Search

A probability matrix is presented for identification of slowly growing mycobacteria that are likely to be encountered in clinical laboratories. The matrix includes 23 features that are useful for identifying members of 14 species or species complexes. The computer program identifies strains as a function of the ID (identification) score, which measures the discrimination among possible alternative identifications, and the R (ratio) score, which measures the degree of fit to the most likely taxa. It is not necessary to employ all 23 tests when initiating an identification; the program will suggest additional tests to perform when a partial data set fails to yield a definitive identification. Two independent sets of cultures comprising a total of 1,212 strains were used to test the matrix. Correct diagnoses were based on clustering behavior in numerical taxonomic analysis with larger numbers of features. The probable efficiencies with the two sets were 94.2 and 83.4%, respectively, and the accuracy of the definitive identifications for both sets exceeded 95%. A discussion is presented of situations when it may be appropriate to override an R score that has caused the rejection of an identification and to thereby enhance the efficiency.

A total of 541 gram-negative nonfermentative bacterial strains comprising 26 species and unclassified groups were characterized by routine diagnostic and carbon substrate alkalinization techniques. These microorganisms were tested for the ability to cause alkalinization of a basal medium on a total of 217 substrates. We found that 58 carbon substrates had some discriminatory potential. We also performed 30 routine diagnostic tests. The results of these studies were evaluated by numerical taxonomy techniques. A cluster analysis of the results by the Jaccard coefficient method identified 30 clusters at the 45% level. We identified 39 tests that separated most of the groups. Groups of similar organisms or organisms that were difficult to identify were analyzed, and tests that were differential were identified. Because of variability within the clusters, further studies utilizing deoxyribonucleic acid-deoxyribonucleic acid homologies should be undertaken.

Fecal coliforms, fecal streptococci, and antimicrobial resistance (AMR) associated with various pig transit and holding times were investigated at slaughter plants. Changes in the relative abundance of two biotypes of Streptococcus faecium were associated with transit and holding of pigs, although approximately 20% of the isolates were unidentified. The greatest variety of coliforms was isolated from porcine feces after short transit (2 h) or holding (3 h) times and was qualitatively similar to those from pigs on farms. Isolates from pigs with longer average transit or holding times were almost all Escherichia coli (four biotypes). Streptococcal resistance to most antimicrobial agents was significantly greater (P less than 0.05) in isolates from live pigs at slaughter plants than in those from pigs at farms and was apparent after a short transit time (2 h). Streptococci from pigs held an average of 15 h were less resistant to most antimicrobial agents than those from pigs held 3 or 43 h. When compared with short transit times, moderate transit times (6 h) were associated with significantly decreased (P less than 0.05) coliform resistance and decreased resistance transfer but a greater diversity of AMR patterns. Holding pigs overnight (14 h) was associated with lowered coliform resistance to several antimicrobial agents, compared with the resistance of isolates from pigs held 3 or 39 h. A substantial increase (18 to 48%) in the ability to transfer streptomycin resistance was demonstrated in coliforms from pigs held 39 h, when compared with those from pigs held 3 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Previous studies have shown that Vibrio alginolyticus and Vibrio parahaemolyticus can be isolated from similar types of marine samples. In this report, the results of an examination of 567 V. alginolyticus and V. parahaemolyticus strains, isolated from seawater in Jakarta Bay and from more than 30 types of seafood from markets in Jakarta, Indonesia, are presented. Most isolates were from mackerel, shrimp, or squid. Numerical taxonomic analyses clustered 337 isolates and three V. alginolyticus reference strains at S greater than or equal to 80%. These strains produced acid from sucrose, but only approximately 80% produced acetoin or grew in the presence of 10% NaCl. The frequency of occurrence of V. alginolyticus in seawater samples ranged from 0% (in February and March 1972) to 100% (in September and December 1972) and was highest in seafood samples from August to December 1972. A second cluster of 230 isolates and seven V. parahaemolyticus reference strains was observed at S greater than or equal to 82%. These strains did not produce acetoin or acid from sucrose, and approximately 20% grew in the presence of 10% NaCl. V. parahaemolyticus was detected in seawater samples each month, with the highest frequency of occurrence (83.3%) in May 1972. Twenty-nine K antigen serotypes were demonstrated in V. parahaemolyticus isolates, and another 40% were untypable. The modal antibiotic resistance pattern for each species included five drugs. Only 12% of the V. parahaemolyticus strains were Kanagawa positive, and 10% elicited fluid accumulation in ligated rabbit ileal loops. All of the 7 V. alginolyticus strains and 94 (70%) of the V. parahaemolyticus strains tested killed mice when inoculated intraperitoneally.(ABSTRACT TRUNCATED AT 250 WORDS)

The watery exudate produced by Streptococcus mutans SL-1 colonies on sucrose-containing agar media was found to contain about 7% (wt/vol) of a water-soluble, branched dextran, 4% sucrose, and smaller (less than 1%) amounts of fructose, Folin-phenol-positive material, and lactic acid.

The distribution of polysaccharide-forming activity from sucrose was investigated in cultures of three strains of Streptococcus mutans by using an assay which conveniently determines total polysaccharide. The enzymatic activity for polysaccharide formation from sucrose is almost exclusively extracellular. The ratio of the fructan to glucan in the polysaccharide differs among the three strains investigated. The enzymatic activity for the formation of polysaccharide from sucrose has been shown to be bound to the cell-free polymer itself.

This paper describes the properties and use of a fiber optic probe as an attachment to a spectrophotometer and its use for measurements in solutions and turbid suspensions. Measurements of a standard were identical when a spectrophotometer equipped with the probe was used or when a spectrophotometer was used in a conventional manner. The probe was calibrated for turbidimetric measurements with microorganisms by relating the apparent absorbancy measured on the spectrophotometer to the dry weight of each species of organism. Continuous measurements were made of the turbidity of growing cultures of Escherichia coli, Streptococcus mutans, and Saccharomyces cerevisiae. Transient changes in cell mass were observed in some cultures during continuous monitoring of growth. The data were recovered in a manner which allowed direct computer processing.

Rogosa, M., (National Institutes of Health, Bethesda, Md.), M. I. Krichevsky, and F. S. Bishop. Truncated glycolytic system in Veillonella. J. Bacteriol. 90:164-171. 1965.-Intact Veillonella cells do not utilize carbohydrates for growth, nor are carbohydrates fermented. In cell extracts, there is no detectable glucokinase or fructokinase. Cell extracts do not degrade glucose or fructose unless supplemented with yeast hexokinase. Under these conditions, triose phosphates are formed in the presence of a hydrazine trap. When glucose-C(14) plus added hexokinase or fructose-1,6-diphosphate-C(14) was incubated with cell extracts, the production of CO(2), acetate, pyruvate, propionate, and lactate was detected. It is concluded that, except for a hexokinase, all the activities required for a glycolytic system are present.

Cultures of the amoeboid slime mold, Dictyostelium discoideum, were grown in the presence of various antibiotics and antimetabolites. Results indicate that many antibiotics may be used in amoeba cultures to help minimize contamination (for example: polymyxin B, kanamycin, tetracycline, neomycin, oxacillin, ampicillin, cephalothin, and erythromycin). A few potent antibiotics were found to be unsuitable (colistin, nystatin, amphotericin B, actinomycin, and cycloheximide); the mechanisms of the inhibitions observed were not investigated further.

A simple, sensitive, and rapid method is presented for the determination of deoxyribonucleic acid (DNA) in both gram-positive and gram-negative bacteria. It is based upon the fluorometric determination of DNA with ethidium bromide after alkaline digestion of the bacteria to hydrolyze the interfering ribonucleic acid. The assay takes less than 2 hr. Its sensitivity is at least 0.2 mug of DNA in a final solution of 4 ml and it uses commonly available filter or double monochromator fluorometers. Judicious choice of light source and filters allows an additional 10-fold increase in sensitivity with a filter fluorometer. Turbidity caused by bacteria or insoluble polysaccharides does not interfere with the fluorescence measurements. There was no significant difference between the results obtained with this method and those obtained with the indole and diphenylamine methods when these assays were applied to Escherichia coli and sucrose- or glucose-grown Streptococcus mutans. The method was also tested by determining the specific growth rate of E. coli. This new procedure should be especially useful for the determination of bacterial DNA in dilute suspensions and for the estimation of bacterial growth or DNA replication where more conventional methods are not applicable or sensitive enough.