Development of new self-feeding system for mass rearing of ayu <Emphasis Type="Italic">Plecoglossus altivelis altivelis</Emphasis> under artificial and natural light-dark cycles

Self-feeding of grouped ayu Plecoglossus altivelis altivelis was examined under an artificial light-dark (LD) cycle and natural day length using a trigger with a photosensitive sensor. In Experiment 1, approximately 15 juvenile fish (mean body weight, 0.6 g) were kept in 60-L glass tanks (four tanks) under LD 12∶12 (lighting period 06∶00–18∶00 hours) and self-feeding and locomotor activities were simultaneously recorded. Self-feeding was detected within one day and feeding activities were strongly synchronized to the LD cycle, with almost strictly diurnal feeding (98.2%). Although locomotor activities in the upper part of the tank were also greatest during the light phase (76.8%), the activity profiles were different from those of self-feeding, suggesting the validity of the photosensitive sensor. In Experiment 2,25 fish (mean body weight, 15 g) were kept in outdoor 1000-L tanks (four tanks) under natural day length and water temperatures in May, and self-feeding was recorded from mid-August. Stable self-feeding was observed from early September in all the tanks. Nearly all food demands were diurnal, and usually crepuscular. Somatic growth was seen in all tanks. These results demonstrate that the self-feeding system using a photosensitive sensor is applicable for grouped juvenile ayu as small as 0.6 g and adult fish reared under natural conditions.     

Changes in the immunostaining intensities of follicle-stimulating hormone and luteinizing hormone during ovarian maturation in the female Japanese flounder

The role of gonadotropin (GTH) in the reproduction of the Japanese flounder, Paralichthys olivaceus, was studied by assessing the changes in the apparent activity of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in the pituitary gland during gonadal maturation by immunohistochemical analyses. Corresponding changes in plasma levels of testosterone (T), estradiol-17β (E₂), and 17α,20β-dihydroxy-4–pregnen-3-one (DHP) were also studied. Reared fish at the early spawning to termination stages were sampled from May to August and wild fish at the previtellogenic to termination stages were caught at 3- to 4-week intervals between April and September offshore from the northern mainland of Japan by gill nets. The gonadosomatic index of the reared fish decreased from the early spawning stage to the termination stage, while that of the wild fish increased significantly from the previtellogenic stage to the early spawning stage and decreased thereafter. In the reared fish, the immunostaining intensities of FSH and LH were high during the spawning period, accompanied by high plasma levels of T, E₂, and DHP. In the wild fish, the immunostaining intensities of FSH and LH were low during the previtellogenic stage but increased during the maturing and spawning stages. These results indicate that both FSH and LH are likely associated with oocyte maturation in the Japanese flounder.     

Immunological characteristics and response to lipopolysaccharide of mouse lines selectively bred with natural and acquired immunities

Genetic improvement of resistance to infectious diseases is a challenging goal in animal breeding. Infection resistance involves multiple immunological characteristics, including natural and acquired immunity. In the present study, we developed an experimental model based on genetic selection, to improve immunological phenotypes. We selectively established three mouse lines based on phagocytic activity, antibody production and the combination of these two phenotypes. We analyzed the immunological characteristics of these lines using a lipopolysaccharide (LPS), which is one of the main components of Gram‐negative bacteria. An intense immunological reaction was induced in each of the three mouse lines. Severe loss of body weight and liver damage were observed, and a high level of cytokine messenger RNA was detected in the liver tissue. The mouse line established using a combination of the two selection standards showed unique characteristics relative to the mouse lines selected on the basis of a single phenotype. Our results indicate that genetic selection and breeding is effective, even for immunological phenotypes with a relatively low heritability. Thus, it may be possible to improve resistance to infectious diseases by means of genetic selection.     

Is yield enhancement by CO2 enrichment greater in genotypes with a higher capacity for nitrogen fixation?

Yield responsiveness to elevated CO₂ concentration [CO₂] was previously found to be greater when nitrogen (N) was supplied in adequate amounts; however, it remains unclear whether genotypic differences in N₂-fixing capacity affect yield responsiveness in soybean. We tested the hypothesis that yield responsiveness to elevated [CO₂] in soybean is greater in a genotype with a higher capacity for N₂ fixation. We used three near-isogenic genotypes with contrasting nodulation capacities: super-nodulating, normally nodulating and non-nodulating genotypes. Plants were subjected to two levels of [CO₂] (ambient or elevated: ambient + 200 μmol mol⁻¹) and two temperature regimes (low or high: low + ca. 4-5 °C) using temperature gradient chambers. The super-nodulating genotype exhibited a higher N content in leaves, regardless of [CO₂] and temperature. Photosynthetic rates were enhanced by CO₂ enrichment at earlier growth stages, but not at later growth stages, regardless of genotype. This photosynthetic acclimation was reflected in biomass production in all the genotypes examined. Yield responsiveness to elevated [CO₂] was greater in the nodulating genotypes than in the non-nodulating genotype, but the genotypic differences were obscured between the normally nodulating and super-nodulating genotype, thus our hypothesis was not fully verified.     

Effects of feeding time on approaching behavior to food odor in goldfish

ABSTRACT:   Effects of feeding time on approaching behavior to food odor in goldfish Carassius auratus were examined using the Y-maze flow-through system to test whether the behavior differed according to feeding time and experimental time. Fish were fed at 08.00, 12.00 or 16.00 hours once a day or three times (08.00, 12.00 and 16.00 hours) a day for at least 3 weeks under a natural photoperiod, and at 22 ± 2°C, preference reactions to food odor were tested at 08.00, 10.00, 12.00, 14.00 and 16.00 hours. The fish fed once a day showed significant positive approaching behavior only at the time when they were normally fed. The fish fed three times a day showed no significant positive approaching behavior. These results suggest that goldfish can remember when food is available and/or that there exists a mechanism to inhibit approaching behavior when food is not available.     

Actions of glucocorticoid and their regulatory mechanisms in the ovary

Glucocorticoid (G) directly modulates ovarian functions through binding to G receptor. The actions of G are both agonistic and antagonistic depending on the developmental stage of follicles and corpora lutea (CL). During follicular maturation, G suppresses follicular differentiation by downregulating expression of P450 aromatase and luteinizing hormone (LH) receptor in granulosa cells. During ovulation, G protects the ovulatory follicle from inflammatory damage and promotes luteinization, ensuring a smooth transition of the follicle to CL. Throughout life the ovary is exposed to periodic and sporadic waves of G. The Ovary appears to cope with this situation by locally modulating levels of active G. The primary regulatory mechanism consists of two isoforms of 11β‐hydroxysteroid dehydrogenase (11βHSD) that catalyze conversion between active and inactive G. During follicular maturation the levels of active G are suppressed by the dehydrogenase activity of 11βHSD, whereas during the ovulatory process, levels of active G are further increased by the oxo‐reductase activity of 11βHSD. The expression of these enzymes is under the control of gonadotrpins and local regulatory factors such as cytokines, allowing the mechanism to act in coordination with major reproductive events. Thus the G system is an integral part of ovarian physiology, which ensures that the ovary experiences only beneficial effects of G.     

An Endophytic Actinomycete, Streptomyces sp. AOK-30, Isolated from Mountain Laurel and Its Antifungal Activity

To survey endophytic actinomycetes as potential biocontrol agents against fungal diseases of mountain laurel, young plants of mountain laurel (Kalmia latifolia L.) were used as an isolation source. From a total of 73 actinomycetes isolates obtained from leaves, stems and roots of test plants, Streptomyces sp. AOK-30 was selected, because i) it had a broad and intense antimicrobial spectrum against various yeasts and fungal pathogens of Ericaceae, ii) it grew on the multiplication and rooting media for tissue culture for this plant, and iii) the tissue-cultured seedlings that had been treated with this isolate became resistant to Pestalotia disease without any adverse growth effects such as malformation, dwarfing, discolorization and defoliation. This isolate was identified as Streptomyces sp., based on cultural, physiological and morphological characteristics. The present results indicate that isolate AOK-30 is a potential biocontrol agent against diseases of mountain laurel. Received 10 September 2002/ Accepted in revised form 22 November 2002     

Stereochemical basis for the insecticidal activity of carbamoylated and acylated pyrazolines

Methyl 3-(4-chlorophenyl)-1-[N-(4-chlorophenyl)carbamoyl]-4-methyl-2-pyrazoline-4-carboxylate was converted to corresponding (1R)- and (1S)-phenethyl esters via its carboxylic acid and acid chloride at the C-4 atom to separate the diastereomers. Their configurations were confirmed by X-ray analysis. Both isomers of the (1R)methylbenzyl ester were subjected to transesterification with sodium methoxide to obtain enantiomers of the starting methyl ester. Their insecticidal activity was measured against American cockroaches (Periplaneta americana (L.)) by injection and against house flies (Musca domestica L.) by topical application under various synergistic conditions with metabolic inhibitors. The activity values of the four α-methylbenzyl esters and the R-isomer of the starting methyl ester were similar. The S-enantiomer of the methyl ester was about 10 and 100 times more active than the R-isomer against the cockroach and the fly, respectively. Some N-arylacetyl and N-aryloxyacetyl derivatives of the starting N-(4-chlorophenyl)carbamoyl compound gave very low activity. Conformation-energy profiles for some compounds suggested that the conformation of substituents on the N-1 atom in the pyrazoline ring has a specific role for the potential insecticidal effects.     

Evaluation of immunochromatographic test of Shiga toxin 2e in enrichment cultures of swine edema disease clinical samples

To simplify the diagnosis of swine edema disease, overnight culture supernatants of swine clinical samples were assayed using immunochromatographic test strips we developed previously. Small-intestinal contents, mesenteric lymph nodes, and fecal samples were cultured in casamino acid-yeast extract broth overnight, after which supernatants were loaded onto immunochromatographic test strips to determine whether they could detect Shiga toxin 2e (Stx2e). Among 23 clinical samples in which PCR-identified stx2e-positive E. coli were isolated, samples from seven of ten small-intestinal contents, one of three mesenteric lymph nodes and six of ten fecal samples showed Stx2e-positive reactions in the protein-based immunochromatographic test. Additionally, one small-intestinal content sample, in which stx2e-positive E. coli were not isolated, showed an Stx2e-positive reaction. Furthermore, the immunochromatographic test results of the samples were associated with the toxin concentration determined by sandwich ELISA and cytotoxicity assay results on Vero cells. The toxin concentration range of the samples with positive and negative reactions were 2.1–196.2 ng/ml and 0–12.8 ng/ml, respectively. The sensitivity and specificity of this immunochromatographic test strip calculated from all clinical samples analyzed in this study were 60.9% and 94.4%, respectively. Our immunochromatographic test strip has strong potential for simple and accurate diagnosis for edema disease by detecting toxin expression, complementing the PCR method.