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A lactic acid bacterium, Lactiplantibacillus plantarum P1-2 (LpP1-2), can reduce environmental fatty acid hydroperoxides. The administration of LpP1-2 to oxygen-sensitive short-lived nematode mutants and iron-overloaded rats reduced the oxidative stress-related index. Since young piglets have a weak defense system against oxidative stress and are vulnerable to environmental stress, antimicrobial agents have been administered in the rearing. Based on these results, we investigated the effect of LpP1-2 administration to prepartum sows and infant piglets until weaning without antimicrobial agents on the growth of young piglets.The group including both sows and piglets that were administrated with lactic acid bacteria containing LpP1-2 (LABLp) until the end of lactation showed the growth-promoting effect of piglet from lactation to early regular rearing, and even in late regular rearing. Blood biochemical markers were in the normal ranges in both LABLp-administrated and non-administrated groups, but various disease-related markers tended to decrease in the administrated group.To investigate the effects of LABLp administration on postpartum piglets, the piglets born from prenatally LABLp-administrated sows were divided into two groups and then administrated with or without LABLp. The piglets in the LABLp-administrated group tended to grow very slightly higher than those in the non-administrated group from lactation to early regular rearing. After that, the growth in both groups was almost the same. These results suggest that LABLp administration to prepartum sows is essential for the growth-promoting effect.The postnatal LABLp-administrated piglets showed a lower serum lipid peroxidation index than the non-administrated piglets, and had higher numbers of lactic acid bacteria and bifidobacteria in feces at the end of LABLp treatment. In fattening performances, the LABLp-administrated group showed a significant improvement in meat quality.We also discuss the growth and physiological effects by LpP1-2 administration with iron on piglets because iron administration is another important issue in piglet farming.

Fourteen novel lipomycetaceous yeasts species were isolated from soil samples collected from the Hokkaido, Chiba and Okinawa prefectures of Japan. Phylogenetic analyses of the D1/D2 domains of the large subunit rRNAs and translation elongation factor 1 alpha genes (TEF1-α) revealed that five strains of two species from the soil in Furano-shi, Hokkaido were related to Dipodascopsis anomala and 29 strains representing 12 species from soils in Kamogawa-shi, Chiba and Iriomote Island, Okinawa were in the Myxozyma clade. The two species of Dipodascopsis form globose or ellipsoid ascospores in their sac-like ascus and pseudohyphae. Furthermore, these species produce ascospores in their pseudohyphae and do not produce an acicular ascus, which is common among the three species including D. anomala. Therefore, we propose transferring D. anomala to the genus Babjevia and amending Babjevia. Two novel species were described and included in the genus Babjevia: Babjevia hyphoforaminiformans sp. nov. (holotype NBRC 111233; MycoBank no. MB 829051) and Babjevia hyphasca sp. nov. (holotype NBRC 112965; MycoBank no. MB 829053). The 12 species in the Myxozyma clade produce neither ascospores nor pseudohyphae and have different characteristics in assimilating several carbon sources from each other. Thus, we propose that the novel species of Lipomyces be classified as forma asexualis (f.a.). From Kamogawa-shi, Chiba (19 strains representing five species): Lipomyces melibiosiraffinosiphilus f.a., sp. nov. (holotype NBRC 111411; MycoBank no. MB 829034), Lipomyces kiyosumicus f.a., sp. nov. (holotype NBRC 111424; MycoBank no. MB 829035), Lipomyces chibensis f.a., sp. nov. (holotype NBRC 111413; MycoBank no. MB 829036), Lipomyces kamogawensis f.a., sp. nov. (holotype NBRC 112967; MycoBank no. MB 829037), Lipomyces amatsuensis f.a., sp. nov. (holotype NBRC 111420; MycoBank no. MB 829041). From Iriomote island, Okinawa (10 strains representing seven species): Lipomyces taketomicus f.a., sp. nov. (holotype NBRC 112966; MycoBank no. MB 829042), Lipomyces yaeyamensis f.a., sp. nov. (holotype NBRC 110433; MycoBank no. MB 829050), Lipomyces iriomotensis f.a., sp. nov. (holotype NBRC 110436; MycoBank no. MB 829045), Lipomyces haiminakanus f.a., sp. nov. (holotype NBRC 110435; MycoBank no. MB 829046), Lipomyces komiensis f.a., sp. nov. (holotype NBRC 110440; MycoBank no. MB 829047), Lipomyces nakamensis f.a., sp. nov. (holotype NBRC 110434; MycoBank no. MB 829048), Lipomyces sakishimensis f.a., sp. nov. (holotype NBRC 110439; MycoBank no. MB 829049).

Fourteen novel lipomycetaceous yeasts species were isolated from soil samples collected from the Hokkaido, Chiba and Okinawa prefectures of Japan. Phylogenetic analyses of the D1/D2 domains of the large subunit rRNAs and translation elongation factor 1 alpha genes (

Nine bacterial strains were isolated from Thai fermented fish by the standard dilution technique using JCM no. 377 medium. The rod shaped isolate FN2-3 was identified as Virgibacillus dokdonensis while isolates FN2-3 and FN6-6 as V. halodenitrificans, FN1-13 as Corynebacterium variabile, FN1-10 as Oceanobacillus iheyensis and FN3-7 was Bacillus amyloliquefaciens subsp. plantarum. The coccal isolates, FN6-1 and FN6-7 were Staphylococcus saprophyticus subsp. bovis and FN6-8 was S. saprophyticus subsp. saprophyticus, based on their phenotypic characteristics and 16S rRNA gene sequence analyses at a 99.63-100% sequence similarity. Their lipase activity in complex medium (CM), CM medium with 1% (v/v) Tween 20 or CM medium with 1% (v/v) Tween 80 ranged from 1.12 ± 0.03-3.77 ± 0.04 unit/mL, with the highest lipase activity found with V. dokdonensis FN1-8 cultivated with CM with 1% (v/v) Tween 80.

Reinforcement of hydroperoxide-eliminating activity in the intestines and colon should prevent associated diseases. We previously isolated a lactic acid bacterium,Pediococcus pentosaceusBe1, that facilitates a 2-electron reduction of hydrogen peroxide to water. In this study, we successfully isolated an alternative lactic acid bacterium,Lactobacillus plantarumP1-2, that can efficiently reduce environmental alkyl hydroperoxides and fatty acid hydroperoxides to their corresponding hydroxy derivatives through a 2-electron reduction. Each strain exhibited a wide concentration range with regard to the environmental reducing activity for each hydroperoxide. Given this, the two lactic acid bacteria were orally administered to the oxygen-sensitive short-lived nematode mutant, and this resulted in a significant expansion of its lifespan. This observation suggests thatP. pentosaceusBe1 andL. plantarumP1-2 inhibit internal oxidative stress. To determine the specific organs involved in this response, we performed a similar experiment in rats, involving induced lipid peroxidation by iron-overloading. We observed that onlyL. plantarumP1-2 inhibited colonic mucosa lipid peroxidation in rats with induced oxidative stress.

A non-toxigenic mutant of the toxigenic serotype C Clostridium botulinum strain Stockholm (C-St), C-N71, does not produce the botulinum neurotoxin (BoNT). However, the original strain C-St produces botulinum toxin complex, in which BoNT is associated with non-toxic non-hemagglutinin (NTNHA) and three hemagglutinin proteins (HA-70, HA-33, and HA-17). Therefore, in this study, we aimed to elucidate the effects of bont gene knockout on the formation of the "toxin complex." Nucleotide sequence analysis revealed that a premature stop codon was introduced in the bont gene, whereas other genes were not affected by this mutation. Moreover, we successfully purified the "toxin complex" produced by C-N71. The "toxin complex" was identified as a mixture of NTNHA/HA-70/HA-17/HA-33 complexes with intact NTNHA or C-terminally truncated NTNHA, without BoNT. These results indicated that knockout of the bont gene does not affect the formation of the "toxin complex." Since the botulinum toxin complex has been shown to play an important role in oral toxin transport in the human and animal body, a non-neurotoxic "toxin complex" of C-N71 may be valuable for the development of an oral drug delivery system.

To obtain lactic acid bacteria that scavenge environmental hydrogen peroxide, we developed a specialized enrichment medium and successfully isolated Pediococcus pentosaceus Be1 strain from a fermented food. This strain showed vigorous environmental hydrogen peroxide scavenging activity over a wide range of hydrogen peroxide concentrations. High Mn-catalase and NADH peroxidase activities were found in the cell-free extract of the P. pentosaceus Be1 strain, and these two hydrogen peroxide scavenging enzymes were purified from the cell-free extract of the strain. Mn-catalase has been purified from several microorganisms by several researchers, and the NADH peroxidase was first purified from the original strain in this report. After cloning the genes of the Mn-catalase and the NADH peroxidase, the deduced amino acid sequences were compared with those of known related enzymes.

Thai traditional fermented shrimp paste (Ka-pi) is widely consumed as condiment and seasoning ingredient in Thailand. Chemical and physical properties of thirteen shrimp paste samples were examined. The water content and water activity (Aw) of the products ranged from 33.95-52.19 % and 0.64-0.72, respectively while salt concentration was 7.00 to 10.85 %. The nitrogen content varied from 2.87 to 6.85 % (w/w). KS1 showed the highest protein content (42.8 %) followed by SP1 and SP3 (42.4 and 41.3 %), respectively. The predominant amino acids were glutamic acid (70.1-593.9 µg/g), lysine (112.7-546.3 µg/g) and leucine (29.5-544.9 µg/g). Total bacterial cell count in the samples ranged from 1.3x103 - 2.9x105 cfu g-1 while lactic acid bacteria (LAB) were not detected. This study will be provided the information for improving the quality of products in manufacture to meet the demand of consumer.

Background For internal fixation of AO classification Type B lateral malleolar fracture, insertion of lag screws into the fracture plane and fixation with a one-third tubular plate as a neutralization plate are the standard treatment procedures. The one-third tubular plate is processed to a hook shape and hung on the distal end of the fibula. In this study, to compare the function of the hook and lag screws of a one-third tubular plate and LCP for osteosynthesis of lateral malleolar fracture, mechanical indices of internal fixation were compared among the one-third tubular plates with lag screws with and without the hook and a locking compression plate. Methods As mechanical tests, a compression test was performed in which compression in the bone axis direction produced by supporting the body weight was simulated, and a torsion test was performed in which external rotation of the bone axis caused by plantar flexion of the ankle joint was simulated. Muscle strength during walking and the force and torque acting on the ankle and knee joints were determined using inverse dynamic analysis. Finite element analysis was performed to analyze the function of hooks and lag screws. The joint reaction force determined by inverse dynamic analysis was adopted as the loading condition of finite element analysis. Results A stiffness equivalent to that of healthy bone could be achieved by all three internal fixations. It was clarified that the presence of the hook does not make a difference in stiffness. Displacement of the one-third tubular plate was small regardless of the presence or absence of the hook compared with those of locking compression plates. Conclusions The presence of the hook did not make any difference in stiffness, suggesting that active preparation of the hook is unnecessary. We also clarified that lag screws inhibit displacement.

Amylase isozymes from non-glutinous japonica and indica rice types showed various band patterns, strengths and intensities based on zymography. Major bands were presumed to be α- and β-amylase according to their molecular weights; other amylases, 270 and 200 kDa in size, were also detected. Three debranching enzymes (DBEs) were detected from the dye pattern as well. Amylase activities were observed at pH 7.0 during rice cooking. However, the α-amylase activity at pH 7.0 was 50% lower than that at pH 5.0; DBE, β-amylase, and α-glucosidase values were similarly reduced by 25%, 90% and 99%, respectively. α-Amylase activity and DBE activities correlated positively with both the amount of maltooligomer leached during rice cooking and the adhesiveness of cooked rice. Thus, it is considered that the activity of amylase isozymes affects the palatability of cooked rice. Therefore, enzymes from rice endosperm may represent a new and highly useful biomarker for evaluating the palatability of cooked rice.

Objective We have conducted a basic study on the influences on ultrasonic properties when LIPUS is applied through wound dressing. According to the results of ex vivo experiments conducted to date, LIPUS showed ultrasonic properties such as transmittance, coefficient of transmission, and a non-uniformity ratio through film wound dressing better than other wound dressing, and it was considered that LIPUS's effect for fracture healing was not influenced by film wound dressing. Then, we discussed the influence on the effect of LIPUS through film wound dressing. Methods Thirty male 8-week-old Sprague-Dawley rats were used for the trial. After creating close transverse femoral fractures on the right legs of these 30 rats, they were divided into 3 groups of 10; LIPUS through wound dressing (Group A), LIPUS without wound dressing (Group B), and No LIPUS treatment (Group C). OPSITE Wound, which was thought to have the least influence on ultrasound properties, was used for this trial. Group A and B received LIPUS for 20 minutes a day from the first day after the fractures. LIPUS was generated from Teijin Pharma's device for a basic experiment. When treating Group A, the wound dressing was pasted on the ultrasound terminal in order to apply LIPUS through the dressing. We assessed the time-oriented morphological change of each group in anesthetized condition using simple radiographs on the 8th, 16th, and 24th day after the fractures. Results Six rats in Group A, 2 in Group B, and 1 in Group C died in anesthesia, and we discussed the remaining 4 rats in Group A, 8 in Group B, and 9 in Group C. We defined more than one teleost callus bridging as bone-union. We also counted a bone remodeling when we recognized the absorption of existing cortical bone and the transformation of new bone to cortical bone in simple radiographs. As a result, compared with Group C, we recognized that both bone union and remodeling accelerated remarkably in Group B, but not in Group A. Discussion It suggested that LIPUS through wound dressing had negative influences on both period shorting of fracture healing and bone remodeling. When LIPUS was conducted through film wound dressing, transmittance and coefficient of transmission were unchanged; however, the non-uniformity ratio changed slightly. The non-uniformity ratio of the ultrasound transducer had a significant influence on the effect of LIPUS on fracture healing.

Botulinolysin (BLY) is a toxin produced by Clostridium botulinum that belongs to a group of thiol-activated hemolysins. In this study, a protein exhibiting hemolytic activity was purified from the culture supernatant of C. botulinum serotype D strain 4947. The purified protein displayed a single band by sodium dodecyl sulfate polyacrylamide gel electrophoresis with a molecular mass of 55 kDa, and its N-terminal and internal amino acid sequences exhibited high similarity to a group of thiol-activated hemolysins produced by gram-positive bacteria. Thus, the purified protein was identified as the BLY. Using the nucleotide sequences of previously cloned genes for hemolysins, two types of genes encoding BLY-like proteins were cloned unexpectedly. Molecular modeling analysis indicated that the products of both genes displayed very similar structures, despite the low sequence similarity. In silico screening revealed a specific duplication of the hemolysin gene restricted to serotypes C and D of C. botulinum and their related species among thiol-activated hemolysin-producing bacteria. Our findings provide important insights into the genetic characteristics of pathogenic bacteria.

Phospholipase D (PLD) catalyzes transphosphatidylation, causing inter-conversion of the polar head group of phospholipids and phospholipid hydrolysis. Previously, we cloned PLD103, a PLD with high transphosphatidylation activity, from Streptomyces racemochromogenes strain 10-3. Here, we report the construction of an expression system for the PLD103 gene using Streptomyces lividans as the host bacterium to achieve large-scale production. The phosphatidylcholine (PC) hydrolysis activity of S. lividans transformed with the expression plasmid containing the PLD103 gene was approximately 90-fold higher than that of the original strain. The recombinant PLD103 (rPLD103) found in the supernatant of the transformant culture medium was close to homogeneous. The rPLD103 was indistinguishable from the native enzyme in molecular mass and enzymatic properties. Additionally, rPLD103 had high transphosphatidylation activity on PC as a substrate in a simple aqueous one-phase reaction system and was able to modify the phospholipid content of soybean lecithin. Consequently, the expression system produces a stable supply of PLD, which can then be used in the production of phosphatidyl derivatives from lecithin.

The emu (Dromaius novaehollandiae) egg is considered promising as an alternative egg product. To obtain basic biochemical information on emu egg white, the major protein compositions in emu and chicken egg whites and the primary structures of potential allergenic proteins were compared. The dominant protein in emu egg white was ovotransferrin (OVT), followed by ovalbumin (OVA) and TENP protein. The OVA and ovomucoid (OVM) levels in emu egg white were estimated as significantly lower than those in chicken egg white by Western blotting and enzyme-linked immunosorbent assays using anti-chicken OVA or OVM antibodies. Lysozyme and its enzymatic activity were not detected in emu egg white. OVT, OVA, and OVM genes were also cloned, and their nucleotide and amino acid sequences were determined. The protein sequences of OVT, OVA, and OVM from emu showed lower similarities to those of chicken than other avian species, such as quail and turkey. These results emphasize the low allergenicity of emu egg white and its potential as an alternative to chicken egg white.