Unraveling the effect of micro/nanoplastics on the occurrence and horizontal transfer of environmental antibiotic resistance genes: Advances, mechanisms and future prospects.

Microplastics can not only serve as vectors of antibiotic resistance genes (ARGs), but also they and even nanoplastics potentially affect the occurrence of ARGs in indigenous environmental microorganisms, which have aroused great concern for the development of antibiotic resistance. This article specifically reviews the effects of micro/nanoplastics (concentration, size, exposure time, chemical additives) and their interactions with other pollutants on environmental ARGs dissemination. The changes of horizontal genes transfer (HGT, i.e., conjugation, transformation and transduction) of ARGs caused by micro/nanoplastics were also summarized. Further, this review systematically sums up the mechanisms of micro/nanoplastics regulating HGT process of ARGs, including reactive oxygen species production, cell membrane permeability, transfer-related genes expression, extracellular polymeric substances production, and ARG donor-recipient adsorption/contaminants adsorption/biofilm formation. The underlying mechanisms in changes of bacterial communities induced by micro/nanoplastics were also discussed as it was an important factor for structuring the profile of ARGs in the actual environment, including causing environmental stress, providing carbon sources, forming biofilms, affecting pollutants distribution and environmental factors. This review contributes to a systematical understanding of the potential risks of antibiotic resistance dissemination caused by micro/nanoplastics and provokes thinking about perspectives for future research and the management of micro/nanoplastics and plastics.

Diverse bacterial hosts and potential risk of antibiotic resistomes in ship ballast water revealed by metagenomic binning.

Ship ballast water promoting the long-range migration of antibiotic resistance genes (ARGs) has raised a great concern. This study attempted to reveal ARGs profile in ballast water and decipher their hosts and potential risk using metagenomic approaches. In total, 710 subtypes across 26 ARG types were identified among the ballast water samples from 13 ships of 11 countries and regions, and multidrug resistance genes were the most dominant ARGs. The composition of ARGs were obviously different across samples, and only 5% of the ARG subtypes were shared by all samples. Procrustes analysis showed the bacterial community contributed more than the mobile genetic elements (MGEs) in shaping the antibiotic resistome. Further, 79 metagenome-assembled genomes (46 genera belong to four phyla) were identified as ARG hosts, with predominantly affiliated with the Proteobacteria. Notably, potential human pathogens (Alcaligenes, Mycolicibacterium, Rhodococcus and Pseudomonas) were also recognized as the ARG hosts. Above 30% of the ARGs hosts contained the MGEs simultaneously, supporting a pronounced horizontal gene transfer capability. A total of 43 subtypes (six percent of overall ARGs) of ARGs were assessed with high-risk, of which 23 subtypes belonged to risk Rank I (including rsmA, ugd, etc.) and 20 subtypes to the risk Rank II (including aac(6)-I, sul1, etc.). In addition, antibiotic resistance risk index indicated the risk of ARGs in ballast water from choke points of maritime trade routes was significantly higher than that from other regions. Overall, this study offers insights for risk evaluation and management of antibiotic resistance in ballast water.

Exploring the bacterial diversity and composition with special emphasis on pathogens in ship ballast water and sediments using full-length 16S rRNA gene sequencing.

Accurate detecting bacterial communities in ballast water and sediments supports risk management. This study uses full-length 16S rRNA gene sequencing to investigate the bacterial communities in ballast water and sediments, focusing on detecting pathogens. The results indicate that full-length sequencing more accurately reveals the species diversity. There is a significant difference (P < 0.05) in bacterial communities between ballast water and sediments, despite both being dominated by the Proteobacteria phylum. Thirty human and fish pathogens were identified by full-length sequencing, yet only five pathogens were detected from V3-V4 sequencing. Notably, emerging pathogens such as Citrobacter freundii and Nocardia nova are detected in samples, which are harmful to aquaculture and human health. Several opportunistic pathogens were also identified. In summary, this study provides important insights into the bacterial communities in ballast water and sediments, highlighting the need for strict management.

Responses of compositions, functions, and assembly processes of bacterial and microeukaryotic communities to long-range voyages in simulated ballast water.

Ballast water is one of the main vectors for the spread of harmful organisms among geologically isolated waters. However, the successional processes of microbial functions and assembly processes in ballast water during the long-term shipping voyage remain unclear. In this study, the compositions, ecological functions, community assembly, and potential environmental drivers of bacteria and microeukaryotes were investigated in simulated ballast water microcosms for 120 days. The results showed that the diversity and compositions of the bacterial and microeukaryotic communities varied significantly in the initial 40 days (T0∼T40 samples) and then gradually converged. The relative abundance of Proteobacteria showed a distinct tendency to decrease (87.90%-41.44%), while that of Ascomycota exhibited an increasing trend (6.35%-62.12%). The functional groups also varied significantly over time and could be related to the variations of the microbial community. The chemoheterotrophy and aerobic chemoheterotrophy functional groups for bacteria decreased from 44.80% to 28.02% and from 43.77% to 25.39%, respectively. Additionally, co-occurrence network analysis showed that the structures of the bacterial community in T60∼T120 samples were more stable than those in T0∼T40 samples. Stochastic processes also significantly affected the community assembly of bacteria and microeukaryotes. pH played the most significant role in driving the structures and assembly processes of the bacterial and microeukaryotic communities. The results of this study could aid in the understanding of variations in the functions and ecological processes of bacterial and microeukaryotic communities in ballast water over time and provide a theoretical basis for its management.

Insight into the responses of antibiotic resistance genes in microplastic biofilms to zinc oxide nanoparticles and zinc ions pressures in landfill leachate.

Microplastic (MP) biofilms are hotspots of antibiotic resistance genes (ARGs) in landfill environment. MP biofilms in landfill leachate coexist with heavy metals and metallic nanoparticles (NPs) that considered to be the selective agents of ARGs. However, the effects of these selective pressures on ARGs in MP biofilms and their differences in MP-surrounding leachate have not been well understood. Herein, the changes of ARG abundances in MP biofilms and corresponding leachate under zinc oxide (ZnO) NPs and zinc ion (Zn2+) pressures were comparatively analyzed. The presence of ZnO NPs and Zn2+ promoted the enrichment of ARGs in MP biofilms, and the enrichment was more pronounced in ZnO NPs groups. ZnO NPs and especially Zn2+ mainly decreased the abundances of ARGs in leachate. The increase of integron abundances and reactive oxygen species production in MP biofilms implied the enhanced potential for horizontal transfer of ARGs under ZnO NPs and Zn2+ pressures. Meanwhile, the co-occurrence pattern between ARGs and bacterial genera in MP biofilms with more diverse potential ARG hosts was more complex than in leachate, and the enrichment of ARG-hosting bacteria in MP biofilms under ZnO NPs and Zn2+ pressures supported the enrichment of ARGs.

The prevalence of potential pathogens in ballast water and sediments of oceangoing vessels and implications for management.

Ballast water and sediments can serve as prominent vectors for the widespread dispersal of pathogens between geographically distant areas. However, information regarding the diversity and distribution of the bacterial pathogens in ballast water and sediments is highly limited. In this study, using high-throughput sequencing and quantitative PCR, we investigated the composition and abundance of potential pathogens, and their associations with indicator microorganisms. We accordingly detected 48 potential bacterial pathogens in the assessed ballast water and sediments, among which there were significant differences in the compositions and abundances of pathogenic bacterial communities characterizing ballast water and sediments. Rhodococcus erythropolis, Bacteroides vulgatus, and Vibrio campbellii were identified as predominant pathogens in ballast water, whereas Pseudomonas stutzeri, Mycobacterium paragordonae, and Bacillus anthracis predominated in ballast sediments. Bacteroidetes, Vibrio alginolyticus, Vibrio parahaemolyticus, and Escherichia coli were generally detected with median values of 8.54 × 103-1.22 × 107 gene copies (GC)/100 mL and 1.16 × 107-3.97 × 109 GC/100 g in ballast water and sediments, respectively. Notably, the concentrations of Shigella sp., Staphylococcus aureus, and V. alginolyticus were significantly higher in ballast sediments than in the water. In addition, our findings tend to confirm that the indicator species specified by the International Maritime Organization (IMO) might underestimate the pathogen risk in the ballast water and sediments, as these bacteria were unable to predict some potential pathogens assessed in this study. In summary, this study provides a comprehensive insight into the spectrum of the potential pathogens that transferred by ship ballast tanks and emphasizes the need for the implementation of IMO convention on ballast sediment management.

Deciphering the characterization, ecological function and assembly processes of bacterial communities in ship ballast water and sediments.

Various microorganisms are transported worldwide via the water and sediments inside ship ballast tanks. Nevertheless, the ecological functions and assembly processes of bacterial communities in ballast water and sediments remain poorly understood. Here, we investigated the bacterial composition, community assembly processes, and putative functions through analyses of 70 ballast water and sediment samples obtained from various ships. The results showed that the ballast sediments contained a higher diversity of bacterial communities, whereas the ballast water was characterized by the dominance of Proteobacteria. Both the composition and potential function structures of bacterial communities were clearly different between the ballast water and sediment samples. The ballast water exhibited an abundance of microorganisms that involved in sulfur oxidation, whereas the bacterial species associated with nitrogen metabolism were abundant in the sediments. Co-occurrence network analysis revealed that the communities in ballast sediment samples possessed more complex network structures with higher modularity and positive associations among bacterial populations. Stochastic processes, especially the dispersal limitation process played the most important influence in the assembly of the communities in ballast water. Meanwhile, the bacterial communities in the ballast sediments were primarily governed by the homogeneous selection of determinacy. The results from this study will help us understand the ecological processes related to the bacterial communities in the ballast tanks and provide a foundation for the management of ballast water and sediments.

Transfer of antibiotic-resistant bacteria via ballast water with a special focus on multiple antibiotic-resistant bacteria: A survey from an inland port in the Yangtze River.

Ship ballast water can transfer harmful organisms, including antibiotic-resistant bacteria (ARB), among geographically isolated waters. In this study, the presence and composition of ARB and multiple ARB (MARB) were investigated in the ballast waters of 30 vessels sailing to the Port of Jiangyin (Jiangsu Province, China). ARB were detected in 83.3% of the ship's ballast water samples. Moreover, penicillin- and cephalothin-resistant bacteria were the most and least prevalent ARB in the ballast waters, respectively. Oxytetracycline-, chloramphenicol-, tetracycline-, and vancomycin-resistant bacteria were also detected at high concentrations. The multiple antibiotic resistance index demonstrated the presence of MARB, which exceeded 200% in the ballast waters of five ships. Furthermore, 15 species, including the human opportunistic pathogens Vibrio alginolyticus and Serratia nematodiphila, were resistant to at least three antibiotics. Therefore, the potential ecological risk of ARB warrants further attention because of their effective invasion by ballast water.

Elucidating the role of earthworms in N2O emission and production pathway during vermicomposting of sewage sludge and rice straw.

Vermicomposting is a sustainable option for the recycling of biodegradable organic waste. However, it also produces nitrous oxide (N2O), which is a highly potent greenhouse gas. In this study, the N2O stable isotope and functional genes for nitrogen cycling were determined to investigate the sources of N2O during vermicomposting. The results showed that vermicomposting promoted the organic degradation and nitrogen nitrification, and the presence of earthworms increased the emission of N2O during vermicomposting compared to that during the control treatment with no earthworms. The site preference analysis of N2O stable isotope showed that both nitrification and denitrification were present during the early stages of vermicomposting, while nitrification was the dominant contributor to N2O production in the later stages. Moreover, earthworms increased the gene copies of amoA, and stimulated the nitrifying bacteria, and hence, increased the N2O emission via nitrification. In addition, the activity of earthworms reduced the gene number of nosZ during vermicomposting, while the denitrification was the main source of N2O in the earthworm gut, as the conditions inside the gut inhibited nosZ. Overall, nitrification was the major pathway (55.8-88.7 %) for N2O production, which was promoted by the introduction of earthworms through nitrification.

Vessel transport of antibiotic resistance genes across oceans and its implications for ballast water management.

The emergence and spread of antibiotic resistance are major threats to ecosystems and human health. Transoceanic channels (e.g., ship ballast water) can transfer harmful aquatic organisms across geographically isolated waters. However, the occurrence of antibiotic resistance genes (ARGs) in ship ballast water and their relationship with microbial communities and environmental factors remain unknown. In this study, ballast water from 28 vessels sailing to Shanghai and Jiangyin (China) were collected, and the ARGs in these water samples were investigated. Considerable levels of ARGs and integrase of the class-I integrons (intI1) were detected in all ballast water samples. sul1 and tetQ were the most and least abundant ARGs in ballast water samples, respectively. The ARGs were strongly correlated with those of the 16S rRNA and intI1 genes. Ballast water exchange can reduce the absolute abundance of some kinds of ARGs while increasing the relative abundance of several ARGs (e.g., mefA, mexF, strB, sul1, and tetQ). Moreover, the bacterial hosts of ARGs were generally different in the unexchanged ballast water (UEBW) and exchanged ballast water (EBW). In particular, Leisingera and unclassified_Erythrobacteraceae were the main ARGs-associated genera in the EBW, while Pseudohongiella, Cycloclasticus, OM43_clade, norank_f_Rhodospirillaceae, and norank_f_Rhodobacteraceae were the dominant ARGs hosts in the UEBW. Overall, ship ballast water is an effective moving carrier for the global transference of ARGs, and its sufficient management is required for mitigating ARGs propagation across oceans.

Effects of holding time on the diversity and composition of potential pathogenic bacteria in ship ballast water.

Ballast water is a common vector for the transport of invasive species to new marine and aquatic environments. We used a metagenomics approach to examine the diversity and composition of potential pathogens communities in ballast water from ships in the route of China- Southeast Asia (CSEA). 16 kinds of potential pathogenic genus were detected in the ballast water. Interestingly, the ballast holding time had an important effect on the distribution of potential pathogens in ballast water. The abundance of Pseudoalteromonas in the longer ballast water holding time was less than the shorter ballast water holding time. Bacteroides had completely disappeared in the long ballast holding time samples. Moreover, The Shannon index of samples with longer ballast water holding time (1.80 ± 0.07) was higher than those with shorter ballast water holding time (0.83 ± 0.13). The potential pathogenic genus (Arcobacter, Aeromonas, Enterobacter and so on) lived in the long ballast holding time vessels had more diversity. Besides, the total suspended solids (TSS), total organic carbon (TOC) and particulate organic carbon (POC) had a strong positive correlation with most potential pathogens in the ballast water, while the dissolved oxygen (DO) had a clear negative correlation with the potential pathogens in the longer ballast holding time samples. In conclusion, these results provide detailed descriptions of the characteristics of the potential pathogens present in ballast water, document significant potential pathogens diversity, and indicate the importance of ballast holding time for potential pathogens lived in ballast water.

Effects of earthworms on nitrogen transformation and the correspond genes (amoA and nirS) in vermicomposting of sewage sludge and rice straw.

The effects of earthworms on nitrogen transformation and the responsible functional genes during disposal of sewage sludge and rice straw were investigated in this study. Vermicomposting resulted in the lower pH and total organic carbon (TOC) compared to the control treatment without earthworms. Moreover, the presence of earthworms could promote the nitrogen mineralization and nitrification process in vermicomposting. Earthworms increased the activity of ammonia monooxygenase and abundance of amoA-nitrifier and reduced its diversity, whereas they reduced the density of nirS-denitrifying bacteria but enhanced its diversity. Nitrosospira was the dominant amoA-nitrifier and earthworms stimulated its growth in the vermicomposting. The presence of earthworms could also affect the community composition of nirS-denitrifying bacteria despite most of the nirS-denitrifier was not be classified at the genus level. In conclusion, the presence of earthworms had significant influence on the diversity and abundances of amoA and nirS genes and affect the nitrogen bio-transformation in vermicomposting.

Effects of C/N ratio and earthworms on greenhouse gas emissions during vermicomposting of sewage sludge.

The emissions of greenhouse gases (CO2, CH4, and N2O) during bio-stabilization of sewage sludge under different C/N ratios with/without Eisenia fetida were evaluated in this study. Vermicomposting led to the more significant reductions of pH, TOC and C/N ratio compared to the control treatment without earthworms. C/N ratio had a significant effect on the emission of N2O, whereas its influences on CO2 or CH4 emission were not obvious. Earthworms reduced the CH4 emission greatly, although the CO2 emission was not affected by earthworms. Furthermore, a higher emission of N2O was observed in vermicomposting compared to the control. Both the C/N ratios and earthworms exerted statistically significant effects on the total GHG emission. Vermicomposting could enhance the degradation of organics and accelerate the mineralization process of nitrogen, whereas the increase in the C/N ratio could reduce GHG emission during vermicomposting of sewage sludge.

Abundances and profiles of antibiotic resistance genes as well as co-occurrences with human bacterial pathogens in ship ballast tank sediments from a shipyard in Jiangsu Province, China.

Ship ballasting operations may transfer harmful aquatic organisms across global ocean. This study aims to reveal the occurrences and abundances of antibiotic resistance genes (ARGs) and human bacterial pathogens (HBPs) in ballast tank sediments. Nine samples were collected and respectively analyzed by real-time quantitative PCR and high-throughput sequencing technologies. Ten ARGs (aadA1, blaCTX-M, blaTEM, ermB, mefA, strB, sul1, sul2, tetM, and tetQ) and the Class-I integron gene (intI1) were highly prevalent (105-109 gene copies/g) in ballast tank sediments. The sul1 was the most abundant ARG with the concentration of 108-109 copies/g and intI1 was much more abundant than the ARGs in ballast tank sediments. The strong positive correlations between intI1 and ARGs (blaCTX-M, sul1, sul2 and tetM) indicated the potential spread of ARGs via horizontal gene transfer. In ballast tank sediments, 44 bacterial species were identified as HBPs and accounted for 0.13-21.46% of the total bacterial population although the three indicator pathogenic microbes (Vibrio cholerae, Escherichia coli, and Enterococci) proposed by the International Maritime Organization were not detected. Pseudomonas pseudoalcaligenes, Enterococcus hirae, Shigella sonnei and Bacillus anthracis were the dominant pathogens in ballast tank sediments. Zn and P in sediments had positive effects on the ARGs. Network analysis results indicated that sul1 and sul2 genes existed in several bacterial pathogens. Ballast tank sediments could be regarded as a carrier for the migration of ARGs. It is important to manage ballast tank sediments reasonably in order to prevent the dissemination of ARGs and bacterial pathogens.

Exploring the effects of earthworms on bacterial profiles during vermicomposting process of sewage sludge and cattle dung with high-throughput sequencing.

This study aims to reveal the effects of earthworms (Eisenia fetida) on bacterial profiles during the vermicomposting process of sewage sludge and cattle dung with the high-throughput sequencing technology. The earthworms could accelerate organic degradation and improve the stabilization process. Moreover, the addition of earthworms not only affected the bacterial numbers, but also increased the bacterial community diversity. The activity of earthworms had significant effects on the bacterial community structure as the bacterial community was clearly different between the vermicomposting and the control treatment. Furthermore, the earthworms affected the physical and chemical properties of substrates, thus promoting the growth of some microorganisms, such as Flavobacteria, Acidbacteria, and Planctomycetes. Earthworms largely inhibited the growth of various human pathogenic bacteria. In summary, earthworms significantly affected the bacterial community in vermicomposting and it could be applied as an authentically effective technique for the stabilization of organic wastes.

Composition and influencing factors of bacterial communities in ballast tank sediments: Implications for ballast water and sediment management.

This study aims to reveal the composition and influencing factors of bacterial communities in ballast tank sediments. Nine samples were collected and their 16S rRNA gene sequences were analyzed by high-throughput sequencing. The analysis results showed the Shannon index in ballast tank sediments was in the range of 5.27-6.35, which was significantly higher than that in ballast water. Acidobacteria, Actinobacteria, Bacteroidetes, Chloroflexi and Proteobacteria were the dominant phyla and accounted for approximately 80% of all 16S rRNA gene sequences of the samples. Besides, the high contents of sulfate reducing bacteria (SRB) and sulfur oxidizing bacteria were detected in sediments, indicating that the corrosion of metal caused by SRB might occur in ballast tank. In addition, the trace of human fecal bacteria and candidate pathogens were also detected in ballast tank sediments, and these undesirable microbes reduced the effect of ballast water exchange. Furthermore, C and N had significant effects on the bacterial community composition in ballast tank sediments. In conclusion, our findings suggest that the proper management and disposal of the ballast tank sediments should be considered in order to reduce the negative impact and ecological risks related to ballast water and sediments.

Occurrences, composition profiles and source identifications of polycyclic aromatic hydrocarbons (PAHs), polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) in ship ballast sediments.

The aim of this study was to investigate the levels of persistent organic pollutants (POPs) including polycyclic aromatic hydrocarbons (PAHs), polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) in ship ballast sediments. The ballast sediment samples were collected from six merchant ships docked in 2015 in Jiangyin City, China. Ballast sediments represent a potential vector for the transport of POPs and invasive species between marine environments. An attempt was also made to determine the sources of these compounds in the ballast sediment. The results indicated ballast sediments generally contain greater amounts of BDE-209 and comparable amounts of PAHs, PBDEs (exclusive of BDE-209) and PCBs compared to those in marine surface sediments. Based on the sediment quality guidelines, PAHs and PCBs in ballast sediments were estimated to have median or high potential of posing ecological risks, respectively, to marine life if ballast sediments were disposed without specific treatment. POPs in ballast sediments were derived from multiple sources with atmospheric deposition being an important origin. Ship activities including diesel exhaust and illegal oil sewage discharge were considerable contributors of certain individual POPs to ballast sediments. Our study is important because it represents the first report on levels, health risk assessment and source apportionments of POPs in ballast sediments and is a first step in the implementation of specific ballast sediment management measures.

Pollution characteristics and ecological risk of heavy metals in ballast tank sediment.

This study was conducted to illustrate the contents and potential ecological risk of heavy metals in ballast tank sediment. Ballast sediment samples were collected from six ships during their stay in shipyard, and the heavy metals were determined by inductive coupled plasma emission spectrometer. Results showed that high concentrations of heavy metals were detected in all six sediment samples following the order: Zn > Cu > Pb > Cr > As > Cd > Hg. The geoaccumulation index explained the average pollution degree of heavy metals decreased as the following: Zn > Pb > Cu > As > Cr > Hg, and the environmental risk indices suggested that concentration found of Zn, Pb, and Cu might be highly toxic to aquatic organisms. Principal component and correlation analysis indicated the metal pollution in ballast tank sediment was affected by complex and different contamination mechanisms, and the corrosion of ballast tank played an important role in this process. In conclusion, this study is very useful for comprehensive consideration and efficient management of ballast tank sediment in order to protect the marine environment.

Speciation and transformation of heavy metals during vermicomposting of animal manure.

This work was conducted to evaluate the effects of vermicomposting on the speciation and mobility of heavy metals (Zn, Pb, Cr, and Cu) in cattle dung (CD) and pig manure (PM) using tessier sequential extraction method. Results showed that the pH, total organic carbon and C/N ratio were reduced, while the electric conductivity and humic acid increased after 90days vermicomposting. Moreover, the addition of earthworm could accelerate organic stabilization in vermicomposting. The total heavy metals in final vermicompost from CD and PM were higher than the initial values and the control without worms. Sequential extraction indicated that vermicomposting decreased the migration and availability of heavy metals, and the earthworm could reduce the mobile fraction, while increase the stable fraction of heavy metals. Furthermore, these results indicated that vermicomposting played a positive role in stabilizing heavy metals in the treatment of animal manure.

Pyrosequencing reveals bacterial community differences in composting and vermicomposting on the stabilization of mixed sewage sludge and cattle dung.

This study aimed to compare the microbial community structures and compositions in composting and vermicomposting processes. We applied 454 high-throughput pyrosequencing to analyze the 16S rRNA gene of bacteria obtained from bio-stabilization of sewage sludge and cattle dung. Results demonstrated that vermicomposting process presented higher operational taxonomic units and bacterial diversity than the composting. Analysis using weighted UniFrac indicated that composting exhibited higher effects on shaping microbial community structure than the vermicomposting. The succession of dominant bacteria was also detected during composting. Firmicutes was the dominant bacteria in the thermophilic phase of composting and shifted to Actinomycetes in the maturing stage. By contrast, Proteobacteria accounted for the highest proportions in the whole process of the vermicomposting. Furthermore, vermicomposting contained more uncultured and unidentified bacteria at the taxonomy level of genus than the composting. In summary, the bacterial community during composting significantly differed from that during vermicomposting. These two techniques played different roles in changing the diversity and composition of microbial communities.