Isolation and purification of a protective protein antigen of Erysipelothrix rhusiopathiae

The rapid growth and high survival rate of Erysipelothrix rhusiopathiae was determined using a culture of the bacterium in tryptic soy broth supplemented with 0.3% Tris-hydroxymethyl aminomethane and 0.1% Tween 80 (TT-TS broth). High concentrations of 64, 66 and 43 kDa proteins, which are associated with protection against E. rhusiopathiae infection in mice, were obtained by alkaline treatment of whole cells using 0.05-1 N NaOH. The supernatant of alkaline treated cells (alkaline extract; AE) was stable at alkaline or neutral pH. However, aggregates appeared at neutral pH in the absence of sodium dodecyl sulphate (SDS). A high yield of 64, 66 and 43 kDa proteins was obtained from strain Agata (serovar 5). The proteins were eluted from gel bands following SDS-polyacrylamide gel electrophoresis (SDS-PAGE) of the AE from strain Agata and designated P64 and P43. The amounts of P64 and P43 isolated were 0.7 and 0.3 mg/16 g of wet bacteria, respectively. In a mouse protection test, 50% protective doses (PD50) of P64 and P43 were 0.58 and 0.63 microgram, respectively. Upon Western blotting of the AE, both anti-P64 and anti-P43 antibodies reacted with the 64 and 43 kDa proteins. From these results, it is suggested that P64 is the most effective protective antigen and that P43 (43 kDa protein) is a degradation product of P64. Therefore, the 64 kDa structural proteins are associated with the induction of a protective activity against E. rhusiopathiae infection in mice.     

Improvement of the survival in the seven-band grouper Epinephelus septemfasciatus larvae by optimizing aeration and water inlet in the mass-scale rearing tank

The water flow in larval rearing tanks has been indicated to cause mass mortality of the seven-band grouper Epinephelus septemfasciatus larvae. Therefore, a new aerating method was tested in an actual scale intensive rearing tank (8.0 m in diameter, 1.87 m of water depth, 100 m³ of volume), in which an aerator was positioned at the center of the rearing tank surrounding cylindrical drain (1.2 m in diameter) to generate the flow field, and seven larval rearing trials were performed. The survival rate with the former aeration methods were compared, in which several aerators were located in the rearing tank. The survival rate at 10 days after hatching with the new aeration method (61.5±5.1%, n=7) was approximately three times higher than the former methods (21.2±13.7%, n=6). The flow environment of rearing tanks was also examined by quantifying the flow field, and the relationship between the flow field in the rearing tank, behavior of larvae and survival discussed. It was confirmed that the vertical circulating flow was observed in rearing tanks, and determined effectively the survival and the behavior of grouper larvae in patchiness.     

Estimation of flow in a rearing tank of marine fish larvae by simplified numerical computation—a case of two-dimensional flow

Marine fish larvae are fragile against physical stress. However, few studies have been conducted to evaluate the flow field in a rearing tank, which is assumed to provide a high degree of physical stress to marine fish larvae. The flow field in a rearing tank (volume of 1 m³) is generated by aerators, which are commonly used to provide oxygen.

This paper is a report on the estimation of stationary flow in the rearing tank of marine fish larvae. The larvae are seven band grouper larvae of Epinephelus septemfasciatus, which have a very low survival rate immediately after the hatching of eggs. The experiments of rearing of seven band grouper larvae were carried out using rearing tanks with four aeration rates (1000, 200, 50 ml/min, and no aeration). The effects of aeration on the survival and floating death of seven band grouper larvae were examined. The experiments confirmed that the mass mortality of seven band grouper larvae depends on the flow rate in the rearing tank. Aeration at 200 ml/min resulted in the highest survival and growth rates of grouper larvae.

Larvae-rearing experiments provided evidence that the flow rates of the rearing tanks are very important design aspects of rearing tanks. The estimation of flow in a rearing tank for an aerating rate of 200 ml/min was carried out by numerical calculation. The computation was simplified by a two-dimensional flow based on experimental results. The calculated flow in the rearing tank was compared with the experimental one. The calculation of the stationary flow in the rearing tank showed good qualitative and quantitative agreement with the experimental results. The numerical estimation of the flow in a rearing tank of marine fish larvae was confirmed to be effective and satisfactory for the design of a tank that would provide optimum performance.     

Thymus and reproduction: sex-linked dysgenesia of the gonad after neonatal thymectomy in mice

Neonatal thymectomy of mice, when no ectopic thymus existed, constantly resulted in developmental arrest of the ovary but not of the testis; it also caused sterility in the female. The ovaries of thymectomized mice were extremely small and were characterized by absence of follicles and corpora lutea. Such an ovarian dysgenesia was observed when the mice were thymectomized at 3 days of age, but not at 7 days or later; it was prevented by thymus grafting.     

External factors that regulate movement in the marine rotifer Brachionus plicatilis

Fisheries Science - There are two types of movement pattern in the Brachionidae (rotifers), swimming and attachment, although the factors that induce a shift between them have not been adequately...     

Fish‐processing wastes as an alternative diet for culturing the minute rotifer Proales similis de Beauchamp

We evaluated the suitability of fish waste diet (FWD) for culturing the minute rotifer Proales similis through the observation of their population growth and particle size selective feeding. A total of five treatments either with Nannochloropsis oculata or FWD (0.75 and 0.50 g/L) or the combination of N. oculata and FWD were set up in triplicates. P. similis were cultured in diluted natural seawater (8 g/L salinity) and 26 ± 1°C with the diet treatments being applied randomly. We daily monitored the rotifer density, the number of bacteria and the water quality in all the cultures. The population density of P. similis increased exponentially in all treatments, while the mean growth in FWD 0.75 g/L was significantly higher (p < .05) than that in the control. Addition of N. oculata to FWD resulted to lowered P. similis population growth. Bacterial colony count was high in FWD and introduction of P. similis to the diet decreased their density. The estimated bacteria ingestion rates were generally in the range of 6.03 × 10²–1.24 × 10⁴ bacteria/rotifer/hr and there was a positive linear relationship between bacterivory and rotifer population growth. We also observed a shift in the particle size distribution with a reduction in the frequency and concentration of small‐sized particles (<2.5 μm) at day 6. These results accentuate the potential of fish‐processing waste as diet for culturing P. similis which feeds on bacteria and small particles (≤2.5 μm) that are by‐products of degradation of this diet.     

Skeletal development and deformities in cultured larval and juvenile seven-band grouper, Epinephelus septemfasciatus (Thunberg)

The seven‐band grouper, Epinephelus septemfasciatus (Thunberg), is currently recognized as a potential new species for aquaculture in Japan. This study describes normal and abnormal skeletal development of the jaw and vertebrae in cultured larvae and early juveniles of E. septemfasciatus. The ontogenetic stages at which skeletal deformities of jaw and vertebra developed were also described for this species. Osteological observations were made using a clearing and staining method for larvae and soft X‐ray photographs for juveniles. A high incidence of skeletal deformities was observed in the jaws and vertebral column during the larval and juvenile stages. Most of the jaw deformities were explained by an abnormal maxilla curvature. Jaw deformities were visually evident from flexion stage after ossification of the deformed elements. Deformities in the vertebral column (mostly lordosis) were observed from the post‐larval stage and became more evident as growth proceeded. The lordosis generally occurred on the positions of the 8–11th vertebra. These types of deformities are compared with those of other species, and possible causative factors of the skeletal deformities are discussed.
     

Captive Breeding of Threatened African Carp,Labeo victorianus,of Lake Victoria

African carp, Labeo victorianus, is one of the threatened endemic fishes of Lake Victoria which requires conservation and has potential for aquaculture. We conducted a series of experiments on L. victorianus aiming at producing juveniles for both human consumption and stock enhancement. A total of 79 broodstock (mean body weight = 227.06 g) were collected from the wild; packed at 200, 300, and 500 g/L; and transported by road for 7 h. All fish survived during transportation whereas dissolved oxygen decreased and total ammonia nitrogen increased significantly after transport. Females produced 46,702–86,712 eggs (mean = 68,189 ± 2397) and fecundity was highly correlated with the fish body size (R² = 0.96). Only males re‐spawned in captivity while females did not. At 28 C, females spawned 11–12 h after pairing with males. Larvae can either be fed with formulated diet or natural zooplankton. Juveniles (mean length = 36.7 mm; mean body weight = 0.46 g) can be packed as high as 60 fish/L and transported by road for 12 h with low mortality (2.7–10.3%). These results show that captive breeding of L. victorianus can be a good strategy to produce juveniles to boost wild population as well as a source of seeds for culture.