Growth measurement of surface colonies of bacteria using augmented reality

ABSTRACT

The provision of hands-on wet lab training for students to perform micro-organism proliferation experiments is useful for experiential skills development. However, the risk of laboratory-acquired infections arising from inadvertent ingestion, inhalation or skin penetration of these micro-organisms presents a safety concern. To obviate this risk, an augmented reality exercise requiring only a tablet/smartphone, and a petri dish with appropriate markers, was devised to allow students to study Escherichia coli colony growth on agar plates. Positive responses were obtained from a pilot study conducted with high school and undergraduate students on its use.     

Growth of surface colonies of the gliding bacteriumMyxococcus xanthus

1. Colonies of several gliding, semi-motile and non-motile strains ofMyxococcus xanthus demonstrate a constant rate of diameter increase. Colonies of motile strains, characterized by a predominance of cell swarms at the leading edges, expand more rapidly than those with single, gliding cells at their peripheries.
2. Colony growth rate is proportional to the growth rate of cells in shake culture and on agar and to the square root of nutrient concentration over a wide concentration range. The rate of colony growth is affected by temperature and by agar concentration and depth but not by hight nor gravity.

     

Growth of yeast colonies on solid media

Colonies on nutrient agar of the aerobic yeast Candida utilis show linear increases in diameter and height with time throughout most of the growth cycle. The concentration of glucose in the agar has a negligible effect on radial growth rate although an increase in the glucose concentration prolongs the linear radial growth phase. The rate of increase in height of the colony is proportional to the square root of the initial glucose concentration. A new model that considers both glucose diffusion and oxygen diffusion in the colony is consistent with the observed colony profiles.     

Growth rates of yeast colonies on solid media

Previous measurements of growth rates of giant yeast colonies on solid media are shown to be unreliable as they depend strongly on extraneous factors such as the proximity of other colonies and the dimensions of the apparatus used. The hitherto unexplained dependence of the growth rate on the square root of the growth limiting nutrient concentration is explained by constructing a theory based on the diffusion of nutrient towards the colony which makes use of many ideas used in the theory of flame propagation. The theory also explains why the temperature dependence of the homogeneous growth constant is different from that observed in the surface colony, and it requires the existence of a lag phase in the homogeneous culture kinetics if the velocity of propagation of the culture is to be independent of inoculum size and shape. Both phenomena are known to occur.     

Growth patterns of yeast colonies depending on nutrient supply

Common theories of microbial growth and physiology are formulated exclusively in terms of the isolated microorganisms – especially bacteria. This is, however, an inadmissible simplification because it is obvious that the organization of microbial populations and colonies follows certain general rules. Bacterial colonies are able to generate complex interfacial growth patterns similar to those observed during diffusion-limited growth processes in non-living systems. One reason for these patterns is assumed to be the ability of many bacteria to swarm in an active manner on a substrate surface. Therefore the models of bacterial colony growth incorporate “random walkers”, which move actively in response to a gradient in the concentration of nutrients and communicate with each other by means of a chemotactic feedback. A selected number of yeasts were tested with regard to their colony growth patterns depending on the medium parameters such as nutrient concentration. Growth patterns similar to those which were described in literature for bacteria were also found in these experiments. It concerns in particular growth types like compact growth, fractal growth and dense-branching growth. This result allows a hypothesis to be formulated, that – especially in the case of fractal growth patterns – wandering of cells on a substrate surface may be induced by uncontrolled “swimming” on a thin water film caused by the metabolic activity (e.g. respiration) of the cells on the surface of the agar. Furthermore it was found that an interplay between changes in the individual morphology of yeast cells and the morphology transitions takes place. Such growth patterns are known for Candida sp. which are able to form pseudomycel and blastospores.     

Asymptotic analysis of a chemotactic model of bacteria colonies

An estimate of the distance between spots generated by a bacterial colony model is obtained. The model describes the morphogenesis of a spot pattern in colonies of chemotactic strains of Escherichia coli. Asymptotic methods for other cell-chemotaxis models, which have been successfully used by previous researchers, can be applied also to this model. However the calculations and the result is more complicated for this model. The result is verified by comparing it with the results by numerical computations of solutions of the model.     

Growth inhibition of fouling diatoms by antipatharian colonies

Growth inhibition of four dominant marine fouling diatoms by alcohol extracts of antipatharian colonies, collected off the east coast of India, are reported here. Of the three antipatharian colonies tested one antipatharian inhibited the growth of all the four fouling diatoms on glass surface, implying the presence of potent algal growth inhibitors whilst the other extracts were species specific in their action. The same cells exhibited normal growth, when transferred in extract free media, indicating a nontoxic mode of action. The effective concentration for 50% inhibition (EC50) ranged from 90 microg/ml (Nitzschia sp.) to 756 microg/ml (Amphora sp). The EC50 values for N. subinflata and N. crucicula were 215 microg/ml and 108 microg/ml, respectively. The active extracts totally arrested the silicate uptake by diatoms treated with EC100 values, in relation to controls. The degree of silicate uptake impairment varies with the test organisms. The results show the presence of antifouling agents in this group of organisms. This is the first report of anltifouling properties of antipatharian colonies.     

A method for identifying colonies of bacteria possessing mosquito larvicidal effects

Summary The method described provides a new, relatively rapid tool for the tracing or identification of B. thuringiensis var. israelensis among indigenous bacteria and even closely related variants of B. thuringiensis.     

Identification of bacteria from single colonies by fatty acid analysis.

While a commercially available system for microbial identification by fatty acid analysis (Microbial Identification System, MIDI, Newark, DE, USA) is available, it requires approximately 40-mg wet weight of biomass. Various authors have published methods for fatty acid analysis of single colonies, but no database of organisms has been developed for those methods. A modification of the MIDI system to increase sensitivity while maintaining relative retention times and peak areas allows the standard peak naming tables and libraries of organisms to be used with single colonies. Samples are evaporated down before injection to concentrate the fatty acids, while splitless injection is used to allow more sample to enter the gas chromatograph. Several known bacteria were correctly identified by this method.     

Growth of cellulolytic bacteria on sugarcane bagasse

The growth behavior of Cellulomonas has been examined in fermentation system using alkali pretreated sugarcane bagasse. During the batch operation diauxic growth was found which would not seem to be explained by catabolic repression. The relative variation of cellulose and hemicellulose during the fermentation process suggests the initial utilization of easily degradable substrate, i.e., hemicellulose and amorphous cellulose, until their concentration becomes limiting, followed by utilization of the crystalline cellulose. The conversion of substrate was 70% with a yield of 0.355 g of biomass per gram of bagasse feed.     

Growth of lithotrophic ammonia-oxidizing bacteria on hydroxylamine

Abstract A new obligately anaerobic, extremely thermophilic, cellulolytic bacterium is described. The strain designated Tp8T 6331 is differentiated from thermophilic cellulolytic clostridia on the basis of physiological characteristics and phylogenetic position within the Bacillus/Clostridium subphylum of the Gram-positive bacteria. Strain Tp8T 6331 is assigned to a new genus Caldicellulosiruptor , as Caldicellulosiruptor saccharolyticus gen., nov., sp. nov.     

Growth and characterization of T cell colonies from human thymus

A semisolid microculture system was used to study T cell colonies grown from human thymocytes. Colony growth was absolutely dependent upon media conditioned by peripheral blood leukocytes (PBL) in the presence of phytohemagglutinin. Plating efficiency was further enhanced by the addition of a non-T, adherent, radiation-resistant (7500 rad) PBL subpopulation, but was not enhanced by culture supernatants of these cells. The T colony precursor cell in the thymus occurred with a frequency of 8.0 X 10(-3) and had a surface receptor for the OKT3 monoclonal antibody. Thymocyte colony cells were functionally distinct from PBL and the major thymocyte population. The colony cells proliferated in response to T cell mitogens, but only in the presence of exogenous growth factors. The cells stimulated normal PBL in mixed leukocyte culture (MLC), but did not respond to alloantigens in MLC or in assays of spontaneous cytotoxicity. This culture system should prove helpful in the study of human thymocyte differentiation.