Microbial Co-cultures Research Articles

Automated adjustment of metabolic niches enables the control of natural and engineered microbial co-cultures.

Much attention has focused on understanding microbial interactions leading to stable co-cultures. In this work, substrate pulsing was performed to promote better control of the metabolic niches (MNs) corresponding to each species, leading to the continuous co-cultivation of diverse microbial organisms. We used a cell-machine interface, which allows adjustment of the temporal profile of two MNs according to a rhythm, ensuring the successive growth of two species, in our case, a yeast and a bacterium. The resulting approach, called 'automated adjustment of metabolic niches' (AAMN), was effective for stabilizing both cooperative and competitive co-cultures. AAMN can be considered an enabling technology for the deployment of co-cultures in bioprocesses, demonstrated here based on the continuous bioproduction of p-coumaric acid. The data accumulated suggest that AAMN could be used not only for a wider range of biological systems, but also to gain fundamental insights into microbial interaction mechanisms.

Effects of combined inoculation of arbuscular mycorrhizal fungi and plant growth-promoting rhizosphere bacteria on seedling growth and rhizosphere microecology.

The effects of rhizosphere microorganisms on plant growth and the associated mechanisms are a focus of current research, but the effects of exogenous combined inoculation with arbuscular mycorrhizal fungi (AMF) and plant growth-promoting rhizobacteria (PGPR) on seedling growth and the associated rhizosphere microecological mechanisms have been little reported. In this study, a greenhouse pot experiment was used to study the effects of single or double inoculation with AM fungi (Funneliformis mosseae) and two PGPR (Bacillus sp., Pseudomonas sp.) on the growth of tobacco seedlings, together with high-throughput sequencing technology to reveal associated rhizosphere microecological mechanisms. All inoculation treatments significantly increased the aboveground dry weight; root dry weight; seedling nitrogen, phosphorus, and potassium uptake; plant height; stem thickness; maximum leaf area; chlorophyll content; total root length, surface area, and volume; and average root diameter. The highest values for these indices were observed in the combined treatment of F. mosseae and Pseudomonas sp. SG29 (A_SG29). Furthermore, the A_SG29 treatment yielded the highest diversity indexes and largest percentages of significantly enriched bacterial taxa, and significantly promoted the colonization of AMF in tobacco roots and Pseudomonas in rhizosphere soil. Differential metabolic-pathway predictions using PICRUSt2 showed that the A_SG29 treatment significantly increased the metabolic pathway richness of tobacco rhizosphere microorganisms, and significantly up-regulated some metabolic pathways that may benefit plant growth. Co-inoculation with F. mosseae and Pseudomonas sp. SG29 promoted tobacco-seedling growth by significantly improving rhizosphere microbial communities' structure and function. In summary, the combined inoculation of AMF and SG29 promotes tobacco seedling growth, optimizes the rhizosphere microbial community's structure and function, and serves as a sustainable microbial co-cultivation method for tobacco seedling production.

Regulation of the phenolic release and conversion in oats (Avena sativa L.) by co-microbiological fermentation with Monascus anka, Saccharomyces cerevisiae and Bacillus subtilis.

Microbial fermentation is an effective method to improve the functional activity of oats (Avena sativa L.), while there are some limitations to the advantages of single microbial fermentation. In this study, a microbial co-culture fermentation system with Monascus anka, Saccharomyces cerevisiae and Bacillus subtilis to release and conversion oat phenolics was established. Results showed that the optimal microbial co-fermentation system was obtained by adding Saccharomyces cerevisiae on the fourth day and Bacillus subtilis on the eighth day during Monascus anka fermentation (MF + 4S + 8B). The phenolic content was reached 26.93 mg GAE/g DW, which increased 41.08times compared to un-fermented oats (UF). In the process of co-fermentation systems, cellulase and β-glucosidase (r2 = 0.97, p < 0.01) had a positive correlation with the release of phenolics. SEM combined with HPLC showed that the complex enzyme system produced by microbial co-fermentation enhanced the disruption of oat cell structure, as well as altered the phenolics fractions and facilitated the conversion of bound phenolics to free phenolics, especially the content of chlorogenic acid and vanillic acid in the free forms was increased 31.42 and 14.15 times, respectively. Additionally, the phenolic contents were increased and the components were changed with the microbial co-fermentation of crude enzyme solution further added, which validated the positive influence of complex enzyme system of MF + 4S + 8B in the phenolic release and transformation of oats. Therefore, this study systematically investigated the phenolic mobilization in oats during the co-fermentation period, which provides a viable option for improving the functional properties of cereal products, as well as the application of microbial cell factories.

Changing the Inoculum Type From Preculture to Spore Suspension Markedly Alters the Production of Secondary Metabolites in Filamentous Microbial Coculture

The shake flask cocultures of Aspergillus terreus and Streptomyces rimosus were investigated with regard to the production of mevinolinic acid (lovastatin), oxytetracycline, and other secondary metabolites (SMs). The aim of the study was to determine the effect of inoculum type (spore suspension or preculture) on the levels of SMs in the fermentation broth. Altogether, 17 SMs were detected, including 4 products with confirmed identities, 10 putatively annotated metabolites, and 3 unknown molecules. As observed over the course of qualitative and quantitative analyses, the selection of inoculum type markedly influenced the SM-related outcomes of cocultures. Depending on the coculture initiation procedure, replacing the preculture with spore inoculum positively affected the biosynthesis of oxytetracycline, butyrolactone I, (+)-geodin, as well as the molecules putatively identified as rimocidin, CE-108, and (+)-erdin. It was concluded that the comparative analyses of SM production in filamentous microbial cocultures and monocultures are dependent on the type of inoculum and thus the diversification of inocula is highly recommended in such studies. Furthermore, it was demonstrated that designing a coculture experiment that involves only a single type of inoculum may lead to the underestimation of biosynthetic repertoires of filamentous microorganisms.

Endophytic Fungi Co-Culture: An Alternative Source of Antimicrobial Substances.

Antimicrobial resistance is becoming a critical issue due to the widespread and indiscriminate use of antibiotics and antifungals to treat common infections, leading to a growing shortage of effective drugs. Moreover, the increase in antimicrobial resistance is enhancing the pathogenicity and virulence of various pathogens. Microorganisms are key sources of chemically diverse specialized metabolites, which are produced in the final stages of their growth cycle. These metabolites hold significant value in chemical, pharmaceutical, and agrochemical industries. One of the major challenges researchers face in this field is the frequent isolation of already-known substances when classical protocols are used. To address this, several innovative strategies have been developed. The co-culture approach is a powerful tool for activating silent biosynthetic gene clusters, as it simulates natural microbial environments by creating artificial microbial communities. This method has shown promising results, with new compounds being isolated and the yields of target substances being improved. In this context, this review provides examples of antimicrobial compounds obtained from co-cultures of endophytic fungi, conducted in both liquid and solid media. Additionally, the review discusses the advantages and challenges of the co-culture technique. Significance and Impact of the Study: Microbial co-culture is a valuable strategy for discovering new natural products with antimicrobial activity, as well as for scaling up the production of target substances. This review aims to summarize important examples of endophyte co-cultures and highlights the potential of endophytic fungi co-culture for pharmacological applications.

A systematic approach to determine the outcome of the competition between two microbial species in bioreactor cocultures

The two-species microbial cocultures are effective in terms of awakening the cryptic biosynthetic pathways. They may also lead to the improved production of previously discovered molecules. Importantly, only a few outcomes of the cocultures may prove desirable, namely those leading to the formation of useful secondary metabolites. To address this issue, a method allowing for the evaluation of the final outcome of the co-culture process and fine-tune the cocultivation strategy was proposed. The systematic approach was supported by the experimental data from the bioreactor runs with the participation of Aspergillus terreus and Penicillium rubens confronted with Streptomyces rimosus and Streptomyces noursei. Kinetic, morphological and metabolic aspects of dominance were analysed via the newly proposed formula describing the dominance pattern. The suggested method involved the determination of the numerical value representing the dominance level. When it was high (value 1) no useful metabolites were formed apart from those originating from the winning counterpart. But either for the partial dominances or when the winning organism changed within the run or when the competition ended in draw, the number of the secondary metabolites of interest in the broth was the highest. Next, the systematic approach illustrated how the delayed inoculation strategy influenced the level of dominance leading to the change of winning counterpart and the set of metabolites produced. The proposed systematic approach allows for the reliable determination of the level of dominance in the two-species cocultures to seek for the potentially useful substances for future applications.

The synergistic interaction of fungi with different structural characteristics improves the leaching of lithium from lepidolite

The synergistic interaction of fungi with different structural characteristics improves the leaching of lithium from lepidolite

Insights into self-healing capacity of cement matrix containing high-efficiency bacteria under challenging conditions

Insights into self-healing capacity of cement matrix containing high-efficiency bacteria under challenging conditions

Stability of a Mutualistic Escherichia coli Co-Culture During Violacein Production Depends on the Kind of Carbon Source.

The L-tryptophan-derived purple pigment violacein (VIO) is produced in recombinant bacteria and studied for its versatile applications. Microbial synthetic co-cultures are gaining more importance as efficient factories for synthesizing high-value compounds. In this work, a mutualistic and cross-feeding Escherichia coli co-culture is metabolically engineered to produce VIO. The strains are genetically modified by auxotrophies in the tryptophan (TRP) pathway to enable a metabolic division of labor. Therein, one strain produces anthranilate (ANT) and the other transforms it into TRP and further to VIO. Population dynamics and stability depend on the choice of carbon source, impacting the presence and thus exchange of metabolites as well as overall VIO productivity. Four carbon sources (D-glucose, glycerol, D-galactose, and D-xylose) were compared. D-Xylose led to co-cultures which showed stable growth and VIO production, ANT-TRP exchange, and enhanced VIO production. Best titers were ∼126mgL-1 in shake flasks. The study demonstrates the importance and advantages of a mutualistic approach in VIO synthesis and highlights the carbon source's role in co-culture stability and productivity. Transferring this knowledge into an up-scaled bioreactor system has great potential in improving the overall VIO production.

Set Up and Utilization of a Three-Dimensional In Vitro Bioreactor System for Human Intestinal Studies and Microbial Co-Cultures.

The study of human intestinal physiology and host-microbe interactions is crucial for understanding gastrointestinal health and disease. Traditional two-dimensional cell culture models lack the complexity of the native intestinal environment, limiting their utility in studying intestinal biology. Here, we present a detailed protocol for the set up and utilization of a three-dimensional (3D) in vitro bioreactor system for human intestinal studies and bacterial co-culture. This article outlines the design and assembly of the bioreactor system, scaffold fabrication, bacterial culture techniques, analysis methods, and troubleshooting tips. By providing step-by-step instructions, the goal is to enable other laboratories to utilize physiologically relevant tissue models of the human intestine, incorporating key features, such as nutrient flow, multiple human cell types, 3D architecture, and microbial communities. The incorporation of commensal bacteria into the bioreactor system allows for the investigation of complex host-microbe interactions, providing insight into gastrointestinal health and pathology. This article serves as a comprehensive resource for scientists seeking to advance their understanding of intestinal biology toward the development of novel therapeutic strategies for gastrointestinal disorders. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Scaffold design Basic Protocol 2: Intestinal cell culture: Caco2 cells Basic Protocol 3: Intestinal cell culture: organoids Basic Protocol 4: Bioreactor design and set up Basic Protocol 5: Bacteria in 3D bioreactor set up Basic Protocol 6: Bacteria and drug dosing.

Identification and characterization of the siderochelin biosynthetic gene cluster via coculture.

Many microbial biosynthetic gene clusters (BGCs) are inactive under standard laboratory conditions, making characterization of their products difficult. Silent BGCs are likely activated by specific cues in their natural environment, such as the presence of competitors. Growth conditions such as coculture with other microbes, which more closely mimic natural environments, are practical strategies for inducing silent BGCs. Here, we utilize coculture to activate BGCs in nine actinobacteria strains. We observed increased production of the ferrous siderophores siderochelin A and B during coculture of Amycolatopsis strain WAC04611 and Tsukamurella strain WAC06889b. Furthermore, we identified the siderochelin BGC in WAC04611 and discovered that the GntR-family transcription factor sidR3 represses siderochelin production. Deletion of the predicted aminotransferase sidA abolished production of the carboxamides siderochelin A/B and led to the accumulation of the carboxylate siderochelin D. Finally, we deleted the predicted hydroxylase sidB and established that it is essential for siderochelin production. Our findings show that microbial coculture can successfully activate silent BGCs and lead to the discovery and characterization of unknown BGCs for molecules like siderochelin.IMPORTANCESiderophores are vital iron-acquisition elements required by microbes for survival in a variety of environments. Furthermore, many siderophores are essential for the virulence of various human pathogens, making them a possible target for antibacterials. The significance of our work is in the identification and characterization of the previously unknown BGC for the siderophore siderochelin. Our work adds to the growing knowledge of siderophore biosynthesis, which may aid in the future development of siderophore-targeting pharmaceuticals and inform on the ecological roles of these compounds. Furthermore, our work demonstrates that combining microbial coculture with metabolomics is a valuable strategy for identifying upregulated compounds and their BGCs.

Microbial co-cultivation: Reaction dynamics of combustible gases bioremediation in goaf

Microbial co-cultivation: Reaction dynamics of combustible gases bioremediation in goaf

Exploring microbial diversity and interactions for asbestos modifying properties

Asbestos poses a substantial environmental health risk, and biological treatment offers a promising approach to mitigate its impact by altering its chemical composition. However, the dynamics of microbial co-inoculation in asbestos bioremediation remain poorly understood. This study investigates the effect of microbial single cultures and co-cultures on modifying crocidolite and chrysotile fibers, focusing on the extraction of iron and magnesium. Seventy bacterial and eighty-three fungal strains were isolated from five diverse sites, characterized phylogenetically using the 16S rRNA gene and ITS region, respectively, and assessed for siderophore and organic acid production. Most bacterial strains were identified as Pseudomonas, while Penicillium predominated among fungal strains. Ten bacterial and 25 fungal strains were found to produce both organic compounds. Four microbial co-cultures (one bacterium-bacterium, two fungus-bacterium, and one fungus-fungus) exhibiting synergistic effects in plate assays, alongside their respective single cultures, were incubated with crocidolite and chrysotile. ICP-OES analysis revealed that in crocidolite, the co-culture HRF19-HRB12 removed more iron than their single cultures, while Penicillium TPF36 showed the highest iron removal. The co-culture of two Pseudomonas strains (HRB12-RB5) exhibited the highest magnesium concentration in the supernatant. In chrysotile, the co-culture HRB12-RB5 removed more iron than their individual cultures, with Penicillium TFSF27 exhibiting the highest iron concentration in the solution. Penicillium TFSF27 and the co-culture TFSF27-TPF36 demonstrated the highest magnesium removal. SEM-XRMA analysis showed a significant reduction in iron and magnesium content, confirming elemental extraction from the fibers' structure. This study significantly broadens the range of microbial strains capable of modifying asbestos fibers and underscores the potential of microbial co-cultures in asbestos remediation.

Bioplastic Compounds of Succinic Acid from Agriculture Waste; Date Palm Syrup And Date Palm Fronds

Excessive use of petrochemical-based plastics lead to severe plastic pollution by accumulation in soil, drainage system blockage, also each year trillions of plastic residues released into the ocean. In some cases, burning plastic waste releases toxic gases and may cause acid rain. Bioplastics, biodegradable, are eco-friendly alternatives to petroleum-based plastics, reduce environmental impacts and utilize non-edible waste products for decomposition by microorganisms, making them a significant topic in the twenty-first century. Succinic acid has wide medical and industrial application, in addition to being a solution to the problem of plastic pollution for use in the manufacture of bioplastics. One of the most important biomass wastes in the Iraqi environment that are candidates for biorefinery is palm waste due to its environmental friendliness and high sugar content. In this study, succinic acid was investigated in the fermentation product of poor-quality date palms Zahid and palm fronds. The residues of Zahid dates with palm fronds and the addition of soybean were used for the production of succinic through the process of batch fermentation and solid-state fermentation using a mixture of yeasts Saccharomyces cerevisiae and P. stipitis ATCC 58785 and fungi Trichoderma reesei and Aspergillus niger after pretreatment. The succinic acid was estimated using GC mass, and the quantities of acid produced from date juice from Zuhdi dates syrup were 20.6g/L, while the optimal ratio of acid produced from palm fronds with soybeans was estimated at 2.28g/L and production improved to 3.51 g/L by fed batch fermentation. Biotonic acid, oxalic acid malic acid were detected in significant amount as 10.82,23.68 and 70.40 g/L respectively from date syrup. This research opens the horizons for the use of the biomass of the dates palm in the production of bioplastics. Consolidated bioprocessing of lignocellulose to succinic acid through a microbial cocultivation system consolidated bioprocessing.

Biodegradation and adsorption of benzo[a]pyrene by fungi-bacterial coculture

In this work, the relationship and kinetics of biodegradation and bio-adsorption of benzo[a]pyrene (BaP) by Bacillus and Ascomycota were explored, and the metabolites of BaP under mixed microbial coculture were analyzed and characterized. The results show that BaP was removed through both biosorption and biodegradation. Under mixed microbial coculture, biosorption played a significant role in the early stage and biodegradation was predominant in the later stage. During the removal of BaP, the fungi exhibited remarkable adsorption capabilities for BaP with an adsorption efficiency (AE) of 38.14 %, while bacteria had a best degradation for BaP with a degradation efficiency (DE) of 56.13 %. Under the mixed microbial culture, the removal efficiency (RE) of BaP by the synergistic action of fungi and bacteria reached up to 76.12 % within 15 days. Kinetics analysis illustrated that the degradation and adsorption process of BaP were well fit to the first-order and the pseudo-second-order kinetic models, respectively. The research on the relationship between degradation and adsorption during microbial removal of BaP, as well as the synergistic effects of fungi and bacteria, will provide a theoretical guidance for two or even synthetic microbial communities.

Bioaugmentation protocols involving Methanobrevibacter thaueri and Pecoramyces ruminantium for investigating lignocellulose degradation and methane production from alfalfa stalks

Two protocols involving batch cultures were used to investigate the bioaugmentation of methane production by Pecoramyces ruminantium, and Methanobrevibacter thaueri. Protocol I examined the effect of altering the proportion of the microbial constituents in inoculum on alfalfa stalk fermentations and showed a 25% improvement in dry matter loss in cultures where the inoculum contained just 30% of co-culture and 70% of fungal monoculture. Protocol II involved consecutive cultures and alternating inoculations. This protocol resulted in 17-22mL/g DM methane production with co-cultures a 30% increase in methane relative to the fungal monoculture. Both protocols indicate that the co-culture rapidly dominated and was more resilient than the monoculture. Synergistic interaction between fungus and methanogen, promoted more efficient lignocellulose degradation and higher methane yield. This study highlighted the potential of microbial co-cultures for enhancing methane production from lignocellulosic biomass, offering a promising bioaugmentation strategy for improving biogas yields and waste valorization.

Synergistic enzymatic mechanism of lepidolite leaching enhanced by a mixture of Bacillus mucilaginosus and Bacillus circulans

Numerous studies have demonstrated that the co-leaching of ores by different silicate bacteria significantly improves the performance of bioleaching systems. Nevertheless, the mechanism of different silicate bacteria synergistically or complementarily enhanced the leaching process of lithium-containing silicate remains unclear. This study discussed the leaching impact of the combined presence of two metabolically distinct silicate bacteria on lepidolite, with the aim of comprehending the synergistic effect resulting from the presence of Bacillus mucilaginosus and Bacillus circulans in the leaching process. The results indicated that the polysaccharides and proteins secreted by bacteria-containing functional groups such as -OH and -COOH, which played an important role in the complex decomposition of ores. Organic acids played the role of acid etching and complexation. Bacillus mucilaginosus and Bacillus circulans exhibited low individual leaching efficiency, primarily due to their weak organic acid secretion. Moreover, the prolific polysaccharide production by Bacillus mucilaginosus led to bacterial aggregation, diminishing contact capability with minerals. Bacillus circulans decomposed the excessive polysaccharides produced by Bacillus mucilaginosus through enzymatic hydrolysis in the co-bioleaching process, providing later nutrient supply for both strains. The symbiosis of the two strains enhanced the synthesis and metabolic capabilities of both strains, resulting in increased organic acid secretion. In addition, protein and humic acid production by Bacillus mucilaginosus intensified, collectively enhancing the leaching efficiency. These findings suggested that the primary metabolic products secreted by different bacterial strains in the leaching process differ. The improvement in bioleaching efficiency during co-leaching was attributed to their effective synergistic metabolism. This work contributes to the construction of an efficient engineering microbial community to improve the efficiency of silicate mineral leaching, and reveals the feasibility of microbial co-culture to improve bioleaching.

Carbon-wise utilization of lignin-related compounds by synergistically employing anaerobic and aerobic bacteria

BackgroundLignin is a highly abundant but strongly underutilized natural resource that could serve as a sustainable feedstock for producing chemicals by microbial cell factories. Because of the heterogeneous nature of the lignin feedstocks, the biological upgrading of lignin relying on the metabolic routes of aerobic bacteria is currently considered as the most promising approach. However, the limited substrate range and the inefficient catabolism of the production hosts hinder the upgrading of lignin-related aromatics. Particularly, the aerobic O-demethylation of the methoxyl groups in aromatic substrates is energy-limited, inhibits growth, and results in carbon loss in the form of CO2.ResultsIn this study, we present a novel approach for carbon-wise utilization of lignin-related aromatics by the integration of anaerobic and aerobic metabolisms. In practice, we employed an acetogenic bacterium Acetobacterium woodii for anaerobic O-demethylation of aromatic compounds, which distinctively differs from the aerobic O-demethylation; in the process, the carbon from the methoxyl groups is fixed together with CO2 to form acetate, while the aromatic ring remains unchanged. These accessible end-metabolites were then utilized by an aerobic bacterium Acinetobacter baylyi ADP1. By utilizing this cocultivation approach, we demonstrated an upgrading of guaiacol, an abundant but inaccessible substrate to most microbes, into a plastic precursor muconate, with a nearly equimolar yields (0.9 mol/mol in a small-scale cultivation and 1.0 mol/mol in a one-pot bioreactor cultivation). The process required only a minor genetic engineering, namely a single gene knock-out. Noticeably, by employing a metabolic integration of the two bacteria, it was possible to produce biomass and muconate by utilizing only CO2 and guaiacol as carbon sources.ConclusionsBy the novel approach, we were able to overcome the issues related to aerobic O-demethylation of methoxylated aromatic substrates and demonstrated carbon-wise conversion of lignin-related aromatics to products with yields unattainable by aerobic processes. This study highlights the power of synergistic integration of distinctive metabolic features of bacteria, thus unlocking new opportunities for harnessing microbial cocultures in upgrading challenging feedstocks.

The repertoire and levels of secondary metabolites in microbial cocultures depend on the inoculation ratio: a case study involving Aspergillus terreus and Streptomyces rimosus

ObjectiveThe aim of this study was to determine the influence of the inoculation volume ratio on the production of secondary metabolites in submerged cocultures of Aspergillus terreus and Streptomyces rimosus.ResultsThe shake flask cocultures were initiated by using 23 inoculum variants that included different volumes of A. terreus and S. rimosus precultures. In addition, the axenic controls were propagated in parallel with the cocultures. UPLC‒MS analysis revealed the presence of 15 secondary metabolites, 12 of which were found both in the “A. terreus vs. S. rimosus” cocultures and axenic cultures of either A. terreus or S. rimosus. The production of the remaining 3 molecules was recorded solely in the cocultures. The repertoire and quantity of secondary metabolites were evidently dependent on the inoculation ratio. It was also noted that detecting filamentous structures resembling typical morphological forms of a given species was insufficient to predict the presence of a given metabolite.ConclusionsThe modification of the inoculation ratio is an effective strategy for awakening and enhancing the production of secondary metabolites that are not biosynthesized under axenic conditions.

Biodegradation of polyurethanes by Staphylococcus warneri and by microbial co-culture

With the increasing production and use of polyurethanes (PUs), it is necessary to develop sustainable techniques for the remediation of plastic pollution. The use of microorganisms capable of biodegrading PUs may be an environmentally desirable solution for controlling these plastic contaminants. To contribute to the discovery of alternatives for the mitigation of plastics in the environment, this study aimed to explore the potential of StaphylococcuswarneriUFV_01.21, isolated from the gut of Galleria mellonellalarvae, for biodegradation of PU in pure culture and microbial co-culture with Serratia liquefaciensL135. S. warneri grew using Impranil® PU as the sole carbon source in pure culture and co-culture. With six days of incubation, the biodegradation of Impranil® in Luria Bertani broth was 96, 88 and 76%, while in minimal medium, it was 58, 54 and 42% for S. warneri, S. liquefaciens, and co-culture, respectively. In addition, S. warneri in pure culture or co-culture was able to biodegrade, adhere and form biofilms on the surfaces of Impranil® disks and poly[4,4'-methylenebis (phenyl isocyanate)-alt-1,4-butanediol/di(propylene glycol)/polycaprolactone] (PCLMDI) films. Scanning electron microscopy also revealed biodegradation by detecting the formation of cracks, furrows, pores, and roughness on the surfaces of inoculated PU, both with pure culture and microbial co-culture. This study is the first to demonstrate the potential of S. warneriin PU biodegradation.