Antibiotic-resistant Bacteria Research Articles

Bacteriological Evaluation of Surface and Groundwater used for Domestic Purposes in Ibadan

The right to safe and clean drinking water is a human right, unfortunately many communities rely on unsafe and contaminated water for drinking and domestic purposes. Through improper sewage and hospital waste disposal, pathogenic and antibiotic resistant bacteria have found their way to natural water sources used by humans. In this study we aim to profile pathogenic bacteria and resistant Escherichia coli (E. coli) from surface and ground water sources in three local government areas of Oyo state. 30 water samples were collected randomly from the selected local government areas. Bacteriological, biochemical and antibiotic-resistant analysis was carried out on the isolated bacteria from the water samples. E. coli (28.30%) and Kleb pneumoniae (23.77%) were the predominant bacteria isolates, while Shigella (16.22%) and Salmonella (16.22%) also had substantial percentages. Nineteen E. coli was isolated and identified, with high antibiotic resistance rates observed in nalidixic Acid (100%), augmentin (100%), ampiclox (94.74%) and third-generation cephalosporins. Conversely, moderate levels of resistance were observed in nitrofurantoin (55.55%), cefepime (57.89%), and second-generation fluoroquinolones. This study reveals substantial pathogenic bacteria and antibiotic resistant E. coli in water samples which endangers the health of communities’ dependent on these water sources and exposes the inhabitants to antibiotic-resistant organisms from contaminated water used for domestic purposes. It is crucial to put in place water treatment measures and quality monitoring programmes in order to guarantee that the populace has access to clean and safe drinking water.

Occurrence and Removal Efficiencies of Four Antibiotics in Kisii and Kabarnet Waste Water Treatment Plants, Kenya

Background: Antibiotics are presently considered as emerging contaminants with adverse effects in the environment and the population such as the development of antimicrobial resistant genes (ARG) and antimicrobial resistant bacteria (ARB). This study was guided by objective entailing to determine the seasonal occurrences and removal efficiencies of four antibiotics in Kisii and Kabarnet waste water treatment plants in Kenya.  
 Place and Duration of Study: Waste water samples were picked from Suneka wastewater treatment plant in Kisii county and Kabarnet Level V Hospital in Baringo County Kenya in the months of June and December 2020
 Methodology: Multiple samples were picked in Kisii and Kabarnet waste water treatment plants in the months of June and December correspond to the dry and wet seasons in Kenya. Collected waste water samples were centrifuged and filtered with glass microfiber filter papers and subsequently passed through a Solid phase extractor cartridge. High Performance Liquid Chromatography was used for quantification of antibiotics as per international commission for harmonization and subsequently applied for analysis.
 Results: The results revealed that the dry season had higher antibiotic concentrations at 1.29, 0.09, 2.92 and 1.82 µg/l for sulphamethoxazole, trimethoprim, ampicillin and amoxicillin respectively for the Kisii waste water treatment plant and 0.18, 0.05, 1.34 and 0.09 µg/l respectively for Kabarnet waste water treatment plant. During the wet seasons the measured concentrations were 1.11, 0.14, 2.04 and 1.34 µg/l for sulphamethoxazole, trimethoprim, ampicillin and amoxicillin at the Kisii WWTP, and 0.14, 0.06, 1.01 and 0.09 µg/l for Kabarnet WWTP. The removal efficiencies in the WWTPs, ranged from a high of 94 % to a low of 11.11 % depending on the type of antibiotic in both wet and dry seasons.
 Conclusion: Amoxicillin, ampicillin, trimethoprim and sulphamethaxazole were found in both Kisii WWTPs and Kabarnet WWTPs with nearly all antibiotics having a higher concentration of in the dry season than in the wet season. The highest calculated percentage removal was 94.03 % while the lowest calculated percentage removal was 7.14 %. The results suggest that the two WWTPs are effective for the removal of different types of antibiotics.

Effect of composting and storage on the microbiome and resistome of cattle manure from a commercial dairy farm in Poland.

Manure from food-producing animals, rich in antibiotic-resistant bacteria and antibiotic resistance genes (ARGs), poses significant environmental and healthcare risks. Despite global efforts, most manure is not adequately processed before use on fields, escalating the spread of antimicrobial resistance. This study examined how different cattle manure treatments, including composting and storage, affect its microbiome and resistome. The changes occurring in the microbiome and resistome of the treated manure samples were compared with those of raw samples by high-throughput qPCR for ARGs tracking and sequencing of the V3-V4 variable region of the 16S rRNA gene to indicate bacterial community composition. We identified 203 ARGs and mobile genetic elements (MGEs) in raw manure. Post-treatment reduced these to 76 in composted and 51 in stored samples. Notably, beta-lactam, cross-resistance to macrolides, lincosamides and streptogramin B (MLSB), and vancomycin resistance genes decreased, while genes linked to MGEs, integrons, and sulfonamide resistance increased after composting. Overall, total resistance gene abundance significantly dropped with both treatments. During composting, the relative abundance of genes was lower midway than at the end. Moreover, higher biodiversity was observed in samples after composting than storage. Our current research shows that both composting and storage effectively reduce ARGs in cattle manure. However, it is challenging to determine which method is superior, as different groups of resistance genes react differently to each treatment, even though a notable overall reduction in ARGs is observed.

Beef cattle feedlot runoff impacts on soil antimicrobial resistance

AbstractField application of beef cattle feedlot runoff may transport manure‐borne microbes and antibiotic resistant bacteria to agricultural soils eventually impacting deeper soils and groundwater. To evaluate this potential, total soil, antibiotic resistance (AR), and fecal indicator bacteria (Escherichia coli and Enterococcus) and the presence/abundance of AR genes were examined to a depth of 1.8 m in an agricultural field receiving long‐term application of feedlot runoff and compared to a nearby pasture receiving no runoff. While plate counts of total soil bacteria and cefotaxime‐resistant, erythromycin‐resistant, and tetracycline‐resistant bacteria decreased with depth on both fields (p < 0.001) by an average 2‐log10 colony forming unit g−1 to 1.8‐m depth, field differences were only observed with greater abundances of total soil and erythromycin‐resistant bacteria (p ≤ 0.026) in the runoff‐amended versus control field soils. Soil bacterial and fecal indicator bacterial isolates evaluated phenotypically for resistance to 12 antibiotics varied in range and sensitivity. Using a culture‐independent approach, erm(C) and tet(Q) were detected using polymerase chain reaction in 31% and 58% of runoff‐field samples throughout the soil profile. Detection of erm(C) and tet(Q) in the control field soil profile was less frequent (0% and 11%, respectively). Two other genes, erm(A) and tet(X) were not detected in any soil samples. Based upon these results, long‐term applications of beef cattle feedlot runoff may increase the total abundance of microorganisms in the surface and shallow soil, but the relative enrichment of AR was dependent upon the type of resistance evaluated and, more specifically, the genes targeted for analysis.

Phenotypic and Genotyping Identification of Extended Spectrum Beta-Lactamase (ESBL) Producing Enterobacteriaceae Obtained from Animal Fecal Samples within Owerri Metropolis

Background: One of the major means for the spread or distribution of antibiotic resistance is through animal, either by consumption or as companions. Most of the antibiotic resistant bacteria investigated have been reported to exhibit multidrug resistance which can either be acquired or natural. The present study was targeted at investigating the epidemiology of extended spectrum beta-lactamase-producing Enterobacteriaceae. Using stratified randomized sample design, three hundred (n = 300) fecal samples [of cattle, and goats (n = 150 for each)] were collected from various abattoir in Owerri metropolis. The isolation, purification, and antibiotic sensitivity were carried out using standard procedures. The resistant genes were investigated using forward and reverse primers specific for the following genes blaTEM, blaCTX-M, and blaSHV.
 Results: The result showed 97.67 % growth of Enterobacteriaceae. The highest Enterobacteriaceae isolated was in cattle fecal sample (61.00 %), while 2.33 % of the total samples did not show any growth for Enterobacteriaceae. Only three Enterobacteriaceae species were identified: Escherichia coli, Proteus vulgaris, and Klebsiella pneumonia. Escherichia coli was the most abundant Enterobacteriaceae (42.32 %) and P. vulgaris was the least abundant (23.89 %). The isolates showed multiple resistances to most of the antibiotics tested. All the isolates were susceptible to Imipenem (IPM). The molecular characterization showed the presence of the three genes (blaTEM, blaCTX-M, and blaSHV) investigated. The blaCTX gene-band was the most prevalent (72.22 %) while the blaSHV gene-band was the lowest prevalent resistant gene (61.11 %).
 Conclusion: It was concluded that the genes: blaTEM, blaCTX-M, and blaSHV were present in Enterobacteriaceae species (K. pneumoniae, P. vulgaris, and E. coli) isolated from abattoir in Owerri metropolis, Imo State.

CRISPR-Cas System: A New Dawn to Combat Antibiotic Resistance.

Antimicrobial resistance (AMR) can potentially harm global public health. Horizontal gene transfer (HGT), which speeds up the emergence of AMR and increases the burden of drug resistance in mobile genetic elements (MGEs), is the primary method by which AMR genes are transferred across bacterial pathogens. New approaches are urgently needed to halt the spread of bacterial diseases and antibiotic resistance. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR), an RNA-guided adaptive immune system, protects prokaryotes from foreign DNA like plasmids and phages. This approach may be essential in limiting horizontal gene transfer and halting the spread of antibiotic resistance. The CRISPR-Cas system has been crucial in identifying and understanding resistance mechanisms and developing novel therapeutic approaches. This review article investigates the CRISPR-Cas system's potential as a tool to combat bacterial AMR. Antibiotic-resistant bacteria can be targeted and eliminated by the CRISPR-Cas system. It has been proven to be an efficient method for removing carbapenem-resistant plasmids and regaining antibiotic susceptibility. The CRISPR-Cas system has enormous potential as a weapon against bacterial AMR. It precisely targets and eliminates antibiotic-resistant bacteria, facilitates resistance mechanism identification, and offers new possibilities in diagnostics and therapeutics.

Rapid eradication of vancomycin and methicillin-resistant Staphylococcus aureus by MDP1 antimicrobial peptide coated on photocrosslinkable chitosan hydrogel: in vitro antibacterial and in silico molecular docking studies

IntroductionAntibiotic resistance and weak bioavailability of antibiotics in the skin due to systemic administration leads to failure in eradication of vancomycin- and methicillin-resistant Staphylococcus aureus (VRSA and MRSA)-associated wound infections and subsequent septicemia and even death. Accordingly, this study aimed at designing a photocrosslinkable methacrylated chitosan (MECs) hydrogel coated by melittin-derived peptide 1 (MDP1) that integrated the antibacterial activity with the promising skin regenerative capacity of the hydrogel to eradicate bacteria by burst release strategy.MethodsThe MECs was coated with MDP1 (MECs-MDP1), characterized, and the hydrogel-peptide interaction was evaluated by molecular docking. Antibacterial activities of MECs-MDP1 were evaluated against VRSA and MRSA bacteria and compared to MECs-vancomycin (MECs-vanco). Antibiofilm activity of MECs-MDP1 was studied by our novel ‘in situ biofilm inhibition zone (IBIZ)’ assay, and SEM. Biocompatibility with human dermal fibroblast cells (HDFs) was also evaluated.Results and DiscussionMolecular docking showed hydrogen bonds as the most interactions between MDP1 and MECs at a reasonable affinity. MECs-MDP1 eradicated the bacteria rapidly by burst release strategy whereas MECs-vanco failed to eradicate them at the same time intervals. Antibiofilm activity of MECs-MDP1 were also proved successfully. As a novel report, molecular docking analysis has demonstrated that MDP1 covers the structure of MECs and also binds to lysozyme with a reasonable affinity, which may explain the inhibition of lysozyme. MECs-MDP1 was also biocompatible with human dermal fibroblast skin cells, which indicates its safe future application. The antibacterial properties of a photocrosslinkable methacrylated chitosan-based hydrogel coated with MDP1 antimicrobial peptide were successfully proved against the most challenging antibiotic-resistant bacteria causing nosocomial wound infections; VRSA and MRSA. Molecular docking analysis revealed that MDP1 interacts with MECs mainly through hydrogen bonds with reasonable binding affinity. MECs-MDP1 hydrogels eradicated the planktonic state of bacteria by burst release of MDP1 in just a few hours whereas MECs-vanco failed to eradicate them. inhibition zone assay showed the anti-biofilm activity of the MECs-MDP1 hydrogel too. These findings emphasize that MECs-MDP1 hydrogel would be suggested as a biocompatible wound-dressing candidate with considerable and rapid antibacterial activities to prevent/eradicate VRSA/MRSA bacterial wound infections.

A prognostic insight of the mRNA vaccine against antibiotic-resistant bacteria

A prognostic insight of the mRNA vaccine against antibiotic-resistant bacteria

Use of Selected Plant Extracts in Controlling and Neutralizing Toxins and Sporozoites Associated with Necrotic Enteritis and Coccidiosis

Due to increasing concerns about the contamination of animal food products with antibiotic-resistant bacteria and their byproducts, phytogenic feed additives in animal diets have been explored as antibiotic alternatives. In this study, we investigated the effect of ginger root extract (GRE), green tea extract (GTEC caffeinated and GTED decaffeinated), and onion peel combined (OPEC) on the activity of C. perfringens toxin genes and Eimeria tenella sporozoites. To this end, two Clostridium perfringens strains, CP19 and CP240 (Rollins Diagnostic Lab, Raleigh, NC, USA), were cultured (three replicates per treatment) as follows: without additives (Control), with Bacitracin Methylene Disalicylate (BMD), with GRE, with GTEC, with GTED, and, finally, with OPEC for 0, 2, 4, 6, 8, and 24 h. RNA was extracted to determine the expression of tpeL, alpha toxin (α-toxin), and NetB and we measured the protein concentration of NetB-positive C. perfringens toxin. Also, we evaluated the cytotoxic effect of green tea and ginger extracts on E. tenella sporozoites. Results show that phytogenic extracts, GRE, GTEC, and GTED, significantly reduced (p < 0.05) the level of expression of α-toxin gene compared to control; however, BMD treatment showed much less effect. Furthermore, NetB and tpeL encoding gene expression was significantly (p < 0.05) reduced by GRE and GTED, as well as BMD treatment, compared to the control. In contrast, GTEC treatment did not change the expression levels of these genes and was similar to control. With the CP240 strain, all the selected phytogenic extracts significantly reduced (p < 0.05) the expression of selected genes, except for OPEC, which was similar to control. GRE, GTEC, and GTED all reduced the viability of concentration of E. tenella sporozoites. Overall, our data show that these selected phytogenic extracts reduced the level of expression of toxin encoding genes associated with necrotic enteritis and decreased the viability of sporozoites which cause coccidiosis in broiler chicken.

Addressing Challenges in Wildlife Rehabilitation: Antimicrobial-Resistant Bacteria from Wounds and Fractures in Wild Birds

Injuries and bone fractures are the most frequent causes of admission at wildlife rescue centers. Wild birds are more susceptible to open fractures due to their anatomical structure, which can lead to osteomyelitis and necrosis. Antibiotic therapy in these cases is indispensable, but the increase of antimicrobial-resistant isolates in wildlife has become a significant concern in recent years. In this context, the likelihood of antibiotic failure and death of animals with infectious issues is high. This study aimed to isolate, identify, and assess the antimicrobial resistance pattern of bacteria in wounds and open fractures in wild birds. To this end, injured birds admitted to a wildlife rescue center were sampled, and bacterial isolation and identification were performed. Then, antimicrobial susceptibility testing was assessed according to the disk diffusion method. In total, 36 isolates were obtained from 26 different birds. The genera detected were Staphylococcus spp. (63.8%), Escherichia (13.9%), Bacillus (11.1%), Streptococcus (8.3%), and Micrococcus (2.8%). Among Staphylococcus isolates, S. lentus and S. aureus were the most frequent species. Antimicrobial resistance was detected in 82.6% of the isolates, among which clindamycin resistance stood out, and 31.6% of resistant isolates were considered multidrug-resistant. Results from this study highlight the escalating scope of antimicrobial resistance in wildlife. This level of resistance poses a dual concern for wildlife: firstly, the risk of therapeutic failure in species of significant environmental value, and, secondly, the circulation of resistant bacteria in ecosystems.

Dissemination of antibiotic resistance genes from aboveground sources to groundwater in livestock farms

Antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) are prevalent in various environments on livestock farms, including livestock waste, soil, and groundwater. Contamination of groundwater by ARB and ARGs in livestock farms is a growing concern as it may have potentially huge risks to human health. However, the source of groundwater-borne ARB and ARGs in animal farms remains largely unknown. In this study, different types of samples including groundwater and its potential contamination sources from aboveground (pig feces, wastewater, and soil) from both working and abandoned swine feedlots in southern China were collected and subjected to metagenomic sequencing and ARB isolation. The source tracking based on metagenomic analysis revealed that 56–95 % of ARGs in groundwater was attributable to aboveground sources. Using metagenomic assembly, we found that 45 ARGs predominantly conferring resistance to aminoglycosides, sulfonamides, and tetracyclines could be transferred from the aboveground sources to groundwater, mostly through plasmid-mediated horizontal gene transfer. Furthermore, the full-length nucleotide sequences of sul1, tetA, and TEM-1 detected in ARB isolates exhibited the close evolutionary relationships between aboveground sources and groundwater. Some isolated strains of antibiotic-resistant Pseudomonas spp. from aboveground sources and groundwater had the high similarity (average nucleotide identity > 99 %). Notably, the groundwater-borne ARGs were identified as mainly carried by bacterial pathogens, potentially posing risks to human and animal health. Overall, this study underscores the dissemination of ARGs from aboveground sources to groundwater in animal farms and associated risks.

A Multidrug-Resistant Extended-Spectrum Beta-Lactamase (ESBL)-Producing Enterobacter hormaechei Strain from Mixed Sprouts.

Fresh vegetables can harbor antibiotic-resistant bacteria, including extended-spectrum β-lactamase (ESBL)-producing Enterobacterales. Enterobacter hormaechei is a bacterium belonging to the Enterobacterales order and the most commonly identified nosocomial pathogen of Enterobacter cloacae complex. The purpose of this study was to characterize a multi-drug resistant ESBL-producing E. hormaechei strain isolated from a sample of mixed sprouts. Vegetable samples were pre-enriched in buffered peptone water, followed by enrichment in Enterobacteria Enrichment Broth, and isolation on Chromagar™ ESBL plates. One isolate from a sprout sample was confirmed to produce both ESBL and AmpC β-lactamases through the combination disk diffusion assay using antibiotic disks containing cefotaxime and ceftazidime with or without clavulanate, and with or without cloxacillin, respectively. The isolate was also resistant to multiple antibiotics, including cefotaxime, ceftazidime, chloramphenicol, trimethoprim-sulfamethoxazole, tetracycline, gentamicin, ampicillin, and amoxicillin-clavulanate, as determined by antimicrobial susceptibility testing. Through whole genome sequencing, the isolate was identified as E. hormaechei 057-E1, which carried multiple antibiotic resistance (AR) genes and a sul2-aph(3″)-Ib-aph(6)-Id-blaTEM-1-ISEcp1 -blaCTX-M-15 gene cluster. Our results further demonstrate the important role of fresh vegetables in AR and highlight the need to develop strategies for AR mitigation in fresh vegetables.

Research Progress on the Combination of Quorum-Sensing Inhibitors and Antibiotics against Bacterial Resistance.

Bacterial virulence factors and biofilm development can be controlled by the quorum-sensing (QS) system, which is also intimately linked to antibiotic resistance in bacteria. In previous studies, many researchers found that quorum-sensing inhibitors (QSIs) can affect the development of bacterial biofilms and prevent the synthesis of many virulence factors. However, QSIs alone have a limited ability to suppress bacteria. Fortunately, when QSIs are combined with antibiotics, they have a better therapeutic effect, and it has even been demonstrated that the two together have a synergistic antibacterial effect, which not only ensures bactericidal efficiency but also avoids the resistance caused by excessive use of antibiotics. In addition, some progress has been made through in vivo studies on the combination of QSIs and antibiotics. This article mainly expounds on the specific effect of QSIs combined with antibiotics on bacteria and the combined antibacterial mechanism of some QSIs and antibiotics. These studies will provide new strategies and means for the clinical treatment of bacterial infections in the future.

Emerging Issues on Antibiotic-Resistant Bacteria Colonizing Plastic Waste in Aquatic Ecosystems

Antibiotic-resistant bacteria (ARB) adhesion onto plastic substrates is a potential threat to environmental and human health. This current research investigates the prevalence of two relevant human pathogens, Staphylococcus spp. and Klebsiella spp., and their sophisticated equipment of antibiotic-resistant genes (ARGs), retrieved from plastic substrates submerged into an inland water body. The results of microbiological analysis on selective and chromogenic media revealed the presence of colonies with distinctive phenotypes, which were identified using biochemical and molecular methods. 16S rDNA sequencing and BLAST analysis confirmed the presence of Klebsiella spp., while in the case of Staphylococcus spp., 63.6% of strains were found to be members of Lysinibacillus spp., and the remaining 36.3% were identified as Exiguobacterium acetylicum. The Kirby–Bauer disc diffusion assay was performed to test the susceptibility of the isolates to nine commercially available antibiotics, while the genotypic resistant profile was determined for two genes of class 1 integrons and eighteen ARGs belonging to different classes of antibiotics. All isolated bacteria displayed a high prevalence of resistance against all tested antibiotics. These findings provide insights into the emerging risks linked to colonization by potential human opportunistic pathogens on plastic waste commonly found in aquatic ecosystems.

Antibacterial therapy for kidney pathology of small pets

The article considers etiological factors, clinical manifestations, laboratory and instrumental methods for diagnosing kidney pathology, which requires the use of antibacterial drugs – pyelonephritis. However, this pathology can be difficult to diagnose and not every antibacterial drug is suitable for its treatment. The main ways of kidney damage by bacterial infection in small domestic animals are ascending infection and hematogenous introduction, when pathogenic microorganisms enter the kidneys along with the bloodstream. The article provides predisposing factors leading to the development of pyelonephritis. Pyelonephritis has several types of course. The acute form of the disease is most easily diagnosed due to the vivid clinical signs and pronounced changes in blood, urine, and kidney sonography tests. Whereas the chronic form and latent course may be overlooked due to the nonspecific clinical picture. In the treatment of pyelonephritis, the occurrence of antibiotic resistance in bacteria can be fatal for the patient, because if it is impossible to eliminate infection in the kidneys, the development of renal failure is inevitable, leading to all manifestations of uremia, anemia and other changes that occur with impaired renal function. To prevent the development of antibiotic resistance, it is extremely important to make a final diagnosis through bacteriological urine culture, but pyelonephritis is a pathology that requires a comprehensive diagnostic approach and, if suspected, it is advisable to start etiological treatment immediately, correctly choosing an antibiotic. Not every antibacterial drug is suitable for the treatment of pyelonephritis. For example, one of the first-line drugs of choice in the treatment of urinary tract infection (UTI), amoxicillin with clavulanic acid does not penetrate the renal parenchyma. The group of choice for the treatment of bacterial inflammation of the kidneys are fluoroquinolones, but not every drug from this group is suitable for treatment in animals and there are certain restrictions in their use in the case of puppies, kittens, as well as in cats as a species, therefore, the selection of an antibiotic should be carried out individually in each clinical case, taking into account the pharmacokinetics and pharmacodynamics of the drug, the result of urine sampling and other studies, as well as the physiological characteristics of the patient.

Etiology, pathogenesis, diagnosis and treatment of encrusted cystitis and pyelitis: Review of the problem from a historical perspective

Introduction: Encrusted cystitis and pyelitis share many similarities regarding etiology, pathogenesis, diagnosis and treatment. These processes usually occur on previously injured urological tissue with the participation of an infection by urease-producing bacteria. At present Corynebacterium urealyticum, a highly antibiotic-resistant bacteria, appears to be the pathogen most often involved in these processes. In this review we analyze the history of these diseases with special emphasis on the scientific advances of the last century. We also review the role of urease and bacterial infection as well as a brief update on the epidemiological, histopathological, diagnostic and therapeutic aspects. Relevance: The importance and severity of these pathologies that are frequently underdiagnosed and difficult to treat are stressed. Conclusions: The clinical history, exploratory and analytical data, which highlight alkaline urine, sometimes with an ammoniacal odor, pyuria, hematuria and the presence of magnesium ammonium phosphate crystals, should alert the urologist to the existence of an encrusting uropathy. Simple radiology, ultrasound, computed tomography and endoscopic visualization of the lesions confirm the diagnosis. Treatment generally requires surgery, use of antibiotics, urinary acidifiers and bacterial urease inhibitors. Keywords: cystitis, pyelitis, encrustations, infection

FORMATION OF NANOCONES AND GENERATION OF NEGATIVE POTENTIAL ON STAINLESS STEEL SURFACES BY ELECTROCHEMICAL ETCHING SYNERGISTICALLY REDUCE PSEUDOMONAS AERUGINOSA'S BIOFILM

Hospital-acquired infections cause severe patient problems because of the augmented appearance of antibiotic-resistant bacteria, including Pseudomonas aeruginosa. Material surfaces modified with several biophysical parameters can decrease bacterial biofilm formation, which could be an advantageous alternative to treatment with antibiotics. Since stainless steel is an extensively used material for manufacturing medical implants and in healthcare settings, in this study, we used stainless steel (SS 316L and SS 304) to examine the result of the material surface topographies on bacterial biofilm establishment. This work used the electrochemical etching method to modify the stainless steel surface topography as an anode. The electrochemical etching method influenced the nanocones’ formation on stainless steel surfaces of both SS316L (Disk-6: 2682 peaks/[Formula: see text]m[Formula: see text] and SS304 (Disk-12: 1654 peaks/[Formula: see text]m[Formula: see text] estimated by atomic force microscopy and 3D Profilometer reduced the biofilm by 78% and 85%, respectively. Additionally, the higher negative potential on an average of 600[Formula: see text]mV measured by Kelvin probe atomic force microscopy reduced the biofilm formation on both SS316L and SS304 surface synergistically compared to the non-electrochemically etched surface. Biofilm growth and nanopotrusions on the stainless-steel surface examined by atomic force microscopy and scanning electron microscopy demonstrated significantly dead bacterial cells (20%) on the electrochemically etched surface than on the non-electrochemically etched surface after 2[Formula: see text]h contact time. Our observations exhibit that the nanotextured surface topographies and surface negative potential effectively inhibit bacterial adhesion and biofilm formation.

Occurrence of Antimicrobial-Resistant Bacteria in Intestinal Contents of Wild Marine Fish in Chile

Antimicrobial-resistant bacteria (ARB) from the intestinal contents of wild fish may have a relevant ecological significance and could be used as indicators of antimicrobial-resistance dissemination in natural bacterial populations in water bodies impacted by urban contamination. Thus, the occurrence of ARB in the intestinal contents of pelagic and demersal wild fishes captured in anthropogenic-impacted Coquimbo Bay in Chile was studied. Culturable counts of total and antimicrobial-resistant bacteria were determined by a spread plate method using Trypticase soy agar and R2A media, both alone and supplemented with the antimicrobials amoxicillin, streptomycin, florfenicol, oxytetracycline and ciprofloxacin, respectively. Heterotrophic plate counts of pelagic and demersal fishes ranged from 1.72 × 106 CFU g−1 to 3.62 × 109 CFU g−1, showing variable proportions of antimicrobial resistance. Representative antimicrobial-resistant isolates were identified by 16S rRNA gene sequencing, and isolates (74) from pelagic fishes mainly belonged to Pseudomonas (50.0%) and Shewanella (17.6%) genera, whereas isolates (68) from demersal fishes mainly belonged to Vibrio (33.8%) and Pseudomonas (26.5%) genera. Antimicrobial-resistant isolates were tested for susceptibility to 12 antimicrobials by an agar disk diffusion method, showing highest resistance to streptomycin (85.2%) and amoxicillin (64.8%), and lowest resistance to oxytetracycline (23.2%) and ciprofloxacin (0.7%). Only furazolidone and trimethoprim/sulfamethoxazole were statistically different (p < 0.05) in comparisons between isolates from pelagic and demersal wild fishes. Furthermore, an important number of these isolates carried plasmids (53.5%) and produced Extended-Spectrum-β-lactamases (ESBL) (16.9%), whereas the detection of Metallo–β–Lactamases and class 1-integron was rare. This study provides evidence that wild fish are important reservoirs and spreading-vehicles of ARB, carrying plasmids and producing ESBLs in Chilean marine environments.

A metagenomic study of antibiotic resistance genes in a hypereutrophic subtropical lake contaminated by anthropogenic sources

Antibiotic resistance genes (ARGs) are a major threat to human and environmental health. This study investigated the occurrence and distribution of ARGs in Lake Cajititlán, a hypereutrophic subtropical lake in Mexico contaminated by anthropogenic sources (urban wastewater and runoff from crop and livestock production). ARGs (a total of 475 genes) were detected in 22 bacterial genera, with Pseudomonas (144 genes), Stenotrophomonas (88 genes), Mycobacterium (54 genes), and Rhodococcus (27 genes) displaying the highest frequencies of ARGs. Among these, Pseudomonas aeruginosa and Stenotrophomonas maltophilia showed the highest number of ARGs. The results revealed a diverse array of ARGs, including resistance to macrolides (11.55 %), aminoglycosides (8.22 %), glycopeptides (6.22 %), tetracyclines (4 %), sulfonamides (4 %), carbapenems (1.11 %), phenicols (0.88 %), fluoroquinolones (0.44 %), and lincosamides (0.22 %). The most frequently observed ARGs were associated with multidrug resistance (63.33 %), with MexF (42 genes), MexW (36 genes), smeD (31 genes), mtrA (25 genes), and KHM-1 (22 genes) being the most common. Lake Cajititlán is a recreational area for swimming, fishing, and boating, while also supporting irrigation for agriculture and potentially acting as a drinking water source for some communities. This raises concerns about the potential for exposure to antibiotic-resistant bacteria through these activities. The presence of ARGs in Lake Cajititlán poses a significant threat to both human and environmental health. Developing strategies to mitigate the risks of antibiotic resistance, including improving wastewater treatment, and promoting strategic antibiotic use and disposal, is crucial. This study represents a significant advancement in the understanding of antibiotic resistance dynamics in a hypereutrophic subtropical lake in a developing country, providing valuable insights for the scientific community and policymakers.

Development of a High-Throughput Analytical Method for Antimicrobials in Wastewater Using an Automated Pipetting and Solid-Phase Extraction System

Antimicrobial resistance (AMR) has emerged and spread globally. Recent studies have also reported the presence of antimicrobials in a wide variety of aquatic environments. Conducting a nationwide monitoring survey of AMR in the environment to elucidate its status and to assess its impact on ecosystems and human health is of social importance. In this study, we developed a novel high-throughput analysis (HTA) system based on a 96-well plate solid-phase extraction (SPE), using automated pipetting and an SPE pre-treatment system. The effectiveness of the system as an HTA for antimicrobials in environmental water was verified by comparing it with a conventional manual analytical system in a domestic hospital over a period of two years and four months. The results of the manual analysis and HTA using a combination of automated pipetting and SPE systems were generally consistent, and no statistically significant difference was observed (p > 0.05) between the two systems. The agreement ratios between the measured concentrations based on the conventional and HTA methods were positively correlated with a correlation coefficient of r = 0.99. These results indicate that HTA, which combines automated pipetting and an SPE pre-treatment system for rapid, high-volume analysis, can be used as an effective approach for understanding the environmental contamination of antimicrobials at multiple sites. To the best of our knowledge, this is the first report to present the accuracy and agreement between concentrations based on a manual analysis and those measured using HTA in hospital wastewater. These findings contribute to a comprehensive understanding of antimicrobials in aquatic environments and assess the ecological and human health risks associated with antimicrobials and antimicrobial-resistant bacteria to maintain the safety of aquatic environments.