Machine learning identification ofPseudomonas aeruginosastrains from colony image data
Authors: Rattray Jennifer B. , Lowhorn Ryan J. , Walden Ryan , Márquez-Zacarías Pedro , Molotkova Evgeniya , Perron Gabriel , Solis-Lemus Claudia , Alarcon Daniel Pimentel , Brown Sam P. 
Published: September 2022

When grown on agar surfaces, microbes can produce distinct multicellular spatial structures called colonies, which contain characteristic sizes, shapes, edges, textures, and degrees of opacity and color. For over one hundred years, researchers have used these morphology cues to classify bacteria and guide more targeted treatment of pathogens. Advances in genome sequencing technology have revolutionized our ability to classify bacterial isolates and while genomic methods are in the ascendancy, morphological characterization of bacterial species has made a resurgence due to increased computing capacities and widespread application of machine learning tools. In this paper, we revisit the topic of colony morphotype on the within-species scale and apply concepts from image processing, computer vision, and deep learning to a dataset of 69 environmental and clinical Pseudomonas aeruginosa strains. We find that colony morphology and complexity under common laboratory conditions is a robust, repeatable phenotype on the level of individual strains, and therefore forms a potential basis for strain classification. We then use a deep convolutional neural network approach with a combination of data augmentation and transfer learning to overcome the typical data starvation problem in biological applications of deep learning. Using a train/validation/test split, our results achieve an average validation accuracy of 92.9% and an average test accuracy of 90.7% for the classification of individual strains. These results indicate that bacterial strains have characteristic visual ‘fingerprints’ that can serve as the basis of classification on a sub-species level. Our work illustrates the potential of image-based classification of bacterial pathogens and highlights the potential to use similar approaches to predict medically relevant strain characteristics like antibiotic resistance and virulence from colony data.

Author Summary

Since the birth of microbiology, scientists have looked at the patterns of bacterial growth on agar (colony morphology) as a key tool for identifying bacterial species. We return to this traditional approach with modern tools of computer vision and deep learning and show that we can achieve high levels of classification accuracy on a within-species scale, despite what is considered a ‘data-starved’ dataset. Our results show that strains of the environmental generalist and opportunistic pathogenPseudomonas aeruginosahave a characteristic morphological ‘fingerprint’ that enables accurate strain classification via a custom deep convolutional neural network. Our work points to extensions towards predicting phenotypes of interest (e.g. antibiotic resistance, virulence), and suggests that sample size limitations may be less restrictive than previously thought for deep learning applications in biology, given appropriate use of data augmentation and transfer-learning tools.