Storage of maitake mushroom (Grifola frondosa) culture medium after harvesting fruit bodies is an effective pretreatment for ethanol conversion

The storage of spent maitake culture medium (SMCM) under various conditions was investigated as a potential pretreatment of SMCM for increased ethanol conversion. When SMCM was stored at 25°C for 12?weeks, the glucose yield by enzymatic saccharification increased from 22 to 52%. Selective degradation of lignin and hemicellulose occurred in SMCM during storage. The optimal storage temperature of SMCM was at the active growing temperature of maitake mycelium, which is between 25 and 30°C. Storage of SMCM under anaerobic conditions did not increase the glucose yield compared to non-stored conditions. The glucose yield from the SMCM stored for 4?weeks at 25°C was increased by about 30% with either NaOH or vibrating ball milling pretreatment. After 12?weeks of storage, the glucose yield from SMCM without any other pretreatment was higher than that of non-stored SMCM with additional pretreatments. An ethanol yield of 42.1% was obtained from SMCM stored for 12?weeks by simultaneous saccharification and fermentation, which was comparable to yields after NaOH (41.3%) or vibrating ball milling (44.1%) pretreatments. Therefore, storage of SMCM is a very useful pretreatment for bioethanol production.     

Antimicrobial susceptibility of Escherichia coli isolated from feces of wild cranes migrating to Kagoshima, Japan

Susceptibility to 13 antimicrobial agents was examined for 138 Escherichia coli isolates obtained from 192 fecal samples of wild cranes that migrated for wintering to the Izumi plain, Kagoshima prefecture in Japan. The numbers of isolates that were resistant to the antimicrobials used in this study are as follows: oxytetracycline (OTC), 22 isolates; minocycline, 7 isolates; ampicillin (ABPC), 4 isolates; nalidixic acid, 4 isolates; enrofloxacin, 2 isolates; kanamycin, one isolate. Multidrug resistant isolates exhibiting 2-4 drug resistances were obtained. All of the OTC-resistant isolates carried either the tet (A) or tet(B) gene. The bla(TEM) gene was found in all of the ABPC-resistant isolates.     

CONTRAST AGENT Gd‐EOB‐DTPA (EOB·Primovist®) FOR LOW‐FIELD MAGNETIC RESONANCE IMAGING OF CANINE FOCAL LIVER LESIONS

Contrast‐enhanced magnetic resonance (MR) imaging with a new liver‐specific contrast agent gadolinium‐ethoxybenzyl‐diethylenetriamine penta‐acetic acid (Gd‐EOB‐DTPA; EOB·Primovist®) was studied in 14 normal beagles and 9 dogs with focal liver lesions. Gd‐EOB‐DTPA accumulates in normally functioning hepatocytes 20 min after injection. As with Gd‐DTPA, it is also possible to perform a dynamic multiphasic examination of the liver with Gd‐EOB‐DTPA, including an arterial phase and a portal venous phase. First, a reliable protocol was developed and the appropriate timings for the dynamic study and the parenchymal phase in normal dogs using Gd‐EOB‐DTPA were determined. Second, the patterns of these images were evaluated in patient dogs with hepatic masses. The optimal time of arterial imaging was from 15 s after injection, and the optimal time for portal venous imaging was from 40 s after injection. Meanwhile, the optimal time to observe changes during the hepatobiliary phase was from 20 min after injection. In patient dogs, 11 lesions were diagnosed as malignant tumors; all were hypointense to the surrounding normal liver parenchyma during the hepatobiliary phase. Even with a low‐field MR imaging unit, the sequences afforded images adequate to visualize the liver parenchyma and to detect tumors within an appropriate scan time. Contrast‐enhanced MR imaging with Gd‐EOB‐DTPA provides good demarcation on low‐field MR imaging for diagnosing canine focal liver lesions.     

In vitro susceptibility and a new point mutation associated with tylosin-resistance in Japanese canine intestinal spirochetes

The in vitro susceptibilities of six commonly used antimicrobial agents against 29 isolates of intestinal spirochetes isolated from dogs in Japan were examined by the agar dilution technique. In addition, the genetic basis of tylosin resistance in in vitro selected resistant mutants of two reference strains and three tylosin-susceptible field isolates obtained by three successive subcultures on blood agar containing 1 microg/ml of tylosin was investigated. Carbadox was the most active (MIC: < 0.00625) of all the antimicrobial agents. Although all the isolates were susceptible to tylosin, some were resistant to erythromycin. Tiamulin, lincomycin and dimetridazole were also very active against the isolates. All the resistant isolates did not harbor any plasmids. In vitro selected tylosin-resistant mutants of previously tylosin-susceptible isolates showed a new mutation in which their adenine at the base position equivalent to 2062 of 23S rDNA of Escherichia coli has been replaced by cytosine. These findings may both provide guidance towards the proper choice of antimicrobial agents for the treatment of canine intestinal spirochetosis, and add to the understanding of the genetic basis of tylosin resistance.     

Seroepidemiologic studies on Babesia caballi and Babesia equi infections in Japan

Antibodies to Babesia caballi and Babesia equi were examined on a total of 2,019 horse serum samples that had been collected in 1971-1973 by the National Institute of Animal Health by enzyme-linked immunosorbent assay (ELISA) using recombinant proteins and by Western-blot analysis. Based on the criterion for positivity by ELISA, 5.4% (109/2,019) and 2.2% (44/2,019) had antibodies against B. caballi and B. equi, respectively. The ELISA-positive sera were further examined by Western blot; 30/109 for B. caballi and 2/ 44 for B. equi were positive for native B. caballi or B. equi, but none of them was seropositive for both infections. Based on the results of this study, further investigations should be required to survey horses that have arrived in Japan relatively recently and tick vectors of equine Babesia using ELISA with some recombinant protein, a parasite detection method in an in vitro culture of equine Babesia, and PCR testing.     

Phenotype and function of murine peritoneal cavity macrophage derived-dendritic cells

The accessory activity was reported in murine peritoneal cavity macrophage derived dendritic cells (PEC-DC) in a mixed lymphocyte reaction (MLR). Here we continue the characterization of the generated PEC-DC using the criteria of morphology, phenotype and other accessory function. We have demonstrated that murine peritoneal cavity macrophages can be induced to differentiate in vitro into cells exhibiting typical dendritic cell (DC) morphology, phenotype and function. The proliferative capacity of the progenitors was amplified in the first step of the culture (day 0-7) using a combination of early cytokines: interleukin 4 and granulocyte-macrophage colony-stimulating factor. The second step of the culture started at day 7 with the removal of early growth factors to allow differentiation and final maturation of DC during 2 days of culture with interferon gamma plus either Toxoplasma lysate antigen (TLA) or lipopolysaccharide (LPS), a bacterial agent as a DC maturing agent. The resulting DC population exhibited typical DC morphology and expressed higher levels of MHC class II and the co-stimulatory molecules CD80 and CD86 compared to the untreated peritoneal cells. The generated DC cells efficiently presented soluble protein antigen to CD3(+) spleen T cells.     

Soluble organic nitrogen pools in greenhouse and open field horticultural soils under organic and conventional management: A case study

The aim of this study was to investigate the influence of four different horticultural management practices in open field and in greenhouse conditions under organic and conventional cultivation on the amount of soluble organic nitrogen (SON) present in the soil. Soils used in greenhouses and open field cultivation were sampled in Shanghai, China, where organic farming has been conducted for 3 years or conventional faming has been continued in the same area. The amounts of SON, nitrate (NO₃^?) and ammonium (NH₄⁺) were greater in the greenhouse soils than those under open field cultivation, which indicated a higher degree of soil management was imposed under greenhouse conditions. Greenhouse cultivation is also known to accelerate the turnover of SON in the soils, which may explain the significantly higher amounts of SON present in these soils. Organic farming, which does not use artificial fertilizers and pesticides, also resulted in significantly higher amounts of SON (average 42.10 mg kg^?1) compared with soils under conventional faming (24.59 mg kg^?1). The reasons for the observed differences in pool sizes of soluble inorganic nitrogen (SIN) and NO₃^? in the greenhouse soils and the open fields include (a) the heavy application of both complex fertilizer and organic fertilizer that exceeded crop requirements and (b) warmer temperatures and moist soils in the greenhouses, which are likely to lead to greater rates of N cycling compared with the open field soils. These results suggest that SON may be an important source of N in all horticultural systems, representing a pool of labile N readily available for plant growth. However, its concentration is less sensitive to different management practices than SIN. In contrast to SON, the total soluble nitrogen and inorganic N (SIN) pools varied widely with the different management practices although they were dominated by NO₃^? in all treatments. Soil organic N was positively related to dissolved organic carbon and NO₃^? contents. This relationship indicates that NO₃^? and dissolved organic matter play a key role in the retention of SON in soil.     

Molecular characterization of Theileria orientalis piroplasm protein encoded by an open reading frame (To ORF2) in a genomic fragment

In the present study, a novel antigenic protein expressed in the piroplasm stage of Theileria orientalis was characterized. A 4,707 bp genomic fragment amplified by PCR contained two open reading frames (ORFs). The deduced amino acid sequence of the first ORF showed significantly high similarlity to the ubiquitin carboxy terminal hydrolases/proteases while the second ORF (To ORF2) showed homology to several surface antigens of plasmodia. To ORF2 was expressed to determine whether the protein product is expressed by the parasite. In western blot analysis, bovine antiserum from a T. orientalis-infected calf recognized the recombinant protein containing a C-terminal part of the ORF expressed by baculovirus system. Western blot analysis with the anti-To ORF2 mouse serum recognized a 48 kDa protein in T. orientalis piroplasm lysates. Indirect immunofluorescence antibody test by confocal scanning laser microscopic analysis showed that antisera against the recombinant protein recognized T. orientalis piroplasm in the infected erythrocyte. The results from this study indicate that To ORF2 protein is expressed at the piroplasm stage and is immunogenic. This novel antigenic To ORF2 protein could be exploited for vaccine development against bovine piroplasmosis.