Different temperatures were investigated comparatively for effect on infections of the mouse foot-pad with various acid-fast bacilli.
The strains used were the Iwata, a strain of freshly isolated virulent tubercle bacilli, and the ML (M. kansasii), P5 (Group II), P25 (Group III), ATCC6841 (M. fortuitum), and ATCC927 (M. marinum), strains of atypical acid-fast bacilli. Nine or ten mice were infected with one strain after being adapted to temperatures of 10°, 20°, or 33°C in 14 days. They were inoculated subcutaneously with 10⁵-10⁶ viable organisms into the left hind foot-pad and examined daily for the appearance of foot-pad swelling. For each strain two to four mice were kept at one of the three different temperatures and sacrificed on the 14th, 28th, or 42nd day after inoculation. At autopsy gross finding on the foot-pad, lung, liver, kidney, and spleen were recorded, and each organ was cultured for viable unit count.
The optimal temperature for the occurrence of foot-pad swelling was different with the strain, as well as by the period of observation. In general, it was higher in mice kept at 10° and/or 20°C than in those kept at 33°C, except those infected with M. kansasii. In mice infected with M. kansasii, the greatest swelling was produced at an air temperature of 33°C throughout the experiment.
Acid-fast bacilli recovered from the foot-pad were larger in number in mice maintained at 10° and 20°C than in those kept at 33°C.
Gross lesions in the visceral organs were rarely encountered in mice infected with strain Iwata, P5, or ATCC6841. Viable units recovered from the visceral organs were very few, if any.
Histologically, the lesions produced in the foot-pads were granulomas.

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A collection of nonphotochromogenic, rapidly growing, acid-fast bacilli isolated from nature was analyzed on the basis of the Adansonian classification proposed by Sneath (1957).
Of ninety-one strains used, seventy-eight (86 per cent) were classified as M. fortuitum. Of these, fifty-nine of sixty-six strains studied selectively (89 per cent) were isolates: sixteen of twenty-one (76 per cent) were sewage isolates; one of one was river water isolate; two of three were sea water isolates. Most of these fresh isolates were identified as M. fortuitum. Their biological and biochemical characteristics were compatible with those of the disease-associated strains. Only two strains were recognized to beof M. smegmatis. Eleven of the remaining strains could not be classified by the methods used. No strains of M. abscessus were observed. This result suggests that this organism may hardly or not existin nature.
M. runyonii (Bojalil et al., 1962) has been listed as a synonym of M. abscessus (Moore et al., 1953). The name M. abscessus is valid, since it has priority.
There was a close relationship between M. abscessus and M. borstelense.

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Serological studies were carried out on the flagellar antigens of Vibrio parahaemolyticus by using the method of Gardner et al.
Results obtained are as follows.
1) The H antigens of 97 strains belonging to 37 serotypes of V. parahaemolyticus were identical with one another.
2) The H antigens of 18 strains belonging to V. alginolyticus were also identical with one another.
3) Reciprocal relationships were observed between the H antigens of V. parahaemolyticus and those of V. alginolyticus. Each species, however, was possessed of such H antigen factors as specific to the respective species.

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An active fraction catalyzing synthesis of dipicolinic acid from the diketopimelate-NH₃ complex was extracted from sporangium. Some physical and chemical properties of the fraction were investigated. The activity of this fraction was stable to heat treatment at 100°C for 2 hours and also to the proteinase digestion. The active fraction was dialysable against the cellophane and collodion membrane. When the effects of various reducing agents on the activity of the fraction were compared, it was found that the addition of β-mercaptoethanol to the reaction system showed a significant increase of dipicolinic acid formation. PCMB and monoiodeacetate partially inhibited the reaction. KCN completely inactivated the fraction. Neither acid hydrolysate nor ash of the fraction showed any activity.

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Calf thymus DNA, nucleic acids extracted from bacterial cells and animal tissues, amino acids, and porcine gastric mucin were tested as growth factors for mycoplasmas. The mycoplasma strains used were M. pneumoniae CL. FH, M. salivarium-C Hup 127, and M. orale N-1. These strains were kindly supplied by Dr. Hayflick.
The results obtained are as follows.
(1) DNA and extracted nucleic acids stimulated the growth of the mycoplasma strains, especially M. orale N-1. The growth of the strain was strongly influenced by the addition of nucleic acids extracted from animal tissues.
(2) Of the amino acids tested, l-glutamine, l-lysine, l-methionine, l-phenylalanine, l-threonine, and l-arginine enhanced the growth of mycoplasmas. These enhancing effects, however, should be further studied individually, because the PPLO broth tested was found to contain hitherto unknown amino acids.
(3) Porcine gastric mucin had a stimulating effect on the growth of M. salivarium and M. orale, but no effect on M. pneumoniae. The stimulating effect of the mucin was found in the fraction of acid mucopolysaccharide obtained from fractionation by means of diffusion in a Pevikon C-870 column.

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