Changes in the microbial community structure in soils treated with a mixture of glucose and peptone with reference to the respiratory quinone profile

Based on the respiratory quinone profile, changes in the structure of microbial communities in the soil samples from Nagoya University Farm were monitored after the treatment with 1% of a mixture of glucose and peptone. Samples of two soils differing in the fertilization history were examined: CF-soil with the application of only chemical fertilizers and FYM-soil with the application of only farmyard manure at a high rate. In the CF-soil, the amount of water-soluble organic carbon (WOC), indicator of the mixture of glucose and peptone, decreased to the original level after 14 d. After 7 d, the soil pH reached the maximum level, then decreased gradually. Changes in the inorganic nitrogen levels in the water extract also reflected the 14-d period of mineralization. The amount of respiratory quinones reached maximum levels after 7 d and gradually decreased, reflecting the changes in the microbial biomass. The quinone composition significantly changed during the 14-d period and returned to a profile similar to the original one after 28 d. Diversity of quinones significantly decreased during the 14-d period due to the predominance of ubiquinone with 9 isoprenoid units. In the FYM-soil, the amount of WOC decreased to the original level after 1 d, and the pH and inorganic nitrogen levels in the water extract reflected the one-day mineralization period, and nitrification started after 3 d. Although the amount of quinones indicated an increase in the microbial biomass for 14 d, the quinone composition did not change. These findings suggested that long-term application of farmyard manure resulted in stable microbial communities in response to the incorporation of organic matter in soil.     

Improvement of preservation method for ammonia-oxidizing bacteria by freeze-drying

Long-term preservation of ammonia-oxidizing bacteria had been achieved until now by using the silica gel-freezing method. Recently, the freeze-drying method for the preservation of microorganisms has been widely used, and has been applied to the preservation of ammonia-oxidizing bacteria. The 10% skim milk-l% pyruvate or glutamate ratio was optimum as a dispersion-medium, and the preservation temperature of the freeze-dried cells was effective at -80°C. This method of preservation was found to be considerably more effective than the silica gel-freezing method.     

Lysozyme-glucose stearic acid monoester conjugate formed through the Maillard reaction as an antibacterial emulsifier

Lysozyme-glucose stearic acid monoester (HEL-GE) conjugate was prepared through the Maillard reaction as an antibacterial emulsifier. The molar ratio of GE to HEL was 1:1. The isoelectric point was 6-7, which is lower than that of native HEL. Spectroscopic studies indicated that the alpha-helix content was slightly lower but the conformation around Trp had not changed and that the surface of the conjugate was covered with the GE moiety. The conjugate maintained approximately 53-57% of the enzymatic activity of native HEL at 40-60 degrees C and exhibited considerable resistance to proteolysis. The denaturation temperature of the conjugate was approximately 74 degrees C, somewhat higher than that of control HEL, whereas the enthalpy was about one-third of that of control HEL. The emulsifying activity of the conjugate and the emulsion stability were much enhanced compared to those of native HEL, and the conjugate maintained approximately 70% of the bactericidal activity.     

Functional changes in beta-lactoglobulin by conjugation with cationic saccharides

Bovine beta-lactoglobulin (beta-LG) was conjugated to each of three cationic saccharides [glucosamine (GlcN), chitopentaose (CPO), and chitosan (CHS)] by means of a water-soluble carbodiimide or by the Maillard reaction in an effort to improve the functional properties of beta-LG. The molar ratios of beta-LG to the cationic saccharide in the beta-LG-GlcN, beta-LG-CPO, and beta-LG-CHS conjugates were 2:1, 2:5, and 2:1, respectively. Fluorescence studies indicated that the conformation around Trp had changed in each conjugate and that the surface of each of the conjugates was covered with a saccharide chain. Structural analysis using monoclonal antibodies indicated that the conformation around (15)Val-(29)Ile (beta-sheet region) in beta-LG-GlcN and beta-LG-CPO had changed but that in beta-LG-CHS was maintained, whereas the conformation around (125)Thr-(135)Lys (alpha-helix region) in the conjugates had changed. The emulsifying activity of beta-LG was improved by conjugation with CPO or CHS, and aggregation of beta-LG was suppressed by conjugation with CHS. Reduction of the antigenicity and immunogenicity of beta-LG was achieved by conjugation with CHS.     

Quantitative assessment of muscle mass and gene expression analysis in dogs with glucocorticoid-induced muscle atrophy

The present study aimed to quantitatively evaluate muscle mass and gene expression in dogs with glucocorticoid-induced muscle atrophy. Five healthy beagles received oral prednisolone for 4 weeks (1 mg/kg/day), and muscle mass was then evaluated via computed tomography. Histological and gene expression analyses were performed using biopsy samples from the biceps femoris before and after prednisolone administration. The cross-sectional area of the third lumbar paraspinal and mid-femoral muscles significantly decreased after glucocorticoid administration (from 27.5 ± 1.9 to 22.6 ± 2.0 cm² and from 55.1 ± 4.7 to 50.7 ± 4.1 cm², respectively; P<0.01). The fast- and slow-twitch muscle fibers were both atrophied (from 2,779 ± 369 to 1,581 ± 207 μm² and from 2,871 ± 211 to 1,971 ± 169 μm², respectively; P<0.05). The expression of the growth factor receptor-bound protein 10 (GRB10) significantly increased after prednisolone administration (P<0.05). Because GRB10 suppresses insulin signaling and the subsequent mammalian target of rapamycin complex 1 activity, increased expression of GRB10 may have resulted in a decrease in protein anabolism. Taken together, 1 mg/kg/day oral prednisolone for 4 weeks induced significant muscle atrophy in dogs, and GRB10 might participate in the pathology of glucocorticoid-induced muscle atrophy in canines.     

Quantitative structure-activity analysis of larvicidal 1-(substituted benzoyl)-2-benzoyl-1-tert-butylhydrazines against Chilo suppressalis

The larvicidal activity of a number of 1-(substituted benzoyl)-2-benzoyl–1 -ten-butylhydrazines against the rice stem borer (Chilo suppressalis Walk.) was measured. Variations in the activity were examined quantitatively using physico-chemical substituent and molecular parameters and regression analysis. The results indicated that the molecular hydrophobicity and the electron-withdrawing inductive/ field effect of ontho substituents are favourable to larvicidal activity. The bulkiness of substituents at the meta and para positions was unfavourable to activity, substitution at the para position being more unfavourable than that at the meta position in terms of van der Waals' volume. The 2,3–, 2,5- and 2,6-disubstitution patterns were also unfavourable to activity. Reductions in larvicidal activity caused by the 2,6-,- 2,3,5- and 2,3,4,5-substitutions were greater than those induced by the 2,3- and 2,5-disubstitutions. When the sum of contributions from favourable effects is greater than that from unfavourable effects, the larvicidal activity is expected to be superior to that of the unsubstituted compound.     

Frequency of and risk factors for epistaxis associated with exercise-induced pulmonary hemorrhage in horses: 251,609 race starts (1992-1997)

OBJECTIVE: To determine the frequency of epistaxis during or after racing among racehorses and identify factors associated with development of epistaxis. DESIGN: Retrospective study. SAMPLE POPULATION: 247,564 Thoroughbred and 4,045 Anglo-Arab race starts. PROCEDURE: Race start information (breed, age, sex, racing distance, and race type) was obtained for Thoroughbred and Anglo-Arab horses racing in Japan Racing Association-sanctioned races between 1992 and 1997. All horses that raced were examined by a veterinarian within 30 minutes of the conclusion of the race; any horse that had blood at the nostrils was examined with an endoscope. If blood was observed in the trachea, epistaxis related to exercise-induced pulmonary hemorrhage (EIPH) was diagnosed. RESULTS: Epistaxis related to EIPH was identified following 369 race starts (0.15%). Frequency of EIPH-related epistaxis was significantly associated with race type, age, distance, and sex. Epistaxis was more common following steeplechase races than following flat races, in older horses than in horses that were 2 years old, following races < or =1,600 m long than following races between 1,601 and 2,000 m long, and in females than in sexually intact males. For horses that had an episode of epistaxis, the recurrence rate was 4.64%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that frequency of EIPH-related epistaxis in racehorses is associated with the horse's age and sex, the type of race, and the distance raced. The higher frequency in shorter races suggests that higher intensity exercise of shorter duration may increase the probability of EIPH.     

Immunohistochemical detection of apolipoprotein B-100 and immunoglobulins (IgA, IgM, IgG) in the splenic arteries of aging dogs

Accumulation of lipids and hyalinosis in the splenic arteries of aged dogs are frequently detected by routine histopathologic examinations. The purpose of this study was to pinpoint the localization of canine apolipoprotein B-100 (CApoB-100) and immunoglobulins (IgA, IgM, IgG) in the splenic arteries of aging dogs (n = 80) through the use of immunohistochemical techniques. CApoB-100 deposits were found in the subendothelial space, extracellular matrix, and atheromatous lesions in the tunica media of the arteries in dogs > or = 6 years of age. Foamy cytoplasm of the infiltrated macrophages was also CApoB-100 immunopositive. In dogs > or = 10 years of age, almost all central arteries were CApoB-100 immunopositive. Hyaline deposits within the wall were characterized by immunopositivity against canine IgA, IgM, IgG, and albumin. Lipid accumulation in splenic arteries may be an age-related lesion and a precursor of the atheromatous plaques associated with splenic hemorrhage and infarcts later in life. In addition, deposition of immunoglobulins, probably mediated by immune complexes, may play an important role in the development of canine vascular diseases similar to human disease.     

Changes in serum thyroid hormone levels in newborn calves as a diagnostic index of endemic goiter

Maximum serum thyroxine (T4) and triiodothyronine (T3) levels of healthy calves were seen at 1 day after birth, and thereafter rapidly decreased until 5 days after birth. They stabilized until 2 weeks after birth, then gradually decreased until 4 weeks after birth. Serum T4 levels of calves with endemic goiter tended to be lower than those of healthy ones, but showed similar levels to those of adult cows. T3 levels of calves with goiter were similar to those of healthy ones, but showed higher variation. T4/T3 ratio of calves with goiter were significantly lower than those of healthy ones and adult cows. While individual levels of serum T4 and T3 at just after birth could not be considered as a diagnostic index, the T4/T3 ratio could be adopted as a diagnostic index of endemic goiter.     

Differences between carcass grades in some morphological, behavioral and physiological measurements at early and middle stages of fattening in crossbred steers

The differential variables for carcass grades were identified from morphological, behavioral and physiological measurements of young steers at a commercial farm. Thirty‐five Japanese Black × Holstein steers aged 6–10 months were randomly assigned to three pens. The steers had free access to ryegrass hay and a restricted amount of concentrate diet for the first 6 months of trial. They were then given oat straw ad libitum and an additional concentrate diet until slaughter. Behavioral observations (15 categories) were carried out once every 2 weeks for 2 h after feeding. Bodyweight was measured, blood sampled (for seven metabolic hormones and five metabolites), ultrasonic scanning conducted and physical measurements taken (10 parts for each animal) in months 1, 3 and 5 (early fattening stage, EFS) and in months 7, 9 and 11 (middle fattening stage, MFS) of the trial. Temperament scores during these procedures, entry order into restraint stalls, social rank and average daily gain were also assessed. A factor analysis and one‐way anova were used to identify clusters of variables which had different factor scores between carcass grades. As for the carcass yields, a cluster of chest girth and depth, bodyweight, withers and hip height, and rump length (P < 0.01), and a cluster of the frequency of scratching the body with facilities (P < 0.05) were identified as differential variables in the EFS. In the MFS, a cluster of concentrations of cortisol and epinephrine, and the frequency of eating straw were identified (P < 0.10). As for the carcass quality grade, a cluster of social rank, triglyceride concentration and the frequencies of stand‐chewing cud and eating hay, and a cluster of the frequency of investigating facilities (both P < 0.10) were identified in the EFS. In the MFS, a cluster of hip height, bodyweight, cannon circumference, chest depth, withers height and chest girth, and a cluster of the frequencies of eating a concentrate diet and stand‐chewing the cud, and social rank (both P < 0.10) were identified. In conclusion, bodyweight, chest girth and depth, withers and hip height were identified as good differential variables for future carcass grades of young steers. Facility enrichment that encourages steers to eat hay in the EFS, and to eat a concentrate diet in and after the MFS would be effective in upgrading carcass grades. High incidence of investigating and scratching the body with facilities and stand‐chewing the cud might lead to lower carcass grades.