Abundance Of Tetracycline Resistance Genes Research Articles

Polyvinyl chloride microplastics in the aquatic environment enrich potential pathogenic bacteria and spread antibiotic resistance genes in the fish gut

Microplastics and antibiotics coexist in aquatic environments, especially in freshwater aquaculture areas. However, as the second largest production of polyvinyl chloride (PVC) in the world, the effects of co-exposure to microplastics particles and antibiotics on changes in antibiotic resistance gene (ARG) profiles and the microbial community structure of aquatic organism gut microorganisms are poorly understood. Therefore, in this study, carp (Cyprinus carpio) were exposed to single or combined PVC microplastic contamination and oxytetracycline (OTC) or sulfamethazine (SMZ) for 8 weeks. PVC microplastics can enrich potential pathogenic bacteria, such as Enterobacter and Acinetobacter, among intestinal microorganisms. The presence of PVC microplastics enhanced the selective enrichment and dissemination risk of ARGs. PVC microplastics combined with OTC (OPVC) treatment significantly increased the abundance of tetracycline resistance genes (1.40-fold) compared with that in the OTC exposure treatment, revealing an obvious co-selection effect. However, compared with those in the control group, the total abundance of ARGs and MGEs in the OPVC treatment groups were significantly lower, which was correlated with the reduced abundances of the potential host Enterobacter. Overall, our results emphasized the diffusion and spread of ARGs are more influenced by PVC microplastics than by antibiotics, which may lead to antibiotic resistance in aquaculture.

Multi-omics analysis of gut microbiota and metabolites reveals contrasting profiles in domestic pigs and wild boars across urban environments.

The gut microbiota plays a crucial role in host health and metabolism. This study explores the differences in gut microbiota and metabolites between domestic pigs (DP) and wild boars (WB) in urban environments. We analyzed gut microbial composition, metabolic profiles, virome composition, antibiotic resistance genes (ARGs), and human pathogenic bacteria (HPB) in both DP and WB. Our results revealed that DP exhibited a higher Firmicutes/Bacteroidetes ratio and were enriched in bacterial genera associated with domestication and modern feeding practices. Metabolomic analysis showed distinct profiles, with WB significantly enriched in the Pantothenate and CoA biosynthesis pathway, highlighting dietary and environmental influences on host metabolism. Additionally, DP had a distinct gut virome composition, particularly enriched in lytic phages of the Chaseviridae family. ARG analysis indicated a higher abundance of tetracycline resistance genes in DP, likely due to antibiotic use in pig farms. Furthermore, variations in HPB composition underscored potential health risks associated with contact with pig feces. These findings provide valuable insights into the microbial ecology of domestic pigs and wild boars, emphasizing the importance of these comparisons in identifying zoonotic pathogen transmission pathways and managing antibiotic resistance. Continued research in this area is essential for developing effective strategies to mitigate public health risks and promote sustainable livestock management practices.

Transmission of antimicrobial resistance in the gut microbiome of gregarious cockroaches: the importance of interaction between antibiotic exposed and non-exposed populations.

Antimicrobial resistance (AMR) is a major global health concern, further complicated by its spread via the microbiome bacterial members. While mathematical models discuss AMR transmission through the symbiotic microbiome, experimental studies are scarce. Herein, we used a gregarious cockroach, Pycnoscelus surinamensis, as an in vivo animal model for AMR transmission investigations. We explored whether the effect of antimicrobial treatment is detectable with metagenomic sequencing, and whether AMR genes can be spread and established in unchallenged (not treated with antibiotics) individuals following contact with treated donors, and under various frequencies of interaction. Gut and soil substrate microbiomes were investigated by metagenomic sequencing for bacterial community composition and resistome profiling. We found that tetracycline treatment altered the treated gut microbiome by decreasing bacterial diversity and increasing the abundance of tetracycline resistance genes. Untreated cockroaches that interacted with treated donors also had elevated tetracycline resistance. The levels of resistance differed depending on the magnitude and frequency of donor transfer. Additionally, treated donors showed signs of microbiome recovery due to their interaction with the untreated ones. Similar patterns were also recorded in the soil substrate microbiomes. Our results shed light on how interacting microbiomes facilitate AMR gene transmission to previously unchallenged hosts, a dynamic influenced by the interaction frequencies, using an in vivo model to validate theoretical AMR transmission models.IMPORTANCEAntimicrobial resistance is a rising threat to human and animal health. The spread of resistance through the transmission of the symbiotic gut microbiome is of concern and has been explored in theoretical modeling studies. In this study, we employ gregarious insect populations to examine the emergence and transmission of antimicrobial resistance in vivo and validate modeling hypotheses. We find that antimicrobial treatment increases the levels of resistance in treated populations. Most importantly, we show that resistance increased in untreated populations after interacting with the treated ones. The level of resistance transmission was affected by the magnitude and frequency of population mixing. Our results highlight the importance of microbial transmission in the spread of antimicrobial resistance.

Encapsulated peracetic acid as a valid broad-spectrum antimicrobial alternative, leading to beneficial microbiota compositional changes and enhanced performance in broiler chickens

BackgroundAntimicrobial alternatives are urgently needed, including for poultry production systems. In this study, we tested the potential broad-range antimicrobial alternative peracetic acid, delivered in feed via the hydrolysis of encapsulated precursors through a 28-day study using 375 Ross 308 broiler chickens. We tested two peracetic acid concentrations, 30 and 80 mg/kg on birds housed on re-used litter, and we evaluated the impact of both levels on gut microbial communities, bacterial concentration, antimicrobial resistance genes relative abundance and growth performance when compared to control birds housed on either clean or re-used litter.ResultsBody weight gain and feed conversion ratio improved in peracetic acid fed birds. At d 28, birds given 30 mg/kg of peracetic acid had a decreased Firmicutes and an increased Proteobacteria abundance in the jejunum, accompanied by an increase in Bacillus, Flavonifractor and Rombustia in the caeca, and a decreased abundance of tetracycline resistance genes. Chicken given 80 mg/kg of peracetic acid had greater caecal abundance of macrolides lincosamides and streptogramins resistance genes. Growth performance on clean litter was reduced compared to re-used litter, which concurred with increased caecal abundance of Blautia, decreased caecal abundance of Escherichia/Shigella, Anaerostipes and Jeotgalicoccus, and greater gene abundance of vancomycin, tetracycline, and macrolides resistance genes.ConclusionsPeracetic acid could be used as a safe broad-spectrum antimicrobial alternative in broilers. Encapsulated precursors were able to reduce the bacterial concentration in the jejunum whilst promoting the proliferation of probiotic genera in the caeca, especially at the low peracetic acid concentrations tested, and improve growth performance. Moreover, our findings offer further insights on potential benefits of rearing birds on re-used litter, suggesting that the latter could be associated with better performance and reduced antimicrobial resistance risk compared to clean litter rearing.

Sulfonamide and tetracycline resistance genes in nanjing lakes: effect of water quality and heavy metals

Due to high population density, anthropogenic activities and waste disposal have negatively affected artificial lakes in residential areas. These artificial lakes lack the capacity for auto-depuration to remove pollutants and contaminants; thus, they have the potential to act as reservoirs for antibiotic resistance genes (ARGs). In this study, we examined three urban artificial freshwater lakes in Nanjing to determine the abundance of sulfonamide and tetracycline resistance genes. Concerning water quality, the three lakes were found to be highly eutrophic, owing to their high levels of Total Nitrogen (TN), Phosphorous (TP), and Chlorophyll a (Chla). The average abundance of sulfonamide resistance genes detected in the three urban lakes was 42.446 log10 gene copies/100 ml, which was lower than the average abundance of tetracycline resistance genes (68.207 log10 gene copies/100 ml). Analysis by ANOVA revealed that all ARGs, except sul3, showed significant differences, probably due to varied anthropogenic influences in lakes. Pearson correlation and principal component analyses were performed to explore the correlation between ARGs, water quality markers, and heavy metals to understand the co-selection and drivers of ARGs propagation. tetM showed no correlation with any water quality markers, whereas Chla showed a positive correlation with all ARGs except tetM. tetM was the only gene observed to be unaffected by TN, TP, and Chla. The tet genes also showed strong associations with each other except tetM, especially tetA, tetQ, and tetG. The co-selection results between heavy metals and ARGs were insignificant (p > 0.05), with tetM being the most sensitive to the effects of heavy metals and As having the strongest effect on sul3 and tet genes. The results from this study provide basic but archival information on the effect of eutrophication and heavy metals such as Arsenic, showing the potential influence on the dissemination of certain sulfonamide and tetracycline ARGs in freshwater environments.

Metagenomic insights into the influence of mobile genetic elements on ARGs along typical wastewater treatment system on pig farms in China

The wastewater treatment processes (WTP) on pig farms are heavily contaminated by antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) play an important role in shaping ARG profiles. Here we first employed metagenomic sequencing to follow the diversities and shifts of ARG associated mobile genetic elements (AAMGEs) including insertion sequences (ISs) and plasmids along the WTP for three pig farms in southeast China. The IS average relative abundance rose from the initial pig feces source to the wastewater storage lagoon (WSL) but decreased in the influent and rose in the effluent of the anaerobic digestor (AD). In contrast, plasmids were eliminated rapidly along this process. These results indicated that the AD reduced plasmid copies while IS abundance increased. We found a great diversity ISs, including IS91, ISNCY, IS630 and IS701, were large contributors to the transfer of multi-drug resistance. In addition, the tetracycline resistance genes co-occurred with a greater diversity of ISs than other ARG classes and this likely contributed to the high abundance of tetracycline resistance genes we found. The transfer of ARGs mediated by MGEs along the WTP of pig farms was a key contributor for the ARGs persistence in the environment of pig farms. Collectively, our findings demonstrated different fates for ISs and plasmids along the WTP for pig farms and suggested that AAMGE monitoring served as an important role in controlling ARGs in pig waste.

Faecal microbiota transplantation alleviates early-life antibiotic-induced gut microbiota dysbiosis and mucosa injuries in a neonatal piglet model

Faecal microbiota transplantation (FMT) is a promising approach to modulate the gut microbiota. Gut microbiota dysbiosis caused by antibiotic administration is a universal problem. This study aimed to evaluate the effect of FMT on the dysbiosis of gut microbiota and metabolic profiles and injury of the intestinal barrier induced by antibiotics and used a neonatal piglet model. Neonatal piglets were administered ampicillin for 3 days, and antibiotic-induced dysbiosis was evaluated by the occurrence of diarrhoea and alteration of gut microbiota. Then, FMT was conducted for 3 days to rebuild the gut microbiota. High-throughput sequencing and a mass spectrometry platform were used for integrated microbiome-metabolome analysis. The results showed that antibiotics led to a decline in the diversity of gut microbiota. Furthermore, there was an increase in the relative abundance of potential pathogenic bacteria, such as Oscillibacter, Pseudomonas and Eubacterium, and an increase in the relative abundance of tetracycline resistance genes (tet genes). FMT restored the diversity and promoted the relative abundance of beneficial bacteria, such as Parabacteroides, Dorea and Parasutterella, while decreasing the relative abundance of tet genes. Untargeted metabolomics analysis found that alpha linolenic acid and linoleic acid metabolism were the key metabolic pathways utilized in the FMT group, and targeted metabolomics analysis further verified the variation in the associated metabolites arachidonic acid and conjugated linoleic acid. FMT also significantly enhanced the relative expression of tight junction (ZO-1, claudin-1 and occludin) and adherens junction (β-catenin, E-cadherin) proteins and anti-inflammatory cytokines (IL-10, TGF-β1) and reduced the production of proinflammatory cytokines (IL-6, IL-1β, TNF-α and IFN-γ) in the colon. FMT not only modulated the gut microbiota composition and microbial metabolism but also reduced the relative abundance of tet genes, improving the intestinal barrier function and inflammatory responses in antibiotic-treated piglets.

Changes of antibiotic resistance genes and bacterial communities in the advanced biological wastewater treatment system under low selective pressure of tetracycline

Effluents of conventional wastewater treatment systems contain antibiotic residues at concentrations below the minimal inhibitory concentrations (MIC), which nevertheless could still select for antibiotic-resistant bacteria. This work focuses on evaluating the changes of antibiotic resistance genes (ARGs) and bacterial communities in a planted advanced biological wastewater treatment system (ABWWTS) under long-term exposure to sub-MIC tetracycline. In the ABWWTS, the removal rates of tetracycline ranged from 97.9% to 99.9%, and a 17.2% decrease in the average removal rates of NH4+-N was observed after the addition of tetracycline. Although the background of ABWWTS contributed to the ARGs in effluents, the concentration of 283 targeted ARGs (ΣARGs) was 83.5% lower in effluents than in influents after sub-MIC tetracycline exposure, and the concentrations of ΣARGs in the ABWWTS were, on average, 30.0% lower than those in an unplanted biological wastewater treatment system (UBWWTS) after a performance of 130 days. The relative abundance of tetracycline resistance genes increased within ABWWTS and UBWWTS under tetracycline exposure. After tetracycline exposure, bacterial diversity in ABWWTS and UBWWTS increased on average by 36.2% and 42.7%, respectively, and the abundances of Nitrosomonas and Nitrospira in the aerobic zone were more than 10-times higher in the ABWWTS than in the UBWWTS. Sub-MIC tetracycline concentrations were linearly correlated with the relative abundance of tetracycline resistance genes in Escherichia coli (E. coli). Long-term exposure to tetracycline at the same concentration increased abundances of the same ARGs (i.e., tetR-02 and tetM-01) in E. coli and the microflora of the ABWWTS, revealing that sub-MIC tetracycline could increase the abundance of ARGs in the ABWWTS by facilitating the vertical transfer of tetracycline resistance genes. These findings demonstrated that planted ABWWTS played a positive role in removing ARGs under low antibiotic selective pressure, which was in accompany with increasing levels of corresponding ARGs within the system.

Effects of different oxygen conditions on pollutants removal and the abundances of tetracycline resistance genes in activated sludge systems

The individual and combined effects of tetracycline (TC) and divalent copper (Cu2+) on the performance of activated sludge systems and the abundances of tetracycline resistance genes (TRGs) in activated sludge, under both aerobic and anaerobic conditions, were studied. Activated sludge systems received TC (0.2 mg L−1) and Cu2+ (5 mg L−1) separately or jointly under either aerobic or anaerobic conditions. The addition of TC did not affect the performance of activated sludge systems and the addition of Cu2+ and mixed TC/Cu2+ inhibited biological phosphorus removal. The TC removal efficiencies in systems under aerobic and anaerobic conditions were 98.4%–99.7% and 96.8%–99.9%, respectively, and Cu2+ promoted TC removal in activated sludge systems. The TC degradation product was 4-epitetracycline (ETC) in activated sludge systems under both aerobic and anaerobic conditions. The total relative abundances of TRGs (tetA, tetC, tetE, tetM, tetO, tetW, tetX and tetB(P)) in activated sludge showed opposite development trends under the two oxygen conditions and aerobic condition was beneficial to the attenuation of high-risk TRGs. The results of this study might improve evaluation of the combined effects of antibiotics and heavy metals on wastewater biological treatment systems.

Persistence and spread of tetracycline resistance genes and microbial community variations in the soil of animal corrals in a semi-arid planted forest.

At the spring, goat and sheep herds are transferred to planted forests, in a semi-arid region in the northern Negev Desert, Israel, to reduce herbaceous biomass and, fire risk. The herds are held overnight in corrals for about 4 months, enriching the soil with organic matter and nitrogen. This research examined the effect of these enrichments on soil bacterial community structure (BCS) and the abundance of tetracycline resistance genes (TRGs) in active and abandoned corrals (1-10-years-old). Based on 16S rRNA gene sequences, the Proteobacteria and Actinobacteria phyla dominated the soil of all corrals. The Actinobacteria were less abundant in the active and 1-year-old corrals (23-26%) than in the other corrals and the control (33-38%). A principal component analysis showed that, the BCS in the active and the 1-year-old abandoned corrals was significantly different from that in the older corrals and the control. The Firmicutes phylum constituted 28% of the BCS in the active corrals, 12.5% in the 1-year-old corrals and 2% in the older corrals and the control. In contrast, the Acidobacteria phylum was hardly detected in the active and 1-year-old abandoned corrals and constituted 10% of the BCS in the older corrals. Genes conferring resistance to tetracycline were detected in high numbers. The tetG and tetW genes were detected in the active and abandoned corrals (1-10 years). The tetQ gene was detected only in the active and 1-year-old abandoned corrals. None of the genes were detected in the control soil. The three genes were detected outside an active corral, in the downstream section of an ephemeral tributary. The results prove that abandoned and unobserved periodic animal corrals are an environmental reservoir for TRGs.

Toxicity Assessment of Nano-ZnO Exposure on the Human Intestinal Microbiome, Metabolic Functions, and Resistome Using an In Vitro Colon Simulator.

Nano-ZnO, as a commonly used nanomaterial, has been found in drinking water, food, and medicine; therefore, it poses potential health risks via the digestion system. However, little is known about the toxicity of nano-ZnO on the human intestinal microbiome, which plays critical roles in human health. This study comprehensively investigated the impact of nano-ZnO on the human gut microbiome, metabolic functions, and resistome using an in vitro colon simulator. Nano-ZnO induced concentration-dependent decreases in the production of short-chain fatty acids (SCFAs). Metagenomic analysis revealed that nano-ZnO not only led to dose-dependent shifts in the composition and diversity of the gut microbiota but also changed the key functional pathways of the gut microbiome. Although the diversity of the gut microbiota basically recovered after stopping exposure to nano-ZnO, SCFAs still showed a concentration-dependent decrease. Furthermore, although a medium concentration of nano-ZnO (2.5 mg/L) reduced the abundance of many antibiotic resistance genes (ARGs) by inhibiting the growth of related host bacteria, a low concentration of nano-ZnO (0.1 mg/L) greatly enriched the abundance of tetracycline resistance genes. Our findings provide evidence that nano-ZnO can impact the diversity, metabolism, and functional pathways of the human gut microbiome, as well as the gut resistome, highlighting the potential health effects of nanoparticles.

Antibiotic Resistance in Agricultural Soil and Crops Associated to the Application of Cow Manure-Derived Amendments From Conventional and Organic Livestock Farms.

The application of organic amendments to agricultural soil can enhance crop yield, while improving the physicochemical and biological properties of the recipient soils. However, the use of manure-derived amendments as fertilizers entails environmental risks, such as the contamination of soil and crops with antibiotic residues, antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs). In order to delve into these risks, we applied dairy cow manure-derived amendments (slurry, fresh manure, aged manure), obtained from a conventional and an organic farm, to soil. Subsequently, lettuce and wheat plants were grown in the amended soils. After harvest, the abundance of 95 ARGs and MGE-genes from the amended soils and plants were determined by high-throughput qPCR. The structure of soil prokaryotic communities was determined by 16S rRNA amplicon sequencing and qPCR. The absolute abundance of ARGs and MGE-genes differed between treatments (amended vs. unamended), origins of amendment (conventional vs. organic), and types of amendment (slurry vs. fresh manure vs. aged manure). Regarding ARG-absolute abundances in the amendments themselves, higher values were usually found in slurry vs. fresh or aged manure. These abundances were generally higher in soil than in plant samples, and higher in wheat grain than in lettuce plants. Lettuce plants fertilized with conventional amendments showed higher absolute abundances of tetracycline resistance genes, compared to those amended with organic amendments. No single treatment could be identified as the best or worst treatment regarding the risk of antibiotic resistance in soil and plant samples. Within the same treatment, the resistome risk differed between the amendment, the amended soil and, finally, the crop. In other words, according to our data, the resistome risk in manure-amended crops cannot be directly inferred from the analysis of the amendments themselves. We concluded that, depending on the specific question under study, the analysis of the resistome risk should specifically focus on the amendment, the amended soil or the crop.

Metagenomic Quantification of Genes with Internal Standards.

We demonstrate that an assembly-independent and spike-in facilitated metagenomic quantification approach can be used to screen and quantify over 2,000 genes simultaneously, while delivering absolute gene concentrations comparable to those for quantitative PCR (qPCR). DNA extracted from dairy manure slurry, digestate, and compost was spiked with genomic DNA from a marine bacterium and sequenced using the Illumina HiSeq4000. We compared gene copy concentrations, in gene copies per mass of sample, of five antimicrobial resistance genes (ARGs) generated with (i) our quantitative metagenomic approach, (ii) targeted qPCR, and (iii) a hybrid quantification approach involving metagenomics and qPCR-based 16S rRNA gene quantification. Although qPCR achieved lower quantification limits, the metagenomic method avoided biases caused by primer specificity inherent to qPCR-based methods and was able to detect orders of magnitude more genes than is possible with qPCR assays. We used the approach to simultaneously quantify ARGs in the Comprehensive Antimicrobial Resistance Database (CARD). We observed that the total abundance of tetracycline resistance genes was consistent across different stages of manure treatment on three farms, but different samples were dominated by different tetracycline resistance gene families.IMPORTANCE qPCR and metagenomics are central molecular techniques that have offered insights into biological processes for decades, from monitoring spatial and temporal gene dynamics to tracking ARGs or pathogens. Still needed is a tool that can quantify thousands of relevant genes in a sample as gene copies per sample mass or volume. We compare a quantitative metagenomic approach with traditional qPCR approaches in the quantification of ARG targets in dairy manure samples. By leveraging the benefits of nontargeted community genomics, we demonstrate high-throughput absolute gene quantification of all known ARG sequences in environmental samples.

Faecal microbiota and antimicrobial resistance gene profiles of healthy foals.

The human and domestic animal faecal microbiota can carry various antimicrobial resistance genes (ARGs), especially if they have been exposed to antimicrobials. However, little is known about the ARG profile of the faecal microbiota of healthy foals. A high-throughput qPCR array was used to detect ARGs in the faecal microbiota of healthy foals. To characterise the faecal microbiota and ARG profiles in healthy Australian foals aged less than 1month. Observational study. The faecal microbiota and ARG profiles of 37 Thoroughbred foals with no known gastrointestinal disease or antimicrobial treatment were determined using 16S rRNA gene sequencing and a high-throughput ARG qPCR array. Each foal was sampled on one occasion. Firmicutes and Bacteroidetes were dominant in the faecal microbiota. Foals aged 1-2weeks had significantly lower microbiota richness than older foals. Tetracycline resistance genes were the most common ARGs in the majority of foals, regardless of age. ARGs of high clinical concern were rarely detected in the faeces. The presence of ARGs was associated with the presence of class I integron genes. Samples were collected for a case-control study so foals were not sampled longitudinally, and thus the development of the microbiota as individual foals aged could not be proven. The history of antimicrobial treatment of the dams was not collected and may have affected the microbiota of the foals. The ARGs in foal faeces varied concomitantly with age-related microbiota shifts. The high abundance of tetracycline resistance genes was likely due to the dominance of Bacteroides spp.

Tetracycline Induces the Formation of Biofilm of Bacteria from Different Phases of Wastewater Treatment

The study monitored the effect of tetracycline on bacterial biofilm formation and compared biofilm formation by resistant bacterial strains in different phases of the wastewater treatment process in wastewater treatment plant (WWTP). The crystal violet staining method was used to evaluate the biofilm formation. Biofilm-related bacterial properties were characterized by hydrophobicity, autoaggregation and motility tests. The relative abundance of tetracycline resistance genes (tetW, tetM, tetO, tetA and tetB) in wastewaters were subsequently quantified using qPCR. The results show that the isolates from the nitrification tank produce biofilm with up to 10 times greater intensity relative to the isolates from the sedimentation tank. In isolates of Aeromonas sp. from the nitrification tank, increased biofilm production in the occurrence of tetracycline from a concentration of 0.03125 µg/mL was observed. The tetW gene showed the highest relative abundance out of all the tested genes. From the sampling points, its abundance was the highest in the sedimentation tank of the WWTP. Based on these results, it can be assumed that resistant bacteria are able to form a biofilm and sub-inhibitory tetracycline concentrations induce biofilm formation. WWTPs thus represent a reservoir of antibiotic resistance genes and contribute to the spread of resistance in the natural environment.

Hydrothermal pretreatment of oxytetracycline fermentation residue: Removal of oxytetracycline and increasing the potential for anaerobic digestion

The presence of high level of antibiotics in the antibiotic fermentation residue is one of the main reasons that prevent their direct disposal or further use as a resource. In this study, the feasibility of using the hydrothermal pretreatment for removing oxytetracycline (OTC) from its fermentation residue and enhancing anaerobic digestion was evaluated under different temperatures i.e. 110, 130, 150 and 170°C. The results showed that the removal rate of OTC increased as a function of temperature, and hydrothermal treatment at 130°C for 5 min was found sufficient to reduce the concentration of OTC from 3.9 mg/g to less than the detection limit (i.e., 0.25 ng/g). Biochemical methane potential tests showed that the cumulative methane production over 23 d was 73.7, 215.9, 656.8, and 439.0NmL CH4/gVS for the raw residue and the residue treated at 130, 150, and 170°C for 5 min, respectively. At the same time, the abundances of tetracycline resistance genes were reduced by hydrothermal treatments followed by anaerobic digestion. Conclusively, it is suggested that hydrothermal treatment at 150°C for 5 min was found beneficial for OTC fermentation residues ensuring the removal of OTC and further use of the residue for anaerobic digestion.

Construction of a Tetracycline Degrading Bacterial Consortium and Its Application Evaluation in Laboratory-Scale Soil Remediation.

As an environmental pollutant, tetracycline (TC) can persist in the soil for years and damage the ecosystem. So far, many methods have been developed to handle the TC contamination. Microbial remediation, which involves the use of microbes to biodegrade the pollutant, is considered cost-efficient and more suitable for practical application in soil. This study isolated several strains from TC-contaminated soil and constructed a TC-degrading bacterial consortium containing Raoultella sp. XY-1 and Pandoraea sp. XY-2, which exhibited better growth and improved TC degradation efficiency compared with single strain (81.72% TC was biodegraded within 12 days in Lysogeny broth (LB) medium). Subsequently, lab-scale soil remediation was conducted to evaluate its effectiveness in different soils and the environmental effects it brought. Results indicated that the most efficient TC degradation was recorded at 30 °C and in soil sample Y which had relatively low initial TC concentration (around 35 mg/kg): TC concentration decreased by 43.72% within 65 days. Soil properties were affected, for instance, at 30 °C, the pH value of soil sample Y increased to near neutral, and soil moisture content (SMC) of both soils declined. Analysis of bacterial communities at the phylum level showed that Proteobacteria, Bacteroidetes, Acidobacteria, and Chloroflexi were the four dominant phyla, and the relative abundance of Proteobacteria significantly increased in both soils after bioremediation. Further analysis of bacterial communities at the genus level revealed that Raoultella sp. XY-1 successfully proliferated in soil, while Pandoraea sp. XY-2 was undetectable. Moreover, bacteria associated with nitrogen cycling, biodegradation of organic pollutants, soil biochemical reactions, and plant growth were affected, causing the decline in soil bacterial diversity. Variations in the relative abundance of tetracycline resistance genes (TRGs) and mobile gene elements (MGEs) were investigated, the results obtained indicated that tetD, tetG, tetX, intI1, tnpA-04, and tnpA-05 had higher relative abundance in original soils, and the relative abundance of most TRGs and MGEs declined after the microbial remediation. Network analysis indicated that tnpA may dominate the transfer of TRGs, and Massilia, Alkanibacter, Rhizomicrobium, Xanthomonadales, Acidobacteriaceae, and Xanthomonadaceae were possible hosts of TRGs or MGEs. This study comprehensively evaluated the effectiveness and the ecological effects of the TC-degrading bacterial consortium in soil environment.

Long-term alkaline conditions inhibit the relative abundances of tetracycline resistance genes in saline 4-chlorophenol wastewater treatment

Considering the occurrence and spread of antibiotic resistance genes (ARGs) pose significant risks to public health, the effects of long-term exposure to alkaline conditions on the relative abundances of tetracycline resistance genes (TRGs) were studied in saline 4-chlorophenol (4-CP) wastewater treatment. Alkaline conditions were maintained by supplying the co-metabolic carbon source of sodium acetate. Results showed that except for the 4-CP, the removal of pollutants was significantly inhibited, and the relative abundances of the most TRGs were repressed. In addition, the removal of pollutants and the relative abundances of TRGs were moderately affected by the NaCl addition. The proteins in the extracellular polymeric substances (EPS) played key roles in reducing the relative abundances of TRGs, which were altered by the microbial diversity. In conclusion, for the pollutants removal and ARGs reduction in refractory industrial wastewater treatment, alkaline conditions should be maintained by selecting suitable co-metabolic carbon sources.

Dissemination of resistance genes in duck/fish polyculture ponds in Guangdong Province: correlations between Cu and Zn and antibiotic resistance genes.

Duck/fish polyculture farming is a typical farming model in the Pearl River delta in southern China. We examined soil, water, and sediment samples from three duck-fish farms in Guangdong Province in September and December 2014. We determined the abundance of three metal resistance genes, 16S rDNA, and 23 antibiotic resistance genes encoding resistance to tetracycline, sulfonamides, quinolones, chloramphenicol, and β-lactamases. Microbial community structure was quantified by Illumina high-throughput sequencing of 16S rDNA genes. We found a prevalence of antibiotic resistance genes and the sul1, sul2, tetA, tetM, aac(6')-Ib, and floR genes were the most abundant. Levels of Cu and Zn were significantly correlated with numerous ARG types and sul2, floR, and tetM were identified as potential antibiotic resistance gene indicators. Cu levels were significantly and positively correlated with the relative abundance of sul3, tetT, tetW, qnrB, qnrS, fexB, sul1, sul2, tetM, and qnrA. Zn was significantly correlated to relative abundance of sul2, sul3, tetM, tetA, tetT, tetW, qnrA, qnrB, qnrS, aac(6')-Ib, qepA, blaSHV, cmlA, floR, fexA, cfr, and fexB. The levels of Acinetobacter, Brevibacillus, and Wautersiella showed significant positive correlations with metal resistance genes as well as qnrB, oqxA, oqxB, and blaSHV (p > 0.8). Sphingobacterium, Flavobacterium, Acidothermus, and Corynebacterium had significant correlations with abundance of tetracycline resistance genes, sulfonamide resistance genes, blaTEM, blaCTX, and cfr (p > 0.8). Sphingobacterium, Flavobacterium, Acidothermus, and Corynebacterium were most abundant in soil samples while Acinetobacter, Brevibacillus, and Wautersiella were most abundant in water samples. Dissemination of antibiotic resistance genes in aquaculture environments is extensive and tracing their origins is necessary to establish risk assessment methods required for aquatic environmental protection.

Abundances of Tetracycline Resistance Genes and Tetracycline Antibiotics during Anaerobic Digestion of Swine Waste.

The impact of anaerobic digestion of animal waste on the persistence of antibiotic resistance genes (ARGs) and antibiotics is not widely studied. Two identical, 800-L digesters seeded with swine slurry were followed up to 100 d in three separate trials. The trials received varying amounts of antibiotic-free corn ( L.) mixed with water to maintain the digestion process. Biogas production, seven tetracycline resistance () genes, and three tetracyclines and their transformation products were measured. Biogas production proportionally increased as the feeding loads increased between trials. In Trial 1, log gene copies showed small but statistically significant ( < 0.01) increases during digestion. In Trial 2, anaerobic digestion did not have a significant ( > 0.05) effect except for significant reductions in B ( < 0.0001) and G ( = 0.0335) log gene copies. In Trial 3, which received the highest amount of corn mix, log copies of the 16S ribosomal RNA and the genes significantly ( < 0.0001) reduced over time during digestion. Up to 36 μg L tetracycline, 112 μg L chlortetracycline, 11.9 mg L isochlortetracycline, and 30 μg L 4-epitetracycline were detected both in the liquid and solid digestates. Results of this study revealed that although anaerobic digestion of swine waste can produce useful biogas, it does not result in complete removal of bacteria, ARGs, and antibiotics regardless of differences in the feeding loads between trials. Further effluent and sludge treatments are required prior to their downstream use in crop production to minimize emergence and environmental dissemination of antimicrobial-resistant bacteria through animal manure.