Bacterial Community Resistance Research Articles

Gut bacterial consortium enriched in a biofloc system protects shrimp against Vibrio parahaemolyticus infection

BackgroundShrimp cultured in a biofloc system (BFS) have a lower disease incidence than those farmed in a water exchange system (WES). Although a number of studies have reported that the gut bacterial community induced by BFS is highly associated with shrimp disease resistance, the causal relationship remains unknown. Here, the promotive roles of gut bacterial community induced by BFS in pathogenic Vibrio infection resistance and its potential micro-ecological and physiological mechanisms were investigated by gut bacterial consortium transplantation and synthetic community (SynCom) construction.ResultsThe BFS induced a more stable and resistant gut bacterial community, and significantly enriched some beneficial bacterial taxa, such as Paracoccus, Ruegeria, Microbacterium, Demequina, and Tenacibaculum. Transplantation of a gut bacterial consortium from BFS shrimp (EnrichBFS) greatly enhanced the stability of the bacterial community and resistance against pathogenic V. parahaemolyticus infection in WES shrimp, while transplantation of a gut bacterial consortium from WES shrimp significantly disrupted the bacterial community and increased pathogen susceptibility in both WES and BFS shrimp. The addition of EnrichBFS in shrimp postlarvae also improved the pathogen resistance through increasing the relative abundances of beneficial bacterial taxa and stability of bacterial community. The corresponding strains of five beneficial bacterial taxa enriched in BFS shrimp were isolated to construct a SynComBFS. The addition of SynComBFS could not only suppress disease development, but also improve shrimp growth, boost the digestive and immune activities, and restore health in diseased shrimp. Furthermore, the strains of SynComBFS well colonized shrimp gut to maintain a high stability of bacterial community.ConclusionsOur study reveals an important role for native microbiota in protecting shrimp from bacterial pathogens and provides a micro-ecological regulation strategy towards the development of probiotics to ameliorate aquatic animal diseases.1NpdyYjfCEzGqNeY6CWHQHVideo

Marine resistome of a temperate zone: Distribution, diversity, and driving factors across the trophic gradient

Marine and ocean environments are the most widespread habitats in the world but are still the least studied from the aspect of antibiotic resistance. The indigenous and tetracycline (TET)- and sulfamethoxazole (SXT)-resistant planktonic bacterial communities were simultaneously investigated for the first time along a trophic gradient of a temperate zone, regarding their taxonomic and functional structures as well as biotic and abiotic factors affecting their dynamics as vehicles of antibiotic resistance genes (ARGs), thus impacting the ARGs distribution at seasonal and spatial scales.A total of 80 microbiomes, recovered seasonally from bottom layer and surface waters along a 68-km transect from wastewater-impacted estuary to coastal and pristine open sea in the central Adriatic (Mediterranean Sea), were analysed using 16S rRNA amplicon sequencing, PICRUSt2 bioinformatics and extensive biostatistics. Eighty-one bacterial phyla were identified, with majority (n = 49) in summer when communities were found to be more species enriched across the gradient. Microbial diversity was more site-specific and pronounced in surface microbiomes in winter. Nevertheless, both richness and community diversity decreased with distance from the coast. Although the microbiomes from human-influenced sites significantly differed from those in oligotrophic offshore area, Proteobacteria were still the most abundant phylum during both seasons at the surface and seabed along the gradient, and the major contributors to the marine resistome regarding native and TET- and SXT-resistant microbial communities. Resistome structure was more diverse in winter, whereas peptide, vancomycin and multidrug resistance modules predominated regardless of season, trophic status, or antibiotic. However, multidrug, beta-lactam resistance modules as well as macrolide, phenicol, aminoglycoside, and particularly imipenem resistance genes were much more frequent in winter, suggesting that the diversity of indigenous resistomes is highly dependent on seasonal variations of the water column, driven by thermohaline stratification and nutrients. Moreover, several pathogenic genera stood out as important carriers of multiple resistance traits in TET- and SXT-related resistomes in both seasons, particularly Acinetobacter, Vibrio, Bacillus and Pseudomonas, beside which Proteus, Serratia and Bacteroides prevailed in native resistomes.This study evidenced seasonal and spatial variations of the marine microbiome and resistome and their driving forces along the trophic gradient, providing a comprehensive insight into the diversity and distribution of antibiotic resistance in the marine ecosystem of a temperate zone.

Antibiotic resistance in wastewater: Indian scenario

The surge of Antibiotic Resistant Bacteria (ARB) in the environment is poised to be the next health threat. World Health Organisation's (WHO's) Global Antimicrobial Surveillance System (GLASS) report indicates that developing countries may be at a greater risk. Among various factors, the major driver here could be untreated wastewater and poor sanitation. Bacteria are extremely adaptable to their surroundings and develop Antimicrobial Resistance (AMR) when exposed to antibiotics and other pollutants that cause microbial stress. Thus, untreated domestic wastewater drains could easily become hotspots for the occurrence of ARBs. This study reports surveillance of sewage-carrying drains across four urban cities in India and demonstrated the presence of ARBs in the bacterial community against 7 classes of antibiotics, namely, β-Lactams, Chloramphenicol, Glycopeptides, Macrolides, Tetracycline, Third Generation Cephalosporin, and Quinolones. Untreated domestic wastewater flowing in target drains was collected twice a month, for a period of six months and the microbial community was subjected to Antibiotic Susceptibility Testing (AST) by plate assays. The zone of inhibition was recorded and interpreted as per the interpretive chart of The Clinical & Laboratory Standards Institute (CLSI) & The European Committee on Antimicrobial Susceptibility Testing (EUCAST). The total number of samples showing resistance against antibiotics was used to define an Antibiotic Resistance Index (ARI), calculated for all 20 sampling sites (drains). Results demonstrated that the highest ARI was observed in Delhi and Mumbai, ranging from 0.81 to 0.92 in Delhi and 0.49–0.56 in Mumbai. This surveillance study reveals the antibiotic resistance pattern of the representative bacterial community in the drains and goes beyond few targeted bacterial species. The alarming presence of antibiotic resistant bacterial community highlights the concern of ARBs being the next looming health threat. This report aims to demonstrates the importance of considering sewage surveillance on routine basis by state authorities.

Complex therapy of recurrent urinary infections

Recurrent urinary tract infections (RUTI) occur in 10-15 % of women of reproductive age and in 20 % of menopausal women and have an extremely negative impact on quality of life. Urinary tract infections (UTIs) account for about 40 % of all hospital-acquired infections and 50% of bacterial infections resulting in long-term hospitalisation. In Russia, 26-36 million cases of acute cystitis are reported each year. Because of its high prevalence, UTI is an economic problem. Clinically, UTI is more often manifested as acute urethritis or cystitis. Antibiotic therapy is a fundamental treatment for RUTI, but has a number of negative consequences. Antimicrobial resistance is an enormous challenge and a serious threat to global health, prompting the search for alternative strategies. Natural antimicrobial peptides have attracted the attention of researchers and clinicians for their broad therapeutic potential against bacteria, viruses and resistant intracellular bacterial communities (biofilms). This review details the etiology, classification, diagnosis and therapy of RUTI. Global data revealing the mechanisms of action of antimicrobial peptides is presented. Commercially available peptide-based antimicrobials and their efficacy based on clinical trials are discussed. The development of new strategies to combat bacterial infections, in particular the use of drugs based on antimicrobial peptides and their derivatives, has been shown to be of great practical importance in terms of combating multidrug-resistant pathogens and increasing the effectiveness of treatment of recurrent infections.

The Addition of Biochar and Hyper-Thermal Inoculum Can Regulate the Fate of Heavy Metals Resistant Bacterial Communities during the Livestock Manure Composting

In the present investigation the effects of biochar and hyper-thermal inoculum on the heavy-metal-resistant bacteria (HMRB) during livestock manure composting were studied. An experiment was performed on composting livestock manure and wheat straw amended with biochar and hyper-thermal inoculum. Physicochemical properties, enzyme activity, heavy metals (HMs), and bacterial activities were monitored, and a comprehensive assessment was analyzed during the composting process. The results showed that the dominant phyla of Proteobacteria, Bacteroidota, Actinobacteriota, and Chloroflexi were enriched, but this was not the case with Firmicutes. The abundance of Galbibacter, Thermobifida, Sphaerobacter, and Actinomadura was significantly different in CT15 and BHCT15. In addition, this study showed that the selected factors are less correlated with HMRB compared with the CT group. Therefore, this study could provide new insights into the effect of biochar and hyper-thermal inoculum amendments on the fate of HMRB under HMs and high temperature stress during livestock manure composting.

High stability and metabolic capacity of bacterial community promote the rapid reduction of easily decomposing carbon in soil

Irreversible climate change alters the decomposition and sequestration of soil carbon (C). However, the stability of C components in soils with different initial organic matter contents and its relationship with the response of major decomposers to climate warming are still unclear. In this study, we translocated Mollisols with a gradient of organic matter (OM) contents (2%–9%) from in situ cold region to five warmer climatic regions to simulate climate change. Soil C in C-rich soils (OM >5%) was more vulnerable to translocation warming than that in C-poor soils (OM ≤ 5%), with a major loss of functional groups like O-alkyl, O-aryl C and carboxyl C. Variations of microbial β diversity with latitude, temperature and precipitation indicated that C-rich soils contained more resistant bacterial communities and more sensitive fungal communities than C-poor soils, which led to strong C metabolism and high utilization ability of the community in C-rich soils in response to translocation warming. Our results suggest that the higher sensitivity of soils with high organic matter content to climate change is related to the stability and metabolic capacity of major bacterial decomposers, which is important for predicting soil-climate feedback.

Contribution of enrofloxacin and Cu2+ to the antibiotic resistance of bacterial community in a river biofilm

Pollutants discharged from wastewater are the main cause of the spread of antibiotic resistance in river biofilms. There is controversy regarding the primary contribution of environmental selectors such as antibiotics and heavy metals to the development of antibiotic resistance in bacterial communities. Here, this study compared the effect of environmental safety concentration Cu2+ and enrofloxacin (ENR) on the evolution of antibiotic resistance by examining phenotypic characteristics and genotypic profiles of bacterial communities in a river biofilm, and then distinguished the major determinants from a comprehensive perspective. The pollution induced community tolerance in ENR-treated group was significantly higher than that in Cu2+-treated group (at concentration levels of 100 and 1000 μg/L). Metagenomic sequencing results showed that ENR significantly increased the number and total abundance of antibiotic resistance genes (ARGs), but there was no significant change in the Cu2+- treated group. Compared with Cu2+, ENR was the major selective agent in driving the change of taxonomic composition because the taxonomic composition in ENR was the most different from the original biofilm. Comparing and analyzing the prokaryotic composition, the phylum of Proteobacteria was enriched in both ENR and Cu2+ treated groups. Among them, Acidovorax and Bosea showed resistance to both pollutants. Linking taxonomic composition to ARGs revealed that the main potential hosts of fluoroquinolone resistance genes were Comamonas, Sphingopyxis, Bradyrhizobium, Afipia, Rhodopseudomonas, Luteimonas and Hoeflea. The co-occurrence of ARGs and metal resistance genes (MRGs) showed that the multidrug efflux pump was the key mechanism connecting MRGs and ARGs. Network analysis also revealed that the reason of Cu2+ selected for fluoroquinolones resistant bacterial communities was the coexistence of multidrug efflux gene and MRGs. Our research emphasizes the importance of antibiotics in promoting the development of antibiotic resistant bacterial communities from the perspective of changes in community structure and resistome in river biofilms.

Patterns of heavy metal resistant bacterial community succession influenced by biochar amendment during poultry manure composting

The purpose of this study was to investigate the heavy metal resistant bacteria (HMRB) community succession and bacterial activity in poultry manure (PM) composting. Five different concentration of chicken manure biochar (CMB) at 0%, 2%, 4%, 6%, and 10% on a dry weight basis was applied with initial feedstock (poultry manure + wheat straw) and indicated with T1, T2, T3, T4, and T5. In addition, high-throughput sequencing, principal coordinate analysis, and correlation analysis were used to analyze the evolution of HMRB communities during composting. The study indicated that crucial phyla were Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes. The bacterial diversity in the CMB amendment treatment was higher than in the control treatment, and T4 treatment has the highest among all CMB applied treatments. Moreover, results from CCA indicated that T4 and T5 treatments quickly enters the high-temperature period which is maintained for 5 days, and is significantly positively correlated with Proteobacteria, and Actinobacteria. These findings offer insight into potential strategies to understand the succession of HMRBs during PM reuse. Overall, the above results show the addition of 6% biochar (T4) was potentially beneficial to enrich the abundance of bacterial community to improve composting environment quality and composting efficiency. In addition, effective to immobilized the heavy metals and HMRB in the end product.

Distribution of heavy metal resistant bacterial community succession in cow manure biochar amended sheep manure compost

The aim of this investigation was to study the effects of cow manure biochar (CMB) on the distribution of heavy metal resistant bacterial (HMRB) community succession during sheep manure (SM) composting. The experiments were conducted with six different ratio of CMB (0%(T1), 2.5%(T2), 5%(T3),7.5%(T4),10%(T5) and 12%(T6)onadryweightbasis) and 0% is used as control. The results showed that the most dominant phylum were Proteobacteria (40.89%-5.65%) and Firmicutes (0.16%-93.18%), and 7.5% CMB mixed with sheep manure for best results. Thus, significant correlation was noticed among the analyzed physicochemical factors, gaseous emission and bacterial phylum in used 7.5–10% CMB applied for SM composting. Overall, the application of biochar increased the diversity of the bacterial community and promoted the degradation of organic matter. In addition, 7.5–10% CMB applied treatments showed greater immobilization of HMRB community succession during SM composting.

Can biochar regulate the fate of heavy metals (Cu and Zn) resistant bacteria community during the poultry manure composting?

In this study, the influence of coconut shell biochar addition (CSB) on heavy metals (Cu and Zn) resistance bacterial fate and there correlation with physicochemical parameters were evaluated during poultry manure composting. High-throughput sequencing was carried out on five treatments, namely T1−T5, where T2 to T5 were supplemented with 2.5%, 5%, 7.5% and 10% CSB, while T1 was used as control for the comparison. The results of HMRB indicated that the relative abundance of major potential bacterial host altered were Firmicutes (52.88–14.32%), Actinobacteria (35.20–4.99%), Bacteroidetes (0.05–15.07%) and Proteobacteria (0.01–20.28%) with elevated biochar concentration (0%−10%). Beta and alpha diversity as well as network analysis illustrated composting micro-environmental ecology with exogenous additive biochar to remarkably affect the dominant resistant bacterial community distribution by adjusting the interacting between driving environmental parameters with potential host bacterial in composting. Ultimately, the amendment of 7.5% CSB into poultry manure composting was able to significantly reduce the HMRB abundance, improve the composting efficiency and end product quality.

The influence of genetically modified glyphosate-tolerant maize CC-2 on rhizosphere bacterial communities revealed by MiSeq sequencing

Genetically modified (GM) crops have brought huge economic benefits to mankind, however, at the same time, their safety issues are drawing growing attention. This investigation was conducted to assess whether the long-term cultivation of GM glyphosate resistant maize CC-2 effects bacterial communities in the rhizosphere soil. A 2-year follow-up trial was conducted, and soils were sampled at various plant developmental stages. The bacterial community structure of the rhizosphere soil was analysed by the high-throughput sequencing and compared with the near-isogenic non-GM maize Zheng 58. We showed here that long-term cultivation of CC-2 has no significant effect on the structure and diversity of bacterial communities, while different growth stages had significant effect. These results provided a reliable theoretical basis for the future cultivation and increased commercialisation of CC-2.

The relationship between culturable doxycycline-resistant bacterial communities and antibiotic resistance gene hosts in pig farm wastewater treatment plants

Pig farm wastewater treatment plants (WWTPs) are an important repository for resistant bacterial communities (RBCs) and antibiotic resistance genes (ARGs). However, the relationship between RBCs and ARG hosts has not been well characterized. In this study, water samples from influent and effluent from five pig farm WWTPs were collected. Gradient concentrations of doxycycline (DOX) were used to screen the culturable RBCs. The abundance of 21 subtypes of ARGs and the bacterial community were investigated. This study detected a large number of culturable DOX-RBCs and ARGs in the influent and effluent of pig farm WWTPs. The abundances of ARGs and RBCs in all effluent samples was significantly lower than that in the influent samples (P < 0.05), which indicated that the WWTPs can effectively remove most ARGs and RBCs in pig farm wastewater. The main potential culturable RBCs in pig farm wastewater were the dominant bacteria Proteobacteria, Actinobacteria, Pseudomonas, and Rheinheimera. However, most of the ARGs were mainly present in Bacteroidetes, Actinobacteria, Corynebacteriaceae, Macellibacteroides, Acinetobacter, and Enterobacteriaceae, which are considered potential ARG hosts. The results presented here showed that there were obvious differences between the species of culturable DOX-RBCs and ARG hosts in the pig farm WWTPs, which may be due to various environmental factors. This highlights the urgent need for further research on the relationship between RBCs and ARG hosts.

Isolation of Arsenic Resistant and Arsenopyrite Oxidizing Acidithiobacillus Species from pH Neutral Colombian Mine Effluents

Inactive mines provide a great source of bacterial diversity for studying acidophilic communities and their biotechnological applications, but prospecting of these anthropogenic environments in Colombia has been limited. Conventional microbiological methods were used to isolate acidophilic bacterial strains from effluents emanating from the Colombian gold mine ‘El Zancudo’ (Titiribí, Antioquia). Despite the drainage waters having circumneutral pH, all of the isolated strains were phylogenetically related to the extreme acidophile Acidithiobacillus genus. However, based upon 16S rRNA gene sequences the mesophilic sulfur-oxidizing indigenous strains could not be assigned to a species. Pure cultures were selected by screening in medium with soluble inorganic arsenic (III) and their mineral-oxidative activity was evaluated at 30 °C in Erlenmeyer flasks with arsenopyrite ore under rotary shaking conditions. The indigenous strains were able to catalyze arsenopyrite oxidation in a mixed culture with a pulp density of 10%, maintaining their growth in the presence of >80 mM leached arsenic. This research provides information regarding the isolation of arsenic resistant bacterial communities from neutral effluents from El Zancudo mine and the possibility of the isolated strains to be useful in the biooxidation pretreatment of refractory gold-bearing arsenopyrite ores and concentrates.

Roles of different humin and heavy-metal resistant bacteria from composting on heavy metal removal

This study aims to assess the roles of different humin and heavy-metal resistant bacterial community from composting on heavy metal removal. The results showed that the concentration of Cu2+, Zn2+, Ni2+, Pb2+, Cr3+ and Cd2+ decreased with adding the compost-derived humin, but the removal rates were relatively low (<30% on average). The heavy metal resistant bacteria from composting have better metal binding capacities than humin, and the combined addition of humin and bacteria could further stimulate the biosorption of heavy metals with 60–80% removal of metals and improve the diversity and biomass of bacterial community. There was obviously increased synergy between the humin from maturity phase and bacteria for metal biosorption (“1 + 1 > 2”). Structural equation modeling showed that microbial biomass and humin humification are the key factors for the biosorption of heavy metals. Combining humin from maturity phase with heavy-metal resistant bacteria was suggested to control heavy metal pollution in composts.

Selectively enrichment of antibiotics and ARGs by microplastics in river, estuary and marine waters

The partition of antibiotics and antibiotic resistant genes (ARGs) between the microplastics (MPs) and the surrounding water with various salinity are still unclear. In this study, we hypothesized that adsorption of antibiotics on MPs might cause a significant change of the structure of microbial communities, diversity and abundance of ARGs on MPs and this might be further affected by change of salinity. In this study, we investigated adsorption of four common antibiotics (sulfamerazine, tetracycline, chloramphenicol and tylosin) to polyethylene (PE) MPs in river, estuary and marine waters, and the differences of antibiotic resistant genes (ARGs) and bacterial communities on MPs and in the three waters. The results showed that MPs can enrich antibiotics, ARGs and microbes from the surrounding water. Elevated salinity could reduce adsorption of antibiotics to MPs and the abundance of ARGs. For example, MPs can concentrate more antibiotics and ARGs in the fresh river water than in the estuary and the marine waters. In addition, ARGs and bacterial communities on MPs at various salinity were significantly different under the pressure of four antibiotics. On MPs, sul1, sulA/folP-01, tetA, tetC, tetX and ermE increased significantly but a few new ARGs such as sulA/folP-01 and tetA appeared. The structure of the bacterial communities on MPs was different from the surrounding water since some bacteria species found on MPs were barely detected in the surrounding water while some genera on MPs vanished after exposure to antibiotics. As the antibiotics adsorbed and the ARGs on MPs decreased with the water salinity, the structure of the communities on MPs thus varied with salinity change. These findings are important to understand the effects of MPs on the transport, fate and ecological risk of antibiotics and ARGs in different aquatic environments.

Effects of zinc orthophosphate on the antibiotic resistant bacterial community of a source water used for drinking water treatment

The presence of zinc, and not orthophosphate, was the selective pressure for antibiotic resistance.

Antimicrobial assessment of phage therapy using a porcine model of biofilm infection

Antibiotic resistant bacterial communities persist in many types of wounds, chronic wounds in particular, in the form of biofilms. Biofilm formation is a major cause of severe infections and the main reason for a negative treatment outcome and slow healing progression. Chronic wounds are a silent epidemic essentially affecting people with co-morbid conditions such as diabetes and obesity and elderly persons particularly those with movement limitations. The development of complementary and alternative effective strategies to antibiotics for the treatment of chronic wounds is highly desired. Phage therapy constitutes a very promising approach in the control of topical microbial populations. In this work newly isolated phages were tested for their efficacy to control bacterial species that predominate in chronic wounds. Phage effectiveness was studied on 24-h old biofilms formed in polystyrene microplates and in porcine skin explants using two treatment approaches: individual phage and a cocktail of phages against four main pathogens commonly isolated from chronic wounds. The two models produced variations in the surface colonization ability, assessed by viable bacterial counts and microscopy visualization after using peptide nucleic acid (PNA) or locked nucleic acid probes (LNA) and 2′-O-methyl (2′-OMe) in fluorescence in situ hybridization (FISH), and in the phage-host interactions. Phages alone and combined caused greater reductions in the number of viable cells when biofilms had been formed on porcine skins and with greater variations detected at 4 h and 24 h of sampling. These results suggest that porcine skin models should be preferentially used to assess the use of phages and phage cocktails intended for topical use in order to understand the fate, throughout treatment time, of the population when dealing with biofilm-related infections.

Biosurfactant coated silver and iron oxide nanoparticles with enhanced anti-biofilm and anti-adhesive properties.

Pseudomonas aeruginosa and Staphylococcus aureus are among the hazardous biofilm forming bacteria ubiquitous in industrial/clinical wastes. Serious efforts are required to develop effective strategies to control surface-growing antibiotic resistant pathogenic bacterial communities which they are emerging as a global health issue. Blocking hazardous biofilms would be a useful aspect of biosurfactant coated nanoparticles (NPs). In this regard, we report a facile method for the synthesis of rhamnolipid (RL) coated silver (Ag) and iron oxide (Fe3O4) NPs and propose the mechanism of their synergistic antibacterial and anti-adhesive properties against biofilms formed by P. aeruginosa and S. aureus. These NPs demonstrated excellent anti-biofilm activity not only during the biofilms formation but also on the pre-formed biofilms. Mechanistically, RL coated silver (35 nm) and Fe3O4 NPs (48 nm) generate reactive oxygen species, which contribute to the antimicrobial activity. The presence of RLs shell on the nanoparticles significantly reduces the cell adhesion by modifying the surface hydrophobicity and hence enhancing the anti-biofilm property of NPs against both mentioned strains. These findings suggest that RL coated Ag and Fe3O4 NPs may be used as potent alternate to reduce the infection severity by inhibiting the biofilm formation and, therefore, they possess potential biomedical applications for antibacterial coatings and wound dressings.

Enhanced organics and Cu2+ removal in electroplating wastewater by bioaugmentation

With the improvement of electroplating process and products requirement, refractory organics, heavy metals or even heavy metal nanoparticles (NP) exist simultaneously in electroplating wastewater inevitably, makes electroplating wastewater treatment effluent difficult to meet the discharge standard. In order to improve the organics removal under the exposure of CuO NP, strains (designated as L1–L5) that have both organics degradation and Cu2+ tolerance capacities were isolated and employed in the electroplating wastewater bioaugmentation treatment using a hydrolytic/anoxic/oxic-membrane bioreactor. The Cu2+ adsorption process followed pseudo-second order kinetics and the isotherms fit well to Langmuir isotherm model. L2, L3 and L4 showed higher Cu2+ adsorption capacity than that of L1 and L5. Under the optimal condition, the maximum Cu2+ adsorption capacity of L2, L3 and L4 was 34.15, 45.68 and 26.72 mg g−1, respectively. Their average COD removal efficiency achieved 65.7 ± 10.9%, 61.5 ± 6.7% and 71.6 ± 6.0%, respectively. The three isolates were used to construct consortia with the inoculum concentration of 400 mg L−1. One-time and repeated inoculations were evaluated to find the applicable strategy. Repeated inoculation resulted in a better COD and Cu removal performance (76.2 ± 2.6% and 98.5 ± 0.3%, respectively) than those of one-time inoculation (69.0 ± 2.0% and 98.0 ± 0.3%, respectively). The most functionally stable, balanced and resistant bacterial community was formed in the one-time inoculation system while for fungal community it was formed in the repeated inoculation system.

Modeling the Emergence of Antibiotic Resistance in the Environment: an Analytical Solution for the Minimum Selection Concentration.

Environmental antibiotic risk management requires an understanding of how subinhibitory antibiotic concentrations contribute to the spread of resistance. We develop a simple model of competition between sensitive and resistant bacterial strains to predict the minimum selection concentration (MSC), the lowest level of antibiotic at which resistant bacteria are selected. We present an analytical solution for the MSC based on the routinely measured MIC, the selection coefficient (sc) that expresses fitness differences between strains, the intrinsic net growth rate, and the shape of the bacterial growth dose-response curve with antibiotic or metal exposure (the Hill coefficient [κ]). We calibrated the model by optimizing the Hill coefficient to fit previously reported experimental growth rate difference data. The model fit varied among nine compound-taxon combinations examined but predicted the experimentally observed MSC/MIC ratio well (R2 ≥ 0.95). The shape of the antibiotic response curve varied among compounds (0.7 ≤ κ ≤ 10.5), with the steepest curve being found for the aminoglycosides streptomycin and kanamycin. The model was sensitive to this antibiotic response curve shape and to the sc, indicating the importance of fitness differences between strains for determining the MSC. The MSC can be >1 order of magnitude lower than the MIC, typically by the factor scκ This study provides an initial quantitative depiction and a framework for a research agenda to examine the growing evidence of selection for resistant bacterial communities at low environmental antibiotic concentrations.