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Unveiling resistome profiles in the sediments of an Antarctic volcanic island
2019
Centurion, V.B. | Delforno, T.P. | Lacerda-Júnior, G.V. | Duarte, A.W.F. | Silva, L.J. | Bellini, G.B. | Rosa, L.H. | Oliveira, V.M.
The Deception Island, located in Maritime Antarctica, is a volcanic island with geothermal activity and one of the most visited by tourists. However, the extent of the anthropogenic impact remains largely unknown and the factors shaping the resistance/tolerance mechanisms in the microbiomes from Whalers Bay ecosystems have never been investigated. In this context, this study aimed to reveal the resistome profiles of Whalers Bay sediments and correlate them with environmental factors. Samples were collected at four sites during the summer 2014/2015 along a transect of 27.5 m in the Whalers Bay sediments. DNA isolated from sediment samples was sequenced using the Illumina HiSeq platform. Bioinformatic analyses allowed the assembly of contigs and scaffolds, prediction of ORFs, and taxonomic and functional annotation using NCBI RefSeq database and KEGG orthology, respectively. Microorganisms belonging to the genera Psychrobacter, Flavobacterium and Polaromonas were shown to dominate all sites, representing 60% of taxonomic annotation. Arsenic (As), copper (Cu) and iron (Fe) were the most abundant metal resistance/tolerance types found in the microbiomes. Beta-lactam was the most common class related to antibiotics resistance/tolerance, corroborating with previous environmental resistome studies. The acridine class was the most abundant amongst the biocide resistance/tolerances, related to antiseptic compounds. Results gathered in this study reveal a repertoire of resistance/tolerance classes to antibiotics and biocides unusually found in Antarctica. However, given the volcanic nature (heavy metals-rich region) of Deception Island soils, this putative impact must be viewed with caution.
Show more [+] Less [-]Fate of microbial pollutants and evolution of antibiotic resistance in three types of soil amended with swine slurry
2019
Sui, Qianwen | Zhang, Junya | Chen, Meixue | Wang, Rui | Wang, Yawei | Wei, Yuansong
Swine waste is a reservoir of microbial pollutants, including pathogens, antibiotic resistance genes (ARGs) and antibiotic resistant bacteria (ARB); therefore, soil fertilized with swine waste is an essential pathway for the dissemination of microbial pollutants from concentrated swine farms to the public. To rationalize the intervals of swine wastes application and investigate the effects of soil type on the occurrences of microbial pollutants and antibiotic resistance, pot experiments were conducted with three typical soils, humic acrisol, calcaric cambisols and histosols, being collected from south, northwest and northeast China (soil-R, soil-Y and soil-B, respectively). The soils were amended with swine slurry, digestate and chemical fertilizers and then conducted for 172 days. The influence of microbial pollutants and antibiotic resistance in soil posed by digestate application was similar to that of the chemical fertilizers, while swine slurry posed high risks to the soil. Soil-B which had the highest organic matter and neutral pH was least influenced by the swine slurry amendment. tetG, tetM and ermF were persistent ARGs in the slurry treated soil, and their decay rates fitted to first-order kinetics in the order soil-B> soil-Y > soil-R. Putative pathogens showed strong correlations with ARGs, suggesting a risk of dissemination. The initial 43–82 days was the active phase of microbial pollution in slurry treated soil, during which time heavy metals, moisture content, total organic carbon and the microbial community were key factors contributing to changes in antibiotic resistance. Fertilization intervals of livestock wastes should be lengthened over the ARG active phase.
Show more [+] Less [-]Antibiotic resistance and microbiota in the gut of Chinese four major freshwater carp from retail markets
2019
Yuan, Li | Wang, Li | Li, Zheng-Hao | Zhang, Ming-Qi | Shao, Wei | Sheng, Guo-Ping
Fish-associated antibiotic resistance genes (ARGs) have attracted increasing attention due to their potential risks to human beings via the food chain. However, data are scarce regarding the antibiotic resistance in fish themselves. Herein, the antibiotic resistance genes (ARGs) were assessed in the gut of four major Chinese freshwater carp (i.e., silver carp, grass carp, bighead carp, and crucian carp) from food retail markets. Results show that the abundances of target ARGs (e.g., tetA, tetO, tetQ, tetW, sulI, sulII, and blaTEM₋₁) and class 1 integrase (intI1) were in the range 9.4 × 10⁻⁶ - 1.6 × 10⁻¹ and 6.7 × 10⁻⁵ - 5.2 × 10⁻² gene copies per 16S rRNA gene, respectively. The sulI, sulII, and tetQ strongly correlated with silver and mercury resistance genes (e.g., silE and merR). The microbial taxa of fish gut could be partly separated among retail markets based on the PCA analysis. About 15.0% of the OTUs in fish gut were shared and 74.5% of the shared OTUs were identified as Acidobacteria, Bacteroidetes, Chloroflexi, Cyanobacteria, Firmicutes, and Proteobacteria. These phyla may constitute the core microbiota in the guts of the four Chinese freshwater carp. The possible ARG hosts were revealed based on the network analysis, and the presence of pathogen-associated resistant genera in fish gut highlights the need to fully understand their potential human health risks.
Show more [+] Less [-]Occurrence of enterococci harbouring clinically important antibiotic resistance genes in the aquatic environment in Gauteng, South Africa
2019
Hamiwe, Thabo | Kock, Marleen M. | Magwira, Cliff A. | Antiabong, John F. | Ehlers, Marthie M.
The development of antibiotic resistance and dissemination of its determinants is an emerging public health problem as it compromises treatment options of infections that were, until recently, treatable. Investigation of outbreaks of vancomycin resistant enterococci (VRE) suggests that the environment serves as a significant reservoir for antibiotic resistance genes (ARGs). However, there is a paucity of data regarding the presence of ARGs in the water sources in South Africa. In this study, water samples collected from wastewater treatment plants (WWTPs), surface water and hospital sewage were screened for enterococci harbouring genes conferring resistance to four classes of antibiotics. Enterococci isolates harbouring ARGs were detected in raw influent and treated wastewater discharge from WWTPs and hospital sewage water. Plasmid and transposon encoded ermB (macrolide), tetM and tetL (tetracycline) as well as aph(3’)-IIIa (aminoglycosides) genes were frequently detected among the isolates, especially in E. faecalis. The presence of enterococci harbouring ARGs in the treated wastewater suggest that ARGs are discharged into the environment where their proliferation could be perpetuated. Among the enterococci clonal complexes (CCs) recovered from wastewater were E. faecium CC17 (ST18), which is frequently associated with hospital outbreaks and a novel E. faecalis sequence type (ST), ST780.
Show more [+] Less [-]Spatial and temporal variation of antibiotic resistance in marine fish cage-culture area of Guangdong, China
2019
Wu, Jinjun | Su, Youlu | Deng, Yiqin | Guo, Zhixun | Cheng, Changhong | Ma, Hongling | Liu, Guangfeng | Xu, Liwen | Feng, Juan
The rapid emergence and dissemination of antibiotic resistance poses a threat to human health and to the marine environment. We have investigated the abundance and diversity of antibiotic resistance genes (ARGs) and of antibiotic-resistant bacteria (ARB), during the seedling period, rearing period, and harvesting period in seven marine fish cage-culture areas in Guangdong. Spatial and temporal variations of AGRs and ARB were also analyzed. Culture-based methods and quantitative PCR were used to detect ARB and ARGs. Bacterial resistance rates were no significantly different within farming periods. The proportion of antibiotic-resistant bacteria was extremely low (average on 1.15%), except for oxytetracycline-resistant bacteria (average on 34.15%). Vibrio was the most common ARB. Sul1, tetB, and ermB, had the highest relative abundance. The abundance of ARGs in the harvesting period was significant highest. The total abundance of ARGs was highest at Raoping and lowest at Dayawan and Liusha. Most ARGs were associated with opportunistic pathogens. The environmental factors effecting ARB and ARGs are complex, and no key factors were identified. This study provides a theoretical basis for assessing the harmfulness of ARGs and ARB to food safety and human health.
Show more [+] Less [-]Dissemination of extended-spectrum β-lactamase-producing Escherichia coli carrying mcr-1 among multiple environmental sources in rural China and associated risk to human health
2019
Ji, Xiang | Zheng, Beiwen | Berglund, Björn | Zou, Huiyun | Sun, Qiang | Chi, Xiaohui | Ottoson, Jakob | Li, Xuewen | Lundborg, Cecilia Stålsby | Nilsson, Lennart E.
Antibiotic resistance among gram-negative bacteria is increasingly becoming a problem of global concern. Particularly problematic is the emergence of resistance to last-resort antibiotics such as carbapenems and colistin. The increasing number of reports on the plasmid-mediated colistin resistance gene mcr-1 in isolates worldwide is raising concerns for the future usefulness of this class of antibiotics. Dissemination of mcr-1 is believed to have originated mainly from animal breeding, however, the role of the environment as a transmission source is not yet fully understood. In the current study, 89 extended-spectrum β-lactamase-producing Escherichia coli isolated from 231 samples from different environmental sources in 12 villages in a rural area of Shandong, China, were screened for mcr-1. 17 (19.1%) mcr-1-positive isolates were found from different environmental sources, aggregated in 6 villages. Plasmids of three different Inc-groups carrying mcr-1 were confirmed, indicating that the widespread geographical distribution of mcr-1 in the local area is due to a number of different plasmids. Additionally, almost a third (29.4%) of the isolates carried virulence factors associated to intestinal pathogenic E. coli. These results illustrate the high complexity of the transmission patterns of mcr-1 among different environmental matrices on a local scale and the potential for the environment to facilitate dissemination and emergence of antibiotic-resistant and virulent strains of bacteria.
Show more [+] Less [-]Antibiotic resistance genes and bacterial communities in cornfield and pasture soils receiving swine and dairy manures
2019
Chen, Zeyou | Zhang, Wei | Yang, Luxi | Stedtfeld, Robert D. | Peng, Anping | Gu, Cheng | Boyd, Stephen A. | Li, Hui
Land application of animal manure could change the profiles of antibiotic resistant bacteria (ARB), antibiotic resistance genes (ARGs) and bacterial communities in receiving soils. Using high-throughput real-time quantitative PCR and 16S rRNA amplicon sequencing techniques, this study investigated the ARGs and bacterial communities in field soils under various crop (corn and pasture) and manure (swine and dairy) managements, which were compared with those of two non-manured reference soils from adjacent golf course and grassland. In total 89 unique ARG subtypes were found in the soil samples and they conferred resistance via efflux pump, cellular protection and antibiotic deactivation. Compared to the ARGs in the golf course and grassland soils (28 and 34 subtypes respectively), manured soils generally had greater ARG diversity (36–55 subtypes). Cornfield soil frequently receiving raw swine manure had the greatest ARG abundance. The short-term (one week) application of composted and liquid swine manures increased the diversity and total abundance of ARGs in cornfield soils. Intriguingly the composted swine manure only marginally increased the total abundance of ARGs, but substantially increased the number of ARG subtypes in the cornfield soils. The network analysis revealed three major network modules in the co-occurrence patterns of ARG subtypes, and the hubs of these major modules (intl1-1, vanC, and pncA) may be candidates for selecting indicator genes for surveillance of ARGs in manured soils. The network analyses between ARGs and bacteria taxa revealed the potential host bacteria for the detected ARGs (e.g., aminoglycoside resistance gene aacC4 may be mainly carried by Acidobacteriaceae). Overall, this study highlighted the potentially varying impact of various manure management on antibiotic resistome and microbiome in cornfield and pasture soils.
Show more [+] Less [-]Freshwater environments as reservoirs of antibiotic resistant bacteria and their role in the dissemination of antibiotic resistance genes
2019
Nnadozie, Chika F. | Odume, Oghenekaro Nelson
Freshwater environments are susceptible to possible contamination by residual antibiotics that are released through different sources, such as agricultural runoffs, sewage discharges and leaching from nearby farms. Freshwater environment can thus become reservoirs where an antibiotic impact microorganisms, and is an important public health concern. Degradation and dilution processes are fundamental for predicting the actual risk of antibiotic resistance dissemination from freshwater reservoirs. This study reviews major approaches for detecting and quantifying antibiotic resistance bacteria (ARBs) and genes (ARGs) in freshwater and their prevalence in these environments. Finally, the role of dilution, degradation, transmission and the persistence and fate of ARB/ARG in these environments are also reviewed. Culture-based single strain approaches and molecular techniques that include polymerase chain reaction (PCR), quantitative polymerase chain reaction (qPCR) and metagenomics are techniques for quantifying ARB and ARGs in freshwater environments. The level of ARBs is extremely high in most of the river systems (up to 98% of the total detected bacteria), followed by lakes (up to 77% of the total detected bacteria), compared to dam, pond, and spring (<1%). Of most concern is the occurrence of extended-spectrum β-lactamase producing Enterobacteriaceae, methicillin resistant Staphylococcus aureus (MRSA) and vancomycin resistant Enterococcus (VRE), which cause highly epidemic infections. Dilution and natural degradation do not completely eradicate ARBs and ARGs in the freshwater environment. Even if the ARBs in freshwater are effectively inactivated by sunlight, their ARG-containing DNA can still be intact and capable of transferring resistance to non-resistant strains. Antibiotic resistance persists and is preserved in freshwater bodies polluted with high concentrations of antibiotics. Direct transmission of indigenous freshwater ARBs to humans as well as their transitory insertion in the microbiota can occur. These findings are disturbing especially for people that rely on freshwater resources for drinking, crop irrigation, and food in form of fish.
Show more [+] Less [-]Hysterical tetracycline in intensive poultry farms accountable for substantial gene resistance, health and ecological risk in Egypt- manure and fish
2019
Mahmoud, Manal A.M. | Abdel-Mohsein, Hosnia S.
Although the poultry production sector plays a key role in sustaining the majority of animal protein demand in Egypt, the deleterious effects of widespread antibiotic resistance on health and environment are currently not well recognized. Litter and dropping samples from broiler and layer poultry farms as well as, tilapia samples from the Nile River and aquaculture farms were collected from Upper Egypt. Samples were extracted and examined for tetracycline residues [tetracycline (TC), chlortetracycline (CTC), oxytetracycline (OTC) and doxycycline (DC)] using HPLC. In addition, tetracycline resistance genes [tet (M), tet (W), tet (Q) and tet (G)] were screened from pooled intestinal contents collected from twelve broiler farms in Upper Egypt. The antibiotic resistance genes results revealed that tet (W) was confirmed to be expressed in all intestinal samples. In contrast, tet (Q) and tet (M) were detected only in 42% and 17% of the samples, respectively. CTC and OTC were the antimicrobial compounds with the highest concentrations in poultry litter and droppings, with concentrations of 6.05 and 2.47 μg g−1 (CTC) and 5.9 and 1.33 μg g−1 (OTC), respectively. However, the concentrations of DC were significantly higher than those of the other compounds in both aquaculture and Nile River tilapia. The tetracycline residue levels in aquaculture tilapia were significantly higher than those in Nile River tilapia. The hazard quotients (HQs) exceeded 1 for OTC, CTC and DC, which highlights the great risk of using broiler litter to fertilize agricultural land. Moreover, the presence of DC and CTC indicates that consumption of aquaculture tilapia poses a considerable health risk. Therefore, poultry litter or droppings containing tetracycline residues and tet resistance determinants used for aquaculture or as farmland fertilizers could be major sources of antibiotic resistance in fish, humans and environment.
Show more [+] Less [-]Expression of resistance genes instead of gene abundance are correlated with trace levels of antibiotics in urban surface waters
2019
Yi, Xinzhu | Lin, Chenghui | Ong, Eugene Jie Li | Wang, Mian | Li, Bolin | Zhou, Zhi
In this study, antibiotic resistance to macrolide-lincosamide-streptogramin B (MLSB) antibiotics in total microbial community in surface water in a coastal urban city was measured using a modified fluorescence in situ hybridization (FISH) technique. This FISH technique quantified the rate of antibiotic resistance to MLSB antibiotics through targeting methylation site of A2058 of 23S rRNAs resulting from expressed erythromycin ribosome methylation (erm) genes. Correlations between the rates of MLSB resistance measured by FISH and macrolide concentrations was stronger than that between the relative abundance of erm genes and macrolide concentrations, especially in residential areas where the main detected antibiotics were macrolides. These results suggest that trace levels of antibiotics in environmental waters, which was as low as 40 ng L−1, may still play important roles in the development and spread of antibiotic resistance. Additionally, methylation as a result of erm gene expression, instead of erm gene abundance, was a better indicator of selective pressure of trace level macrolides. The rates of MLSB resistance varied significantly among land use types, suggesting that anthropogenic activities are important factors to select for erm gene expression in the environment. Microbial community analysis of representative surface water samples showed that relatively high rates of MLSB resistance were observed in Alphaproteobacteria (42%), Acidobacteria (36%), Bacteroidaceae (32%), Chloroflexi (27%), and Betaproteobacteria (20.2%).
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