Evaluation of truncated LipL32 expressed by Escherichia coli and Pichia pastoris for serodiagnosis of Leptospira infection in rodents

The applicability of the recombinant LipL32 for serodiagnosis of leptospiral infection in field rodents was assessed in this study. An immunodominant region of LipL32 was determined by monoclonal antibodies, and then, truncated LipL32 (tLipL32) was designed to contain the region (87–188th amino acid). The tLipL32 was compared between two recombinant expression hosts Escherichia coli and Pichia pastoris in ELISA. With field rat sera, tLipL32 expressed by P. pastoris (tLipL32p) had high antigenicity without background reactions, while tLipL32 expressed by E. coli (tLipL32e) showed high background reactions, which were reduced by pre-adsorption of sera with E. coli. To evaluate tLipL32-ELISA, field rat sera were tentatively divided into a Leptospira infection positive (12 sera) and a negative group (12 sera) based on the results from flaB gene PCR of kidney samples and WB with whole Leptospira cell. Consequently, the sensitivity of tLipL32p-ELISA for field rat sera was 83% . A similar result was obtained from tLipL32e-ELISA with adsorbed sera, (92%). However, sensitivity of tLipL32e-ELISA using sera without an adsorption treatment was 50%. Regardless of the expression host, tLipL32-ELISA had 100% specificity and sensitivity in experimentally infected laboratory rats. These results suggest that recombinant LipL32 expressed by P. pastoris is more applicable for serodiagnosis in field rats due to a lack of background reaction.     

Susceptibility of Commercial Tomato Cultivars to Bacterial Wilt in Hydroponic System

Commercially available tomato cultivars were hydroponically cultured for inoculation, with Ralstonia solanacearum (K-101), which causes bacterial wilt, by pouring an inoculum suspension into the nutrient solution. Cultivar susceptibility to the bacteria was evaluated, based on the highest percentage of wilting. Because the length of time for wilt appearance varied among cultivars, some cultivars appeared to be suppressive to the translocation and/or multiplication of the invading pathogen. Thus, this hydroponic inoculation system is effective for examining levels of susceptibility in tomato cultivars to bacterial wilt. Received 13 December 2000/ Accepted in revised form 27 March 2001     

Three dogs under 2 years of age with heartworm caval syndrome

Heartworm caval syndrome (CS) occurred in three dogs under 2 years of age. A 23-month-old dog was recovered by surgical and medical treatments, but the other 2 dogs (15 and 21 months old) died. Necropsy demonstrated 12 heartworms in the 15-month-old dog and 8 worms in the 21-month-old dog. Histopathologically, pulmonary arterial embolism caused by dead worms and thrombi were observed in these cases. The findings suggested that CS could develop regardless of canine age and worm burden if pulmonary arterial embolism related to worm death or thrombus formation were induced.     

Comparison of homogeneity and heterogeneity of residual variance using random regression test-day models for first lactation Japanese Holstein cows

Using a large‐scale data set that included first lactation test day records from 1975 to 2000 for Japanese Holsteins, genetic parameters for milk yield were estimated by using random regression (RR) test‐day models (TDM) with heterogeneous and homogeneous residual variances. It is necessary for the RR‐TDM to include a function that explains the shape of the lactation curve. The RR‐TDM with the LW curve, which combined Wilmink's curve and a Legendre polynomial, was used for fitting the model for milk yield. In recent years, increases in residual variance have been noted for Japanese dairy cattle. Thus, three kinds of heterogeneous residual variance over the calving year were considered: H1, H2 and HG. Linear and quadratic exponential functions for the calving year were used in H1 and H2, respectively. Residual variance of HG was divided into five groups according to calving year. Homogeneous residual variance was HO. All heterogeneous residual variances increased with calving year in an almost linear fashion. Residual variance increased over the study period. However, there is no need to consider heterogeneous residual variances in genetic evaluations, because the heterogeneity of residual variance over the years did not affect the ranking of top sires and cows.     

Treatment of cow-waste slurry by a microbial fuel cell and the properties of the treated slurry as a liquid manure

Resource recycling and the proper treatment of animal waste to reduce its environmental impact are currently important issues for the livestock industry. A microbial fuel cell (MFC), a new type of bioreactor, is expected to play roles in both waste‐water purification and energy recovery. However, the generation of electricity from cow waste has not yet been examined. In this study, using an MFC, we examined the possibility of generating electricity from dairy‐cow waste slurry, and analyzed the properties of the treated slurry as liquid manure for resource recycling. The MFC treatment of the slurry generated electricity in a dose‐dependent manner, and the maximum power output by the MFC from a 1 g of chemical oxygen demand/L slurry was 0.34 mW/m². After the MFC treatment, 84% of the biological oxygen demand in the slurry was removed and three essential fertilizer elements (nitrogen, phosphorus, and potassium) were retained at 84, 70, and 91% levels, respectively. The amount of ammonia nitrogen in the slurry, as an element of fast‐release fertilizer, was increased by 1.9‐fold. Although the treated slurry displayed properties that made it preferable as liquid manure, further studies to improve the electrical power output by the MFC are required for practical use.     

Abietane-type and labdane-type diterpenoids from the cones of Chamaecyparis obtusa

A novel compound, 8(17),12E,14-labdatrien-19-al (trans-communal), was isolated from ethyl acetate extract of young cones of hinoki (Chamaecyparis obtusa Endl.). The chemical structure of the compound was determined mainly with various nuclear magnetic resonance spectral techniques. Its stereochemistry was determined by derivation to a known compound,trans-communol. In addition to this compound, four known compounds — ferruginol, chamaecydin, 12-hydroxy-6,7-seco-abieta-8,l 1,13-triene-6,7-dial, and frans-communic acid — were isolated. All isolated compounds were subjected to an antifeedant bioassay against the pest insectSpodoptera litura. Results of the bioassay showed that chamaecydin and 12-hydroxy-6,7-seco-abieta-8,11,13-triene-6,7-dial had antifeedant activity.Part of this report was presented at 45th Annual Meeting of the Japan Wood Research Society, Tokyo, April 1995     

Termiticidal activity of wood vinegar, its components and their homologues

The termiticidal activity of wood vinegar, its components, and their homologues have been studied. Three kinds of wood vinegar made from the mixed chips ofCryptomeria japonlca andPseudotsga menziesii (wood vinegar A),Quercus serrata (wood vinegar B), andPinus densiflora (wood vinegar C) exhibited high termiticidal activities againstReticulitermes speratus. Acetic acid, which is the largest content of wood vinegar, exhibited high termiticidal acitivity. The contents of organic fraction of wood vinegars and acetic acid might be responsible for the differences in termiticidal activities among these wood vinegars. The structure and termiticidal activity relations of phenols were studied. Phenol with some substituents revealed higher termiticidal activity than benzene derivatives, which have no hydroxyl group; an ortho substituent of phenol plays an important role in termiticidal activity. It has become apparent that high termiticidal activity cannot be obtained by a phenolic hydroxyl group alone; it can be obtained, however, by some substituents, especially an ortho substituent in addition to a phenolic hydroxyl group. The bulkiness of the substituent at the ortho position participates in termiticidal activity; activity decreases as the size of an ortho substituent increases. It is thought that the interaction at the receptor site of termites is affected by the increased size of the ortho substituent.     

Composition of pyrolyzate from Japanese green tea

Green tea is a beverage that is produced by hot-water extraction of the leaves of tea (Camellia sinensis L.). As the amount of green tea consumption has increased, the importance of using the residue has grown. To address this problem, chemical compositions of two pyrolyzates obtained with an electric kiln, L and G, which were obtained from the tea leaf product and its hot-water extraction residue, respectively, were examined. Both pyrolyzates were alkaline, in contrast to the acidic pHs that are typical of pyrolyzates from wood materials. The major constituents were nitrogen-containing compounds, which accounted for up to about 70% (based on gas chromatography-mass spectrometry peak area). Caffeine was the main compound in both samples, although it might not be a pyrolysis product but actually sublimates from the samples and then condenses back into the pyrolyzates. Other nitrogen-containing compounds were formed mainly from proteins and amino acids. Acids and phenols, which are typical components in pyrolyzates obtained from wood materials and contribute to acidic pH, were rare in both samples. Other neutral compounds are known as compounds in pyrolyzates from wood materials. The pyrolyzates obtained in this experiment were novel and may have new uses.Contents of this report was presented in the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, 2002     

Prolonging hypothermic storage (4 C) of bovine embryos with fish antifreeze protein

Embryos obtained via superovulation are necessary for mammalian artificial reproduction, and viability is a key determinant of success. Nonfreezing storage at 4 C is possible, but currently used storage solutions can maintain embryo viability for only 24–48 h. Here we found that 10 mg/ml antifreeze protein (AFP) dissolved in culture medium 199 with 20% (v/v) fetal bovine serum and 25 mM HEPES could keep bovine embryos alive for 10 days at 4 C. We used a recombinant AFP isolated from the notched-fin eelpout (Zoarces elongatus Kner). Photomicroscopy indicated that the AFP–embryo interaction was enhanced at 37 C. Embryos pre-warmed with the AFP solution at 37 C for 60 min maintained high viability, whereas those that were not pre-warmed could live no longer than 7 days. Thus, short-term storage of bovine embryos was achieved by a combination of AFP-containing medium and controlled pre-warming.