Apoptosis-inducing protein, AIP, from parasite-infected fish induces apoptosis in mammalian cells by two different molecular mechanisms

AIP (apoptosis-inducing protein) is a protein purified and cloned from Chub mackerel infected with the larval nematode, Anisakis simplex, which induces apoptosis in various mammalian cells including human tumor cell lines. AIP has shown structural and functional homology to L-amino acid oxidase (LAO) which oxidizes several L-amino acids including L-lysine and AIP-induced apoptosis has been suggested to be mediated by H2O2 generated by LAO activity of AIP. In this study, we confirmed that recombinant AIP generated enough H2O2 in culture medium to induce rapid apoptosis in cells and this apoptosis was clearly inhibited by co-cultivation with antioxidants such as catalase and N-acetyl-cysteine. Surprisingly, however, we found that AIP still could induce H2O2-independent apoptosis more slowly than H2O2-dependent one in HL-60 cells even in the presence of antioxidants. In addition, the HL-60-derived cell line HP100-1, which is a H2O2-resistant variant, underwent apoptosis on treatment with AIP with a similar delayed time course. The latter apoptosis was completely blocked by addition of L-lysine to the culture medium, which is the best substrate of AIP as LAO, indicating that decreased concentration of L-lysine in the culture medium by AIP-treatment induced apoptosis. We also showed that the both apoptosis by AIP were associated with the release of cytochrome c from mitochondria and activation of caspase-9, and overexpressed Bcl-2 could inhibit both of the AIP-induced apoptosis. These results indicate that AIP induces apoptosis in cells by two distinct mechanisms; one rapid and mediated by H2O2, the other delayed and mediated by deprivation of L-lysine, both of which utilize caspase-9/cytochrome c system.     

Evaluation of Entomopathogenic Nematodes for the Control of Mediterranean Fruit Fly (Diptera: Tephritidae)

The virulence of various entomopathogenic nematode (EPN) strains was evaluated against the Mediterranean fruit fly, C. capitata . The selected nematodes were assessed for their infectivity for the final larval stage of the insect host and under varying environmental conditions. Among 12 EPN strains tested, Steinernema riobrave Texas ( Sr TX) and Heterorhabditis sp. IS-5 (H IS-5), showed high activity and induced >80% mortality. Six EPN strains showed limited activity (>30% mortality), and four strains had no effect (<20% mortality). Sr TX was more effective than H IS-5. Mature C. capitata larvae were most susceptible to nematode infection during the first 4h after they began to emerge from their diet to pupate. Activity of the two nematode strains at a constant inoculation rate was dependent on insect larval density. The highest activity was recorded at 1.88 larvae cm -2 and decreased at higher larval densities. EPN activity was also directly related to nematode density. Maximal activity was shown at a density of 150 infective juveniles cm -2 . A similar activity pattern was also recorded with Sr TX in four different soil types. The persistence of this EPN in the soil extended over 5 days but there was no activity after 14 days. Except for a lower activity under cool conditions (17°C), temperatures ranging between 22 and 41°C, or moisture levels in the treated soil ranging between 3 and 20%, had no significant effect on nematode activity. Our results suggest that application of Sr TX against C. capitata may have potential for controlling C. capitata .     

hDKIR, a human homologue of the Drosophila kelch protein, involved in a ring-like structure

We have previously purified and cloned an apoptosis-inducing protein (AIP) derived from fish infected with the anisakis simplex. Recently, we identified a series of AIP-responsive genes in the HL-60 cell line using a subtractive hybridization method. Here we report the molecular cloning and characterization of one of these genes, which encodes a novel human kelch protein containing 568 amino acid residues, termed hDKIR. The Drosophila Kelch protein localizes to a ring canal structure, which is required for oocyte development. When hDKIR was expressed in cultured-mammalian cells, hDKIR localized to a ring-like structure. Furthermore, when coexpressed with Mayven or Keap1, hDKIR bound to Mayven and recruited Mayven into ring-like structures perfectly. This indicates that kelch homologues can interact with each other in a specific manner and such interaction can affect the subcellular localization of kelch proteins. Finally, domain analysis revealed that both the N-terminal POZ (poxviruses and zinc fingers) and intervening region (IVR) domains of hDKIR are essential for ring-like structure activity, suggesting that the development of the ring-like structure is independent of the ability to bind actin.     

In vitro bioassay of sheep gastrointestinal mucus for nematode paralysing activity mediated by substances with some properties characteristic of SRS-A

A bioassay is described for determining the inhibition of nematode larval migration from agar by substances exerting a paralysing action.In the assay, larval migration was completely inhibited by the anthelmintic levamisole (25 μg/ml) whereas biogenic amines, and prostaglandins E₁ and E₂ at 50 μg/ml, were without effect. Mucus from the gastrointestinal tract of sheep resistant to nematode infection inhibited larval migration by up to 93% whereas mucus from heavily infected sheep or sheep reared helminth free did not significantly inhibit larval migration. Mucus from sheep resistant to Trichostrongylus colubriformis inhibited the migration of larvae of other nematode species.The larval migration inhibitory (LMI) activity of mucus from resistant sheep was associated with components having some properties of slow reacting substance of anaphylaxis (SRS-A).Faecal samples from resistant sheep possessed significantly more LMI activity than faecal samples from heavily infected sheep or sheep reared helminth free. The level of LMI activity in the faeces of sheep undergoing challenge infection may be a useful indicator of the sheep's resistance status.The presence of the larval migration inhibitory activity in sheep mucus is discussed in relation to resistance to infection.     

The two actin-interacting protein 1 genes have overlapping and essential function for embryonic development in Caenorhabditis elegans

Disassembly of actin filaments by actin-depolymerizing factor (ADF)/cofilin and actin-interacting protein 1 (AIP1) is a conserved mechanism to promote reorganization of the actin cytoskeleton. We previously reported that unc-78, an AIP1 gene in the nematode Caenorhabditis elegans, is required for organized assembly of sarcomeric actin filaments in the body wall muscle. unc-78 functions in larval and adult muscle, and an unc-78-null mutant is homozygous viable and shows only weak phenotypes in embryos. Here we report that a second AIP1 gene, aipl-1 (AIP1-like gene-1), has overlapping function with unc-78, and that depletion of the two AIP1 isoforms causes embryonic lethality. A single aipl-1-null mutation did not cause a detectable phenotype. However, depletion of both unc-78 and aipl-1 arrested development at late embryonic stages due to severe disorganization of sarcomeric actin filaments in body wall muscle. In vitro, both AIPL-1 and UNC-78 preferentially cooperated with UNC-60B, a muscle-specific ADF/cofilin isoform, in actin filament disassembly but not with UNC-60A, a nonmuscle ADF/cofilin. AIPL-1 is expressed in embryonic muscle, and forced expression of AIPL-1 in adult muscle compensated for the function of UNC-78. Thus our results suggest that enhancement of actin filament disassembly by ADF/cofilin and AIP1 proteins is critical for embryogenesis.     

A novel apoptosis-inducing protein from Helicobacter pylori

Helicobacter pylori infection induces apoptosis in gastric epithelial cells. Here, we report a novel apoptosis-inducing protein that functions as a leading factor in H. pylori-mediated apoptosis induction. We purified the protein from H. pylori by separating fractions that showed apoptosis-inducing activity. This protein induced apoptosis of AGS cells in a dose-dependent manner. The purified protein consisted of two protein fragments with molecular masses of about 40 and 22 kDa, which combined to constitute a single complex in their natural form. N-terminal sequencing indicated that both these protein fragments were encoded by the HP1118 gene. The purified protein exhibited gamma-glutamyl transpeptidase activity, the inhibition of which by 6-diazo-5-oxo-l-norleucine resulted in a complete loss of apoptosis-inducing activity. To the best of our knowledge, the apoptosis-inducing function is a newly identified physiological role for bacterial gamma-glutamyl transpeptidase. The apoptosis-inducing activity of the isogenic mutant gamma-glutamyl transpeptidase-deficient strain was significantly lower compared with that of the parent strain, demonstrating that gamma-glutamyl transpeptidase plays a significant role in H. pylori-mediated apoptosis. Our findings provide new insights into H. pylori pathogenicity and reveal a novel aspect of the bacterial gamma-glutamyl transpeptidase function.     

Modelling regional differences in 'sealworm', Pseudoterranova decipiens (Nematoda, Ascaridoidea), infections in some North Atlantic cod, Gadus morhua, stocks

The ascaridoid nematode Pseudoterranova decipiens has a complex life-cycle. Adult worms reproduce in the stomach of grey seals, Halichoerus grypus , and other pinnipeds, and the larvae are found encysted in the musculature of Atlantic cod, a common host. An increase in infection levels in Canadian, Icelandic and Norwegian cod stocks over the last decade has been attributed to recent increases in seal population numbers. A simple model is proposed for the accumulation of larval parasites in the fish through its food. The model describes how regional differences in fish growth can also explain some differences in infection levels.     

'Cebiche'--a potential source of human anisakiasis in Mexico?

Five fish species used for preparation of a popular dish (cebiche) made with raw fish flesh in Mexico were obtained from five localities of the coast of Yucatan. Lutjanus synagris, Gerres cinereus, Sphyraena barracuda, Epinephelus morio and Haemulon plumieriwere examined for the presence of larvae of anisakid nematodes, causative agents of human anisakiasis. The nematode Pseudoterranova sp. was found in E. morio and S. barracuda with a total prevalence of 83% and 6.5 +/- 6.2 worms per fish for E. morio, and a prevalence of 33% and 10.2 +/- 30.0 worms per fish for S. barracuda. Contracaecumsp. was found to infect G. cinereus with a prevalence of 57% and 7.6 +/- 11.4 worms per fish. The relatively high prevalence of Pseudoterranova sp. indicates that this parasite is a potential causal agent of anisakiasis on the coast of Yucatan. Although all larvae were found only in the mesentery of the fish host, their importance as a potential source of human infection cannot be excluded as larval migration to the muscles in dead fish is possible.     

The diet of blue whiting, hake, horse mackerel and mackerel off Portugal

This paper deals with the diets of blue whiting Micromesistius poutassou (Risso 1810), hake Merluccius merluccius (L. 1758), horse mackerel Trachurus trachurus (L. 1758), and mackerel Scomber scombrus (L. 1758) off Portugal and explores variations in fish length, water depth, latitude and season. All four species feed on fish; however, hake and mackerel are the first and second most important predators, respectively, blue whiting being the most important fish prey for both species. The diets of blue whiting and horse mackerel are composed mainly of crustaceans. Diet variations according to predator fish size are more important than either latitude or depth. In the diets of blue whiting, hake and horse mackerel, prey importance increases with predator size. For blue whiting and horse mackerel, diet variations with fish length and water depth are correlated: small fish are closely associated with coastal areas where they feed on copepods and decapod larvae. Seasonality in the diet is apparent for blue whiting, hake and mackerel. For blue whiting, the decapod Pasiphaea sivado is the most important prey in summer and autumn, being replaced by the euphausid Meganyctiphanes norvegica in winter. In the diet of hake, seasonality was characterised by the major importance of Macroramphosus scolopax in autumn, whereas the diet of mackerel consisted of zooplankton in summer, fish and decapods in autumn and decapod larvae in winter. Seasonal changes in the diet of horse mackerel correspond to a higher diversity of prey in autumn compared to other seasons (although euphausids are the main prey in all seasons). Seasonality in feeding activity is not as marked for the other species as it is for horse mackerel; the percentage of empty stomachs of horse mackerel is greatest in winter, when spawning takes place at the Portuguese coast.     

Morphological and molecular diagnosis of anisakid nematode larvae from cutlassfish (Trichiurus lepturus) off the coast of Rio de Janeiro, Brazil

Anisakid nematode larvae from Trichiurus lepturus off coast of Rio de Janeiro were studied using light, laser confocal and scanning electron microscopy, in addition to a molecular approach. Mitochondrial cytochrome c-oxidase subunit 2 (mtDNA cox-2), partial 28S (LSU) and internal transcribed spacers (ITS-1, 5.8S, ITS-2) of ribosomal DNA were amplified using the polymerase chain reaction and sequenced to evaluate the phylogenetic relationships between the nematode taxa. The morphological and genetic profiles confirmed that, of the 1,030 larvae collected from the 64 fish examined, 398 were analysed, of which 361 were Hysterothylacium sp. and 37 were Anisakis typica. Larvae of Hysterothylacium sp. were not identified to the species level due to the absence of similar sequences for adult parasites; however, the ITS sequence clustered in the phylogenetic tree with sequences of H. deardorffoverstreetorum, whereas an mtDNA cox-2 and LSU concatenated phylogenetic analysis demonstrated the presence of two clades, both of them under the same name as the larval H. deardorffoverstreetorum. Data on the occurrence of parasites during the winter and summer months were compared using the t-test. The greatest prevalence and intensity of infection were recorded for larval Hysterothylacium, with a prevalence of 51.56% and an intensity of up to 55 parasites per fish. The larval Anisakis exhibit a higher abundance and intensity of infection in the winter months, and those of Hysterothylacium during the summer. However, the t-test indicated no significant differences between the abundance and intensity of infection recorded during the months of collection for either of these larval nematodes. All sequences generated in this study were deposited in GenBank.     

Toxicity of Bacillus thuringiensis to parasitic and free-living life-stages of nematode parasites of livestock

A collection of Bacillus thuringiensis (Bt) strains (Bts) were screened for activity against the free-living larval stages of nematode parasites of livestock. Two strains were identified with significant activity in inhibiting larval development of Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia circumcincta. These strains were also toxic to the adult parasitic stages of these nematode species in vitro. Adult H. contortus and O. circumcincta showed complete cessation of movement within 2 and 4 days, respectively. Trichostrongylus colubriformis adults were less affected, however, movement was still significantly reduced compared with controls. The in vitro activity against the larval stages was of a magnitude similar to or greater than that seen with the anthelmintic drugs thiabendazole and levamisole. N-terminal amino acid sequencing indicated that the two Bts contained either Cry5A and Cry5B proteins, or a Cry13 protein, and the presence of the corresponding cry5A, cry5B and cry13 genes was confirmed by PCR and sequencing. Bacillus thuringiensis spore-crystal suspensions exposed to acidic pH conditions (pH相似文献   

Anisakis simplex: CO(2)-fixing enzymes and development throughout the in vitro cultivation from third larval stage to adult

We studied the effect of CO(2) on the in vitro cultivation of Anisakis simplex, an aquatic parasitic nematode of cetaceans (final hosts) and fish, squid, crustaceans and other invertebrates (intermediate/paratenic hosts), and, occasionally, of man (accidental host). The results showed that a high pCO(2), at a suitable temperature, is vital for the optimum development of these nematodes, at least from the third larval stage (L3) to adult. After 30 days cultivation in air, molting to L4 (fourth larval stage) was reduced to 1/3, while survival was about 1/3 of that when cultivated in air + 5% CO(2). The activity of the CO(2)-fixing enzymes, PEPCK and PEPC, was also studied. Throughout the development of the worms studied, PEPCK activity was much higher than that of PEPC (e.g., 305 vs. 6.8 nmol/min.mg protein, respectively, in L3 collected from the host fish). The activity of these enzymes in the worms cultivated in air + 5% CO(2) was highest during M3, and was also generally higher than that of those cultivated in air only, especially during molting from L3 to L4 (e.g., in recently molted L4, PEPCK activity was 3.7 times greater than that of PEPC 2.9 times greater than when cultivated in air).     

The effect of vaccinating infection during pregnancy and dietary protein supply on the peri-parturient immune response of sheep to infection with Teladorsagia circumcincta and Trichostrongylus colubriformis larvae

It is well established that dietary protein supply can influence the peri-parturient breakdown of immunity to nematode parasites but there is no information on the importance of exposure to nematode larvae during pregnancy for this response. We investigated this by exposing housed pregnant sheep, scanned as carrying two lambs, to a vaccinating infection with a trickle mixed infection of Teladorsagia circumcincta and Trichostrongylus colubriformis larvae (L3) or to no infection during weeks − 9 to − 4 relative to parturition. At the beginning of week − 3 all sheep were treated with anthelmintic to remove any vaccinating worm burden and from week − 2 to week +6 received a trickle challenge infection with the same nematodes. Within each vaccinating treatment there were two nutritional treatments (no. = 20 per subgroup) designed to provide 1.5 or 1.0 and 1.3 or 0.8 of metabolisable protein (MP) requirement during pregnancy and lactation, respectively. Five ewes were necropsied during weeks +1 and +3 to measure worm burdens and mucosal inflammatory cells and the remainder maintained until week +6. Serum levels of total, IgA and IgE antibodies against L3 antigen of each nematode were measured.Scanning errors and lamb losses resulted in some ewes carrying and/or rearing only one lamb. Numbers of lambs reared was therefore introduced as a treatment effect. Vaccinating infection delayed the peri-parturient rise in faecal egg count (FEC) by an average of 2 weeks but its effect on FEC during the first 6 weeks of lactation was smaller and less persistent than that of dietary MP supply and single- v. twin-suckling.Populations of both nematodes were lower in association with high MP supply, vaccination and single suckling. These changes were associated with increases in numbers of mucosal mast cells (MMC) as a result of both increased MP supply and vaccination. Evidence for a more rapid return of host ability to limit populations of the abdominal nematode T. circumcincta than of the intestinal nematode T. colubriformis was associated with fewer eosinophils and more globule leucocytes (GL) in abomasal than in intestinal tissue.None of the serum antibody isotypes was affected by dietary protein supply. Total and IgA antibodies were maintained by a current larval (vaccinating) intake. IgA titres, however, increased progressively during pregnancy, especially in twin-bearing ewes. IgE titres appeared to be sensitive primarily to the reproductive cycle itself, peaking around parturition.This work supports the conclusion that availability of MP supply influences the recruitment and activity of cells of the immune armoury of the gastro-intestinal tract to nematode parasites. The precise outcome may differ with site and/or nematode species.     

Chromosomal polymorphism is associated with nematode parasitism in a natural population of a tropical midge

A natural population of a tropical midge, Chironomus ramosus (Diptera: Chironomidae), was found to be polymorphic for a paracentric inversion (IV: 18C-19D). Based on the characteristic banding pattern of the fourth chromosome in the larval salivary gland polytene nuclei, individuals were classified as either structural homozygotes or heterozygotes. Isofemale lines were obtained and subsequently standard (S/S) and inversion (I/I) homozygotes were characterised by careful progeny testing in the laboratory. While exploring various biotic and abiotic factors that might be responsible for the maintenance of inversion polymorphism, we detected nematode (Family: Mermithidae) infections among the larval population. A detailed study indicated that the inversion polymorphism in the natural population of C. ramosus was apparently being maintained as a result of the selective pressure exerted by the nematode parasite. The corresponding pattern of increase and decrease in genotype frequencies and the relative fitness values indicated a selective advantage of inversion heterozygotes (S/I) over both homozygous types (S/S and I/I). Both empirical and experimental data suggest the strong heterotic nature of adaptation in this C. ramosus population towards nematode infection. This is the first report of its kind where inversion polymorphism has been shown to be associated with nematode parasitism.     

Conflict between antipredator and antiparasite behaviour in larval damselflies

 Larval damselflies resist infestation by parasitic larval mites by exhibiting behaviours such as grooming, crawling, swimming, and striking at host-seeking mites. Larval damselflies are known to increase time spent in these behaviours in the presence of mites but reduce time spent in these behaviours in the presence of fish predators. The presence of both fish and larval mites presents an obvious conflict: a larval damselfly may actively avoid parasitism by mites, thus increasing its risk of predation, or it may reduce its activity when fish are present, thus increasing its risk of parasitism. We analysed the behaviour of larval Ischnura verticalis in an experiment where we crossed presence and absence of fish with presence and absence of larval mites. Presence of mites induced a large increase in activity of larval I. verticalis but fish had no effect and there were no interpretable interactions between effects of mites and fish. Subsequent experiments indicated that larval I. verticalis in the presence of both mites and fish were more likely to be attacked and killed by fish than those exposed only to fish. The high activity level of I. verticalis larvae in the presence of both fish and mites may suggest that costs of parasitism are high, or that under field conditions it is rare for larvae to be in the immediate presence of both fish predators and potentially parasitic mites. Received: 28 March 1996 / Accepted: 6 September 1996     

Microhabitat selection by larval Chironomus tentans (Diptera: Chironomidae): effects of predators, food, cover and light

1. As part of a study designed to estimate the developmental costs of antipredator behaviour of larval chironomids, we used laboratory experiments to study effects of food and factors that could influence predation risk [presence of fish, cover from fish (simulated debris) and light level], on microhabitat selection by Chironomus tentans larvae in the third and fourth instar. 2. Larvae were more likely to build tubes where there was more food although their ability to move far to find food appeared limited. 3. Larvae did not avoid areas with fish and the presence of fish did not alter larval response to food. 4. Larvae avoided areas of cover (simulated debris) but cover did not alter larval response to food. 5. When provided with a choice between light and dark areas, larvae initially without tubes were found more often in the dark areas. Light level had no effect on location of larvae that had begun the experiment with tubes. 6. Results suggest the tubicolous life-style of larval Chironomus tentans limits their ability to select microhabitats that could alter their risk of predation.     

The intensity and duration of primary Heligmosomoides polygyrusinfection in TO mice modify acquired immunity to secondary challenge

The effect of dose and duration of immunizing infections of Heligmosomoides polygyrus on protection against homologous challenge was studied in female TO mice. Primary infections were terminated at various levels with pyrantel embonate (adult infections) or ivermectin (larval infections) and mice were then challenged with 500 infective larvae (L3). The level of protection to secondary challenge positively correlated with the intensity of the primary immunizing infection but truncation of larval infection produced significantly better protection than termination of the adult nematode infection. The duration of the primary larval infection (1-6 days) positively correlated with the level of protection to secondary challenge, antibody responses and the proportion of circulating eosinophils. Histological changes in the gastrointestinal tract, peripheral leucocytic changes and antibody responses of the mice to H. polygyrus adult somatic antigens indicate both a cellular and humoral basis of host immunity to secondary challenge. Although the TO mice are slow responders in that they harbour chronic infections, immunization by intramucosal killing of the larval stage produced strong protection against secondary challenge infection. The presence of dead immunogenic larval stages within the intestinal wall may well be an important factor, since it exposes the host to stage specific antigens at an appropriate location. The implications of the findings for the control of gastrointestinal nematode infections are also discussed.     

AIP is a mitochondrial import mediator that binds to both import receptor Tom20 and preproteins

Most mitochondrial preproteins are maintained in a loosely folded import-competent conformation by cytosolic chaperones, and are imported into mitochondria by translocator complexes containing a preprotein receptor, termed translocase of the outer membrane of mitochondria (Tom) 20. Using two-hybrid screening, we identified arylhydrocarbon receptor-interacting protein (AIP), an FK506-binding protein homologue, interacting with Tom20. The extreme COOH-terminal acidic segment of Tom20 was required for interaction with tetratricopeptide repeats of AIP. An in vitro import assay indicated that AIP prevents preornithine transcarbamylase from the loss of import competency. In cultured cells, overexpression of AIP enhanced preornithine transcarbamylase import, and depletion of AIP by RNA interference impaired the import. An in vitro binding assay revealed that AIP specifically binds to mitochondrial preproteins. Formation of a ternary complex of Tom20, AIP, and preprotein was observed. Hsc70 was also found to bind to AIP. An aggregation suppression assay indicated that AIP has a chaperone-like activity to prevent substrate proteins from aggregation. These results suggest that AIP functions as a cytosolic factor that mediates preprotein import into mitochondria.     

匙吻鲟仔稚鱼消化酶发育的研究

对出膜后0—53d匙吻鲟的酸性蛋白酶、碱性蛋白酶、α-淀粉酶、脂肪酶以及磷酸酶的活性变化进行了测定。匙吻鲟出膜后饲养于室内水泥培育池中,从第3天开始投喂枝角类,之后于第40天将试验鱼转移至池塘。试验材料为受精卵及出膜后第3、第6、第12、第20、第30、第40、第44、第47、第53天仔稚鱼样品。研究发现主要消化酶在出膜时或卵黄期即可检测出活力。碱性蛋白酶和酸性蛋白酶分别在出膜后3d(3DAH)和刚出膜时(0DAH)检测出活力。碱性蛋白酶活力在44DAH达到最大值[(1.96±0.09)U/fish],47DAH出现下降,但在53DAH开始上升,比活力在53DAH达到最大值[(8.84±0.59)U/mg protein]。酸性蛋白酶在44DAH达到最大值[(0.52±0.05)U/fish],比活力在6DAH出现第一个峰值[(2.08±0.09)U/mg protein],并在30DAH出现最小值[(0.83±0.06)U/mg protein]。试验期间碱性蛋白酶活力高于酸性蛋白酶。在12DAH—40DAH期间α-淀粉酶活力相对稳定,并在47DAH达到最大值[(0.42±0.03)U/fish],比活力在12DAH出现一个峰值[(1.18±0.12)U/mg protein],并于47DAH出现最大值[(1.94±0.16)U/mg protein]。发育早期脂肪酶活力较高,活力和比活力分别在30DAH[(0.20±0.02)U/fish]和6DAH[(2.28±0.22)U/mg protein]出现最大值。碱性磷酸酶活力变化趋势与比活力变化趋势相似,但是最大值分别出现在44DAH[(0.08±0.00)U/fish]和30DAH[(1.96±0.15)U/mg protein]。酸性磷酸酶活力在3DAH出现一个峰值[(0.01±0.00)U/fish],之后显著升高,并在44DAH达到最大值[(0.05±0.00)U/fish],其比活分别在30DAH[(1.19±0.10)U/mg protein]和44DAH[(1.10±0.08)U/mg protein]出现两个峰值。结果表明,蛋白酶、α-淀粉酶和磷酸酶随个体发育活力增加,碱性蛋白酶在个体发育早期对蛋白质的消化具有重要作用。养殖环境发生改变时,酸性蛋白酶、α-淀粉酶、碱性磷酸酶和酸性磷酸酶活力在生长减慢时增加,生长加快时降低,而脂肪酶活力则维持稳定。