Consecutive emamectin benzoate and deltamethrin treatments affect the expressions and activities of detoxification enzymes in the rainbow trout (Oncorhynchus mykiss) (original) (raw)
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Aquaculture, 2011
Caligus rogercresseyi is a sea louse that affects salmon farming industry throughout the Southern hemisphere. In Chile, oral delivery of emamectin benzoate (EMB) to salmon was the main treatment used; however, C. rogercresseyi became resistant to EMB. The aims of this study were determine the effect of EMB on proteins involved in metabolism and drug resistance in rainbow trout (Oncorhynchus mykiss) and C. rogercresseyi. Since no commercial antibodies are available for these salmon's proteins, the expression of trout cytochrome P450 CYP2K1, CYP2M1, CYP3A and CYP3A27, and the multidrug transporters Pgp and MRP1 were investigated by semi-quantitative duplex RT-PCR. Trout specimens infested with C. rogercresseyi were medicated with EMB by a standard seven-day oral treatment in salmon farms from southern Chile, and collected between 10 and 25 days after the completion of medication. The samples examined were liver, muscle, gill, middle kidney and intestine from control and EMB treated rainbow trout, and C. rogercresseyi's total homogenates. In trout tissues most CYP and MDR gene expressions were up-regulated by EMB treatment, being MRP1 mRNA in all tissues, but particularly in the kidney and intestine, and CYP2M1 mRNA from muscle and gill, the most augmented. CYP3A, CYP3A27, CYP2K1 and Pgp mRNAs were slightly affected in the most tissues analyzed. In C. rogercresseyi minimal up-regulation of the expression for most of mRNA analyzed was observed, except for CYP2M1 mRNA, decreasing its expression by half, and CYP3A27 doubling its expression levels. Nonetheless, CYP3A mRNA expression was not detected in this parasite. These results suggest that treatment with EMB in salmon regulates the transcriptional expression of proteins involved in metabolism, distribution and elimination of endobiotics and xenobiotics, such as hormones and drugs, and even could affect the pharmacokinetics of EMB in the same treatment.► We examined transcriptional effect of emamectin benzoate on rainbow trout and Caligus rogercresseyi. ► Emamectin benzoate is used as antiparasitic against Caligus rogercresseyi. ► Most CYP and MDR genes expression were up-regulated in trout tissues ► MRP1 mRNA was the most augmented. ► In Caligus rogercresseyi minimal up-regulation was observed.
Aquatic Toxicology, 2007
Retene (7-isopropyl-1-methyl phenanthrene) is a polycyclic aromatic hydrocarbon (PAH), that causes dioxin-like toxicity to early life stages of fish, including increased rates of mortality, developmental defects characterized as blue sac disease (BSD), and induction of CYP1A enzymes. This study determined whether toxicity is associated with retene, or with its metabolism by CYP1A enzymes to hydroxylated derivatives. Larval rainbow trout (Oncorhynchus mykiss) were co-exposed to four concentrations of waterborne retene and four concentrations of waterborne α-naphthoflavone (ANF), a compound that antagonizes CYP1A induction and inhibits oxygenation reactions. The prevalence of mortality and BSD increased in an exposure-dependent manner for larvae exposed to retene alone. Tissue concentrations of CYP1A protein and retene metabolites also increased, but no un-metabolized retene (i.e., the parent compound) was measurable. At low concentrations of ANF, toxicity increased dramatically, while tissue concentrations of polar hydroxylated metabolites of retene decreased, and concentrations of less polar metabolites, and of parent retene, increased. At the highest concentration of ANF, retene toxicity was eliminated, and parent retene was the predominant form in tissue; no concentration of ANF was toxic by itself. The inhibition of retene hydroxylation and toxicity by ANF suggests that toxicity was caused by specific retene metabolites, and not by parent retene. The potentiation of retene toxicity at low concentrations of ANF, and the antagonism at high concentrations is a unique, non-linear interaction based on modulating CYP1A enzyme activity and retene metabolism. It demonstrates that effects on fish of different complex mixtures of hydrocarbons may not be easily predicted.
Environmental Toxicology and Chemistry, 1993
Mirror carp were exposed to Rotterdam Harbor sediment, highly contaminated with polychlorodibenzo-p-dioxins (PCDDs), polychlorodibenzofurans (PCDFs), and polychlorobiphenyls (PCBs) (0.5 pg 2,3,7,8-tetrachlorodibenzo-p-dioxin [TCDD] equivalents per kilogram dry weight). In two additional separate experiments rainbow trout (Oncorhynchus mykiss) and mirror carp (Cyprinus carpio) received a single intraperitoneal injection of approximately 0.01,0.03, 0.06,0.3, 0.6, or 3.0 pg TCDD per kilogram body weight. In all three experiments induction of hepatic P450 1A was measured with immunochemical and enzymatic methods. The polyclonal antibodies anticod (Gadus morhua), anti-perch (Percafluviatilis), and anti-rainbow trout P450 1A all cross-reacted with the P450 1A orthologue of the carp and rainbow trout. In most cases high correlations were found between 7-ethoxyresorufin 0-deethylation (EROD) activity and cytochrome P450 1A protein contents, the latter measured by the enzyme-linked immunosorbent assay (ELISA) and protein blot methods. However, the correlations between EROD activity and P450 1A protein levels were higher within the separate sampling periods (i.e., 3,6, and 12 weeks after dosage) than with the total data set, especially in the dose-effect study with the rainbow trout. This was probably caused by a difference in time-dependent relationships between P450 1A protein content and EROD enzyme activity: 12 weeks after dosage the P450 1A protein was still increased, although EROD activity had returned to background level. In addition, there were higher correlations of the EROD activity and P450 1A protein content with total P450 content in rainbow trout and carp treated with a single dose of TCDD, than with total P450 content in carp exposed to contaminated sediment. In our study, the ELISA method appeared to be more useful than the protein blot technique, because the ELISA is faster and has higher reproducibility. In addition, in all our experiments EROD activity showed a higher induction than the P450 1A protein, indicating a higher sensitivity of the EROD assay. Our results strongly indicated that determination of the P450 1A protein content and EROD activity provides complementary information. Thus we recommended the use of both the ELISA and the EROD activity assay in order to understand the nature of P450 1A induction.
Fish Physiology and Biochemistry - FISH PHYSIOL BIOCHEM, 1998
The effects of the 4-quinolones, oxolinic acid and flumequine on hepatic microsomal cytochrome P450 monooxygenases in rainbow trout were examined during this study. Following antibiotic administration in the diet for 10 days at a representative commercial medicated feed concentration, fish were killed for assay at various periods up to 12 days following a return to normal diet. Ethoxyresorufin O-deethylase (EROD) and benzyloxyresorufin O-dealkylase (BROD) activities were significantly elevated as a result of antibiotic treatment. The effects of oxolinic acid were delayed and longer-lasting compared with flumequine. The effects of flumequine were detectable 4 days earlier than those of oxolinic acid and lasted for less than ten days after treatment cessation. In contrast, oxolinic acid effects were apparent even on day 12 of the recovery period. The results of O-dealkylation, isoform-selective inhibition, and immunoblotting showed that the effects of both oxolinic acid and flumequine...
Aquatic Toxicology, 1994
in rainbow trout (Oncorhynchus mykiss) and cod ( aadus morhua) to 2,3,7,8-tetrachlorodibenzo-p-dioxin (2, 3,7,8-TCDD) Abstract 2,3,7,8-Tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) was administered intragastrically twice with 4 days interval, to juvenile cod and rainbow trout, total dose 8 ~g/kg body weight. Fish were killed after 9 and 17 days and the effects on hepatic xenobiotic metabolizing enzymes were determined by examining aldrin epoxidase (AE), glutathione-S-transferase (GST) against 1chloro-2,4-dinitrobenzene (CDNB) and the cytochrome P450-dependent ethoxyresorufin-Odeethylase (EROD) activities, and by immunoquantitating cytochrome P4501A1 using an indirect enzyme-linked immunosorbent assay (ELISA). AE and GST activities were not induced. However, 2,3,7,8-TCDD significantly induced EROD activities in rainbow trout and cod to 1450% and 415%, respectively, of the corresponding controls 9 days after the first treatment. The increase in EROD activity was supported by induction of the main catalyst P4501A1 as revealed by ELISA analyses. The distribution pattern of 14C-labelled 2,3,7,8-TCDD was studied by whole-body autoradiograpby. The liver concentration of radiolabelled compound in cod exceeded that of the rainbow trout. However, the degree of hepatic enzyme induction did not correspond to the concentration of radiolabelled 2,3,7,8-TCDD in the liver. Despite of the substantially higher level of 2,3,7,8-TCDD in the liver of cod, the EROD induction was lower in this species compared to rainbow trout.
Marine Environmental Research, 1992
A BS TRA C T lmhwtion of hepatic cvtochrorne P450-dependent microsomal monooxygenase by xenohiotics is a well-established phenomenon in teleost fish. As in htborato O' mammals, fish possess multiple forms of cytochrome P450 that display overlapping substrate specificity. One such isoJbrm, C YPIA 1, which has been chined and sequenced from rainbow trout, has been shown to be orthologous to rat C YP I A l and, as in mammals, is inducible up to several hundred-fold by planar aromatic hydrocarbons, PCBs and dioxins. It has been suggested that induction of CYP1A 1 orthologues might provide a sensitive biomonitor fi~r environmental pollution by mixtures of such compounds. In the current study, polyclonal antibodies directed against C YPIA I purified from rat and trout liver were used to monitor blduction of the C Y P1A 1 orthologue in hepatic microsomes from the fresh water species, the channel catfish (Ictalurus punctatus). Catfish from a local fish farm were induced in the laborato O. by three daily injections of 5Omg/kg of the PCB mixture Aroclor 1254 and compared with fish taken from a site in central Arkansas--the Bayou Meto, known to be polluted with dioxin. Hepatic microsomal activities towards ethoxyresorufin (EROD) and pentoxyresorufin (PROD) were measured and Western blot analysis carried out with the two antibodies. ERO D was elevated in both the Aroclor-treated fish and in the Bayou Meto fish compared with untreated fish farm controls; smaller but significant increases were observed in PROD. Spearman's rank correlations of 0"74 and 0"89 were observed between EROD and immunoquantified cross-reactivity towards the rat CYP1AI and trout CYPIAI antibodies,
The cytochrome P-450 system in fish, aquatic toxicology and environmental monitoring
Aquatic Toxicology, 1992
Aquatic toxicology has been defined as the qualitative and quantitative study of adverse or toxic effects of chemicals and other anthropogenic materials on aquatic organisms. The subject also includes the study of transport, distribution, transformation and ultimate fate of chemicals in the aquatic environment. Within this multidisciplinary field of science, studies of the biochemistry and function of biotransformation enzymes in aquatic organisms hold a central role. Metabolism or biotransformation through the phase 1 (cytochrome P-450 monooxygenase enzymes) and phase 11 (conjugating enzymes) pathway is a requisite for detoxification and excretion of lipophilic chemicals. In addition, such a transformation is also responsible for the activation of foreign chemicals to the intermediates that ultimately result in toxicity, carcinogenicity, and other adverse effects. The dual role of many of these enzyme systems, being involved in both xenobiotic and endogenous metabolism, furthermore makes interactions between foreign chemicals and physiological processes possible. Lastly, the response of some of these enzyme systems, in particular the cytochrome P-450 I Al subfamily, to organic xenobiotics such as oil hydrocarbons, PCBs, dioxins and dibenzofurens. makes analysis of enzyme levels by catalytic or immunochemical methods a potent way to monitor pollution effects at the molecular level. Several of these aspects will be discussed with special reference to fish.
Comparative biochemistry and physiology. Toxicology & pharmacology : CBP, 2017
The goal of this study was to determinate toxicity mechanism of biopesticide with antioxidant enzymes parameters such as superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) and malondialdehyde (MDA) levels, oxidative DNA damage (8-hydroxy-2-deoxyguanosine (8-OHdG)), transcriptional changes of heat shock protein 70 (HSP70), and cytochromes P4501A (CYP1A), sod, cat, and gpx in liver and gill tissues of Oncorhynchus mykiss. For this aim, plant-based (natural pesticides, azadirachtin (AZA)) and synthetic pesticides (deltamethrin (DLM)) were exposed on the fish at different concentrations (0.0005 and 0.00025ppm of DLM; 0.24 and 0.12ppm of AZA) for 21 days. According to the results of the study, the activity of SOD, CAT and GPx decreased, but malondialdehyde (MDA) level and activity of 8-OHdG increased in the gill and liver of rainbow trout (p<0.05). Additionally sod, cat and gpx were down regulated; HSP70 and CYP1A were up regulated for transcriptional observat...
Turkish Journal of Agriculture: Food Science and Technology, 2019
The aim of this study was to evaluate the activities of the biotransformation enzymes (Glutatyon S-Transferase (GST) and cytochrome P450 (CYP1A1)) and metabolic enzyme (Lactate dehydrogenase-LDH) in the liver of rainbow trout (Oncorhynchus mykiss) after 24 and 48 hour exposure to 0.5, 1.0 and, 2.0 mg l-1 of Congo red (CR). Enzyme activities were determined by using commercial kits with ELISA method. Congo red altered the activities of GST, CYP1A, and LDH enzymes in liver tissue of O. mykiss in a dose-dependent manner. The statistical differences in GST activities among the groups for 24 and 48 h were significant, but, LDH activities were significant for only 24 h. Exposure duration of CR didn’t affect the biochemical response of rainbow trout. Thus, CR exposure changed the biotransformation and metabolic enzymes, and the changes of these enzymes activities may be used as a potential bioindicator of the CR exposure.