鲤基因组中1个新Tc1类转座子的发现与鉴定

在鲤(Cyprinus carpio)基因组中鉴别出一个新的具有潜在转座活性的Tc1类转座子,并命名为CCTN转座子(Cyprinus carpio Transposon,CCTN)。CCTN转座子全长1 611 bp,由两端约214 bp的反向重复序列(Inverted Repeat,IR)和中间不间断的996 bp的转座酶开放阅读框(Open Reading Frame,ORF)组成。CCTN转座子推测的转座酶序列中存在完整的DD(34)E结构域,此结构域是Tc1类转座酶作用的必需位点之一。采用实时荧光定量PCR评估CCTN转座子在鲤基因组中的拷贝数约为2.28×103,占全基因组的0.21%。分子系统学研究表明,CCTN转座子是一个新的鱼类Tc1类转座子,其与斑马鱼(Danio rerio)Tzf-28、大西洋鲑(Salmo salar)SALT1和鲽(Pleuronectes platessa)PPTN2等Tc1类转座子亲缘关系较近。     

刀鲚类Tc1转座子的分子特征及拷贝数变化的意义

为了探讨刀鲚2种不同生活史种群的遗传结构差异以及成因,本研究利用分子克隆及转座子展示技术,从刀鲚基因组中分离、鉴定出一类新的、命名为Cn-Tc1的转座子。该转座子全长1896 bp,为鳀科鱼类第一类被挖掘的Tc1转座子。Cn-Tc1自身包含另一个长度为1040 bp的类Tc1,表明Cn-Tc1在基因组内的转座经历过多次迸发。Cn-Tc1的5’和3’末端反向重复序列长度分别为64和83 bp,转座插入位点具有"TATA"基序。预测的Cn-Tc1转座酶具有与DNA结合的保守结构,提示其仍具有转座潜能。Cn-Tc1插入位点侧翼序列的GC含量呈不均匀分布,均值低于AT含量。运用荧光定量PCR方法,估算了靖江、象山、洞庭湖、鄱阳湖、太湖以及崇明等水域刀鲚种群基因组中Cn-Tc1拷贝数,分别为3.140×103、2.992×103、6.876×103、5.205×103、5.531×103和3.046×103个。单因素方差分析发现,象山、崇明、靖江种群间的拷贝数差异性不显著,而与其他种群的差异性均显著;鄱阳湖、太湖和洞庭湖种群之间差异性亦不显著,但与其他种群均呈显著性差异。研究结果表明,Cn-Tc1促进了遗传结构的改变,为刀鲚种群的适应性进化提供了自然选择的基础。     

Folate-Degrading Bacteria in Channel Catfish Feeds

Two "Pseudomonas" spp., designated as PF1 and PF2, were isolated from 147 of 148 channel catfish feed samples and 10 different orgns from anemic and healthy channel catfish, "Ictalurus punctatus." Both PF1 and PF2 are capable of using folic acid as the sole source of carbon and nitrogen, and they hydrolized folic acid to pteroic acid. They did not utilize common carbohydrates. In vitro, the bacteria destroyed over 93% of folic acid in media containing only folic acid and inorganic salts. The morphological and biochemical properties of the bacteria indicate that they are "Pseudomonas" spp. and similar to some other "Pseudomonas" spp. and similar to some other "Pseudomonas" species that utilize folic acid. It is suspected that these folate degrading bacteria are involved in a nutritionally related anemia of cultured channel catfish.     

Early Growth and Morphology Among Hybrids of Ictalurid Catfishes

Half-sib ictalurid families were produced when eggs from three channel catfish, "Ictalurus punctatus," females were fertilized with a mixture of sperm from channel catfish, blue catfish, "I. furcatus," black bullhead, "Ameiurus melas," and flathead catfish, "Plyodictis olivaris." Sperm from all four species successfully fertilized channel catfish eggs, although individual families contained different percentages of the various crosses. The enzyme glucose phosphate isomerase distinguished the different hybrids and parental groups, and confirmed sorting of offspring based on morphology. At 1 month of age, channel catfish ♀ X flathead catfish ♂ hybrids (0.36 ± 0.07 g, mean ± SD) were heavier (P ≤ 0.05) than channel catfish ♀ X black bullhead ♂ (0.12 ± 0.03 g), channel catfish ♀ X blue catfish ♂ hybrids (0.11 ± 0.03 g ), and channel catfish (0.10 ± 0.4 g). By 8 months of age, channel catfish ♀ X black bullhead ♂ hybrids (36.4 ± 21.9 g) and channel catfish ♀ X blue catfish ♂ hybrids (34.5 ± 24.2 g) were similar in weight, and each was significantly heavier than channel catfish ♀ X channel catfish ♂ (26.9 ± 16.1 g), while channel catfish ♀ X flathead catfish ♂ (17.0 ? 6.8 g) weighed the least. Morphometric ratios of channel catfish ♀ X blue catfish ♂ were intermediate between channel catfish ♀ X channel catfish ♂ and blue catfish for four of 16 features and were similar to either or both of the parental species for 10 others. Ratios for channel catfish ♀ X black bullhead ♂ hybrids were intermediate between channel catfish ♀ X channel catfish ♂ and black bullhead for seven of 16 features. Ratios for channel catfish ♀ X flathead catfish ♂ hybrids were larger than channel catfish ♀ X channel catfish ♀ for eight of 16 features and smaller than channel catfish ♀ X channel catfish ♂ for five of 16 features. All channel catfish ♀ X flathead catfish ♂ hybrid offspring were female, based on examination of the urogenital region or gross appearance of the gonands, while other groups had normal sex ratios. Production of half-sib interspecific hybrid families provides a unique opportunity to evaluate genetic influences on commercially important traits and to evaluate the potential of these ictalurid hybrids in aquaculture.     

水库网箱和池塘养殖斑点叉尾（鱼回）肌肉营养成分和品质的比较分析

为了了解池塘和水库网箱两种养殖模式下斑点叉尾鮰（Ictalurus punctatus）肌肉营养成分和品质特性,利用生化分析、物性分析方法和电感耦合等离子体原子发射光谱法分别对斑点叉尾鮰肌肉的常规营养成分、系水力、质构特性及肌肉矿物元素含量进行了测定。结果显示,池塘和水库网箱养殖的斑点叉尾鮰含肉率都在65%以上;池塘组的贮存损失和失水率显著低于水库网箱养殖组,而冷冻渗出率显著高于水库网箱养殖组;水库网箱养殖组的粗蛋白、粗灰分含量均显著高于池塘组,而粗脂肪含量显著低于池塘组;两组中矿物元素含量都比较丰富,水库网箱养殖组肌肉中钾、磷、钙、镁、锌、铁六种元素的含量均显著高于池塘组;池塘组肌肉的凝聚性和回复性显著高于水库网箱养殖组,而肌肉硬度、胶黏性和咀嚼性显著低于水库网箱养殖组。研究表明,池塘养殖的斑点叉尾鮰肌肉具有较好的系水力,而水库网箱养殖的斑点叉尾鮰肌肉具有高蛋白低脂肪、矿物元素更丰富、肉质硬度大的特点。     

Integrated application of two different screening strategies to select potential probiotics from the gut of channel catfish Ictalurus punctatus

In this study, 342 bacterial strains were isolated from the gut of channel catfish Ictalurus punctatus. Two different screening strategies, including the production of antagonistic effects against pathogens and the capacity of secreting exoenzyme, were used to select potential probiotics from the bacterial strains. Among them, 6 isolates showed antagonism against the pathogens, 15 isolates could produce high amylase and protease, and 3 isolates (BHI344, BH0142 and BH03124) had both the two characteristics . Then, BHI344, BH0142, and BH03124 were tested for survival rate in channel catfish bile and at low pH, antimicrobial susceptibilities, and pathogenicity in channel catfish. The results of the obtained assays showed that BHI344 possessed the broadest inhibitory potential, a strong ability of secreting extra-cellular amylase and protease, high viability in simulated gastrointestinal conditions, and sensitivity to most of the used antibiotics and non-pathogenic to channel catfish. Therefore, it could be selected as the most promising probiotics candidate. BHI344 was identified as Bacillus subtilis based on the phenotypic characterization and 16S rRNA gene sequencing.     

Organization and sequence of four flagellin-encoding genes of Edwardsiella ictaluri

Edwardsiella ictaluri , the cause of enteric septicaemia in channel catfish ( Ictalurus punctatus ), is motile by means of peritrichous flagella. We determined the complete flagellin gene sequences and their organization in E. ictaluri by sequencing genomic segments from a λ-ZAP phage genomic library of E. ictaluri . Four flagellin genes ( fliC1, fliC2, fliC3 and fliC4 ) are arranged in tandem within 6 kb in the E. ictaluri genome. Each flagellin-coding sequence is preceded by a σ²⁸ recognition site consensus sequence. The predicted amino acid sequences of all four flagellin proteins (between 36 and 37.5 kDa) are similar in the N-terminal (1–160 aa) and C-terminal (last 74 aa) portions and are divergent in the central portion of the proteins. Proteins encoded by flC1, fliC2 and fliC3 are more similar to each other (88–90% aa identity) than to the protein encoded by fliC4 (76–78% aa identity). basic local alignment search tool analysis of GenBank sequences showed that all flagellin aa sequences are more similar to those of Serratia marcescens (72–74% identity) than to those of Edwardsiella tarda (≤64% identity). Primary determination of E. ictaluri flagellin gene sequences facilitate advanced studies on the role of flagella in host–pathogen interaction.     

Isolation of a novel hepatic CYP2-related protein from channel catfish, Ictalurus punctatus

Channel catfish (Ictalurus punctatus) have been previously shown to express two major cytochrome P450 (CYP) protein bands that are cross-reactive with anti-CYP2K1 (rainbow trout, Oncorhynchus mykiss) antibodies on Western blots. These proteins appear to be the major constitutive CYPs in channel catfish and show distinct sex- and age-specific variations in expression. Because I. punctatus is an important agricultural and ecological commodity, and because it displays a high degree of resistance to the toxic effects of many pesticides, the molecular and catalytic characteristics of its biotransformation systems are of interest to those in areas of environmental science and aquaculture research. Using a chromatographic method similar to that employed in the purification of other fish CYP2 enzymes, a single CYP2-related protein (CM-HA3) was isolated from channel catfish hepatic microsomes. The isolated protein displays a relative molecular mass of approximately 47 kDa, and a CO-reduced difference spectrum _max of 449.6 nm. The sequence of 15 residues at the amino-terminal of CM-HA3 is 27% identical to both CYP2K1 and CYP2M1 isoforms of rainbow trout. Correlational analysis was employed to characterize potential substrates for this isoform, but no significant relationship was observed with E₂ hydroxylation, testosterone hydroxylation, or 7-ethoxycoumarin O-deethylase activities. These data indicate that CM-HA3 is a CYP2 family protein, with as yet uncharacterized substrate specificities.     

Culture environment and the odorous volatile compounds present in pond-raised channel catfish (<Emphasis Type="Italic">Ictalurus punctatus</Emphasis>)

A headspace solid-phase micro-extraction (SPME) coupled with GS-MS method was used to measure volatile compounds in fillets from musty off-flavor, muddy off-flavor, and on-flavor channel catfish (Ictalurus punctatus), along with water and soil samples from the farm ponds in which the fish had been raised. Two ponds of each type of flavor were selected, and five fish, water, and soil samples were collected from each pond. Linear and multiple linear regression analyses were carried out between/among off-flavor strength and volatile compound contents to investigate their possible correlations. The combination of two strong off-flavor compounds, 2-methylisoborneol (MIB) and geosmin (GSM), was probably mainly responsible for the musty off-flavor in the catfish fillets, and an odorous alcohol, 1-hexanol, was correlated with muddy off-flavor (p =?0.015). There was a strong correlation between beta-cyclocitral and MIB in a pond that gave musty off-flavor catfish contents (p =?0.006), suggesting that these compounds might be generated by similar cyanobacteria. The contents of GSM, MIB, and beta-cyclocitral were high in the water of ponds that yielded off-flavor fish, indicating that catfish might acquire these compounds from pond water.     

Effects of Environmental Sodium Chloride on Percent Hatch, Yolk Utilization, and Survival of Channel Catfish Fry

Abstract.— Only limited research has addressed the effect of salinity on hatching of channel catfish Ictalurus punctatus eggs, and no studies have evaluated the effect of salinity on fry development and survival. This study was undertaken to determine the effect of environmental sodium chloride (0, 1, 2, and 4 g/L. NaCl) on percent hatch, yolk utilization, and survival of channel catfish fry. Experiments were conducted in recirculating systems using seven egg masses (1–2 d old). Each egg mass was divided into smaller portions which remained undissociated or were dissociated with sodium sulfite (NaSO₃). Eggs were incubated until hatching. Wet and dry weights were obtained for sacfry at 1 and 5 d post-hatch to determine wet weight gain and dry weight loss, and fry were sampled 7 d after initiation of exogenous feeding to determine survival. Percent hatch, yolk utilization, and survival of fry hatched from undissociated eggs were greatest at 1 g/L NaCl. In addition, treatment of eggs with NaSO₃ significantly reduced percent hatch at all NaCl levels. Although our results indicate that addition of NaCI to hatchery water supplies can increase production of channel catfish fry, additional research is needed before this practice can be recommended on a commercial basis.