Scatter-spawning of the catfish,Silurus asotus

Reproductive behaviour of the catfish,Silurus asotus was studied in temporary waters around paddy fields. Spawning occurred nocturnally during the first week from the initiation of irrigation. In reproductive activities, a male first energetically pursued a female with its head near to the female’s belly (chasing) and then began to cling to the female’s body from the side, bending its tail or head (clinging). Finally the male enfolded the female’s body, with its anus near to the female’s (enfolding). In some cases, 2–4 males pursued a single female and two males enfolded a female at the same time. Although no aggressive behaviour was evident between males, it was always the largest male that could most frequently approach and enfold the female. The mating pair moved a long distance in a ditch, paddy field and/or creek, performing reproductive activities. It is thought that the spawning site and period of spawning of the fish enable the larvae to avoid the danger of predation and to efficiently feed, firstly on plankton and later on larvae of other fishes which become abundant during the irrigation period. Although some eggs and larvae may die due to the drying out or high water temperatures of such unstable temporary waters, scattering eggs may reduce the incidence of the annihilation of the young.     

Tandem repeat structure of rhamnose-binding lectin from catfish (Silurus asotus) eggs

The primary structure of catfish (Silurus asotus) egg lectin (SAL) was determined. SAL cDNA contained 1448-bp nucleotides and 308 amino acid residues, deduced from open reading frame. The SAL mature protein composed of 285-amino acid residues was followed by a predicted signal sequence having 23 residues. The mRNA of SAL was found to be expressed in eggs, but not in liver. SAL is composed of three tandem repeat domain structures divided into exactly 95 amino acid residues each, and all cysteine positions of each domain were completely conserved. Sequence homologies between the three domains, termed D1 (1-95), D2 (96-190) and D3 (191-285), were as follows; D1-D2, 28%; D2-D3, 33%; D1-D3, 43%. Two conserved peptide motifs, -(AN)YGR(TD)S(T)XCS(TGR)P- and -DPCX(G)T(Y)KY(L)-, appear to exist at the N- and C-terminal regions of each domain, respectively. The kinetic parameters of SAL obtained by measuring surface plasmon resonance were as follows: K(a) (M(-1)) for neohesperidosyl-BSA, 7. 1 x 10(6); for melibiosyl-BSA, 4.9 x 10(6); and for lactosyl-BSA, 5. 2 x 10(5). These results show that RBLs including SAL comprise a family of alpha-galactosyl binding lectins having characteristic tandem repeat domain structures.     

Difference in response by two cyprinid species to predatory threat from the nocturnal catfish Silurus asotus

Response to predators may not be identical between different prey species with different life histories and body sizes, particularly when the threat of predation is not great. To clarify this hypothesis, we introduced two prey species (10 Japanese dace, Tribolodon hakonensis, and 10 pale chub, Zacco platypus) into each experimental pond (in total, 8 ponds×4 trials) in which benthic algae had been allowed to grow. The presence or absence of Far Eastern catfish, Silurus asotus, and a refuge for prey fish was used to produce four treatments. The presence of catfish and/or a refuge did not affect either the feeding behavior or growth rate of Japanese dace. In contrast, when catfish were present and no refuge was available, the incidence of bottom feeding for pale chub greatly decreased. Pale chub growth rate was low when catfish were present and a refuge was available, indicating that pale chub spent more of their time in the refuge and lost opportunities of acquiring food. Japanese dace can reach a threshold size at which the prey are safe from predation, but pale chub cannot, and this may explain the differences in response to predators of the two species.     

Stereotyped sequence of mating behavior in the Far Eastern catfish, Silurus asotus, from Lake Biwa

 Mating behavior of the Far Eastern catfish, Silurus asotus (Siluriformes: Siluridae), was observed in a ricefield system facing the shore of Lake Biwa in mid-May to early June in 1990–1997. A set behavioral sequence similar to those of two other silurid fishes, S. biwaensis and S. lithophilus, both endemic to the Lake Biwa system, was observed: “chasing,”“clinging,”“enfolding” while “squeezing” by the male, and “circling” by the spawning pair. This form of mating behavior is quite different from that of S. asotus reported from the Ooi River system in Kyoto Prefecture, which mainly spawns in running water in ditches. Received: April 10, 2001 / Revised: November 5, 2001 / Accetped: November 20, 2001     

Spawning and season affect lipid content and fatty acid composition of ovary and liver in Japanese catfish (Silurus asotus)

The influences of spawning and season on lipid content, lipid classes, and fatty acid composition were assessed in ovary and liver of wild and cultured Japanese catfish (Silurus asotus). The lipid content (7.3+/-1.6 g/100 g wet wt.) of ovary from wild catfish at spawning was higher than that at post-spawn. However, no influence of spawning on the lipid content of liver was observed. Docosahexaenoic acid [DHA, C22:6(n-3)] in ovary lipids was 12.3+/-0.5% of total fatty acids. The percentage of n-7 monounsaturated fatty acids in triacylglycerol from the ovary and liver in the spawning season was high. Percentages of C22:6(n-3) in phosphatidylcholine and phosphatidylethanolamine from ovary were higher during spawning than after spawning. No significant differences were observed in the lipid content of ovary and liver from cultured catfish between seasons (summer vs. winter). Content of arachidonic acid (C20:4n-6) in ovary and liver from cultured catfish was higher in summer than in winter. There were differences in lipid classes of ovary and liver by spawning and season. These results suggest that the lipid metabolism in Japanese catfish is greatly influenced by spawning and season.     

Thresholds of retinal and extraretinal photoreceptors measured by photobehavioral response in catfish,Silurus asotus

Summary Light thresholds of retinal and extraretinal photoreceptors in catfish were examined by the photobehavioral response using a method in which reflex body movements are recorded. Thresholds were determined in six groups, (A) intact, (B) ophthalmectomized, (C) pinealectomized, (D) ophthalmectomized + pinealectomized, (E) ophthalmectomized, pinealectomized and skinless over the brain (skinless fish) and (F) ophthalmectomized, pinealectomized and dorsally covered with aluminum foil over the brain (covered fish). All these fishes displayed short term activity to white light stimulation after being dark adapted for more than 5 h. The lowest threshold was obtained in the intact group (2.0×10^–4 W/cm²). The thresholds of lateral eyes and the pineal organ were 3.4×10^–3 and 1.5×10^–2 W/cm², respectively. Without lateral eyes and pineal organ, catfish still responded to light, indicating the possible existence of extraretinal nonpineal photoreceptors (ENPs). The threshold of ENPs was 3.3 W/cm². The localization of ENPs was assumed to be in the brain from the experiment with the combination of skinless and covered fish.Abbreviation ENP extraretinal nonpineal photoreceptor     

Effect of meal size on postprandial metabolic response in Chinese catfish (Silurus asotus Linnaeus)

The effect of relative meal size (0.5–24% body mass) on specific dynamic action (SDA) was assessed in Chinese catfish (Silurus asotus Linnaeus) (30.90±1.30 g) at 25.0°C; the cutlets of freshly killed loach without viscera, head and tail were used as a test meal. There was no significant difference in either SDA duration or peak oxygen consumption (VO₂) among low meal size ranges. But both increased linearly as meal size increased from 2 to 24% without reaching a plateau. Factorial metabolic scope was 5.92 in fish fed with 24% body mass, the highest documented feeding metabolic scope value in fish till now. The Peak VO₂ of satiated meal size groups (175.85±10.55 mg O₂ h⁻¹) was above 80% of maximum metabolic rate during locomotion recovery process (215.48±7.07 mg O₂ h⁻¹). The relationship between energy expended on SDA (E) and energy ingested (I) was described as: E=0.0000432I ²+0.140I+2.12. The lowest value of SDA coefficient appeared at 2% body mass group.     

A unique epidermal mucus lectin identified from catfish (Silurus asotus): first evidence of intelectin in fish skin slime

The present study reports a new type of skin mucus lectin found in catfish Silurus asotus. The lectin exhibited calcium-dependent mannose-binding activity. When mannose eluate from chromatography with mannose-conjugated agarose was analysed by SDS-PAGE, the lectin appeared as a single 35-kDa band. Gel filtration showed that the lectin forms monomers and dimers. A 1216-bp cDNA sequence obtained by RACE-PCR from the skin encoded a 308 amino acid secretory protein with homology to mammalian and fish intelectins. RT-PCR demonstrated that the lectin gene was expressed in the gill, kidney and skin. Subsequent sequencing revealed the presence of an isoform in the gills. Antiserum detected the intelectin protein in club cells in the skin and gill, renal tubules and blood plasma. Although intelectin gene expression was not induced by in vivo bacterial stimulation, the intelectin showed agglutination activity against the pathogenic bacterium Aeromonas salmonicida, suggesting that the lectin plays an important role in self-defence against bacteria in the skin surface of the catfish. These findings represent one of the few examples of characterization and functional analysis of a fish intelectin protein.     

Dietary and seasonal effects on the dorsal meat lipid composition of Japanese (Silurus asotus) and Thai catfish (Clarias macrocephalus and hybrid Clarias macrocephalus and Clarias galipinus)

The effects of dietary lipids and seasonal variation on the lipids of wild and cultured catfish (Japanese catfish, Silurus asotus; Thai catfish, Clarias macrocephalus and hybrid Clarias macrocephalus x Clarias galipinus) were determined by analysis of the lipid content and fatty acid composition of their dorsal meat. The predominant fatty acids of dorsal meat were 16:0, 18:1n-9, 18:2n-6, 20:4n-6 (arachidonic acid, AA), and 22:6n-3 (docosahexaenoic acid, DHA). The DHA content in the diet of Japanese catfish was higher than that in the diet of Thai catfish, and this was reflected in the dorsal meat of the Japanese catfish, which had a remarkably high percentage of DHA compared with the meat of the Thai catfish. Cultured Japanese catfish had a higher percentage of 18:2n-6 than Thai fish and a lower percentage of AA in winter than in summer season. There were also seasonal variations in the percentage of n-6 fatty acids in Japanese catfish. In summer, the fatty acid composition of the cultured Japanese catfish was similar to that of the wild catfish. These fatty acid changes in the lipid classes, triacylglycerol, phosphatidylcholine and phosphatidylethanolamine were similar to those observed for total lipids. These results indicate that the percentage of DHA in the dorsal meat of catfish is influenced by dietary fatty acid, and it may be that it can be increased in cultivated fish by administering a diet containing a large amount of DHA.     

Separation and analysis of the major forms of plasma fibronectin

Human plasma fibronectin exists in circulation in multiple molecular forms that are distinguishable by SDS-polyacrylamide gel electrophoresis (zone I, approx. 450 kDa dimers; zone II, 190-235 kDa; Zone III, 146-175 kDa). (Chen, A.B., Amrani, D.L. and Mosesson, M.W. (1977) Biochim. Biophys. Acta 493, 310-322). We report here on investigations of plasma fibronectin that had been purified from the 'heparin-precipitable fraction' of plasma by DEAE-cellulose chromatography using buffers containing a chaotropic salt (KSCN). Zone I fibronectin and zone II fibronectin were subsequently separated by Sepharose CL-6B chromatography in the presence of 0.3 M KSCN. Electrophoresis of reduced zone I fibronectin dimers showed the presence of three types of subunits (i.e., 220 kDa, 215 kDa, 207 kDa), evidently all having the same NH2-terminal sequence. Subunits of this size were also found in reduced zone II fibronectin, as well as another polypeptide of 190 kDa, the latter amounting to under 5% of the total. Unreduced zone I fibronectin was resolved by gel electrophoresis into a doublet. The upper component amounted to approx. 90% of the total and was comprised of 220 kDa and/or 215 kDa subunits; the lower component contained 207 kDa plus a 220 kDa or 215 kDa subunit. Scanning transmission electron microscopy indicated that under physiologic conditions zone II fibronectin molecules, like those in zone I, exist as pleiomorphic, loosely folded structures (approx. 16 X 8-12 nm) that are somewhat smaller than dimeric zone I molecules (approx. 24 X 16 nm). Circular dichroic spectral analyses suggests that both types have similarly folded local domains. Affinity chromatography experiments revealed a relative decrease in the binding of zone II fibronectin to gelatin but no difference from zone I fibronectin with respect to heparin or fibrin binding.     

Isolation and characterization of rat plasma fibronectin.

Rat plasma fibronectin has been isolated and characterized and monospecific antibodies were prepared to it. Two components of fresh rat plasma (in the presence of proteinase inhibitors) bound to a gelatin-Sepharose affinity column. One protein was eluted with 4.0 M-urea and was identified as fibronectin. Another protein was eluted from the gelatin-Sepharose column with 8.0 M-urea and was identified as a 70 000-Mr collagen-binding molecule. This 70 000-Mr fragment was found to be a normal constituent of blood plasma, and its presence did not represent a proteolytic degradation product formed during isolation. The antibodies prepared against rat fibronectin only weakly cross-reacted with plasma fibronectins of chicken, horse and human. These studies shed light on the metabolic interrelationships between fibronectin and other collagen-binding molecules.     

Isolation and identification of androstanediol glucuronide from human plasma

[3H]Dihydrotestosterone (50 microCi) was infused into normal men and women for 8 h. It was previously shown that this was sufficient time for this material to reach a steady state. Venous plasma was obtained at 6 and 8 h, pooled, and the unconjugated steroids removed by ether extraction. The remaining plasma was adjusted to pH 4.9 and the steroid conjugate was extracted first with ethyl acetate and then with an ether-ethanol mixture. The extracts were combined and taken to dryness. Steroid sulfates were solvolyzed using dioxane, and the mixture partitioned between ether and 1% NaOH. The aqueous phase was acidified and added to an XAD-2 column, washed with water, and the glucuronide fraction eluted with methanol. The solvent was concentrated and the methanol extract was passed through a C18 Sep-Pak, filtered through an Acrodisc CR and then subjected to gradient high performance liquid chromatography [HPLC] (Nova-Pak C18, KH2PO4, pH 3, and methanol). The fractions containing steroid glucuronides were collected and esterified with diazomethane and then acetylated with acetic anhydride in pyridine. The glucuronide triacetyl methyl ester (GAME) derivatives were then run in a second HPLC system (3 Lichrosorb 5 mu columns, 4 mm x 25 cm) using a gradient of ethanol-heptane and heptane. We clearly established that this system separates 3 alpha-diol GAME conjugated at the 17 and 3 positions (44 vs 50 min) with authentic samples previously synthesized in our laboratory. We concluded that the pooled plasma contained only the 17-GAME conjugate. No significant activity of the 3-glucuronide was detected. The natural compound in circulation, therefore, is 5 alpha-androstane-3 alpha, 17 beta-diol 17-glucuronide.     

Resistance of the European catfish (Silurus glanis) to channel catfish virus

European catfish (Silurus glanis) fingerlings (2 to 4 g each) were tested for susceptibility to channel catfish virus (CCV). They had supported CCV replication at 2 days after intraperitoneal injection with 0.1 ml of saline containing 10⁵ TCID₅₀. Homogenized visceral organs (liver, kidney and spleen) contained 10⁴ TCID₅₀/0.1 ml at 2 days post inoculation (PI) but at 4 days the titer decreased to 10¹ TCID₅₀. Bathing European catfish in CCV yielded only one positive sample with à titer of 10^0.83 TCID₅₀ per 0.1 ml of tissue. No clinical signs of CCV developed and no virus related deaths occurred.     

Mating behavior of the rock catfish, Silurus lithophilus

The mating behavior of the rock catfish Silurus lithophilus (Siluriformes: Siluridae), a species endemic to the Lake Biwa system, was observed from May to July in 1989–1994 along the rocky shore of the lake's outlet, the Seta River. The mating behavior of S. lithophilus involved a certain behavioral sequence: “chasing,”“clinging,” and “enfolding” while “squeezing” by the male; and “circling” by the spawned pair. The mating behavior of this species was basically similar to that of S. biwaensis, but greatly different from that of S. asotus, which spawns in running water (in ditches). The mating behavior of S. lithophilus (and S. biwaensis) might have developed as an adaptation to lentic environments such as the shores of the large river or the lake. Received: October 25, 2000 / Revised: February 25, 2001 / Accepted: March 8, 2001     

Isolation and identification of the major concanavalin A binding glycoproteins from murine-lymphocytes.

The major glycoproteins which bind concanavalin A have been isolated and identified from murine spleen cells, thymocytes,and purified thymus-derived (T) lymphocytes, and from the spleen cells of congenitally athymic (nude) mice. The cells were radiolabeled by lactoperoxidase catalyzed 125I iodination or by culturing the cells in media containing [3H]leucine or [3H]fucose. The cell membrane was solubilized with Nonidet P-40 and the concanavalin A binding proteins were isolated by affinity chromatography and analyzed according to their mobility on polyacrylamide gel electrophoresis in sodium dodecyl sulfate. The major proteins from various lymphocyte preparations were identified by immunoprecipitation with specific antisera. The molecules coded by the histocompatibility-2 complex acted as concanavalin A binding proteins H-2K and H-2D were isolated from T lymphocytes, thymocytes, and bone marrow derived (B) lymphocytes. The Ia antigens were identified from B lymphocytes and tentatively identified from T lymphocytes. In addition to these H-2 complex proteins, immunoglobulin M and D on B lymphocytes also bound concanavalin A binding. All these glycoproteins have previously been identified as cell surface molecules. The presence of certain minor unidentified concanavalin A binding proteins on lymphoid cells is indicated.     

Specificity of the collagenolytic serine proteinase from the pancreas of the catfish (Parasilurus asotus)

The collagenolytic serine proteinase from the pancreas of the catfish (Parasilus asotus) had a pH optimum of 7.5 for native, reconstituted calf skin collagen fibrils. The enzyme was most stable at pH 6-9. The enzyme hydrolyzed heat-denatured collagen (gelatin), casein, hemoglobin and elastin in addition to native collagen but not virtually Tos-Arg-OEe, Bz-Tyr-OEe and Suc-(Ala)3-NA. The enzyme cleaved Leu-Gly (or Gln-Gly), Gly-Ile and Ile-Ala bonds on DNP-Pro-Leu-Gly-Ile-Ala-Gly-Arg-NH2 and DNP-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg.     

Isolation and characterization of the major form of beta-glucuronidase from human seminal plasma

Human seminal plasma contain two forms of beta-glucuronidase (beta-D-glucuronidase glucuronosohydrolase, EC 3.2.1.31) which are present in the ratio of 4:1. The major form of beta-glucuronidase with a slow moving band in electrophoresis was purified to homogeneity as revealed by polyacrylamide gel electrophoresis, double immunodiffusion and immunoelectrophoresis. The major form of beta-glucuronidase shows dual optimum pH at 4.3 and 4.7 with a dip in the activity at pH 4.5. The Km of this form of beta-glucuronidase is dependent on pH and was found to be 0.95, 3.08 and 0.67 mM at pH 4.4, 4.5 and 4.7, respectively. The major form of beta-glucuronidase from seminal plasma is stable at 55 degrees C for 30 min but it denatures at 65 degrees C. Heat denaturation is faster at acidic pH (4.7) than at alkaline pH (7.8). However, the activity of enzyme increased linearly with increase in temperature up to 70 degrees C during incubation with substrate. Cu, Ag, Hg and Ni salts inhibited enzyme activity significantly at 0.1 and 1.0 mM concentration, but the inhibition of HgCl2 was protected by cysteine. 1,4-D-Saccharic acid lactone and ascorbic acid inhibited seminal beta-glucuronidase competitively, yielding Ki values of 1.7 . 10(-3) mM and 10.3 mM, respectively. Though fructose and mannose also showed significant inhibition of beta-glucuronidase at 10-100 mM, glucose did not show any effect. The molecular weight of the major form of beta-glucuronidase was found to be 279 000, and it appears to be composed of four subunits each having a molecular weight of 74 000.     

Reproductive ecology of the Far Eastern catfish, Silurus asotus (Siluridae), with a comparison to its two congeners in Lake Biwa, Japan

Spawning activity of the catfish, Silurus asotus, takes place in temporary water (rice fields) from early evening to midnight, generally in connection with rainfall, from late April to late August. Spawning of the catfish was correlated with hydrographic parameters dependent on rainfall: daily precipitation, turbidity, water depth, and water temperature. The spawning habits of the catfish, and in particular the use of temporary waters such as rice fields that become submerged after rainfall or by irrigation, are presumed to be adaptations to the Asian monsoon climate with a pronounced rainy season. The apparent sex ratio of the catfish was extremely biased toward females. Intraspecific variation in the reproductive ecology, particularly mating behavior, of this species is observed among local populations. Factors that may have caused this variation are discussed in the context of a comparison of mating behavior, reproductive environment, and sex ratio between the Lake Biwa population of S. asotus and other conspecific populations, as well as two other species of silurid catfish that occur in the Lake Biwa drainage, S. biwaensis and S. lithophilus.