Development and antimicrobial activity of composite edible films of chitosan and nisin incorporated with perilla essential oil-glycerol monolaurate emulsions.

Aquatic products are highly susceptible to spoilage, and preparing composite edible film with essential oil is an effective solution. In this study, composite edible films were prepared using perilla essential oil (PEO)-glycerol monolaurate emulsions incorporated with chitosan and nisin, and the film formulation was optimized by response surface methodology. These films were applied to ready-to-eat fish balls and evaluated over a period of 12 days. The films with the highest inhibition rate against Staphylococcus aureus were acquired using a polymer composition of 6 µL/mL PEO, 18.4 µg/mL glycerol monolaurate, 14.2 mg/mL chitosan, and 11.0 µg/mL nisin. The fish balls coated with the optimal edible film showed minimal changes in appearance during storage and significantly reduced total bacterial counts and total volatile basic nitrogen compared to the control groups. This work indicated that the composite edible films containing essential oils possess ideal properties as antimicrobial packaging materials for aquatic foods.

Integrating MALDI-TOF Mass Spectrometry with Machine Learning Techniques for Rapid Antimicrobial Resistance Screening of Foodborne Bacterial Pathogens.

Although MALDI-TOF mass spectrometry (MS) is considered as the gold standard for rapid and cost-effective identification of microorganisms in routine laboratory practices, its capability for antimicrobial resistance (AMR) detection has received limited focus. Nevertheless, recent studies explored the predictive performance of MALDI-TOF MS for detecting AMR in clinical pathogens when machine learning techniques are applied. This chapter describes a routine MALDI-TOF MS workflow for the rapid screening of AMR in foodborne pathogens, with Campylobacter spp. as a study model.

Genomic Pipeline for Analysis of Mutational Events in Bacteria.

Unveiling the strategies of bacterial adaptation to stress constitute a challenging area of research. The understanding of mechanisms governing emergence of resistance to antimicrobials is of particular importance regarding the increasing threat of antibiotic resistance on public health worldwide. In the last decades, the fast democratization of sequencing technologies along with the development of dedicated bioinformatical tools to process data offered new opportunities to characterize genomic variations underlying bacterial adaptation. Thereby, research teams have now the possibility to dive deeper in the deciphering of bacterial adaptive mechanisms through the identification of specific genetic targets mediating survival to stress. In this chapter, we proposed a step-by-step bioinformatical pipeline enabling the identification of mutational events underlying biocidal stress adaptation associated with antimicrobial resistance development using Escherichia marmotae as an illustrative model.

Bioinformatic Pipeline for Profiling Foodborne Bacterial Ecology and Resistome from Short-Read Metagenomics.

Next-generation sequencing revolutionized food safety management these last years providing access to a huge quantity of valuable data to identify, characterize, and monitor bacterial pathogens on the food chain. Shotgun metagenomics emerged as a particularly promising approach as it enables in-depth taxonomic profiling and functional investigation of food microbial communities. In this chapter, we provide a comprehensive step-by-step bioinformatical workflow to characterize bacterial ecology and resistome composition from metagenomic short-reads obtained by shotgun sequencing.

Epinecidin-1 and lactic acid synergistically inhibit Aeromonas hydrophila through membrane disruption.

Epinecidin-1 (Epi-1) is an antimicrobial peptide originated from fish with various pharmacological activities but carries the risk of acquiring resistance with long-term use. In the present study, we use L-lactic acid to enhance the antibacterial activity of synthesized Epi-1 against the aquaculture and food pathogen Aeromonas hydrophila. The results showed that 5.5 mmol/L lactic acid increased the inhibitory and bactericidal activity of 25 µmol/L Epi-1 against two strains of A. hydrophila. The laser confocal images proved that lactic acid pre-treatment improved the attachment efficiency of Epi-1 in A.hydrophila cells. In addition, lactic acid enhanced the damaging effect of Epi-1 on the cell membrane of A. hydrophila, evidenced by releasing more nucleic acids, proteins, and transmembrane pH ingredients decrease and electromotive force dissipation. SEM images showed that compared with the single Epi-1 treatment, the co-treatment of Epi-1 and lactic acid caused more outer membrane vesicles (OMVs) and more severe cell deformation. These findings proved that lactic acid could enhance the efficiency of Epi-1 against A. hydrophila and shed light on new aspects to avoid resistance of pathogens against Epi-1.

A Novel Bioprotective Strain of Lactiplantibacillus plantarum F2-Physiological, Genetical, and Antimicrobial Characterization.

Lactiplantibacillus plantarum is a member of lactic acid bacteria that improves the quality of fermented foods while also having a positive impact on human health. In this study, L. plantarum F2 was studied for characteristics such as biochemical and genetic identification, metabolite production, antimicrobial activity, and plasmid content. This strain exerts antimicrobial activity against some Gram-positive and Gram-negative pathogens (Listeria monocytogenes, Staphylococcus aureus, Salmonella, and Escherichia coli) with inhibition zone diameters ranging between 17.0 and 29.0 mm; it can ferment glucose, arabinose, galactose, lactose, and demonstrated the ability to grow at high temperature (50°C). Another physiological specification of the strain was the morphology of the isolate in selective medium, the de Man, Rogosa, Sharpe medium (MRS medium containing triphenyl tetrazolium chloride), which exhibits a chromogenic colony (characterized as purple colonies) on the modified-MRS (mMRS) medium. Metabolites such as lactic acid and diacetyl production of the strain F2 were also investigated using chromatography and found to be 10.07 and 0.05 µg/mL, respectively. The peptides of the isolate's cell-free supernatant were determined to be ∼80 kDa, and finally, the plasmid isolated from the strain F2 was identified as L. plantarum strain KLDS1.0386 plasmid p4, which may be responsible for some characteristic properties, such as antimicrobial peptide production of the strain.

Antibiotic resistant Escherichia coli strains in healthy pets from Tamaulipas, Mexico.

Antibiotic resistance constitutes a significant public health challenge, with diverse reservoirs of resistant bacteria playing pivotal roles in their dissemination. Among these reservoirs, pets are carrying antibiotic-resistant strains. The objective of this study was to assess the resistance profiles of Escherichia coli, and the prevalence of extended-spectrum ß-lactamase (ESBL) producing E. coli strains in dogs and cats from Tamaulipas, Mexico. A total of 300 stool samples (150 dogs and 150 cats) from healthy pets were subjected to analysis. Antibiotic susceptibility testing and the identification of ESBLs were carried out by disc diffusion method. The presence of resistance genes, class 1, 2, and 3 integrons (intI1, intI2, and intI3) and phylogroups was determined by PCR analysis. The findings reveal that 42.6% (128/300) of the strains exhibited resistance to at least one of the eight antibiotics assessed, and 18.6% (56/300) demonstrated multidrug resistance (MDR), that distributed across 69 distinct resistance patterns. Altogether 2.6% of E. coli strains (8/300) were confirmed as TEM and CTX-M type ESBL producers. These outcomes underscore the roles of dogs and cats in Tamaulipas as reservoirs for the dissemination of MDR and/or ESBL strains. The results underscore the necessity for conducting prevalence studies on ESBL-producing E. coli, forming a foundation for comprehending the present scenario and formulating strategies for the control and mitigation of this issue.

Development and characterization of bilayer chitosan/alginate cling film reinforced with essential oil based nanocomposite for red meat preservation.

With a goal to finding suitable alternatives to plastic packaging in the food industry, we developed a multifunctional bio-based active packaging film to enhance the shelf life of red meat. A chitosan/alginate (Chi + Alg) bilayer film was developed through layer-by-layer (LBL) assembly and an active material i.e. lemongrass nanoemulsion with silver nanoparticles-based nanocomposite (NC1) was loaded into the alginate layer to improve the quality of the bio-based film (Chi + Alg + NC1). The Chi + Alg + NC1 film was characterized in terms of its microstructure, mechanical strength, thermal stability, and antimicrobial activity. Scanning electron microscopy (SEM) revealed a film (22.5 ± 1.44 µm thickness) with a smooth and even surface and a cross-sectional structure. The incorporation of NC1 improves the quality of the film by enhancing its mechanical strength and thermal stability. FT-IR spectra showed the successful interaction between chitosan and alginate in the LBL assembly and the incorporation of NC1 in the alginate layer. The red meat preservation test demonstrated that the shelf life improved when the meat was covered with the fabricated bio-based film. The color of the meat was retained for up to 7 days compared to that of the control (Chi alone and Chi + Alg). Additionally, a reduction in the microbial count in the Chi + Alg + NC1 film was observed, corroborating the shelf-life improvement. In addition to its inherent antimicrobial properties, NC1 induced hydrophilic properties to the film, which further aids in its antimicrobial activity against E. coli. These findings suggest that Chi + Alg + NC1 film could be a potent alternative to plastic packaging and can be used as a cling film to prolong the shelf life of red meat.

Dual effect of pH and X-ray irradiation on properties of gelatin/trans-cinnamaldehyde-based composite films for sustainable packaging.

Gelatin (Gel) based water-insoluble films with antimicrobial properties were developed by the green method using trans-cinnamaldehyde (TCA) and low-energy X-ray irradiation as dual crosslinkers. The Gel/TCA composite films (GTCF) were prepared at different pH (4, 6, 8, and 10) and crosslinked by incorporating 5 % (w/w, based on Gel) TCA and X-ray irradiation (350 kV and 11.4 mA) with doses of 0, 5, 10 and 15 kGy. The presence of TCA in GTCF forms dense, flexible, and strong films when exposed to X-ray irradiation. The GTCF at pH 6, incorporated with 5 wt% TCA and irradiated with 10 kGy X-ray, displayed the highest degree of crosslinking (DOC) (93.4 ± 3.4 %), tensile strength, excellent UV-barrier (> 99.9 %), antimicrobial (inhibitory capacity of >50 %), and water vapor permeability (4.1 ± 0.6 g.mm/m2.day. kPa), and low solubility in water (0.5 ± 0.3 %), and oxygen permeability. The GTCF, crosslinked with X-ray irradiation, has multifunctional properties and strong potential in the sustainable packaging industry to augment the shelf life of food and reduce food waste. To the best of our information, this is the first and novel report investigating the effects of pH on the properties of GTCF crosslinked with X-ray.

Antimicrobial potential of Streptomyces sp. NP73 isolated from the forest soil of Northeast India against multi-drug resistant Escherichia coli.

This study reports the isolation and characterization of a Streptomyces sp. from soil, capable of producing bioactive secondary metabolites active against a variety of bacterial human pathogens. We targeted the antimicrobial activity against Escherichia coli ATCC-BAA 2469, a clinically relevant strain of bacteria harbouring resistance genes for carbapenems, extended spectrum beta-lactams, tetracyclines, fluoroquinones, etc. Preliminary screening using the spot inoculation technique identified Streptomyces sp. NP73 as the potent strain among the 74 isolated Actinomycetia strain. 16S rRNA gene and whole genome sequencing (WGS) confirmed its taxonomical identity and helped in the construction of the phylogenetic tree. WGS revealed the predicted pathways and biosynthetic gene clusters responsible for producing various types of antibiotics including the isolated compound. Bioactivity guided fractionation and chemical characterization of the active fraction, carried out using liquid chromatography, gas chromatography-mass spectrometry, infra-red spectroscopy, and nuclear magnetic resonance spectroscopy, led to the tentative identification of the active compound as Pyrrolo[1,2-a] pyrazine-1,4-dione, hexahydro-, a diketopiperazine molecule. This compound exhibited excellent antimicrobial and anti-biofilm properties against E. coli ATCC-BAA 2469 with an MIC value of 15.64 µg ml-1, and the low cytotoxicity of the compound identified in this study provides hope for future drug development.

Enhancing antibiotic prescribing practices at the main military training hospital of Tunis: A study of pharmaceutical interventions.

BACKGROUND: Bacterial infections have historically posed significant challenges until the discovery of antibiotics, which revolutionized infectious disease treatment. However, bacterial adaptation mechanisms over time have led to increased antimicrobial resistance, necessitating judicious antibiotic use. OBJECTIVES: This study aims to comprehensively analyze pharmaceutical interventions related to antibiotic prescriptions governed by antibiotic order forms to identify and rectify medication errors, optimizing antibiotic prescribing practices. MATERIAL AND METHODS: Approval for this research was obtained from the institutional review board of the Main Military Training Hospital of Tunis, Tunisia. A retrospective study was conducted at the main military training hospital of Tunis over 4 months. Pharmaceutical validation of antibiotic prescriptions through antibiotic order forms was conducted by a pharmacy resident. Pharmaceutical interventions were initiated upon detection of errors, and patient records were accessed through institutional software. RESULTS: Out of 1100 prescription forms analyzed, 41 pharmaceutical interventions were conducted for 7 antibiotics. Twenty-four percent of all interventions were related to antibiotic order forms, with the intensive care unit accounting for the highest number of errors. Under-dosage and prescription errors were common. CONCLUSION: Our pharmaceutical interventions related to antibiotic order forms are crucial for optimizing antibiotic therapy. Feedback mechanisms to healthcare teams are essential for enhancing prescription quality and patient care outcomes. Ongoing surveillance and improvement efforts are necessary to address medication errors and enhance antimicrobial stewardship.

Overcoming China's animal waste disposal challenge brought by elevated levels of veterinary antimicrobial residues and antimicrobial resistance.

Direct application of animal waste on farmlands was banned in China recently, rendering organic fertilizer production a sound solution for disposing of animal manures and recycling their materials and nutrients. Due to the overuse of antimicrobials in livestock and poultry farms, manure-based organic fertilizers often contain elevated residues of antimicrobials and abundant antimicrobial resistance genes. Land application of such products has caused significant concerns on the environmental pollution of antimicrobials, and the transmission and development of antimicrobial resistance (AMR), which is a major global health challenge. China's recent attempt to restrict the contents of antimicrobial residues in organic fertilizers encountered strong resistance from the industry as it would hinder the utilization of animal manures as a raw material. Reducing and even eliminating the use of antimicrobials in animal farms is the ultimate solution to the challenge of manure disposal posed by the elevated levels of antimicrobial residues and AMR. Phasing out the non-therapeutic use of antimicrobials, developing substitutes of antimicrobials, enhancing animal welfare in farms, promoting diversification of animal farms, and developing antimicrobial removal and disinfection technologies for animal waste are recommended to improve the veterinary antimicrobial stewardship and manure management in China's animal agriculture. These concerted measures would enhance the sustainability of crop and animal farming systems in China and mitigate the impact of antimicrobials and AMR to agro-environmental quality and human health.

Adaptations to cationic biocide exposure differentially influence virulence factors and pathogenicity in Pseudomonas aeruginosa.

Cationic biocides (CBs), which include quaternary ammonium compounds (QACs), are employed to mitigate the spread of infectious bacteria, but resistance to such surface disinfectants is rising. CB exposure can have profound phenotypic implications that extend beyond allowing microorganisms to persist on surfaces. Pseudomonas aeruginosa is a deadly bacterial pathogen that is intrinsically tolerant to a wide variety of antimicrobials and is commonly spread in healthcare settings. In this study, we pursued resistance selection assays to the QAC benzalkonium chloride and quaternary phosphonium compound P6P-10,10 to assess the phenotypic effects of CB exposure in P. aeruginosa PAO1 and four genetically diverse, drug-resistant clinical isolates. In particular, we sought to examine how CB exposure affects defensive strategies and the virulence-associated "offensive" strategies in P. aeruginosa. We demonstrated that development of resistance to BAC is associated with increased production of virulence-associated pigments and alginate as well as pellicle formation. In an in vivo infection model, CB-resistant PAO1 exhibited a decreased level of virulence compared to wild type, potentially due to an observed fitness cost in these strains. Taken together, these results illustrate the significant consequence CB resistance exerts on the virulence-associated phenotypes of P. aeruginosa.

Isolation, characterization and antimicrobial activity study of Thymus vulgaris.

Herbal medicines are important for ensuring sustainable development goals (SDGs) in healthcare, particularly in developing countries with high rates of antimicrobial resistance (AMR) and little access to medical facilities. Thymus vulgaris is a widely used herbal medicinal plant known for its secondary metabolites and antimicrobial properties. The present study involved a comprehensive examination of the isolation, characterization, and antibacterial activity of Thymus vulgaris obtained from Ethiopia. The aerial part of the plant Thymus vulgaris was successively extracted with hexane, chloroform, and methanol based on differences in polarity. Phytochemical screening tests conducted against hexane, chloroform and MeOH crude extracts indicated the presence of some secondary metabolites. Based on the thin-layer chromatography tests, the chloroform extract was subjected to column chromatography, yielding Tv-2 compounds, namely 5-isopropyl-2-methylphenol. The structures of the compounds were elucidated via spectroscopic methods (UV-Vis, FT-IR and NMR). We investigated the antibacterial properties of hexane crude extract, chloroform crude extract, MeOH crude extract, and isolated fractions derived from T. vulgaris against various bacterial strains. This study contributes to a better understanding of the bioactive components present in Thymus vulgaris crude extracts and their potential role in tackling microbial infections.

An Evidence-Based Serious Game App for Public Education on Antibiotic Use and Resistance: Randomized Controlled Trial.

BACKGROUND: The misuse and overuse of antibiotics accelerate the development of antimicrobial resistance (AMR). Serious games, any form of games that serve a greater purpose other than entertainment, could augment public education above ongoing health promotion efforts. Hence, we developed an evidence-based educational serious game app-SteWARdS Antibiotic Defence-to educate players on good antibiotic use practices and AMR through a game quest comprising 3 minigames and interaction with the nonplayer characters. OBJECTIVE: We aimed to evaluate the effectiveness of the SteWARdS Antibiotic Defence app in improving the knowledge of, attitude toward, and perceptions (KAP) of appropriate antibiotic use and AMR among the public in Singapore. METHODS: We conducted a 2-arm parallel randomized controlled trial, recruiting visitors aged 18-65 years from 2 polyclinics in Singapore. Intervention group participants had to download the SteWARdS Antibiotic Defence app (available only in English and on the Android platform) on their smartphones and complete the quest in the app. Participants took half a day to 2 weeks to complete the quest. The control group received no intervention. Knowledge questions on antibiotic use and AMR (11 binary questions) were self-administered at baseline, immediately after the intervention, and 6-10 weeks post intervention, while attitudes and perception questions (14 three-point Likert-scale questions) were self-administered at baseline and 6-10 weeks post intervention. We also collected participants' feedback on app usage. RESULTS: Participants (n=348; intervention: n=142, control: n=206) had a mean age of 36.9 years. Intervention group participants showed a statistically significant improvement in mean knowledge score (effect size: 0.58 [95% CI 0.28-0.87]) compared with controls after accounting for age, educational level, and exposure to advertisements on antibiotics and AMR. Intervention participants also showed a statistically significant improvement in mean attitude-perception scores (effect size: 0.98 (95% CI 0.44-1.52)) after adjusting for marital status and race. A majority of participants agreed that the "SteWARdS Antibiotic Defence" app improved their awareness on antibiotic use (135/142, 95.1%) and AMR (136/142, 95.8%). About 73.9% (105/142) of the participants agreed that the app is easy to use, 70.4% (100/142) agreed that the app was enjoyable, and 85.2% (121/142) would recommend the app to others. CONCLUSIONS: Our educational serious game app improves participants' KAP on appropriate antibiotic use and AMR. Public education apps should be engaging, educational, easy to use, and have an attractive user interface. Future research should assess the effectiveness of interventions in facilitating long-term knowledge retention and long-lasting behavioral change. TRIAL REGISTRATION: ClinicalTrials.gov NCT05445414; https://clinicaltrials.gov/ct2/show/NCT05445414. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): RR2-10.2196/45833.

Dynamics of antimicrobial resistance and genomic characteristics of foodborne Vibrio spp. in Southern China (2013-2022).

Vibrio spp., known as significant marine pathogens, have become more prevalent due to global warming. Antibiotics released into the environment drive Vibrio resistance. The increasing consumption of seafood leads to more interactions between Vibrio and humans. Despite this concerning trend, there remains a lack of large-scale surveillance for Vibrio contamination across various types of food. This study isolated 4027 Vibrio strains, primarily comprising V. parahaemolyticus and V. alginolyticus, in 3581 fresh shrimp and meat products from 2013 to 2022. The Vibrio strains showed increased resistance to important antibiotics, especially ß-lactams used to treat foodborne bacterial infections. Whole genome sequencing of 591 randomly chosen strains showed a strong correlation between antibiotic resistance and genotypes in Vibrio. Notably, various ESBL genes have evolved over the past 8 years, with blaVEBs being the most dominant. Additionally, carbapenemase genes, such as blaNDM-1, have become increasingly prevalent in recent years. Various mobile genetic elements, including IncQ and IncA/C plasmids, recoverable in Vibrio, facilitate the transmission of crucial ß-lactamase genes. These data provide insights into the evolutionary traits of antimicrobial resistance in foodborne Vibrio strains over a decade. Policymakers should consider these findings when devising appropriate strategies to combat bacterial antimicrobial resistance and safeguard human health.

Drivers of virulence and antimicrobial resistance in Gram-negative bacteria in different settings: A genomic perspective.

The human gut presents a complex ecosystem harboring trillions of microorganisms living in close association with each other and the host body. Any perturbation or imbalance of the normal gut microbiota may prove detrimental to human health. Enteric infections and treatment with antibiotics pose major threats to gut microbiota health. Recent genomics-driven research has provided insights into the transmission and evolutionary dynamics of major enteric pathogens such as Escherichia coli, Klebsiella pneumoniae, Vibrio cholerae, Helicobacter pylori and Salmonella spp. Studies entailing the identification of various dominant lineages of some of these organisms based on artificial intelligence and machine learning point to the possibility of a system for prediction of antimicrobial resistance (AMR) as some lineages have a higher propensity to acquire virulence and fitness advantages. This is pertinent in the light of emerging AMR being one of the immediate threats posed by pathogenic bacteria in the form of a multi-layered fitness manifesting as phenotypic drug resistance at the level of clinics and field settings. To develop a holistic or systems-level understanding of such devastating traits, present methodologies need to be advanced with the high throughput techniques integrating community and ecosystem/niche level data across different omics platforms. The next major challenge for public health epidemiologists is understanding the interactions and functioning of these pathogens at the community level, both in the gut and outside. This would provide new insights into the dimensions of enteric bacteria in different environments and niches and would have a plausible impact on infection control strategies in terms of tackling AMR. Hence, the aim of this review is to discuss virulence and AMR in Gram-negative pathogens, the spillover of AMR and methodological advancements aimed at addressing it through a unified One Health framework applicable to the farms, the environment, different clinical settings and the human gut.

Antimicrobial prescription pattern and appropriateness for respiratory tract infection in outpatients: a systematic review and meta-analysis.

BACKGROUND: Millions of people die every year as a result of antimicrobial resistance worldwide. An inappropriate prescription of antimicrobials (e.g., overuse, inadequate use, or a choice that diverges from established guidelines) can lead to a heightened risk of antimicrobial resistance. This study aimed to determine the rate and appropriateness of antimicrobial prescriptions for respiratory tract infections. METHODS: This review was conducted in accordance with the PRISMA guidelines. Web of Science, PubMed, ProQuest Health and Medicine, and Scopus were searched between October 1, 2023, and December 15, 2023, with no time constraints. Studies were independently screened by the first author and the co-authors. We included original studies reporting antimicrobial prescription patterns and appropriateness for respiratory tract infections. The quality of included studies' was assessed via the Joanna Briggs Institute's Critical Appraisal Checklists for Cross-Sectional Studies. The assessment of publication bias was conducted using a funnel plot and Egger's regression test. A random effect model was employed to estimate the pooled antibiotic prescribing and inappropriate rates. Subgroup analysis was conducted by country, study period, data source, and age group. RESULTS: Of the total 1220 identified studies, 36 studies were included in the review. The antimicrobial prescribing rate ranged from 25% (95% CI 0.24-0.26) to 90% (95% CI 0.89-0.91). The pooled antimicrobial prescription rate was 66% (95% CI 0.57 to 0.73). Subgroup analysis by region revealed that the antimicrobial prescription rate was highest in Africa (79%, 95% CI 0.48-0.94) and lowest in Europe (47%, 95% CI 0.32-0.62). Amoxicillin and amoxicillin-clavulanate antimicrobials from the Access group, along with azithromycin and erythromycin from the Watch group, were the most frequently used antimicrobial agents. This study revealed that the major reasons for antimicrobial prescription were acute bronchitis, pharyngitis, sinusitis, and the common cold. The pooled inappropriate antimicrobial prescription rate was 45% (95% CI 0.38-0.52). Twenty-eight of the included studies reported that prescribing antimicrobials without proper indications was the main cause of inappropriate antimicrobial prescriptions. Additionally, subgroup analysis by region showed a higher inappropriate antimicrobial prescription rate in Asia at 49% (95% CI 0.38-0.60). The result of the funnel plot and Egger's tests revealed no substantial publication bias (Egger's test: p = 0.268). CONCLUSION: The prescribing rate and inappropriate use of antimicrobials remain high and vary among countries. Further studies should be conducted to generate information about factors contributing to unnecessary antimicrobial prescriptions in outpatients. SYSTEMATIC REVIEW REGISTRATION: Systematic review registration: CRD42023468353.

Antimicrobial properties of the edible pink oyster mushroom, Pleurotus eous: In-vivo and in-vitro studies.

In recent times, there has been a notable surge in the investigation of new antibiotic substances derived from natural origins. Pleurotus eous is an edible mushroom that has various useful bioactive substances and therapeutic properties, including antimicrobial activity. The present study aims to evaluate the antimicrobial efficacy of the methanolic extract of P. eous (MEPE) through in vitro method. Notably, S. aureus demonstrated the highest susceptibility to MEPE, prompting further investigation into its antibacterial mechanisms via scanning electron microscopy (SEM), membrane integrity, and permeability assays. The in-vivo antibacterial effect of MEPE against S. aureus was also assessed, including analysis of bacterial burden in organs, hematological profiles, and cytokine profiles. Detailed phytochemical analyses of MEPE were conducted using GC-MS. Results revealed MEPE's significant (p < 0.05) efficacy against Gram-positive bacteria, particularly S. aureus (77.56 ± 0.4 µg/mL and 34 ± 6.9 µg/ml in turbidometric and viable cell count assays, respectively). Moreover, membrane permeability significantly increased in 60.32 % of S. aureus isolates following treatment with MEPE. Additionally, mice receiving MEPE exhibited decreased levels of TNF-α, IL-1ß, and IL-6, suggesting its potential in combating S. aureus infection in animal models.

Effect of anaerobic digestion on pathogens and antimicrobial resistance in the sewage sludge.

Antimicrobial resistance (AMR) is recognized as a global threat. AMR bacteria accumulate in sewage sludge however, knowledge on the persistence of human pathogens and AMR in the sludge line of the wastewater treatment is limited. Sludge can be used, with or without additional treatment, as fertilizer in agricultural fields. The aim of this study is to obtain knowledge about presence of human pathogens and AMR in the sewage sludge, before and after the anaerobic digestion (AD) applying innovative combinations of methods. Fifty sludge samples were collected. Cultivation methods combined with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and Antibiotic Susceptibility Test (AST) were used obtaining knowledge about the microbial community, pathogens, and antibiotic resistant bacteria while the droplet digital Polymerase Chain Reaction (ddPCR) was performed to detect most common AMR genes. In total, 231 different bacterial species were identified in the samples. The most abundant species were spore-forming facultative anaerobic bacteria belonging to Bacillus and Clostridium genera. The AD causes a shift in the microbial composition of the sludge (p = 0.04). Seven pathogenic bacterial species constituting 188 colonies were isolated and tested for susceptibility to Clindamycin, Meropenem, Norfloxacin, Penicillin G, and Tigecycline. Of the Clostridium perfringens and Bacillus cereus isolates 67 and 50 %, respectively, were resistant to Clindamycin. Two B. cereus and two C. perfringens isolates were also resistant to other antibiotics showing multidrug resistance. ARGs (blaOXA, blaTEM, ermB, qnrB, tet(A)-(W), sulI-II) were present at 7-8 Log gene copies/kg of sludge. AD is the main driver of a reduction of some ARGs (1 Log) but resistant bacteria were still present. The results showed the usefulness of the integration of the proposed analytical methods and suggest a decrease in the risk of presence of cultivable pathogens including resistant isolates after AD but a persistent risk of ARGs' horizontal transmission.