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Concept: Channel catfish

27

The pectoral spine of catfishes is an antipredator adaptation that can be bound, locked, and rubbed against the cleithrum to produce stridulation sounds. We describe muscle morphology of the pectoral spines and rays in six species in four genera of North American ictalurid catfishes. Since homologies of catfish pectoral muscles have not been universally accepted, we designate them functionally as the spine abductor and adductor and the arrector dorsalis and ventralis. The four muscles of the remaining pectoral rays are the superficial and deep (profundal) abductors and adductors. The large spine abductor and spine adductor are responsible for large amplitude movements, and the smaller arrector dorsalis and arrector ventralis have more specialized functions, that is, spine elevation and depression, respectively, although they also contribute to spine abduction. Three of the four spine muscles were pennate (the abductor and two arrectors), the spine adductor can be pennate or parallel, and ray muscles have parallel fibers. Insertions of pectoral muscles are similar across species, but there is a shift of origins in some muscles, particularly of the superficial abductor of the pectoral rays, which assumes a midline position in Ictalurus and increasingly more lateral placement in Ameiurus (one quarter way out from the midline), and Pylodictis and Noturus (half way out). Coincident with this lateral shift, the attachments of the hypaxial muscle to the ventral girdle become more robust. Comparison with its sister group supports the midline position as basal and lateral migration as derived. The muscles of the pectoral spine are heavier than muscles of the remaining rays in all species but the flathead, supporting the importance of specialized spine functions above typical movement. Further, spine muscles were larger than ray muscles in all species but the flathead catfish, which lives in water with the fastest currents. J. Morphol., 2012. © 2012 Wiley Periodicals, Inc.

Concepts: Catfish, Ictalurus, Blue catfish, Ictaluridae, Channel catfish, Flathead catfish, Fish of North America, Widemouth blindcat

25

Testicular germ cells of channel catfish, Ictalurus punctatus, and blue catfish, I. furcatus were separated into four layers with Percoll density gradient centrifugation, containing different cell types (40 % in the first layer were spermatogonial stem cells, SSCs). Expression of seventeen genes was analyzed for cells from different layers by real-time quantitative PCR. Pfkfb4, Urod, Plzf, Integrin6, IntegrinV, Thy1 and Cdh1 genes showed the same expression change pattern in both channel and blue catfish as these genes were down-regulated in the spermatocytes and even more so in spermatids. Plzf and Integrin6 had especially high expression in SSCs and can be used as SSCs specific markers. Sox2 gene was up-regulated in spermatocytes and even more highly up-regulated in spermatids, which indicated it could be a spermatid marker. In contrast to channel catfish, Id4, Smad5 and Prdm14 gene expressions were strongly down-regulated in spermatocyte cells, but up-regulated in spermatid cells in blue catfish. Smad5 gene was down-regulated in spermatocytes, but up-regulated in both spermatogonia and spermatids, allowing identification as a marker for spermatocytes in blue catfish. Oct4, Id4, Gfrα2, Pum2 and Prdm14 genes showed different expression patterns in the testicular germ cells of channel and blue catfish. This may be a partial explanation to the differing responses of channel catfish and blue catfish to induced spawning technologies. The SSCs specific markers can be used for further SSCs labeling, which can increase the SSCs sorting efficiency and be applied in various studies involving SSCs and other germ cells.

Concepts: DNA, Gene, Cell, Sertoli cell, Catfish, Ictalurus, Ictaluridae, Channel catfish

5

The myostatin (MSTN) gene is important because of its role in regulation of skeletal muscle growth in all vertebrates. In this study, CRISPR/Cas9 was utilized to successfully target the channel catfish, Ictalurus punctatus, muscle suppressor gene MSTN. CRISPR/Cas9 induced high rates (88-100%) of mutagenesis in the target protein-encoding sites of MSTN. MSTN-edited fry had more muscle cells (p < 0.001) than controls, and the mean body weight of gene-edited fry increased by 29.7%. The nucleic acid alignment of the mutated sequences against the wild-type sequence revealed multiple insertions and deletions. These results demonstrate that CRISPR/Cas9 is a highly efficient tool for editing the channel catfish genome, and opens ways for facilitating channel catfish genetic enhancement and functional genomics. This approach may produce growth-enhanced channel catfish and increase productivity.

Concepts: DNA, Gene, Genetics, Genomics, Catfish, Ictalurus, Ictaluridae, Channel catfish

3

The channel catfish (Ictalurus punctatus), a species native to North America, is one of the most important commercial freshwater fish in the world, especially in the United States' aquaculture industry. Since its introduction into China in 1984, both cultivation area and yield of this species have been dramatically increased such that China is now the leading producer of channel catfish. To aid genomic research in this species, data sets such as genetic linkage groups, long-insert libraries, physical maps, bacterial artificial clones (BAC) end sequences (BES), transcriptome assemblies, and reference genome sequences have been generated. Here, using diverse assembly methods, we provide a comparable high-quality genome assembly for a channel catfish from a breeding stock inbred in China for more than three generations, which was originally imported to China from North America.

Concepts: Gene, Genetics, Fish, United States, Catfish, Ictalurus, Ictaluridae, Channel catfish

2

Aquaculture recently overtook capture fisheries as the largest producer of food fish, but to continue increasing fish production the industry is in search of better methods of improving fish health and growth. Pre- and probiotic supplementation has gained attention as a means of solving these issues, however, for such approaches to be successful, we must first gain a more holistic understanding of the factors influencing the microbial communities present in the intestines of fish. In this study, we characterize the bacterial communities associated with the digestive tract of a highly valuable U.S. aquaculture species, channel catfish Ictalurus punctatus, over the first 193 days of life to evaluate temporal changes that may occur throughout ontogenetic development of the host. Intestinal microbiota were surveyed with high-throughput DNA sequencing of 16S rRNA V4 gene amplicons derived from fish at 3, 65, 125, and 193 days post hatch (dph), while also characterizing the environmental microbes derived from the water supply and the administered diets. Microbial communities inhabiting the intestines of catfish early in life were dynamic, with significant shifts occurring up to 125 dph when the microbiota somewhat stabilized, as shifts were less apparent between 125 to 193 dph. Bacterial phyla present in the gut of catfish throughout ontogeny include Bacteroidetes, Firmicutes, Fusobacteria, and Proteobacteria; with the species Cetobacterium somerae and Plesiomonas shigelloides showing the highest abundance in the catfish microbiota after 3 dph. Comparisons of the gut microbiota to the environmental microbes reveals that the fish gut is maintained as a niche habitat, separate from the overall microbial communities present in diets and water-supply. Although, there is also evidence that the environmental microbiota serves as an inoculum to the fish gut. Our results have implications for future research related to channel catfish biology and culture, and increase our understanding of ontogenetic effects on the microbiota of teleost fish.

Concepts: Archaea, Bacteria, Gut flora, Microbiology, 16S ribosomal RNA, Aquaculture, Catfish, Channel catfish

0

Repressible knockdown approaches were investigated to manipulate for transgenic sterilization in channel catfish, Ictalurus punctatus. Two primordial germ cell (PGC) marker genes, nanos and dead end, were targeted for knockdown and an off-target gene, vasa, was monitored. Two potentially copper-sensitive repressible promoters, yeast ctr3 (M) and ctr3-reduced (Mctr), were coupled with four knockdown strategies separately including: ds-sh RNA targeting the 5' end (N1) or 3' end (N2) of channel catfish nanos, full-length cDNA sequence of channel catfish nanos for overexpression (cDNA), and ds-sh RNA-targeting channel catfish dead end (DND). Each construct had an untreated group and treated group with copper sulfate as the repressor compound. Spawning rates of full-sibling P1 fish exposed or not exposed to the constructs as treated and untreated embryos were 85 and 54%, respectively, indicating potential sterilization of fish and repression of the constructs. In F1 fish, mRNA expressions of PGC marker genes for most constructs were downregulated in the untreated group and the knockdown was repressed in the treated group. Gonad development in transgenic, untreated F1 channel catfish was reduced compared to non-transgenic fish for MctrN2, MN1, MN2, and MDND. For 3-year-old adults, gonad size in the transgenic untreated group was 93.4% smaller than the non-transgenic group for females and 92.3% for males. However, mean body weight of transgenic females (781.8 g) and males (883.8 g) was smaller than of non-transgenic counterparts (984.2 and 1254.3 g) at 3 years of age, a 25.8 and 41.9% difference for females and males, respectively. The results indicate that repressible transgenic sterilization is feasible for reproductive control of fish, but negative pleiotropic effects can result.

Concepts: DNA, Gene, Gene expression, RNA, Germ cell, Catfish, Ictalurus, Channel catfish

0

Until recently, use of antibiotics to enhance terrestrial animal growth performance was a common, U.S. Food and Drug Administration-approved, but controversial practice. There are no FDA-approved production claims for antibiotic drug use in fish, but it is a common misconception that antibiotics are widely used for this purpose in U.S. aquaculture. Antibiotics are not thought to be effective growth promoters in fish, but there is little quantitative data available to address whether there are growth-promoting effects that might incentivize use of antibiotics in this way, despite legal prohibitions. Therefore, this study was conducted to determine if oral administration of oxytetracycline, an antibiotic with known growth-promoting effects in terrestrial livestock, has a similar effect when applied to Channel Catfish Ictalurus punctatus, hybrid Striped Bass Morone chrysops × M. saxatilis, Nile Tilapia Oreochromis niloticus, or Rainbow Trout Oncorhynchus mykiss. Oxytetracycline products with production claims are typically applied at doses substantially lower than the approved therapeutic doses for the same products. Medication (0, 0.24 or 1.2 g oxytetracycline dihydrate/kg feed) and feeding rates (3% body weight/d) were selected to achieve target doses of 0, 16, or 80 mg/kg fish/d representing control, sub-therapeutic, and therapeutic treatments. Replicate groups of fish (N = 4) were fed accordingly for 8 weeks. Overall, oral administration of oxytetracycline did not affect survival or promote growth of the selected taxa, with no significant differences observed for weight gain, feed conversion ratio, or specific growth rate (P > 0.05 in all cases). Few differences were observed in organosomatic indices and in the frequency of tissue abnormalities; where present, these differences tended to suggest a negative effect of long-term dietary exposure to oxytetracycline. These data demonstrate that there is no benefit to dietary supplementation with oxytetracycline for nontherapeutic purposes in a range of economically important finfish species. As such, our results indicate there is little incentive to misuse oxytetracycline products for purposes of growth promotion in U.S. aquaculture.

Concepts: Cichlid, Salmon, Tilapia, Oreochromis, Oreochromis niloticus, Catfish, Channel catfish, Striped bass

0

Channel catfish skin is a by-product from catfish fillet production. Collagens were extracted from catfish skins by: (1) acid; (2) homogenization-aided; and (3) pepsin-aided extraction methods. Kinetic analysis of extraction was performed. SDS-PAGE was carried out for all collagens extracted under different conditions. Protein solubility, zeta potential, circular dichroism and gel strength of the collagen extracted by three methods were studied to determine optimal extraction conditions. Protein recovery rate from minced skins extracted with pH 2.4 HCl containing 23.6KU/g pepsin was the highest (64.19%). SDS-PAGE showed that collagens extracted with different methods had different proteins ratio patterns, even though the molecular mass of collagen subunits were similar, 123 and 113KDa for α1and α2chains, 226KDa for β chain and 338.5KDa for γ chain, respectively. Channel catfish skin collagens were typical type I collagens and could have applications in food, medical and cosmetic industries.

Concepts: Protein, Molecular biology, Collagen, Keratin, Catfish, Ictalurus, Ictaluridae, Channel catfish

0

The full-length complementary DNA of two genes related to vertebrate albinism, the tyrosinase gene tyr and tyrosinase-related protein 1 gene tyrp1, were cloned and analysed from normal and albino yellow catfish Tachysurus fulvidraco. The open reading frames (ORF) of tyr and tyrp1 encode putative peptides of 533 and 526 amino acids (amino-acid), both of which possess two conserved copper binding sites. The homologous identities of deduced amino-acid sequences showed that both Tyr and Tyrp1 of T. fulvidraco share considerable similarity with that of channel catfish Ictalurus punctatus. Both tyr and tyrp1 were expressed in a wide range of adult tissues. Tyr gene had the highest expression level in the brain of both normal and albino T. fulvidraco. Tyrp1 had the highest expression level in the skin of normal groups, and the fin of albino groups. The messenger (m)RNA expressions of tyr and tyrp1 were detectable at different early developmental stages and varied with embryonic and larval growth. Tyr and tyrp1 mRNA have obvious tissue specificity both in normal and albino T. fulvidraco and higher expression levels were detected in the normal group revealing that tyr and tyrp1 may have an important role in pigmentation. These results will provide useful data for understanding the molecular mechanism of melanin formation and the occurrence of albinism in T. fulvidraco.

Concepts: DNA, Protein, Gene, Genetics, Gene expression, Amino acid, Molecular biology, Channel catfish

0

The objective of this work was to extract gelatin from skin of channel catfish (Ictalurus punctatus) and to study its functional properties and application in ice-cream and beer. SDS-PAGE patterns showed that channel catfish gelatin (CCG) had more high molecular weight components (β and γ chains) than had calf bone gelatin (CBG). The viscosity of CCG was 42mPa.s at 10°C, which was three times more than that of CBG. Compared to CBG, CCG presented higher emulsion capacity and stability, as well as higher foaming stability. CCG conferred the ice-cream samples and beer with better mouth feel and clarification effect, respectively. The results indicated that CCG had great potential to be utilized in the food industry.

Concepts: Catfish, Ictalurus, Ictaluridae, Channel catfish