Morphological characterisation of yeast colony growth on solid media using image processing

To rapidly determine the effect of environmental factors on yeast growth, a cell counting and colony sizing image analysis method was developed to characterise colony growth on solid media. A digitised microscopic image of the yeast was analysed using the Watershed algorithm for cell number determination and a morphological edge detection for colony size determination. The influence of temperature and physiological stress on yeast growth was then investigated over 12.5 h and data extracted by the image analysis method. © Rapid Science Ltd. 1998     

Dynamics of Leptospira colony formation on solid nutrient media]

The formation of Leptospira colonies on solid culture media varying in composition were studied. In all species of Leptospira 3 main periods of colony development were revealed, each of them having its characteristic morphological changes connected with the growth of the colony deeply into agar.     

Predictive modelling approach applied to spoilage fungi: growth of Penicillium brevicompactum on solid media

Growth of Penicillium brevicompactum was examined on five solid media. Fungal growth was established by diameter measurements up to 50 days. Seventy experimental curves were fitted by Baranyi's primary predictive model. The growth rates were then analysed by non-parametric statistical methods. Penicillium brevicompactum could colonize the surface of solid media containing up to 700 g l-1 of sugar (50% glucose-50% fructose) with a growth rate of 0.9 mm day-1 (median values). Fitting curves by non-linear models followed by a non-parametric multiple comparison seems to be a convenient method for detecting differences in fungal growth on solid media. These two methods would be useful for studying fungal spoilage of bakery products with intermediate water activity.     

Surfactant inhibition of bacterial growth on solid anthracene

Surfactants have been proposed as a promising method to enhance bioremediation of hydrophobic compounds in contaminated soils. However, the results of effects of surfactants on bioremediation are not consistent. This study showed that Triton X-100 at low concentration (0.024 mM or 0.09 CMC) inhibited the rate of growth of either a Mycobacterium sp. or a Pseudomonas sp. on solid anthracene as sole carbon source. Recovery of microbial growth rate could be achieved by dilution of surfactants, while addition of more surfactant gave an immediate decrease in growth rate. No inhibition of growth by Triton X-100 was observed with growth on glucose. The surfactant sorbed onto the surfaces of both the cells and the anthracene particles, which could inhibit uptake of anthracene. The results were consistent with the hypothesis that inhibition of microbial adhesion of cells to anthracene was responsible for the inhibition of growth by Triton X-100.     

Optimum conditions for growth of cyanobacteria on solid media

The colony forming ability of single cells or very short filaments of 7 strains of cyanobacteria was tested on media solidified by agar or by agar substitutes (Gel Gro or Gel Rite). In addition, the effect of various methods for preparation of agar media on colony forming ability was measured. High efficiency colony formation for most of the strains required that the agar be autoclaved separately from the salts in the medium. The addition of thiosulfate, but not buffer, significantly increased the plating efficiency of most strains.     

Evolution of bacterial and fungal growth media

Microbial media has undergone several changes since its inception but some key challenges remain. In recent years, there has beenexploration of several alternative nutrient sources, both to cater to the specificity in requirement of growth of “fussymicroorganisms” and also to reduce costs for large-scale fermentation that is required for biotechnology. Our mini-review exploresthese developments and also points at lacunas in the present areas of exploration, such as a lack of concerted effort in pH andosmolarity regulation. We hope that our commentary provides direction for future research in microbial media.     

Characterization of the physiological state of fungi by dynamics of colony emergence on solid media

Poisson distribution was shown to be applicable to the dynamics of emergence of fungal colonies on plates inoculated with pure cultures or environmental samples, indicating the possibility for application of Hattori approach for assessment of the physiological state of fungi. The differences in physiological activity of different fungal species and genera, between spores and mycelia, or between the fungal populations from different environments, were revealed using the t _r (delay time for colony emergence) and λ (potential capacity for growth) parameters.     

Characterization of bacterial growth on solid medium with image analysis.

Alkaliphilic bacterial strains producing the enzyme cyclodextrin glucanotransferase were cultivated on solid agar medium containing an indicator system detecting the enzyme. The growth of the colony and the surrounding diffusion zone, due to the enzyme, were measured by the image analysis during the cultivation. It was possible to differentiate between relatively similar clones by observing quantitatively the changes at and around the colony. Optimal experimental conditions for such measurements are discussed. The image analysis technique provides a potential tool for characterizing microbes grown on solid media.     

Tracking bacterial growth in liquid media and a new bacterial life model

By increasing viscosity of liquid media above 8.4 centipoise (cp) i.e. 0.084 g· cm^-1 · s^-1, individual growth and family formation ofEscherichia coli was continuously observed in real-time for up to 6 h. The observations showed primarily unidirectional growth and reproduction ofE. coli and suggested more than one reproduction in the observed portion ofE. coli life span. A new bacterial life model is proposed: each bacterium has a stable cell polarity that ultimately transforms into two bacteria of different generations; the life cycle of a bacterium can contain more than one reproduction cycle; and the age of a bacterium should be defined by its experienced chronological time. This new bacterial life model differs from the dominant concepts of bacterial life but complies with all basic life principles based on direct observation of macroorganisms.     

A computer model for the growth and differentiation of a fungal colony on solid substrate

Mold growth and differentiation are closely related to the formation of secondary products. In solid-substrate fermentations this interrelationship is often more completely realized than in submerged cultures. Solid substrate reactions are used commercially in a limited manner in the western world, but are relatively common in Asia. Basic studies in solid-substrate fermentation should yield results applicable to all types of commercial mold fermentations for the production of a secondary product. This paper presents a relatively simple model for the growth of a mold colony on a solid surface with a defined medium utilizing glucose. Unlike submerged cultures the model must account for both cellular differentiation and the spatial heterogeneity in the system. Model parameters were estimated independently using literature values. The results of the simulation studies suggest that mass transfer limitations are at least partially responsible for the proliferation of differentiated structures on solid substrates as compared to liquid cultures. Since the concentration profile depends on the depth of the substratum, conditions that enhance conidia production can be achieved by controlling the depth of the solid medium.     

Preservation of bacterial colony morphology

Summary A simple method for preserving bacterial colonies to provide a permanent plate for teaching purposes is described.     

Analysis of bacterial populations in a grassland soil according to rates of development on solid media

Abstract The process of colony formation by bacteria from grassland soil sampled in April, July and September was simulated by a colony-forming curve (CFC). The CFC was a super-imposition of several component curves (cCFC) given theoretically by the first order reaction (FOR) model [3,6]. The pattern of FOR model curves was not influenced by the time of sampling and four cCFCs were always recognized during an incubation period of 160 h. It was considered that the CFC describes an inherent property of the bacterial population of the field. Bacterial isolates were obtained from colonies produced in each of four cCFCs on agar plates. Isolates corresponding to one cCFC were classified as one group. The bacterial isolates were characterized by morphological and physiological tests and subsequently clustered. Few oligotrophic bacteria were obtained among bacteria which produced visible colonies within 63 h of incubation time. On the other hand, approx. 50% of bacteria which produced v colonies after 63 h were oligotrophic bacteria. The time required for the appearance of the first colony, t _r of the FOR model, was very similar in the isolates belonging to one group. A close linear relationship was observed between t _r value and doubling time of isolates.     

Improvements on colony morphology identification towards bacterial profiling

Colony morphology may be an indicator of phenotypic variation, this being an important adaptive process adopted by bacteria to overcome environmental stressors. Furthermore, alterations in colony traits may reflect increased virulence and antimicrobial resistance. Despite the potential relevance of using colony morphological traits, the influence of experimental conditions on colony morphogenesis has been scarcely studied in detail. This study aims to clearly and systematically demonstrate the impact of some variables, such as colony growth time, plate colony density, culture medium, planktonic or biofilm mode of growth and strain genetic background, on bacterial colony morphology features using two Pseudomonas aeruginosa strains. Results, based on 5-replicate experiments, demonstrated that all variables influenced colony morphogenesis and 18 different morphotypes were identified, showing different sizes, forms, colours, textures and margins. Colony growth time and composition of the medium were the variables that caused the highest impact on colony differentiation both derived from planktonic and biofilm cultures. Colony morphology characterization before 45 h of incubation was considered inadequate and TSA, a non-selective medium, provided more colony diversity in contrast to P. aeruginosa selective media. In conclusion, data obtained emphasized the need to perform comparisons between colony morphologies in equivalent experimental conditions to avoid misinterpretation of microbial diagnostics and biomedical studies. Since colony morphotyping showed to be a reliable method to evaluate phenotypic switching and also to infer about bacterial diversity in biofilms, these unambiguous comparisons between morphotypes may offer a quite valuable input to clinical diagnosis, aiding the decision-making towards the selection of the most suitable antibiotic and supportive treatments.     

The significances of bacterial colony patterns

Bacteria do many things as organized populations. We have recently learned much about the molecular basis of intercellular communication among prokaryotes. Colonies display bacterial capacities for multicellular coordination which can be useful in nature where bacteria predominantly grow as films, chains, mats and colonies. E. coli colonies are organized into differentiated non-clonal populations and undergo complex morphogenesis. Multicellularity regulates many aspects of bacterial physiology, including DNA rearrangement systems. In some bacterial species, colony development involves swarming (active migration of cell groups). Swarm colony development displays precise geometrical controls and periodic phenomena. Motile E. coli cells in semi-solid media form organized patterns due to chemotactic autoaggregation. On poor media, B. subtilis forms branched colonies using group motility and longrange chemical signalling. The significances of bacterial colony patterns thus reside in a deeper understanding of prokaryotic biology and evolution and in experimental systems for studying self-organization and morphogenesis.