Heavy Metal Resistance Research Articles

Environmentally Relevant Antibiotic Concentrations Exert Stronger Selection Pressure on River Biofilm Resistomes than AMR-Reservoir Effluents.

Freshwater environments are primary receiving systems of wastewater and effluents, which carry low concentrations of antibiotics and antimicrobial-resistant (AMR) bacteria and genes. Aquatic microbial communities are thus exposed to environmentally relevant concentrations of antibiotics (ERCA) that presumably influence the acquisition and spread of environmental AMR. Here, we analyzed ERCA exposure with and without the additional presence of municipal wastewater treatment plant effluent (W) and swine manure run-off (M) on aquatic biofilm resistomes. Microscopic analyses revealed decreased taxonomic diversity and biofilm structural integrity, while metagenomic analysis revealed an increased abundance of resistance, virulence, and mobile element-related genes at the highest ERCA exposure levels, with less notable impacts observed when solely exposed to W or M effluents. Microbial function predictions indicated increased gene abundance associated with energy and cell membrane metabolism and heavy metal resistance under ERCA conditions. In silico predictions of increased resistance mechanisms did not correlate with observed phenotypic resistance patterns when whole communities were exposed to antimicrobial susceptibility testing. This reveals important insight into the complexity of whole-community coordination of physical and genetic responses to selective pressures. Lastly, the environmental AMR risk assessment of metagenomic data revealed a higher risk score for biofilms grown at sub-MIC antibiotic conditions.

Selective Isolation and Identification of Microorganisms with Dual Capabilities: Leather Biodegradation and Heavy Metal Resistance for Industrial Applications.

Tanning, crucial for leather production, relies heavily on chromium yet poses risks due to chromium's oxidative conversion, leading to significant wastewater and solid waste generation. Physico-chemical methods are typically used for heavy metal removal, but they have drawbacks, prompting interest in eco-friendly biological remediation techniques like biosorption, bioaccumulation, and biotransformation. The EU Directive (2018/850) mandates alternatives to landfilling or incineration for industrial textile waste management, highlighting the importance of environmentally conscious practices for leather products' end-of-life management, with composting being the most researched and viable option. This study aimed to isolate microorganisms from tannery wastewater and identify those responsible for different types of tanned leather biodegradation. Bacterial shifts during leather biodegradation were observed using a leather biodegradation assay (ISO 20136) with tannery and municipal wastewater as the inoculum. Over 10,000 bacterial species were identified in all analysed samples, with 7 bacterial strains isolated from tannery wastewaters. Identification of bacterial genera like Acinetobacter, Brevundimonas, and Mycolicibacterium provides insights into potential microbial candidates for enhancing leather biodegradability, wastewater treatment, and heavy metal bioremediation in industrial applications.

Phenotypic characterization for bioremediation suitability of isolates from Southern Tunisian tannery effluent

Effluents from the leather tanning industry contain diverse pollutants, including hazardous heavy metals, posing threats to public health and the surrounding environment. Indigenous bacterial isolates can represent an eco-friendly approach for tannery wastewater treatment; however, phenotypic characterization is necessary to determine whether these strains are suitable for bioremediation. In the present study, we analyzed seven new Enterococcus faecium strains and two new Bacillus subtillis strains isolated from effluents from the Southern Tunisian Tannery (ESTT). We evaluated phenotypic features beneficial for bioremediation, including biofilm formation, hydrophobicity, and exoenzyme activities. Additionally, we examined characteristics naturally occurring in environmental bacteria but less desirable in strains selected for bioremediation, such as antibiotic resistances and pathogenicity indicators. The observed phenotypes were then compared with whole-genome analysis. We observed biofilm production in two slime-producing bacteria, B. licheniformis RLT6, and E. faecium RLT8. Hydrophobicity of E. faecium strains RLT1, RLT5, RLT8, and RLT9, as well as B. licheniformis RLT6 correlated positively with increasing ESTT concentration. Exoenzyme activities were detected in E. faecium strains RLT2, RLT4, and RLT7, as well as B. licheniformis RLT6. As anticipated, all strains exhibited common resistances to antibiotics and hemolysis, which are widespread in nature and do not hinder their application for bioremediation. Importantly, none of the strains exhibited the pathogenic hypermucoviscosity phenotype. To the best of our knowledge, this is the first report consolidating all these phenotypic characteristics concurrently, providing a complete overview of strains suitability for bioremediation. ImportanceThe study evaluates the bioremediation potential of seven Enterococcus faecium strains and two Bacillus subtillis strains isolated from the effluents from the Southern Tunisian tannery (ESTT), which pose threats to public health and environmental integrity. The analysis primarily examines the phenotypic traits crucial to bioremediation, including biofilm formation, hydrophobicity, and exoenzyme activities, as well as characteristics naturally occurring in environmental bacteria related to heavy metal resistance, such as antibiotic resistances. Several strains were found to have high bioremediation potential and exhibit only antibiotic resistances commonly found in nature, ensuring their application for bioremediation remains uncompromised. The results of the exhaustive phenotypic analysis are contrasted with the whole genome sequences of the nine strains, underscoring the appropriateness of these bacterial strains for eco-friendly interventions in tannery wastewater treatment.

A comprehensive comparative genomic analysis revealed that plant growth promoting traits are ubiquitous in strains of Stenotrophomonas.

Stenotrophomonas strains, which are often described as plant growth promoting (PGP) bacteria, are ubiquitous in many environments. A total of 213 genomes of strains of Stenotrophomonas were analyzed using comparative genomics to better understand the ecological roles of these bacteria in the environment. The pan-genome of the 213 strains of Stenotrophomonas consists of 27,186 gene families, including 710 core gene families, 11,039 unique genes and 15,437 accessory genes. Nearly all strains of Stenotrophomonas harbor the genes for GH3-family cellulose degradation and GH2- and GH31-family hemicellulose hydrolase, as well as intact glycolysis and tricarboxylic acid cycle pathways. These abilities suggest that the strains of this genus can easily obtain carbon and energy from the environment. The Stenotrophomonas strains can respond to oxidative stress by synthesizing catalase, superoxide dismutase, methionine sulfoxide reductase, and disulfide isomerase, as well as managing their osmotic balance by accumulating potassium and synthesizing compatible solutes, such as betaine, trehalose, glutamate, and proline. Each Stenotrophomonas strain also contains many genes for resistance to antibiotics and heavy metals. These genes that mediate stress tolerance increase the ability of Stenotrophomonas strains to survive in extreme environments. In addition, many functional genes related to attachment and plant colonization, growth promotion and biocontrol were identified. In detail, the genes associated with flagellar assembly, motility, chemotaxis and biofilm formation enable the strains of Stenotrophomonas to effectively colonize host plants. The presence of genes for phosphate-solubilization and siderophore production and the polyamine, indole-3-acetic acid, and cytokinin biosynthetic pathways confer the ability to promote plant growth. These strains can produce antimicrobial compounds, chitinases, lipases and proteases. Each Stenotrophomonas genome contained 1-9 prophages and 17-60 genomic islands, and the genes related to antibiotic and heavy metal resistance and the biosynthesis of polyamines, indole-3-acetic acid, and cytokinin may be acquired by horizontal gene transfer. This study demonstrates that strains of Stenotrophomonas are highly adaptable for different environments and have strong potential for use as plant growth-promoting bacteria.

Physiological and transcriptomic analyses reveal the cadmium tolerance mechanism of Miscanthus lutarioriparia.

Miscanthus lutarioriparia is a promising energy crop that is used for abandoned mine soil phytoremediation because of its high biomass yield and strong tolerance to heavy metals. However, the biological mechanism of heavy metal resistance is limited, especially for applications in the soil restoration of mining areas. Here, through the investigation of soil cadmium(Cd) in different mining areas and soil potted under Cd stress, the adsorption capacity of Miscanthus lutarioriparia was analyzed. The physiological and transcriptional effects of Cd stress on M. lutarioriparia leaves and roots under hydroponic conditions were analyzed. The results showed that M. lutarioriparia could reduce the Cd content in mining soil by 29.82%. Moreover, different Cd varieties have different Cd adsorption capacities in soils with higher Cd concentration. The highest cadmium concentrations in the aboveground and belowground parts of the plants were 185.65 mg/kg and 186.8 mg/kg, respectively. The total chlorophyll content, superoxide dismutase and catalase activities all showed a trend of increasing first and then decreasing. In total, 24,372 differentially expressed genes were obtained, including 7735 unique to leaves, 7725 unique to roots, and 8912 unique to leaves and roots, which showed differences in gene expression between leaves and roots. These genes were predominantly involved in plant hormone signal transduction, glutathione metabolism, flavonoid biosynthesis, ABC transporters, photosynthesis and the metal ion transport pathway. In addition, the number of upregulated genes was greater than the number of downregulated genes at different stress intervals, which indicated that M. lutarioriparia adapted to Cd stress mainly through positive regulation. These results lay a solid foundation for breeding excellent Cd resistant M. lutarioriparia and other plants. The results also have an important theoretical significance for further understanding the detoxification mechanism of Cd stress and the remediation of heavy metal pollution in mining soil.

Biodegradation and antimicrobial capability-induced heavy metal resistance of the marine-derived actinomycetes Nocardia harenae JJB5 and Amycolatopsis marina JJB11.

Currently, heavy metal-resistant (HMR) marine actinomycetes have attracted much attention worldwide due to their unique capabilities. In this study, 27 marine-derived actinomycetes were isolated from coastal beaches in the Arabian Gulf of Al-Jubail in Saudi Arabia and screened for resistance to 100 mg/L of the heavy metals Cd2+, Cr6+, Cu2+, Fe2+, Pb2+, and Ni2+ using different assay techniques. Six isolates were selected as HMRs, of which two isolates, JJB5 and JJB11, exhibited the highest maximum tolerance concentrations (200- > 300 mg/L). Both isolates were the highest among six-HMR screened for their biodegradation potential of plastics low-density polyethylene, polystyrene, and polyvinyl chloride, recording the highest weight loss (15 ± 1.22 - 65 ± 1.2%) in their thin films. They also showed the highest biodegradability of the pesticides acetamiprid, chlordane, hexachlorocyclohexane, indoxacarb and lindane, indicating promising removal capacities (95.70-100%) for acetamiprid and indoxacarb using HPLC analysis. Additionally, the cell-free filtrate (CFF) of both isolates displayed the highest antimicrobial activity among the six-HMR screened against a variety of microbial test strains, recording the highest inhibition zone diameters (13.76 ± 0.66 - 26.0 ± 1.13 mm). GC‒MS analyses of the ethyl acetate extract of their CFFs revealed the presence of diverse chemical compounds with a multitude of remarkable biological activities. Based on their spore morphology and wall-chemotype, they were assigned to the nocardioform-actinomycetes. Furthermore, their phenotypic characteristics, together with 16S rRNA gene sequencing (OR121525-OR121526), revealed them as Nocardia harenae JJB5 and Amycolatopsis marina JJB11. Our results suggest that marine HMR actinomycetes are promising candidates for various biotechnological applications.

Actinomycetes as a Possible Source of Bioremediation of Heavy Metal Cadmium from Contaminated Soil

Annually, a significant volume of chemicals, encompassing fertilizers and pesticides, is administered to agricultural soils. Using pesticides and fertilizers, agricultural practices contribute to heavy metal Cadmium (Cd), Copper (Cu), Lead (Pb) and Zinc (Zn) pollution. Heavy metals and pesticides are high at the peak of ecological contaminants, presence of this has introduced grave risks to the health of the population and agronomics. Among heavy metals, cadmium (Cd) is recognized for its high mobility in various environmental settings. Cd has a deleterious effect on plant phenotypic, cytotoxicity (e.g., lowering chlorophyll concentration and limiting photosynthetic effectiveness), and metabolic activities (e.g., chlorosis and necrosis). Microbial bioremediation by using microorganisms is one of the secure, pure, cost operative and eco-friendly technology for decontaminating polluting sites as compared to physical and chemical techniques. Among microbes, Actinobacteria hold a paramount position, serving as key players in numerous biological processes, utilize toxins as carbon source and turn into high concentrations of pesticides, chemical complexes and heavy metals into commercially viable antibiotics, enzymes, proteins, and plant growth promoting hormones. This study is an effort to explore the potent cadmium resistance actinomycetes to reduce cadmium levels to enhance degradation. For this purpose, 53 actinomycetes strains were tested for heavy metal resistance and tolerance to Cadmium against different concentrations. After secondary screening Four potent isolate have the potential to grow at 1000 mg/L concentration of Cadmium in the medium. When they are able to grow on heavy metal containing media it could be beneficial for reduction and elimination of toxic metals from contaminated environment. When it comes to achieving a suitable level of metal tolerance, this potent powerful actinomycetes strain Streptomyces pactum have been identified to be promising.

Copper Resistance Mechanism and Copper Response Genes in Corynebacterium crenatum.

Heavy metal resistance mechanisms and heavy metal response genes are crucial for microbial utilization in heavy metal remediation. Here, Corynebacterium crenatum was proven to possess good tolerance in resistance to copper. Then, the transcriptomic responses to copper stress were investigated, and the vital pathways and genes involved in copper resistance of C. crenatum were determined. Based on transcriptome analysis results, a total of nine significantly upregulated DEGs related to metal ion transport were selected for further study. Among them, GY20_RS0100790 and GY20_RS0110535 belong to transcription factors, and GY20_RS0110270, GY20_RS0100790, and GY20_RS0110545 belong to copper-binding peptides. The two transcription factors were studied for the function of regulatory gene expression. The three copper-binding peptides were displayed on the C. crenatum surface for a copper adsorption test. Furthermore, the nine related metal ion transport genes were deleted to investigate the effect on growth in copper stress. This investigation provided the basis for utilizing C. crenatum in copper bioremediation.

Omics technology draws a comprehensive heavy metal resistance strategy in bacteria.

The rapid industrial revolution significantly increased heavy metal pollution, becoming a major global environmental concern. This pollution is considered as one of the most harmful and toxic threats to all environmental components (air, soil, water, animals, and plants until reaching to human). Therefore, scientists try to find a promising and eco-friendly technique to solve this problem i.e., bacterial bioremediation. Various heavy metal resistance mechanisms were reported. Omics technologies can significantly improve our understanding of heavy metal resistant bacteria and their communities. They are a potent tool for investigating the adaptation processes of microbes in severe conditions. These omics methods provide unique benefits for investigating metabolic alterations, microbial diversity, and mechanisms of resistance of individual strains or communities to harsh conditions. Starting with genome sequencing which provides us with complete and comprehensive insight into the resistance mechanism of heavy metal resistant bacteria. Moreover, genome sequencing facilitates the opportunities to identify specific metal resistance genes, operons, and regulatory elements in the genomes of individual bacteria, understand the genetic mechanisms and variations responsible for heavy metal resistance within and between bacterial species in addition to the transcriptome, proteome that obtain the real expressed genes. Moreover, at the community level, metagenome, meta transcriptome and meta proteome participate in understanding the microbial interactive network potentially novel metabolic pathways, enzymes and gene species can all be found using these methods. This review presents the state of the art and anticipated developments in the use of omics technologies in the investigation of microbes used for heavy metal bioremediation.

Habitat- and lifestyle-dependent structural and functional characteristics of viruses in mangrove wetlands of different functional zonings

Mangrove wetlands, as one of the natural ecosystems with the most ecological services, have garnered widespread attention about their microbial driven biogeochemical cycling. Urbanization have led to different spatial patterns of environmental conditions and microbial communities in mangroves. However, viruses, as the pivotal drivers of biogeochemical cycling in mangroves, remain inadequately explored in terms of how their ecological potential and complex interactions with host respond to functional zonings. To address this knowledge gap, we conducted a comprehensive investigation on the structural and functional properties of temperate and lytic viruses in mangrove wetlands from different functional zonings by jointly using high-throughput sequencing, prokaryotic and viral metagenomics. Multiple environmental factors were found to significantly influence the taxonomic and functional composition, as well as lysogen-lysis decision-making of mangrove viruses. Furthermore, enriched auxiliary metabolic genes (AMGs) involved in methane, nitrogen and sulfur metabolism, and heavy metal resistance were unveiled in mangrove viruses, whose community composition was closely related to lifestyle and host. The virus-host pairs with different lifestyles were also discovered to react to environmental changes in different ways, which provided an empirical evidence for how virus and bacteria dynamics were specific to viral lifestyles in nature. This study expands our comprehension of the intricate interactions among virus, prokaryotic host and the environment in mangrove wetlands from multiple perspectives, including viral lifestyles, virus-host interactions, and habitat dependence. Importantly, it provides a new ecological perspective on how mangrove viruses are adapted to the stress posed by urbanization.

Isolating Indigenous Metal Resistant Gram-Positive Bacteria: Assessing Their Selenite Reduction Potential

The objective of this study is to isolate gram-positive bacilli from soil and to study the effects of physiological parameters on their heavy metal resistance. High levels of metals in the soil can have major toxic effects on soil microorganisms and may cause a great change in the species diversity and population sizes of soil microbes. In current study soil samples were collected from the agriculture drainage water and soil in Lahore. After the selection of sixty gram-positive strains, their biochemical and metabolic profile was structured and all were auxin producing. The isolates were resistant to multiple heavy metals including Ni. Pb, Cr, Li and Hg. Their selenite reduction was estimated at varyingpH, Temperature and concentrations of selenite and all were resistant upto 3000 ?g/ml, However, their efficacy of reducing selenite was majorly affected by set physiological parameters.

Genotypic analyses of IncHI2 plasmids from enteric bacteria

Incompatibility (Inc) HI2 plasmids are large (typically > 200 kb), transmissible plasmids that encode antimicrobial resistance (AMR), heavy metal resistance (HMR) and disinfectants/biocide resistance (DBR). To better understand the distribution and diversity of resistance-encoding genes among IncHI2 plasmids, computational approaches were used to evaluate resistance and transfer-associated genes among the plasmids. Complete IncHI2 plasmid (N = 667) sequences were extracted from GenBank and analyzed using AMRFinderPlus, IntegronFinder and Plasmid Transfer Factor database. The most common IncHI2-carrying genera included Enterobacter (N = 209), Escherichia (N = 208), and Salmonella (N = 204). Resistance genes distribution was diverse, with plasmids from Escherichia and Salmonella showing general similarity in comparison to Enterobacter and other taxa, which grouped together. Plasmids from Enterobacter and other taxa had a higher prevalence of multiple mercury resistance genes and arsenic resistance gene, arsC, compared to Escherichia and Salmonella. For sulfonamide resistance, sul1 was more common among Enterobacter and other taxa, compared to sul2 and sul3 for Escherichia and Salmonella. Similar gene diversity trends were also observed for tetracyclines, quinolones, β-lactams, and colistin. Over 99% of plasmids carried at least 25 IncHI2-associated conjugal transfer genes. These findings highlight the diversity and dissemination potential for resistance across different enteric bacteria and value of computational-based approaches for the resistance-gene assessment.

Genomic and environmental controls on Castellaniella biogeography in an anthropogenically disturbed subsurface

Castellaniella species have been isolated from a variety of mixed-waste environments including the nitrate and multiple metal-contaminated subsurface at the Oak Ridge Reservation (ORR). Previous studies examining microbial community composition and nitrate removal at ORR during biostimulation efforts reported increased abundances of members of the Castellaniella genus concurrent with increased denitrification rates. Thus, we asked how genomic and abiotic factors control the Castellaniella biogeography at the site to understand how these factors may influence nitrate transformation in an anthropogenically impacted setting. We report the isolation and characterization of several Castellaniella strains from the ORR subsurface. Five of these isolates match at 100% identity (at the 16S rRNA gene V4 region) to two Castellaniella amplicon sequence variants (ASVs), ASV1 and ASV2, that have persisted in the ORR subsurface for at least 2 decades. However, ASV2 has consistently higher relative abundance in samples taken from the site and was also the dominant blooming denitrifier population during a prior biostimulation effort. We found that the ASV2 representative strain has greater resistance to mixed metal stress than the ASV1 representative strains. We attribute this resistance, in part, to the large number of unique heavy metal resistance genes identified on a genomic island in the ASV2 representative genome. Additionally, we suggest that the relatively lower fitness of ASV1 may be connected to the loss of the nitrous oxide reductase (nos) operon (and associated nitrous oxide reductase activity) due to the insertion at this genomic locus of a mobile genetic element carrying copper resistance genes. This study demonstrates the value of integrating genomic, environmental, and phenotypic data to characterize the biogeography of key microorganisms in contaminated sites.

Transcriptomic and physiological analyses of Symphytum officinale L. in response to multiple heavy metal stress

Soil heavy metal contamination has become a global environmental issue, which threaten soil quality, food security and human health. Symphytum officinale L. have exhibited high tolerance and restoration capacity to heavy metals (HMs) stress. However, little is known about the mechanisms of HMs in S. officinale. In this study, transcriptomic and physiological changes of S. officinale response to different HMs (Pb, Cd and Zn) were analyzed and investigated the key genes and pathways involved in HMs uptake patterns. The results showed that phenotypic effects are not significant, and antioxidant enzyme activities were all upregulated. Transcriptome analysis indicated that 1247 differential genes were up-regulated, and 1963 differential genes were down-regulated under Cd stress, while 3752 differential genes were up-regulated, and 7197 differential genes were down-regulated under Pb stress; and 527 differential genes were up-regulated; and 722 differential genes were down-regulated under Zn stress. Based on their expression, we preliminarily speculate that different HMs resistance of S. officinale may be regulated by the differential expression of key genes. These results provide a theoretical basis for determining the exact expression of genes in plants under different heavy metal stress, the processes involved molecular pathways, and how they can be efficiently utilized to improve plant tolerance to toxic metals and improve phytoremediation efficiency.

Effect of aqueous phase from hydrothermal carbonization of sewage sludge on heavy metals and heavy metal resistance genes during chicken manure composting

Livestock manure is often contaminated with heavy metals (HMs) and HM resistance genes (HMRGs), which pollute the environment. In this study, we aimed to investigate the effects of the aqueous phase (AP) produced by hydrothermal carbonization (HTC) of sewage sludge (SS) alone and the AP produced by co-HTC of rice husk (RH) and SS (RH-SS) on humification, HM bioavailability, and HMRGs during chicken manure composting. RH-SS and SS increased the humic acid content of the compost products by 18.3 % and 9.7 %, respectively, and significantly increased the humification index (P < 0.05) compared to the CK (addition of tap water). The passivation of HMs (Zn, Cu, As, Pb, and Cr) increased by 12.17–23.36 % and 9.74–15.95 % for RH-SS and SS, respectively, compared with that for CK. RH-SS and SS reduced the HMRG abundance in composted products by 22.29 % and 15.07 %, respectively. The partial least squares path modeling results showed that SS and RH-SS promoted compost humification while simultaneously altering the bacterial community and reducing the bioavailability of metals and host abundance of HMRGs, which has a direct inhibitory effect on the production and distribution of HMRGs. These findings support a new strategy to reduce the environmental risk of HMs and HMRGs in livestock manure utilization.

Strategic implementation of upflow microbubble airlift photocatalytic process to control heavy metal resistant-MDR bacteria and the associated genes in wastewater

The emergence of multidrug resistance (MDR) and heavy metal resistance (HMR) in bacterial pathogens and genes through mobile genetic elements (MGEs) from wastewater effluents needs to be eradicated using green technology. HMR is critical to controlling antimicrobial resistance but has been neglected in most photocatalytic disinfection (PCD) studies. Advanced photocatalysis in antimicrobial studies requires a strategic approach to enhance the disinfection efficiency. This study applied an optimized upflow airlift photocatalytic reactor (UAPR) with microbubble air and blue light to remove the antimicrobial resistance (HMR–MDR bacteria (HMR–MDRB), HMR genes (HMRGs), antibiotic resistant genes (ARGs) and MGEs) and inhibit pathogenicity in a mixed culture of high-strength HMR–MDRB isolated from wastewater. Stable dissolved oxygen (DO) supplementation and blue light significantly affected the HMRGs and MGE reduction, resulting in enhanced disinfection activity (30 %) and reaction time (60 min) required for the complete inactivation of HMR-MDRB mixed culture. The process effectively reduced the blaNDM-1 (3.37 log), pbrT (2.18 log), intl1 (2.16 log), and tn916/1545 (4.07 log), with a PCD rate constant (kg) of 0.0515 min−1, 0.0359 min−1, 0.0292 min−1, and 0.0662 min−1, respectively. Process mechanism studies showed that the stable supplementation of DO was a critical factor in controlling the type of reactive oxygen species (ROS), resulting in type II ROS, 1O2, as the critical free radical contributing to the reduction of ARGs and integron, whereas h+ was the ROS for pbrT and transposon.

Effects of removing in-feed antibiotics and zinc oxide on the taxonomy and functionality of the microbiota in post weaning pigs

BackgroundPost weaning diarrhoea (PWD) causes piglet morbidity and mortality at weaning and is a major driver for antimicrobial use worldwide. New regulations in the EU limit the use of in-feed antibiotics (Ab) and therapeutic zinc oxide (ZnO) to prevent PWD. New approaches to control PWD are needed, and understanding the role of the microbiota in this context is key. In this study, shotgun metagenome sequencing was used to describe the taxonomic and functional evolution of the faecal microbiota of the piglet during the first two weeks post weaning within three experimental groups, Ab, ZnO and no medication, on commercial farms using antimicrobials regularly in the post weaning period.ResultsDiversity was affected by day post weaning (dpw), treatment used and diarrhoea but not by the farm. Microbiota composition evolved towards the dominance of groups of species such as Prevotella spp. at day 14dpw. ZnO inhibited E. coli overgrowth, promoted higher abundance of the family Bacteroidaceae and decreased Megasphaera spp. Animals treated with Ab exhibited inconsistent taxonomic changes across time points, with an overall increase of Limosilactobacillus reuteri and Megasphaera elsdenii. Samples from non-medicated pigs showed virulence-related functions at 7dpw, and specific ETEC-related virulence factors were detected in all samples presenting diarrhoea. Differential microbiota functions of pigs treated with ZnO were related to sulphur and DNA metabolism, as well as mechanisms of antimicrobial and heavy metal resistance, whereas Ab treated animals exhibited functions related to antimicrobial resistance and virulence.ConclusionAb and particularly ZnO maintained a stable microbiota composition and functionality during the two weeks post weaning, by limiting E. coli overgrowth, and ultimately preventing microbiota dysbiosis. Future approaches to support piglet health should be able to reproduce this stable gut microbiota transition during the post weaning period, in order to maintain optimal gut physiological and productive conditions.

Isolation and Characterization of Heavy Metal-Tolerant Bacterial Isolates from Industrial Effluents in Uttar Pradesh, India

Industrial activities over the past century have significantly increased human exposure to pollutants such as heavy metals. Industrial emissions of heavy metals, which are carcinogenic, mutagenic and toxic, contaminate natural water supplies and the agricultural environment. Due to the high concentration of heavy metals in industrial effluents, the bacteria present there naturally develop resistance to heavy metals. The aim of this study is to isolate and characterize bacteria resistant to heavy metal, lead (Pb), copper (Cu), chromium (Cr), cadmium (Cd), and nickel (Ni) from sites contaminated by industrial effluents in Uttar Pradesh, India. A total of 58 bacterial isolates were isolated from 9 samples according to their various morphological parameters and 32 isolates were found positive for all heavy metals, which revealed that samples contained metal-resistant bacterial diversity. Two isolates were identified up to species level based on their physiological, biochemical and molecular characterization as Comamonas testosteroni (S4C1) and Bacillus cereus (S5C3). Both isolates are highly resistant to lead (Pb), copper (Cu), chromium (Cr), cadmium (Cd), and nickel (Ni) and they show different MICs against the above heavy metals at different levels. A growth experiment showed that the presence of heavy metals concentration had no discernible impact on the growth rate among the isolates. Gel analysis showed interesting patterns of protein expression were observed in the presence of various heavy metals. MALDI-TOF analysis found that specific proteins (S layer protein, F0F1ATP synthase subunit b, Flagellin, 50S ribosomal protein L4, Molecular chaperone) were overexpressed in the presence of heavy metals. As a result, identifying the heavy metal resistance bacteria and their proteins study could be useful as a preliminary investigation for the development of prospective bioremediation agents of potentially hazardous waste treatment technology.

Crucifer Lesion-Associated Xanthomonas Strains Show Multi-Resistance to Heavy Metals and Antibiotics.

Copper resistance in phytopathogens is a major challenge to crop production globally and is known to be driven by excessive use of copper-based pesticides. However, recent studies have shown co-selection of multiple heavy metal and antibiotic resistance genes in bacteria exposed to heavy metal and xenobiotics, which may impact the epidemiology of plant, animal, and human diseases. In this study, multi-resistance to heavy metals and antibiotics were evaluated in local Xanthomonas campestris pv. campestris (Xcc) and co-isolated Xanthomonas melonis (Xmel) strains from infected crucifer plants in Trinidad. Resistance to cobalt, cadmium, zinc, copper, and arsenic (V) was observed in both Xanthomonas species up to 25 mM. Heavy metal resistance (HMR) genes were found on a small plasmid-derived locus with ~ 90% similarity to a Stenotrophomonas spp. chromosomal locus and a X. perforans pLH3.1 plasmid. The co-occurrence of mobile elements in these regions implies their organization on a composite transposon-like structure. HMR genes in Xcc strains showed the lowest similarity to references, and the cus and ars operons appear to be unique among Xanthomonads. Overall, the similarity of HMR genes to Stenotrophomonas sp. chromosomal genomes suggest their origin in this genus or a related organism and subsequent spread through lateral gene transfer events. Further resistome characterization revealed the presence of small multidrug resistance (SMR), multidrug resistance (MDR) efflux pumps, and bla (Xcc) genes for broad biocide resistance in both species. Concurrently, resistance to antibiotics (streptomycin, kanamycin, tetracycline, chloramphenicol, and ampicillin) up to 1000 µg/mL was confirmed.

Long-term and combined heavy-metal contamination forms a unique microbiome and resistome: A case study in a Yellow River tributary sediments

Microorganisms have developed mechanisms to adapt to environmental stress, but how microbial communities adapt to long-term and combined heavy-metal contamination under natural environmental conditions remains unclear. Specifically, this study analyzed the characteristics of heavy metal composition, microbial community, and heavy metal resistance genes (MRGs) in sediments along Mang River, a tributary of the Yellow River, which has been heavily polluted by industrial production for more than 40 years. The results showed that the concentrations of Cr, Zn, Pb, Cu and As in most sediments were higher than the ambient background values. Bringing the heavy metals speciation and concentration into the risk evaluation method, two-thirds of the sediment samples were at or above the moderate risk level, and the ecological risk of combined heavy metals in the sediments decreased along the river stream. The high ecological risk of heavy metals affected the microbial community structure, metabolic pathways and MRG distribution. The formation of a HM-resistant microbiome possibly occurred through the spread of insertion sequences (ISs) carrying multiple MRGs, the types of ISs carrying MRGs outnumber those of plasmids, and the quantity of MRGs on ISs is also higher than that on plasmids. These findings could improve our understanding of the adaptation mechanism of microbial communities to long-term combined heavy metal contamination.