Aquat. Toxicol. 65 (2003) 73–84. [2] T.D. Clark, E. Sandblom, G.K. Cox, S.G. Hinch, A.P. Farrell, Circulatory limits to oxygen supply during an acute temperature increase in the Chinook salmon (Oncorhynchus tshawytscha), Am. J. Physiol. Regul. Integr. Comp. Physiol. 295 (2008) R1631–R1639. [3] J.C. Perez-Casanova, M.L. Rise, B. Dixon, L.O. Afonso, J.R. Hall, S.C. Johnson, A.K. Gamperl, The immune and stress responses of Atlantic cod to long-term increases in water temperature, Fish Shellfish Immunol. 24 (2008) 600–609. [4] R.S. Hattori, J.I. Fernandino, A. Kishii, H. Kimura, T. Kinno, M. Oura, G.M. Somoza, M. Yokota, C.A. Strussmann, S. Watanabe, Cortisol-induced masculinization: does thermal stress affect gonadal fate in pejerrey, a teleost fish with temperature-dependent sex determination? PLoS One 4 (2009) e6548. [5] K. Said Ali, A. Ferencz, J. Nemcsok, E. Hermesz, Expressions of heat shock and metallothionein genes in the heart of common carp (Cyprinus carpio): effects of temperature shock and heavy metal exposure, Acta Biol. Hung. 61 (2010) 10– 23.

223

[6] T. Smital, T. Luckenbach, R. Sauerborn, A.M. Hamdoun, R.L. Vega, D. Epel, Emerging contaminants – pesticides, PPCPs, microbial degradation products and natural substances as inhibitors of multixenobiotic defense in aquatic organisms, Mutat. Res. 552 (2004) 101–117. [7] A. Slaninova, M. Smutna, H. Modra, Z. Svobodova, A review: oxidative stress in fish induced by pesticides, Neuro. Endocrinol. Lett. 30 (Suppl. 1) (2009) 2–12. [8] W.L. Braguini, S.M. Cadena, E.G. Carnieri, M.E. Rocha, M.B. de Oliveira, Effects of deltamethrin on functions of rat liver mitochondria and on native and synthetic model membranes, Toxicol. Lett. 152 (2004) 191–202. [9] I. Sayeed, S. Parvez, S. Pandey, B. Bin-Hafeez, R. Haque, S. Raisuddin, Oxidative stress biomarkers of exposure to deltamethrin in freshwater fish, Channa punctatus Bloch, Ecotoxicol. Environ. Saf. 56 (2003) 295–301. [10] F. Atif, S. Parvez, S. Pandey, M. Ali, M. Kaur, H. Rehman, H.A. Khan, S. Raisuddin, Modulatory effect of cadmium exposure on deltamethrin-induced oxidative stress in Channa punctata Bloch, Arch. Environ. Contam. Toxicol. 49 (2005) 371–377. [11] M.Z. Yildirim, A.C. Benli, M. Selvi, A. Ozkul, F. Erkoc, O. Kocak, Acute toxicity, behavioral changes, and histopathological effects of deltamethrin on tissues (gills, liver, brain, spleen, kidney, muscle, skin) of Nile tilapia (Oreochromis niloticus L.) fingerlings, Environ. Toxicol. 21 (2006) 614–620. [12] J. Velisek, R. Dobsikova, Z. Svobodova, H. Modra, V. Luskova, Effect of deltamethrin on the biochemical profile of common carp (Cyprinus carpio L.), Bull. Environ. Contam. Toxicol. 76 (2006) 992–998. [13] M.A. Ansari, R.K. Razdan, Concurrent control of mosquitoes and domestic pests by use of deltamethrin-treated curtains in the New Delhi Municipal Committee, India, J. Am. Mosquito Control Assoc. 17 (2001) 131–136. [14] D. Dinu, D. Marinescu, M.C. Munteanu, A.C. Staicu, M. Costache, A. Dinischiotu, Modulatory effects of deltamethrin on antioxidant defense mechanisms and lipid peroxidation in Carassius auratus gibelio liver and intestine, Arch. Environ. Contam. Toxicol. 58 (2010) 757–764. [15] A. Hegyi, T. Beres, L. Varadi, K.K. Lefler, B. Toth, B. Urbanyi, Investigation of long-term stress induced by several stressors by determination of the concentration of different blood plasma components in a model of Prussian carp (Carassius auratus gibelio BLOCH, and Common carp (Cyprinus carpio L., 1758), Acta Biol. Hung. 57 (2006) (1783) 301–313. [16] C.B. Schreck, Accumulation and long-term effects of stress in fish, in: G.P. Moberg, J.A. Mench (Eds.), The Biology of Animal Stress, CABI Publishing, Wallingford, UK, 2002, pp. 147–158. [17] A. Kaetsu, T. Fukushima, S. Inoue, H. Lim, M. Moriyama, Role of heat shock protein 60 (HSP60) on paraquat intoxication, J. Appl. Toxicol. 21 (2001) 425– 430. [18] M.J. Snyder, E.P. Mulder, Environmental endocrine disruption in decapod crustacean larvae, hormone titers, cytochrome P450, and stress protein responses to heptachlor exposure, Aquat. Toxicol. 55 (2001) 177–190. [19] S.B. Ceyhun, M. Senturk, D. Ekinci, O. Erdogan, A. Ciltas, E.M. Kocaman, Deltamethrin attenuates antioxidant defense system and induces the expression of heat shock protein 70 in rainbow trout, Comp. Biochem. Physiol. C Toxicol. Pharmacol. 152 (2010) 215–223. [20] H.N. Yang, H.C. Chen, Uptake and elimination of cadmium by Japanese eel, Anguilla japonica, at various temperatures, Bull. Environ. Contam. Toxicol. 56 (1996) 670–676. [21] M. Marinovich, F. Ghilardi, C.L. Galli, Effect of pesticide mixtures on in vitro nervous cells, comparison with single pesticides, Toxicology 108 (1996) 201– 206. [22] A. Valavanidis, T. Vlahogianni, M. Dassenakis, M. Scoullos, Molecular biomarkers of oxidative stress in aquatic organisms in relation to toxic environmental pollutants, Ecotoxicol. Environ. Saf. 64 (2006) 178–189 (review). [23] K. Wirtz, S. Bala, A. Amann, A. Elbert, A promise extended – future role of pyrethroids in agriculture, Bayer Crop Sci. J. 62 (2009) 145–154. [24] L.S. Clesceri, A.E. Greenberg, A.D. Eaton, Standard Methods for the Examination of Water and Wastewater, 20th ed., American Public Health Association (APHA), Washington, DC, 1998. [25] R.L. Anderson, C.Y. Wang, I.V. Kersen, K.J. Lee, W.J. Welch, P. Lavagnini, G.M. Hahn, An immunoassay for heat shock protein 73/72, use of the assay to correlate HSP73/72 levels in mammalian cells with heat response, Int. J. Hyperther. 9 (1993) 539–552. [26] B.S. Washburn, J.J. Moreland, A.M. Slaughter, I. Werner, D.E. Hinton, B.M. Sanders, Effects of handling on heat shock protein expression in rainbow trout (Oncorhynchus mykiss), Environ. Toxicol. Chem. 2 (2002) 557–560. [27] U.K. Laemmli, Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature 227 (1970) 680–685. [28] E. Floor, G. Wetzel, Increased protein oxidation in human substantial nigra pars compacta in comparison with basal ganglia and prefrontal cortex measured with an improved dinitrophenyl hydrazine, J. Neurochem. 70 (1998) 268–275. [29] M. Mihara, M. Uchiyama, Determination of malonaldehyde precursor in tissues by thiobarbituric acid test, Anal. Biochem. 86 (1978) 271–278. [30] D.W. Jollow, J.R. Mitchell, N. Zampagilone, J.R. Gilete, Bromobenzene induced liver necrosis, protective role of glutathione and evidence for 3,4bromobenzeneoxide as a hepatotoxic intermediate, Pharmacology 11 (1974) 151–169. [31] J. Sedlak, H.R. Lindsay, Estimation of total, protein-bound and non-protein sulfhydryl groups in tissues with Ellman’s reagent, Anal. Biochem. 25 (1968) 192–205.

224

M. Kaur et al. / Chemico-Biological Interactions 193 (2011) 216–224

[32] S. Parvez, I. Sayeed, S. Pandey, A. Ahmad, B. Bin-Hafeez, R. Haque, I. Ahmad, S. Raisuddin, Modulatory effect of copper on non-enzymatic antioxidants in freshwater fish Channa punctatus (Bloch), Biol. Trace Elem. Res. 93 (2003) 237– 248. [33] A. Claiborne, Catalase activity, in: R.A. Greenwald (Ed.), CRC HandBook of Methods in Oxygen Radical Research, CRC Press Inc., Boca Raton, FL, 1985, pp. 283–284. [34] W.H. Habig, M.J. Pabst, W.B. Jokoby, Glutathione S-transferase: the first enzymatic step in mercapturic acid formation, J. Biol. Chem. 249 (1974) 7130– 7139. [35] M. Mohandas, J.J. Marshall, G.G. Duggin, J.S. Horvath, D. Tiller, Differential distribution of glutathione and glutathione related enzymes in rabbit kidney, Cancer Res. 44 (1984) 5086–5091. [36] S. Pandey, S. Parvez, I. Sayeed, R. Haque, B. Bin-Hafeez, S. Raisuddin, Biomarkers of oxidative stress: a comparative study of river Yamuna fish Wallago attu (Bl. & Schn.), Sci. Tot. Environ. 309 (2003) 105–115. [37] O.H. Lowry, N.J. Rosenbrough, A.L. Farr, R.J. Randall, Protein measurement with Folin phenol reagent, J. Biol. Chem. 193 (1951) 265–275. [38] M. Villarini, M. Moretti, R. Pasquini, G. Scassellati-Sforzolini, C. Fatigoni, M. Marcarelli, S. Monarca, A.V. Rodrıguez, In vitro genotoxic effects of the insecticide deltamethrin in human peripheral blood leukocytes: DNA damage (‘comet’ assay) in relation to the induction of sister-chromatid exchanges and micronuclei, Toxicology 130 (1998) 129–139. [39] S.P. Bradbury, J.R. Coats, Toxicological and toxicodynamics of pyrethroid insecticide in fish, Environ. Toxicol. Chem. 8 (1989) 373–386. [40] J.T. Eells, J.L. Rasmussen, P.A. Bandettini, J.M. Propp, Differences in the neuroexcitatory actions of pyrethroid insecticides and sodium channelspecific neurotoxins in rat and trout brain synaptosomes, Toxicol. Appl. Pharmacol. 123 (1993) 107–119. [41] M. Calta, M.S. Ural, Acute toxicity of the synthetic pyrethroid deltamethrin to young mirror carp, Cyprinus carpio, Fresen. Environ. Bull. 13 (2004) 1179– 1183. [42] R. Viran, F.U. Erkoc, H. Polat, O. Kocak, Investigation of acute toxicity of deltamethrin on guppies (Poecilia reticulata), Ecotoxicol. Environ. Saf. 55 (2003) 82–85. [43] M. Sekine, H. Nakanishi, M. Ukita, Study on fish mortality caused by the combined effects of pesticides and changes in environmental conditions, Ecol. Model. 86 (1996) 259–264. [44] N. Tarja, E. Kirsti, L. Marja, E. Kari, Thermal and metabolic factors affecting bioaccumulation of triazine herbicides by rainbow trout (Oncorhynchus mykiss), Environ. Toxicol. 18 (2003) 219–226. [45] A. Moore, C.P. Waring, The effects of a synthetic pyrethroid pesticide on some aspects of reproduction in Atlantic salmon (Salmo salar L.), Aquat. Toxicol. 52 (2001) 1–12. [46] Z. Yu, W.E. Magee, J.R. Spotila, Monoclonal antibody ELISA test indicates that large amounts of constitutive hsp-70 are present in salamanders, turtle and fish, J. Therm. Biol. 19 (1994) 41–53. [47] S.C. Fader, Z.M. Yu, J.R. Spotila, Seasonal variation in heat shock proteins (hsp70) in stream fish under natural conditions, J. Ther. Biol. 19 (1994) 335– 341. [48] H.R. Kohler, C. Bartussek, H. Eckwert, K. Farian, S. Granzer, T. Knigge, N. Kunz, The hepatic stress protein (hsp70) response to interacting abiotic parameters in fish exposed to various levels of pollution, J. Aquat. Ecosys. Stress Recov. 8 (2001) 261–279. [49] R.A. Ansari, M. Kaur, F. Ahmad, S. Rahman, H. Rashid, F. Islam, S. Raisuddin, Genotoxic and oxidative stress-inducing effects of deltamethrin in the

[50]

[51]

[52] [53]

[54]

[55]

[56] [57]

[58]

[59]

[60] [61] [62]

[63]

[64]

[65]

[66] [67]

erythrocytes of a freshwater biomarker fish species, Channa punctata Bloch, Environ. Toxicol. 24 (2009) 429–436. A.K. Srivastav, S.K. Srivastava, S.K. Srivastav, Impact of deltamethrin on serum calcium and inorganic phosphate of freshwater catsh, Heteropneustes fossilis, Bull. Environ. Contam. Toxicol. 59 (1997) 841–846. U. Friberg-Jensen, L. Wendt-Rasch, P. Woin, K. Christoffersen, Effects of the pyrethroid insecticide, cypermethrin, on a freshwater community studied under field conditions I. Direct and indirect effects on abundance measures of organisms at different trophic levels, Aquat. Toxicol. 63 (2003) 357–371. I. Dalle-Donne, R. Rossi, D. Giustarini, A. Milzani, R. Colombo, Protein carbonyl groups as biomarkers of oxidative stress, Clin. Chim. Acta 329 (2003) 23–38. S. Parvez, S. Raisuddin, Protein carbonyls, novel biomarkers of exposure to oxidative stress-inducing pesticides in freshwater fish Channa punctata (Bloch), Environ. Toxicol. Pharmacol. 20 (2005) 112–117. B.C. Almroth, J. Sturve, A. Berglund, L. Forlin, Oxidative damage in eelpout (Zoarces viviparus), measured as protein carbonyls and TBARS, as biomarkers, Aquat. Toxicol. 73 (2005) 171–180. V. Dowling, P.C. Hoarau, M. Romeo, J. O’Halloran, F. van Pelt, N. O’Brien, D. Sheehan, Protein carbonylation and heat shock response in Ruditapes decussatus following p,p0 -dichlorodiphenyldichloroethylene (DDE) exposure: a proteomic approach reveals that DDE causes oxidative stress, Aquat. Toxicol. 77 (2005) 11–18. L.E. Hightower, Heat shock, stress proteins, chaperones, and proteotoxicity, Cell 66 (1991) 191–197. M.S. Parihar, T. Javeri, T. Hemnani, A.K. Dubey, P. Prakash, Responses of superoxide dismutase, glutathione peroxidase and reduced glutathione antioxidant defenses in gills of the fresh water catfish (Heteropneustes fossilis) to short-term elevated temperature, J. Therm. Biol. 22 (1997) 151–156. L.T. Chien, D.F. Hwang, Effects of thermal stress and vitamin C on lipid peroxidation and fatty acid composition in the liver of thornfish Terapon jarbua, Comp. Biochem. Physiol. B 128 (2001) 91–97. C.Y. Su, K.Y. Chong, K. Edelstein, S. Lille, R. Khardori, C.C. Lai, Constitutive HSP70 attenuates hydrogen peroxide-induced membrane lipid peroxidation, Biochem. Biophys. Res. Commun. 265 (1999) 279–284. A. Pastore, G. Federici, E. Bertini, F. Piemonte, Analysis of glutathione: implication in redox and detoxification, Clin. Chim. Acta 333 (2003) 19–39. D.A. Dickinson, H.J. Forman, Glutathione in defense and signaling, lessons from a small thiol, Ann. NY Acad. Sci. 973 (2002) 488–504. B.M. Hasspieler, J.V. Behar, R.T. Di Giulio, Glutathione-dependent defense in channel catfish (Ictalurus punctatus) and brown bullhead (Ameriurus nebulosus), Ecotoxicol. Environ. Saf. 28 (1994) 82–90. E. Stephensen, J. Sturve, L. Forlin, Effects of redox cycling compounds on glutathione content and activity of glutathione-related enzymes in rainbow trout liver, Comp. Biochem. Physiol. C 133 (2002) 435–442. M. Lopez-Torres, R. Perez-Campo, S. Cadenas, C. Rojas, G. Barja, A comparative study of free radicals in vertebrates – II. Non-enzymatic antioxidants and oxidative stress, Comp. Biochem. Physiol. B 105 (1993) 757–763. M. Kaur, F. Atif, M. Ali, H. Rehman, S. Raisuddin, Heat stress-induced alterations of antioxidants in the freshwater fish Channa punctata Bloch, J. Fish Biol. 67 (2005) 1653–1665. D.W. Filho, Fish antioxidant defenses – a comparative approach, Braz. J. Med. Biol. Res. 29 (1996) 1735–1742. L. Vigano, A. Arillo, C. Falugi, F. Melodia, S. Polesello, Biomarkers of exposure and effect in flounder (Platichthys flesus) exposed to sediments of the Adriatic sea, Mar. Poll. Bull. 42 (2001) 887–894.