Search Results (26)

During intensive aquaculture of Eriocheir sinensis (E. sinensis), wastewater containing feed additives, heavy metals, and antibiotics can cause water eutrophication and lead to the accumulation of complex pollutants in sediments. In this study, heavy metals and antibiotic resistance genes (ARGs) were analyzed for their distribution and accumulation patterns via ICP-OES and qPCR, and their correlations with bacterial communities in sediments from E. sinensis ponds were evaluated using co-occurrence network and redundancy analysis. Our results demonstrated that aquaculture activities significantly elevated the concentrations of heavy metals such as Cadmium (Cd) and Manganese (Mn) in the sediments, and Cd was identified as the dominant factor contributing to the increase in the potential ecological risk index (RI). The abundance of ARGs significantly increased, with sulfonamide resistance genes (sul1, sul2), β-lactam resistance genes (blaTEM), and integron-associated genes (intl1) showing the highest levels. Correlation and redundancy analyses showed that most ARGs were positively correlated with and linked to Proteobacteria, Bacteroidetes, and Dechloromonas as potential hosts. Positive correlations were generally observed among heavy metals, suggesting a common source, namely aquaculture wastewater, and some showed positive associations with Acidobacteria. This study provides a theoretical basis for further understanding the distribution, accumulation patterns, and coupling relationships of heavy metals and ARGs, as well as their relation and effects on bacterial communities in the sediment of E. sinensis ponds. Full article

The growing and unprecedented water crisis leads to the need to find alternative water resources, and the reuse of treated urban wastewater is an excellent approach. Accordingly, in this work, the disinfection of a secondary effluent (W) discharged from a wastewater treatment plant (WWTP) by hydrogen peroxide combined with radiation (H₂O₂+UV/visible) was studied with the aim of obtaining treated water that can be reused. Firstly, the effect of hydrogen peroxide alone, radiation per se and the combined H₂O₂+UV/Visible process in the inactivation of enterobacteria were assessed. It was found that the oxidant alone is not efficient; the maximum inactivation is achieved when the oxidant and radiation are used simultaneously. For the first time, the effect of some operational parameters, namely the hydrogen peroxide concentration (between 50 and 125 mg/L), initial pH (from 5.0 to 7.0), temperature (between 15 and 25 °C), and radiation intensity (100 to 500 W/m²), on the efficiency of the disinfection process was assessed. When the process was carried out under the best operating conditions found ([H₂O₂] = 75 mg/L, pH = 5.0, T = 25 °C, and UV/visible light with I = 500 W/m²), total enterobacteria and total heterotrophs were inactivated and the abundance of the 16S rRNA, bla_TEM, qnrS, and intl1 genes was reduced. The cultivable microorganisms grew again after 3 days of storing the treated wastewater (TW), making it impossible to reuse such effluent after storage. Therefore, the potential capacity of a diverse bacterial community present in river water to inhibit the regrowth of potentially harmful bacteria present in the urban secondary wastewater after the application of the treatment process was also evaluated. To the authors’ knowledge, this has never been studied before. For this purpose, the TW was diluted with river water (R) at a volumetric percentage of 50/50—sample R+TW. It was found that, after storage, only the total heterotrophs grew, while the abundance of the targeted genes remained practically constant. The R+TW sample after storage met the legal limits for reuse in urban and agricultural applications. The results of this study suggest that the combination of the H₂O₂+UV/visible radiation treatment with dilution of the final treated effluent with natural surface water can contribute to reducing the burden of water scarcity. Full article

The use of dairy slurries as organic fertilizer amendments is a common practice in agriculture as a cost-saving measure, as well as a residue management strategy. However, concerns related to the increase in antibiotic resistance in the environment under the scope of the One Health strategy are increasing. In this study, we aimed to assess resistome enrichment driven by dairy slurry application in four southern Chile dairy farms. Slurry pits, rhizospheres of Lolium perenne amended with those slurries, and bulk soils were sampled. Thirteen antibiotic-resistance genes (ARGs, tetA, tetG, tetM, tetQ, tetW, tetX, sul1, sul2, blaCTX_M, bla_OXA-1, blaTEM, ermB, and dfrA1) for five antibiotic classes (tetracyclines, sulfonamides, beta-lactams, macrolides, and trimethoprim–sulfamethoxazole), two related integrases (intl1 and intl2), and total bacteria (16S rRNA) abundance was measured by quantitative PCR (qPCR). Then, the abundance profiles of two enzyme-inactivated ARGs (tetX and blaTEM) were determined. The differences between the bacterial communities inhabiting the different sample types were explored with 16S rRNA metabarcoding. In general, all measured ARGs were detected in slurries. A decreasing trend in ARG copy numbers was observed with increasing soil depth, with the exception of tetX, whose abundance increased in the bulk soil at specific farms. The tetX and blaTEM communities revealed no differences in the relative abundance of variants in any of the samples. Finally, taxonomic and structural differences were found among all sample types. Thus, the enrichment of the sampled farm soil resistomes was driven by the application of the raw slurries as fertilizer. Full article

In 2019, a disease outbreak struck a hybrid sturgeon farm (Huso dauricus ♀ × A. schrenckii ♂) in Tiantai, Zhejiang province, leading to the deaths of 8000 sturgeons. The sturgeons exhibited reduced appetite, lethargic and uncoordinated swimming, and physical signs such as reddish petechiae and ulcers on the body and fins. Hemorrhagic spots were observed on the kidneys, spleen, and gonads, alongside reddish intestines with hemorrhagic ascites in the abdominal cavity. ST-1902 was isolated and identified as Aeromonas hydrophila through physiological and biochemical characterization and 16S rDNA sequence analysis. The pathogenicity of ST-1902 was confirmed through a challenge test, with a median lethal dosage (LD50) of 7.9 × 10⁶ CFU/IND. Histopathological examination showed hyperplasia and neoplasm-like changes in the epicedial mesothelial tissues, enlarged and necrosis renal tissue, and serious hemosiderosis in spleen and gills. Virulent genes (Aer, Epa, Alt, Hly, and Act) were detected in ST-1902, corresponding to typical β-hemolysis, extracellular protease, and enterotoxin. Moreover, antimicrobial experiment detection indicated ST-1902 is sensitive to quinolones and phenicols but resistant to sulfamethoxazole, aminoglycoside antibiotics with Sul1, and Intl and Ant (3”)-I. These results suggest that A. hydrophila was the causative agent of the sturgeon disease and highlight the emerging threat it poses to the sturgeon industry. Full article

Microbial resistance to antibiotics poses a significant threat to both human and animal health, necessitating international efforts to mitigate this issue. This study aimed to assess the resistance profiles of Salmonella sp. isolates and identify the presence of intl1, sul1, and blaTEM resistance genes within antigenically characterized isolates, including Agona, Livingstone, Cerro, Schwarzengrund, Salmonella enterica subsp. enterica serotype O:4.5, Anatum, Enteritidis, Johannesburg, Corvallis, and Senftenberg. These isolates underwent susceptibility testing against 14 antibiotics. The highest resistance percentages were noted for sulfamethoxazole (91%), sulfonamides (51%), and ceftiofur (28.9%), while no resistance was observed for ciprofloxacin. Salmonella Johannesburg and Salmonella Corvallis showed resistance to one antibiotic, whereas other serovars were resistant to at least two. Salmonella Schwarzengrund exhibited resistance to 13 antibiotics. The intl1 gene was detected in six out of the ten serovars, and the sul1 gene in three, always co-occurring with intl1. The blaTEM gene was not identified. Our findings highlight the risk posed by the detected multiple resistances and genes to animal, human, and environmental health. The multidrug resistance, especially to third-generation cephalosporins and fluoroquinolones, highlights the need for stringent monitoring of Salmonella in laying hens. The potential of the environment, humans, eggs, and their products to act as vectors for antibiotic resistance represents a significant concern for One Health. Full article

The role of marine environments in the global spread of antibiotic resistance still remains poorly understood, leaving gaps in the One Health-based research framework. Antibiotic resistance genes (ARGs) encoding resistance to five major antibiotic classes, including sulfonamides (sul1, sul2), tetracyclines (tetA, tetB), β-lactams (bla_CTX-M, bla_TEM bla_VIM), macrolides (ermB, mphA), aminoglycosides (aac3-2), and integrase gene (intl1) were quantified by RT-qPCR, and their distribution was investigated in relation to environmental parameters and the total bacterial community in bottom layer and surface waters of the central Adriatic (Mediterranean), over a 68 km line from the wastewater-impacted estuary to coastal and pristine open sea. Seasonal changes (higher in winter) were observed for antibiotic resistance frequency and the relative abundances of ARGs, which were generally higher in eutrophic coastal areas. In particular, intl1, followed by bla_TEM and bla_VIM, were strongly associated with anthropogenic influence and Gammaproteobacteria as their predominant carriers. Water column stratification and geographic location had a significant influence on ARGs distribution in the oligotrophic zone, where the bacterial community exhibited a seasonal shift from Gammaproteobacteria in winter to Marine group II in summer. Full article

Antibiotic-resistance gene elements (ARGEs) such as antibiotic-resistance genes (ARGs), integrons, and plasmids are key to the spread of antimicrobial resistance (AMR) in marine environments. Kuwait’s marine area is vulnerable to sewage contaminants introduced by numerous storm outlets and indiscriminate waste disposal near recreational beaches. Therefore, it has become a significant public health issue and warrants immediate investigation. Coliforms, especially Gram-negative Escherichia coli, have been regarded as significant indicators of recent fecal pollution and carriers of ARGEs. In this study, we applied a genome-based approach to identify ARGs’ prevalence in E. coli isolated from mollusks and coastal water samples collected in a previous study. In addition, we investigated the plasmids and intl1 (class 1 integron) genes coupled with the ARGs, mediating their spread within the Kuwait marine area. Whole-genome sequencing (WGS) identified genes resistant to the drug classes of beta-lactams (bla_CMY-150, bla_CMY-42, bla_CTX-M-15, bla_DHA-1, bla_MIR-1, bla_OKP-B-15, bla_OXA-1, bla_OXA-48, bla_TEM-1B, bla_TEM-35), trimethoprim (dfrA14, dfrA15, dfrA16, dfrA1, dfrA5, dfrA7), fluroquinolone (oqxA, oqxB, qnrB38, qnrB4, qnrS1), aminoglycoside (aadA2, ant(3’’)-Ia, aph(3’’)-Ib, aph(3’)-Ia, aph(6)-Id), fosfomycin (fosA7, fosA_6, fosA, fosB1), sulfonamide (sul1, sul2, sul3), tetracycline (tet-A, tet-B), and macrolide (mph-A). The MFS-type drug efflux gene mdf-A is also quite common in E. coli isolates (80%). The plasmid ColRNAI was also found to be prevalent in E. coli. The integron gene intI1 and gene cassettes (GC) were reported to be in 36% and 33%, respectively, of total E. coli isolates. A positive and significant (p < 0.001) correlation was observed between phenotypic AMR-intl1 (r = 0.311) and phenotypic AMR-GC (r = 0.188). These findings are useful for the surveillance of horizontal gene transfer of AMR in the marine environments of Kuwait. Full article

The low-efficiency rate of urban wastewater (UWW) treatment generates tons of discharged water with a high concentration of pollutants, pathogens and antibiotic-resistance genes (ARGs). Microalgal systems may be a green alternative to be implemented as a UWW polishing treatment. This study assessed the ability of Chlorella vulgaris and UWW autochthonous microalgal species (AMS) to simultaneously remove PO₄–P, and reduce the proliferation of coliforms and ARGs. AMS seems to be more promising due to: (i) the higher specific growth rate, μ_max (0.687 ± 0.065 d⁻¹); (ii) efficient PO₄–P removal (92.62 ± 0.10%); (iii) faster reduction of coliforms proliferation achieving concentrations below the limits of quantification (6 d); (iv) the reduction of intl1 and the ARGs sul1 and bla_TEM abundance in ca. of 70.4%, 69.2%, and 75.7%, respectively (9 d); and (v) the additional reduction of these genes in ca. of 97.1%, 94.2%, and 99.9%, respectively, after 5 d storage in the dark and at room temperature. Results also revealed that the high pH values in both microalgal systems (due to microalgal growth) were highly correlated with a reduction in the proliferation of coliforms, including Escherichia coli. In conclusion, using AMS as a final polishing treatment of UWW seems to be very promising. Full article

The aim of the study was phenotypic and genotypic characterization of Erysipelothrix rhusiopathiae strains isolated from diseased pigs in Poland and comparison of the SpaA (Surface protective antigen A) sequence of wild-type strains with the sequence of the R32E11 vaccine strain. The antibiotic susceptibility of the isolates was assessed using the broth microdilution method. Resistance genes, virulence genes, and serotype determinants were detected using PCR. The gyrA and spaA amplicons were sequenced to determine nonsynonymous mutations. The E. rhusiopathiae isolates (n = 14) represented serotypes 1b (42.8%), 2 (21.4%), 5 (14.3%), 6 (7.1%), 8 (7.1%), and N (7.1%). All strains were susceptible to β-lactams, macrolides and florfenicol. One isolate showed resistance to lincosamides and tiamulin, and most strains were resistant to tetracycline and enrofloxacin. High MIC values of gentamicin, kanamycin, neomycin, trimethoprim, trimethoprim/sulfadiazine, and rifampicin were recorded for all isolates. Phenotypic resistance was correlated with the presence of the tetM, int-Tn, lasE, and lnuB genes. Resistance to enrofloxacin was due to a mutation in the gyrA gene. All strains contained the spaA gene and several other genes putatively involved in pathogenesis (nanH.1, nanH.2, intl, sub, hlyA, fbpA, ERH_1356, cpsA, algI, rspA and rspB) Seven variants of the SpaA protein were found in the tested strains, and a relationship between the structure of SpaA and the serotype was noted. E. rhusiopathiae strains occurring in pigs in Poland are diverse in terms of serotype and SpaA variant and differ antigenically from the R32E11 vaccine strain. Beta-lactam antibiotics, macrolides, or phenicols should be the first choice for treatment of swine erysipelas in Poland. However, due to the small number of tested strains, this conclusion should be approached with caution. Full article

Pastures have become one of the most important sources of antibiotic resistance genes (ARGs) pollution, bringing risks to human health through the environment and the food that is grown there. Another significant source of food production is greenhouse horticulture, which is typically located near pastures. Through waterways, pasture-originated ARGs may transfer to the food in greenhouses. However, how these pasture-originated ARGs spread to nearby waterways and greenhouses has been much less investigated, while this may pose risks to humans through agricultural products. We analyzed 29 ARGs related to the most used antibiotics in livestock in the Netherlands at 16 locations in an agricultural area, representing pastures, greenhouses and lakes. We found that ARGs were prevalent in all surface waters surrounding pastures and greenhouses and showed a similar composition, with sulfonamide ARGs being dominant. This indicates that both pastures and greenhouses cause antibiotic resistance pressures on neighboring waters. However, lower pressures were found in relatively larger and isolated lakes, suggesting that a larger water body or a non-agricultural green buffer zone could help reducing ARG impacts from agricultural areas. We also observed a positive relationship between the concentrations of the class 1 integron (intl1 gene)—used as a proxy for horizontal gene transfer—and ARG concentration and composition. This supports that horizontal gene transfer might play a role in dispersing ARGs through landscapes. In contrast, none of the measured four abiotic factors (phosphate, nitrate, pH and dissolved oxygen) showed any impact on ARG concentrations. ARGs from different classes co-occurred, suggesting simultaneous use of different antibiotics. Our findings help to understand the spatial patterns of ARGs, specifically the impacts of ARGs from pastures and greenhouses on each other and on nearby waterways. In this way, this study guides management aiming at reducing ARGs′ risk to human health from agricultural products. Full article

Globally, cephalosporin therapy failure is a serious problem for infection control. One causative agent of cephalosporin-resistant infections is multidrug-resistant (MDR) E. coli producing extended-spectrum β-lactamases (ESBLs) and/or plasmid-encoded AmpC (pAmpC) β-lactamases. We evaluated the occurrence of ESBL/pAmpC genetic determinants in phenotypically MDR E. coli isolated from clinical samples of blood, faeces, ear effusion, urine and sputum from a UK hospital. Phenotypic resistance profiling for 18 antibiotics (from seven classes) showed that 32/35 isolates were MDR, with resistance to 4–16 of the tested antibiotics. Of the isolates, 97.1% showed resistance to ampicillin, 71.4% showed resistance to co-amoxiclav, cefotaxime, ceftazidime and ceftiofur, and 68.5% showed resistance to cefquinome. bla_CTX-M, bla_TEM and bla_OXA-1 genes were detected in 23, 13 and 12 strains, respectively, and Intl1 was detected in 17 isolates. The most common subtypes among the definite sequence types were CTX-M-15 (40%) and TEM-1 (75%). No E. coli isolates carried pAmpC genes. Significant correlations were seen between CTX-M carriage and cefotaxime, ceftiofur, aztreonam, ceftazidime and cefquinome resistance; between bla_CTX-M, bla_TEM and bla_OXA-1 carriage and ciprofloxacin resistance; and between Intl1 carriage and trimethoprim/sulfamethoxazole resistance. Thus, MDR phenotypes may be conferred by a relatively small number of genes. The level and pattern of antibiotic resistance highlight the need for better antibiotic therapy guidelines, including reduced use and improved surveillance. Full article

The application of livestock manure is the leading cause of antibiotic and heavy metal pollution in agricultural soil. However, the effects of oxytetracycline (OTC) and lead (Pb) pollution in the single or combined form on antibiotic resistance genes (ARGs) in the soil need to be further studied. This study was planned to investigate the effects of OTC and Pb application on ARGs, mobile genetic elements (MGEs), and bacterial abundance in the soil. The relative abundance of ARGs and MGEs increased by 0.31-fold and 0.03-fold after the addition of 80 mg kg⁻¹ Pb to the soil, and by 0.49-fold and 0.03-fold after the addition of 160 mg kg⁻¹ Pb. In addition, under the premise of the existence of OTC, the inhibitory effect of a low concentration of Pb on ARG is stronger than that of a high concentration of Pb, resulting in a lower abundance of ARGs. The abundance of ARGs and MGEs increased by 0.11-fold and 0.17-fold after the addition of OTC (30 mg kg⁻¹) to the soil at a Pb concentration of 80 mg kg⁻¹ and by 0.18-fold and 0.04-fold at a Pb concentration of 160 mg kg⁻¹. The addition of OTC and Pb in the soil also decreased the many bacterial communities such as Bacteroidetes, Proteobacteria, Acidobacteria, and Firmicutes. Redundancy analysis (RDA) showed that organic matter content and pH were positively correlated with the abundance of ARGs and MGEs. At the same time, electrical conductivity (EC) had a negative correlation with the abundance of ARGs and MGEs in the soil. Intl1 was significantly associated with tetB, sul1, tetQ, sul2, and sul3. Network analysis illustrated that Actinobacteria, Bacteroidetes, and Proteobacteria were the main host bacteria causing changes in the abundance of ARGs and MGEs, and they were also predominant phylum in the culture environment. This conclusion can provide a reference for the related research of ARGs in soil. Full article

SCUBA divers are predisposed to otitis externa caused by Pseudomonas aeruginosa, which is becoming increasingly multi-drug resistant (MDR). The present work assessed the antibiotic resistance profiles of P. aeruginosa obtained from SCUBA divers and their environment in Sodwana Bay, South Africa. Bacterial isolates from a total of 137 random water and ear swab samples were identified using biochemical and molecular methods. P. aeruginosa strains were further evaluated for antibiotic susceptibility using the Kirby–Bauer assay. Double disk synergy test (DDST) to confirm metallo-β-lactamase (MBL) production and PCR amplification of specific antibiotic resistance genes was performed. All (100%) 22 P. aeruginosa isolates recovered were resistant to 6 of the β-lactams tested including imipenem but exhibited susceptibility to trimethoprim–sulfamethoxazole. MBL production was observed in 77% of isolates while the most prevalent extended-spectrum β-lactamase (ESBL) genes present included bla_AmpC (86.9%) followed by bla_TEM (82.6%). Sulfonamide resistance was largely encoded by sul1 (63.6%) and sul2 (77.3%) genes with a high abundance of class 1 integrons (77.3%) of which 18.2% carried both Intl1 and Intl2. P. aeruginosa found in Sodwana Bay exhibits multi-drug resistance (MDRce) to several pharmaceutically important drugs with the potential to transfer antibiotic resistance to other bacteria if the judicious use of antibiotics for their treatment is not practiced. Full article

Chronic alcohol abuse causes an inflammatory response in the intestinal tract with damage to the integrity of the mucosa and epithelium, as well as dysbiosis in the gut microbiome. However, the role of gut bacteria in ethanol effects and how these microorganisms interact with the immune system are not well understood. The aim of the present study was to evaluate if TLR4 alters the ethanol-induced intestinal inflammatory response, and whether the response of this receptor affects the gut microbiota profile. We analyzed the 16S rRNA sequence of the fecal samples from wild-type (WT) and TLR4-knockout (TLR4-KO) mice with and without ethanol intake for 3 months. The results demonstrated that chronic ethanol consumption reduces microbiota diversity and causes dysbiosis in WT mice. Likewise, ethanol upregulates several inflammatory genes (IL-1β, iNOS, TNF-α) and miRNAs (miR-155-5p, miR-146a-5p) and alters structural and permeability genes (INTL1, CDH1, CFTR) in the colon of WT mice. Our results further demonstrated that TLR4-KO mice exhibit a different microbiota that can protect against the ethanol-induced activation of the immune system and colon integrity dysfunctions. In short, our results reveal that TLR4 is a key factor for determining the gut microbiota, which can participate in dysbiosis and the inflammatory response induced by alcohol consumption. Full article

Background: Uropathogenic Escherichia coli (UPEC) has increased the incidence of urinary tract infection (UTI). It is the cause of more than 80% of community-acquired cystitis cases and more than 70% of uncomplicated acute pyelonephritis cases. Aim: The present study describes the molecular epidemiology of UPEC O25b clinical strains based on their resistance profiles, virulence genes, and genetic diversity. Methods: Resistance profiles were identified using the Kirby–Bauer method, including the phenotypic production of extended-spectrum β-lactamases (ESBLs) and metallo-β-lactamases (MBLs). The UPEC serogroups, phylogenetic groups, virulence genes, and integrons were determined via multiplex PCR. Genetic diversity was established using pulsed-field gel electrophoresis (PFGE), and sequence type (ST) was determined via multilocus sequence typing (MLST). Results: UPEC strains (n = 126) from hospitalized children with complicated UTIs (cUTIs) were identified as O25b, of which 41.27% were multidrug resistant (MDR) and 15.87% were extensively drug resistant (XDR). The O25b strains harbored the fimH (95.23%), csgA (91.26%), papGII (80.95%), chuA (95.23%), iutD (88.09%), satA (84.92%), and intl1 (47.61%) genes. Moreover, 64.28% were producers of ESBLs and had high genetic diversity. ST131 (63.63%) was associated primarily with phylogenetic group B2, and ST69 (100%) was associated primarily with phylogenetic group D. Conclusion: UPEC O25b/ST131 harbors a wide genetic diversity of virulence and resistance genes, which contribute to cUTIs in pediatrics. Full article