Epizootiological study of tick-borne encephalitis virus infection in Japan

Tick-borne encephalitis virus (TBEV) is a zoonotic agent causing severe encephalitis in humans. Rodent species that are potential hosts for TBEV are widely distributed in various regions in Japan. In this study, we carried out large-scale epizootiological surveys in rodents from various areas of Japan. A total of 931 rodent and insectivore sera were collected from field surveys. Rodents seropositive for TBEV were found in Shimane Prefecture in Honshu and in several areas of Hokkaido Prefecture. These results emphasize the need for further epizootiological and epidemiological research of TBEV and preventive measures for emerging tick-borne encephalitis in Japan.     

Immunobiotic lactic acid bacteria beneficially regulate immune response triggered by poly(I:C) in porcine intestinal epithelial cells

ABSTRACT: This study analyzed the functional expression of TLR3 in various gastrointestinal tissues from adult swine and shows that TLR3 is expressed preferentially in intestinal epithelial cells (IEC), CD172a+CD11R1high and CD4+ cells from ileal Peyer's patches. We characterized the inflammatory immune response triggered by TLR3 activation in a clonal porcine intestinal epitheliocyte cell line (PIE cells) and in PIE-immune cell co-cultures, and demonstrated that these systems are valuable tools to study in vitro the immune response triggered by TLR3 on IEC and the interaction between IEC and immune cells. In addition, we selected an immunobiotic lactic acid bacteria strain, Lactobacillus casei MEP221106, able to beneficially regulate the anti-viral immune response triggered by poly(I:C) stimulation in PIE cells. Moreover, we deepened our understanding of the possible mechanisms of immunobiotic action by demonstrating that L. casei MEP221106 modulates the interaction between IEC and immune cells during the generation of a TLR3-mediated immune response.     

Transcriptome analysis of the duodenum in Wistar rats fed a trypsin inhibitor derived from squid viscera

To investigate the effects of oral administration of a trypsin inhibitor (TI), normal Wistar rats were fed a TI derived from squid (Todarodes pacificus) for 10 weeks and gene expression profiles in the duodenum, pancreas, liver, and muscle were then analyzed using DNA microarrays. Although no significant changes could be observed in growth, food intake, tissue weight, or blood tests among the tissues tested, the duodenum showed the most remarkable changes in the global gene expression profile. Significant up-regulation of mRNAs encoding gastrin, gastrokine, cholecystokinin and somatostatin in the duodenum was validated by qPCR analysis. In gene ontology (GO) analysis of the up-regulated differentially expressed genes (DEGs), GO terms related to keratinization and innate mucosal defense were enriched (p < 0.001) in the category of biological processes in addition to assumable terms such as regulation of secretion and response to nutrients, vesicle-mediated transport, and so forth. In the same analysis, calcium ion binding was listed at the deepest hierarchy in the category of molecular function. These results indicate that the duodenum responds to TI treatment by a wider range of physiological processes than previously assumed such as keratinocyte differentiation and innate mucosal defense, in which calcium plays a crucial role.     

Ontogenetic changes of alpha-melanocyte-stimulating hormone (α-MSH)-like immunoreactivity in the brain and hypophysis of a scyliorhinid dogfish (Scyliorhinus torazame): An immunohistochemical study

Ontogenetic development of α-MSH-IR (alpha-melanocyte-stimulating hormone-like immunoreactivity) in the brain and hypophysis of a scyliorhinid dogfish (Scyliorhinus torazame) was studied immunohistochemically. α-MSH-IR first appeared in the hypothalamus of the brain in embryos at the 43-mm stage. Thereafter, α-MSH-positive cells and fibers gradually increased in number in mainly the tuberculum posterius of the hypothalamus during the embyonic periods. In the post-hatching juveniles at the 95-mm and 125-mm stages, α-MSH-positive structures in the brain were comparable to those in adult fish, although the distributional range of α-MSH-IR structures in the brain was less extensive than those in adult fish. In the hypophysis, α-MSH-positive cells first appeared in the caudal part of the adenohypophysis at the 54-mm stage, approximately coincident with the onset of pigmentation in the skin. During ontogenesis, the MSH-producing cells increased in number straightforwardly in the adenohypophysis. This revised version was published online in August 2006 with corrections to the Cover Date.     

Melanocyte-stimulating hormone facilitates hypermelanosis on the non-eyed side of the barfin flounder, a pleuronectiform fish

A pleuronectiform fish, the barfin flounder, Verasper moseri, is promising for aquaculture and resource enhancement in Northern Japan due to its high commercial value. Hypermelanosis of its non-eyed side, which frequently occurs under culture conditions, diminishes its commercial value. Two peptide hormones, melanin-concentrating hormone (MCH) and melanocyte-stimulating hormone (MSH), having opposing actions, are associated with the color changes of fish. We have previously reported the positive effect of MCH in preventing hypermelanosis. Here, we examined the effects of MSH on the occurrence of hypermelanosis. A single injection of Des-Ac--MSH [0.01 nmol/g–10 nmol/g (0.016 μg–16 μg/g)] did not change the eyed-side body color, while a single injection of MCH [0.1 nmol/g (0.21 μg/g)] made the eyed-side color paler. No difference was observed in eyed-side lightness between fish injected with MCH (0.1 nmol/g) and those receiving MCH (0.1 nmol/g) and an increased amount of Des-Ac--MSH (0.01 nmol/g–10 nmol/g) simultaneously. These results indicate that MSH does not suppress the in vivo body color-paling effects of MCH in barfin flounders. On the other hand, implantation of a cholesterol pellet containing Des-Ac--MSH (280 μg, twice at 29-day interval) increased hypermelanosis of the non-eyed side of barfin flounders compared to control fish. Eyed-side bodies of MSH-treated fish were darker than control fish; thus, MSH is involved in morphological color change including ectopic melanin synthesis in non-eyed-side skin.     

Insulin-like growth factor I of Japanese eel, Anguilla japonica: cDNA cloning, tissue distribution, and expression after treatment with growth hormone and seawater acclimation

In an attempt to understand growth regulation in the Japanese eel, Anguilla japonica, we cloned insulin-like growth factor-I (IGF-I) cDNAs and examined their mRNA expression in several tissues. Two eel IGF-I (eIGF-I) cDNAs encoding preprohormones, eIGF-I-Ea1and eIGF-I-Ea2, were cloned from the liver by polymerase chain reaction (PCR). The preproIGF-Is were identical in signal peptide and mature IGF-I, but different in the E domain—eIGF-I-Ea2 mRNA was 36 bp longer than eIGF-I-Ea1 mRNA. Eel IGF-I was 83–94% identical with that of teleosts, 71% identical with that of dogfish, 87% identical with that of bullfrog and chicken, and 83% identical with that of humans. In both males and females the highest eIGF-I-Ea1 mRNA levels were observed in the liver, with detectable levels also found in the gills, heart, stomach, spleen, kidney, intestine, swim-bladder, muscle, and gonads. eIGF-I-Ea1 mRNA levels in the liver were higher in females than in males whereas in the intestine they were lower than in males. eIGF-I-Ea2 mRNA was detected in all the tissues examined and at similar levels in males and females. In this experiment higher eIGF-I-Ea1 mRNA levels were observed in the liver of larger glass eels than in those of smaller fish. eIGF-I-Ea2 mRNA levels were also higher in larger eels, although they were lower than IGF-I-Ea1 mRNA levels. Both eIGF-I mRNA levels in liver were positively correlated with the body size of the␣glass eels. Intraperitoneal injection of recombinant eel GH (reGH), 0.25 μg g⁻¹ body weight, into glass eels resulted in a significant increase in both eIGF-I mRNAs in the liver 1 day after injection compared with control fish, but no elevation was observed 2 days after injection. Incubation of liver slices with reGH at concentrations of 10, 100, and 1,000 ng mL⁻¹ for 24 h resulted in a significant concentration-dependent increase in the levels of both eIGF-I mRNAs. Higher levels of eIGF-I-Ea1 and Ea2 mRNA were observed in the gills ofseawater-reared eels than in those of freshwater-reared fish, but no differenceswere observed in the whole kidney. These results suggest that IGF-I is involved in the regulation of somatic growth and also in adaptation of the Japanese eel to seawater.     

Predation risk management of sea stars (Asterias amurensis and Distolasterias nipon) by adjusting the density and size of seeded scallops (Mizuhopecten yessoensis): an improvement to local mariculture

Optimizing the release density and size of juvenile commercial species for local ecosystems is a critical issue that should be considered when countering predation impacts. To ascertain whether mariculture production of the Japanese scallop (Mizuhopecten yessoensis) could be enhanced by modifying releasing practices, we experimentally investigated the effects of density and size of scallop seeds on their survival in the presence of two sea star species, Asterias amurensis and Distolasterias nipon, with different predation capacities. Under current mariculture practices, the juveniles are briefly exposed to air just before release; therefore, we also examined how air exposure stress increased the predation risk. Scallop survival in the presence of both sea stars increased by?>?20% by increasing scallop size from 30 to 50 mm. Increasing scallop density (range: 5 to 30 scallops m^?2) increased each individual’s survival in the presence of A. amurensis but had no significant effect on predation by D. nipon. Therefore, the release of smaller quantities of large-sized scallops rather than larger quantities of small scallops is recommended to reduce D. nipon predation. In the presence of sea stars, especially by D. nipon, the predation impact on small scallops increased after just a few hours of air exposure, indicating that air exposure of juvenile scallops should be minimized. Our results will mitigate the economic cost of mariculture by facilitating the determination of optimal release conditions for juvenile scallops.

     

Inheritance of resistance to brown leaf spot disease in gentians

Brown leaf spot caused by Mycochaetophora gentianae is a serious disease on gentian in Japan. Previous studies revealed that Gentiana triflora cultivars were susceptible to M. gentianae and that G. scabra cultivars and their interspecific hybrid cultivars were resistant. We subsequently analyzed the mode of inheritance of resistance to M. gentianae in gentians using several populations derived from crosses between G. scabra and G. triflora. Parental G. scabra and G. triflora lines, F₁ and BC₁ plants were inoculated with a conidial suspension and assessed for susceptibility to M. gentianae. All tested F₁ plants were resistant to M. gentianae, indicating that the resistance of G. scabra was inherited dominantly. Segregation ratios in backcross progenies using G. scabra line OK fit the Mendelian ratio of 1:1, showing the involvement of a single dominant locus. These results indicate that the resistance to M. gentianae in G. scabra line OK is controlled by a single dominant allele designated as gentian brown leaf spot resistance (GBLS-1). Finding this allele should facilitate future breeding efforts to develop resistant gentian cultivars.