Some hematological,biochemical, and enzymological parameters of a fresh-water teleost fish, Channa punctatus, exposed to sublethal concentrations of quinalphos

Alterations in the levels of hemoglobin, total plasma proteins, glucose, and lactic acid in the blood; glycogen and lactic acid content of liver and white skeletal muscle; and the activities of lactate dehydrogenase, pyruvate dehydrogenase, and succinate dehydrogenase in liver, kidney, intestine, brain, gills, and muscles were examined in the fresh-water snake-head fish, Channa punctatus, after exposure to a sublethal concentration (25 μg/liter) of quinalphos for 60 and 120 days. Hemoglobin, plasma protein, glucose, and lactic acid decreased in pesticide-exposed fish. The glycogen content of the liver and muscles increased but lactic acid decreased. Lactate dehydrogenase activity decreased in all six tissues. Pyruvate dehydrogenase activity of liver, kidney, gill, and muscle decreased, but the enzyme activity was elevated in intestine and brain. In intestine, succinate dehydrogenase activity was elevated, and in the remaining five tissues the enzyme activity was significantly reduced. The present study showed that formation of glycogen and its breakdown was impaired in the liver, and aerobic oxidation of nutrients was adversely affected in quinalphos-exposed fish.     

Effects of Melia azedarach on nutritional physiology and enzyme activities of the rice leaffolder Cnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae)

Laboratory assays were done to evaluate the effect of Melia azedarach L. (Rutales: Meliaceae) seed extract on nutritional indices and gut enzymes acid phosphatases, alkaline phosphatases, adenosinetriphosphatases, and lactate dehydrogenase of the rice leaffolder (RLF) Cnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae). Larvae were fed a treated rice-leaf diet containing the seed extract and their midgut was used for enzyme determination. Laboratory experiments showed that the seed extract suppressed the larval activity of C. medinalis even at a low dose. Gross dietary utilization (efficiency of conversion of ingested and digested food) of RLF decreased after ingesting the treated rice-leaf diet. Food consumption, digestion, relative consumption rate, efficiency of conversion of ingested food, efficiency of conversion of digested food, and relative growth rate values declined significantly. As compared to the control, consumption of the extract containing rice-leaf diet resulted in a 69% reduction of the acid phosphatases activity, a 71% reduction of the alkaline phosphatases activity, a 46% reduction of the adenosine triphosphatases activity, and a 52% inhibition of the lactate dehydrogenase activity.     

Effect of sublethal concentration of metasystox on selected oxidative enzymes,tissue respiration,and hematology of the freshwater air-breathing fish,Channa striatus (Bleeker)

Sublethal exposure (LC₅₀, 1.7 mg/liter) of metasystox decreased succinic dehydrogenase activity and tissue respiration, while lactic dehydrogenase activity increased in gill, brain, muscle, liver, and kidney tissues of Channa striatus (Bleeker). Metasystox also decreased blood parameters like WBC, MCV, MCH, and MCHC. Increase observed in RBC, PCV, and Hb was statistically significant. Possible reasons for these changes are discussed.     

Comparative studies on carbofuran-induced changes in some cytoplasmic and mitochondrial enzymes and proteins of epigeic, anecic and endogeic earthworms

Exposure of epigeic (Perionyx sansibaricus), anecic (Lampito mauritii) and endogeic (Metaphire posthuma) earthworms to four different concentrations (10 ppm, 20 ppm, 40 ppm and 80 ppm) of carbofuran for 16 days induced specific activities of cytoplasmic malate dehydrogenase (cMDH), mitochondrial malate dehydrogenase (mMDH) and lactate dehydrogenase (LDH). P. sansibaricus showed 33-52% increase in specific activities of aerobic (cMDH, mMDH) and anaerobic (LDH) enzymes and proteins (cytoplasmic and mitochondrial). In L. mauritii, the enhancement in enzyme and protein level was 30-46%. Similarly, M. posthuma indicated 29-43% increase in profile of enzymes and proteins. The significant increase was observed on 2nd or 3rd day and it was maximum on 16th day of carbofuran exposure. Maximum effect of carbofuran was on epigeic earthworm. The sensitivity of mitochondrial dehydrogenase and protein of different earthworm species to carbofuran appears to be associated with oxygen availability in different strata of soil habitats. Inductions in enzyme specific activity and protein content were concentration and time-dependent. The effect was more pronounced on aerobic enzymes (cMDH, mMDH) as compared to anaerobic (LDH) one. The increases in mitochondrial enzyme (47%) and protein (43%) were more than the cytoplasmic enzymes (35%) and protein (31%), which suggest a greater effect of carbofuran on respiratory metabolism of earthworms. The carbofuran-dependent increase in cytoplasmic and mitochondrial enzymes and proteins might be due to increased synthesis of metabolic enzymes and stress proteins.     

Influence of Atrazine and Roundup pesticides on biochemical and molecular aspects of Biomphalaria alexandrina snails

The excessive use of pesticides in agriculture has sparkled the interest of scientists in investigating the harmful effects of these compounds. The present study evaluates the pesticides Atrazine and Roundup (glyphosate) on biochemical and molecular aspects of Biomphalaria alexandrina snails. The results showed that LC₁₀ of these two pesticides caused considerable reduction in survival rates and egg production of treated snails. Additionally, Atrazine proved to be more toxic to B. alexandrina snails than Roundup. In treated snails, glucose concentration (GL) in the hemolymph as well as lactate (LT) and free amino acid (FAA) in soft tissues of treated snails increased while glycogen (GN), pyruvate (PV), total protein (TP), nucleic acids (DNA and RNA) levels in snail’s tissues decreased. The activities of glycogen phosphorylase (GP), superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR), succinic dehydrogenase (SDH), acetylcholinesterase (AChE), lactic dehydrogenase (LDH) and phosphatases (ACP and ALP) enzymes in homogenate of snail’s tissues were reduced in response to the treatment with the two pesticides while lipid peroxide (LP) and transaminases (GOT and GPT) activity increased (P < 0.001). The changes in the number, position and intensity of DNA bands induced by pesticides may be attributed to the fact that pesticide can induce genotoxicity through DNA damage. It was concluded that the pollution of the aquatic environment by Atrazine and Roundup pesticides, would adversely affect the metabolism of the B. alexandrina snails, and have adverse effects on its reproduction.     

The mechanism of fungistatic action of sec-butylamine: II. The effect of sec-butylamine on pyruvate oxidation by mitochondria of Penicillium digitatum and on the pyruvate dehydrogenase complex

sec-Butylamine at 5 mM inhibited the oxidation of pyruvate by mitochondria isolated from hyphae of Penicillium digitalum, but had little effect on the oxidation of citrate, isocitrate, succinate, malate, acetyl-coenzyme A, or reduced nicotinamide adenine dinucleotide. sec-Butylamine did not interfere with oxidative phosphorylation, as evidenced by similar $\text{P}\text{O}$ ratios in treated and control mitochondria. The pyruvate dehydrogenase complex (EC 1.2.4.1) isolated from young hyphae of P. digitatum was inhibited strongly by 20 mM sec-butylamine, whereas other tricarboxylic acid cycle enzymes were only slightly affected at most. Inhibition of the pyruvate dehydrogenase complex by sec-butylamine was competitive with respect to pyruvate. The K_i for sec-butylamine in the reaction was 1.38 × 10^?2M, and the K_m for pyruvate was 2.28 × 10^?4M. These observations and other evidence derived from studies with intact hyphae support the hypothesis that the pyruvate dehydrogenase complex is the primary site of the fungistatic action of sec-butylamine.     

Biological and biochemical responses of Biomphalaria alexandrina to some extracts of the plants Solanum siniacum and Artemisia judaica L.

The molluscicidal activity of cold water, boiled water, methanol, ethanol, acetone and chloroform extracts of Solanum siniacum and Artemisia judaica L. plants against Biomphalaria alexandrina snails was carried out. The tests revealed plant’s ethanol extract was more toxic to the snails than the other tested extracts. Therefore, it was tested against snails’ fecundity (M_x), reproduction rate (R₀) and their infection with Schistosoma mansoni miracidia. In addition, biochemical parameters and the activities of some enzymes in tissues of snails treated with the two tested plants were determined. As well, glucose concentration in snails’ hemolymph was evaluated. Exposure of B. alexandrina snails to plant’s ethanol extract led to a significant reduction in their survival and snails’ fecundity, reproduction rate. In addition, it caused a considerable reduction in the infectivity of S. mansoni miracidia to the snails. Also, it caused a reduction in number of cercariae per snail during the patent period and in the period of cercarial shedding. The results revealed that the glucose concentration in hemolymph and Lactate level in soft tissues of treated snails were increased (P < 0.001) while glycogen, total protein, the lipid content and the pyruvate level in snail’s tissues decreased (P < 0.001). The activities of lactic dehydrogenase (LDH), pyruvatekinase (PK) and cytochrome oxidase (CY) enzymes in homogenate of snail’s tissues were reduced (P < 0.001) in response to treatment with the two tested plants while protease (PR) activity increased (P < 0.001). It is concluded that the application of LC₂₅ of ethanol extract of S. siniacum and A. judaica L. may be helpful in snail control as it interferes with the snails’ biology and physiology.     

Inhibition of permethrin-hydrolyzing esterases from Wiseana cervinata larvae

Inhibition of permethrin-hydrolyzing enzymes from larvae of the porina moth Wiseana cervinata has been examined in vivo and in vitro. Significant inhibition was shown by carbaryl and pirimiphos-methyl. 1-Dodecylimidazole substantially inhibited permethrin hydrolysis only in liver insects. The triphenylmethane dye tetrabromophenolphthalein was a moderate inhibitor only in vitro. TMDM (bis(N-dimethyl-4-aminophenyl)methane) had little effect on hydrolysis. These observations extend the range of species and substrates for which the triphenylmethane dyes and 1-dodecylimidazole are useful inhibitors of insecticide metabolism.     

Glucose metabolism in hepatopancreas and gill of Lamellidens marginallis during methyl parathion toxicity

Changes in glucose metabolism were studied in hepatopancreas and gill of freshwater mussel, Lamellidens marginalis, exposed to a sublethal concentration (8 ppm) of methyl parathion. A slight decrease in glycogen and pyruvate and an increase in lactate levels were observed. An increase in phosphorylase and aldolase suggested increased formation of trioses during methyl parathion toxicity. The decrease in lactate dehydrogenase activity and increase in lactate content indicated reduced mobilization of pyruvate into the citric acid cycle. Glucose-6-phosphate dehydrogenase activity was increased, suggesting enhanced oxidation of glucose by the HMP shunt pathway. Citric acid cycle enzymes such as isocitrate, succinate, and malate dehydrogenases were found to be decreased, suggesting abnormality in mitochondrial oxidative metabolism as a consequence of methyl parathion toxicity. The decreased cytochrome c oxidase and Mg²⁺-ATPase, apart from citric acid cycle enzymes, indicated impaired energy synthesis as a result of reduced aerobic oxidation of glycose. The increase in acid and alkaline phosphatase activities suggested enhanced breakdown of phosphate to release energy in view of inhibition of the ATPase system during methyl parathion stress. The changes were more pronounced in hepatopancreas as compared to gill of mussel exposed to methyl parathion.     

内生放线菌A-1对苹果果实轮纹病的防效及防御性酶活性的影响

为揭示内生放线菌A-1对苹果果实轮纹病的防效及防病机制,采用平板法和喷雾处理刺伤接种法,测定了其对苹果轮纹病菌的抑制作用及果实内防御性酶活性的影响。结果表明：菌株A-1能显著抑制轮纹病菌菌丝生长,10⁴ CFU/mL菌悬液的抑制率达90%以上;喷施10⁷ CFU/mL A-1菌悬液后,间隔12 h以上接种的各处理,3 d和7 d时防效分别为91.79%~95.67%和77.41%~94.00%,均与对照农药苯醚甲环唑相当。喷施10⁷ CFU/mL A-1菌悬液后接种或不接种轮纹病菌的处理,果实内过氧化物酶、过氧化氢酶、多酚氧化酶和苯丙氨酸解氨酶活性显著升高,且均高于只接种轮纹病菌的处理;喷施A-1菌悬液后接种病原菌的处理酶活性增加最显著,其峰值是对照的2.30~11.00倍。表明内生放线菌A-1可通过产生拮抗物质、提高寄主防御性酶活性等机制实现对苹果轮纹病的有效防治。     

群居型和散居型亚洲小车蝗成虫能量代谢相关酶活性测定

为明确亚洲小车蝗Oedaleus asiaticus成虫飞行肌对其体内能源物质利用的情况,采用室内生化法测定能源物质代谢相关5种酶,即3-磷酸甘油醛脱氢酶(glyceraldehyde phosphate dehydrogenase,GAPDH)、3-磷酸甘油脱氢酶(glycerol-3-phosphate dehyd...     

Differential influence of herbicide treatments on activity and kinetic parameters of C4 photosynthetic enzymes from Zea mays

The recommended field dose of rimsulfuron, imazethapyr, alachlor, atrazine or fluometuron differentially reduced shoot fresh and dry weight of 10-day-old maize seedlings as well as leaf protein content during the following 12 days. These reductions seemed consistent during the whole period with fluometuron, atrazine and alachlor but appeared to be nullified by the 5th day of treatment with rimsulfuron and imazethapyr. On the other hand, all herbicides mostly provoked significant inhibitions in specific activities of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31), malate dehydrogenase (MDH, EC 1.1.1.82), pyruvate phosphate dikinase (PPDK, EC 2.7.9.1) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39) in leaves during the first 2 days. Thereafter, the inhibition was recovered in samples treated with rimsulfuron and imazethapyr, leveled off with alachlor but consistently augmented with atrazine and fluometuron. The kinetic characterization showed that rimsulfuron or imazethapyr unchanged V_max of all enzymes in vitro, however, V_max of PEPC, PPDK and Rubisco were decreased in vivo. Nevertheless, atrazine or fluometuron substantially reduced V_max of all enzymes while alachlor showed a reduction in this value of PEPC, MDH and Rubisco. Thus atrazine, fluometuron and, to a lower extent, alachlor reduced concentrations of all enzymes as well as rimsulfuron and imazethapyr for only Rubisco. On the contrary, K_m values of all enzymes were progressively increased by all herbicides indicating that the different herbicides altered the structural integrity of all enzymes. These findings conclude that rimsulfuron or imazethapyr competitively inhibited MDH but revealed mixed inhibition to PEPC, PPDK and Rubisco. Atrazine or fluometuron revealed mixed inhibitions to all enzymes whereas alachlor seemed to be either a competitive inhibitor to PPDK or a mixed inhibitor to PEPC, MDH and Rubisco.     

Characterization of acetylcholinesterases of acaricide resistant and susceptible strains of the cattle tick Boophilus microplus (Can.): II. The substrate specificity and catalytic efficiency of the critical enzyme component

Concentrations and turnover numbers have been determined for the critical acetylcholinesterase component of the organophosphorus susceptible Yeerongpilly and resistant Biarra and Ridgelands strains. The lower specific activity for the substrate acetylthiocholine in the resistant strains was shown to be due to decreased catalytic efficiency and not to any change in the titer of the enzyme. Thioester analogs of acetylthiocholine, with additional methylene groups between the carbonyl carbon and the quaternary nitrogen of the molecule and in the alkyl moiety, have been used to derive K_m and V_max values for the enzymes from each strain. An hypothesis is proposed concerning the alteration of the active site that has occurred to produce the dissimilar aberrant enzymes of the resistant strains and the implications of this hypothesis for inhibitor design are discussed.     

Influence of malathion on the catalytic potential of glutamate dehydrogenase in sheep brain

Malathion stimulated sheep brain glutamate dehydrogenase. The maximal velocity (V_max) and Michaelis-Menten constant (K_m) showed alterations characteristic of nonessential activation. The activation energy required by the enzyme subjected to pesticidal stress was found to decrease, suggesting a decrement in energy barrier for the catalysis of the enzyme. The possible implications of malathion-stimulated glutamate dehydrogenase activity are discussed.     

Ecophysiological category based toxicological responses in metabolism of earthworms: Impact of a pyrethroidal insecticide

Effects of alphamethrin (0.04 ppm, 0.08 ppm, 0.16 ppm, 0.32 ppm) on some metabolic dehydrogenases and proteins for 1, 2, 3, 4, 8 and 16 days using three ecologically different earthworm species (Perionyx sansibaricus, Lampito mauritii and Metaphire posthuma) were studied. The first significant effect was on 2nd/3rd day and maximum response was achieved on 16th day of exposure of alphamethrin at all concentrations. Similarly, maximum effect was obtained at 0.32 ppm alphamethrin at different exposure periods. It showed a dose- and duration-dependent inhibitory effects of a pyrethroid on enzymes and proteins of earthworms. There were approximately 47%, 43% and 41% declines in specific activities of cytoplasmic malate dehydrogenase (cMDH) of P. sansibaricus, L. mauritii and M. posthuma, respectively, in response to 0.32 ppm exposure of alphamethrin for 16 days. In case of similar treatment, specific activities of mitochondrial malate dehydrogenase (mMDH) showed 57%, 51% and 45% reductions in epigeic (P. sansibaricus), anecic (L. mauritii) and endogeic (M. posthuma) earthworms, respectively. Exposure of alphamethrin (0.32 ppm) also decreased the specific activities of lactate dehydrogenase (LDH) by 47% (P. sansibaricus), 35% (L. mauritii) and 39% (M. posthuma) in different types of earthworms. The reductions in specific activities of a Krebs cycle enzyme (mMDH) were greater than that of a glycolytic enzyme (LDH). The protein contents declined significantly in the earthworms exposed to alphamethrin. The decreases of 41%, 36% and 30% were obtained in cytoplasmic proteins of P. sansibaricus, L. mauritii and M. posthuma, respectively. However, the mitochondrial proteins showed reductions of 45% in P. sansibaricus, 46% in L. mauritii and 38% in M. posthuma in response to alphamethrin (0.32 ppm) intoxication for 16 days. The decrease in the protein content reflected an inhibitory effect of alphamethrin on protein turnover. The most pronounced effect of alphamethrin was on metabolic enzymes and proteins of P. sansibaricus and least effect was found on M. posthuma. This clearly showed that surface dwelling species of earthworms (epigeic) are more vulnerable to toxic chemical than deep burrowing species (endogeic). Thus a possible ecophysiological link exists between the toxicological responses and ecological categories at metabolic level in tropical earthworms.     

Concentration related responses of chlorpyriphos in antioxidant, anaerobic and protein synthesizing machinery of the freshwater fish, Heteropneustes fossilis

Impacts of chlorpyriphos (2, 4, 6 ppm) on the profiles of antioxidant (catalase) and anaerobic (lactate dehydrogenase) enzymes and other macromolecular contents (DNA, RNA, protein) of various tissues of the freshwater fish, Heteropneustes fossilis were studied. Chlorpyriphos significantly decreased the specific activity of catalase (CAT) and lactate dehydrogenase (LDH) in the brain, liver, gill and skeletal muscle of the fish. The reduction in specific activity might be due to the binding of chlorpyriphos or its metabolites with the enzyme molecules or affecting the synthesis and/or degradation of the enzymes. Like enzymes, the DNA, RNA and protein contents decreased in the brain, liver, gill and skeletal muscle of the fish as a function of increase in chlorpyriphos concentrations (2-6 ppm). The chlorpyriphos-induced reduction in these biochemical constituents might be because of alteration in their turnover (synthesis/degradation) in different tissues. The maximum effects on CAT, LDH, DNA, RNA and protein were obtained in response to 6 ppm chlorpyriphos. The present findings suggest chlorpyriphos concentration related impairment in antioxidative, anaerobic and protein synthesizing capacity of the fish. Therefore, the use of higher concentrations of chlorpyriphos should be avoided to protect the health of economically important freshwater food fish.     

The effect of single, binary, and tertiary combination of few plant derived molluscicides alone or in combination with synergist on different enzymes in the nervous tissues of the freshwater snail Lymnaea (Radix) acuminata (Lamark)

The effect of single, binary, and tertiary combination of few plant derived molluscicides alone or in combination with synergist on different enzymes (acetylcholinesterase—AChE, lactic dehydrogenase—LDH, and acid/alkaline phosphatase—ACP/ALP in the nervous tissue of the freshwater snail Lymnaea acuminata were studied. Sublethal in vivo 24 h exposure to 40 and 80% of LC₅₀ of Azadirachta indica oil (AI), oleoresin of Zingiber officinale (OL), Cedrus deodara oil (CD), Allium sativum (AS), and Polianthes tuberosa (PT) bulb powder singly, their binary combination of AI + OL, AS + CD, AS + PT, CD + OL, CD + PT, OL + PT, and tertiary combination of these binary combinations with the synergist piperonyl butoxide (PB) or MGK-264 significantly altered the activity of these enzymes. Tertiary combination with PB or MGK was very effective. Combination of CD + PT + MGK was more effective against AChE whereas, the combination of CD + OL + PB, CD + AS + PB, and CD + PT + PB were more effective against LDH, ACP, and ALP, respectively.     

Effects of the molluscicidal agent GA-C13:0, a natural occurring ginkgolic acid, on snail mitochondria

Ginkgolic acids (GAs) from the leaves and sarcotesta of Ginkgo biloba L. represent a new kind of molluscicide agent. To date, the mechanism(s) for the observed molluscicidal activity remains largely unknown. Since GA-C_13:0 has effectively inhibited snail mobility, we examined the effects of the compound on mitochondrial function and gene expression as compared to niclosamide. Snail mitochondrial damage induced by GAs was tested using transmission electron microscopy (TEM) and gene-expression profiling of five mitochondrial enzymes using real-time PCR. GA-C_13:0 was found to have a pronounced effect on snail mitochondria with gross ultrastructural changes. In addition, GA-C_13:0 was also found to inhibit the gene expression of four mitochondrial enzymes including cytochrome c oxidase, adenosine triphosphate (ATP) synthase, cytochrome b and dihydronicotinamide adenine dinucleotide (NADH) dehydrogenase. In contrast, niclosamide did not show such effects on mitochondrial function and gene expression, suggesting that the molluscicidal activity of GA-C_13:0 and niclosamide differed. Our results imply that snail mitochondria are a potential target for the molluscicidal activity of ginkgolic acids.     

Effect of liver enzyme induction on paraoxon metabolism in the rat

Orally administered [1-¹⁴C]ethyl paraoxon, O,O-diethyl-O-p-nitrophenyl phosphate, is readily absorbed from the gastrointestinal tract of male albino rats. Radioactivity is essentially eliminated in 72 hr by excretion into urine and feces and by expiration as ¹⁴CO₂. Compounds with radioactivity in the urine are tentatively identified as diethyl phosphoric acid, desethyl paraoxon, ethanol, metabolites conjugated with amino acids, and paraoxon; the first compound is the predominant radioactive metabolite. Intraperitoneally injected phenobarbital, DDT, dieldrin, and endrin are inducers of microsomal enzymes that degrade paraoxon. The aryl phosphate-cleaving activity in vitro is not dependent on the addition of NADPH. O-Dealkylation of paraoxon is catalyzed by microsomal enzymes that require NADPH and oxygen and are inhibited by carbon monoxide. Microsomal enzymes from rats pretreated with enzyme inducers give an increased rate of O-dealkylation of paraoxon. Reduced glutathione has little or no effect on paraoxon degradation by either microsomal or soluble enzymes. Actinomycin D inhibits O-dealkylation of paraoxon in vivo, as indicated by reduction of ¹⁴CO₂ formation, and in vitro, as indicated by decreased activity of microsomal O-dealkylase. The role of microsomal mixed-function oxidases and NADPH-dependent O-dealkylase in the metabolism of organophosphorus insecticides is discussed.     

Persistent sub-lethal chlorine exposure elicits the temperature induced stress responses in Cyprinus carpio early fingerlings

Thermal effluents discharged through cooling systems of nuclear power plants often contain chlorine (used to control bio-fouling), which may affect the metabolic status of fishes. In order to evaluate the hypothesis, we tested the effect of high temperature and a persistent sub-lethal chlorine exposure on stress responses in Cyprinus carpio advanced fingerlings. Fishes were acclimated to four different temperatures (26, 31, 33, and 36 °C) and maintained for 30 days in two different groups. Subsequently, one of the groups was exposed to persistent chlorine (0.1 mg L₋₁) for another 28 days and was compared with their respective temperature controls (without chlorine exposure). Sub-lethal doses of pollutants and increasing temperatures with in the tolerance range may not always register any morphological changes Therefore, we studied organ specific biochemical pathways viz. aspartate amino transferase, alanine amino transferase (enzymes of protein metabolism) in liver and muscle; fructose 1,6 diphosphatase (gluconeogenic pathway), in liver; pyruvate kinase, malate dehydrogenase, and lactate dehydrogenase (glycolytic pathway) in muscle; glucose-6-phosphate dehydrogenase (pentose phosphate pathway) in liver; alkaline phosphatase (phosphorus metabolism) in intestine, liver, and muscle; acetylcholine esterase (neurotransmitting enzyme) in brain, and adenosine triphosphate (for membrane transport) in gills at two different acclimation periods (14 and 28 days). The results indicate that C. carpio fingerlings demonstrated metabolic readjustments with increasing temperatures, in order to cope with energy demand of the cell. However, exposure to chlorine at higher temperatures affected protein metabolism, gluconeogenic pathway and subsequently glycolytic pathway, leading to an energy-limited condition. In addition, alteration of membrane transport and neurotransmission might be an early indication of cellular damage. Overall results indicate that persistent sub-lethal chlorine exposure elicits temperature induced stress response in C. carpio early fingerlings.