NGS raw data 16SrDNA sequences are deposited in zenodo data bank: https://doi.org/10.5281/zenodo.8063253. | International audience | Highlights: • Severe marine plastic pollution impacts Southwest Indian Ocean insular ecosystems. • Plastic debris from Southwest Indian Ocean host rich microbiomes. • Proteobacteria dominate such marine plastic microbiomes. • These debris carry a consequent culturable bacterial flora including potential pathogens. • AMR bacteria hitchhike on these plastics.Abstract: The increasing threats to ecosystems and humans from marine plastic pollution require a comprehensive assessment. We present a plastisphere case study from Reunion Island, a remote oceanic island located in the Southwest Indian Ocean, polluted by plastics. We characterized the plastic pollution on the island's coastal waters, described the associated microbiome, explored viable bacterial flora and the presence of antimicrobial resistant (AMR) bacteria. Reunion Island faces plastic pollution with up to 10,000 items/km 2 in coastal water. These plastics host microbiomes dominated by Proteobacteria (80 %), including dominant genera such as Psychrobacter, Photobacterium, Pseudoalteromonas and Vibrio. Culturable microbiomes reach 10 7 CFU/g of microplastics, with dominance of Exiguobacterium and Pseudomonas. Plastics also carry AMR bacteria including β-lactam resistance. Thus, Southwest Indian Ocean islands are facing serious plastic pollution. This pollution requires vigilant monitoring as it harbors a plastisphere including AMR, that threatens pristine ecosystems and potentially human health through the marine food chain.

This study investigated the effects of adding biochar (BC) on the fate of ciprofloxacin (CIP) and its related antibiotic tolerance (AT) in activated sludge. Three activated sludge reactors were established with different types of BC, derived from apple, pear, and mulberry tree, respectively, and one reactor with no BC. All reactors were exposed to an environmentally relevant level of CIP that acted as a definitive selective pressure significantly promoting AT to four representative antibiotics (CIP, ampicillin, tetracycline, and polymyxin B) by up to two orders of magnitude. While CIP removal was negligible in the reactor without BC, the BC-dosed reactors effectively removed CIP (70–95% removals) through primarily adsorption by BC and biodegradation/biosorption by biomass. The AT in the BC-added reactors was suppressed by 10–99%, compared to that without BC. The BC addition played a key role in sequestering CIP, thereby decreasing the selective pressure that enabled the proactive prevention of AT increase. 16S rRNA gene sequencing analysis showed that the BC addition alleviated the CIP-mediated toxicity to community diversity and organisms related to phosphorous removal. Machine learning modeling with random forest and support vector models using AS microbiome data collectively pinpointed Achromobacter selected by CIP and strongly associated with the AT increase in activated sludge. The identification of Achromobacter as an important AT bacteria revealed by the machine learning modeling with multiple models was also validated with a linear Pearson's correlation analysis. Overall, our study highlighted Achromobacter as a potential useful sentinel for monitoring AT occurring in the environment and suggested BC as a promising additive in wastewater treatment to improve micropollutant removal, mitigate potential AT propagation, and maintain community diversity against toxic antibiotic loadings.

Heavy metal toxicity has become an impediment to agricultural productivity, which presents major human health concerns in terms of food safety. Among them, arsenic (As) a non-essential heavy metal has gained worldwide attention because of its noxious effects on agriculture and public health. The increasing rate of global warming and anthropogenic activities have promptly exacerbated As levels in the agricultural soil, thereby causing adverse effects to crop genetic and phenotypic traits and rendering them vulnerable to other stresses. Conventional breeding and transgenic approaches have been widely adapted for producing heavy metal resilient crops; however, they are time-consuming and labor-intensive. Hence, finding new mitigation strategies for As toxicity would be a game-changer for sustainable agriculture. One such promising approach is harnessing plant microbiome in the era of ‘omics’ which is gaining prominence in recent years. The use of plant microbiome and their cocktails to combat As metal toxicity has gained widespread attention, because of their ability to metabolize toxic elements and offer an array of perquisites to host plants such as increased nutrient availability, stress resilience, soil fertility, and yield. A comprehensive understanding of below-ground plant-microbiome interactions and their underlying molecular mechanisms in exhibiting resilience towards As toxicity will help in identifying elite microbial communities for As mitigation. In this review, we have discussed the effect of As, their accumulation, transportation, signaling, and detoxification in plants. We have also discussed the role of the plant microbiome in mitigating As toxicity which has become an intriguing research frontier in phytoremediation. This review also provides insights on the advancements in constructing the beneficial synthetic microbial communities (SynComs) using microbiome engineering that will facilitate the development of the most advanced As remedial tool kit in sustainable agriculture.

Biodegradable plastic mulch film (BDM) is an environmentally friendly alternative to conventional polyethylene mulch, and has been growingly used in agriculture. However, practical degradation performance of BDM, especially the widely used type of blended polylactic acid (PLA)/polybutylene adipate (PBAT) in different ratios, and microbial alteration in soil environments, remain largely unrevealed. In this study, four types of BDM blended with 40–80% PLA and 20–60% PBAT were comparatively investigated through microcosm soil incubation experiments for 105 days, and combined with conditions of different soil moisture or pH. Microbiome within film-surrounding soil were assayed using 16 S rRNA high-throughput sequencing. Results showed a trend of increasing degradation efficiency with the increase of PLA proportion, and 70% PLA and 30% PBAT group presented the highest weight loss rate, i.e., 60.16 ± 5.86%. In addition, degradation and aging of PLA/PBAT varied among different soil moisture and pH values. A moderate moisture, i.e., 60% and a neutral pH7.0 caused significantly high degradation efficiency compared to other moisture or pH conditions. Moreover, bacterial abundance and community structure in the surrounding soil were related to soil moisture and pH. PLA/PBAT incubation treatment induced a remarkable increase in abundance of degradation-related species Pseudomonas and Sphingomonas. Bacterial richness and diversity in soil correspondingly respond to ratio-different PLA/PBAT's degradation under moisture/pH-different conditions through a redundancy analysis. Altogether, these findings indicate that practical degradation of PLA/PBAT film is closely related to soil environments and bacterial community. It is significant for the application of biodegradable plastics in agriculture on the perspective of soil sustainability.

Whalers Bay, in Deception Island, has one of the most anthropogenically impacted areas in Maritime Antarctica. However, considering the volcanic nature (high concentrations of heavy metals) of Deception Island's soils, this putative anthropogenic impact should be carefully investigated. In this context, the objective of this study was to compare resistome profiles of impacted and non-impacted areas in Deception Island (Whalers Bay, Crater Lake, and Fumarole Bay) and Livingston Island (Hannah Point) in order to investigate the microbiome tolerance/resistance mechanisms selected as a function of environmental drivers. Metagenomics was used to search for genes conferring resistance/tolerance to antibiotics, biocides, and heavy metals. Whalers Bay has a greater diversity of antibiotic, biocide, and heavy metal resistance classes found in its microbiomes. However, Hannah Point, at Livingston Island, has a greater abundance of antibiotic and biocide resistance/tolerance genes. The microbiome of Deception Island's non-impacted areas (Crater Lake and Fumarole Bay) showed resistance/tolerance genes almost entirely to heavy metals. Pb was found in higher concentrations in Whalers Bay soil in comparison to the other areas, indicating human contamination. The non-metric multidimensional scaling (NMDS) analysis revealed that Pb concentrations influenced resistome profiles in Whalers Bay soil. Despite the effect of Pb on the microbial communities of Whalers Bay, most heavy metal concentrations did not have a significant impact on resistome genes, suggesting that the volcanic soil heavy metal concentration of Deception Island has little biological influence.

Various microplastics (MPs) are found in the environment and organisms. MP residues in organisms can affect health; however, their impacts on metabolism in aquatic organisms remain unclear. In this study, zebrafish embryos were exposed to polyethylene MPs with sizes ranging from 1 to 4 μm at concentrations of 0, 10, 100, and 1000 μg/L for 7 days. Through qPCR technology, the results indicated that zebrafish exposed to polyethylene MPs exhibited significant change in microbes of the phyla Firmicutes, Bacteroidetes, Proteobacteria, and Verrucomicrobia, etc. Moreover, 16S RNA gene sequencing revealed that there was a significant difference in alpha diversity between the control and 1000 μg/L MP-treated groups. At the genus level, the abundance of Aeromonas, Shewanella, Microbacterium, Nevskia and Methyloversatilis have increased remarkably. Conversely, the abundance of Pseudomonas, Ralstonia and Stenotrophomonas were significant reduction after MPs exposure. In addition, the levels of TG (triglyceride), TCHO (total cholesterol), NEFA (nonesterified fatty acid), TBA (total bile acid), GLU (glucose) and pyruvic acid significantly changed in MP-treated larval zebrafish, indicating that their metabolism was disturbed by MPs. Transcriptional levels of glucose and lipid metabolism-related genes showed a decreasing trend. Furthermore, LC/MS-based nontargeted metabolomics analysis demonstrated that a total of 59 phospholipid-related substances exhibited significant changes in larval fish treated with 1000 μg/L MPs. The mRNA levels of phospholipid metabolism-related genes were also obviously changed. Pearson correlation analysis indicated that the abundance of Aeromonas, Shewanella and Chitinibacter bacteria showed a negative correlation with most phospholipids, while Nevskia, Parvibacter and Lysobacter showed a positive correlation with most phospholipids. Based on these results, it is suggested that 1–4 μm PE-MPs could impact the microbiome and metabolism of larval zebrafish. All of these results indicated that the health risk of MPs cannot be ignored.

Rhizobia and arbuscular mycorrhiza fungi (AMF) are important symbiotic microbes that are advantageous to plants growing in metal-contaminated soil. However, it remains unclear how inoculated microbes affect rhizosphere microbial communities or whether subsequent changes in rhizosphere microbiomes contribute to improving plant resistance under metal stress. This study investigated the effects of rhizobia and AMF inoculation on alfalfa resistance to Cd stress. The response of rhizosphere microbial communities to inoculation and its role in increasing alfalfa’ ability to cope with stress were further analyzed using high-throughput sequencing of 16S and ITS rRNA genes. Results showed that single rhizobia or AMF inoculation significantly improved alfalfa resistance to Cd stress, while their co-inoculation resulted in the greatest overall improvement. Improved resistance was reflected by the significant mitigation of Cd-induced lipid peroxidation and reactive oxygen species (ROS) stress caused by increases in antioxidant enzyme activities along with co-inoculation. Furthermore, co-inoculation significantly altered the rhizosphere microbial community structure by decreasing fungal community diversity and increasing bacterial community diversity. Results of partial least squares path modeling (PLS-PM) and variation partitioning analysis (VPA) showed that the rhizosphere bacterial community predominated over the fungal community with respected to improvements in resistance to Cd stress under the co-inoculation treatments. This improvement was specifically seen in the enrichment of certain key bacterial taxa (including Proteobacteria, Actinobacteria, Acidobacteria, and Chloroflexi) induced by the rhizobia and AMF co-inoculation, enhancing alfalfa’ ability to uptake rhizosphere nutrients and reduce its release of photosynthetically-derived carbon (C) into soil. Our findings revealed that the co-inoculation of multiple symbiotic microbes can assist plants to effectively cope with Cd stress, providing a greater understanding of rhizosphere bacterial taxa in the microbe-induced phytomanagement.

Cyanobacterial harmful algal blooms (CyanoHABs) have been found to transmit from N₂ fixer-dominated to non-N₂ fixer-dominated in many freshwater environments when the supply of N decreases. To elucidate the mechanisms underlying such “counter-intuitive” CyanoHAB species succession, metatranscriptomes (biotic data) and water quality-related variables (abiotic data) were analyzed weekly during a bloom season in Harsha Lake, a multipurpose lake that serves as a drinking water source and recreational ground. Our results showed that CyanoHABs in Harsha Lake started with N₂-fixing Anabaena in June (ANA stage) when N was high, and transitioned to non-N₂-fixing Microcystis- and Planktothrix-dominated in July (MIC-PLA stage) when N became limited (low TN/TP). Meanwhile, the concentrations of cyanotoxins, i.e., microcystins were significantly higher in the MIC-PLA stage. Water quality results revealed that N species (i.e., TN, TN/TP) and water temperature were significantly correlated with cyanobacterial biomass. Expression levels of several C- and N-processing-related cyanobacterial genes were highly predictive of the biomass of their species. More importantly, the biomasses of Microcystis and Planktothrix were also significantly associated with expressions of microbial genes (mostly from heterotrophic bacteria) related to processing organic substrates (alkaline phosphatase, peptidase, carbohydrate-active enzymes) and cyanophage genes. Collectively, our results suggest that besides environmental conditions and inherent traits of specific cyanobacterial species, the development and succession of CyanoHABs are regulated by co-occurring microorganisms. Specifically, the co-occurring microorganisms can alleviate the nutrient limitation of cyanobacteria by remineralizing organic compounds.

Animals living in urban river systems play critical roles in the dissemination of microbiome and antibiotic resistance that poses a strong threat to public health. This study provides a comprehensive profile of microbiota and antibiotic resistance genes (ARGs) of sharpbelly (Hemiculter leucisculus) and the surrounding water from five sites along the Ba River. Results showed Proteobacteria, Firmicutes and Fusobacteria were the dominant bacteria in gut of H. leucisculus. With the aggravation of water pollution, bacterial biomass of fish gut significantly decreased and the proportion of Proteobacteria increased to become the most dominant phylum eventually. To quantify the contributions of influential factors on patterns of gut microbiome with structural equation model (SEM), water bacteria were confirmed to be the most stressors to perturb fish gut microbiome. SourceTracker model indicated that deteriorating living surroundings facilitated the invasion of water pathogens to fish gut eco-environments. Additionally, H. leucisculus gut is an important reservoir of ARGs in Ba River with relative abundance up to 9.86 × 10⁻¹/copies. Among the ARGs, tetracycline and quinolone resistance genes were detected in dominant abundance. Deterioration of external environments elicited the accumulation of ARGs in fish gut. Intestinal class I integron, environmental heavy metal residues and gut bacteria were identified as key drivers of intestinal ARGs profiles in H. leucisculus. Analysis of SEM and co-occurrence patterns between ARGs and bacterial hosts indicated that class I integron and bacterial community played vital roles in ARGs transmission through water-fish pathway. In general, this study highlighted hazards of water contamination to microbiome and ARGs in aquatic animals and provided a new perspective to better understand the bacteria and ARGs dissemination in urban river ecosystems.

Dietary exposure to chemicals alters the diversity of microbiome communities and can lead to pathophysiological changes in the gastrointestinal system. The organochlorine pesticide dieldrin is a persistent environmental contaminant that bioaccumulates in fatty tissue of aquatic organisms. The objectives of this study were to determine whether environmentally-relevant doses of dieldrin altered gastrointestinal morphology and the microbiome of zebrafish. Adult zebrafish at ∼4 months of age were fed a measured amount of feed containing either a solvent control or one of two doses of dieldrin (measured at 16, and 163.5 ng/g dry weight) for 4 months. Dieldrin body burden levels in zebrafish after four-month exposure were 0 (control), 11.47 ± 1.13 ng/g (low dose) and 18.32 ± 1.32 ng/g (high dose) wet weight [mean ± std]. Extensive histopathology at the whole organism level revealed that dieldrin exposure did not induce notable tissue pathology, including the gastrointestinal tract. A repeated measure mixed model analysis revealed that, while fish gained weight over time, there were no dieldrin-specific effects on body weight. Fecal content was collected from the gastrointestinal tract of males and 16S rRNA gene sequencing conducted. Dieldrin at a measured feed dose of 16 ng/g reduced the abundance of Firmicutes, a phylum involved in energy resorption. At the level of class, there was a decrease in abundance of Clostridia and Betaproteobacteria, and an increase in Verrucomicrobiae species. We used a computational approach called predicted relative metabolomic turnover (PRMT) to predict how a shift in microbial community composition affects exchange of metabolites. Dieldrin was predicted to affect metabolic turnover of uroporphyrinogen I and coproporphyrinogen I [enzyme]−cysteine, hydrogen selenide, selenite, and methyl-selenic acid in the fish gastrointestinal system. These pathways are related to bacterial heme biosynthesis and selenium metabolism. Our study demonstrates that dietary exposures to dieldrin can alter microbiota composition over 4 months, however the long-term consequences of such impacts are not well understood.