Evaluation of the effects of spores and their heat-treated residues from different Bacillus strains on the initial growth of rice plants

This study evaluated the effects of the spores and their heat-treated residues from different strains of Bacillus species (B. pumilus, B. altitudinis, and B. megaterium) on the early growth of paddy rice cultivars, including Hitomebore (short-grain japonica rice), Takanari (high-yielding indica rice), and two new lines, TULK-143-6 and LTAT-29. The spores of seven Bacillus strains positively affected Hitomebore root growth, while, the root volume of TULK-143-6 with inoculation of B. pumilus TUAT1 and JM52, and root length and root surface area of LTAT-29 with inoculation of B. megaterium MAFF301694 were increased significantly. In contrast with Hitomebore, Takanari root growth was significantly inhibited by the spores of six Bacillus strains. Surprisingly, inoculations with the spore residues from all tested Bacillus strains increased the root dry weight of Hitomebore, with the effects of four bacterial strains being significant. Furthermore, there were more Bacillus spores than vegetative cells at different time points during the initial rice growth stage, and most plant samples mainly consisted of Bacillus spores. Thus, the spores of Bacillus species likely promote rice root development.     

Isolation,characterization, and utilization of γ-aminobutyric acid (GABA)-producing lactic acid bacteria from Myanmar fishery products fermented with boiled rice

γ-Aminobutyric acid (GABA)-producing lactic acid bacteria (LAB) were isolated from four types of Myanmar traditional fermented fishery products with boiled rice. All of them belonged to the genus Lactobacillus, and comparison of the effects of these representatives on GABA accumulation in fermented fishery products with boiled rice revealed that Lactobacillus farciminis D323 is the most effective strain as a starter culture. These results may contribute to the development of traditional fermented fishery products with functional properties. In addition, this study is the first to show in detail the distribution of GABA-producing LAB in Southeast Asian fermented fishery products.     

Development of late-bolting F<Subscript>1</Subscript> hybrids of Chinese cabbage (<Emphasis Type="Italic">Brassica rapa</Emphasis> L.) allowing early spring cultivation without heating

We developed new F₁ hybrids of Chinese cabbage (Brassica rapa L. ssp. pekinensis) that allow cultivation earlier in spring without heating by introducing extremely late-bolting alleles at two homologs of the flowering repressor Flowering Locus C (BrFLC2 and BrFLC3) from non-heading ‘Leafy Green Parental Line No. 2’. These new F₁ hybrids were produced by the following four steps. First, the extremely late bolting selected lines were developed. These selected lines headed in spring after overwintering cultivation, whereas the conventional F₁ cultivars flowered. Secondly, an investigation of the three plantings showed that our F₁ hybrids formed heads when seeds were sown from mid-February to early March, whereas the conventional F₁ cultivar did not form heads because of premature bolting. Thirdly, we identified some F₁ hybrids with extremely late bolting during early spring cultivation in an investigation of many F₁ hybrids. Finally, based on an investigation across four cold regions for 2 years, we compared the commercialization rate, defined as the proportion of plants greater than 2000 g in weight and with a flowering stalk less than 10 cm long. Then we identified a F₁ of MS02 × 12-04 which had a high commercialization rate on average (92%), whereas the rates of three conventional F₁ cultivars were only 0–2%. In the near future, these F₁ hybrids will be valuable late-bolting cultivars despite climate change, permitting stable cultivation and harvest over wide regions.     

Identification and some properties of anthocyanin isolated from Zuiki, stalk of Colocasia esculenta

Zuiki, a stalk of taro (Colocasia esculenta), is a traditional vegetable in Japan. Raw zuiki is often boiled and vinegared to eat. The surface color of zuiki is reddish. Here, we isolated a red pigment from zuiki and identified it as cyanidin 3-rutinoside using instrumental analyses. The color of zuiki disappeared by boiling, but the zuiki turned red again in an acetic acid solution. It seems that the cyanidin 3-rutinoside that exists on the surface of zuiki elutes in boiling water and then, the pigment that seeps out from the inside of the zuiki is exposed to an acid solution, and its surface turns red again. The radical scavenging activity of purified zuiki anthocyanin was 114 mg equivalent to BHT/g. About half of the anthocyanin in fresh zuiki was washed out by boiling, and the radical scavenging activity of zuiki was definitely reduced.     

Comparison of extractive components in muscle and liver of three Loliginidae squids with those of one Ommastrephidae species

Extractive components were determined for the mantle muscle and liver of three species of Loliginidae squid, Sepioteuthis lessoniana, Loligo bleekeri, and L. edulis, and one species of Ommastrephidae, Todarodes pacificus, as a control. Total free amino acid levels and the major free amino acids in muscle, taurine, proline, glycine, alanine, and arginine, were significantly higher in Loliginidae squids than in T. pacificus. The main nucleotide was adenosine 5′-monophosphate, which did not differ significantly between species. Malate was the organic acid found in muscle in highest concentrations. The muscle of these species contained a large amount of trimethylamine oxide. A large amount of glycine betaine was also detected in the muscle, but showed no large species difference. From these results, the muscle of Loliginidae species is considered to have a much sweeter taste than that of T. pacificus. Compared with muscle, squid liver was characteristic with high contents of taurine, glutamate, bitter amino acids, succinate, propionate, trimethylamine, and glycine betaine, and with low contents of sweet amino acids, arginine, nucleotides, malate, and trimethylamine oxide. These results suggest that squid liver is characterized by a complicated taste containing umami, bitterness, sourness, fishy flavor, and less sweetness.     

Relationship between the timing of insemination based on estrus detected by the automatic activity monitoring system and conception rates using sex-sorted semen in Holstein dairy cattle

We investigated the optimal timing of artificial insemination (AI) for achieving pregnancy according to the onset/end of estrus detected by an accelerometer system in Holstein cattle. The conception rates of conventional semen were used as a reference. The conception rate from AI of sex-sorted semen was higher at −4 to 4 h (57.1%) from the end of estrus than those at −12 to −4 h (37.7%) and 12–20 h (30.3%), whereas AI at 4–12 h showed an intermediate conception rate (47.4%). Conversely, conception rates were similar in AI performed between 0 and 32 h from the onset of estrus. Regarding conventional semen, the interval from the onset and end of estrus did not affect conception rates. The present results suggest that the time of the end of estrus is the better indicator of optimal AI timing for sex-sorted semen than the onset of estrus.     

Immunoblot analysis of specific antigen bands predictable for Dirofilaria immitis infection in cats

Serial sera from four mongrel cats experimentally inoculated with infectious larvae of Dirofilaria immitis were analyzed by immunoblot patterns against a phosphate buffered saline-extract of D. immitis. Antigen-specific protein bands detected indicate that the low molecular weight bands of 36, 32, 22, 19 and 14 kDa, are predictable for positive adult worm infection, suggesting diagnostic usefulness for adult D. immitis infection in cats.     

Regulation mechanism of selective atresia in porcine follicles: regulation of granulosa cell apoptosis during atresia

More than 99% of follicles undergo a degenerative process known as "atresia", in mammalian ovaries, and only a few follicles ovulate during ovarian follicular development. We have investigated the molecular mechanism of selective follicular atresia in mammalian ovaries, and have reported that follicular selection dominantly depends on granulosa cell apoptosis. However, we have little knowledge of the molecular mechanisms that control apoptotic cell death in granulosa cells during follicle selection. To date, at least five cell death ligand-receptor systems [tumor necrosis factor (TNF)alpha and receptors, Fas (also called APO-1/CD95) ligand and receptors, TNF-related apoptosis-inducing ligand (TRAIL; also called APO-2) and receptors, APO-3 ligand and receptors, and PFG-5 ligand and receptors] have been reported in granulosa cells of porcine ovaries. Some cell death ligand-receptor systems have "decoy" receptors, which act as inhibitors of cell death ligand-induced apoptosis in granulosa cells. Moreover, we showed that the porcine granulosa cell is a type II apoptotic cell, which has the mitochondrion-dependent apoptosis-signaling pathway. Briefly, the cell death receptor-mediated apoptosis signaling pathway in granulosa cells has been suggested to be as follows. (1) A cell death ligand binds to the extracellular domain of a cell death receptor, which contains an intracellular death domain (DD). (2) The intracellular DD of the cell death receptor interacts with the DD of the adaptor protein (Fas-associated death domain: FADD) through a homophilic DD interaction. (3) FADD activates an initiator caspase (procaspase-8; also called FLICE), which is a bipartite molecule, containing an N-terminal death effector domain (DED) and a C-terminal DD. (4) Procaspase-8 begins auto-proteolytic cleavage and activation. (5) The auto-activated caspase-8 cleaves Bid protein. (6) The truncated Bid releases cytochrome c from mitochondrion. (7) Cytochrome c and ATP-dependent oligimerization of apoptotic protease-activating factor-1 (Apaf-1) allows recruitment of procaspase-9 into the apoptosome complex. Activation of procaspase-9 is mediated by means of a conformational change. (8) The activated caspase-9 cleaves downstream effector caspases (caspase-3). (9) Finally, apoptosis is induced. Recently, we found two intracellular inhibitor proteins [cellular FLICE-like inhibitory protein short form (cFLIPS) and long form (cFLIPL)], which were strongly expressed in granulosa cells, and they may act as anti-apoptotic/survival factors. Further in vivo and in vitro studies will elucidate the largely unknown molecular mechanisms, e. g. which cell death ligand-receptor system is the dominant factor controlling the granulosa cell apoptosis of selective follicular atresia in mammalian ovaries. If we could elucidate the molecular mechanism of granulosa cell apoptosis (follicular selection), we could accurately diagnose the healthy ovulating follicles and precisely evaluate the oocyte quality. We hope that the mechanism will be clarified and lead to an integrated understanding of the regulation mechanism.