Biomedical subjects
T Mitsuoka
Publications and source records attributed to T Mitsuoka.
Evaluation of the anaerobic method for the analysis of fecal microflora of beagle dogs.
The plate-in-bottle method of Mitsuoka et al. (1969) for the counting of fecal bacteria in beagle dogs was superior to an anaerobic jar method. Comparisons of three nonselective media, such as medium 10 supplemented with 10% cecal extract of dogs (designated as M10C), M10 with 10% fecal extract of dogs (M10F), and M10, by the plate-in-bottle method indicated that the visible bacterial counts for M10C were higher than those for M10 and M10F. The high percentage (18.4%) of numbers of the extremely oxygen-sensitive anaerobes to the fecal total counts by using the plate-in-bottle method with M10C was also observed.
Isolation and characterization of rice-straw degrading clostridia from cattle rumen.
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Impact of the advances in age on the gastrointestinal microflora of beagle dogs.
The gastrointestinal microflora of male beagle dogs in two different age groups; I) less than month 12 of age, and II) more than year 11 of age, was compared. No detectable difference occurred on the microflora of stomach, duodenum, jejunum, and ileum in both dogs. Large bowel (cecum, colon and rectum) microflora in both dogs yielded the different microbial populations. In all regions of large bowel, the levels of bacteroides, eubacteria, peptostreptococci, bifidobacteria, lactobacilli, and staphylococci in the elderly dogs were lower than those in the younger animals, whereas the numbers of Clostridium perfringens and streptococci in the elderly animals were higher than those in the youngers. The high incidence of lecithinase-negative clostridia was observed with advances in age, but not that of spiral shaped rods. The result of this study shows that the advances in age of beagle dogs yield some changes in the microbial population of large bowel in the animals.
Interactive effects of verapamil and sympathetic nerve stimulation on the sinus and AV nodes in the canine hearts.
The interactive effects of verapamil and sympathetic nerve stimulation on the sinus and atrioventricular (AV) nodes were studied in 33 bilaterally stellectomized and vagotomized dogs. Sinus cycle length (SCL) and AV nodal conduction time (AH) at a constant drive cycle length (600 msec) were recorded. Sympathetic nerve stimulation (SNS) was performed for 40 sec by electrical stimulation of the right stellate ganglion. Several doses of verapamil (0.5-2.0 microgram/ml, 1.0 ml) were injected directly into the sinus and AV node arteries at 20 sec of SNS. In addition, SNS was repeated at two different intensities after verapamil administration. Verapamil significantly increased the SCL and the AH in a dose dependent fashion, which was reduced by SNS. The reductions of SCL and AH by the SNS were attenuated in the presence of verapamil when the stimulation intensity was weak, but not when the stimulation intensity was strong. It is concluded that verapamil and sympathetic nerve activity interact antagonistically on the sinus node automaticity and AV nodal conduction.
Effect of dietary fat and fiber on fecal flora, bacterial metabolites, and fecal properties in Japanese volunteers.
The effects of dietary fat and dietary fiber (DF) levels in diet on fecal flora, activities of three fecal enzymes, putrefactive metabolites, fecal mutagenicity and fecal properties were studied in eight healthy volunteers. They were given low fat and low DF diet (LF: fat energy ratio was 13.9%, and DF intake was 9.0 g/day) for 10 days, high fat and low DF diet (HF: fat energy ratio was 52.7%, and DF intake was 7.1 g/day) for 10 days, and high fat and high DF diet (HFF: fat energy ratio was 52.0%, and DF intake was 24.8 g/day) for 10 days. No change of fecal flora at the bacterial group level was observed throughout the experimental period, except that the population of lactobacilli showed a tendency to increase in HF period. Fecal activities of beta-glucuronidase, beta-glucosidase and nitroreductase and some putrefactive products were unchanged between LF and HF, while these values decreased in HFF period. No significant change of fecal properties was observed between LF and HF, while by HFF supplementation fecal weight increased and fecal pH value was lower than that in LF and HF. Excretions of iron, zinc and calcium in feces did not increase by high DF supplementation.
Effect of Konjac mannan on intestinal microbial metabolism in mice bearing human flora and in conventional F344 rats.
The effects of dietary Konjac mannan (KM), a frequent ingredient of traditional Japanese foods, on intestinal microbial metabolism and microflora composition were investigated using two laboratory animal models, namely, conventional F344 rats and C3H/He male mice bearing human flora. Dietary KM led to a significant reduction in faecal beta-glucuronidase, nitroreductase and azoreductase activities, and in the production of phenol and indole in the faeces of conventional F344 rats. In the C3H/He male mice bearing human flora, faecal beta-glucuronidase and nitroreductase activities were significantly reduced by KM ingestion, as were the amounts of the putrefactive products, p-cresol and indole, in the faeces. Slight differences in intestinal microflora composition between control and KM diet groups were noted. The results indicate that, in C3H/He male mice bearing human flora, dietary KM may modify microbial metabolism without causing significant alterations in intestinal microflora composition.
Arrhythmogenic properties of the ventricular myocardium in cardiomyopathic Syrian hamster, BIO 14.6 strain.
STUDY OBJECTIVE: Incidence of spontaneous cardiac arrhythmias and inducibility of ventricular arrhythmias in the cardiomyopathic hamster, BIO 14.6 strain, were examined. DESIGN: A 3 min electrocardiogram (ECG) was recorded weekly to 50 weeks of age. Programmed ventricular stimulation was then performed with single and double ventricular premature stimuli during both sinus rhythm and ventricular pacing, and with brief bursts of ventricular pacing to induce ventricular arrhythmias. EXPERIMENTAL MATERIAL: 19 male cardiomyopathic and 10 age matched normal hamsters from 16 to 50 weeks were used for recording the 3 min ECG; 11 cardiomyopathic and 10 normal hamsters aged 50 to 55 weeks were used for the induction of ventricular arrhythmias. MEASUREMENTS AND MAIN RESULTS: Spontaneous and sporadic ventricular premature contractions were documented in 8.3% of 218 ECGs from the cardiomyopathic hamsters between the ages of 30 and 46 weeks, whereas no ventricular arrhythmia was recorded in normal hamsters throughout the entire follow up period. Non-sustained ventricular tachycardia (NSVT) was induced in 7/11 cardiomyopathic hamsters by single ventricular premature stimulus. Either NSVT or ventricular fibrillation was induced in all cardiomyopathic hamsters with a significantly prolonged intraventricular conduction delay and refractory period. In contrast, neither NSVT nor ventricular fibrillation was induced in normal hamsters. CONCLUSION: This study provides the first evidence of the high arrhythmogenicity of the ventricles in cardiomyopathic hamsters.
Proposal of two subspecies of Fusobacterium necrophorum (Flügge) Moore and Holdeman: Fusobacterium necrophorum subsp. necrophorum subsp. nov., nom. rev. (ex Flügge 1886), and Fusobacterium necrophorum subsp. funduliforme subsp. nov., nom. rev. (ex Hallé 1898).
The biological and biochemical properties, DNA base compositions, and levels of DNA-DNA homology of two biovars of Fusobacterium necrophorum were examined. Some differences were found between the two biovars in biological and biochemical properties. The G + C contents of DNAs from biovar A strains VPI 2891T (T = type strain), NCTC 10576, N167, Fn47, and Fn43, were 32, 30, 29, 28, and 31 mol%, respectively. The G + C contents of DNAs from biovar B strains Fn524T, 606, Fn49, Fn45, and 1260 were 30, 31, 27, 31, and 30 mol%, respectively. Labeled DNA from biovar A strain VPI 2891T exhibited 100 to 80% relatedness to DNAs from biovar A strains and 59 to 51% relatedness to DNAs from biovar B strains. Labeled DNA from biovar B strain Fn524T exhibited 100 to 81% relatedness to DNAs from biovar B strains and 71 to 60% relatedness to DNAs from biovar A strains. Therefore, the names Fusobacterium necrophorum subsp. necrophorum subsp. nov., nom. rev. (ex Flügge 1886), and Fusobacterium necrophorum subsp. funduliforme subsp. nov., nom. rev. (ex Hallé 1898), are proposed for Fusobacterium necrophorum biovars A and B, respectively. The type strain of F. necrophorum subsp. necrophorum is strain VPI 2891 (= JCM 3718 = ATCC 25286), and the type strain of F. necrophorum subsp. funduliforme is strain Fn524 (= JCM 3724).
Quantitative fluorometric assay for rapid enzymatic characterization of Bifidobacterium longum and related bifidobacteria.
The quantitative, semi-automated assay described here is an alternative characterization method allowing for highly sensitive and specific detection of bifidobacterial enzymes. Twenty strains of Bifidobacterium longum, including the type strain ATCC 15707, and type strains of 15 other Bifidobacterium species were enzymatically characterized using 20 4-methylumbelliferyl conjugated substrates. Enzyme activities were determined by directly measuring the intensity of fluorescence derived from 4-methylumbelliferone, a fluorescent metabolic by-product. For this method, a Titertek Fluoroskan II fluorometer was used. Enzymes included glycosidases, an esterase, phosphatase, sulphatase, and neuraminidase. B. longum showed strong activity (greater than 1,000 absolute fluorescence units, afu) for alpha-L-Arabinopyranosidase and alpha-L-Arabinofuranosidase, beta-D-Fucosidase, alpha- and beta-D-Galactosidase, alpha-D-Glucosidase, and alpha-D-Mannosidase. No activity (less than or equal to 50 afu) was observed for beta-D-Cellobiosidase, alpha- and beta-L-Fucosidase, beta-D-Glucuronidase, beta-D-Mannosidase, Neuraminidase and Sulphatase. Enzymatic activity profiles in other bifidobacteria were different according to the species. This assay is simple and rapid (6 hr). Special cultural requirements are unnecessary. Results are objective and quantitative. This assay may be a useful tool for bifidobacterial taxonomy.
Influence of sympathetic nerve activity on ventricular arrhythmogenicity in the dog with chronic hypokalemia.
To examine the influence of sympathetic nerve activity on ventricular arrhythmogenicity in the dog with chronic hypokalemia, an electrophysiologic study was performed before and after bilateral stellectomy (BS) in 10 dogs with chronic hypokalemia (2.8 +/- 0.1 mEq/L), which was created by feeding a low-potassium diet and by administering furosemide over a four-week period, and the results were compared with those obtained from 10 dogs with normokalemia (4.7 +/- 0.3 mEq/L) from being fed an ordinary diet over a four-week period. Before BS the incidence of electrically induced ventricular arrhythmias was higher in the hypokalemic than in the normokalemic dogs. After BS it was decreased considerably in the hypokalemic but not in the normokalemic dogs. Heterogeneity of effective refractory period (delta ERP), which was determined as the difference between the longest and shortest effective refractory periods in three sites of the right and left ventricles, was greater in the hypokalemic than in the normokalemic dogs before BS. The delta ERP decreased slightly in the two groups both before and after BS. There was, however, no significant difference in delta ERP in the two groups both before and after BS. Ventricular fibrillation threshold (VFT) was significantly lower in the hypokalemic dogs than in the normokalemic dogs before BS (p less than 0.005). VFT was elevated in the two groups after BS. Percent increase in VFT after BS was significantly greater in the hypokalemic than in the normokalemic dogs. In conclusion, sympathetic nerve activity may play an important role in the increase in ventricular arrhythmogenicity in the presence of chronic hypokalemia.
Isolation and identification of clostridia from the intestine of laboratory animals.
Quantitative and qualitative examination of faecal clostridia from laboratory mice, rats, hamsters and rabbits was carried out by using selective as well as nonselective culture techniques. The 364 isolates were identified where possible, to species level on the basis of physiological and biochemical characteristics. Most isolates from mice and rabbits were identified, i.e. 79.4% and 76.5%, respectively, while only 25.4% and 28.6% of isolates from rats and hamsters were characterized as described species. The most common species of the 16 identified were: C. coccoides, C. cocleatum and C. innocuum from mice; C. cocleatum, C. cochlearium and C. irregularis from rats; C. cocleatum and C. irregularis from hamsters; and C. aminovalericum and C. irregularis from rabbits. Unidentified Clostridium spp. were isolated from each animal species tested and were tentatively classified into 26 groups. None of these groups was found in more than one animal species in this study.
Effects of sympathetic and parasympathetic stimulation on the induction of atrial flutter in dogs with aseptic pericarditis.
To study the effects of sympathetic stimulation (SS) and vagal stimulation (VS) on the induction of atrial flutter (AF) and its cycle length, aseptic pericarditis was produced surgically in 17 adult mongrel dogs. Programmed atrial stimulation was used to induce AF and to determine the effective refractory periods (ERP) and maximum conduction delay (maxCD) of the atrium. These tests were performed before and during stimulation of the right cervical vagus nerve and the right stellate ganglion. Results showed: (1) AF could be induced in animals with short ERP and large maxCD before SS; (2) ERP was significantly shortened, maxCD was significantly increased by VS; (3) During SS, ERP was slightly shortened, but there were no changes in maxCD and the induction rate of AF; (4) The cycle length of AF was shortened by SS, however, the cycle length of AF was shortened more notably by VS than by SS. From these findings, a shortening of the ERP and an increase in maxCD appear to be related to an increased AF induction rate by VS.
Effect of corn fiber residue supplementation on fecal properties, flora, ammonia, and bacterial enzyme activities in healthy humans.
Effects of corn fiber residue (5 g/day for 10 days) on fecal weight, moisture, pH, fecal flora, ammonia content, and on the activities of beta-glucuronidase and beta-glucosidase were investigated in six healthy subjects. Corn fiber residue was remnant of hemicellulose extraction from corn fiber by calcium hydroxide. Fecal weight showed a tendency to increase, and fecal pH did not change during corn fiber residue supplementation. No remarkable changes in the fecal flora at the bacterial group level were observed. Fecal ammonia content and beta-glucuronidase activity per gram of wet feces decreased slightly but the daily output did not change. Fecal beta-glucosidase activities per gram of wet feces increased significantly (p less than 0.05) and the daily output also tended to increase during corn fiber residue supplementation.
Lactobacillus intestinalis (ex Hemme 1974) sp. nov., nom. rev., isolated from the intestines of mice and rats.
The genetic and phenotypic properties of 10 strains identified as Lactobacillus intestinalis sp. nov. were examined. These strains constitute a distinct species which can be differentiated from all of the previously described homofermentative species in the genus Lactobacillus by their carbohydrate fermentation pattern. The guanine-plus-cytosine contents of their DNAs are 33 to 35 mol%. DNAs from 10 other Lactobacillus species did not exhibit significant levels of relatedness to representative strains of the new species. The name Lactobacillus intestinalis (ex Hemme) sp. nov., nom. rev. is proposed for these isolates, and strain Th4 (= ATCC 49335 = JCM 7548) is the type strain.
Staphylococcus schleiferi subsp. coagulans subsp. nov., isolated from the external auditory meatus of dogs with external ear otitis.
A new subspecies, Staphylococcus schleiferi subsp. coagulans, was isolated from the external auditory meatus of dogs suffering from external ear otitis and is described on the basis of studies of 21 strains. Phenotypic studies showed that these strains are more closely related to Staphylococcus intermedius than to other staphylococci, but DNA hybridization studies indicated that they are closely related to Staphylococcus schleiferi N850274T. On the basis of biochemical distinctiveness (positive test tube coagulase test and different carbohydrate reactions) and the etiological importance (frequent isolation from otitis specimens from dogs) of these strains, we propose to classify them as a subspecies of S. schleiferi. The strains of this new subspecies are coagulase tube test, beta-hemolysin, and heat-stable nuclease positive but clumping factor negative. A simple scheme for the differentiation of S. schleiferi subsp. coagulans from the other coagulase-positive staphylococci is presented. The type strain is GA211 (= JCM 7470).
Improved detection of oral spirochetes with an anaerobic culture method.
A method for routine culturing of oral spirochetes was studied in periodontitis patients and subjects with healthy gingivae. Subgingival plaque bacteria, including oral spirochetes were cultured anaerobically using the steel wool jar method, the glove-box procedure and the plate-in-bottle method. They were also observed directly with a phase-contrast microscope. The number of colonies of recovered spirochetes was significantly higher with the plate-in-bottle method when Medium 10 (M10) or M10 supplemented with 10% rumen fluid was used. In almost all cases, spirochetes were detected with this culture method. The ratios of spirochetes from patient samples, cultured by the plate-in-bottle method, and of other microorganisms, cultured in Eggerth-Gagnon (EG) medium by the steel wool jar method, were almost identical of those observed by phase-contrast microscopy. These results indicate that it can be just as possible to culture certain oral spirochetes as other anaerobic bacteria.
Measurement of endotoxic activity in feces of normal horses.
Two chromogenic Limulus amebocyte lysate systems were evaluated for the measurement of endotoxic activity in feces of normal horses. Fecal extracts had neither non-specific nor inhibitory effect on the test reaction, and the two systems with different specificity showed equivalent results. Endotoxic activities in feces of healthy horses averaged 5.7 +/- 2.8 microns/g wet weight. Correlation between the endotoxin levels and the fecal microbial flora could not be determined.