Repeatable immersion infection with <Emphasis Type="Italic">Photobacterium damselae</Emphasis> subsp. <Emphasis Type="Italic">piscicida</Emphasis> reproducing clinical signs and moderate mortality

Pseudotuberculosis is a bacterial septicaemia caused by Photobacterium damselae subsp. piscicida in several marine fish species. Yellowtail Seriola quinqueradiata is the most sensitive fish species to this disease. The internal organs of naturally infected yellowtail exhibit whitish spots, tubercle-like tissue structures, consisting of bacterial accumulations. There have been many trials for experimental infection, however adequate method of infection that reproduces moderate mortality and primary clinical signs has not yet established. Present investigation evaluated an immersion infection method by using logarithmic culture-phase bacteria resulting in higher mortality than that using stationary culture-phase bacteria. Typical white spots on the spleen and kidney were also observed constantly in dead fish. Transmission electron microscopy and fluorescent antibody microscopy showed bacterial clusters not only in the spleen and kidney but also in the blood channels in the secondary gill filaments. These results were confirmed repeatedly by plural experiments. The use of logarithmic-phase bacteria in immersion infection is an appropriate technique to reproduce moderate mortality and primary clinical signs, which will be a reliable infection method also for the challenge test of pseudotuberculosis vaccine.     

Three cone opsin genes and cone cell arrangement in retina of juvenile Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis>

ABSTRACT:   In bluefin tuna aquaculture, collision of juveniles with the tank or net walls is a major cause of high mortality. This problem may be related to color sensibility of the visual mechanisms of this species. As a first step in understanding of color vision of Pacific bluefin tuna Thunnus orientalis , we applied a molecular technique and histology to study cone cell distribution in the retina of juvenile fish. We isolated three cone opsin genes encoding one blue-sensitive ( SWS2 ) and two green-sensitive ( RH2 ) visual pigments. In situ hybridization revealed that SWS2 mRNA is localized in the single-cone photoreceptors. The localization of the two RH2 signals in distinct cone cells was not determined, probably because of the high homology between the two RH2 genes. Single-cone photoreceptors appeared frequently in the ventral–temporal retina in approximately 80-mm fish and in the temporal retina in approximately 230-mm fish. These cone distributions may define a visual field with best color contrast vision in front and above the fish with a short wavelength (blue) reflecting target (sensed by single cones), and may be enhanced against the longer wavelength (green) background when fish see a target below them (sensed by double cones).     

Prebiotic effect of glycerol galactoside isolated from color-faded nori in rats

Glycerol galactoside (GG; floridoside: 2-O-glycerol-α-d-galactopyranoside, and isofloridoside: 1-O-glycerol-α-d-galactopyranoside) is known to be a component of edible red seaweed nori. Recently, we have found that low-quality nori (color-faded nori), which has a low protein content, contains a large quantity of GG. From further studies, we have also found that GG has prebiotic characteristics in vitro. In this study, we evaluated the in vivo prebiotic activity of GG in rats. Dietary GG selectively increased the cecal Bifidobacterium count in rats. Other indices of prebiotics, such as pH of cecal content, organic acid concentrations, and fecal weight, also supported the existence of prebiotic activity of GG. The present data will also contribute to the development of a new method of utilizing color-faded nori as a health-promoting foodstuff.     

Upgrading of urea formaldehyde-bonded reed and wheat straw particleboards using silane coupling agents

Reed and wheat straw particleboards bonded with urea formaldehyde (UF) resin were manufactured from two different material configurations (i.e., fine and coarse particles). The board densities were in the range of 0.550–.90g/cm³. The effects of particle size and board density on the board properties were examined. The properties of particleboard produced from fine particles were better than those made from coarse particles. An increase in board density resulted in a corresponding improvement in the board properties. The properties of OF bonded reed and wheat straw particleboards were relatively lower than those of commercial particleboards. Three silane coupling agents were used to improve the bondability between the reed and wheat particles and OF resin. Results of this study indicate that all the board properties were improved by the addition of silane coupling agent. The degree of improvement achieved from each coupling agent was different; epoxide silane was found to be more effective for reed straw particleboard, and amino silane was better for wheat straw particleboard.     

Effect of dietary medicinal herbs on lipid metabolism and stress recovery in red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

The effect of dietary medicinal herbs on lipid metabolism and stress recovery was investigated in red sea bream Pagrus major. Fish (mean body weight 24.0 ± 0.2 g) were fed on test fish meal diets supplemented with either Massa Medicata (Mm), Crataegi Fructus (Cf), Artemisia capillaries (Ac), or Cnidium officinale (Co), or with a mixture of the four herbs (HM) for 12 weeks. A control group was fed a diet without herbs in the same manner. A high survival rate was observed in the herbal diet groups. The final mean body weight, feed efficiency, protein efficiency ratio, and apparent protein and lipid retention in the Ac, Co, and HM groups were higher than those in the control and Mm groups. Final carcass, hepatic lipid and triglyceride contents, and plasma triglyceride and nonesterified fatty acid levels were lower in the Ac, Co, and HM groups compared to those of control and Mm groups. However, final hepatic phospholipid, plasma phospholipid, and high density lipoprotein cholesterol levels were higher in the Ac, Co, and HM diets groups than in the control and Mm groups. The Cf, Co, Ac, and HM groups showed faster recovery time in the 2-phenoxyethanol anesthesia test and a higher recovery rate in the 10-min air exposure test than those of the control and Mm groups. Moreover, the Cf, Ac, Co, and HM diet groups had a significantly lower plasma cortisol level than the control and Mm diet group, but the glucose level in the herbal diet groups was higher than that in the control group after a 1-h air exposure. These results indicate that the addition of medicinal herbs to the fish diet improved lipid utilization and stress recovery in red sea bream.     

Role of the rice blast resistance gene Pi-cd in rice (Oryza sativa) breeding programmes

A series of DNA markers for various agronomic traits may accelerate the success of marker-assisted selection in practical plant breeding programmes. Here, we developed DNA markers for the blast resistance gene Pi-cd. In this study, we examined the effects of the Pi-cd locus on not only blast resistance but also agronomic traits in agriculture. We developed three pyramiding lines (PLs) coupling Pi-cd with three blast resistance genes, pi21, Pi35 and Pi39. The effect of Pi-cd on blast resistance was dependent on the coupled resistance genes. Then, we evaluated the effects of Pi-cd on 13 agronomic traits. Amylose content and 1,000-grain weight showed significant differences between the PLs and current commercial varieties, which had no negative effects on agronomic trait values. Furthermore, we investigated the distribution of genotype for the Pi-cd locus among varieties of upland rice. The KT genotype specific to rice blast resistance may be predominant in the varieties. The results suggested that Pi-cd has the potential to be useful for improving blast resistance in rice breeding programmes.     

Stimulation of growth performance without causing stress response in young red sea bream, Pagrus major (Temminck & Schlegel), by photoperiod manipulation

Three photoperiods (12L:12D, 16L:8D and 24L:0D) were used to investigate the growth performance and stress response in red sea bream, Pagrus major (body weight 200–400 g). Fish were fed a commercial diet to apparent satiation, two times a day for 8 weeks. Fish exposed to a 24L:0D photoperiod showed a significantly higher weight gain (%) than those exposed to other photoperiods (P<0.05). The best specific growth rate and feed conversion efficiency were achieved at 24L:0D and 16L:8D, without significant differences among them. Although fish exposed to 16L:8D showed a significantly higher plasma level of cortisol than those exposed to other photoperiods in the third week, there was no major variation in the cortisol level among the treatments either at the sixth week or at the end of this study. There were no significant differences either in the haematocrit level or the plasma levels of glucose, total cholesterol and total protein among the treatments during this study. The results revealed that the growth performance of red sea bream reared from 200 to 400 g can be stimulated significantly using a continuous (24L:0D) photoperiod without any measurable significant stress response in fish.     

Expression of O6-methylguanine-DNA methyltransferase causes lomustine resistance in canine lymphoma cells

The DNA repair protein O⁶-methylguanine-DNA methyltransferase (MGMT) causes resistance to nitrosoureas in various human cancers. In this study, we analyzed the correlation between canine lymphomas and MGMT in vitro. Two of five canine lymphoma cell lines required higher concentrations of lomustine to inhibit cell growth by 50%, but their sensitivity to the drug increased when they were cultured with an MGMT inhibitor. Fluorometric oligonucleotide assay and real-time polymerase chain reaction of these cell lines revealed MGMT activity and high MGMT mRNA expression, respectively. We analyzed the methylation status of the CpG islands of the canine MGMT gene by the bisulfite-sequencing method. Unlike human cells, the canine lymphoma cell lines did not show significant correlation between methylation status and MGMT suppression levels. Our results suggest that in canine lymphoma MGMT activity may influence sensitivity to nitrosoureas; thus, inhibition of MGMT activity would benefit nitrosourea-resistant patients. Additional studies are necessary to elucidate the mechanism of regulation of MGMT expression.     

The role of neighboring infected cattle in bovine leukemia virus transmission risk

A cohort study was conducted to evaluate the risk of bovine leukemia virus (BLV) transmission to uninfected cattle by adjacent infected cattle in 6 dairy farms. Animals were initially tested in 2010–2011 using a commercial ELISA kit. Uninfected cattle were repeatedly tested every 4 to 6 months until fall of 2012. The Cox proportional hazard model with frailty showed that uninfected cattle neighboring to infected cattle (n=53) had a significant higher risk of seroconversion than those without any infected neighbors (n=81) (hazard ratio: 12.4, P=0.001), implying that neighboring infected cattle were a significant risk factor for BLV transmission. This finding provides scientific support for animal health authorities and farmers to segregate infected cattle on farms to prevent spread of BLV.     

Anatomical evaluation of hepatic vascular system in healthy beagles using X-ray contrast computed tomography

Liver contrast X-ray computed tomography (CT) has been used for evaluation of hepatic vessels for liver transplantation, liver lobectomy, interventional radiology and diagnosis of hepatocellular carcinoma in humans. However, there remains scant available anatomical information on normal hepatic vessels in the veterinary field. In this study, visualization of hepatic vessels was evaluated in 32 normal beagle dogs by X-ray contrast CT using triple phase images. The following hepatic vessels were clearly visualized: arterial, portal and hepatic veins. With regards to the running patterns of the portal vein and hepatic vein, there were no significant differences between the dogs. However, the hepatic artery exhibited some differences in each dog. In particular, the hepatic artery of the quadrate lobe and the right lateral lobe had many running patterns. The results of the present study could be useful for veterinary diagnosis, surgery and interventional radiology.