The status of distribution of heat resistant Clostridium perfringens in the feces from healthy persons was investigated by using the anti-sera to Hobbs' type 1 through 13. The resistant organisms were detected in about half of the fecal specimens; The isolated organisms were serologically typable by Hobbs' criteria in many cases. Among 21 fecal specimens yielding the resistant organisms, 7 contained exclusively untypable organisms. Two or more serological types were often found from a single specimen. Out of 18 strains isolated from 14 specimens, 7 were classified as type 13, 4 type 1, 2 type 4 and one each type 2, 3, 5, 9 or 10. The resistant organisms were detected at all the examinations made during the period of 7 months or even longer in some persons. Howeverin, some persons. the types of the isolated organisms often changed from time to time not only in the intermittent but also in the permanent carriers.
There have been a numerous reports regarding the severe reactions after the inoculation of tetanus toxoid, specially after its second one. A part of its causes is considered due to such ingredients proteose or peptone of the original toxin media, of which there have been many reports. We have already reported the formula of P-II-Medium, which is simplified in its preparation and conposition for the production of high level of tetanus toxin in routine use (Table 1), while having such strong allergic substances as peptone and heart infusion in its ingredients. This report is primarily concerned with the production of high potent toxin of the peptone-free medium, eliminated allergic substances of high molecular in P-II-Medium and substituted by allergic-free substances of lower molecular (i.e. casein-acid-hydrolysate and heart infusion dialysate) Following are the ingredients for new medium (CHD): Casein-acid-hydrolysate, heart infusion dialysate, tyrosine, tryptophane, cystine as main nitrogen sources, Vitamins: uracil, Ca-pantothenate, pyridoxine-HCl, thiamine-HCl, nicotinic acid, riboflavine, biotin, Metals: Zn SO47 H2O, MnCl2 4 H2O, Salts: NaCl, Na2HPO4, MgSO4 KH2PO4 other ingredients: Adenine-sulfate or adenine-HCl N-Acetyl-Histidine, glutathione, creatine, (folic acid), glucose and reduced iron. (Table 9). The new medium constantly produces the potent tetanus toxin (20-40Lf/ml). It will contribute to prophylaxis for tetanus particularly for the adult, and in its turn to the public health in general.
Effect of ultraviolet irradiation on the acidfastness of uneasily culturable Mycobacteria, M. paratuberculosis, and unculturable Mycobacteria, M. leprae and M. lepraemurium, was investigased. The results clearly demonstrated that M. paratuberculosis is a little stronger in acidfastness than tubercle bacilli, and that M. leprae and M. lepraemurium are far stronger than the latter requiring twice as much UV irradiation time as the latter for the 50% loss of acidfastness. Putting together the evidences hitherto obtained, an acidfastness map was designed. Further, very simple method to differentiate some mycobacterial strains from others by the acidfast staining was devised.
Physiological characters of twenty-four strains of Mycobacterium smegmatis, classified by a numerical classification, were investigated. The results are shown in tables 1 and 2. Colonial morphology: White or creamy, wet and smooth; non-photochromogenic. Non-motile. Acidfast rods or short rods. Growth at 3 days on egg media and on Sauton agar. Growth at 28°C, 37°C and 45°C; no growth at 52°C. Growth at 0.1% picric acid and 0.2% picric acid in Sauton agar. Growth at 0.2% Na-p-aminosalicylate and at 0.1% Na-salicylate in Ogawa egg medium. Growth at 0.25mg/ml. 8-azaguanine in Ogawa egg medium. No growth at 0.5mg/ml. NH2OH. HCl in Ogawa egg medium. Niacin: Negative. Nitrate reduction: Positive. Three-day-arylsulfatase: Negative. Two-week-arylsulfatase: Positive. PAS degradation: Negative. Salicylate degration: Negative. The following amidases are positive according to the meteod of Bönicke: Acetamidase, benzamidase, urease, isonicotinamidase, nicotinamidase, pyrazinamidase, succinamidase. The following amidase are negative in most strains: Salicylamidase, allantoinase and malonamidase. All of acetate, citrate, succinate, malate, pyruvate, benzoate, malonate and fumarate are utilized as sole carbon source for growth. The following carbohydrates are utilized as sole carbon source for growth: Glycerol, glucose, fructose, sucrose, mannose, galactose, arabinose, xylose, rhamnose, trehalose, inositol, mannitol, sorbitol, ethanol, propanol, propylene glycel, 1, 3-butylene glycol and 2, 3-butylene glycol. Raffinose and 1, 4-butylene glycol not utilized as sole carbon source for growth. Acid formed from glucose, mannose, galactose, arabinose, xylose, rhamnose, trehalose, inositol, mannitol and sorbitol, but no acid from raffinose. All of the following compounds are utilized as sole nitrogen source for growth: L-Glutamate, L-serine, L-methionine, acetamide, benzamide, urea, pyrazinamide, nicotinamide, isonicotinamide, succinamide, nitrate and nitrite. All of the following compounds are utilized as sole, simultaneous nitrogen and carbon source for growth: L-Glutamate, L-serine, glucosamine-HCl, acetamide, benzamide, monoethanolamine and trimethylene diamine. Identification. The following characters are almost specific for M. smegmatis and are considered to be “distinguishing characters” (Table 3). (1) Benzamidase; (2) isonicotinamidase; (3) succinamidase; (4) acid from galactose: (5) acid from rhamnose; (6) acid from sorbitol; (7) utilization of benzoate as sole carbon source for growth; (8) utilization of benzamide as sole nitrogen source for growth; (9) utilization of benzamide as simultaneous nitrogen and carbon source for growth.
Susceptibility of various mycobacteria to ethambutol was tested in the Löwenstein-Jensen medium. The results (Table 1) showed that the susceptibility to ethambuol be useful for differentiation of mycobacteria. Mycobacterium terrae and scotochromogens (Runyon's group II) were susceptible to 3.13μg/ml ethambutol, while M. avium and nonphotochromogens (Runyon's group III) were resistant to this concentration of ethambutol. Among rapid-growing mycobacteria and an intermediate form, M. thermoresistibile, and M. phlei and M. fortuitum were relatively resistant to ethambutol (Table 1). However, since the former two organisms can grow at 52°C and easily differentiated from other mycobacteria, test for ethambutol susceptibility of rapdily growing mycobacteria would be useful for detection of M. fortuitum, if the test is done in combination with the test for temperature range for growth.
Non-specific resistance to virulent infection with Salmonella enteritidis was studied in the mice previously infected with BCG, and the following results were obtained. 1) In CFl, DKl and dd mice, the non-specific resistance was found to be induced at the 3rd and 5th week of BCG-infection but not within one week. In DKl mice, which have been inbred in our Department as an extremely susceptible mouse strain against the infection with S. enteritidis, the non-specific resistance was induced in a low degree by BCG infection. 2) Intravenous infection with BCG was the most effective route for induction of the non-specific resistance. 3) The non-specific resistance started to be effective about 10 days after the infection with BCG and remained effective even after 13 week of the infection. 4) In BCG-infected mice growth of the infected S. enteritidis in the liver, spleen, lungs, peritoneal cavity and blood was found to be inhibited already 6 hours after the infection with Salmonella, and the growth of S. enteitidis was also found to be retarded thereafter as compared with the growth in control mice. 5) Immunization with killed BCG bacilli mixed with adjuvant could not confer the non-specific resistance on mice.