Antibiotic resistance profile of facultative deep-sea psychro-piezophile bacteria from the Arabian Sea and their relation with physicochemical factors. - Related Documents




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308401.0000Antibiotic resistance profile of facultative deep-sea psychro-piezophile bacteria from the Arabian Sea and their relation with physicochemical factors. Antibiotic resistance (ABR) is a significant global challenge, with antibiotics from various sources ending up in the ocean and affecting marine life. Profiling ABR in deep-sea bacteria is crucial for understanding the spread of ABR from environmental microbes to clinical pathogen and vice-versa. We evaluated facultative psychro-piezophile deep-sea bacteria from different depths of the Arabian Sea for their resistance to 20 commercial antibiotics. Bacteria from Zone 5 (2000-3000 m) exhibited the highest multiple antibiotic resistance (MAR) index (0.90), identifying it as a significant reservoir of ABR. Zone 1 (5-100 m) isolates (average 20 %) showed the highest resistance to synthetic antibiotics. Zone 3 (500-1000 m) isolates were highly resistant to diverse classes of antibiotics, separating upper (zone 1 and 2 (100-500 m) and deeper sea zones (zone 4 (1000-2000 m) and 5). The identified isolates belong to Bacillus, Niallia, Escherichia, Cytobacillus, and Pseudomonas genera. Additionally, antibiotic resistance genes (ARGs) such as StrB (2 isolates) and SXT integrase (1 isolate) were detected only in Zone 5 isolates. The SulII gene (19 isolates) was present across all zones. PCA analysis revealed a negative correlation between resistance and physicochemical factors (macronutrients like phosphate (PO(4)(3-)), nitrate (NO(3)(-)), nitrite (NO(2)(-)), and ammonia (NH(3)); micronutrient and heavy metals like (iron (Fe), manganese (Mn), zinc (Zn), copper (Cu), nickel (Ni)), aluminium (Al), cadmium (Cd), and chromium (Cr)), except for Phosphate (0.65). Overall, this study is the first to provide valuable insights into the prevalence of ABR using culture-dependent methods and its correlation with physicochemical factors in the deep-sea environments of the Arabian Sea.202540088632
308610.9997Seasonal variation, virulence gene and antibiotic resistance of Vibrio in a semi-enclosed bay with mariculture (Dongshan Bay, Southern China). In this study, the virulence genes, antibiotic resistance of culturable Vibrio and the environmental factors affecting Vibrio abundance were analyzed in four seasons in DongShan Bay with different intensity of aquaculture practice. A total of 253 bacteria isolates were obtained, of which 177 Vibrio strains belonged to 26 species. Annual Vibrio abundance in this region ranged from 20 to 11,600 CFU mL(-1) and the most significant positive correlation occurred with temperature. Detection of 9 different Vibrio virulence genes revealed that most isolates contained atypical virulence genes in addition to the typical ones. In particular, virulence genes of hemolysin such as tdh, trh, and hlyA (6.32 %, 15.52 %, and 11.30 %) showed different degrees of horizontal gene transfer (HGT). In our antibiotic resistance test, the multiple antibiotic resistance (MAR) index of the isolates ranged from 0.01 to 0.03 in different seasons, and three MAR Vibrio strains were detected. Overall, our study sheds new light on the spatial distribution patterns and the occurrence of virulence genes and antibiotics resistance Vibrio isolated from a subtropical bay with intensive aquaculture. Our study provides a suitable microbial quality surveillance in a mariculture impacted coastal environment. It will help to establish effective disease prevention measures in this area and provide useful guidance and support for formulating local antibiotics use policies.202236113173
308920.9997Distribution characteristics of antibiotic resistance bacteria and related genes in urban recreational lakes replenished by different supplementary water source. The distribution characteristics of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in urban recreational water from different water-supply sources might be different. In this study, water samples were collected to detect the antibiotic resistance of heterotrophic bacteria to five antibiotics, and the content, phenotype, gene type and species distribution of resistant bacteria were analyzed. The results showed that the changes of bacteria resistance rate in two lakes to five kinds of antibiotics were synchronous with time, and it would reach its maximum in autumn. The detection of ARGs and int I in 80 resistance strains showed that the detection rate of tetG, tetA and int I was high. Here, 51.25% of the bacteria were doubly resistant to AMP-CTX. The 80 isolate strains were of nine genera and 19 species, among which Bacillus cereus, Escherichia coli, Aeromonas veronii, Aeromonas caviae and Raoultella ornithinolytica were the common ARB species in two lakes. Correlation analysis showed that the water temperature was significantly correlated with the content of ARB in sulfamethoxazole (SMZ) and cefotaxime (CTX) (p < 0.05), and the total phosphorus (TP) in FQ lake was significantly correlated with the content of AMP-resistant bacteria (p < 0.05), while there were no other correlations between the changes of other water quality indexes and the content of ARB (p > 0.05).202235228362
537330.9997Impact of soil supplemented with pig manure on the abundance of antibiotic resistant bacteria and their associated genes. This study was conducted to evaluate the abundance of antibiotic resistant bacteria and their resistance genes from agriculture soil supplemented with pig manure. Uncultivable soil sample was supplemented with pig manure samples under microcosm experimental conditions and plated on Luria-Bertani (LB) agar incorporated with commercial antibiotics. The supplementation of soil with 15% pig manure resulted in the highest increase in the population of antibiotic resistant bacteria (ARB)/multiple antibiotic resistant bacteria (MARB). Seven genera that included Pseudomonas, Escherichia, Providencia, Salmonella, Bacillus, Alcaligenes and Paenalcaligenes were the cultivable ARB identified. A total of ten antibiotic resistant bacteria genes (ARGs) frequently used in clinical or veterinary settings and two mobile genetic elements (MGEs) (Class 1 and Class 2 integrons) were detected. Eight heavy metal, copper, cadmium, chromium, manganese, lead, zinc, iron, and cobalt were found in all of the manure samples at different concentrations. Tetracycline resistance genes were widely distributed with prevalence of 50%, while aminoglycoside and quinolone-resistance gene had 16% and 13%, respectively. Eighteen ARB isolates carried more than two ARGs in their genome. Class 1 integron was detected among all the 18 ARB with prevalence of 90-100%, while Class 2 integron was detected among 11 ARB. The two classes of integron were found among 10 ARB. Undoubtedly, pig manure collected from farms in Akure metropolis are rich in ARB and their abundance might play a vital role in the dissemination of resistance genes among clinically-relevant pathogens.202337308603
308740.9997Diversity and abundance of antibiotic resistance of bacteria during the seedling period in marine fish cage-culture areas of Hainan, China. Antibiotic resistance has become an important focus of research in the aquaculture environment. However, few studies have evaluated antibiotic resistance during the seedling period in marine fish cage-culture areas. In this study, culture-dependent methods and quantitative polymerase chain reaction were used to identify and detect cultivable heterotrophic antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs), respectively, during the seedling period in a marine fish cage-culture areas of Hainan, China. Bacterial resistance to amoxicillin, erythromycin, and gentamicin was generally high (average on 27.67%, 23.61% and 37.32%, respectively), whereas resistance to furazolidone and nitrofurantoin was generally low (average on 0.14% and 7.425%). Alteromonas (32.72%) and Vibrio (24.77%) were the dominant genus of ARB. Most ARB were opportunistic pathogens, belonging to the phylum Proteobacteria (96.02%). The abundance of sul family genes was higher than that of tet family genes. Overall, the abundance of ARGs and the resistance rates in HW was highest.201930955742
308550.9996Urbanization led to the abundance of Gram-negative, chemo-organo-heterotrophs, and antibiotic resistance genes in the downstream regions of the Ganga River water of India. The present investigation assesses the bacterial microbiome and antibiotic resistance genes (ARGs) of the river Ganga from Uttarakhand (upstream region; US group) and Uttar Pradesh (downstream region; DS group) regions using a 16S rRNA amplicon-based metagenomic approach. Gram-negative, aerobic, and chemo-organotrophic bacteria made up the majority of the bacterial genera during the overall analysis. Physicochemical analysis revealed a higher concentration of nitrate and phosphate in the downstream sites of the Ganga River. The prevalence of Gemmatimonas, Flavobacterium, Arenimonas, and Verrucomicrobia in the water of the DS region indicates a high organic load. Pseudomonas and Flavobacterium emerged as the most prevalent genera among the 35 significantly different shared genera (p-value < 0.05) in the US and DS regions, respectively. Overall antibiotic resistance analysis of the samples showed the dominance of β-lactam resistance (33.92%) followed by CAMP (cationic antimicrobial peptide) resistance (27.75%), and multidrug resistance (19.17%), vancomycin resistance (17.84%), and tetracycline resistance (0.77%). While comparing, the DS group exhibited a higher abundance of ARGs over the US group, where the CAMP resistance and β-lactam ARGs were dominant in the respective regions. The correlation (p-value < 0.05) analysis showed that most bacteria exhibit a significant correlation with tetracycline resistance followed by the phenicol antibiotic. The present findings draw attention to the need for regulated disposal of multiform human-derived wastes into the Ganga River to reduce the irrepressible ARGs dissemination.202337217817
308860.9996Occurrence of sulfonamide and tetracycline-resistant bacteria and resistance genes in aquaculture environment. The occurrence of sulfonamide and tetracycline resistance and their pollution profile in the aquaculture environment of Tianjin, northern China, were investigated. The presence of antibiotic-resistant bacteria was identified and the corresponding antibiotic resistance genes (ARGs) were quantified at 6 aquaculture farms in Tianjin. Sulfonamide-resistance genes were prevalent and their concentrations were the highest detected (3.0 × 10(-5) to 3.3 × 10(-4) for sul1/16S rDNA, 2.0 × 10(-4) to 1.8 × 10(-3) for sul2/16S rDNA) among the various ARGs, most likely because the use of sulfonamides is more prevalent than tetracyclines in this area. Bacillus was the most dominant bacterial genus in both sulfamethoxazole resistant bacteria (63.27% of the total resistant bacteria) and tetracycline-resistant bacteria (57.14% of the total resistant bacteria). At least two of those genes (tetM, tetO, tetT, tetW, sul1 and sul2) were detected in the isolates of Bacillus cereus, Bacillus subtilis, Bacillus megaterium and Acinetobacter lwofii, and all of the above genes were detected in B. cereus, suggesting the occurrence of multi-resistance in the studied area. The genetic transfer of sul1 between intestinal bacteria (e.g., Enterococcus spp.) and indigenous bacteria (e.g., Bacillus spp.) was implied by phylogenetic analysis. Several strains of resistant opportunistic pathogens (e.g., Acinetobacter spp.) were found in indigenous bacteria, which increase the risk of ARGs to public health. Overall, this is the first study to comprehensively investigate the antibiotic resistance profile by analyzing the species of antibiotic-resistant bacteria and adopting qualitative and quantitative methods to investigate ARGs at a typical aquaculture area in northern China.201222377146
337170.9996Ubiquitous and persistent Proteobacteria and other Gram-negative bacteria in drinking water. Drinking water comprises a complex microbiota, in part shaped by the disinfection and distribution systems. Gram-negative bacteria, mainly members of the phylum Proteobacteria, represent the most frequent bacteria in drinking water, and their ubiquity and physiological versatility raises questions about possible implications in human health. The first step to address this concern is the identification and characterization of such bacteria that is the first objective of this study, aiming at identifying ubiquitous or persistent Gram-negative bacteria, Proteobacteria or members of other phyla, isolated from tap water or from its source. >1000 bacterial isolates were characterized and identified, and a selected group (n=68) was further analyzed for the minimum inhibitory concentrations (MIC) to antibiotics (amoxicillin and gentamicin) and metals (copper and arsenite). Total DNA extracts of tap water were examined for the presence of putatively acquired antibiotic resistance or related genes (intI1, bla(TEM), qnrS and sul1). The ubiquitous tap water genera comprised Proteobacteria of the class Alpha- (Blastomonas, Brevundimonas, Methylobacterium, Sphingobium, Sphingomonas), Beta- (Acidovorax, Ralstonia) and Gamma- (Acinetobacter and Pseudomonas). Persistent species were members of genera such as Aeromonas, Enterobacter or Dechloromonas. Ralstonia spp. showed the highest MIC values to gentamicin and Acinetobacter spp. to arsenite. The genes intI1, bla(TEM) or sul1 were detected, at densities lower than 2.3×10(5)copies/L, 2.4×10(4)copies/L and 4.6×10(2)copies/L, respectively, in most tap water samples. The presence of some bacterial groups, in particular of Beta- or Gammaproteobacteria (e.g. Ralstonia, Acinetobacter, Pseudomonas) in drinking water may deserve attention given their potential as reservoirs or carriers of resistance or as opportunistic pathogens.201728238372
529080.9996Antibiotic resistance of bacteria isolated from heavy metal-polluted soils with different land uses. OBJECTIVES: The main objective of this study was to determine the relationship between the antibiotic and heavy metal tolerance of culturable bacteria isolated from mining waste, pasture, and agricultural soils containing different levels of heavy metals. MATERIALS AND METHODS: The populations of total culturable bacteria, and heavy metal- and antibiotic-tolerant bacteria in the soils were enumerated on nutrient agar, nutrient agar amended with metals, and Mueller-Hinton agar amended with antibiotics, respectively. The multiple antibiotic resistance index, and patterns of antibiotic resistance and heavy metal-antibiotic co-resistance were determined for 237 isolates. RESULTS: Among all the samples, those of the tailings of mines with higher levels of heavy metals had the lowest number of bacteria, but a relatively higher abundance of heavy metal- and antibiotic-resistant bacteria. A high degree of resistance was observed for ampicillin and amoxicillin in the isolates from all soils. The agricultural soil isolates had a high prevalence of resistance towards vancomycin, tetracycline, and streptomycin. Among all the tested antibiotics, gentamicin was the most potent. The most frequent pattern of multiple antibiotic resistance in the isolates from agricultural soils was amoxicillin, ampicillin, streptomycin, vancomycin, tetracycline, and doxycycline. The percentage of isolates with multiple antibiotic resistance was considerably higher in the agricultural soils than in the mining waste soils. A high rate of co-resistance towards Hg and antibiotics was observed among the gram-negative isolates, and towards Zn, Ni, Hg, and the beta-lactam antibiotics among the gram-positive isolates. CONCLUSIONS: The higher percentage of isolates with multiple antibiotic resistance in the agricultural soils that in the mining waste soils may be related to (1) the level of soil heavy metals, (2) the population and diversity of soil bacteria, (3) the application of manures, and (4) other factors affecting gene transfer between bacteria.201728732786
529890.9996Investigation of the antimicrobial susceptibility patterns of marine cyanobacteria in Bohai Bay: Cyanobacteria may be important hosts of antibiotic resistance genes in marine environment. Marine cyanobacteria, as widely distributed and photosynthetically autotrophic bacteria in the ocean, may contribute to the global dissemination of antibiotic resistance genes (ARGs) and develop a different antimicrobial susceptibility pattern from heterotrophic bacteria and cyanobacteria from freshwater environments. However, studies on antimicrobial susceptibility and the carriage of ARGs in marine cyanobacteria are still very limited. In this study, the antibiotic resistance characteristics of cyanobacteria in nearshore waters were examined through field monitoring and laboratory investigations, which included PCR detection and ARG transformation. The results showed a positive correlation between marine cyanobacteria and some ARGs in the nearshore waters of Bohai Bay. Moreover, most screened cyanobacteria showed high minimum inhibitory concentration (MIC) values for polymyxins, tetracyclines, kanamycin, and sulfonamides, moderate MIC values for streptomycin, chloramphenicol, rifampicin, and norfloxacin, and low MIC values for roxithromycin and cephalosporins. The bla(TEM), bla(KPC), sul1, sul2, strA, tetA, tetB, tetC, tetM, mdfA, and intI1 genes were detected in the screened marine cyanobacteria. The highest detection rates were observed for bla(TEM) (93.3 %), sul1 (56.6 %), sul2 (90 %), and strA (73.3 %). The detection rate of tetA (33.3 %) was the highest among the tetracycline resistance genes, and mdfA, a multidrug-resistant pump gene with resistance to tetracycline, also showed a high detection level (23.3 %). Overall, most of the screened marine cyanobacteria were found to tolerate multiple antibiotics in seawater, and the condition of the ARGs carriage was serious. Furthermore, the screened marine Synechocystis sp. C12-2 demonstrated the ability to accept ARGs on the RP4 plasmid through natural transformation and showed reduced sensitivity to ampicillin, suggesting the possibility that some marine cyanobacteria could acquire ARGs from the environment through horizontal gene transfer. Thus, marine cyanobacteria may play an important role in the propagation of marine ARGs.202437972772
2860100.9996Multi-drug resistance, integron and transposon-mediated gene transfer in heterotrophic bacteria from Penaeus vannamei and its culture environment. Multi-drug resistance (MDR) in bacteria is regarded as an emerging pollutant in different food production avenues including aquaculture. One hundred and sixty out of 2304 bacterial isolates from shrimp farm samples (n = 192) of Andhra Pradesh, India, were MDR. Based on biochemical identification and 16S rRNA sequencing, they were grouped into 35 bacterial species with the predominance of Vibrio parahaemolyticus (12.5%). The MDR isolates showed highest resistance toward oxytetracycline (89%) with more than 0.2 MAR (multiple antibiotic resistance), demonstrates a high-risk source. The most prevalent antibiotic-resistance gene (ARG) and mobile genetic element (MGE) detected were tetA (47.5%) and int1 (46.2%), respectively. In conjugation experiments, overall transfer frequency was found to be in the range of 1.1 × 10(-9) to 1.8 × 10(-3) with the transconjugants harbouring ARGs and MGEs. This study exposed the wide distribution of MDR bacteria in shrimp and its environment, which can further aggravate the already raised concerns of antibiotic residues in the absence of proper mitigation measures.202235066837
5317110.9996Effect of anaerobic digestion on pathogens and antimicrobial resistance in the sewage sludge. Antimicrobial resistance (AMR) is recognized as a global threat. AMR bacteria accumulate in sewage sludge however, knowledge on the persistence of human pathogens and AMR in the sludge line of the wastewater treatment is limited. Sludge can be used, with or without additional treatment, as fertilizer in agricultural fields. The aim of this study is to obtain knowledge about presence of human pathogens and AMR in the sewage sludge, before and after the anaerobic digestion (AD) applying innovative combinations of methods. Fifty sludge samples were collected. Cultivation methods combined with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and Antibiotic Susceptibility Test (AST) were used obtaining knowledge about the microbial community, pathogens, and antibiotic resistant bacteria while the droplet digital Polymerase Chain Reaction (ddPCR) was performed to detect most common AMR genes. In total, 231 different bacterial species were identified in the samples. The most abundant species were spore-forming facultative anaerobic bacteria belonging to Bacillus and Clostridium genera. The AD causes a shift in the microbial composition of the sludge (p = 0.04). Seven pathogenic bacterial species constituting 188 colonies were isolated and tested for susceptibility to Clindamycin, Meropenem, Norfloxacin, Penicillin G, and Tigecycline. Of the Clostridium perfringens and Bacillus cereus isolates 67 and 50 %, respectively, were resistant to Clindamycin. Two B. cereus and two C. perfringens isolates were also resistant to other antibiotics showing multidrug resistance. ARGs (bla(OXA), bla(TEM), ermB, qnrB, tet(A)-(W), sulI-II) were present at 7-8 Log gene copies/kg of sludge. AD is the main driver of a reduction of some ARGs (1 Log) but resistant bacteria were still present. The results showed the usefulness of the integration of the proposed analytical methods and suggest a decrease in the risk of presence of cultivable pathogens including resistant isolates after AD but a persistent risk of ARGs' horizontal transmission.202439244956
3090120.9996Networking and co-occurrence of virulent and multidrug resistant environmental bacteria in different aquatic systems: A gap in MDR-virulence transfer? Co-occurrence of resistance and virulence is often overlooked in aquatic bacteria as environmental reservoirs, while transmission of these characteristics to clinically significant strains present unforeseen problems in future. In this investigation, environmental bacteria identified concurrently from multiple aquatic habitats viz., groundwater, canal, river and coastal waters were profiled for antibiotic resistance, metal tolerance, virulence factors and genes coding for these determinants. Strains from polluted river and canal exhibited higher resistance and virulence, especially Pseudomonas gessardii and P. fluorescens displayed high antibiotic resistance index (ARI > 0.6-0.8) with Alkaline Protease and Phospholipase production. Opportunistic pathogens including Vibrio parahaemolyticus, V. alginolyticus, V. vulnificus, Corynebacterium and Comamonas testosteroni expressed all three virulence factors with relatively low resistance. However, V. vulnificus and V. alginolyticus exhibited multiclass antibiotic resistance (5/6 classes). Metagenomic analysis revealed that genes corresponding to beta-lactam resistance were significantly higher (p < 0.05) in freshwater than seawater, while multidrug resistance gene were higher (p < 0.05) in seawater. In all aquatic bodies, abundant virulence genes belonged to secretion system proteins followed by motility related genes. Culturable bacteria revealed differential distribution of positive and negative correlation between 31 targeted genes with expressed resistance and virulence. Among Acinetobacter, significant positive correlation was found between Phospholipase production, other virulence genes (OVGs) and resistance to DNA Synthesis Inhibitors (DSI). In Pseudomonas, positive correlation was detected between toxin genes (toxA, eta, hlyA and stx) and resistance to cell wall synthesis inhibitors (CSI) as well as with OVGs and adhesion genes (eae, afa, papC and papA). Network analysis displayed unique clustering of genes ncc, arsB, strA, merA and intI dominated by non-pathogens and distinct clustering of genes pho, erm, nfsA, trh, lasB, tdh and invA by Vibrio. This investigation extends insight on co-occurring resistance and virulence in aquatic reservoir bacteria that could pose serious threats to public health in future.202336206910
2867130.9996Enzymatic Activity and Horizontal Gene Transfer of Heavy Metals and Antibiotic Resistant Proteus vulgaris from Hospital Wastewater: An Insight. Globally, the issue of microbial resistance to medicines and heavy metals is getting worse. There are few reports or data available for Proteus vulgaris (P. vulgaris), particularly in India. This investigation intends to reveal the bacteria's ability to transmit genes and their level of resistance as well. The wastewater samples were taken from several hospitals in Lucknow City, India, and examined for the presence of Gram-negative bacteria that were resistant to antibiotics and heavy metals. The microbial population count in different hospital wastewaters decreases with increasing concentrations of metal and antibiotics. Among all the examined metals, Ni and Zn had the highest viable counts, whereas Hg, Cd, and Co had the lowest viable counts. Penicillin, ampicillin, and amoxicillin, among the antibiotics, demonstrated higher viable counts, whereas tetracycline and erythromycin exhibited lower viable counts. The MIC values for the P. vulgaris isolates tested ranged from 50 to 16,00 μg/ml for each metal tested. The multiple metal resistance (MMR) index, which ranged from 0.04 to 0.50, showed diverse heavy metal resistance patterns in all P. vulgaris isolates (in the case of 2-7 metals in various combinations). All of the tested isolates had methicillin resistance, whereas the least number of isolates had ofloxacin, gentamycin, or neomycin resistance. The P. vulgaris isolates displayed multidrug resistance patterns (2-12 drugs) in various antibiotic combinations. The MAR indexes were shown to be between (0.02-0.7). From the total isolates, 98%, 84%, and 80% had urease, gelatinase, and amylase activity, whereas 68% and 56% displayed protease and beta-lactamase activity. Plasmids were present in all the selected resistant isolates and varied in size from 42.5 to 57.0 kb and molecular weight from 27.2 to 37.0 MD. The transmission of the antibiotic/metal resistance genes was evaluated between a total of 7 pairs of isolates. A higher transfer frequency (4.4 × 10(-1)) was observed among antibiotics, although a lower transfer frequency (1.0 × 10(-2)) was observed against metals in both the media from the entire site tested. According to exponential decay, the population of hospital wastewater declined in the following order across all sites: Site II > Site IV > Site III > Site I for antibiotics and site IV > site II > site I >site III for metal. Different metal and antibiotic concentrations have varying effects on the population. The metal-tolerant P. vulgaris from hospital wastewater was studied in the current study had multiple distinct patterns of antibiotic resistance. It could provide cutting-edge methods for treating infectious diseases, which are essential for managing and assessing the risks associated with hospital wastewater, especially in the case of P. vulgaris.202236523753
2803140.9996Antimicrobial resistance, heavy metal resistance and integron content in bacteria isolated from a South African tilapia aquaculture system. Antibacterial compounds and metals co-select for antimicrobial resistance when bacteria harbour resistance genes towards both types of compounds, facilitating the proliferation and evolution of antimicrobial and heavy metal resistance. Antimicrobial and heavy metal resistance indices of 42 Gram-negative bacteria from a tilapia aquaculture system were determined to identify possible correlations between these phenotypes. Agar dilution assays were carried out to determine susceptibility to cadmium, copper, lead, mercury, chromate and zinc, while susceptibility to 21 antimicrobial agents was investigated by disk diffusion assays. Presence of merA, the mercury resistance gene, was determined by dot-blot hybridizations and PCR. Association of mercury resistance with integrons and transposon Tn21 was also investigated by PCR. Isolates displayed a high frequency of antimicrobial (erythromycin: 100%; ampicillin: 85%; trimethoprim: 78%) and heavy metal (Zn2+: 95%; Cd2+: 91%) resistance. No correlation was established between heavy metal and multiple antibiotic resistance indices. Significant positive correlations were observed between heavy metal resistance profiles, indices, Cu2+ and Cr3+ resistance with erythromycin resistance. Significant positive correlations were observed between merA (24%)/Tn21 (24%) presence and heavy metal resistance profiles and indices; however, significant negative correlations were obtained between integron-associated qacE∆1 (43%) and sulI (26%) gene presence and heavy metal resistance indices. Heavy metal and antimicrobial agents co-select for resistance, with fish-associated, resistant bacteria demonstrating simultaneous heavy metal resistance. Thus, care should be taken when using anti-fouling heavy metals as feed additives in aquaculture facilities.201729160218
5318150.9996Further Evidence of Anthropogenic Impact: High Levels of Multiple-Antimicrobial-Resistant Bacteria Found in Neritic-Stage Sea Turtles. BACKGROUND/OBJECTIVES: Marine turtles are globally threatened and face daily anthropogenic threats, including pollution. Water pollution from emerging contaminants such as antimicrobials is a major and current environmental concern. METHODS: This study investigated the phenotypic antimicrobial resistance and heavy metal resistance genes of 47 Vibrio isolates from different stages of sea turtles (oceanic stage vs neritic stage) from the Taiwanese coast. RESULTS: The results show that a high proportion (48.9%; 23/47) of the Vibrio species isolated from sea turtles in our study had a multiple antimicrobial resistance (MAR) pattern. It was found that Vibrio spp. isolates with a MAR pattern and those with a MAR index value greater than 0.2 were both more likely to be observed in neritic-stage sea turtles. Furthermore, isolates from neritic-stage sea turtles exhibited greater resistance to the majority of antimicrobials tested (with the exception of beta-lactams and macrolides) than isolates from the oceanic-stage groups. Isolates from neritic sea turtles were found to be more resistant to nitrofurans and aminoglycosides than isolates from oceanic sea turtles. Furthermore, isolates with a MAR pattern (p = 0.010) and those with a MAR index value greater than 0.2 (p = 0.027) were both found to be significantly positively associated with the mercury reductase (merA) gene. CONCLUSIONS: The findings of our study indicate that co-selection of heavy metals and antimicrobial resistance may occur in aquatic bacteria in the coastal foraging habitats of sea turtles in Taiwan.202439596693
7086160.9996Detection of faecal bacteria and antibiotic resistance genes in biofilms attached to plastics from human-impacted coastal areas. Plastics have been proposed as vectors of bacteria as they act as a substrate for biofilms. In this study, we evaluated the abundance of faecal and marine bacteria and antibiotic resistance genes (ARGs) from biofilms adhered to marine plastics. Floating plastics and plastics from sediments were collected in coastal areas impacted by human faecal pollution in the northwestern Mediterranean Sea. Culture and/or molecular methods were used to quantify faecal indicators (E. coli, Enterococci and crAssphage), and the ARGs sulI, tetW and bla(TEM) and the 16S rRNA were detected by qPCR assays. Pseudomonas and Vibrio species and heterotrophic marine bacteria were also analysed via culture-based methods. Results showed that, plastic particles covered by bacterial biofilms, primarily consisted of marine bacteria including Vibrio spp. Some floating plastics had a low concentration of viable E. coli and Enterococci (42% and 67% of the plastics respectively). Considering the median area of the plastics, we detected an average of 68 cfu E. coli per item, while a higher concentration of E. coli was detected on individual plastic items, when compared with 100 ml of the surrounding water. Using qPCR, we quantified higher values of faecal indicators which included inactive and dead microorganisms, detecting up to 2.6 × 10(2) gc mm(-2). The ARGs were detected in 67-88% of the floating plastics and in 29-57% of the sediment plastics with a concentration of up to 6.7 × 10(2) gc mm(-2). Furthermore, enrichment of these genes was observed in biofilms compared with the surrounding water. These results show that floating plastics act as a conduit for both the attachment and transport of faecal microorganisms. In contrast, low presence of faecal indicators was detected in plastic from seafloor sediments. Therefore, although in low concentrations, faecal bacteria, and potential pathogens, were identified in marine plastics, further suggesting plastics act as a reservoir of pathogens and ARGs.202336596379
5371170.9995Prevalence of antimicrobial resistance in a full-scale drinking water treatment plant. Antibiotic resistance in drinking water has received increasing attention in recent years. In this study, the occurrence and abundance of antibiotic resistance genes (ARGs) in a drinking water treatment plant (DWTP) was comprehensively investigated using metagenomics. Bioinformatics analysis showed that 381 ARG subtypes belonging to 15 ARG types were detected, and bacitracin had the highest abundance (from 0.26 × 10(-2) to 0.86 copies/cell), followed by multidrug (from 0.57 × 10(-1) to 0.47 copies/cell) and sulfonamide (from 0.83 × 10(-2) to 0.35 copies/cell). Additionally, 933 ARG-carrying contigs (ACCs) were obtained from the metagenomic data, among which 153 contigs were annotated as pathogens. The most abundant putative ARG host was Staphylococcus (7.9%), which most frequently carried multidrug ARGs (43.2%). Additionally, 38 high-quality metagenome-assembled genomes (MAGs) were recovered, one of which was identified as Staphylococcus aureus (Bin.624) and harboured the largest number of ARGs (n = 16). Using the cultivation technique, 60 isolates were obtained from DWTP samples, and Staphylococcus spp. (n = 11) were found to be dominant in all isolates, followed by Bacillus spp. (n = 17). Antimicrobial susceptibility testing showed that most Staphylococcus spp. were multidrug resistant (MDR). These results deepen our understanding of the distribution profiles of ARGs and antibiotic resistant bacteria (ARB) in DWTPs for potential health risk evaluation. Our study also highlights the need for new and efficient water purification technologies that can be introduced and applied in DWTPs.202337331316
3373180.9995Evidence of Increased Antibiotic Resistance in Phylogenetically-Diverse Aeromonas Isolates from Semi-Intensive Fish Ponds Treated with Antibiotics. The genus Aeromonas is ubiquitous in aquatic environments encompassing a broad range of fish and human pathogens. Aeromonas strains are known for their enhanced capacity to acquire and exchange antibiotic resistance genes and therefore, are frequently targeted as indicator bacteria for monitoring antimicrobial resistance in aquatic environments. This study evaluated temporal trends in Aeromonas diversity and antibiotic resistance in two adjacent semi-intensive aquaculture facilities to ascertain the effects of antibiotic treatment on antimicrobial resistance. In the first facility, sulfadiazine-trimethoprim was added prophylactically to fingerling stocks and water column-associated Aeromonas were monitored periodically over an 11-month fish fattening cycle to assess temporal dynamics in taxonomy and antibiotic resistance. In the second facility, Aeromonas were isolated from fish skin ulcers sampled over a 3-year period and from pond water samples to assess associations between pathogenic strains to those in the water column. A total of 1200 Aeromonas isolates were initially screened for sulfadiazine resistance and further screened against five additional antimicrobials. In both facilities, strong correlations were observed between sulfadiazine resistance and trimethoprim and tetracycline resistances, whereas correlations between sulfadiazine resistance and ceftriaxone, gentamicin, and chloramphenicol resistances were low. Multidrug resistant strains as well as sul1, tetA, and intI1 gene-harboring strains were significantly higher in profiles sampled during the fish cycle than those isolated prior to stocking and these genes were extremely abundant in the pathogenic strains. Five phylogenetically distinct Aeromonas clusters were identified using partial rpoD gene sequence analysis. Interestingly, prior to fingerling stocking the diversity of water column strains was high, and representatives from all five clusters were identified, including an A. salmonicida cluster that harbored all characterized fish skin ulcer samples. Subsequent to stocking, diversity was much lower and most water column isolates in both facilities segregated into an A. veronii-associated cluster. This study demonstrated a strong correlation between aquaculture, Aeromonas diversity and antibiotic resistance. It provides strong evidence for linkage between prophylactic and systemic use of antibiotics in aquaculture and the propagation of antibiotic resistance.201627965628
5336190.9995Resistant Genes and Multidrug-Resistant Bacteria in Wastewater: A Study of Their Transfer to the Water Reservoir in the Czech Republic. Wastewater is considered the most serious source of the spread of antibiotic resistance in the environment. This work, therefore, focuses on the fate and spread of antibiotic resistance genes (ARGs) in wastewater and the monitoring of multidrug-resistant strains. ARGs were monitored in the nitrification and sedimentation tanks of the wastewater treatment plant (WWTP) and in the dam into which this WWTP flows, at various times. The highest relative abundance was found for the blaTEM > tetW > blaNDM-1 > vanA resistance genes, respectively. An increased concentration of tetracycline (up to 96.00 ng/L) and ampicillin (up to 19.00 ng/L) was found in water samples compared to other antibiotics detected. The increased incidence of seven ARGs and four antibiotics was observed in the November and December sampling times. Isolated ampicillin-resistant strains showed a high degree of resistance to ampicillin (61.2% of the total isolates had a minimum inhibitory concentration (MIC) ≥ 20 mg/mL). In 87.8% of isolates, out of the total number, the occurrence of two or more ARGs was confirmed. These multidrug-resistant strains were most often identified as Aeromonas sp. This strain could represent a significant role in the spread of multidrug resistance through wastewater in the environment.202235207435