Disc Diffusion Method Research Articles

Antibiotic profile classification of Proteus mirabilis using Machine Learning: An Investigation into Multidimensional Radiomics Features

Antimicrobial resistance (AMR) presents a significant threat to global healthcare. Proteus mirabilis causes catheter-associated urinary tract infections (CAUTIs) and exhibits increased antibiotic resistance. Traditional diagnostics still rely on culture-based approaches, which remain time-consuming. Here, we study the use of machine learning (ML) to classify bacterial resistance profiles using straightforward microscopic imaging of P. mirabilis for resistance classification integrated with radiomics feature analysis and ML models.From 150 P. mirabilis strains isolated from catheters of patients diagnosed with a CAUTI, 30 % displayed multidrug resistance using the standardized disk diffusion method, and 60% showed strong biofilm activity in microtiter plate assays. As a more rapid alternative, we introduce wavelet-based and regular microscopy imaging with feature extraction/selection, following image preprocessing steps (image denoising, normalization, and mask creation). These features enable training and testing different ML models with 5-fold cross-validation for P. mirabilis resistance classification. From these models, the Random Forest (RF) algorithm exhibited the highest performance with ACC=0.95, specificity=0.97, sensitivity=0.88, and AUC=0.98 among the other ML algorithms considered in this study for P. mirabilis resistance classification.This successful application of wavelet-based feature Radiomics analysis with RF model represents a crucial step towards a precise, rapid, and cost-effective method to distinguish antibiotic resistant P. mirabilis strains.

Evaluation and risk assessment of heavy metals in wetland soil in the University of Lagos

A study was conducted to evaluate the concentration of heavy metals, and the physicochemical and microbiological properties of wetland soils at the University of Lagos. Soil samples were randomly collected from the Faculty of Science (FSC), Lagoon Front, and Distance Learning Institute areas at depths of 5 cm, 10 cm, 15 cm, and 20 cm using a soil auger. The samples were stored in sterile nylon bags, appropriately labelled, and transported to the University of Lagos Central Research Laboratory for chemical analysis. The physicochemical characteristics measured included soil pH, determined using a pH meter, and moisture content, assessed through the oven-drying method. Heavy metal concentrations were analyzed using an Atomic Absorption Spectrophotometer. Microbial population diversity was examined using the disc diffusion method. Statistical analysis was performed using SPSS version 20, and mean values were evaluated using Tukey’s multiple comparison test at a probability level of P < 0.05. The results revealed that soil pH ranged from 7.20 to 8.81, with the highest value (8.81) recorded near the Faculty of Science (FSC-3). Eight heavy metals, including Pb, Ni, Cr, Cd, Fe, Cu, Mn, and Zn, were analyzed. Lead concentrations ranged from 0.02 to 0.29 ppm, followed by nickel, which ranged from 0.01 to 0.13 ppm. Chromium ranged from 0.0 to 0.25 ppm, cadmium from 0.03 to 0.08 ppm, iron from 1.02 to 2.96 ppm, copper from 3.31 to 4.60 ppm, manganese from 3.31 to 5.16 ppm, and zinc from 1.67 to 9.17 ppm. Lead and zinc concentrations were found to be below the permissible limit at all locations, while chromium and cadmium were within permissible limits. However, Zn, Mn, Cu, Fe, and Ni exceeded the permissible limits. Correlation analysis revealed a significant correlation (P < 0.05) between Pb and Zn, likely due to effluents from human waste and seepage from nearby dumpsites into the lagoon, streams, or beaches. The isolated microbial flora included three bacterial genera, Bacillus megaterium, Bacillus spp, and Enterobacter spp, as well as four fungal genera, Aspergillus, Penicillium, Aspergillus flavus, and Fusarium. Bacterial counts were lower than fungal counts in polluted soils, likely due to the nutrient status of the soil. The isolates were found to be tolerant to slightly alkaline pH and heavy metal pollutants, with microbial variation attributed to the impact of pH and heavy metals on the microbial population.

Antibiotic resistance and the search for resistance and virulence genes in Escherichia coli strains isolated from agropastoral farms (Poultry, Cattle, and Fish) In Southern Benin

Resistance of animal-origin enterobacteria to antibiotics is experiencing a concerning global increase, with an escalating impact of extended-spectrum beta-lactamases (ESBL). Finding solutions to this problem requires reliable and diverse data. It is in this context that the present study was initiated to combat infections caused by Escherichia coli in agropastoral farms (poultry, cattle, and fish) in southern Benin. Samples consisting of cattle dung, poultry droppings, as well as water and sludge from fish farms were aseptically collected for various laboratory analyses. A total of 714 E. coli strains were isolated from 1,376 collected samples, with 434 strains from the Atlantique department and 280 strains from the Zou department. The isolation of strains was carried out according to the NF V08-050: 2009 standard. The identified strains were tested against various antibiotics using the Bauer and Kirby disk diffusion method on agar, as recommended by the WHO. Strains resistant to one or more antibiotics were selected for the search for resistance and virulence genes. Furthermore, multidrug resistance of the strains to antibiotics was also observed, particularly in the Beta-lactam class (100% resistance to AMC) and tetracyclines (90% resistance to tetracyclines). Additionally, genotypic detection revealed a total absence of virulence genes, whereas resistance genes such as beta-lactamase Sulfhydryl Variable (bla-SHV), beta-lactamase Temoniera (bla-TEM), and quinolone resistance gene B (qnrB) were more frequently detected at respectively 58%, 82% and 60%.

A single-center analysis of clonal transmission of carbapenem-resistant Acinetobacter baumannii among intensive care unit patients during the COVID-19 pandemic

Carbapenem-Resistant Acinetobacter baumannii (CRAB) outbreak in intensive care units (ICUs) is a significant problem for healthcare facilities. In this study, we aimed to investigate the occurrence of CRAB isolates among ICU-admitted patients during the three waves of the COVID-19 pandemic in Iran using Multiple-Locus Variable Number Tandem-Repeat Analysis (MLVA). We obtained 50 (A) baumannii isolates from tracheal aspirate and blood culture samples. In the disc diffusion method, all isolates were cefotaxime, ceftriaxone, and cefepime-resistant, while 98% (49/50) of isolates were resistant to piperacillin, piperacillin-tazobactam, ceftazidime, and ciprofloxacin. Levofloxacin and tobramycin resistance was found in 76% (38/50) of isolates. In the microbroth dilution test all isolates were resistant to imipenem, 98% (49/50) to meropenem, 68% (34/50) to colistin, and 20% (10/50) to polymyxin (B) Based on the PCR findings, all isolates harbored blaOXA−40, ISAba-1, and int-2 genes. There were no isolates found that have the blaOXA−58, blaOXA−143, blaVEB−1, blaVIM, and int-3 genes. Among Extended-spectrum beta-lactamases (ESBL) genes, blaCTX−M, blaTEM, blaSHV, blaGES, and blaPER−1 have a prevalence of 42% (21/50), 84% (42/50), 58% (29/50), 78% (39/50), and 54% (27/50), respectively. 74% (37/50) of the isolates had the blaOXA−23 gene, while all of the isolates carried the blaOXA−40 gene. Among MBL genes, blaIPM, blaGIM, blaSIM, and blaNDM−1 have a prevalence of 20% (10/50), 8% (4/50), 22% (11/50), and 60% (30/50), respectively. The prevalence of int-1 was documented as 74% (37/50). Accordingly, all isolates were identified as CRAB. The co-existence of blaOXA−23/int-2 and blaOXA−23/isaba-1 was 74% (37/50). The co-existence of blaNDM−1/ISAba-1 was observed in 30 (60%) isolates. Using an 80% similarity threshold on the dendrogram constructed through MLVA typing, all isolates were grouped into two clusters: cluster A with 9 isolates from wave 3, and cluster B with 41 isolates from waves 3, 4, and 5. Our study confirms a clonal transmission of CRAB during the study period and suggests using molecular typing methods like MLVA in healthcare settings to identify dominant clones, antibiotic resistance patterns, and transmission routes. This will help to better manage the emergence and spread of antibiotic-resistant strains in future outbreaks.

Synthesis of Silver Nanoparticles with <i>Syzygium aromaticum</i> Leaves Extract as Antioxidant and Antimicrobial Materials

Silver nanoparticles (AgNPs) have attracted attention due to their unique properties and potential application. This research aimed to do green synthesis of AgNPs with Syzygium aromaticum leaves extract (SALE) and evaluate their antibacterial and antioxidant activities. Syzygium aromaticum leaves were extracted using distilled water at 70 °C for 30 min and the results were characterized with FTIR. AgNPs were synthesized by mixing AgNO3 precursor with SALE. The effects of parameters such as volume ratio of AgNO3 precursor to SALE, AgNO3 concentrations, and synthesis times were investigated. The synthesized AgNPs were characterized using UV-Vis spectrophotometer, FTIR, and TEM. Antibacterial activity of SALE and AgNPs was investigated against Escherichia coli (E.coli) and Bacillus subtilis (B. subtilis) with disc diffusion method and antioxidant activity was tested with DPPH method. The FTIR characterization revealed that SALE and resulting AgNPs contain O-H, C-H, C=O, C=C, C-O, and C≡C functional groups. The UV-Vis characterization demonstrated that AgNPs exhibited an absorption peak at λ = 420 nm indicating surface plasmon resonance. The optimal volume ratio of AgNO3 to SALE, AgNO3 concentrations, and synthesis time for AgNPs synthesis was achieved at 10:3, 5 mM, and 60 min respectively. TEM characterization indicated that AgNPs have spherical form and sizes ranging from 14 to 32 nm. The antibacterial testing revealed that AgNPs have antibacterial activities against E. coli and B. subtilis with inhibition zone values are 8,38 ± 0,48 and 6,88 ± 1,47 respectively. Additionally, antioxidant testing presented that the IC50 values were 85.05 µg/mL for SALE and 34.71 µg/mL for AgNPs. The results indicate that green synthesis of AgNPs from AgNO3 precursor with SALE was done successfully and this nanoparticle has good antibacterial and antioxidant activities.

Validation of EUCAST rapid antimicrobial susceptibility testing directly from positive blood cultures in a non-automated lab setting

ABSTRACT Introduction To speed up antimicrobial susceptibility testing (AST), the European Committee on Antimicrobial Susceptibility Testing (EUCAST) proposed rapid AST (RAST), a disk diffusion method to be read after 4, 6 and 8 hours of incubation. We investigated the feasibility of implementation of RAST in a non-automated lab setting. Materials & methods To this end, reference strains as well as a variety of clinical and resistant strains were used to spike sterile hemocultures (BioMérieux BACT/ALERT 3D® and Becton Dickinson BACTEC FX® systems), followed by RAST in comparison to classical long-incubation AST. Results & conclusion Our results with reference strains show that reading RAST after 4 hours is frequently too soon to obtain clinical results, and that Streptococcus pneumoniae reference strain did yield readable inhibition zones in RAST when harvested from BioMérieux BACT/ALERT 3D® bottle cultures. In a wider panel of strains, Gram positives RAST results were very similar to standard AST, while with Gram negative species errors were more frequently observed, limiting clinical implementation.

The battle against dental caries: defeating biofilm formed by bacterial isolates using vanillin and plant essential oils: in vitro and ex vivo approaches

BackgroundInfections caused by biofilm-forming bacteria have significantly linked to dental plaque and caries. The aim of this study is to assess efficacy of some natural compounds in inhibition and eradication of biofilm formed by bacterial isolates from dental infections.ResultsBacterial isolates were recovered from dental plaque/caries and identified using standard microbiological tests and 16S rDNA sequencing. The isolated bacterial strains include Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Klebsiella pneumoniae, and Escherichia coli. The antibiotic susceptibility was determined by disk diffusion method and revealed that the majority of isolates showed high antibiotic resistance, and 61% of isolates were found to be multidrug resistant. The biofilm formation capacity of isolates was investigated using microtiter plate assay. Among the 77 bacterial isolates, seventeen showed moderate biofilm formation capacity, twenty-two showed near-moderate, thirty-four had weak biofilm-forming capacity, and four were non-biofilm producers. The antibiofilm activity of tested compounds (rose and jasmine oils, propolis, vanillin, and vinegar) was evaluated against isolates with highest biofilm-forming capacity. The in vitro antibiofilm ability of tested substances were investigated alone or in combination with each other to evaluate their ability to prevent biofilm formation or destroy preformed single-/multispecies biofilms. Finally, antibiofilm ability of tested combination was evaluated ex vivo on natural teeth. Our results showed that vanillin in combination with rose or jasmine oils showed promising biofilm inhibition and biofilm eradication activities in both the in vitro and ex vivo models.ConclusionsDental plaque and caries can be successfully prevented using combination of vanillin with rose or jasmine oils, and these compounds can be incorporated in new anticaries dental formulations.

Isolation and Pattern of Antimicrobial Susceptibility Testing of Acinetobacter baumannii Clinical Isolates at Taksin Hospital: A Retrospective Study from 2019 to 2021

Objective: To determine the number of isolates and drug susceptibility testing of Acinetobacter baumannii at Taksin Hospital. Materials and Methods: A 3-year retrospective descriptive study was conducted between January 2019 and December 2021. Drug susceptibility testing of isolates to 13 drugs, which included amikacin, gentamycin, piperacillin/tazobactam, cefotaxime, ceftazidime, cefepime, three carbapenem drugs that were imipenem, meropenem, and doripenem, ciprofloxacin, trimethoprim-sulfamethoxazole, colistin, and tigecycline, were evaluated by standard disk diffusion method and automated microdilution method. Results: One thousand six hundred sixty-five non-duplicate A. baumannii isolates from different patients were detected and included 535 in 2019, 500 in 2020, and 630 in 2021. The patients’ ages ranged from three days to 100 years. The median age of patients was 70 years with an interquartile range of 56 to 80 years. A. baumannii was mostly found in patients aged older than 50 years old at 83.6%. They were isolated from sputum in 72.3%, urine in 9%, pus in 6.3%, tissue in 6.3%, and blood in 5.2%. Overall, A. baumannii demonstrated 91.5% and 79.8% susceptibility to colistin and tigecycline, respectively, but less susceptible to other drugs tested, with a range of 0.4% to 32.6%. Drug susceptibility to carbapenem such as imipenem, meropenem, and doripenem were 19.5%, 20%, and 20.9%, respectively. The carbapenem-resistant A. baumannii (CRAB) was found at 77.6%, 80%, and 79.1% in 2019, 2020, and 2021, respectively. For total isolates tested, the mean of CRAB was 78.9±1.2%. Conclusion: The present study results suggested the importance of monitoring A. baumannii at a hospital in Bangkok. Colistin and tigecycline had the high percentages of drug susceptibility rates in vitro. The antibiogram of susceptibility helps provide guidelines for clinician to consider empirical treatment.

Synthesis, Antimicrobial and Antioxidant Activity with in silico ADMET Prediction, Molecular Docking and Dynamics Studies of Carbazole Ring Containing Thiazole Schiff Bases

AbstractTwelve thiazole hybrids (2 a–2 l) were synthesized in this study incorporating with Schiff base and carbazole scaffolds and characterized by spectral analyses. Antimicrobial activity and antioxidant potency were investigated by agar disc diffusion method and DPPH free radical scavenging assays where trimethyldihydrothiazole 2 c had revealed its higher efficacy in both biological studies. It had displayed IC50 of 25.55±0.40 μg/mL which is higher than the standard ascorbic acid (IC50=41.31±1.83 μg/mL) as well as dihydrothiazoylchromenone 2 e showed promising antioxidant activity with IC50 value of 40.82±0.54. The compound 2 c satisfied all the ADME properties and toxicological parameters with maximum drug score value (0.39) among all derivatives. Molecular docking was undertaken for higher bioactive compounds where 2 c revealed its wide range of interactions with different amino acid residues which was also endorsed with the output of molecular dynamics simulation.

Comprehensive analysis of antioxidant and antibacterial activities of water and methanol extracts of Hibiscus flower

This study provides a comprehensive investigation into the antioxidant and antibacterial properties of Hibiscus flower extracts, highlighting their potential for future applications in functional foods. The research addressed key challenges by employing a multi-faceted approach, including four distinct methods for evaluating antioxidant potential: DPPH and FRAP assays, reducing power, and chelating ability of Fe ions. Total phenolic content was quantified using the Folin–Ciocalteu method, while tannins and flavonoids were assessed via vanillin–HCl and aluminum chloride assays. Additional analyses included spectrophotometric measurement of total flavonols and anthocyanins. Antibacterial activity was determined using a modified agar disk diffusion method, with results indicating significant inhibition of Salmonella typhimurium and Staphylococcus aureus by both aqueous and methanolic extracts. Colorimetric analysis revealed a reduction in chroma and hue angles, while FTIR spectroscopy identified key functional groups, such as suberin, lipids, and polysaccharides. Notably, the antioxidant efficacy varied depending on the solvent used, with aqueous extracts showing higher total phenolic content (4325.12 mg GAE/100 g) compared to methanolic extracts (3487.05 mg GAE/100 g). The study concludes that Hibiscus flower extracts, rich in antioxidants like tannins and anthocyanins, possess significant potential for use as natural preservatives and colorants in food products, supporting their role in developing novel functional foods.

Emergence of highly virulent and multidrug-resistant Escherichia coli in breeding sheep with pneumonia, Hainan Province, China

BackgroundSheep are a rarely raised livestock in Hainan Island, China, because of the unfavorable tropical marine climate. Here, this article reports a severe pneumonia in the sheep breeding and domestication facility caused acute mortality during the winter 2021–2022.MethodsSix sheep were clinically dissected and histopathologically observed. The bacteria were isolated and cultured by traditional methods and identified by 16S rRNA sequencing. The genotypes, serotypes, virulence genes and antimicrobial resistance genes were analyzed by PCR and whole genome sequencing. The pubMLST website was used for phylogenetic analysis of related strains. Kirby-Bauer disk diffusion method was used for antimicrobial susceptibility test. The antimicrobial susceptibility test standard was referred to the Clinical and Laboratory Standards Institute (CLSI). The virulence of bacteria was detected by mouse infection model.ResultsEtiology and histopathology examination of the pneumonia reveled pulmonary abscess and alveolar neutrophilia and pulmonary fibrinous exudates. Escherichia coli was the only bacterial species isolated, primarily from the lungs and blood of the six dead or moribund sheep, a total of 29 E. coli strains were isolated. Antimicrobial resistance profiling shows that all the isolates were resistant to six agents (penicillin, ampicillin, cephalothin, neomycin, erythromycin, and vancomycin) belonging to five classes of antibiotics, classifying them as multi drug resistant (MDR). Furthermore, genotyping analysis revealed all strains were common with 11–17 virulence factors indicating high pathogenicity. The lab mice infection model shows that all strains severely affect the health status particularly weight loss, lethargy, pneumonia and shortly lead to death. The molecular epidemiological analysis indicated most strains share the same genotype as previously reported strains in humans and other farmed animals this suggests a high possibility of cross-species transmission (CST) of virulent and MDR isolates. This CST could be from sheep to humans and other farmed animals or from humans and other farmed animals to sheep.ConclusionTherefore, this study indicates that E. coli is an emerging threat that causes sheep pneumonia in Hainan, and the quarantine of contacts is important to control the spread of virulent E. coli and the transmission of acquired resistance genes between humans and farmed animals such as sheep.

Antimicrobial Resistance and Pathotypes of Escherichia coli Isolates from Yellow-Legged Seagulls (Larus michahellis) in Central Italy

Seagulls are synanthropic wild birds that can contaminate, through their droppings, beaches, urban and peri-urban environments. This concern is more serious when seagulls eliminate antimicrobial-resistant pathogenic bacteria. This study analyzed the fecal samples from 137 yellow-legged seagulls (Larus michahellis) from Central Italy. A total of 218 Escherichia coli strains were isolated and analyzed for phenotypic and genotypic antimicrobial resistance and to identify the virulence genes characterizing different pathotypes. The disk diffusion method on all isolates found relevant resistance rates to ampicillin (38.99%), tetracycline (23.85%), and enrofloxacin (21.10%). On the basis of all results obtained with this test, 62 (28.44%) isolates were classified as multidrug-resistant (MDR) and 6 (2.75%) as extensive drug-resistant (XDR). Molecular analyses conducted on the strains phenotypically resistant to carbapenems, cephalosporins, and penicillins found 9/37 (24.32%) strains positive for blaOXA-48, 52/103 (50.49%) for blaTEM, 12/103 (11.65%) for blaCMY2, 3/103 (2.91%) for blaCTX, and 1/103 (0.97%,) for blaSHV. PCR to detect virulence genes characterizing different pathotypes found that 40 (18.35%) isolates had the astA gene, indicative of the enteroaggregative (EAEC) pathotype, 2 (0.92%) had cnf1, 2 (0.92%) had cnf2, and 1 (0.46%) had cdt-IV. All five (2.29%) strains were reportable as necrotoxigenic (NTEC), while 4 (1.83%) had both eaeA and escV, reportable as enteropathogenic (EPEC). Measures to limit seagulls’ access where humans and other animals reside are pivotal to reduce the risk of infection with antimicrobial-resistant and pathogenetic E. coli strains.

Increased incidence of methicillin resistant Staphylococcus aureus and methicillin resistant Staphylococcus epidermidis in the skin and nasal carriage among healthcare workers and inanimate hospital surfaces after the COVID-19 pandemic

Background and Objectives: Healthcare workers in hospitals are exposed to infectious diseases that occur in the hospital making them a source of infection for the patients. It is interfaced as cross-contamination agents for MRSA and MR-CoNS, and preventive measures need to be adapted accordingly. The study aimed to assess Methicillin-Resistant Staphylococcus (MRS) on the skin and nasal cavities of healthcare workers (HCWs) and identifying isolates to the species level. Materials and Methods: Swab samples were cultured on mannitol salt agar (MSA) to obtain MRS and determine their ability to produce coagulase. Their susceptibility to antibiotics were determined by agar screening and disk diffusion methods and further identification was done at the species level. Results: The highest percentage of methicillin resistant coagulase positive Staphylococci (MRCoPS) was reported among skins of male HCWs, (71.4%) were identified as MRSA. The highest levels of methicillin resistant coagulase negative Staph-ylococci (MRCoNS) were mainly detected in both nasal cavities, (75%) were identified as MRSE. MRSA was reported from doctors (p-value 0.033), whereas the highest incidence of MRSE was obtained from the nurses (p-value 0.048). Conclusion: This study highlighted that incidence of MRSA was mainly detected in doctors and MRCoNS in both nasal cav- ities. The highest percentage of MRCoNS was recovered from the patients’ room followed by the reception table. Moreover, vancomycin is suggested to be highly effective in managing and controlling S. aureus, MRSA- and MRSE related infections.

Synthesis and application of WO3/PVA Nano composite for Antibacterial and Thermal Insulation

Abstract A nanoparticle of tungsten trioxide (WO3) offers significant potential in enhancing both energy efficiency and human health, due to its natural antibacterial characteristics. The primary objective of this project revolves around the synthesis of Nano Tungsten trioxide particles via the chemical precipitation method. Developed WO3 NPs Characterized by using X-ray diffraction for; crystal structure, compound composition, and average crystalline size. Scanning Electron Microscopy (SEM) employed to analyze the shape and dimensions of the WO3 NPs. Explored the potential thermal insulation and antibacterial attributes by coating 1%, 3%, 5% and 7% concentration of synthesized WO3 NPs in polyvinyl alcohol onto a sodium silicate glass substrate (4x4 inches; 2 mm thickness) by using 50µm doctor blade. WO3/PVA solution showed the antibacterial properties measure by agar disc diffusion method on gram positive and negative bacterial culture S. coli and E. bacterial culture and the obtained results after incubation were showing very minor zone of incubation as compare to previous studies. Thermal insulation of coated glass samples were increased with % of WO3 NPs concentration in PVA and validate by significant coefficient of determination (R2=0.98).

Assessing the spread of sulfachloropyridazine in poultry environment and its impact on Escherichia coli resistance

Sulfachloropyridazine (SCP) is an antimicrobial (AM) commonly used in the poultry industry. This drug is excreted as the original compound, which may accumulate in litter. This work was done to assess whether SCP residues from droppings of broiler chickens that were treated with therapeutic doses of this drug spread into the production environment and to determine if these events were associated with the selection of resistant bacteria. To this end, broiler chickens were raised under controlled conditions, and their droppings and litter were processed to detect and identify SCP residues using an HPLC-MS/MS technique. This study selected Escherichia coli as an indicator bacterium for AM resistance. Its phenotypic resistance was determined using the Kirby-Bauer disk diffusion method, and its genotypic resistance was determined by performing a conventional PCR test. Our results showed that SCP residues did spread from the treated group to untreated sentinel groups because SCP residues in the litter reached levels up to 43.05 µg·kg−1 in a group placed immediately adjoining to the treated group, while another group placed 30 cm away showed a concentration of 29.79 µg·kg−1. Meanwhile, only trace concentrations were detected in droppings collected from sentinel groups. Of 239 strains of E. coli isolated from droppings, 12.13 % were resistant to sulfonamides, whereas 23.91 % of 92 E. coli isolated from broiler litter were resistant. The most prevalent resistance gene was the sul2 gene, both in droppings and litter, followed by the sul1 gene. The SCP concentrations were associated with the probability of E. coli being resistant to sulfonamides (p-value = 0.01). A Multiple Correspondence Analysis (MCA) also showed that phenotypic and genotypic resistances were associated and that both genes sul1 and sul2 would determine phenotypic resistance to sulfonamides in E. coli. The results presented in this study show that inedible by-products of the poultry industry are potentially a source of drug resistance that can spread from the animal production line to the environment, so awareness of the correct use of antimicrobials is essential to combat antimicrobial resistance.

The Comparison of Antibacterial Efficacy, Cell Proliferation, and Wound Healing Properties in Extracts Derived from Leaves and Inflorescences of Hang Kra Rog Phu Phan ST1 (Cannabis sativa L.)

The therapeutic potential of Cannabis sativa extracts in wound healing applications has garnered significant interest due to their diverse pharmacological properties. This study aimed to comparatively evaluate the antibacterial properties and wound-healing activity of ethanolic extracts derived from C. sativa leaves and inflorescences of the Hang Kra Rog Phu Phan ST1 cultivar. The extracts were prepared using ethanol, and their antibacterial properties, cytotoxicity and ability to promote cell proliferation and migration were investigated. The antibacterial activity was assessed against Staphylococcus epidermidis TISTR 518, Klebsiella pneumoniae TISTR 1383, Pseudomonas aeruginosa TISTR 2370, Staphylococcus aureus TISTR 1466 and Bacillus cereus TISTR 2373 using the disk diffusion method and broth microdilution assay. Cell viability and proliferation were evaluated using the MTT assay on human fibroblasts in serum-supplemented and serum-depleted media. The scratch wound healing assay was employed to quantify cell migration and wound closure. The results demonstrated that both leaf and inflorescence extracts exhibited antibacterial activity, with the inflorescence extract showing higher efficacy against S. aureus. Furthermore, the extracts enhanced fibroblast proliferation and migration in a concentration-dependent manner. Notably, the leaf extract outperformed the inflorescence extract in promoting fibroblast proliferation and migration, particularly at earlier time points. These findings suggest that C. sativa extracts, particularly from the leaves, possess wound-healing properties and could be developed as novel therapeutic agents for promoting efficient wound repair and managing chronic non-healing wounds. HIGHLIGHTS This research is the inaugural exploration into the antibacterial properties and wound healing capabilities of the Cannabis sativa L. Hang Kra Rog Phu Phan ST1 cultivar, a unique variant of Cannabis sativa. This strain has received official recognition and registration from the Rajamangala University of Technology Isan, Thailand. Our findings reveal that extracts derived from the leaves and inflorescences of the Hang Kra Rog Phu Phan ST1 strain display potent antibacterial activity against five tested pathogenic bacteria, namely Staphylococcus epidermidis TISTR 518, Klebsiella pneumoniae TISTR 1383, Pseudomonas aeruginosa TISTR 2370, Staphylococcus aureus TISTR 1466, and Bacillus cereus TISTR 2373. Moreover, these extracts stimulate the growth and movement of human fibroblasts. These compelling results underscore the potential for harnessing this plant as an efficacious agent for wound healing. GRAPHICAL ABSTRACT

The impact of the COVID-19 pandemic on pediatric bloodstream infections and alteration in antimicrobial resistance phenotypes in Gram-positive bacteria, 2020–2022

BackgroundAlteration in the etiology of pediatric bloodstream infections (BSIs) and antimicrobial resistance (AMR) is not well known during the Coronavirus disease 2019 (COVID-19) pandemic. This study aimed to investigate the impact of the COVID-19 pandemic on pediatric BSIs and alteration in antimicrobial resistance phenotypes in Gram-positive bacteria.MethodsThe frequency of BSIs among children under 18 years old was retrospectively recorded in a tertiary children’s hospital in Tehran, Iran from February 2020 to December 2022. The status of COVID-19 infection using reverse transcription polymerase chain reaction, bacteremia/fungemia according to BACTEC 9120 Culture System results, characterization of bacteria using biochemical tests, and antimicrobial susceptibility patterns for Gram-positive bacterial isolates using disk diffusion method were determined. Statistical analysis was done to measure the correlation of COVID-19 infection with BSIs and AMR.ResultsOut of 13,345 COVID-19 tests and 4,194 BACTEC blood culture requests, bacteremia/fungemia were confirmed in 10.37% (435/4,194) of the patients who requested both tests simultaneously. The COVID-19 infection was confirmed in 25.3% (110/435) of the patients with bacteremia/fungemia. The infection with characterized Gram-positive bacteria (GPB) and fungi was detected in 32.3% (140/433) and 8.31% (36/433) of the cases, respectively. Coagulase-negative Staphylococcus (CNS, 72, 16.62%), S. aureus (36, 8.3%), and Enterococcus spp. (22, 5%) were among the common isolates. Candida spp. and non-Candida yeasts were detected in 6.7% and 13.4% of the cases, respectively. A positive correlation was shown between the CNS bacteremia and COVID-19 infection (p-value = 0.019). Antibiotic susceptibility testing results showed the highest frequency of resistance to azithromycin among CNS, azithromycin and tetracycline among S. aureus and tetracycline among Enterococcus spp. Methicillin-resistance phenotype in the S. aureus (MRSA) and coagulase-negative Staphylococcus spp. (MR-CNS) was detected in 40% and 61.5% of the strains, respectively and the Enterococci were resistant to vancomycin in 33.3% of the isolates.ConclusionA decline in the trend of BSIs by GPB and an increase in AMR was shown in children during the COVID-19 pandemic. Increasing antibiotic resistance is a concern; however, chloramphenicol, linezolid, and vancomycin remain active against common causes of GPB-BSIs.

Investigating the impact of nanoplastics in altering the efficacy of clarithromycin antibiotics through In vitro studies

Plastics have significant global implications due to their environmental contamination from extensive use and improper disposal. Among plastic particles, nanoplastics (<1 μm) pose notable risks to organisms and ecosystems due to their high surface area, reactivity, and potential to carry environmental pollutants. This study explores the interaction between polystyrene nanoplastics (PSNPs) and clarithromycin (CLA), a broad-spectrum antibiotic, focusing on their combined impact on insulin (INS) and antibiotic-resistant (AMR) bacteria. PSNPs can adsorb CLA, leading to structural changes in insulin and affecting its physiological functions, potentially causing insulin resistance. Additionally, PSNPs reduce CLA's inhibitory effects on pathogenic bacteria, facilitating antibiotic resistance. Our research utilized UV–Vis Spectroscopy, FTIR, Fluorescence spectroscopy, and Circular dichroism (CD) spectroscopy to assess INS structural changes, alongside the Kirby-Bauer disk diffusion method for microbiological examination. The findings highlight the synergistic and antagonistic effects of PSNPs and CLA, underscoring the enhanced toxicity of CLA when adsorbed onto PSNPs and the complex interactions affecting both human health and bacterial resistance. Further studies are essential to fully understand these mechanisms and their broader implications.

Occurrence and characterization of ESKAPE organisms on the hands of veterinary students before patient contact at a veterinary academic hospital, South Africa

ObjectiveThis study aimed to investigate the presence of ESKAPE organisms on the hands of students working in the intensive care unit (ICU) at a veterinary academic hospital.MethodsA cross-sectional study was conducted among students working in an ICU at a veterinary academic hospital in South Africa. Students were sampled before the start of the ICU shift using a modified glove-juice method. Standard microbiological techniques and a series of polymerase chain reaction (PCR) assays were used to identify and characterize the bacteria. All the isolates were tested for resistance against a specific panel of antibiotics using the disk diffusion method. Proportions of bacterial species and their antimicrobial-susceptibility profiles were calculated.ResultsAt screening, all the veterinary students (n = 62) carried at least one of the ESKAPE organisms on their hands. Escherichia coli was the most isolated organism (76%, 47/62), followed by P. aeruginosa (48%, 30/62), A. baumannii (47%, 29/62), E. faecium (35%, 22/62), K. pneumoniae (27%, 17/62), and S. aureus (24%, 15/62). A reduced proportion of isolates were recovered from the samples, E. coli (26%, 12/47), E. faecium (23%, 5/22), P. aeruginosa (43%, 13/30), A. baumannii (24%,7/29), K. pneumoniae (41%, 7/17), and S. aureus (20%, 3/15). Most of the organisms showed a high proportion of resistance to at least one antibiotic. Multidrug resistance was reported among just over half (56%, 5/9) of E. coli, 40% (2/5) of E. faecium, 100% (13/13) of P. aeruginosa, and 33% (1/3) of S. aureus isolates.ConclusionStudents working in the ICU carry several organisms belonging to the ESKAPE group of organisms before contact with patients. Moreover, MDR resistance was common among this group of organisms. The findings of the present study underscore the importance of infection prevention and control (IPC) strategies to help reduce the likelihood of the spread of these organisms to personnel, owners, family members, and patients.

Identification and Assessment of Secondary Metabolites from Three Fungal Endophytes of Solanum mauritianum Against Public Health Pathogens

Fungal endophytes, symbiotic microorganisms residing within plants, are renowned for producing bioactive secondary metabolites with diverse beneficial properties. We investigated the antimicrobial potential of fungal endophytes isolated from Solanum mauritianum, an invasive weed, against clinically significant bacterial pathogens. Selected fungal endophytes (Penicillium chrysogenum, Fusarium sp., and Paracamarosporium leucadendri) were isolated from the plant’s leaves and fruits. Their crude extracts were tested against various referenced strains, such as Mycobacterium species (M. smegmatis ATCC 607 and M. bovis ATCC 27290), Staphylococcus aureus ATCC 6571, Bacillus subtilis ATCC 11774, Klebsiella species (K. pneumoniae ATCC 10031 and K. oxytoca ATCC 8724), Escherichia coli ATCC 10536, and Pseudomonas aeruginosa ATCC 10145, using the Kirby-Bauer disk diffusion method. Resazurin Microtiter Assay was used for the determination of the minimum inhibitory concentration. The chemical nature of the secondary metabolites in the crude extracts produced by fungal endophytes was evaluated using high-resolution liquid chromatography–mass spectrometry (LC-MS) using water and acetonitrile gradient. Liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS/MS) was employed for untargeted metabolomics. LC-QTOF-MS/MS identified 63 bioactive compounds across the three endophytes. P. chrysogenum had the highest activity against S. aureus and M. smegmatis (1.15 mg/mL and 0.02 mg/mL, respectively), while P. leucadendri demonstrated moderate activity against M. smegmatis (2.91 mg/mL) and E. coli (1.16 mg/mL). Fusarium sp. exhibited the broadest spectrum of antibacterial activity, with MIC values ranging from 0.03 mg/mL (B. subtilis) to 10 mg/mL (M. smegmatis). P. leucadendri produced 29 metabolites, Fusarium sp. had 23 identified metabolites, and a total of 11 metabolites were identified from P. chrysogenum. The fruits of the plant, accounting for 60%, appeared to be the most abundant in the endophyte diversity when compared to the stems and leaves. This study highlights the potential of fungal endophytes from S. mauritianum as a source of novel bioactive compounds, particularly against multidrug-resistant pathogens, contributing to the ongoing efforts to combat antimicrobial resistance.