Suitability of genetically modified soybean meal in a dietary ingredient for common carp Cyprinus carpio

The effect of genetically modified (GM) soybean meal (SBM) in a feed ingredient on growth performance of common carp was investigated in comparison to nonGM SBM. GM SBM was included at 34 and 48% in two experimental diets that were formulated with fish meal (FM) to obtain approximately 38% protein in diet. Two other experimental diets were formulated to contain the same levels of nonGM SBM. The diets were fed to juvenile common carp (22 g initial mean weight) for 12 weeks. The cauliflower mosaic virus (CaMV) 35S promoter fragment (205 base pairs) of the GM SBM was examined in fish muscle and blood samples at the twelfth week. From the twelfth week, the GM groups were fed with nonGM diets to determine the residual span of the transferred promoter fragment. There was no significant difference in growth and feed performance between GM and nonGM groups at two inclusion levels after 12 weeks. The CaMV 35S promoter fragment was not detected in fish muscles or blood receiving either level of GM SBM diet. The results demonstrated that the availability of GM SBM was similar to that of nonGM SBM and the GM SBM would be a suitable and safe ingredient in feed for common carp.     

Assessment of decay risk of airborne wood-decay fungi

The decay risk of airborne wood-decay fungi was investigated by using an air sampler. Japanese cedar disks 7.8 cm in diameter and about 3 mm in thickness with moisture content of about 100% were placed in a “BIOSAMP” air sampler and exposed to 1000 l air. Air sampling was carried out from June to September at the same sampling site in Tsukuba, Japan. The exposed disks were then incubated for 16 weeks in a damp container kept at 26° ± 2°C. During the incubation period, wood mass loss ranged from −15 to 807 mg with a mean mass loss of 244 mg. Factors affecting mass loss were explored. Wood moisture content and ratio of heartwood area proved to be significant factors. In addition, six weather factors were found to influence mass loss. Disks that were sampled on a cloudy day showed significantly higher mean mass loss compared to those sampled on a sunny day. Subculturing of filamentous fungi from 16-week incubated disks suggested one-third of the isolated fungi produced ligninolytic enzymes.     

Molecular characterization and expression analysis of heat shock proteins 40, 70 and 90 from kuruma shrimp <Emphasis Type="Italic">Marsupenaeus japonicus</Emphasis>

Heat shock proteins (HSPs) are proteins that are expressed more strongly when the cells are exposed to physiological and stressful conditions. In this study, the full-length cDNAs of heat shock proteins 40 (MjHSP40), 70 (MjHSP70) and 90 (MjHSP90) were cloned from kuruma shrimp Marsupenaeus japonicus. The open reading frames (ORFs) of the cDNA clones have lengths of 1,191, 1,959 and 2,172 bp and encode 396, 652 and 723 amino acid residues, respectively. The predicted MjHSP40 amino acid sequence contains a J domain, a glycine/phenylalanine-rich region, and a central domain containing four repeats of a CxxCxGxG motif, indicating that it is a type I HSP40 homolog. The signature sequences of the HSP70 and HSP90 gene families are conserved in the MjHSP70 and MjHSP90 amino acid sequences. The deduced amino acid sequences of MjHSP70 and MjHSP90 share high identity with previously reported shrimp HSP70s and HSP90s, respectively. The expression of MjHSP90 mRNA increased at 32°C. Additionally, the expressions of MjHSP40, MjHSP70 and MjHSP90 mRNAs increased in defense-related tissues (i.e., hemocytes and lymphoid organ) when the shrimp were challenged with white spot syndrome virus.     

The existence of aspolin and its trimethylamine-<Emphasis Type="Italic">N</Emphasis>-oxide demethylating activity in the muscle of freshwater fish

ABSTRACT:   Aspolin is a polyaspartic acid-like protein, which is originally isolated from walleye pollack Theragra chalcogramma muscle as trimethylamine- N -oxide (TMAO) demethylase. Although carp Cyprinus carpio muscle contains a trace amount of the enzyme substrate, TMAO, aspolin can be extracted and purified by acid treatment, successive chromatographies and polyacrylamide gel electrophoresis, and has twice the amount of that in walleye pollack muscle. Carp aspolin showed a low enzymatic activity in the presence of Fe²⁺ and reductants, and its Km value (100 mM) to TMAO was extremely high. It was a thermostable protein and had an unfolded conformation. The amino acid sequence of carp aspolin 1 deduced from cDNA revealed that it contained a long Asp polymer, an uninterrupted stretch of 138 Asp residues, followed by four amino acid residues, His-Glu-Glu-Leu, in C-terminus. The chain length was shorter by 42 Asp residues than that of its walleye pollack counterpart.     

Anti‐PirA‐like toxin immunoglobulin (IgY) in feeds passively immunizes shrimp against acute hepatopancreatic necrosis disease

Acute hepatopancreatic necrosis disease (AHPND), caused by a toxin‐producing Vibrio parahaemolyticus strain, has become a serious threat to shrimp aquaculture. The need to regulate antibiotic use prompted the development of alternative ways to treat infections in aquaculture including the use of chicken egg yolk immunoglobulin (IgY) for passive immunization. This study evaluated the protective effect of IgY against AHPND infection in Litopenaeus vannamei (Boone). IgY was isolated from eggs laid by hens immunized with recombinant PirA‐like (rPirA) and PirB‐like (rPirB) toxins. Whole‐egg powders having IgY specific to rPirA (anti‐PirA‐IgY) and rPirB (anti‐PirB‐IgY) and IgY from non‐immunized hen (control‐IgY) were mixed with basal diets at 20% concentrations and used to prefeed shrimp 3 days before the bacterial challenge test. Survival rates of the challenged shrimp fed the anti‐PirA‐IgY, anti‐PirB‐IgY and control‐IgY diets were 86%, 14% and 0%, respectively. Only the feed containing anti‐PirA‐IgY protected shrimp against AHPND. Increasing the concentration of rPirA antigen to immunize hens and lowering the amount of egg powder in feeds to 10% consistently showed higher survival rates in shrimp fed with anti‐PirA‐IgY (87%) compared with the control (12%). These results confirm that addition of anti‐PirA‐IgY in feeds could be an effective prophylactic method against AHPND infection in shrimp.     

Effects of purple sweet potato anthocyanins on development and intracellular redox status of bovine preimplantation embryos exposed to heat shock

The development of cleavage stage preimplantation embryos is disrupted by exposure to heat shock, such as high temperatures in the summer season. In this study, we investigated whether addition of anthocyanins, which are strong scavengers of reactive oxygen species (ROS), improves development and intracellular redox status of heat-exposed bovine preimplantation embryos by reduction of heat shock-derived oxidative stress. After in vitro fertilization (IVF), embryos were cultured at 38.5 C through Day 8 (Day 0=day of IVF) with 0, 0.1, 1 and 10 microg/ml anthocyanins (non-heat-shocked group). On Day 2, embryos were cultured at 41.5 C for 6 h with 0, 0.1, 1 and 10 microg/ml anthocyanins followed by culture at 38.5 C until Day 8 (HS group). After exposure to heat shock, the intracellular ROS and glutathione (GSH) contents of individual embryos were measured in the non-heat-shocked and HS groups using fluorescent probes. On Day 8, the blastocysts formation rates of the embryos and total cell numbers of blastocysts were evaluated. Embryos exposed to heat shock without anthocyanins showed a significant decrease in blastocyst formation rate and GSH content (P<0.05) and an increase in intracellular ROS (P<0.05) compared with non-heat-shocked embryos. In contrast, addition of 0.1 microg/ml anthocyanins significantly (P<0.05) improved the blastocyst formation rate of the heat-shocked embryos. Addition of any dose of anthocyanins produced a significant decrease in the ROS levels (P<0.05) and tended to increase the GSH levels under heat-shock conditions. However, addition of higher concentrations (1 and 10 microg/ml) of anthocyanins to the culture media under heat shock did not improve the development of embryos. These results indicate that anthocyanins maintain the intracellular redox balance of heat-shocked bovine embryos by reducing intracellular oxidative stress and increasing the GSH levels. Thus, alterations of the redox state using natural antioxidative polyphenols is a useful approach for reducing heat shock-derived oxidative stress.     

Indole-3-carbaldehyde: a tyrosinase inhibitor from fungus YL185

Indole-3-carbaldehyde (1) was isolated as a tyrosinase inhibitor from the ethyl acetate-soluble fraction of extracellular fluids of unknown fungus YL185. The partial sequencing data of 18S ribosomal DNA (18S rDNA) indicate that this isolate belongs to the family Polyporaceae or Corticiaceae sensu lato. Indole-3-carbaldehyde inhibited the oxidation of l-3,4-dihydroxyphenylalanine (l-DOPA) by mushroom tyrosinase with a 50% inhibitory concentration (IC₅₀) of 1.3mM and showed inhibitory activity on melanin production in B16 melanoma cells. The aldehyde group of 1 plays an important role in eliciting tyrosinase inhibitory activity.     

Natural Resistance of Two Plantation Woods Populus × canadensis cv. and Cunninghamia lanceolata to Decay Fungi and Termites

Natural durability of two plantation woods, Chinese fir and I-214 poplar, was investigated thoroughly by three testing methods, namely an accelerated laboratory decay test, a fungus cellar test and a field test. Aft er the decay test using Postia placenta and Trametes versicolor, Chinese fir and the I-214 poplar showed 34% and 69% of mass loss, respectively, indicating they should be classified as slightly durable and non-durable wood. This conclusion was confirmed by the fungus cellar test and the field test. Like the performance in the decay test, I-214 poplar showed no resistance to termites either in the laboratory or in the field,whereas Chinese fir would be classified as moderately resistant.     

The first survey of Theileria orientalis infection in Mongolian cattle

In the present study, we have surveyed the presence of a bovine Theileria protozoan, Theileria orientalis, in Mongolian cattle and engorging tick populations from selected provinces and districts in Mongolia. The percentages of infection in the cattle and ticks ranged from 8.8 to 66.6 and from 3.7 to 73.3, respectively, on a per district basis. The genetic diversity of T. orientalis isolates was also studied, based on the protozoan gene encoding a major piroplasm surface protein (MPSP). At least five genotypes (types 1, 3, 5, 7, and N-3) of T. orientalis were found to be circulating among the Mongolian cattle and tick populations. In particular, types 3 and N-3 were common in most of the districts examined, while a strong geographical relationship among the genotypes was not detected in the present study. This is the first epidemiological report describing the presence of T. orientalis infection in Mongolian cattle.     

Gene expression profile of HIRRV G and N protein gene vaccinated Japanese flounder, Paralichthys olivaceus during HIRRV infection

The glycoprotein (G protein) gene, but not the nucleocapsid protein (N protein) gene, of the hirame rhabdovirus (HIRRV) was previously shown to be highly effective in inducing a protective immune response in Japanese flounder (Paralichthys olivaceus) when used as a DNA vaccine. Our previous cDNA microarray analysis demonstrated that interferon-stimulated genes (ISGs) were strongly induced by the HIRRV G protein gene (pHRV-G) but not by the N protein gene (pHRV-N). However, the molecular basis for the difference in protective immunity between pHRV-G- and pHRV-N-vaccinated fish during HIRRV infection is still unclear. In this study, we use a DNA microarray to analyze differences of gene expression in pHRV-G- and pHRV-N-vaccinated fish during HIRRV infection. Microarray analyses showed substantial difference in gene expression patterns during HIRRV infection between fish vaccinated with pHRV-G and pHRV-N. In addition, genes having homology to mammalian T cell activation-related genes were up-regulated in the HIRRV G protein-vaccinated group.