Assignment of immune-related genes to the channel catfish, Ictalurus punctatus, genetic map

Eighteen new genes, adenosine A1 receptor (ADORA1), complement component 4-beta (C4b), complement component 8-beta (C8b), chemokine ligand 19 (CCL19), chemokine ligand 21 (CCL21), chemokine ligand 25 (CCL25), chemokine receptor 2 (CCR2), chemokine receptor 5 (CCR5), chemokine receptor 4 (CCR4), chemokine receptor 7 (CCR7), chemokine receptor 9 (CCR9), interleukin 1-beta (IL1B), integrin II-beta (ITGB2), novel immune type receptor 2 (NITR2), novel immune type receptor 4 (NITR4), natural killer cell lysin (NKLYSIN), nucleotide excision repair (RAD23B) and tumour necrosis factor-alpha (TNF), were assigned to the channel catfish (Ictalurus punctatus) genetic linkage map. Polymorphic microsatellite markers were developed for NITR2, NITR4 and RAD23B from short-tandem repeats in the available sequence. Polymorphic microsatellite markers were developed for the remaining 15 genes by short-tandem repeat-anchored primer sequencing of catfish bacterial artificial chromosomes. Two gene clusters (MYOG-NRAMP-ADORA1) and (CCR4-CCR2-CCR5) displayed conservation of synteny between catfish and mammals. Assignment of 18 new genes to the catfish linkage map will further advance integration of genetic and physical maps and comparative mapping between channel catfish and map rich species.     

An AFLP-based genetic linkage map of channel catfish (Ictalurus punctatus) constructed by using an interspecific hybrid resource family

Catfish is the major aquaculture species in the United States. The hybrid catfish produced by crossing channel catfish females with blue catfish males exhibit a number of desirable production traits, but their mass production has been difficult. To introduce desirable genes from blue catfish into channel catfish through introgression, a genetic linkage map is helpful. In this project, a genetic linkage map was constructed using amplified fragment length polymorphism (AFLP). A total of 607 AFLP markers were analyzed using 65 primer combinations and an interspecific backcross resource family. A total of 418 AFLP markers were assigned to 44 linkage groups. Among the remaining 189 markers, 101 were not used because of significant segregation distortion, 29 were unlinked, and 59 were eliminated because they span very large distances. The 418 AFLP markers covered 1593 cM Kosambi. The AFLP markers showed a high level of clustering that appears to be related to certain primer combinations. This linkage map will serve as the basis for mapping a greater number of markers to provide a map with high enough resolution for it to be useful for selective breeding programs using introgression.     

Density-independent growth of floodplain river channel catfish Ictalurus punctatus

This study investigated the relative influence of biotic and abiotic factors on the growth of channel catfish Ictalurus punctatus in seven Mississippi floodplain rivers. The results indicate that growth was density-independent, being defined largely by abiotic conditions.     

Organization of the telencephalon in the channel catfish,Ictalurus punctatus

The cytoarchitectonics of the telencephalon of the channel catfish, Ictalurus punctatus, are described as a basis for experimental analysis of telencephalic afferents and efferents. The olfactory bulb comprises: (1) an outer layer of olfactory nerve fibers, (2) a glomerular layer, (3) an external cell layer, (4) an inner fiber layer, and (5) an internal cell layer. The telencephalic hemispheres comprise the areas ventralis and dorsalis telencephali. The area ventralis consists of: (1) a precommissural, periventricular zone including nucleus 'nother (Vn), the ventral nucleus (Vv), and the dorsal nucleus (Vd); (2) a precommissural, migrated zone of central (Vc) and lateral (VI) nuclei; (3) a supracommissural nucleus (Vs); (4) a caudal commissural zone of postcommissural (Vp) and intermediate (Vi) nuclei; and (5) a preoptic area (PP). The area dorsalis comprises: (1) medial (DM), (2) dorsal (Dd), (3) lateral [DL, containing dorsal (DLd), ventral (DLv), and posterior (DLp) regions], (4) posterior (DP), and (5) central (DC-1, -2, -3) areas. Nucleus taeniae (NT) is transitional between areas dorsalis and ventralis.     

Histology of cultured channel catfish, Ictalurus punctatus (Rafinesque)

Dissolved oxygen and un-ionized ammonia concentrations were monitored in 12 0.04 ha earthen ponds stocked with 10 000 channel catfish, Ictalurus punctatus ,/ha. Gill, liver and trunk kidney tissue samples were removed periodically for histological examination. Total ammonia and dissolved oxygen levels were in the ranges reported for catfish culture at this level of intensity. Average un-ionized ammonia nitrogen concentrations ranged from 20 to 67 μgh⁻¹ and average daily maxima ranged from 63 to 183 μgh⁻¹. Gill lesions characteristic of un-ionized ammonia exposure were common in fish from all ponds.     

Gastrointestinal torsion in a channel catfish (Ictalurus punctatus)

A case of gastrointestinal torsion with dilatation in a farm-raised channel catfish (Ictalurus punctatus) was examined at the Thad Cochran National Warmwater Aquaculture Center (Stoneville, Mississippi, USA). The affected fish was a gravid female broodfish, which displayed pale gills and a markedly distended abdomen. Internal examination revealed that the gastrointestinal tract and ovaries were rotated around each other four times in a counterclockwise direction as viewed in right lateral recumbency. The catfish had a markedly distended gastrointestinal tract, pale liver, hypoplastic spleen, hypoplastic swim bladder, and high volume of ascitic fluid. Blood analysis indicated multiple abnormalities, including severe anemia and metabolic acidosis. The etiology of the torsion was uncertain; however, the presence of a hypoplastic swim bladder most likely allowed for increased movement of the gastrointestinal tract and ovaries. When examining cases of abdominal distention in fish, gastrointestinal torsion can be considered among the differential diagnoses.     

A high-affinity zinc-binding plasma protein in channel catfish (Ictalurus punctatus).

1. Channel catfish (Ictalurus punctatus) have a remarkably high concentration of zinc (Zn) in their blood serum, about 20 micrograms/ml. However, compared to mammals, the concentrations of Zn in their tissues are not remarkable. The serum Zn is dialyzable against a solution containing 1 mM EDTA. 2. Following separation of serum proteins by gel-filtration most of the Zn was recovered in a fraction with the same peak volume of elution for the Zn and protein concentrations and having a molecular weight similar to bovine serum albumin. 3. Binding of Zn to such sites was not changed by Cu2+, Cd2+, Ca2+, or La3+. N-ethylemaleimide (NEM) appeared to decrease binding slightly. 4. Equilibrium dialysis with a Scatchard plot analysis of these data suggested that a single set of binding sites was present on the protein(s) with KD of 2 x 10(-5) M. There were binding sites for 35 x 10(-8) M Zn/mg protein. 5. Parallel equilibrium dialysis measurements using human, rabbit and chicken albumins indicated that the catfish serum protein(s) had a much higher affinity and binding capacity for Zn than the albumins in these species. 6. The catfish Zn serum-binding protein may be an albumin. The possible physiological significance of such a serum protein in freshwater fish is discussed.     

Environmental stress and bacterial infection in channel catfish, Ictalurus punctatus Rafinesque

Channel catfish, Ictalurus punctatus , were injected intraperitoneally with a sublethal dose of Aerornonas hydrophila and then stressed for 144 h by being maintained either in a dissolved oxygen concentration of 1·5 mg/1, 1·2 mg/1 total ammonia, and/or 6·5 mg/1 free CO₂ with a continuous inflow of water. A significant difference in percentage of mortality was noted between treatments ( P < 0·05). The trunk kidneys of surviving stressed fish had significantly higher total bacterial counts than non-stressed controls. A. hydrophila was isolated from 67% of the stressed fish and 9% of the control fish. Edwardsiella tarda , apparently endemic in the population, was isolated from 43% of the stressed fish and 7% of the control fish. Histopathological lesions were in the gills, liver, spleen, trunk kidney, and head kidney of stressed fish, but not control fish.     

Cloning and characterization of microsatellite loci in channel catfish, Ictalurus punctatus

Short tandem repeat (microsatellite) loci were cloned from the channel catfish, Ictalurus punctatus , genome for use as molecular markers for genetic improvement of this important agricultural species. Plasmid clones containing catfish genomic DNA inserts were identified, by hybridization with tandem repeat DNA probes, and sequenced using automated laser fluorescence. A feral population of catfish displayed levels of heterozygosity greater than 0·7 for 13 of 22 loci and heterozygosity greater than 0·5 for 20 of 22 loci. Allelic polymorphism ranged from three to 17 alleles per locus in the feral population. Populations of domestic, farm-raised catfish and a research strain displayed levels of heterozygosity similar to the feral population. Non-invasive tissue sampling provided abundant material for the polymerase chain reaction-based genotype assay. The microsatellite loci will be useful in the molecular characterization and genetic improvement of channel catfish populations.     

Structure and expression of transferrin gene of channel catfish,Ictalurus punctatus

Transferrin is important in iron metabolism and has been reported to be involved in disease defence responses after bacterial infection. In this study, we identified, sequenced, and characterized the transferrin gene from channel catfish, Ictalurus punctatus. The catfish transferrin gene was similar to those of other vertebrate species with 17 exons and 16 introns. Sequence analysis indicated the presence of the two duplicated lobes, each containing two sub-domains separated by a cleft harboring the iron-binding site, suggesting their structural conservation. The channel catfish transferrin cDNA encodes 679 amino acids with 42–56% similarity to known transferrin genes from various species. Southern blot analysis suggested the presence of two copies of the transferrin gene in the catfish genome, perhaps arranged in a tandem fashion. The catfish transferrin gene was mapped to a catfish BAC-based physical map. The catfish transferrin gene was highly expressed in the liver, but expression was low in most other tested tissues. Transferrin expression was significantly up-regulated after infection with Edwardsiella ictaluri, the causative agent of enteric septicemia of catfish. Such induction was also found with co-injection of iron-dextran and E. ictaluri, while transferrin expression was not significantly induced with the injection of iron-dextran alone.     

A study of haematological profiles of channel catfish, Ictalurus punctatus (Rafinesque)

A blood profile of Ictalurus punctatus was determined from a total of 35 fish. The values obtained included plasma protein, haemoglobin, red cell counts and differential white cell counts.     

Immune hypersensitivity in the channel catfish, Ictalurus punctatus (Rafinesque)

Channel catfish, Ictalurus punctatus , exhibited passive cutaneous anaphylaxis (PCA) when sensitized with Flexibacter columnaris antiserum produced in channel catfish (the first record of PCA in any fish), but not when sensitized with homologous antiserum against channel catfish virus. Channel catfish did not give delayed skin hypersensitivity responses after immunization with either channel catfish virus or F. columnaris .     

Hepatic drug metabolizing enzyme activity in the channel catfish, Ictalurus punctatus

1. Several pathways of drug metabolizing enzymic activity were measured in hepatic fractions of the channel catfish and rat using model substrates. The pathways examined included the O-demethylation of p-nitroanisole, microsomal ester hydrolysis of procaine and glucuronidation of p-nitrophenol, and the cytosolic acetylation of sulfamethazine and sulfation of 2-naphthol. Catfish liver preparations were incubated at both 25 degrees C and 37 degrees C. 2. The oxidative metabolism of p-nitrophenol was only one-eighth that of the rat at 37 degrees C and one-twelfth that of the rat at 25 degrees C. 3. Procaine ester hydrolysis was negligible in catfish microsomal preparations. 4. At 37 degrees C, p-nitrophenol glucuronidation was equivalent in catfish and rat microsomes. 5. Catfish cytosolic preparations exhibited N-acetyltransferase and arylsulfotransferase nearly comparable to those of the rat. 6. Rates of glucuronidation and sulfation were higher at 37 degrees C than at 25 degrees C in hepatic fractions of catfish.     

A standardized microsatellite marker panel for parentage and kinship analyses in channel catfish,Ictalurus punctatus

This research was designed to produce a standardized set of microsatellite loci for parentage and kinship analyses in channel catfish, the leading species of US aquaculture. Three panels of five to six markers each were developed that contained a total of two dinucleotide‐, eight trinucleotide‐ and seven tetranucleotide‐microsatellite loci respectively. The loci had a range of nine to 31 alleles per locus in an outbred population. Based on the allele frequencies measured in commercial randomly bred broodstock, the combined probability of non‐exclusion of an unrelated candidate parent pair was 5.36e‐18. The combined probability of non‐exclusion of unrelated identical genotypes was 2.58e‐08. The microsatellite panels were validated by parentage and kinship evaluation in three populations. A total of 697 spawns were collected from matings of outbred broodstock over three spawning seasons, and parents were determined unambiguously for all but three spawns. Genotype analysis also enabled the identification of half‐sibling and full‐sibling families produced by pond spawning. In a second experiment, parentage was unambiguously determined in nine spawns from a population consisting of broodstock derived from only four families. A third experiment demonstrated that all but one of 374 individuals from 10 full‐sibling families could be assigned to a family after coculture in an earthen pond for 1 year. The standardized microsatellite panels enable the development of pedigreed catfish populations and large‐scale performance evaluations in common environments to support the genetic improvement of cultured catfish through selective breeding.     

Histophathology and electron microscopy of channel catfish virus in infected channel catfish,Ictalurus punctatus (Rafinesque)*

A histologic and electron microscopic study was made on selected organs from channel catfish Ictalurus punctatus (Rafinesque) fingerlings that were experimentally infected with channel catfish virus (CCV). Histopathology was characterized by necrosis and haemorrhage in kidney and liver, and haemorrhage in the spleen and gastrointestinal tract. Virus replication occurred in nuclei of cells in the kidney, liver and spleen. Intranuclear inclusion bodies consisting of geometric crystalline arrays and lamellar structures were associated with virus replication.     

Fine structure of the nasal barbel of the channel catfish,Ictalurus punctatus

Ultrathin sections of the nasal barbel of the channel catfish, Ictalurus punctatus, were studied in the electron microscope and the fine structure was compared to that of barbels of other teleosts and to the mandibular (dentary) barbels of I. punctatus. While the overall histology of the nasal barbel is similar to that of barbels described previously, this study revealed far greater cellular complexity and variability than was previously reported. A layer of stratified epidermal cells rests upon a connective tissue dermis containing a cartilage rod, a large number of nerve fibers and numerous blood vessels, fibroblasts and pigment cells. Taste buds are present in the epidermal layer. This layer was found to contain probably 16 kinds of cell types, several of which may represent transitional stages, in addition to taste bud cells. Observations were made pertaining to innervation and cell types in the taste buds. A new terminology for designating the barbels of I. punctatus is suggested.