Hemorrhagic necrotizing splenitis in a slaughter pig infected with Arcanobacterium species

A 6-month-old barrow presented with lethargy, inappetence and dysstasia. At necropsy, multiple coalescing hemorrhagic foci were detected in the margins of the spleen. Gram-positive bacilli were isolated from the spleen, kidney, muscle and liver. Comparative 16S rDNA gene sequencing analysis of the isolates (TO16177) revealed that they would be the same species of unpublished Arcanobacterium species strain HJ57-14E (accession no. gi 18873551) (99.7% similarity based on a comparison of 675 bp). Histologic examination of the splenic tissue sections revealed extensive necrosis and inflammation, and gram-positive bacilli were discernible. Multifocal necrosis was also detected in the liver. Immunohistochemically, the isolates were cross-reacted with polyclonal antibodies against Arcanobacterium pyogenes and Actinomyces naeslundii, and the reaction was strong for the latter. Similar reactions were found in the suppurative lesions of the tonsil, and occasionally in the spleen and lymph nodes. The present results indicate that the unpublished Arcanobacterium species induced multiple organ failure accompanied by acute hemorrhagic necrotizing splenitis in this growing-finishing pig.     

猪隐秘杆菌型出血性坏死性脾炎

六月龄去势公猪表现昏睡、食欲不振、站立困难。尸检发现脾脏边缘呈多重出血灶。从脾脏、肾脏、肌肉和肝脏中分离出革兰氏阳性杆菌,对分离株(TO16177)的16S rDNA基因序列比较分析发现,其可能与未发表过的隐秘杆菌属HJ57-14E菌株(登记号:gi18873551)(比较675bp个碱基,相似性达99.7%)为同一个属。脾脏组织切片的组织学检查发现呈广泛性坏死和炎症,其中革兰氏阳性杆菌显而易见。肝脏中可见多病灶的坏死斑。免疫组织化学检测发现,分离株与抗化脓性隐秘杆菌属(Arcanobacterium pyogenes)和内氏放线菌(Actinomyces naeslundii)的多克隆抗体具有交叉反应,且与后者的交叉反应更强烈。相似的反应也见于扁桃体的化脓灶中分离株,偶尔也见于脾脏和淋巴结中的分离株。本研究结果表明,这种未公开发布的隐秘杆菌属的细菌会引起生长肥育猪多器官功能衰竭,而后继发急性出血性坏死性脾炎。     

Identification and characterization of foliar polyphenolic composition in sweetpotato (Ipomoea batatas L.) genotypes

Trials over two years were conducted using 1389 sweetpotato (Ipomoea batatas L.) genotypes collected from all over the world to characterize the polyphenolic composition in sweetpotato leaves. Wide variation was observed in relation to their total and individual leaf polyphenolic constituents. In all genotypes studied, the total polyphenol contents of sweetpotato leaf ranged from 1.42 to 17.1 g/100 g dry weight. The six different polyphenolic compounds were identified and quantified by NMR, FABMS, and RPHPLC analysis procedures. This is the first report of polyphenolic compositions in sweetpotato leaves. The relative levels of polyphenolic acids in sweetpotato leaves were as follows: 3,5-di-O-caffeoylquinic acid > 4,5-di-O-caffeoylquinic acid > chlorogenic acid (3-O-caffeoylquinic acid) > 3,4-di-O-caffeoylquinic acid > 3,4,5-tri-O-caffeoylquinic acid > caffeic acid. The highest 3,4,5-tri-O-caffeoylquinic acid and 4,5-di-O-caffeoylquinic acid occurred at 221 and 1183.30 mg/100 g dry weight, respectively.     

Morphological modifications among oribatid mites (Acari: Oribatida) in relation to habitat differentiation in mangrove forests

We examined morphological modifications among oribatid species in five microhabitats in mangrove forests in the Ryukyu Islands of Japan. A total of 89 oribatid species were recorded from canopy (leaves and branches), bark of flooded trunks (trunks of 0–50 cm high and knee roots), bark of other trunks higher than 50 cm, and littoral algae in mangrove forests, and the forest-floor soil in an adjacent bank forest. There were no significant differences in the body length, body width, and notogastral length among oribatid species from the five microhabitats. The mean sensillus length of the oribatid species from the forest-floor soil was about twice as long as that from the other microhabitats. Claw morphology was characterized by two attributes: number (monodactyly and tridactyly) and length. In the canopy and trunks, the proportion of tridactyl species was higher than that of monodactyl species. On the other hand, the proportion of tridactyl species in the forest-floor soil accounted for only about 20%, and that in the flooded trunks and littoral algae approximated to zero. The mean claw length was larger in the oribatid species from the flooded trunks and littoral algae than in both monodactyl and tridactyl species from every other microhabitat. Trydactyl species with short claws in the arboreal environments might have been selected by a compromise between grip and mobility for unpredictable environmental changes such as wind and rain. The dominance of monodactyl species with a longer claw in the littoral environments implies a consequence of selection for regular tidal flooding, which requires oribatids to grip tighter on the substrate. The modifications in claw morphology of oribatid mites in mangrove forest might be interpreted as adaptations to a difference in the predictability of the environmental conditions of microhabitats.     

Relationship between antibacterial activity of (+)-catechin derivatives and their interaction with a model membrane

(+)-Catechin derivatives with different alkyl chain lengths were synthesized from (+)-catechin and various straight chain alkylaldehydes in the presence of methyl mercaptan, and their antibacterial activities against Gram-positive bacteria were evaluated. The antibacterial activity increased markedly with elongation of the alkyl chain lengths of the derivatives and reached a maximum at a chain of four to seven carbons. Subsequently, interaction of the (+)-catechin derivatives with a model membrane using liposome was investigated. The derivatives with a chain of three carbons or more were found to have very strong affinity for the membrane. The injury action of the derivatives against the membrane was examined with liposome in which calcein was enclosed as a fluorescent indicator. The leakage was observed in the derivatives with chain lengths of four carbons or more. Particularly the derivatives with chains longer than five carbons are considered to destroy the liposome membrane judging from the degree of the fluorescent leakage. These results implied that the lipophilicity and disrupting ability of the (+)-catechin derivatives to the liposome membrane participate in their antibacterial activity.     

Design of hammerhead ribozymes that cleave murine Sry mRNA in vitro and in vivo

As the first step in investigating the possiblity of applying ribozyme technology to artificial control of the sex ratios at birth in farm animals, where the demand for females exceeds that for males, we designed a hammerhead ribozyme (HHRz) and 2 tRNA(val)-hammerhead ribozyme complexes (tRNARz3 and tRNARz4), and examined their effects upon murine Sry mRNA in vitro and in cells. We demonstrated that HHRz and tRNARz3 could effectively cleave the target Sry mRNA in vitro. For the purpose of experiments in vivo, HHRz was cloned into the highly efficient pUC-CAGGS mammalian expression vector (pCAG/HHRz), and the tRNA ribozyme complexes were cloned into the pol III promoter-driven pPUR-KE vector (pPUR/tRNARz3 and pPUR/tRNARz4); the ribozyme vectors were co-transfected with the target vector (pCAG/Sry). A suppressive action (up to approx. 60%) was confirmed for pCAG/HHRz and pPUR/tRNARz3 upon the transiently expressed exogenously introduced Sry in M15 cultured cells.     

Effects of a collagenolytic cell wall component from Fusobacterium necrophorum subsp. necrophorum on rabbit tissue-culture cells

The effects on rabbit tissue-cultured cells of collagenolytic cell wall component (CCWC) from Fusobacterium necrophorum subsp. necrophorum were investigated. Scanning electron microscopy demonstrated that CCWC damaged the cell surfaces of the rabbit granulocytes and hepatocytes but the effects of the cells differed from each other. Granulocytes appeared smooth and morphologically irregular whereas hepatocytes looked rough and had tiny holes in the cell membranes. Differences in cell viability were observed in MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrazolium, inner salt) assay. The findings suggest that cytotoxic activity in vivo may well contribute to the establishment of an initial injury in visceral tissues, and the action of CCWC could increase the chances of survival for an invading F. necrophorum subsp. necrophorum at the first stages of infection.     

Anthocyanin composition and antioxidant activity of the Crowberry (Empetrum nigrum) and other berries

The anthocyanin composition and antioxidant activity of the crowberry ( Empetrum nigrum) were studied. High-performance liquid chromatography (HPLC) combined with a diode array detector and electrospray ionization mass spectrometry were used for identification and quantification of individual anthocyanins. Freeze-dried crowberry powder was extracted with 80% methanol containing 0.5% acetic acid and subjected to HPLC. Thirteen kinds of anthocyanins were identified. The major anthocyanins were cyanidin-3-galactoside and delphinidin-3-galactoside, at 8.04 and 8.62 mg/g extract, respectively. The HPLC profile of crowberry extract was similar to bilberry and blueberry. The total content of anthocyanins in crowberry was 41.8 mg/g extract, higher than the other nine major berry species (2.5-38.8 mg/g extract). The antioxidant activity was also evaluated using the 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azinobis(3-ethybenzothiazoline-6-sulfonic acid) radical quenching assays and the ferric reducing activity power assy. Crowberry extract exerted the strongest antioxidant activity. In conclusion, individual anthocyanins in crowberry were identified and then quantified in this study. Additionally, crowberry is suggested to be associated with a reduction in the risk of developing chronic diseases because of its strong antioxidant activity.     

Effects of Fiber Additives on the Desiccation Crack Behavior of the Compacted Akaboku Soil as A Material for Landfill Cover Barrier

In the daily and final landfill cover barrier system, the hydraulic properties of compacted soil liners and the strength of soil can be adversely affected by desiccation cracking, resulting in the loss of effectiveness and integrity of the containment system as a barrier. Recently, there is an interest of using fiber additive to overcome the desiccation cracking problem. In this study, the desiccation crack test was conducted to investigate the effect of fiber additive on suppressing desiccation cracks in compacted Akaboku soils. Polypropylene (C₃H₆) fiber was used as an additive material for soil sample. The percentages of fiber used were varied as 0.0%, 0.2%, 0.4%, 0.6%, 0.8%, 1.0% and 1.2% (by dry weight of samples). The soil specimens were compacted under the conditions of maximum dry density and optimum water content. The surficial cracking area was measured to determine the crack intensity factor (CIF) of the soil samples. The desiccation crack test results indicated that the percentage of volume change of the compacted soil specimen decreased with addition of fiber. The change in the soil surface area decreased with increasing in the fiber content (FC), and consequently, the volumetric shrinkage strain decreased. The CIF for the soil without fiber (FC?=?0.0%) were significantly higher than the soil with fiber additive. The CIF of soil at FC?=?0.0% decreased from 2.75% to 0.6% for the soil at FC?=?0.2%. It was also found that the maximum crack depth reaches almost 50% of the thickness of the soil without fiber additive. This study suggests the potential application of the fiber additives to soils as an available method to suppress desiccation cracks encountered in landfill cover barriers.