Seasonal variation of hepatic mixed function oxidases in winter flounder (Pseudopleuronectes americanus)

Indices of hepatic mixed function oxidase (MFO) activity in winter flounder (Pseudopleuronectes americanus) were measured at approximately monthly intervals from November 1983 to October 1985. Benzo[a]pyrene hydroxylase and ethoxyresorufin O-deethylase activities and cytochromes P450 and b₅ were generally, but not significantly, higher in males than in females. The MFO activity varied seasonally, reaching a maximum during or shortly after spawning. Variation in MFO enzyme activity between sexes was never greater than 2-fold at any time, and within sex, no greater than 6-fold during the seasonal cycle. This variability is less than that caused by exposure to environmentally realistic levels of some pollutants; measurements of MFO activity in this species might therefore be used to indicate environmental contamination.     

Effects of the inhibitor ellipticine on cytochrome P450-reductase and cytochrome P450 (1A) function in hepatic microsomes of flounder (Platichthys flesus)

The effects of the mammalian inhibitor ellipticine (5,11-dimethyl-[6H]-pyrido[4,3b] carbazole) were examined in a mechanistic study of the cytochrome P450 monooxygenase system of control and β-naphthoflavone (βNF)-induced hepatic microsomes of Platichthys flesus. Ellipticine was indicated to bind to the haem moiety of cytochrome P450s (gave type II binding spectra) and to inhibit the transfer of electrons from both the hydrophobic binding site of cytochrome P450 reductase (P450R) to P450 (inhibited P450R reductase activity) and the hydrophilic binding site of P450R to soluble electron acceptors (inhibited NAD(P)H-cytochrome c reductase activity). No effect was seen on cytochrome b₅ reductase activity. Ellipticine inhibition indicated the involvement of (i) P450R (possibly also P450s) in NADPH- but not NADH- dependent hydroxyl radical production, and (ii) electron transfer and P450/P450R interaction in NADPH-dependent cytochrome P450 1A-catalysed monooxygenation (7-ethoxyresorufin O-deethylase activity and benzo(a)pyrene (BaP) metabolism). Differential effects of ellipticine on cumene hydroperoxide (CHP)-dependent BaP metabolism (P450 peroxidase activity) with CHP concentration indicated the existence of at least two forms of P450 with different substrate affinities for CHP, and different mechanisms of formation for protein adducts and free metabolites. Overall, the studies indicate the primary site of action of ellipticine in P. flesus is binding between Fe³⁺-P450 and P450R.     

柴油分散液对马粪海胆细胞色素P450 活性的影响

试验研究了不同质量浓度下(5、20、50 mg/L)三组柴油分散液,对马粪海胆的肠体、性腺、体液三个部位的抗氧化还原系统细胞色素P450活性变化的影响。结果表明,随着污染暴露时间的增加,三个部位的细胞色素P450活性均呈现出先被诱导、后被抑制的规律,并且油浓度越高,出现诱导和抑制效应的时间越早,活性变化的幅度也越大。海胆P450活性变化在一定程度上能够反映油污染的强度及其对海洋生物的毒性效应,可作为海洋石油烃污染监测的毒理学指标。     

Effects of three fungicides alone and in combination on glutathione S-transferase activity (GST) and cytochrome P-450 (CYP 1A1) in the liver and gill of brown trout (Salmo trutta)

In order to evaluate the gill glutathione S-transferase (GST) activity as a biomarker of effect of fungicide exposure in juvenile brown trout (Salmo trutta), the fungicides propiconazole {(R,S)-1-[2-(2,4-diclophenyl)-4-propyl-1,3-dioolan-2-ylmetyl]-1H-1,2,4-triazole} and fenpropimorph {(±)-cis-4-[3-(4-tert-butylphenyl)-2-metyl propyl]-2,6 dimetylmorfolinc} were administrated in the water separately and together in a static system (80 μg/l for each pesticide) for 5 days. The combined fungicides gave a significant decrease in gill GST activity towards 1-chloro-2,4-dinitrobenzene (CDNB), whilst hepatic GST-activity was not significantly changed. Furthermore, continuous exposure to 540 ug/l thiabendazole{ 2-(thiazol-4′-yl)benzimidazole} in a flow-through system for 4 days significantly increased the gill glutathione S-transferase (GST) activity towards CDNB, whilst hepatic GST and cytochrome P450 (CYP 1A) activities were not increased by the treatment.     

Effects of three fungicides alone and in combination on glutathione S-transferase activity (GST) and cytochrome P-450 (CYP 1A1) in the liver and gill of brown trout (Salmo trutta)

In order to evaluate the gill glutathione S-transferase (GST) activity as a biomarker of effect of fungicide exposure in juvenile brown trout (Salmo trutta), the fungicides propiconazole [(R,S)-1-[2-(2,4-diclophenyl)-4-propyl-1,3-dioolan-2-ylmetyl]-1H-1,2,4-triazole] and fenpropimorph [(+/-)-cis-4-[3-(4-tert-butylphenyl)-2-metyl propyl]-2,6 dimetylmorfolinc] were administrated in the water separately and together in a static system (80 microg/l for each pesticide) for 5 days. The combined fungicides gave a significant decrease in gill GST activity towards 1-chloro-2,4-dinitrobenzene (CDNB), whilst hepatic GST-activity was not significantly changed. Furthermore, continuous exposure to 540 ug/l thiabendazole[2-(thiazol-4'-yl)benzimidazole] in a flow-through system for 4 days significantly increased the gill glutathione S-transferase (GST) activity towards CDNB, whilst hepatic GST and cytochrome P450 (CYP 1A) activities were not increased by the treatment.     

Migratory behaviour of spawning rainbow trout (Oncorhynchus mykiss) in the Tongariro River,New Zealand,after habitat alteration

The movements of 92 adult rainbow trout (Oncorhynchus mykiss) from Lake Taupo into the Tongariro River, New Zealand, were monitored using radio‐telemetry every 3 days during the main spawning period between May and November 2003. This study repeated one previously conducted in 1995. Differences in spawning site preference and resting locations between 1995 and 2003 probably reflect differences in the nature of the river as a result of major natural floods that occurred in 1998. Average migration times were slower than in 1995 and the correlation between flow and mean daily movement was less distinct owing to the dry and settled weather during July and August, although fish did respond to freshets when they occurred. Peak movement occurred between 1600 and 2000 h NZ Standard Time with no movement occurring between midnight and 0400. Fish adjusted their movement in response to changing photoperiod. Nineteen fish were observed above the mouths of natal tributary streams for several weeks in the main river stem before entering these tributaries.     

小黄鱼Larimichthys polyactis体长-体重关系幂指数与产卵群体空间分布相关性研究

鱼类产卵群体的相关研究一直是渔业领域关注的热点,对鱼类的产卵期、产卵场的研究是鱼类生态习性研究方向的重要组成部分。本文在前期研究证实鱼类体长-体重关系幂指数可以指示鱼类产卵期的基础上,利用2004年4个季节的东海大面积调查数据,尝试利用同一时间点上的各个调查点幂指数信息来分析鱼类产卵场分布特征,结果发现,成熟产卵个体仅在春季出现,而未成熟产卵个体4季均有分布,以此推测小黄鱼存在"跳过产卵"现象。另外发现,单靠幂指数大小,不能区分各个产卵成熟阶段,也不能区分产卵个体与非产卵个体;而怀卵个体出现站点的幂指数平均值偏低于匀速生长,空间上也具有类似特征,即幂指数相对低的地方一般对应怀卵个体相对集中的水域。本文依据幂指数分布,推断小黄鱼成熟产卵群体有3块相对集中水域,即舟山渔场近海、济州岛西南侧和江外与舟外渔场临近水域。     

Growth hormone effect on xenobiotic-metabolizing activities of rainbow trout

In recent years several studies reported the regulation by growth hormone (GH) of the expression of a variety of P450 forms in mammals. However the effect of GH on the xenobiotic-metabolizing enzymes of fish are still unknown. The aim of this work was to investigate the effects of ovine GH—a growth hormone known to be efficient in trout—on the cytochrome P450 level and on aryl hydrocarbon hydroxylase, aminopyrine-N-demethylase, glucuronyl transferase and glutathione transferase activities in trout. The GH-implanted trout (n = 50) each received a single cholesterol pellet containing ovine GH and were compared to control animals (n = 50) receiving a single cholesterol pellet without GH. After 15 days fish were killed and the liver and gills were excised for the measurement of xenobiotic-metabolizing enzyme activities. GH treatment significantly decreased the level of hepatic cytochrome P450 and the activities of cytochrome P450 dependent monooxygenases. In contrast, no significant effect of the treatment was observed on the glutathione transferase and UDP-glucuronyl transferase activities. Moreover, GH treatment had no effect on branchial phase I and phase II enzyme activities. This study provides evidence that GH level significantly affects the expression of several members of the hepatic cytochrome P450 family in trout.     

Cytochrome P-450 isozymes in salmonids determined with antibodies to purified forms of P-450 from rainbow trout

Purification of cytochromes P-450 from liver microsomes of β-naphthoflavone (BNF)-fed rainbow trout yielded three apparently homogeneous forms. The major form (LM_4b)^* appears to be a P-448 type of cytochrome. A minor form (LM_4a), having properties very similar to LM_4b, was also obtained. In addition, a P-450 form (LM₂) was isolated, with properties quite different from LM_4a or LM_4b, including a high rate of aflatoxin B₁ (AFB₁) metabolism (Williams & Buhler, 1983c). Antibodies to all three forms were obtained from rabbits. The IgGs prepared against LM_4a and LM_4b both cross-reacted (forming lines of identity) equally well with both antigens on Ouchterlony plates. Rat P-448 cross-reacted (without lines of identity) with both LM_4a- and LM_4b-IgG. LM_4b-IgG was much more effective than LM_4a-IgG for inhibition of LM_4a or LM_4b reconstituted benzo[a]pyrene (BP) hydroxylase, suggesting that these two antibodies recognize different antigenic sites. The LM₂-IgG did not cross-react with any of the other rat or trout cytochromes P-450 examined. Levels of LM₂ and LM_4b in microsomes from untreated, polychlorinated biphenyl (PCB), phenobarbital (PB) or BNF-treated trout were estimated with an immunological technique involving electrophoresis on SDS-PAGE, followed by transfer to nitrocellulose and staining with either LM ₂ - or LM_4b -IgG. The ratio of in microsomes from PCB- or BNF-treated rainbow trout was much higher than 1, whereas the reverse was true with microsomes from untreated rainbow trout. These results are consistent with previous observations (Vodicnik et al., 1982) that pretreatment with BNF induced the synthesis of a P-448 type cyytochrome, presumably responsible for the great increase in the metabolism and activation of BP seen in these fish. Conversely, pretreatment with PB did not affect the levels of either LM₂ or LM_4b. This specific immunological technique should make it possible to assay the levels of these P-450 and P-448 isozymes in various strains of rainbow trout and other species of fish. In addition, the effect of age, sex, diet and exposure to P-450 and P-448 inducers could be examined and, perhaps, utilized to predict the relative risk of certain populations to pollutants activated by these different isozymes.     

气候变化对鲑鱼丰富度和繁殖影响的研究进展

鲑鱼是人类重要的食品来源之一,具有巨大的经济价值。同时,鲑鱼资源通过输送营养物质对维持溪流和陆地生态系统的结构、功能和过程等有重要作用。而鲑鱼是对温度极为敏感的冷水性鱼类,对气候变化的反应尤为明显。作者通过文献综述法梳理相关研究,从气候变化引起的温度变化、淡水和海洋环境的变化以及鱼病传播4个方面探讨对鲑鱼丰富度和繁殖的影响,并提出研究展望。研究显示:温度变化对鲑鱼的产卵和迁徙有较大影响;溪流和河流流量的改变会影响鲑鱼的迁徙和幼年鲑鱼的存活数量;温度和二氧化碳浓度的变化使海洋食物供应减少,鲑鱼丰富度降低;水温的升高使病原体增加,鲑鱼死亡率增加。本研究成果可为鲑鱼种群应对气候变化的研究及管理提供一定的理论支持。     

Cytochrome P4501A1 induction in Kenai River sculpin (Cottus spp.) as a monitor of freshwater pollution effects

Freshwater sculpin were taken from various locations in the Kenai River to determine their in-vivo response to pollution exposure. The specific activity of cytochrome P4501A1 was determined using fluorescence detection of the 7-ethoxyresorufin-O-dealkylase (EROD) reaction. The EROD specific activity was found to be independent of river miles but appeared to relate to specific sites of urban runoff. Glutathione-S-transferase (GST) activities were also measured but showed no significant difference between sites. Cytochrome P450 laboratory induction studies were carried out on individuals collected from a pristine site. Five groups of 10 fish were dosed from 10–150 mg β-Naphthoftavone (βNF)/kg body weight. The response was linear from 10–50 mg βNF/kg body weight. Sculpin appear to have excellent potential as a sensitive indicator of xenobiotic exposure in a freshwater benthic habitat.     

Kinetics of hepatic phase I and II biotransformation reactions in eight finfish species

Hepatic microsomes and cytosols of channel catfish (Ictalurus punctatus), rainbow trout (Oncorhynchus mykiss), Atlantic salmon (Salmo salar), red tilapia (Oreochromis sp.), largemouth bass (Micropterus salmoides), striped bass (Morone saxatilis), hybrid striped bass (M. saxatilis × M. crysops), and bluegill (Lepomis macrochuris) (n = 8) were used to study the kinetics of phase I (ECOD, EROD, PROD, BROD) and phase II (UDP-glucuronosyltransferase (UDPGT)-, sulfotransferase (ST)- and glutathione-s-transferase (GST)-mediated) reactions. The best catalytic efficiency for ECOD and GST activities was performed by channel catfish, Atlantic salmon, rainbow trout and tilapia. The highest EROD catalytic efficiency was for Atlantic salmon. None of the species had either PROD or BROD activities. Rainbow trout had very similar UDPGT catalytic efficiency to tilapia, channel catfish, Atlantic salmon, largemouth bass and bluegill. Sulfotransferase conjugation had no significant differences among the species. In summary, tilapia, channel catfish, Atlantic salmon and rainbow trout had the best biotransforming capabilities; striped bass, hybrid striped bass and bluegill were low metabolizers and largemouth bass shared some capabilities with both groups.     

Effects of β-naphthoflavone on hepatic biotransformation and glutathione biosynthesis in largemouth bass (Micropterus salmoides)

We are investigating the effects of in vivo exposure of prototypical enzyme inducing agents on hepatic biotransformation enzyme expression in largemouth bass (Micropterus salmoides), a predatory game fish found throughout the United States and Canada. The current study targeted those genes involved in biotransformation and oxidative stress that may be regulated by Ah-receptor-dependent pathways. Exposure of bass to β-naphthoflavone (β-NF, 66 mg/kg, i.p.) elicited a 7–9-fold increase in hepatic microsomal cytochrome P4501A-dependent ethoxyresorufin O-deethylase (EROD) activities, but did not affect cytosolic GST catalytic activities toward 1-chloro-2,4-dinitrobenzene (CDNB) or 5-androstene-3,17-dione (ADI). Glutathione S-transferase A (GST-A) mRNA expression exhibited a transient, but non-significant increase following exposure to β-NF, and generally tracked the minimal changes observed in GST–CDNB activities. Expression of the mRNA encoding glutamate-cysteine ligase catalytic subunit (GCLC), the rate-limiting enzyme in glutathione (GSH) biosynthesis, was increased 1.7-fold by β-NF. Changes in GCLC mRNA expression were paralleled by increases in intracellular GSH. In summary, largemouth bass hepatic CYP1A-dependent and GSH biosynthetic pathways, and to a lesser extent GST, are responsive to exposure to β-NF.     

Indications of P450 monooxygenase activities in beluga (Delphinapterus leucas) and narwhal (Monodon monoceros) from patterns of PCB, PCDD and PCDF accumulation

Selective accumulation of PCDDs, PCDFs and PCB congeners was investigated in beluga (Delphinapterus leucas), narwhal (Monodon monoceros) and several species at the same trophic level in two areas of Canada. There was a much higher relative abundance of meta-para-unsubstituted PCBs in the cetaceans compared with other species. This suggests that activity of cytochrome P450 monooxygenase enzymes causing CYP2B-type metabolism was relatively low in beluga and narwhal. Lower relative levels of 2,3,7,8-TCDD occurred in beluga from both areas, and selective reduction of toxic non-ortho PCBs occurred in the highly PCB-contaminated St Lawrence beluga. These compounds are potent inducers of cytochrome P450 CYP 1A-type enzymes, suggesting that such an enzyme with the capability of metabolizing TCDD-like substrates is present in beluga and narwhal. SIMCA principal components analysis of PCB congener patterns showed that the two cetaceans were in a statistically significant different class than the other species. Variables (congeners) with the greatest separation power between classes were mainly those defined by the selective metabolism rules.     

Time-course studies of the biotransformation enzymes in control rainbow trout when adjusting to new habitats

Hepatic monooxygenase enzyme activities and relative cytochrome P4501A protein content were measured to evaluate the time-course alterations in rainbow trout after change in living habitat. Fish were transferred from one fish farm to the tanks of another hatchery and/or into cages kept in a lake. In the new habitats, cytochrome P450-dependent enzyme activities in rainbow trout decreased, and were at their lowest levels after two or three weeks in the summer. Later the activities partly reversed. The immunodetection of cytochrome P4501A protein expressed a similar trend as for catalytic monooxygenase activities.     

Paralytic shellfish poisoning toxins induce xenobiotic metabolising enzymes in Atlantic salmon (Salmo salar).

Paralytic shellfish poisoning (PSP) toxins have been implicated as the causative agent of a number of fish kills. Exposure experiments indicate that fish are susceptible to PSPs by intraperitoneal (i.p.) and oral administration, while sampling of fish affected by toxic blooms reveals that these toxins can be accumulated. In spite of the potential impact to marine fisheries, little research has been conducted on the potential metabolism and detoxification of PSPs in marine fishes. Previous work by this group has shown that the xenobiotic metabolising enzyme (XME) cytochrome P-450 (CYP1A) is induced in Atlantic salmon (Salmo salar) following i.p. exposure to saxitoxin (STX). Salmon injected i.p. with sub-lethal doses of STX show a four- to eight-fold induction of hepatic CYP1A (as shown by ethoxyresorufin-O-deethylase activity) over controls after 96 h. Results presented here show that the phase II XME glutathione S-transferase (GST) is also induced in salmon following PSP exposure. Post smolts were exposed to three injections of PSPs (2 micrograms STXeq/kg) over 21 days. Injection of both STX and PSPs extracted from a toxic strain of dinoflagellate (Alexandrium fundyense, CCMP 1719) resulted in induction of hepatic GST, as measured by activity for 1-chloro 2,4-dinitrobenzene. Such inductions indicate a potential role for XMEs in PSP metabolism. Possible roles for other enzymes are also discussed.     

Seasonal variation of oxidative biomarkers in gills and digestive gland of green-lipped mussel Perna viridis from Arabian Sea

Investigations on seasonal variation in oxidative stress biomarkers were carried out on the natural population of green-lipped mussel Perna viridis collected from Bambolim beach area of Goa. Oxidative stress indices such as lipid peroxidation (LPX), hydrogen peroxide (H₂O₂), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione transferase (GST), glutathione reductase (GR), reduced glutathione (GSH) and ascorbic acid (ASA) were measured in gills and digestive gland of P. viridis during February, May, August and November. The present study reveals two important aspects regarding the antioxidant defence status of tissues of P. viridis. Firstly, antioxidant capacity of tissues of P. viridis exhibits seasonal variation. Secondly, various components of antioxidant capacity such as oxidative stress markers, levels of antioxidant enzymes and small antioxidant molecules vary differently in tissues with respect to different seasons. Although the oxidative stress status of gills and digestive gland of P. viridis expressed in terms of LPX and H₂O₂ was the lowest in February, its level was maximal in gills and digestive gland during May and November, respectively. While activities of SOD and GPX of tissues of P. viridis were found to be low in August, activities of CAT and GR were recorded to be low in February. GST activity in gills although remained high in February, in digestive gland elevated values were recorded in August and November. A seasonal variation in the levels of small antioxidant molecules was also noticed. Among non enzymatic antioxidants ASA content of tissues was maximal in May and August in comparison to February and November, but GSH remained high in November. It therefore appears that environmental factors may play a crucial role in regulating the oxidative stress capacity of tissues of P. viridis.     

Migration dynamics of Pacific herring (Clupea pallasii) and response to spring environmental variability in the southeastern Bering Sea

In the southeastern Bering Sea, Pacific herring (Clupea pallasii) migrate from the Pribilof Islands region where they overwinter, to the Alaska coast where they spawn in spring. The migration sustains a nearshore commercial fishery that targets roe-bearing females just prior to spawning. Herring also are taken as bycatch in groundfish trawl fisheries, where time and area closures in these fisheries are triggered by herring bycatch caps. Using herring bycatch data collected since the 1970s by National Marine Fisheries Service (NMFS) observers aboard groundfish fishing vessels, a retrospective analysis was conducted to describe the seasonal migration pattern of Pacific herring in the southeastern Bering Sea and to study its spatial and temporal variability. Observed changes in herring catch per unit of effort were compared with variability in climate and oceanographic conditions. The seasonal migration is complex, but annual shifts in migration routes and a possible northward shift of the overwintering grounds was identified. Pre-spawning herring aggregated in different areas depending on whether spawning occurred early or late in spring. The thermal structure of the ocean around the ice edge appears to influence herring migration timing and route as well as spawning date. Thus, on the basis of recent changes in sea-ice extent and duration, we suggest that the herring bycatch savings area that was developed from data collected in the 1980s should be revised to reflect prevailing conditions.     

Quantification of cytochrome P4501A1 and catalytic activities in liver microsomes of isosafrol- and β-Naphthoflavone-treated rainbow trout (Oncorhynchus mykiss)

The effects of isosafrol (ISF) or β-naphthoflavone (βNF) treatments on cytochrome P450 (P450) levels in rainbow trout liver were investigated using immunochemical and catalytic techniques. The discrepancies in catalytic activities and ELISA quantification of rainbow trout P4501A1 protein levels between ISF- and βNF-treated fish indicate that important differences exist between the responses induced by βNF and ISF treatments in the rainbow trout liver.