Phospholipase A<Subscript>1</Subscript> activity of crude enzyme extracted from the ovaries of skipjack tuna

The phospholipid class composition, fatty acid composition and phospholipase A₁ (PLA₁) activity from the ovaries of skipjack tuna were compared with those of six other species of marine fish. In the skipjack ovaries, the lysophosphatidylcholine (LPC) proportion for the phospholipid, the docosahexaenoic acid (DHA) percentage for the total fatty acids of the phospholipids and the PLA₁ activity of the crude enzyme were the highest among those of the seven species. The optimum pH and temperature for the PLA₁ activity of the crude enzyme from the skipjack ovaries were in the range of pH 6–7 and 20–30°C, respectively, and calcium ions were not required. As a substrate, phosphatidylcholine was more easily hydrolyzed than phosphatidylethanolamine by this enzyme, and the plasmalogen-type phospholipid was much lower than the acyl-type phospholipid. After a 6-h hydrolysis reaction of the purified phospholipid extracted from the mixed ovaries of skipjack and yellowfin tuna by this enzyme, the LPC ratio of the phospholipid increased from 20 to 72.6% and the percentage of DHA for the total fatty acids of the phospholipid also increased. Thus, skipjack ovaries might possibly be used as a source of PLA₁.     

Identification of the feline CD63 homologue using retrovirus-mediated expression cloning

Recently, we combined a retrovirus-mediated expression cloning with a simple screening method using non-adherent cells and panning [Anal. Biochem. 315 (2003) 138]. In this study, we applied this method to identify the antigen recognized by an uncharacterized monoclonal antibody raised against a feline cell line, and identified it as the feline homologue of CD63. This simple method is useful for characterizing unknown antibodies that recognize cell surface molecules. Furthermore, the monoclonal antibody identified as an anti-feline CD63 antibody will be useful for studying feline molecular function(s).     

Histopathological evaluation of the ocular-irritation potential of shampoos,make-up removers and cleansing foams in the bovine corneal opacity and permeability assay

The bovine corneal opacity and permeability (BCOP) assay is an alternative method to the in vivo Draize eye test in rabbits for evaluating eye irritation in vitro. Here, we compared the numerical results of the BCOP assay with the corresponding histopathology for three different corneas for each test substance, including commercially available shampoos, make-up removers and cleansing foams that contained surfactants and other ingredients. The histopathological score was defined based on the severity of lesions in the corneal epithelium. The histopathological findings and scores of the three sections for each test substance were comparable. The in vitro irritancy score (IVIS) generally corresponds to the corneal irritant potential of the test substances assigned on the basis of the histopathological findings in this study. In the present study, we characterized the histopathology of the corneal epithelium and stroma and especially showed that the corneal epithelial injury caused by test substances might be important in assessment of test substances that are mild eye irritants (category 2B) as classified by the United Nations (UN) Globally Harmonized System of Classification and Labelling of Chemicals (GHS), as corneal lesions suggestive of classification into category 2B were localized on the border between the corneal epithelium and stroma, which contained cell elements related to assessment of prognosis of an in vivo eye injury. Histopathological assessment might be useful in predicting in vivo ocular irritation, particularly for test substances with an IVIS >3.1 but ≤25 that are classified as mild irritants (category 2B) according to the UN GHS.     

Motility and fertility of boar semen after liquid preservation at 5°C for more than 2 weeks

This study investigated the effects of skim milk on the quality and fertility of boar spermatozoa under long‐term chilled preservation. Semen samples were stored in Modena solution supplemented with 0 (control) to 50 mg/mL skim milk at 5°C for 4 weeks; spermatozoa stored with 7.5 and 15 mg/mL of skim milk (7.5‐SM and 15‐SM groups, respectively) exhibited significantly higher motility indices than those of the control group up to 3 weeks (P < 0.05), and the 7.5‐SM group showed improved motility indices even after 4 weeks (P < 0.05). In vitro fertilization using spermatozoa in the 7.5‐SM and 15‐SM groups stored at 5°C for 2 weeks showed significantly higher fertilization rates of spermatozoa and the development rates to blastocyst than the control group (P < 0.05), and the 7.5‐SM group showed similar rates of fertilization and blastocyst formation in the fresh non‐stored spermatozoa group. After artificial insemination using spermatozoa stored for 2 weeks in the 7.5‐SM group, healthy piglets were obtained. Boar spermatozoa can be stored at 5°C in a Modena solution containing skim milk. Supplementation of 7.5 mg/mL skim milk improves boar spermatozoa motility and fertility even after liquid preservation at 5°C for 2 weeks.     

Comparison among Japanese isolates of <Emphasis Type="Italic">Pseudomonas savastanoi</Emphasis> pv. <Emphasis Type="Italic">savastanoi</Emphasis>, causal agent of olive knot disease

Olive knot disease in Japan was first reported in Shizuoka Prefecture in 2014, and the causal agent was identified as Pseudomonas savastanoi pv. savastanoi. Subsequently, olive trees having knots were also found in Aichi and Kanagawa Prefectures in 2015, and the isolates from knots were also suspected to be P. savastanoi pv. savastanoi through preliminary examinations. Therefore, the Aichi and Kanagawa isolates were identified through comparison of isolates from three prefectures. Phylogenic analysis based on 16S rDNA and housekeeping genes (gyrB, rpoD, gltA and gap1) revealed that the isolates belonged to the same cluster as the pathotype strain, ICMP4352^PT. The iaaM, H and L genes, which are involved in promotion of symptoms, and the ina gene coding the ice nucleation protein, were detected by PCR from all the isolates. In rep-PCR (ERIC and REP) analyses, the isolates yielded DNA fragment-banding patterns that were nearly identical to that of ICMP4352^PT, but slight variations in banding patterns were observed among them. In a pathogenicity test, the isolates formed distinct knots on olive and pink jasmine. Phenotypic properties of the isolates were almost identical to those of ICMP4352^PT, with the exception of d-sorbitol utilization. Consequently, Aichi and Kanagawa isolates from olive were identified as P. savastanoi pv. savastanoi, and several genetic diversities in terms of rep-PCR were found in the Japanese population of P. savastanoi pv. savastanoi, indicating their heterogeneity.     

Differentiated predation risk on hatchery-reared juvenile masu salmon by white-spotted charr with different body sizes

Predation after release is one of the major concerns of hatchery fish propagation. However, size-specific interaction between predator and prey on the survival of hatchery-released salmonid fish is largely unknown. To understand the size-selective predation risk, 24-h predation experiments were conducted on masu salmon Oncorhynchus masou in tanks. Four ranges of fork length (FL) were examined for masu salmon as a prey, in combination with three ranges of FL for white-spotted charr Salvelinus leucomaenis as a predator. The results show that not only predator and prey sizes, but also interaction between prey size and predator size, strongly affected the survival rate of masu salmon. Predation on masu salmon with the FL exceeding 40% of the FL of white-spotted charr was rare in the experiment. A logistic regression suggests that 37% relative FL of masu salmon to white-spotted charr results in the 50% survival of masu salmon. Our results suggest that adjusting relative size of hatchery fish to the size of local fish predators at the time of hatchery release will have a significant impact on the survival of hatchery fish in the wild. From this perspective, site-specific, adaptive management might be important to improve the effectiveness of hatchery fish propagation.     

Effects of immersion salinity on the food properties of shucked oysters

The effects of immersion salinity on the food properties [water content, salinity, and free amino acid (FAA) content] of shucked oysters were analyzed. Results of a laboratory immersion experiment suggested that the molluscous parts (other than the adductor muscle) swelled in lower salinity and shrank in higher salinity. Higher FAA content was observed in oysters immersed in higher-salinity water. In the adductor muscle, water content increased and FAA content decreased markedly following immersion, regardless of salinity, probably because of intake of immersion fluid and leakage of FAAs across the cut end of the adductor muscle. Immersion salinity ranged from 0.17 to 1.54 % in shucked oyster products on the retail market. Tissue salinity was strongly correlated with immersion salinity (r = 0.904), and tissue water content was correlated negatively with immersion salinity (r = ?0.668). In addition, total FAA and taurine content of oysters were correlated with immersion salinity (r = 0.629 and 0.865, respectively). These results clearly indicate that immersion salinity is an important factor affecting the food components of shucked oysters.     

Recirculation rate of rearing water affects growth of Japanese smelt Hypomesus nipponensis larvae

Larvae of Japanese smelt Hypomesus nipponensis have unique salinity adaptability and a small gape size; consequently, no system has been developed to feed them for stable high growth and survival. Therefore, suitable conditions for larval growth of Japanese smelt remain to be studied. In an earlier study, we developed a Japanese smelt rearing method using a closed recirculating system. Using that system for this study, we examined recirculation rate effects on Japanese smelt larval growth because the recirculation rate can affect larval feeding opportunities through regulation of the rotifer retention time in the rearing tanks. Using 0.2–0.3% salinity and water temperatures of less than 20 °C, we fed Japanese smelt larvae with SS-type rotifer and commercial feed. Results indicated that larval growth with a recirculation rate of less than 648% per day (LR) is superior to that achieved at a rate of over 2160% per day (HR). Moreover, rotifer density under the LR condition declines more slowly than under the HR condition. These results demonstrated that the recirculation rate affects their growth, perhaps by affecting the opportunities for larvae to ingest the rotifers. Furthermore, SS-type rotifers are available for feeding teleosts in a wide range of water temperatures and salinities.