斑点叉尾趋化因子SCYA126基因及其可变剪接

通过探针杂交筛选斑点叉尾(Ictalurus punctatus)BAC基因组文库,利用引物步移的方法对阳性克隆BAC047 K12进行序列测定,得到2 815 bp的SCYA126基因组序列。序列分析表明,SCYA126基因由2个外显子和1个内含子组成;外显子拼接的序列与SCYA126cDNA序列完全一致,编码92个氨基酸,在N端含有2个相邻的半胱氨酸(CC)和2个不相邻的半胱氨酸,为典型的CC趋化因子亚家族成员;其上游调控序列包含TATA框启动子序列和一些与免疫相关的转录因子结合位点。另外,根据与GenBank接收号为BM029630的EST序列的比较分析,发现SCYA126基因组中的内含子没有被剪接,导致翻译后可能产生N端部分缺失的SCYA126蛋白。在胰脏和肝脏中确认了高表达的包含内含子的mRNA的存在,而且其表达量要明显高于正常剪接的SCYA126mRNA。[中国水产科学,2007,14(1):1-7]     

Pubertal development of male African catfish,Clarias gariepinus. In vitro steroidogenesis by testis and interrenal tissue and plasma levels of sexual steroids

In fish, sex steroids initiate and/or accelerate the maturation of the brain-pituitary-gonad axis. In order to obtain information on the steroid milieu during the pubertal development of male African catfish, we have monitored the conversion of [³H]-pregnenolone and [³H]-androstenedione by testis and [³H]-pregnenolone by interrenal tissue fragmentsin vitro. Pubertal development occurs between two and six months of age. Testicular development proceeds through four main stages that are characterised histologically by the presence of spermatogonia (stage I), spermatogonia and spermatocytes (stage II), spermatogonia, spermatocytes and spermatids (stage III), and all germ cells including spermatozoa (stage IV). 11β-Hydroxyandrostenedione and cortisol were the main products of testes and interrenal tissue, respectively, in all stages of the pubertal development; a change in the steroidogenic pattern was not observed during this period. The interrenal tissue displayed no significant conversion of [³H]-pregnenolone to 11-oxygenated androgens. Blood plasma was analyzed for the presence of five androgens; testosterone, 11β-hydroxytestosterone, 11β-hydroxyandrostenedione, androstenetrione, and 11-ketotestosterone. 11-Ketotestosterone was the quantitatively dominating androgen in the circulation at all stages of development, which was more pronounced in stages III and IV. The obvious differences between thein vitro andin vivo results, namely 11β-hydroxyandrostenedione being the main testicular productvs. 11-ketotestosterone dominating in the blood, may indicate that 11β-hydroxyandrostenedione is converted to 11-ketotestosterone at extratesticular sites.     

Effect of cryoprotectants, extenders and freezing rates on the fertilization rate of frozen striped catfish, Pangasius hypophthalmus (Sauvage), sperm

The effects of four cryoprotectants (methanol, MeOH; dimethyl sulphoxide, DMSO; dimethyl acetamide, DMA; and ethylene glycol, EG), three extenders (calcium‐free Hanks' balanced salt solution, C‐F HBSS, Hanks' balanced salt solution, HBSS and sodium chloride, NaCl) and two different freezing procedures (one‐ and two‐step) on the cryopreservation of striped catfish (Pangasius hypophthalmus (Sauvage)) sperm were investigated. Sperm were frozen using a controlled‐rate freezer in 250 μL straws and stored for 2 weeks in a liquid nitrogen (LN₂) container. They were then airthawed at room temperature, and fertilization, motility and viability were assessed. The highest fertilization rate of 41% (81% of control) was achieved with the combination of 12% DMSO and 0.9% NaCl using a one‐step freezing procedure (10°C min^?1). Also, DMA resulted in a higher fertilization rate (30% or 51% of the control) than MeOH (18% or 38% of the control) or EG (8% or 12% of the control). In addition, the three extenders used did not affect fertilization rates after cryopreservation with each cryoprotectant. There were no significant differences among the three cryoprotectant concentrations and between the one‐ and two‐step freezing procedures. However, fertilization rates of cryopreserved sperm were significantly lower than the controls (P<0.05). The results of this study indicate that high fertilization rates of striped catfish eggs can be achieved using cryopreserved sperm when frozen at 10°C min^?1 in DMSO or DMA with either 0.9% NaCl or C‐F HBSS.     

Inherent variation in growth efficiency of African catfish Clarias gariepinus (Burchell, 1822) juveniles

Understanding the major causes of growth variation is crucial for the success of fish farming since its reduction contributes to maximize production efficiency, reduce food waste and improve water quality. The growth variation observed in aquaculture has been associated with the establishment of social hierarchies. However, some studies suggest that this variation may not be mainly a consequence of social hierarchies but mainly a result of inherent (genetic) differences. This study investigates the magnitude of individual responses, independently of group effects (fish housed individually), in growth efficiency and feeding behaviour of African catfish Clarias gariepinus (Burchell 1822). Despite the low variation in initial body weight (6.5%) and cumulative feed consumption (7.5%) over the experimental period, catfish exhibited high variation in final body weight (18.1%), specific growth rate (17.2%) and feed conversion ratio (27.9%), suggesting that individual variation in growth efficiency is important in determining growth rate. This individual variation may be related with individual differences in protein/fat deposition since faster growing fish deposited more protein and less fat than slower growing fish. Pronounced individual differences in feeding behaviour (reaction towards feed and time spent eating) were also observed and correlated with individual differences in growth efficiency. Fast eaters were the fast growers. We suggest that the growth variation observed in African catfish may be inherent and that the use of grading to increase uniformity should be further investigated.     

Effects of exogenous neurohormone, gonadotropin (GtH) and dopaminergic drugs on the serum GtH content and ovulatory responsiveness of wild catfish, Silurus asotus (Linnaeus, 1758)

Wild female catfish Silurus asotus (Linnaeus, 1758) were injected with domperidone (DOM) alone, [d ‐Ala⁶, Pro⁹ Net]‐luteinizing hormone‐releasing hormone (LHRH‐A) alone once or twice, LHRH‐A plus DOM once or twice simultaneously at 6‐h intervals, LHRH‐A plus carp pituitary extract (CPE) twice simultaneously at 6‐h intervals and LHRH‐A plus human chorionic gonadotropin (HCG) twice simultaneously 6 h apart respectively. The results indicated that injection of LHRH‐A at a dosage of 0.01–0.02 μg g^?1 body weight (BWt) alone induced a low but significant increase in serum gonadotropin (GtH) (P<0.05) and resulted in a very low ovulation rate, while DOM at a dosage of 5 μg g^?1 BWt alone did not induce an increase in the serum GtH levels and ovulation; in contrast, LHRH‐A at a dosage of 0.01 μg g^?1 BWt plus DOM at a dosage of 5 μg g^?1 BWt (termed the Linpe technique) increased the serum GtH (P<0.05) significantly and induced an ovulatory rate of 100%, while LHRH‐A plus CPE or HCG resulted in an increase in the serum GtH (P<0.05) and high ovulatory rate, although the latency period was longer when fish were given LHRH‐A plus HCG or CPE.     

Rearing of African catfish (Clarias gariepinus) and vundu catfish (Heterobranchus longifilis) in traditional fish ponds (whedos): Effect of stocking density on growth, production and body composition

African catfish (Clarias gariepinus) (initial body weight: 34.8 ± 4.8 g) and vundu catfish (Heterobranchus longifilis) (initial body weight: 39.1 ± 8.2 g) fingerlings were stocked at densities of 4, 6 or 8 fish m^− 3 in traditional fish ponds (whedos) constructed in the floodplain of the Oueme River (South Benin, West Africa), for 70 days from March to June 2005. Fish were fed twice a day with 34% crude protein feed formulated with locally available ingredients. The effects of stocking density were evaluated in growth responses, gross production and body composition. Water quality variables were similar (p > 0.05) in all compartments. Temperature and pH were at the optimum level for fish. Dissolved oxygen ranged from 0.9 to 1.2 mg l^− 1 during the experiment and secchi disc transparency was low (< 14 cm). In both species, growth responses increased with the increasing density, significantly in African catfish stocked at density of 8 fish m^− 3 compared to the other densities (4 and 6 fish m^− 3) but not significantly in vundu catfish. Production data ranged from 3.1 ± 0.5 to 22.8 ± 4.5 t ha^− 1 year^− 1 in African catfish and from 6.1 ± 1.2 to 15.1 ± 3.1 t ha^− 1 year^− 1 in vundu catfish. Production increased with increasing stocking densities but only significantly (p < 0.05) between the density of 8 fish m^− 3 and the other densities. In both species, carcass fat increased with increasing density (p < 0.05) while carcass protein and moisture decreased (p > 0.05). These results are important because they indicate that, as far as growth rate and production are concerned, African catfish is more profitable than vundu catfish for culture at high density in whedo.     

Molecular cloning and sequencing of hemoglobin-β gene of channel catfish, Ictalurus punctatus Rafinesque

The hemoglobin-β gene of channel catfish, Ictalurus punctatus, was cloned and sequenced. Total RNA from head kidneys was isolated, reverse transcribed and amplified. The sequence of the channel catfish hemoglobin-β gene consists of 600 nucleotides. Analysis of the nucleotide sequence reveals one open reading frame and 5′- as well as 3′-untranslated regions. The open reading frame of the sequence potentially encodes 148 amino acids with a calculated molecular mass of 16.3 kDa. The pI and charge at pH 7.0 of the deduced hemoglobin-β protein were 7.28 and 0.47, respectively. Overall, 22 amino acid residues were conserved throughout the sequences, including His64 and His93, the sites for heme-binding. Unlike the counterpart of other common cultured fish such as Salmo salar, Oncorhynchus nerka, Oncorhynchus mykiss, Cyprinus carpio and Ctenopharyngodon idella, the hemoglobin-β of channel catfish did not have cysteine. The amino acid sequence of channel catfish hemoglobin-β shows 84% homology with that of Silurus asotus (both are in the order Siluriformes). However, comparison with those of other fish species shows homology ranging from 53 to 68%. Structural analysis by the 3D-PSSM program displays that channel catfish hemoglobin-β has eight α-helices, A–H.     

Electrical stunning followed by decapitation or chilling of African catfish (Clarias gariepinus): assessment of behavioural and neural parameters and product quality

The objective of this study was to assess neural and behavioural responses in farmed African catfish (Clarias gariepinus) upon electrical stunning in combination with decapitation or chilling. To assess the possibility of scaling up one or both experimental methods, two trials were performed in an experimental setting. The product quality of the collected samples was compared with the currently applied industrial method: live chilling. After electrical stunning in combination with decapitation, the fish showed spikes alternated with theta and delta waves on the EEG, followed by minimal brain activity after 20±10 s. The same traces on the EEGs were observed after electrical stunning in combination with chilling. Here, minimal brain activity occurred after 22±11 s. Within a confidence level of 95%, the percentage of African catfish that was effectively stunned after administration of an electrical current of 1.5 A dm⁻², 300 V (50 Hz a.c.), followed by decapitation or chilling was above 91%. The analysis yield and evolution of liquid loss showed significant (P<0.05) differences among the batches, which could be explained by the stunning method used. The course values of the pH in the different batches were significantly (P<0.05) dependent on the stunning method, sex and location (visceral or skin side). It is concluded that African catfish can be stunned effectively using electrical stunning in a water tank, followed by decapitation or chilling in ice. Dutch commercial processors prefer to combine electrical stunning with chilling in flake ice in a rotating tumbler, as the outer slime layer is then removed, which facilitates further processing.     

Dietary levamisole modulates the immune response and disease resistance of Asian catfish Clarias batrachus (Linnaeus)

In order to determine the immunomodulatory effect of dietary levamisole in Asian catfish (Clarias batrachus), fish were fed four different diets for 10 days: a formulated diet as control and the same diet supplemented with 50, 150 or 450 mg levamisole kg^?1 feed. The serum bacterial agglutination titre against Aeromonas hydrophila as a measure of specific immunity, serum haemagglutination titre, natural haemolytic complement activity (ACH₅₀), myeloperoxidase and lysozyme activities, total protein level and oxidative radical production by neutrophils as a measure of non‐specific immunity as well as disease resistance against A. hydrophila challenge to separate vaccinated and non‐vaccinated groups were evaluated at 0, 1, 2 and 3 weeks after last administration of levamisole. Levamisole supplement at the lowest level (50 mg kg^?1) significantly enhanced oxidative radical production and serum myeloperoxidase (MPO) content immediately after 10 days of feeding, which reached peak values after 3 and 2 weeks of feeding respectively. Haemolytic complement and haemagglutination titre were significantly enhanced after 3 and 1 weeks respectively. Haemolytic complement activity and MPO activities were significantly raised to 150 mg kg^?1 after 3 and 2 weeks, respectively. At the highest level of levamisole feeding (450 mg kg^?1) significant decreases in superoxide production and complement activity were measured immediately after levamisole feeding, which returned to the normal level after 1 week post‐ feeding. Fish were challenged with a virulent strain of A. hydrophila at 0, 1, 2 and 3 weeks after levamisole feeding, and the cumulative per cent survival was recorded over 10 days. Feeding levamisole at 50, 150 or 450 mg kg^?1 increased per cent survival in vaccinated fish immediately after levamisole feeding, and survival was significantly higher at 450 mg kg^?1. There was no difference in mortality patterns in non‐vaccinated fish. The results support the use of levamisole at 50 mg kg^?1 feed for 10 days as an immunostimulant in Asian catfish farming.