Abstract Manual counting of bacterial colony forming units (CFUs) on agar plates is laborious and error-prone. We therefore implemented a colony counting system with a novel segmentation algorithm to discriminate bacterial colonies from blood and other agar plates. A colony counter hardware was designed and a novel segmentation algorithm was written in MATLAB. In brief, pre-processing with Top-Hat-filtering to obtain a uniform background was followed by the segmentation step, during which the colony images were extracted from the blood agar and individual colonies were separated. A Bayes classifier was then applied to count the final number of bacterial colonies as some of the colonies could still be concatenated to form larger groups. To assess accuracy and performance of the colony counter, we tested automated colony counting of different agar plates with known CFU numbers of S. Pneumoniae, P.
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A specific amount of bacteria are reduced with every dilution. The number of colonies cultured from serial dilutions of the sample are counted to estimate the concentration of an unknown sample. Then back track the measured counts to the unknown concentration. It does not separate bacteria like a streak plate. The advantages to serial dilution-agar plate procedure is it has countable viable cells to count. As for the disadvantage of it, the techniques used which are spread and pour plates might not always have a single colony that represents the progeny of a single cell.
Aeruginosa and M. Catarrhalis and showed excellent performance.
Introduction Microbiological research techniques often rely on accurate determination of colony forming units (CFUs). Routinely, this is done by aliquoting a small amount of a liquid culture and plating out several serial dilutions onto culture plates (Petri dishes containing semisolid medium). After incubation in appropriate conditions for the microorganism of choice, the colonies are counted to determine the number of CFU. This is done by manually counting of colonies on plates illuminated by transmitted light. The concentration of bacteria in the original culture can then be calculated based on the assumption that each colony has raised from one single bacterium (colony forming unit, CFU). This process is time-consuming, tedious and error prone.
There is a tendency to analyse only high dilutions of the initial culture as these have fewer colonies to count. Unfortunately, in low count assays minor counting errors have significant effects on the calculated concentration in the primary liquid medium. On the other hand, accurate counting of plates with high numbers of CFUs is error prone since it requires a high level of attention by the performer. Therefore, often only parts of a plate are analyzed and used to estimate the whole plate count after extrapolation. Furthermore high numbers of CFUs on a plate can lead to false redults due to overcrowding of bacteria. This study aimed to design an automated colony counter which reliably detects, bacterial counts and colony size on semisolid agar plates of 85 mm diameter.
The system should be suitable for the study of important human pathogens, such as Streptococcus pneumoniae, Moraxella catarrhalis and Pseudomonas aeruginosa. These bacteria are grown on diverse agar plates including Columbia blood sheep agar (CSBA), chocolate agar and brain heart infusion (BHI) agar plates. The system should also be user-friendly and cost-effective with an algorithm that is adaptable to other culture media and microorganisms. Bacterial strains and growth conditions A total of 7 clinical isolates of S.