Water Biofilms Research Articles

Anti-biofilm, drug delivery and cytotoxicity properties of dendrimers.

Treatments using antimicrobial agents have faced many difficulties as a result of biofilm formation by pathogenic microorganisms. The biofilm matrix formed by these microorganisms prevents antimicrobial agents from penetrating the interior where they can exact their activity effectively. Additionally, extracellular polymeric molecules associated with biofilm surfaces can absorb antimicrobial compounds, lowering their bioavailability. This problem has resulted in the quest for alternative treatment protocols, and the development of nanomaterials and devices through nanotechnology has recently been on the rise. The literature on dendrimers was searched for in databases such as Google Scholar, PubMed, and ScienceDirect. As a nanomaterial, dendrimers have found useful applications as a drug delivery vehicle for antimicrobial agents against biofilm-mediated infections to circumvent these defense mechanisms. The distinctive properties of dendrimers, such as multi-valency, biocompatibility, high water solubility, non-immunogenicity, and biofilm matrix-/cell membrane fusogenicity (ability to merge with intracellular membrane or other proteins), significantly increase the efficacy of antimicrobial agents and reduce the likelihood of recurring infections. This review outlines the current state of dendrimer carriers for biofilm treatments, provides examples of their real-world uses, and examines potential drawbacks.

Responses of drinking water bulk and biofilm microbiota to elevated water age in bench-scale simulated distribution systems

Reductions in nonresidential water demand during the COVID-19 pandemic highlighted the importance of understanding how water age impacts drinking water quality and microbiota in piped distribution systems. Using benchtop model distribution systems, we aimed to characterize the impacts of elevated water age on microbiota in bulk water and pipe wall biofilms. Five replicate constant-flow reactors were fed with municipal chloraminated tap water for 6 months prior to building closures and 7 months after. After building closures, chloramine levels entering the reactors dropped; in the reactor bulk water and biofilms the mean cell counts and ATP concentrations increased over an order of magnitude while the detection of opportunistic pathogens remained low. Water age, and the corresponding physicochemical changes, strongly influenced microbial abundance and community composition. Differential initial microbial colonization also had a lasting influence on microbial communities in each reactor (i.e., historical contingency).

Emerging investigator series: optimisation of drinking water biofilm cell detachment and sample homogenisation methods for rapid quantification via flow cytometry

Establishing an optimized method to harvest and quantify cells of biofilms from drinking water systems.

Revealing the characteristics of biofilms on different polypropylene plastic products: Comparison between disposable masks and takeaway boxes

The increasingly severe plastic pollution issue was intensified by the enormous plastic emissions into ecosystems during the Covid-19 pandemic. Plastic wastes entering the environment were swiftly exposed to microorganisms and colonized by biofilms, and the plastic-biofilm combined effects further influenced the ecosystem. However, the non-woven structure of disposable masks discarded carelessly during the COVID-19 pandemic was different from those of plastics with flat surface. To reveal the potential effects of plastic structure on colonized biofilms, white disposable surgical masks (DM) and transparent takeaway boxes (TB), both made of polyethylene, were selected for the incubation of organic conditioning films and biofilms. The results indicated that the non-woven structure of disposable mask was destroyed by the influence of water infiltration and biofilm colonization. The influence of surface structure on conditioning films led to a relatively higher proportion of tryptophan-like substances on DM than those on TB samples. Therefore, biofilms with significantly higher microbial biomass and carbon metabolic capacity were formed on DM than those on TB samples owing to the combined effects of their differences in surface structure and conditioning films. Moreover, abundant functional microorganisms associated with stress tolerance, carbon metabolism and biofilm formation were observed in biofilms on disposable mask. Combining with the results of partial least squares regression analysis, the selective colonization of functional microorganisms on disposable masks with uneven surface longitudinal fluctuation was revealed. Although the predicted functions of biofilms on disposable masks and takeaway boxes showed more similarity to each other than to those of free-living aquatic microorganisms owing to the existence of the plastisphere, biofilms on disposable masks may potentially trigger environmental risks different from those of takeaway boxes by unique carbon metabolism and abundant biomass.

Prospective healthcare-associated links in transmission of nontuberculous mycobacteria among people with cystic fibrosis (pHALT NTM) study: Rationale and study design.

Healthcare-associated acquisition and transmission of nontuberculous mycobacteria (NTM) among people with cystic fibrosis (pwCF) has been described, and remains a concern for both patients and providers. This report describes the design of a prospective observational study utilizing the standardized epidemiologic investigation toolkit for healthcare-associated links in transmission of NTM among pwCF. This is a parallel multi-site study of pwCF who have infections with respiratory NTM isolates and receive healthcare within a common CF Care Center. Participants have a history of one or more NTM positive airway cultures and have been identified as having NTM infections suggestive of a possible outbreak within a single Center, based on NTM isolate genomic analysis. Participants are enrolled in the study over a 3-year period. Primary endpoints are identification of shared healthcare-associated source(s) among pwCF in a Center, identification of healthcare environmental dust and water biofilm NTM isolates that are genetically highly-related to respiratory isolates, and identification of common home of residence watersheds among pwCF infected with clustered isolates. Secondary endpoints include characterization of healthcare-associated transmission and/or acquisition modes and settings as well as description of incidence and prevalence of healthcare-associated environmental NTM species/subspecies by geographical region. We hypothesize that genetically highly-related isolates of NTM among pwCF cared for at the same Center may arise from healthcare sources including patient-to-patient transmission and/or acquisition from health-care environmental dust and/or water biofilms. This study design utilizes a published, standardized, evidence-based epidemiologic toolkit to facilitate confidential, independent healthcare-associated NTM outbreak investigations within CF Care Centers. This study will facilitate real-time, rapid detection and mitigation of healthcare-associated NTM outbreaks to reduce NTM risk, inform infection prevention and control guidelines, and characterize the prevalence and origin of NTM outbreaks from healthcare-associated patient-to-patient transmission and/or environmental acquisition. This study will systematically characterize human disease causing NTM isolates from serial collection of healthcare environmental dust and water biofilms and define the most common healthcare environmental sources harboring NTM biofilms. ClinicalTrials.gov NCT05686837.

Reshaping the plastisphere upon deposition: Promote N2O production through affecting sediment microbial communities in aquaculture pond

Microplastics (MPs) could provide vector for microorganisms to form biofilm (plastisphere), but the shaping process of MPs biofilm and its effects on the structure and function of sedimentary microbial communities especially in aquaculture environments are not reported. For this, we incubated MPs biofilm in situ in an aquaculture pond and established a sediment microcosm with plastisphere. We found that the formation of MPs biofilm in surface water was basically stable after 30 d incubation, but the biofilm communities were reshaped after deposition for another 30 d, because they were more similar to plastisphere communities incubated directly within sediment but not surface water. Moreover, microbial communities of MPs-contaminated sediment were altered, which was mainly driven by the biofilm communities present on MPs, because they but not sediment communities in proximity to MPs had a more pronounced separation from the control sediment communities. In the presence of MPs, increased sediment nitrification, denitrification and N2O production rates were observed. The K00371 (NO2-⇋NO3-) pathway and elevated abundance of nxrB and narH genes were screened by metagenomic analysis. Based on structural equation model, two key bacteria (Alphaproteobacteria bacterium and Rhodobacteraceae bacterium) associated with N2O production were further identified. Overall, the settling of MPs could reshape the original biofilm and promote N2O production by selectively elevating sedimental microorganisms and functional genes in aquaculture pond.

Freshwater algal biofilm assemblages are more effective than invertebrate assemblages at aggregating microplastics

Microplastics, plastic particles less than 5 mm in length, are a ubiquitous pollutant in the environment, but research on freshwater microplastic contamination is lacking. A possible fate of microplastics in freshwater environments is to become entangled or aggregated in biofilms, which are matrices of algae, bacteria, and micro invertebrates that grow on underwater surfaces, following a progression of settling algae, periphyton, and finally invertebrate colonization. This in-situ study at the Oasis Marina at National Harbor in Oxon Hill, Maryland, examined how the taxonomic assemblages of freshwater biofilms in the Potomac River are associated with the number of microplastics aggregated within them. Aluminum discs, acting as artificial substrate for biofilm growth, were deployed at the water's surface and at 2 m depth to survey biofilm assemblage and were sampled monthly from October 2021–October 2022. Microplastic abundances in the water column were measured every 2 weeks over the same period. Spatial and temporal trends in trapped and suspended microplastics, water quality parameters (temperature, dissolved oxygen, pH, salinity, conductivity, turbidity, ammonia, nitrate, and phosphate), and biofilm assemblages were measured and compared to explore factors affecting the abundance of microplastics and their partitioning between the water column and biofilms. Water quality had no measurable impact on microplastic abundance in the water column at either depth, but temperature was negatively correlated to microplastic abundance in biofilms. As the weather warmed and biofilms progressed to invertebrate settling, they tended to contain fewer microplastics. This may have occurred because less biologically rich biofilms, primarily composed of unicellular algal colonies, provide a favorable surface for microplastic deposition. Understanding seasonal changes in biofilm assemblage and microplastic abundance may help track the fate of microplastics in freshwater systems, particularly in their interactions with lower trophic organisms.

Draft genome sequence of Sphingomonas paucimobilis strain Sph5, isolated from tap water filtration membrane.

Sphingomonadaceae are common membrane colonizers and biofilm formers. As part of our studies on long-term genetic changes in drinking water biofilm species, we report the draft genome sequence of Sphingomonas strain Sph5, isolated from a tap water filtration membrane. The isolate was determined as Sphingomonas paucimobilis through whole genome sequencing and de novo assembly.

Algae biofilm induced surface erosion of acrylic coatings

A major factor influencing failures of water-based elastomeric roof coatings is extended exposure to ponding water, biofilm development, attachment, and damage. Development of improved coatings is slow due to outdoor exposure tests that can take years. The goal of this work is to develop accelerated test methods and to understand the mechanism of biofilm-induced damage of coatings. Two model acrylic coatings on TPO substrates were exposed to lab-controlled accelerated growth of biofilm (algae) in ponding water to learn about coating damage before and after subsequent wet-dry cycles. Three distinct damage features in sequence were observed: (1) pit formation on the coating surfaces, (2) the biofilm mud-cracking induced coating surface damage upon drying, and (3) erosion (surface cracks) of coating in the mud-cracked region over the wet-dry cycles. Drying of adequately bonded biofilm to the coating and the thickness of the biofilm are the critical factors that govern the erosion on the coating surfaces. The effect of the wet-dry cycle time period was studied with respect to the surface erosion of the coatings. The mechanisms of biofilm-induced erosion during wet-dry cycling are postulated based on the mechanics and experimental observations. A new accelerated testing methodology for evaluating long-term roof coating performance is proposed.

The Role of Chlorine in the Formation and Development of Tap Water Biofilms under Different Flow Regimes.

Water companies make efforts to reduce the risk of microbial contamination in drinking water. A widely used strategy is to introduce chlorine into the drinking water distribution system (DWDS). A subtle potential risk is that non-lethal chlorine residuals may select for chlorine resistant species in the biofilms that reside in DWDS. Here, we quantify the thickness, density, and coverage of naturally occurring multi-species biofilms grown on slides in tap water with and without chlorine, using fluorescence microscopy. We then place the slides in an annular rotating reactor and expose them to fluid-wall shears, which are redolent of those on pipe walls in DWDS. We found that biofilms in chlorine experiment were thicker, denser and with higher coverage than in non-chlorine conditions under all flow regimes and during incubation. This suggests that the formation and development of biofilms was promoted by chlorine. Surprisingly, for both chlorinated and non-chlorinated conditions, biofilm thickness, density and coverage were all positively correlated with shear stress. More differences were detected in biofilms under the different flow regimes in non-chlorine than in chlorine experiments. This suggests a more robust biofilm under chlorine conditions. While this might imply less mobilization of biofilms in high shear events in pipe networks, it might also provide refuge from chlorine residuals for pathogens.

Modelling drinking water biofilms: Bacterial adhesion and Legionella pneumophila necrotrophic growth

We propose a mathematical model to simulate the establishment and growth of a drinking water distribution system biofilm, focusing on the influence of ionic strength on bacterial adhesion and persistence of Legionella pneumophila. Specifically, we consider how ionic strength affects interaction energies during the initial phase of biofilm formation and we include the dependence of the attachment flux, modelled as a linear rate with respect to free floating cells, on it. The model also incorporates a novel necrotrophic kinetics to simulate the Legionella pneumophila metabolism. The biofilm is modelled as a 1D free boundary domain, and its evolution is governed by hyperbolic–parabolic PDEs. The initial attachment phase is modelled by considering a vanishing initial value for the free boundary. The model is investigated numerically highlighting the impact of the necrotrophic kinetic parameters, the influence of ionic strength on the initial stage of biofilm formation through bacterial attachment, and how variations in nutrient levels affect system dynamics. The numerical results demonstrate that: the ionic strength mainly governs bacterial adhesion for young biofilms and affects the biofilm ecology also in presence of a biocide; the necrotrophic metabolism of Legionella pneumophila favours its persistence in oligotrophic biofilms.

Ultraefficient OG-Mediated Photodynamic Inactivation Mechanism for Ablation of Bacteria and Biofilms in Water Augmented by Potassium Iodide under Blue Light Irradiation.

While photodynamic inactivation (PDI) has emerged as a novel sterilization strategy for drinking water treatment that recently attracted tremendous attention, its efficiency needs to be further improved. In this study, we aimed to clarify the ultraefficient mechanism by which potassium iodide (KI) potentiates octyl gallate (OG)-mediated PDI against bacteria and biofilms in water. When OG (0.15 mM) and bacteria were exposed to blue light (BL, 420 nm, 210 mW/cm2), complete sterilization (>7.5 Log cfu/mL of killing) was achieved by the addition of KI (250 mM) within only 5 min (63.9 J/cm2). In addition, at lower doses of OG (0.1 mM) with KI (100 mM), the biofilm was completely eradicated within 10 min (127.8 J/cm2). The KI-potentiated mechanism involves in situ rapid photogeneration of a multitude of reactive oxygen species, especially hydroxyl radicals (•OH), reactive iodine species, and new photocytocidal substances (quinone) by multiple photochemical pathways, which led to the destruction of cell membranes and membrane proteins, the cleavage of genomic DNA and extracellular DNA within biofilms, and the degradation of QS signaling molecules. This multitarget synergistic strategy provided new insights into the development of an environmentally friendly, safe, and ultraefficient photodynamic drinking water sterilization technology.

Aquaculture ecosystem microbiome at the water-fish interface: the case-study of rainbow trout fed with Tenebrio molitor novel diets

BackgroundSustainable aquaculture relies on multiple factors, including water quality, fish diets, and farmed fish. Replacing fishmeal (FM) with alternative protein sources is key for improving sustainability in aquaculture and promoting fish health. Indeed, great research efforts have been made to evaluate novel feed formulations, focusing especially on the effects on the fish gut microbiome. Few studies have explored host-environment interactions. In the present study, we evaluated the influence of novel insect-based (Tenebrio molitor) fish diets on the microbiome at the water-fish interface in an engineered rainbow trout (Oncorhynchus mykiss) farming ecosystem. Using 16S rRNA gene metabarcoding, we comprehensively analyzed the microbiomes of water, tank biofilm, fish intestinal mucus, fish cutis, and feed samples.ResultsCore microbiome analysis revealed the presence of a highly reduced core shared by all sample sources, constituted by Aeromonas spp., in both the control and novel feed test groups. Network analysis showed that samples were clustered based on the sample source, with no significant differences related to the feed formulation tested. Thus, the different diets did not seem to affect the environment (water and tank biofilm) and fish (cutis and intestinal mucus) microbiomes. To disentangle the contribution of feed at a finer scale, we performed a differential abundance analysis and observed differential enrichment/impoverishment in specific taxa, comparing the samples belonging to the control diet group and the insect-based diet group.ConclusionsOmic exploration of the water-fish interface exposes patterns that are otherwise undetected. These data demonstrate a link between the environment and fish and show that subtle but significant differences are caused by feed composition. Thus, the research presented here is a step towards positively influencing the aquaculture environment and its microbiome.

Influence of High-Frequency, Low-Voltage Alternating Electric Fields on Biofilm Development Processes of Escherichia coli and Pseudomonas aeruginosa

A non-chemical solution is needed to control biofilms in water and wastewater treatment systems. High-frequency alternating electric field application offers an alternative approach that does not involve undesired electrode surface reactions. However, the effect of high-frequency alternating electric fields on bacterial cells in the attached-growth mode remains unexplored. This study investigated the impact of such fields on two stages of the biofilm development process: the initial adhesion phase (stage 1) and the early development phase (stage 2). Experiments were conducted using Escherichia coli and Pseudomonas aeruginosa in a three-channel flow cell exposed to alternating electric fields (3.05 V/cm, 20 MHz). The primary outcome of this study demonstrated that alternating electric fields decreased adhered cell numbers at both stages due to their inhibitory effect on growth. The alternating electric fields also triggered cell detachment after the initial attachment stage but not in mature biofilms. Interestingly, despite a reduction in cell counts, the amount of total biofilm biomass remained unaffected, which was likely due to increased cell size via cell elongation compensating for the decrease in numbers. No synergistic effects with respect to hydrodynamic forces were observed. These findings highlight the potential applicability of alternating electric fields to biofilm control and provide implications for water and wastewater engineering applications.

Evaluation of protein extraction methods to improve meta-proteomics analysis of treated wastewater biofilms.

Metaproteomics can be used to study functionally active biofilm-based bacterial populations in reclaimed water distribution systems, which in turn result in bacterial regrowth that impacts the water quality. However, existing protein extraction methods have differences in their protein recovery and have not been evaluated for their efficacies in reclaimed water biofilm samples. In this study, we first evaluated six different protein extraction methods with diverse chemical and physical properties on a mixture of bacterial cell culture. Based on a weighting scores-based evaluation, the extraction protocols in order of decreasing performance are listed as B-PER>RIPA>PreOmics>SDS>AllPrep>Urea. The highest four optimal methods on cell culture were further tested against treated wastewater non-chlorinated and chlorinated effluent biofilms. In terms of protein yield, our findings showed that RIPA performed the best; however, the highest number of proteins were extracted from SDS and PreOmics. Furthermore, SDS and PreOmics worked best to rupture gram-positive and gram-negative bacterial cell walls. Considering the five evaluation factors, PreOmics obtained highest weighted score, indicating its potential effectiveness in extracting proteins from biofilms. This study provides the first insight into evaluating protein extraction methods to facilitate metaproteomics for complex reclaimed water matrices.

Effect of aquaponic water and substratum material on biofilm formation by Aeromonas hydrophila

Aeromonas hydrophila is a zoonotic pathogen causing illness in fish and susceptible humans. This emerging pathogen has been isolated within aquaponic systems and could cause disease in fish and a hazard to humans consuming aquaponic produce. This study determined whether A. hydrophila from an aquaponic farm could form biofilms in aquaponic water and on materials used in these systems. A. hydrophila biofilm biomass and cell density in aquaponic water were evaluated by crystal violet staining and culture-based enumeration. Biofilm biomass and biofilm cell density were affected by the water source and A. hydrophila isolate (P < 0.05). A. hydrophila formed the most biomass from the beginning of deep-water culture (BDWC) water (OD570 0.202 ± 0.066) and the least from the end of deep-water culture (EDWC) water (OD570 0.140 ± 0.036; P < 0.05). Enumerated A. hydrophila from the biofilm varied among water sources; the fish tank water supported the greatest cell density (7.04 ± 0.71 log CFU/mL) while the EDWC supported the lowest cell density (6.76 ± 0.83 log CFU/mL). Biofilm formation was also evaluated on aquaponic materials such as nylon, polyvinyl chloride, polyethylene liner, bead filter, and foam. Biofilm formation on the liner had the greatest population (2.39 ± 0.022 log CFU/cm2), and the bead had the least (0.64 ± 0.039 log CFU/cm2; P < 0.05). Pathogenic organisms, such as A. hydrophila, may pose a greater risk to produce harvested from the BDWC and MDWC due to greater biofilm formation.

Influence of Temperature on Growth of Four Different Opportunistic Pathogens in Drinking Water Biofilms.

High drinking water temperatures occur due to climate change and could enhance the growth of opportunistic pathogens in drinking water systems. We investigated the influence of drinking water temperatures on the growth of Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Mycobacterium kansasii and Aspergillus fumigatus in drinking water biofilms with an autochthonous microflora. Our results reveal that the growth of P. aeruginosa and S. maltophilia in the biofilm already occurred at 15.0 °C, whereas M. kansasii and A. fumigatus were able to grow when temperatures were above 20.0 °C and 25.0 °C, respectively. Moreover, the maximum growth yield of P. aeruginosa, M. kansasii and A. fumigatus increased with increasing temperatures up to 30 °C, whereas an effect of temperature on the yield of S. maltophilia could not be established. In contrast, the maximum ATP concentration of the biofilm decreased with increasing temperatures. We conclude from these results that high drinking water temperatures caused by, e.g., climate change can result in high numbers of P. aeruginosa, M. kansasii and A. fumigatus in drinking water systems, which poses a possible risk to public health. Consequently, it is recommended for countries with a more moderate climate to use or maintain a drinking water maximum standard temperature of 25 °C.

The consistent application of hydrogen peroxide controls biofilm growth and removes Vermamoeba vermiformis from multi-kingdom in-vitro dental unit water biofilms

The water systems inside a dental unit are known to be contaminated with a multi-kingdom biofilm encompassing bacteria, fungi, viruses and protozoa. Aerosolization of these micro-organisms can potentially create a health hazard for both dental staff and the patient. Very little is known on the efficacy of dental unit disinfection products against amoeba. In this study we have examined the effect of four different treatment regimens, with the hydrogen peroxide (H2O2) containing product Oxygenal, on an in-vitro multi-kingdom dental unit water system (DUWS) biofilm. The treatment efficacy was assessed in time using heterotrophic plate counts, the bacterial 16S rDNA, fungal 18S rDNA gene load and the number of genomic units for Legionella spp. the amoeba Vermamoeba vermiformis. The results indicated that a daily treatment of the DUWS with a low dose H2O2 (0.02% for 5 h), combined with a weekly shock dose (0.25% H2O2, 30 min) is necessary to reduce the heterotrophic plate count of a severely contaminated DUWS (>106 CFU.mL−1) to below 100 CFU.mL−1. A daily treatment with a low dose hydrogen peroxide alone, is sufficient for the statistically significant reduction of the total amount of bacterial 16S rDNA gene, Legionella spp. and Vermamoeba vermiformis load (p < 0.005). Also shown is that even though hydrogen peroxide does not kill the trophozoite nor the cysts of V. vermiformis, it does however result in the detachment of the trophozoite form of this amoeba from the DUWS biofilm and hereby ultimately removing the amoeba from the system.

Induction of bacteria in biofilm into a VBNC state by chlorine and monitoring of biofilm structure changes by means of OCT

The occurrence of viable but non-culturable (VBNC) bacteria in drinking water may result in significant underestimation of viable cell counts detected by culture-based method, thus raising microbiological safety concern. Chlorine disinfection has been widely used in drinking water treatment to ensure microbiological safety. However, the effect of residual chlorine on inducing bacteria in biofilms into a VBNC state remains unclear. We determined cell numbers of Pseudomonas fluorescence in different physiological states (culturable, viable, dead) by heterotrophic plate count method and flow cytometer in a flow cell system under 0, 0.1, 0.5, 1.0 mg/L chlorine treatment. Numbers of culturable cells were 4.66 ± 0.47 Log10, 2.82 ± 0.76 Log10, 2.30 ± 1.23 Log10 (CFU/112.5 mm3) in each chlorine treatment group. However, viable cell numbers remained at 6.32 ± 0.05 Log10, 6.11 ± 0.24 Log10, 5.08 ± 0.81 Log10 (cells/112.5 mm3). Significant difference between numbers of viable and culturable cells demonstrated chlorine could induce bacteria in biofilms into a VBNC state. In this study, flow cells combination with Optical Coherence Tomography (OCT) were applied to construct an Automated experimental Platform for replicate Biofilm cultivation and structural Monitoring (APBM) system. The OCT imaging results demonstrated that changes of biofilm structure under chlorine treatment were closely related to their inherent characteristics. Biofilms with low thickness and high roughness coefficient or porosity were easier to be removed from the substratum. Biofilm with high rigid properties were more resistant to chlorine treatment. Even though >95 % bacteria in biofilms entered a VBNC state, the biofilm physical structure was still remained. This study revealed the possibility of bacteria to enter a VBNC state in drinking water biofilms and changes of biofilm structure with different characteristics under chlorine treatment, which provide reference for biofilms control in drinking water distribution systems.

Bacterial diversity and co‐occurrence patterns differ across a world‐wide spatial distribution of habitats in glacier ecosystems

Abstract Glaciers cover nearly 10% of the Earth's surface and are unique biomes dominated by microbial communities that support key ecosystem processes. The melting of glaciers is among the most conspicuous consequences of global climate change, with impacts on microbial ecology and associated biogeochemistry. However, we are still missing an integrative understanding of microbial biodiversity from divergent habitats associated with glaciers. Here, we compiled global microbiome metadata from 93 representative glaciers over 180 locations and used random forest, microbial ecological networks and structural equation modelling to evaluate the biodiversity and environmental factors associated with the glacier microbiomes of seven contrasting habitats: water, epilithic biofilm, cryoconite, mat, ice, sediment and permafrost soil. The results showed that microbial diversity largely changed across habitats, with the highest in permafrost soil, followed by sediment, ice, mat, cryoconite, water and epilithic biofilm. More importantly, we provided critical evidence that the environmental and climatic factors associated with the microbiomes of glaciers varied with glacier habitats. Microbial diversity in water was highly correlated with latitude, cryoconite microbial diversity was significantly (p = 0.01) correlated with pH, and permafrost soil and sediment microbial diversity were mainly explained by temperature (17.05% and 13.37% respectively). Using ecological association network analysis, we identified some tightly linked common microbial taxa (e.g. Proteobacteria and Bacteroidetes) that were present in all the habitats and were vulnerable to climatic factors, such as temperature and precipitation. This study demonstrated that microbial diversity, drivers and co‐occurrence patterns differ among glacier habitats globally, and diverse habitat‐dependent glacier microbiomes could serve as early warning sentinels for the study of life on glaciers and its potential future in a warming world. Read the free Plain Language Summary for this article on the Journal blog.