A Photographic Method for the Detection of Rare Bacterial Mutants in the Replica Plating Technique

Nature ◽  
1963 ◽  
Vol 199 (4889) ◽  
pp. 198-199 ◽  
Author(s):  
J. H. VAN DE POL ◽  
C. M. A. VENDRIG ◽  
G. A. VAN ARKEL
Keyword(s):  
Photographic Method ◽  
Plating Technique ◽  
Replica Plating ◽  
Bacterial Mutants

REPLICA PLATING TECHNIQUE FOR STUDYING MICROBIAL INTERACTIONS IN SOIL

1965 ◽  
Vol 11 (4) ◽  
pp. 629-636 ◽  
Author(s):  
G. Stotzky
Keyword(s):  
Nutritional Composition ◽  
Mixed Population ◽  
Microbial Interactions ◽  
Individual Species ◽  
Selective Inhibitors ◽  
Subsequent Growth ◽  
Sterile Soil ◽  
Plating Technique ◽  
Replica Plating ◽  
Plating Method

A replica plating method was developed to study ecology of microorganisms in soil. Precise placement of inocula and amendments at desired loci in sterile soil contained in petri plates were accomplished with a template. Subsequent growth and distribution of individual species, even when part of a mixed population, was measured by periodic transfer with an easily constructed replicator to agar plates of differing nutritional composition or containing selective inhibitors. The method is rapid and reproducible, and permits the study of many variables and interactions in a single soil plate; it can also be used with non-sterile soil and other suitable microbial habitats.

Changes in cutaneous flora after wet occlusion

1975 ◽  
Vol 21 (4) ◽  
pp. 496-500 ◽  
Author(s):  
David J. Bibel ◽  
Joseph R. LeBrun
Keyword(s):  
Enterobacter Aerogenes ◽  
Plating Technique ◽  
Replica Plating ◽  
Subgroup Ii ◽  
Colony Forming Units ◽  
Recovery Response ◽  
Aerobic Flora ◽  
Almost All

Aerobic flora from wet-occluded forearms of six volunteers was sampled the day before treatment, on the 3rd day when dressings were removed, and daily, when possible, for 8 days thereafter. Erythema was not present. All bacterial colonies appearing on appropriate dilution plates were identified with the aid of a replica-plating technique. Flora of each individual increased to over 104 colony-forming units/cm2 as a result of wet-occlusion, but counts rapidly fell by about 102 units once dressings were removed. Although similar types of bacteria were found on all subjects, the composition of each individual's flora during the recovery response appeared to be unique. Enterobacteriaceae were found on half the subjects with Enterobacter aerogenes being the most successful colonizer. Besides the expected presence of Baird-Parker Staphylococcus subgroup II, high numbers of subgroup IV and some colonies of subgroup III were also observed. Almost all cutaneous diphtheroids were lipophilic and lipolytic.

scholarly journals Isolation of Antibiotic Hypersusceptibility Mutants of Acinetobacter spp. by Selection for DNA Release

2003 ◽  
Vol 47 (4) ◽  
pp. 1267-1274 ◽  
Author(s):  
Hyunwoo Lee ◽  
Nora Vázquez-Laslop ◽  
Katya A. Klyachko ◽  
Alex A. Neyfakh
Keyword(s):  
Bacterial Species ◽  
Marker Gene ◽  
Opportunistic Pathogen ◽  
Bacterial Physiology ◽  
Replica Plating ◽  
Knockout Mutants ◽  
Bacterial Mutants ◽  
Dna Release ◽  
Acinetobacter Spp ◽  
Selection For

ABSTRACT Isolation of bacterial mutants hypersusceptible to antibiotics can reveal novel targets for antibiotic potentiators. However, identification of such mutants is a difficult task which normally requires laborious replica plating of thousands of colonies. The technique proposed here allows for the positive selection of genetic knockout mutants leading to hypersusceptibility. This technique, designated SDR (selection for DNA release), involves introduction of random insertions of a marker gene into the chromosome of a highly transformable bacterial species, followed by treatment of the obtained library with an antibiotic at subinhibitory concentrations. DNA released by lysing bacteria is collected and used to transform fresh bacteria, selecting for insertion of the marker gene. These selection cycles are repeated until variants with a hypersusceptibility phenotype caused by insertion of the marker begin to dominate in the library. This approach allowed for isolation of a number of mutants of the gram-negative opportunistic pathogen Acinetobacter sp. susceptible to 4- to 16-times-lower concentrations of ampicillin than wild-type bacteria. The mutations affected proteins involved in peptidoglycan turnover and, surprisingly, proteins involved in exopolysaccharide production. A further modification of the SDR technique is described which allows for selecting mutants hypersensitive to agents that affect bacterial physiology but do not cause cell lysis, e.g., inhibitors of translation. This application of SDR is illustrated here by identification of several mutants of Acinetobacter sp. with increased susceptibility (two- to fivefold decrease in the MIC) to erythromycin. The same technique can be used to identify prospective targets for potentiators of many other antibacterial agents.

scholarly journals A rapid and efficient replica plating technique

1970 ◽  
Vol 15 (3) ◽  
pp. 335-338 ◽  
Author(s):  
Thomas H. Wood ◽  
Suresh K. Mahajan
Keyword(s):  
Plating Technique ◽  
Replica Plating ◽  
Operator Error ◽  
New Type

SUMMARYA new type of apparatus for replica plating is described, which reduces operator error and fatigue. Several replica plates can be made immediately, thus eliminating the need for a master plate in many cases.

scholarly journals Alternative Replica Plating Technique

1977 ◽  
Vol 34 (2) ◽  
pp. 225-227 ◽  
Author(s):  
E. Börje Lindström
Keyword(s):  
Plating Technique ◽  
Replica Plating
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