Beyond Cultivation: Combining Culture-Dependent and Culture-Independent Techniques to Identify Bacteria Involved in Paint Spoilage

Abstract

Due to globally increasing problems concerning biodeterioration of paints, it is worthwhile to enhance the determination of colony forming units (CFU) as a gold standard method via more rapid and culture-independent techniques. Here, we combined traditional culture-dependent techniques with subsequent sequencing, quantitative qPCR, and a serial quantification method (most probable number; MPN) to detect paint degrading bacteria in general and sulfate-reducing bacteria (SRB) in particular. During our investigation of three water-based paints that showed visible contamination, we found high bacterial counts of up to 107 CFU mL−1. Subsequent sequencing allowed the identification of common paint degraders including Bacillus sp. and Pseudomonas sp., but less frequently detected bacteria such as Rhodococcus sp. and Delftia spp. were also found to be present. MPN, as well as dsrA-targeted qPCR to detect SRB, only showed positive results for two out of three samples. These results coincided with the inherent physicochemical properties of the paints offering suitable conditions for microbial growth or not. The MPN method can be used for a diversity of aerobic and anaerobic bacteria and is rapid and reproducible. A combination of culture-independent techniques such as qPCR or NGS can help to fully elucidate the bacterial diversity in spoiled paint by also recovering anaerobic and unculturable ones.

Toward Self-Healing Coatings: Bacterial Survival and Calcium Carbonate Precipitation in Acrylic and Styrene–Acrylic Model Paint Films

“…While much work has attempted to identify the species responsible for this growth, a range of species have been observed, and no work has attempted to identify the factors that allow for survival and growth in these chemistries [43]. Instead, most research (and patents) examining microbial survival in paint formulations has been geared toward limiting microbial (bacterial and fungal) survival and growth, either through the addition of biocides or formulation [41,44,45]. Our approach was to determine whether we could leverage microbial growth to generate a unique, engineered living material approach to improve the functionality of paint, while attempting to understand some of the factors that allow survival in a type bacterial species.…”

Breaking the Mold: Towards Rapid and Cost-Effective Microbial Contamination Detection in Paints and Cosmetics Using ATP-Bioluminescence

“…The latter comprises polymerase chain reaction (PCR), quantitative PCR (qPCR), digital PCR (dPCR), high-throughput next-generation DNA sequencing, matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) MS, ion mobility spectrometry (IMS) or determination of biomarker volatile organic compounds (MVOC) by solid-phase microextraction gas chromatography-mass spectrometry [5][6][7][8][9]. Matching with cultureindependent methods, we succeeded in employing qPCR and most-probable number techniques for detecting paint-spoiling bacteria in general and sulfate-reducing bacteria in particular in a previous investigation [10]. Alternative methodologies to classical microbiological counting are intended to save materials and time.…”