Effects of acorn attack by curculio weevils on the germination and early growth of <Emphasis Type="Italic">Pasania edulis</Emphasis> (Makino) seedlings

We conducted field studies to evaluate the impact that curculio weevil attacks on Pasania edulis (Fagacea) acorns have on the regeneration of this tree. The germination ratio of weevil-attacked acorns was significantly lower than that of sound acorns. The number and position of the attacks on acorns affected the germination ratio. The seedlings from weevil-attacked acorns were shorter and had a smaller leaf area than those from sound acorns. The negative effects on seedling height lasted for at least three years after germination. We also studied the handling of the two types of acorns by granivorous Apodemus mice in a broad-leaved forest dominated by P. edulis. The mice picked up, transported, hoarded, and recovered sound and weevil-attacked acorns similarly. In the study site, the seedlings from weevil-attacked acorns were estimated to account for 1.5–20.4 % of the total seedlings for cohorts of mast years and 0–3.7 % for those of poor or medium acorn production. From these results, we conclude that acorns with the lower part of their cotyledons slightly damaged by weevil larvae might still be able to contribute to the regeneration of P. edulis in the field but that their contribution would be negligible or small even in mast years in this study site.     

Uniquely stable 40 kDa subfragment-2 in carp myosin

Digestion of carp myofibrils at 30 degrees C in 0.5 M KCl medium with calcium ion generated unique 135 kDa heavy meromyosin (HMM). The HMM was not produced when digested at 10 degrees C. A further digestion of the 135 kD HMM isolated in the absence of calcium ion generated uniquely short subfragment-2 (S-2) with a size of 40 kDa (40 kDa S-2) together with subfragment-1 (S-1). The 40 kDa S-2 was identified by N- and C-end sequencing, and demonstrated to locate the amino end of the rod portion. The unfolding temperature for the 40 kDa S-2 was around 52 degrees C as studied by circular dichroism measurement. The same unfolding peak was also detected with the intact rod together with a large unfolding peak at around 36 degrees C coming from the rest of the rod portion, light meromyosin. The unfolding peak for the 40 kDa S-2 in myosin was a little lower (48 degrees C) than that in free form, suggesting the involvement of the head portion in the stability of the 40 kDa S-2 in the structure.     

Isolation and characterization of microorganisms capable of hydrolysing the herbicide mefenacet

Three strains—Nocardioides sp. MFC-A, Rhodococcus rhodochrous MFC-B, and Stenotrophomonas sp. MFC-C—were newly isolated from rice soil or bark compost as microorganisms capable of cometabolically degrading mefenacet (2-benzothiazol-2-yloxy-N-methyl-acetanilide). Several other species belonging to the genera Nocardioides, Rhodococcus, and Stenotrophomonas were also capable of degrading mefenacet, suggesting that the ability is not specific to these isolates, but common at least in these genera. N-methylaniline and 2-benzothiazoloxy acetic acid were identified as the metabolites by liquid chromatography-mass spectrometry. Besides mefenacet, the isolates could degrade one or more other amide pesticides, such as flutolanil, mepronil, metolachlor, and pretilachlor. These results indicate that the strains Nocardioides sp. MFC-A, Rhodococcus rhodochrous MFC-B, and Stenotrophomonas sp. MFC-C degrade mefenacet via hydrolysis of the amide bond, but their substrate specificities differ.     

Identification of a potent immunostimulatory oligodeoxynucleotide from Streptococcus thermophilus lacZ

Immunostimulatory sequences of oligodeoxynucleotides (ODNs), such as CpG ODNs, are potent stimulators of innate immunity. Here, we identified a strong immunostimulatory CpG ODN, which we named MsST, from the lac Z gene of Streptococcus (S.) thermophilus ATCC19258, and we evaluated its immune functions. In in vitro studies, MsST had a similar ability as the murine prototype CpG ODN 1555 to induce inflammatory cytokine production and cell proliferation. In mouse splenocytes, MsST increased the number of CD80+CD11c+and CD86+CD11c+ dendritic cells and CD4+CD25+ regulatory T cells. We also analyzed the effects of MsST on the expression of regulatory cytokines by real-time quantitative PCR. MsST was more potent at inducing interleukin-10 expression than the ODN control 1612, indicating that MsST can augment the regulatory T cell response via Toll-like receptor 9, which plays an important role in suppressing T helper type 2 responses. These results suggest that S. thermophilus , whose genes include a strong Immunostimulatory sequence-ODN, is a good candidate for a starter culture to develop new physiologically functional foods and feeds.     

Microbiota analysis of Caspian Sea yogurt, a ropy fermented milk circulated in Japan

We analyzed the microbiota of domestic ropy fermented milk, Caspian Sea yogurt (or 'kasupikai yohguruto' in Japanese), circulated in Japan. We collected six varieties from five localities. Lactococcus ( L) lactis ssp. cremoris was isolated from all samples as the dominant strain at levels of 10⁸−10⁹ CFU/g. We show this strain produces an extracellular polysaccharide (EPS) that causes the unique characteristic viscosity of the product. From analysis of the RAPD pattern of 60 bacterial isolates from the six samples, we found that 59 strains from a total of 60 isolates were identical and produced this viscosity. Furthermore, PFGE analysis of representative strains from each sample indicated that the isolates could be classified into four subgroups. This suggests these L. lactis ssp. cremoris strains found in Caspian Sea yogurt may have been slightly mutated during subculture in Japan. In addition, Lactobacillus (L.) sakei ssp. sakei were isolated from three samples; L. plantarum , Gluconoacetobacter sacchari and Acetobacter aceti were isolated from two samples; and L. paracasei , L. kefiri , Leuconostoc ( Leu. ) mesenteroides were isolated from one sample.     

Identification of the core rumen bacterial taxa and their population dynamics during the fattening period in Japanese Black cattle

The rumen microbiota comprises a vast range of bacterial taxa, which may affect the production of high-quality meat in Japanese Black cattle. The aim of this study was to identify core rumen microbiota in rumen fluid samples collected from 74 Japanese Black cattle raised under different dietary conditions using 16S rRNA gene amplicon sequencing. In the rumen of fattening Japanese Black cattle, 10 bacterial taxa, showing >1% average relative abundance and >95% prevalence, irrespective of the dietary conditions and the fattening periods, were identified as the core rumen bacterial taxa, which accounted for approximately 80% of the rumen microbiota in Japanese Black cattle. Additionally, population dynamics of the core rumen bacterial taxa revealed two distinct patterns: Prevotella spp. and unclassified Bacteroidales decreased in the mid-fattening period, whereas unclassified Clostridiales, unclassified Ruminococcaceae, Ruminococcus spp., and unclassified Christensenellaceae increased during the same period. Therefore, the present study reports the wide distribution of the core rumen bacterial taxa in Japanese Black cattle, and the complementary nature of the population dynamics of these core taxa, which may ensure stable rumen fermentation during the fattening period.     

Effects of a probiotic bacterial <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> dietary supplement on the crowding stress response of juvenile Nile tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

To test the effects of probiotic bacteria against crowding stress, juvenile Nile tilapia Oreochromis niloticus were fed commercial feed supplemented with 1 × 10¹⁰ cfu/g pellets of Lactobacillus rhamnosus for 3 weeks. The fish were confined and subjected to crowding for 7 and 14 days after which their physiological condition was investigated. The administration of probiotic prophylaxis improved growth performance even in the stressed fish. After 7 days of stress exposure, fish receiving probiotic-supplemented feed showed proactive behavior and coping responses to the stressor, as evidenced by elevated plasma glucose levels and osmolality, stabilized plasma electrolytes, and a higher RNA:DNA ratio. Fish stressed for 7 days and fed normal commercial diet showed impairment of plasma electrolytes; after 14 days, the plasma osmolality, electrolytes, glucose, and RNA:DNA ratio decreased, indicating physiological maladaptation to the stressor. The growth rate was also found to be reduced, suggesting that energy demand exceeded the energy available from metabolism and dietary uptake. This latter effect was not observed in fish fed the probiotic-supplemented diet. The results suggest that probiotic bacteria prophylaxis may increase energy availability for metabolic support of the crowding stress response and improve the stress coping capacity of fish.     

Effect of ensiled hop (Humulus lupulus L.) residues on plasma acetate turnover rate in sheep

An isotope dilution method using [1‐¹³C]sodium acetate was applied to determine the effect of feeding ensiled hop (Humulus lupulus L.) residues on plasma acetate turnover rate in six adult crossbred sheep. The sheep were fed 63 g/kg body weight (BW)^0.75/day of either mixed hay of orchardgrass (Dactylis glomerata L.) and reed canarygrass (Phalaris arundinacea L.) and round bale silage at 3:1 ratio (Hay‐diet), or another where round bale silage was replaced by ensiled hop residues (Hop‐diet) with a crossover design each of a 3‐week period. The isotope dilution method was performed on day 21 of each dietary treatment. Dry matter digestibility was similar between diets, and nitrogen (N) digestibility was lower (P = 0.001) for Hop‐diet than Hay‐diet. However, N retention did not differ between diets. Plasma acetate concentration was lower (P = 0.04) for Hop‐diet than Hay‐diet, and the turnover rate of plasma acetate did not differ between diets. Plasma concentration of lactate and non‐esterified fatty acids were similar between diets. Hop‐diet was found almost comparable to Hay‐diet on plasma acetate turnover rate in the present experimental conditions. Therefore, it could be concluded that hop residues partially could be used as an alternative to traditionally used round bale silage for rearing sheep.     

DNA methylation analysis on satellite I region in blastocysts obtained from somatic cell cloned cattle

Many observations have been made on cloned embryos and on adult clones by somatic cell nuclear transfer (SCNT), but it is still unclear whether the progeny of cloned animals is presenting normal epigenetic status. Here, in order to accumulate the information for evaluating the normality of cloned cattle, we analyzed the DNA methylation status on satellite I region in blastocysts obtained from cloned cattle. Embryos were produced by artificial insemination (AI) to non‐cloned or cloned dams using semen from non‐cloned or cloned sires. After 7 days of AI, embryos at blastocyst stage were collected by uterine flushing. The DNA methylation levels in embryos obtained by using semen and/or oocytes from cloned cattle were similar to those in in vivo embryos from non‐cloned cattle. In contrast, the DNA methylation levels in SCNT embryos were significantly higher (P < 0.01) than those in in vivo embryos from non‐cloned and cloned cattle, approximately similar to those in somatic cells used as donor cells. Thus, this study provides useful information that epigenetic status may be normal in the progeny of cloned cattle, suggesting the normality of germline cells in cloned cattle.