139 resultados para Argyrosomus Japonicus, Microsatellites, Primer, Sciaenidae Family


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A novel solid solution Ce6MoO15 was achieved. Their structure and oxide ionic conductivity were studied. Based on Ce6MoO15, rare earth element substitution on cerium site shows that all resulting oxides enhance the conductivity further, and have high oxide-ion conductivity, which may be a kind of promising material for SOFCs.

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A total of 45 microsatellite loci from yellow perch, Perca flavescens, were isolated and characterized. Among the 45 microsatellite loci, 32 had more than two alleles. A wild population of P. flavescens (n = 48) was used to examine the allele range of the microsatellite loci. Mendelian inheritance of alleles was confirmed by examining the amplified products in pair-mated families. The number of alleles for the 32 polymorphic loci varied from two to 16, and observed heterozygosity ranged between 0.024 (YP79) and 0.979 (YP60). Cross-species polymorphic amplification in four other Percidae species was successful for 22 loci.

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Complete mitochondrial genome plays an important role in the accurate revelation of phylogenetic relationships among metazoans. Here we present the complete mitochondrial genome sequence from a sea cucumber Apostichopus japonicus (Echinodermata: Holothuroidea), which is the first representative from the subclass Aspidochirotacea. The mitochondrial genome of A. japonicus is 16,096 bp in length. The heavy strand consists of 31.8% A, 20.2% C, 17.9% G, and 30.1% T bases (AT skew = 0.027: GC skew = 0.062). It contains thirteen protein-coding genes (PCGs), twenty-two transfer RNA genes, and two ribosomal RNA genes. There are a total of 3793 codons in all thirteen mitochondrial PCGs, excluding incomplete termination codons. The most frequently used amino acid is Leu (15.77%), followed by Set (9.73%), Met (8.62%), Phe (7.94%), and Ala (7.28%). Intergenetic regions in the mitochondrial genome of A. japonicus are 839 bp in total, with three relatively large regions of Unassigned Sequences (UAS) greater than 100 bp. The gene order of A. japonicus is identical to that observed in the five studied sea urchins, which confirms that the gene order shared by the two classes (Holothuroidea and Echinoidea) is a ground pattern of echinoderm mitochondrial genomes. Bayesian tree based on the cob gene supports the following relationship: (outgroup, (Crinoids, (Asteroids, Ophiuroids, (Echinoids, Holothuroids)))). (C) 2009 Elsevier B.V. All rights reserved.

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Through random sequencing, we found a total of 884000 base-pairs (bp) of random genomic sequences in the genome of Chinese shrimp (Fenneropenaeus chinensis). Using bio-soft Tandem Repeat Finder (TRF) software, 2159 tandem repeats were found, in which there were 1714 microsatellites and 445 minisatellites, accounting for 79.4% and 20.6% of repeat sequences, respectively. The cumulative length of repeat sequences was found to be 116685 bp, accounting for 13.2% of the total DNA sequence; the cumulative length of microsatellites occupied 9.78% of the total DNA sequence, and that of minisatellites occupied 3.42%. In decreasing order, the 20 most abundant repeat sequence classes were as follows: AT (557), AC (471), AG (274), AAT (92), A (56), AAG (28), ATC (27), ATAG (27), AGG (18), ACT (15), C (11), AAC (11), ACAT (11), CAGA (10), AGAA (9), AGGG (7), CAAA (7), CGCA (6), ATAA (6), AGAGAA (6). Dinucleotide repeats, not only in the aspect of the number, but also in cumulative length, were the preponderant repeat type. There were few classes and low copy numbers of repeat units of the pentanucleotide repeat type, which included only three classes: AGAGA, GAGGC and AAAGA. The classes and copy numbers of heptanucleotide, eleven-nucleotide and thirteen-nucleotide primer-number-composed repeats were distinctly less than that of repeat types beside them.

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Growth hormone (GH), prolactin (PRL) and somatolactin (SL) were purified simultaneously under alkaline condition (pH 9.0) from pituitary glands of sea perch (Lateolabrax japonicas) by a two-step procedure involving gel filtration on Sephadex G-100 and reverse-phase high-performance liquid chromatography (rpHPLC). At each step of purification, fractions were monitored by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and by immunoblotting with chum salmon GH. PRL and SL antisera. The yields of sea perch GH, PRL and SL were 4.2, 1.0 and 0.28 mg/g wet tissue, respectively. The molecular weights of 19,200 and 20,370 Da were estimated by SDS-PAGE for sea perch GH and PRL, respectively. Two forms of sea perch SL were found: one (28,400 Da) is probably glycosylated, while the other one (23,200 Da) is believed to be deglycosylated. GH bioactivity was examined by an in vivo assay. Intraperitoneal injection of sea perch GH at a dose of 0.01 and 0.1 mug/g body weight at 7-day intervals resulted in a significant increase in body weight and length of juvenile rainbow trout. The complete sea-perch GH amino acid sequence of 187 residues was determined by sequencing fragments cleaved by chemicals and enzymes. Alignment of sea-perch GH with those of other fish GHs revealed that sea-perch GH is most similar to advanced marine fish, such as tuna, gilthead sea bream, yellowfin porgy, red sea bream, bonito and yellow tail with 98.4, 96.2%, 95.7%, 95.2%, 94.1% and 91% sequence identity, respectively. Sea-perch GH has low identity to Atlantic cod (76.5%), hardtail (73.3%), flounder (68.4%), chum salmon (66.3%), carp (54%) and blue shark (38%). Partial amino-acid sequences of 127 of sea-perch PRL and the N-terminal of 16 amino-acid sequence of sea-perch SL have been determined. The data show that sea-perch PRL has a slightly higher sequence identity with tilapia PRL( 73.2%) than with chum salmon PRL(70%) in this 127 amino-acid sequence. (C) 2001 Elsevier Science B.V. All rights reserved.

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Three cDNA sequences encoding four SNARE (N-ethylmaleimide-sensitive fusion protein attachment protein receptors) motifs were cloned from sea perch, and the deduced peptide sequences were analyzed for structural prediction by using 14 different web servers and softwares. The "ionic layer" structure, the three dimensional extension and conformational characters of the SNARE 7S core complex by using bioinformatics approaches were compared respectively with those from mammalian X-ray crystallographic investigations. The result suggested that the formation and stabilization of fish SNARE core complex might be driven by hydrophobic association, hydrogen bond among R group of core amino acids and electrostatic attraction at molecular level. This revealed that the SNARE proteins interaction of the fish may share the same molecular mechanism with that of mammal, indicating the universality and solidity of SNARE core complex theory. This work is also an attempt to get the protein 3D structural information which appears to be similar to that obtained through X-ray crystallography, only by using computerized approaches. (C) 2007 Elsevier Ltd. All rights reserved.

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Cyanobacteria are the oldest life form making important contributions to global CO2 fixation on the Earth. Phycobilisomes (PBSs) are the major light harvesting systems of most cyanobacteria species. Recent availability of the whole genome database of cyanobacteria provides us a global and further view on the complex structural PBSs. A PBSs linker family is crucial in structure and function of major light-harvesting PBSs complexes. Linker polypeptides are considered to have the same ancestor with other phycobiliproteins (PBPs), and might have been diverged and evolved under particularly selective forces together. In this paper, a total of 192 putative linkers including 167 putative PBSs-associated linker genes and 25 Ferredoxin-NADP oxidoreductase (FNR) genes were detected through whole genome analysis of all 25 cyanobacterial genomes (20 finished and 5 in draft state). We compared the PBSs linker family of cyanobacteria in terms of gene structure, chromosome location, conservation domain, and polymorphic variants, and discussed the features and functions of the PBSs linker family. Most of PBSs-associated linkers in PBSs linker family are assembled into gene clusters with PBPs. A phylogenetic analysis based on protein data demonstrates a possibility of six classes of the linker family in cyanobacteria. Emergence, divergence, and disappearance of PBSs linkers among cyanobacterial species were due to speciation, gene duplication, gene transfer, or gene loss, and acclimation to various environmental selective pressures especially light.

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To develop genetic and physical maps for shrimp, accurate information on the actual number of chromosomes and a large number of genetic markers is needed. Previous reports have shown two different chromosome numbers for the Pacific whiteleg shrimp, Penaeus vannamei, the most important penaeid shrimp species cultured in the Western hemisphere. Preliminary results obtained by direct sequencing of clones from a Sau3A-digested genomic library of P. vannamei ovary identified a large number of (TAACC/GGTTA)-containing SSRs. The objectives of this study were to (1) examine the frequency of (TAACC)(n) repeats in 662 P. vannamei genomic clones that were directly sequenced, and perform homology searches of these clones, (2) confirm the number of chromosomes in testis of P. vannamei, and (3) localize the TAACC repeats in P. vannamei chromosome spreads using fluorescence in situ hybridization (FISH). Results for objective I showed that 395 out of the 662 clones sequenced contained single or multiple SSRs with three or more repeat motifs, 199 of which contained variable tandem repeats of the pentanucleotide (TAACC/GGTTA),, with 3 to 14 copies per sequence. The frequency of (TAACC)n repeats in P. vannamei is 4.68 kb for SSRs with five or more repeat motifs. Sequence comparisons using the BLASTN nonredundant and expressed sequence tag (EST) databases indicated that most of the TAACC-containing clones were similar to either the core pentanucleotide repeat in PVPENTREP locus (GenBank accession no. X82619) or portions of 28S rRNA. Transposable elements (transposase for Tn1000 and reverse transcriptase family members), hypothetical or unnamed protein products, and genes of known function such as 18S and 28S rRNAs, heat shock protein 70, and thrombospondin were identified in non-TAACC-containing clones. For objective 2, the meiotic chromosome number of P. vannamei was confirmed as N = 44. For objective 3, four FISH probes (P1 to P4) containing different numbers of TAACC repeats produced positive signals on telomeres of P. vannamei chromosomes. A few chromosomes had positive signals interstitially. Probe signal strength and chromosome coverage differed in the general order of P1 > P2 > P3 > P4, which correlated with the length of TAACC repeats within the probes: 83, 66, 35, and 30 bp, respectively, suggesting that the TAACC repeats, and not the flanking sequences, produced the TAACC signals at chromosome ends and TAACC is likely the telomere sequence for P. vannamei.

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Given the commercial and ecological importance of the Asian paddle crab, Charybdis japonica, there is a clearly need for genetic and molecular research on this species. Here, we present the complete mitochondrial genome sequence of C. japonica, determined by the long-polymerase chain reaction and primer walking sequencing method. The entire genome is 15,738 bp in length, encoding a standard set of 13 protein-coding genes, two ribosomal RNA genes, and 22 transfer RNA genes, plus the putative control region, which is typical for metazoans. The total A+T content of the genome is 69.2%, lower than the other brachyuran crabs except for Callinectes sapidus. The gene order is identical to the published marine brachyurans and differs from the ancestral pancrustacean order by only the position of the tRNA (His) gene. Phylogenetic analyses using the concatenated nucleotide and amino acid sequences of 13 protein-coding genes strongly support the monophyly of Dendrobranchiata and Pleocyemata, which is consistent with the previous taxonomic classification. However, the systematic status of Charybdis within subfamily Thalamitinae of family Portunidae is not supported. C. japonica, as the first species of Charybdis with complete mitochondrial genome available, will provide important information on both genomics and molecular ecology of the group.

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Fully grown oocytes of Apostichopus japonicus have a cytoplasmic protuberance where the oocyte attaches to the follicle. The protuberance and the oolamina located on the opposite side of the oocyte indicate the animal-vegetal axis. Two pre-meiotic centrosomes are anchored to the protuberance by microtubules between centrosomes and protuberance. After meiosis reinitiation induced by DTT solution, the germinal vesicle (GV) migrates towards the protuberance. The GV breaks down after it migrates to the oocyte membrane on the protuberance side. The protuberance then contracts back into the oocyte and the first polar body extrudes from the site of the former protuberance. The second polar body forms beneath the first. Thus the oocyte protuberance indicates the presumptive animal pole well before maturation of the oocyte.

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With a "two-way pseudo-testcross" mapping strategy, we applied the amplified fragment length polymorphism (AFLP) markers to construct two moderate density genetic linkage maps for Laminaria. The linkage maps were generated from the 60 progenies of the F, cross family (Laminaria longissima Aresch. x L. japonica Miyabe) with twenty pairs of primer combinations. Of the 333 polymorphic loci scored in 60 progenies, 173 segregated in a 1:1 ratio, corresponding to DNA polymorphisms heterozygous in a single parent, and the other 58 loci existing in both parents followed a 3:1 Mendelian segregation ratio. Among the loci with 1:1 segregating ratios, 79 loci were ordered in 14 linkage groups (648.6 cM) of the paternal map, and 72 loci were ordered in 14 linkage groups (601.9 cM) of the maternal map. The average density of loci was approximately 1 per 8 cM. To investigate the homologies between two parental maps, we used 58 loci segregated 3:1 for further analysis, and deduced one homologous linkage group. The linkage data developed in these maps will be useful for detecting loci-controlling commercially important traits for Laminaria.

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An orange-pigmented, Gram-negative, nonmotile, strictly aerobic and oxidase- and catalase-positive bacterium (SM-A87(T)) was isolated from the deep-sea sediment of the southern Okinawa Trough area. The main fatty acids were i15 : 0, i17 : 0 3OH, i15 : 1 G, i17 : 1 omega 9c, 15 : 0, i15 : 0 3OH and summed feature 3 (comprising i-15 : 0 2OH and/or 16 : 1 omega 7c). MK-6 was the predominant respiratory quinone. DNA G+C content was 35.8 mol%. Flexirubin-type pigments were absent. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain SM-A87(T) formed a distinct lineage within the family Flavobacteriaceae, with < 93% sequence similarity to the nearest strain of genus Salegentibacter. Moreover, strain SM-A87(T) could be distinguished from the nearest phylogenetic neighbors by a number of chemotaxonomic and phenotypic properties. On the basis of polyphasic analyses, it is proposed that strain SM-A87(T) be classified in a novel genus and a new species in the family Flavobacteriaceae, designated Wangia profunda gen. nov., sp. nov. The type strain is SM-A87(T) (CCTCC AB 206139(T)=DSM 18752).

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针对目前刺参养殖过程中亟需解决的种质、环境和产品品质之间日益突出的矛盾,本论文从刺参的自身防御和内分泌机制入手,比较系统地研究了环境胁迫与细胞免疫和应激激素之间的作用机理;并从生理和分子的角度对刺参的夏眠机理进行了初步研究,主要研究成果如下: 1. 查阅国内外大量文献资料,较为系统地综述了海参的免疫体系、生态免疫学研究进展、当前夏眠机理的研究状况和刺参夏眠的基础研究。对刺参生态免疫及其夏眠机理研究提出了新的见解和思路。 2. 研究了温度(0、8、16、24、32℃)和盐度(20、25、35)的急性胁迫(72h)作用对刺参免疫指标的影响。刺参体腔液内几种免疫酶类活性受升温胁迫影响显著;而降温胁迫的影响并不显著。高盐胁迫可使刺参体腔液细胞吞噬活性、超氧化物歧化酶和过氧化氢酶活性显著变化,而对髓过氧化物酶和溶菌酶活性都没有显著影响。从这几种免疫指标变化情况来看,温度胁迫对刺参的影响要强于盐度胁迫,且高温胁迫的影响要强于低温胁迫。 3. 研究了温度(8、16、24℃)、盐度(20、25、35)和露空等急性胁迫(24h)作用对刺参体腔液儿茶酚胺类激素含量的影响。急性温度胁迫能够显著影响刺参体腔液内肾上腺素、去甲肾上腺素和多巴胺的含量;而盐度和露空胁迫对刺参体腔液肾上腺素含量、去甲肾上腺素含量以及多巴胺含量都没有产生显著影响。 4. 完成了刺参体腔液几种酶类变化的周年测定(2006年7月到2007年6月)。一年内刺参体腔液内几种酶类变化的转折点为9月份,10月份,1月份,2月份,4月份和5月份。刺参体腔液内几种酶类活性变化与温度和盐度等环境因素并不存在简单的相关关系,因此推测这些显著变化并不是单一温度、盐度等因素作用的结果,还可能与刺参生长、繁殖及其它环境因素的作用有关。 5. 开展了夏眠期间刺参免疫指标的现场研究(2006年7月到11月)。在此期间刺参体腔液细胞总数显著下降;刺参体腔液内免疫相关酶类均在8月份、9月份达到最高值;肾上腺素和去甲肾上腺素的含量均在8月下旬和11月下旬显著升高,而多巴胺含量则没有显著变化。刺参完全进入夏眠的8、9月份,体腔液内各项免疫指标也都具有显著变化,说明夏眠对刺参机体的免疫状态产生影响。 6. 探讨了刺参夏眠前后肠道基因表达的差异(2007年7月至12月)。抑制消减杂交实验结果表明接头连接效率大于50%,说明连接效率较高。消减效率分析结果表明,消减后进行cDNA保守序列扩增比未消减落后约10个循环才能明显看到扩增产物,说明对共同序列的消减效率比较高。利用抑制消减杂交技术成功构建了符合技术要求的夏眠刺参与未夏眠刺参肠道抑制消减杂交基因文库。

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本论文选择在我国分离得到的一株有毒赤潮甲藻-塔玛亚历山大藻(Alexandrium tamarense,ATHK株),研究了其对一种我国沿海常见和典型养殖鱼类鲈鱼(Lateolabrax japonicus)的危害机制。首先研究了塔玛亚历山大藻(ATHK)对鲈鱼鳃结构的影响及其溶血毒性; 然后采用腹腔注射的方法,研究了高剂量塔玛亚历山大藻毒素(ATHK毒素:约为1.6×105 细胞,相应PSP为0.886µg STX Equal,相当于每克湿重的鲈鱼PSP注射量为0.0118µg STX Equal)和低剂量塔玛亚历山大藻毒素(ATHK毒素: 约为0.16×105 细胞,相应PSP为0.0886µg STX Equal,相当于每克湿重的鲈鱼PSP注射量为0.00118µg STX Equal)在鲈鱼体内代谢过程中对鲈鱼肝脏、肾脏和鳃组织的超微结构、Na+K+-ATPase活性、肝脏功能和肾脏功能的影响、以及对抗氧化系统酶活性和异生物质代谢酶的影响,以期从不同方面了解塔玛亚历山大藻及其所产水溶性毒素(ATHK毒素)对鲈鱼的毒害效应及机制,为有毒赤潮的有效管理提供一定的科学依据。 塔玛亚历山大藻(ATHK)对鲈鱼鳃组织影响的实验结果表明,该藻使鲈鱼鳃组织出现水肿现象,细胞间隙变大;粘液细胞颗粒不规则,颜色加深,颗粒发生凝集,有板结状;氯细胞线粒体内部基质凝集。不产PSP的一种亚历山大藻(AT-6)也使鲈鱼鳃出现水肿,且使细胞出现一定的固缩现象。显微镜观察发现鲈鱼鳃丝间存在有这两种亚历山大藻细胞。由此推测塔玛亚历山大藻ATHK和AT-6的表面结构可能具有能导致鲈鱼鳃组织水肿的机械作用。对人血细胞溶血实验结果表明塔玛亚历山大藻(ATHK)具有较强的溶血毒性,大小与藻的生长阶段和细胞密度都有一定的关系:指数期的溶血毒性最大,随细胞数目的增多,活性逐渐加大;藻细胞、细胞碎片、细胞内容物都有一定的溶血毒性,其中细胞碎片的活性最大。通过11种(株)产PSP的亚历山大藻、不产PSP的亚历山大藻以及标准PSP的实验结果表明这种溶血毒性是由藻细胞的其它非PSP物质造成的,且这种溶血毒性在产PSP的亚历山大藻中具有一定的普遍性。 塔玛亚历山大藻(ATHK)对鲈鱼组织超微结构实验结果表明:ATHK毒素(约为1.6×105 细胞,相应PSP为0.886µg STX Equal,相当于每克湿重的鲈鱼PSP注射量为0.0118µg STX Equal)能导致鲈鱼组织细胞超微结构发生剧烈的变化,主要表现在:肝细胞细胞膜有肿胀现象,部分膜边缘溶解;细胞质糖原颗粒化;核糖体脱落,仅见滑面内质网;细胞质和线粒体内都出现空泡,且线粒体的嵴状结构也发生变化;核膜溶解比较严重,核质外溢,且异染色质边际化。前肾细胞超微结构的变化主要是淋巴细胞核质出现空泡,核膜有溶解迹象;Ⅰ型粒细胞颗粒膨大,伪足增多且变长;Ⅱ型颗粒细胞颗粒增多,内部出现空腔,细胞膜和核膜溶解,胞质、细胞器和核质外溢。鳃组织中氯细胞的核膜局部溶解,核仁弥散,线粒体膜溶解,微细小管膨大;粘液颗粒膜溶解,内部结构遭受破坏;扁平细胞核膜及线粒体膜几乎全部溶解。因此,我们的结果表明,ATHK毒素能作用于鲈鱼细胞的内膜和外膜系统,使膜发生溶解、脱落等变化;比较注射同样剂量大小ATHK毒素120h和240h时鲈鱼组织超微结构发现,细胞超微结构在一定程度上能够恢复。 ATHK毒素对Na+K+-ATPase活性、肝脏功能以及肾脏功能的影响结果表明,0.16×105―1.6×105细胞范围内的ATHK毒素可以显著影响肝脏和鳃组织中的Na+K+-ATPase,使这两种组织中的Na+K+-ATPase活性出现不同程度的下降; 而且还能够显著抑制肝脏中谷丙转氨酶的活性,最大抑制率为95%。但此范围内的ATHK毒素不能显著影响肾脏中的Na+K+-ATPase活性以及尿素氮含量。因此,ATHK毒素对Na+K+-ATPase活性的抑制则会导致细胞能量的缺失,使细胞进一步发生其它变化,而ATHK毒素对肝脏功能完整性的影响则可能会抑制对蛋白质的分解代谢。 ATHK毒素对鲈鱼肝脏、肾脏和鳃组织中的、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱肝肽过氧化物酶(GSH-Px)以及谷胱肝肽转硫酶(GST)活性变化影响的结果表明:高剂量(约为1.6×105 细胞,相应PSP为0.886µg STX Equal,相当于每克湿重的鲈鱼PSP注射量为0.0118µg STX Equal)ATHK毒素能显著诱导鲈鱼肝脏和鳃组织中SOD、GSH-Px以及GST酶活性,最大变化范围为正常状态下的3-4倍,对肝脏中CAT酶活性具有一定的抑制作用,对鳃中的CAT抑制效应则不显著;但此剂量的ATHK毒素仅对肾脏鳃中的GSH-Px活性有一定的诱导作用,对SOD、CAT以及GST的活性没有显著影响。低剂量(约为0.16×105 细胞,相应PSP为0.0886µg STX Equal,相当于每克湿重的鲈鱼PSP注射量为0.00118µg STX Equal)ATHK毒素也能诱导鲈鱼肝脏和鳃组织中SOD、GSH-Px以及GST酶活性,其在第一个24h内的诱导效果与临界致死毒素剂量相似; 且连续注射低剂量ATHK毒素则对肝脏和鳃中这三种酶活性具有累加的诱导作用,使这三种酶活性的变化范围为正常的5倍;低剂量ATHK毒素对肝脏中CAT酶活性也具有抑制作用,但对鳃中CAT酶活性的抑制作用并不显著。 同样,低剂量ATHK毒素除对肾脏中GSH-Px活性具有一定诱导效应外,对SOD、CAT以及GST都没有显著影响。 SOD、GSH-Px以及GST酶活性的显著升高表明ATHK毒素在鲈鱼体内代谢过程中能诱导鲈鱼产生活性氧自由基,且GST活性的升高则说明作为细胞色素P450依赖的异生物质代谢酶,GST在ATHK毒素代谢过程中可能可以加速ATHK毒素的代谢。推测鲈鱼可以通过这三种酶降低ATHK毒素以及次生毒物活性氧自由基对鲈鱼的危害。