Protective effect of vaccination with culture supernate of M. hyopneumoniae against experimental infection in pigs

The protective activity of Mycoplasma hyopneumoniae inactivated vaccine prepared from sedimented whole cells and cell-free culture supernates was evaluated experimentally using hysterectomy-produced, colostrum-deprived pigs in which mycoplasmal pneumonia had been induced. The culture supernate vaccine containing less than 10(1) colour-changing units (CCU)/0.2 ml of M. hyopneumoniae significantly (P < 0.05) reduced the percentage of lung lesions compared to controls (3.2 +/- 3.9 vs. 12.2 +/- 2.2%), whereas the sedimented whole cells vaccine containing 10(10) CCU/0.2 ml of organisms provided variable protection (18.7 +/- 16.5 vs. 12.2 +/- 2.2%). Serum from the pigs vaccinated with culture supernate reacted with six protein bands of 97, 89, 65, 46, 42 and 41 kDa by immunoblot analysis. From these results, we conclude that vaccination with culture supernate of M. hyopneumoniae can provide protection against M. hyopneumoniae infection and that these antigens in the culture supernate may be closely related to the reduction of lung lesions.     

Pea extracellular Cu/Zn-superoxide dismutase responsive to signal molecules from a fungal pathogen

We previously reported that the release of O₂ ⁻ from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins. The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss a role for PsCu/Zn-SOD1 in the pea defense response. The nucleotide sequence data of PsCu/Zn-SOD1 reported are available in the DDBJ/EMBL/GenBank databases under accession number AB189165.     

Aloe ring spot,a new disease of aloe caused by <Emphasis Type="Italic">Haematonectria haematococca</Emphasis> (Berk. &; Broome) Samuels &; Nirenberg (anamorph: <Emphasis Type="Italic">Fusarium</Emphasis> sp.)

A ring spot disease of Aloe vera was found on leaves of potted seedlings of Aloe vera in Hachijojima and Chichijima Islands, Tokyo. From tissue of ring spot lesions, a fungus producing Fusarium-type conidia was consistently isolated. After 1 month, reddish perithecia of nectriaceous fungus had formed on the colonies of this isolate on PDA. These nectriaceous and Fusarium fungi were identified as Haematonectria haematococca and Fusarium sp., respectively. From a single ascospore isolation, the former was confirmed to be the teleomorph of the Fusarium sp. Typical ring spot lesions were reproduced by artificial inoculations using single ascospore and single conidium isolates. Inoculations of five species of genus Aloe revealed that they were highly susceptible except for A. arborescens. This is the first report of a disease on Aloe caused by H. haematococca (anamorph: Fusarium sp.) in Japan, and it was named aloe ring spot.     

Restriction endonuclease analysis of canine herpesviruses isolated in Japan

Eighteen canine herpesvirus (CHV) isolates from Japan and two reference strains were compared by restriction endonuclease analysis technique using total DNA extracts from cells infected with the viruses. In order to select the suitable restriction endonucleases for differentiation of CHV isolates, ten enzymes were used and three of them, HindIII, XbaI, and PvuII, were found to be useful for strain differentiation. With these enzymes, CHV isolates from unrelated individuals were readily differentiated from each other. In contrast, all the isolates derived from the same litter were not distinguishable on the basis of restriction cleavage patterns. However, slight mobility shifts were observed among the isolates from the same litter or the same individual. The results showed that this method provides a powerful tool for epidemiological surveys of CHV infection.     

Spectrophotometrical characteristics in the near infrared region in beech (<Emphasis Type="Italic">Fagus crenata</Emphasis>) and pine (<Emphasis Type="Italic">Pinus densiflora</Emphasis>) litters at the various decomposing stages

We examined the relationships between the absorptional characteristics in the near infrared region and the chemical changes of decomposing beech (Fagus crenata) and pine (Pinus densiflora) litters. Spectra as well as the concentrations of chemical substances approached each other and converged with decomposition, although both initial characteristics differed markedly between beech and pine. This indicated that the fundamental chemical structures were almost the same, although their organochemical composition differed. Specific absorption bands for lignin, polysaccharide, and protein were identified at 2,140 and 1,670 nm, 2,270, 1,720, 1,590, and 1,216 nm, and 2,350 nm, respectively. Absorbance at 1,670 nm, peculiar band of aromatics, showed a positive correlation with lignin concentration, which suggested the relative increment of aromatics due to condensed lignin in decomposing litters. Absorbance at 2,140 nm, characterized as the C–H bond in HRC = CHR, showed a negative correlation with lignin concentration, which suggested the decrements of some structures such as side-chains in lignin polymers unrelated to aromatics. Absorbance at 2,270, 1,720, and 1,216 nm, specified to O–H/C–O/C–H bonds in saccharide, might reflect the change of polysaccharide during decomposition because they showed a positive correlation to polysaccharide concentration. In the same way, absorbance at 2,350 nm, identified to the C–H/CH₂ bonds in protein, showed a negative correlation to nitrogen concentration in decomposing litters, which might indicate that the C–H/CH₂ bonds in protein decreased with decomposition due to microbial consumption of carbon in protein. Our findings suggested the possibility that the spectral changes indicate the litter digestibility during decomposition and that also explain the compositional change in decomposing litters.     

Assignment of the 1H and 13C NMR spectra of 2-aminobenzamide-labeled galacto- and arabinooligosaccharides

1,4-Linked β-d-galactooligosaccharides with a degree of polymerization (DP) between 1 and 7 and 1,5-linked α-l-arabinooligosaccharides with a DP between 1 and 8 were labeled at their reducing ends with 2-aminobenzamide (2AB) in the presence of sodium cyanoborohydride. The 2AB-labeled oligosaccharides were shown to be homogeneous using high-performance anion-exchange chromatography (HPAEC) and by electrospray ionization mass spectrometry (ESI-MS). The signals in the ¹H and ¹³C nuclear magnetic resonance (NMR) spectra of the 2AB-labeled oligosaccharides were then assigned using one- and two-dimensional NMR spectroscopy. These NMR data will be useful for the structural analysis of enzymatically synthesized galactan and arabinan side chains derived from rhamnogalacturonan I.     

Characterization of the products resulting from ethylene glycol liquefaction of cellulose

The composition of liquefied cellulose in the presence of ethylene glycol (EG) was studied. The liquefied products were fractionated and analyzed with highperformance liquid chromatography and nuclear magnetic resonance. EG-glucosides were detected as the only saccharides and 2-hydroxyethyl levulinate as the highly decomposed compound derived from cellulose. Quantitative analysis of the EG-glucosides and levulinic acid that comes from the levulinate shows the presence of the following mechanism in the EG-liquefaction of cellulose. First, cellulose is degraded and produces considerable amounts of EG-glucosides during the early stage of liquefaction. Then, when liquefaction is prolonged, the glucosides are decomposed, leading to a large quantity of levulinates.     

Chemical analysis of the product in acid-catalyzed solvolysis of cellulose using polyethylene glycol and ethylene carbonate

Degradation and decomposition of cellulose were studied in an acid-catalyzed solvolysis treatment of biomass using polyethylene glycol (PEG) and ethylene carbonate (EC). The solvolysis reaction was followed by a typical reaction system of wood liquefaction that uses sulfuric acid catalyst at 140° or 150°C at atmospheric pressure. The methods of fractionation and chemical analysis of the degraded cellulose in the solvolyzed product are discussed. The solvolyzed product was separated into several fractions, and they were hydrolyzed to release glucose and levulinic acid to determine the quantity of glucosides and levulinates in the solvolysis product. The data clearly showed that the solvolysis reaction had the same mechanism when using PEG or EC. Degradation of cellulose leads to the formation of glucosides, which then decompose, resulting in a levulinic acid structure, and producing a water-insoluble fraction. The conversion rates of both glucosides and levulinates strongly depend on the reaction conditions of the solvolysis. In particular, EC promotes faster conversion of the reactions. The method discussed here is a chemical analytical technique for characterization of the products of wood liquefaction.