Polyethylene degradation by lignin-degrading fungi and manganese peroxidase

Degradation of high-molecular-weight polyethylene membrane by lignin-degrading fungi, IZU-154, Phanerochaete chrysosporium, and Trametes versicolor, was investigated under various nutritional conditions. IZU-154 showed the most significant polyethylene degradation among the three lignin-degrading fungi under nitrogen- or carbon-limited culture conditions. Furthermore, for T. versicolor and P. chrysosporium, the addition of Mn(II) into nitrogen- or carbon-limited culture medium enhanced polyethylene degradation. These results suggest that polyethylene degradation is related to ligninolytic activity of lignin-degrading fungi. Treatment of polyethylene membrane with partially purified manganese peroxidase (MnP) caused significant degradation in the presence of Tween 80, Mn(II), and Mn(III) chelator. This result demonstrates that MnP is the key enzyme in polyethylene degradation by lignin-degrading fungi.This study was presented in part at the 9th International Symposium on Wood and Pulping Chemistry, Montreal, Canada, June 9–12, 1997     

Relationship between prey utilization and growth variation in hatchery‐reared Pacific bluefin tuna,Thunnus orientalis (Temminck et Schlegel), larvae estimated using nitrogen stable isotope analysis

In mass culture of Pacific bluefin tuna Thunnus orientalis, a marked growth variation is observed after they start feeding at 6–7 mm in body length (BL) on yolk‐sac larvae of other species, and the growth variation in tuna larvae is a factor leading to the prevalence of cannibalism. To examine the relationship between prey utilization and growth variation, nitrogen stable isotope ratios (δ¹⁵N) of individual larvae were analysed. A prey switch experiment was conducted under two different feeding regimes: a group fed rotifers (rotifer fed group), and a group fed yolk‐sac larvae of spangled emperor, Lethrinus nebulosus (fish fed group) from 15 days after hatching (6.87 mm BL). The fish fed group showed significantly higher growth than the rotifer fed group. Changes in the δ¹⁵N of the fish fed group were expressed as an exponential model and showed different patterns from those of the rotifer fed group. The δ¹⁵N of fast‐growing tuna larvae collected in an actual mass culture tank after the feeding of yolk‐sac larvae was significantly higher than those of the slow‐growing larvae, indicating that slow glowing larvae depended largely on rotifers rather than the yolk‐sac larvae.     

Encephalomalacia with Enterococcus durans infection in the brain stem and cerebral hemisphere in chicks in Japan

Fourteen chicks, 7-12 days old, that died or had neurological signs (depression, paralysis, torticollis, incoordination, and rolling) were investigated pathologically and microbiologically. Variably sized foci of malacia were present within the parenchyma of the brain stem (cerebral peduncle, optic lobe, and medulla oblongata) and the cerebral hemisphere. Capillary thrombosis with congestion and hemorrhage was frequently observed within the malacia lesions. Gram-positive cocci were recognized in the blood capillary of malacia lesions. Bacteriologically, Enterococcus durans was isolated from the brain, liver, kidney, lung, and spleen. Detection of gram-positive cocci within the blood vessels in the malacia lesions observed in these cases may more strongly suggest the pathogenic role of E. durans on the malacia lesions in the brain stem and cerebral hemisphere.     

Periorbital cyst with bone defect in a dog

A 4-year-old female Miniature Dachshund was referred with a chief complaint of right periorbital swelling that had not responded to antibiotic therapy. Ultrasonography and fine-needle aspiration revealed that the periorbital lesion had a cystic structure without any inflammatory or neoplastic cells. Computed tomography (CT) showed that the cyst occupied a defect in the periorbital maxillary, lacrimal, and frontal bones and had invaded the nasal cavity. The lesion was histologically suspected by incisional biopsy as an epithelial cyst.     

Glutathione supplementation to semen extender improves the quality of frozen-thawed canine spermatozoa for transcervical insemination

The present study was conducted to evaluate whether supplementation of semen extender with glutathione (GSH) can maintain the quality of frozen-thawed canine spermatozoa. Eighteen ejaculates were obtained from 5 dogs and placed in extender (20% egg yolk, Tris, citric acid, lactose, raffinose, antibiotics and 6.5% glycerol) containing 0 (control), 2.5, 5, 7.5 or 10 mM GSH. The samples were cooled to 4 C and then frozen in liquid nitrogen vapor. Motility parameters of the sperm were evaluated at 0, 1, 2, 3, 4, 12 and 24 h after thawing. Sperm motility was higher in the 5 mM GSH group than in the control or 2.5 and 10 mM GSH groups; this effect was observed at 1 to 24 h after thawing (P < 0.05). The 5 mM GSH group had a higher sperm viability index at 12 and 24 h after thawing compared with the other groups (P < 0.05). Acrosome integrity, evaluated at 4 h after thawing, was greater in two of the GSH-treated groups (5 and 10 mM) compared with the control. Lipid peroxidation (LP) levels immediately after thawing were lower in the 5 and 10 mM GSH groups compared with the control, while those at 12 h after thawing did not differ significantly. Frozen-thawed semen in the 5 mM GSH group was used for transcervical insemination of 4 bitches, resulting in delivery of 5 puppies from 2 bitches. These results indicate that supplementation of semen extender with 5 mM GSH was effective in improving motility, longevity and acrosomal integrity and inhibiting LP levels in post-thaw canine spermatozoa, without any adverse impacts on full-term development after transcervical insemination.     

Bacillus cereus from the environment is genetically related to the highly pathogenic B. cereus in Zambia

To follow-up anthrax in Zambia since the outbreak in 2011, we have collected samples from the environment and the carcasses of anthrax-suspected animals, and have tried to isolate Bacillus anthracis. In the process of identification of B. anthracis, we collected two isolates, of which colonies were similar to B. anthracis; however, from the results of identification using the molecular-based methods, two isolates were genetically related to the highly pathogenic B. cereus, of which clinical manifestation is severe and fatal (e.g., pneumonia). In this study, we showed the existence of bacteria suspected to be highly pathogenic B. cereus in Zambia, indicating the possibility of an outbreak caused by highly pathogenic B. cereus.     

Effects of chronic mild food restriction on behavior and the hypothalamic malonyl‐CoA signaling pathway

Depression induces anorexia, leading to suppressed feeding behaviors and energy intake. Previously, we revealed that chronic social defeat induced a mild suppression of feeding in rats with elevated levels of hypothalamic malonyl‐CoA which regulates feeding. Therefore, we attempted to elucidate the effects of chronic mild food restriction on behavior and on hypothalamic malonyl‐CoA. The chronic mild food restricted rats were fed a restricted diet approximately 80% to 90% amount of diet compared to the control for 5 weeks. Ratios of restriction were adjusted with feed consumption in the chronic social defeat stressed rats. Chronic mild food restricted rats exhibited a suppression of body weight gain similar to that of the chronic social defeat stressed rats. Also these rats showed increased time spent in the center area of an open field (OF), prolonged immobility time in forced swim, increased phosphorylation of hypothalamic adenosine monophosphate‐activated protein kinase (AMPK) and acetyl‐CoA carboxylase and a decreased concentration of hypothalamic malonyl‐CoA. Weight of the adrenal glands, locomotion in an OF, mitogen‐activated protein kinase cascade and calcium/calmodulin‐dependent protein kinases II in the hippocampus were not affected by chronic mild food restriction. Our findings suggest that chronic mild food restriction activates AMPK following a decreased hypothalamic malonyl‐CoA.     

臂形草内生菌特异DNA片段的克隆及其分子鉴定

进行了臂形草内生菌特异DNA片段的克隆及分子鉴定方法的研究。从5个臂形草品种分离的5个内生菌纯培养特中，提取基因组DNA，通过140条随机引物的RAPD分析，引物OPAK10扩增出1条约500bp的共有PCR谱带。此DNA片段命名为BE1。对BE1进行分离、纯化，经点杂交证实BE1为臂形草内生菌特异DNA片段。进一步将BE1片段进行回收、克隆和DNA序列分析。BE1在基因库中进行序列分析，未发现相关同源片段。臂形草内生菌特异DNA片段的获得，为建立一种准确、快速检测臂形草内生菌及特异内生菌方法奠定了分子生物学基础。     

Stereochemistry and biosynthesis of (+)-lyoniresinol,a syringyl tetrahydronaphthalene lignan in <Emphasis Type="Italic">Lyonia ovalifolia</Emphasis> var. <Emphasis Type="Italic">elliptica</Emphasis> II: feeding experiments with <Superscript>14</Superscript>C labeled precursors

To clarify the biosynthetic pathway for syringyl lignans, especially syringyl tetrahydronaphthalene lignans and formation of the C2–C7′ linkage, production of (+)-lyoniresinol (LYR) and its predicted intermediates [syringaresinol (SYR), 5,5′-dimethoxylariciresinol (DMLR), and 5,5′-dimethoxysecoisolariciresinol (DMSLR)] in Lyonia ovalifolia var. elliptica was investigated by means of feeding experiments with radiolabeled precursors. Following individual administration of l-[U-¹⁴C]phenylalanine (Phe), [8-¹⁴C]sinapyl alcohol (SA), and [8,8′-¹⁴C]SYR to excised young shoots of L. ovalifolia and their subsequent metabolism, free [¹⁴C]lignans and [¹⁴C]lignan glycosides were extracted with methanol from stems and leaves and were divided into ethyl acetate-soluble fractions (lignans) and aqueous fractions (lignan glycosides), respectively. Using a combination of xylanase, cellulase, and β-glucosidase, the glycosides were hydrolyzed to liberate [¹⁴C]lignans as aglycones. l-[U-¹⁴C]Phe was incorporated into (+)-[¹⁴C]SYR [stem 0.38%, 8% enantiomeric excess (e.e.)], (−)-[¹⁴C]SYR (leaves 2.75%, 72% e.e.), (+)-[¹⁴C]DMLR (stem 0.07%, 18% e.e. and leaves 0.009%, 58% e.e.), (−)-[¹⁴C]DMSLR (stem 0.03%, 46% e.e. and leaves 0.05%, 20% e.e.), (+)-[¹⁴C]LYR (leaves 0.013%, 22% e.e.) and glycosides of (+)-[¹⁴C]LYR (stem 0.036%, 50% e.e.) in 24h. Based on the percent incorporation and enantiomeric composition of the lignans, the biosynthetic pathway of (8R,8′R)-(+)-LYR was proposed as follows: a nonselective dehydrogenative dimerization of sinapyl alcohol yields (±)-SYR, which is reduced with low specificity to give (8R,8′R)-(+)-DMLR. This is cyclized to directly give (+)-LYR as well as reduced again to (8R,8′R)-(−)-DMSLR. Although further transformation of (−)-DMSLR also leads to the formation of (+)-LYR, cyclization could be a main pathway for (+)-LYR biosynthesis. This report was presented at the IAWPS 2005 International Symposium on Wood Science and Technology, Yokohama, November 2005