Extracellular DNA (eDNA) and intracellular DNA (iDNA) extensively exist in both terrestrial and aquatic environment systems and have been found to play a significant role in the nutrient cycling and genetic information transmission between the environment and microorganisms. As inert DNA sequences, eDNA is able to present stability in the environment from the ribosome enzyme lysis, therein acting as the historical genetic information archive of the microbiome. As a consequence, both eDNA and iDNA can shed light on the functional gene variety and the corresponding microbial activity. In addition, eDNA is a ubiquitous composition of the cell membrane, which exerts a great impact on the resistance of outer stress from environmental pollutants, such as heavy metals, antibiotics, pesticides, and so on. This study focuses on the environmental dynamics and the ecological functions of the eDNA and iDNA from the perspectives of environmental behavior, genetic information transmission, resistance to the environmental contaminants, and so on. By reviewing the status quo and the future vista of the e/iDNAs research, this article sheds light on exploring the ecological functioning of the e/iDNAs in the environmental microbiome.

Extracellular DNA refers to DNA fragments existing outside the cell, originating from various cell release mechanisms, including active secretion, cell lysis, and phage-mediated processes. Extracellular DNA serves as a vital environmental biomarker, playing crucial ecological and environmental roles in water bodies. This review is summarized the mechanisms of extracellular DNA release, including pathways involving cell lysis, extracellular vesicles, and type IV secretion systems. Then, the extraction and detection methods of extracellular DNA from water, soil, and biofilm are described and analyzed. Finally, we emphasize the role of extracellular DNA in microbial community systems, including its significant contributions to biofilm formation, biodiversity through horizontal gene transfer, and electron transfer processes. This review offers a comprehensive insight into the sources, distribution, functions, and impacts of extracellular DNA within aquatic environments, aiming to foster further exploration and understanding of extracellular DNA dynamics in aquatic environments as well as other environments.

Pseudomonas aeruginosa (P. aeruginosa) is a common pathogen associated with biofilm infections, which can lead to persistent infections. Therefore, there is an urgent need to develop new anti-biofilm drugs. DZ2002 is a reversible inhibitor that targets S-adenosylhomocysteine hydrolase and possesses anti-inflammatory and immune-regulatory activities. However, its anti-biofilm activity has not been reported yet.
Therefore, we investigated the effect of DZ2002 on P. aeruginosa PAO1 biofilm formation by crystal violet staining (CV), real-time quantitative polymerase chain reaction (RT-qPCR) and confocal laser scanning microscopy (CLSM). The results indicated that although DZ2002 didn\'t affect the growth of planktonic PAO1, it could significantly inhibit the formation of mature biofilms. During the inhibition of biofilm formation by DZ2002, there was a parallel decrease in the synthesis of alginate and the expression level of alginate genes, along with a weakening of swarming motility. However, these results were unrelated to the expression of lasI, lasR, rhII, rhIR. Additionally, we also found that after treatment with DZ2002, the biofilms and extracellular DNA content of PAO1 were significantly reduced. Molecular docking results further confirmed that DZ2002 had a strong binding affinity with the active site of S-adenosylhomocysteine hydrolase (SahH) of PAO1.
In summary, our results indicated that DZ2002 may interact with SahH in PAO1, inhibiting the formation of mature biofilms by downregulating alginate synthesis, extracellular DNA production and swarming motility. These findings demonstrate the potential value of DZ2002 in treating biofilm infections associated with P. aeruginosa.

Antibiotic resistance gene (ARG) transmission poses significant threats to human health. The effluent of wastewater treatment plants is demonstrated as a hotspot source of ARGs released into the environment. In this study, a synthetic microbiome containing nuclease-producing Deinococcus radiodurans was constructed to remove extracellular ARGs. Results of quantitative polymerase chain reaction (qPCR) showed significant reduction in plasmid RP4-associated ARGs (by more than 3 orders of magnitude) and reduction of indigenous ARG sul1 and mobile genetic element (MGE) intl1 (by more than 1 order of magnitude) in the synthetic microbiome compared to the control without D. radiodurans. Metagenomic analysis revealed a decrease in ARG and MGE diversity in extracellular DNA (eDNA) of the treated group. Notably, whereas eight antibiotic-resistant plasmids with mobility risk were detected in the control, only one was detected in the synthetic microbiome. The abundance of the nuclease encoding gene exeM, quantified by qPCR, indicated its enrichment in the synthetic microbiome, which ensures stable eDNA degradation even when D. radiodurans decreased. Moreover, intracellular ARGs and MGEs and pathogenic ARG hosts in the river receiving treated effluent were lower than those in the river receiving untreated effluent. Overall, this study presents a new approach for removing extracellular ARGs and further reducing the risk of ARG transmission in receiving rivers.

Chromium (Cr) is a heavy metal with a high toxicity and pathogenicity. Microbial reduction is an effective strategy to remove Cr(VI) at contaminated sites but suffers from the low populations and activities of Cr-reducing microorganisms in soils. This study proposed an in situ sonoporation-mediated gene transfer approach, which improved soil Cr(VI) reduction performance by delivering exogenous Cr-transporter chrA genes and Cr-reducing yieF genes into soil microorganisms with the aid of ultrasound. Besides the increasing populations of Cr-resistant bacteria and elevated copy numbers of chrA and yieF genes after sonoporation-mediated gene transfer, three new Cr-reducing strains were isolated, among which Comamonas aquatica was confirmed to obtain Cr-resistant capability. In addition, sonoporation-mediated gene transfer was the main driving force significantly shaping soil microbial communities owing to the predominance of Cr-resistant microbes. This study pioneered and evidenced that in situ soil sonoporation-mediated gene transfer could effectively deliver functional genes into soil indigenous microbes to facilitate microbial functions for enhanced bioremediation, e.g., Cr-reduction in this study, showing its feasibility as a chemically green and sustainable remediation strategy for heavy metal contaminated sites.

Neutrophils are a major subset of leukocytes in human circulating blood. In some circumstances, neutrophils release neutrophil extracellular traps (NETs). lnitially, NETs were considered to have a strong antibacterial capacity. However, currently, NETs have been shown to have a pivotal impact on various diseases. Different stimulators induce the production of different types of NETs, and their biological functions and modes of clearance do not appear to be the same. In this review, we will discuss several important issues related to NETs in order to better understand the relationship between NETs and diseases, as well as how to utilize the characteristics of NETs for disease treatment.

Microplastic (MP) biofilms provide a specific microniche for microbial life and are a potential hotspot for the horizontal gene transfer (HGT) of antibiotic resistance genes (ARGs). Nevertheless, the acquisition of ARGs in MP biofilms via natural transformation mediated by extracellular DNA (eDNA) has been rarely explored. This study demonstrated that MP biofilms promoted the natural transformation of extracellular ARGs at the single-cell and multi-species levels, compared to natural substrate (NS) biofilms and bacterioplankton. The transformation frequency on MP biofilms was up to 1000-fold compare to that on NS. The small MPs and aged MPs enhanced the ARG transformation frequencies up to 77.16-fold and 32.05-fold, respectively, compared with the large MPs and pristine MPs. The transformation frequencies on MP biofilms were significantly positively correlated with the bacterial density and extracellular polymeric substance (EPS) content (P < 0.05). Furthermore, MPs significantly increased the expression of the biofilm formation related genes (motA and pgaA) and DNA uptake related genes (pilX and comA) compared to NS and bacterioplankton. The more transformants colonized on MPs contributed to the enhanced transformation frequencies at the community-wide level. Overall, eDNA-mediated transformation in MP biofilms may be an important path of ARG spread, which was promoted by heterogeneous biofilm.

While the multiple functions of extracellular DNA (exDNA) in biofilm formation and electron transfer have been extensively studied in pure culture, its role in mixed anodic biofilm was still unknown. In this study, we employed DNase I enzyme to digest exDNA, thereby investigating its role in anodic biofilm formation based on the performance of four microbial electrolysis cells (MECs) groups with different DNase I enzyme concentration (0, 0.05, 0.1, 0.5 mg/mL). The responding time to reach 60 % maximum current of treatment group with DNase I enzyme has been significantly reduced to 83 %-86 % of the blank group (t-test, p < 0.01), indicating the exDNA digestion could promote the biofilm formation at the early stage. The anodic coulombic efficiency was enhanced by 10.74- 54.42 % in treatment group (t-test, p < 0.05), which could be ascribed to the higher absolute abundance of exoelectrogens. The lower relative abundance of exoelectrogens indicated the DNase I enzyme addition was beneficial for the enrichment of extensive species rather than exoelectrogens. As the DNase I enzyme augments the fluorescence signal of exDNA distribution in the small molecular weight region, implying the short chain exDNA could contribute to the biomass enhancement via boosting the most species enrichment. Furthermore, the exDNA alteration improved the complexity of microbial network. Our findings provide a new insight into the role of exDNA in the extracellular matrix of anodic biofilms.

Anaerobic ammonium oxidation (anammox) has been widely used for the sustainable removal of nitrogen from wastewater. Extracellular DNA (exDNA), as one of the main components of biofilms, not only determines the initial formation process, but also allows the three-dimensional structure to be maintained. Since the effects of exDNA on anammox biofilm formation are still poorly understood, this study elucidated the effects of exDNA on different stages of anammox biofilm establishment and maintenance under static conditions and its mechanism. The results revealed that exDNA mainly affected the maintenance stage of anammox biofilm formation. Compared with the absence of exDNA, nitrogen removal efficiency in the presence of exDNA was 6.17 % higher; the number of bacteria cells attached to the carrier was 2.23 times that in the absence of exDNA. The spatiotemporal distribution of bacteria was revealed by fluorescence in situ hybridization. After 30 days, the relative abundances of anammox in biofilms were 6.19 % and 0.4 % in the presence and absence of exDNA, respectively, indicating its positive role in anammox bacteria (AnAOB) adhesion and biofilm formation. The presence of exDNA in extracellular polymeric substances (EPS) promotes the synthesis of proteins and soluble microbial products. According to the extended Derjaguin-Landau-Verwey-Overbeek (X - DLVO) theory, the presence of exDNA also reduced the Lewis acid-base interaction energy and created favorable thermodynamic conditions for AnAOB adhesion. These findings advance our understanding of the role of exDNA in anammox-mediated biofilm formation and offer insights into the mechanism of exDNA in the establishment and maintenance stages.

Eradication of infectious biofilms is becoming increasingly difficult due to the growing number of antibiotic-resistant strains. This necessitates development of nonantibiotic-based, antimicrobial approaches. To this end, we designed a heterocatalytic metal-organic framework composed of zirconium 1,4-dicarboxybenzene (UiO-66) with immobilized Pt nanoparticles (Pt-NP/UiO-66). Pt-NP/UiO-66 enhanced singlet-oxygen generation compared with Pt nanoparticles or UiO-66, particularly in an acidic environment. Singlet-oxygen generation degraded phosphodiester bonds present in eDNA gluing biofilms together and therewith dispersed biofilms. Remaining biofilms possessed a more open structure. Concurrently, Pt-NP/UiO-66 stimulated macrophages to adapt a more M1-like, \"fighting\" phenotype, moving faster toward their target bacteria and showing increased bacterial killing. As a combined effect of biofilm dispersal and macrophage polarization, a subcutaneous Staphylococcus aureus biofilm in mice was more readily eradicated by Pt-NP/UiO-66 than by Pt nanoparticles or UiO-66. Therewith, heterocatalytic Pt-NP/UiO-66 metal-organic frameworks constitute a nonantibiotic-based strategy to weaken protective matrices and disperse infectious biofilms, while strengthening macrophages in bacterial killing.