Biofilm Antibiotic Resistance Research Articles

Artificial Phages with Biocatalytic Spikes for Synergistically Eradicating Antibiotic-Resistant Biofilms.

Antibiotic-resistant pathogens have become a global public health crisis, especially biofilm-induced refractory infections. Efficient, safe, and biofilm microenvironment (BME)-adaptive therapeutic strategies are urgently demanded to combat antibiotic-resistant biofilms. Here, inspired by the fascinating biological structures and functions of phages, we propose the de novo design of a spiky Ir@Co3O4 particle to serve as an artificial phage for synergistically eradicating antibiotic-resistant Staphylococcus aureus biofilms. Benefiting from the abundant nanospikes and highly active Ir sites, the synthesized artificial phage can simultaneously achieve efficient biofilm accumulation, extracellular polymeric substance (EPS) penetration, and superior BME-adaptive reactive oxygen species (ROS) generation, thus facilitating the in situ ROS delivery and enhancing the biofilm eradication. Moreover, metabolomics found that the artificial phage obstructs the bacterial attachment to EPS, disrupts the maintenance of the BME, and fosters the dispersion and eradication of biofilms by down-regulating the associated genes for the biosynthesis and preservation of both intra- and extracellular environments. Our in vivo results demonstrate that the artificial phage can treat the biofilm-induced recalcitrant infected wounds equivalent to vancomycin. We suggest that the design of this spiky artificial phage with synergistic "penetrate and eradicate" capability to treat antibiotic-resistant biofilms offers a new pathway for bionic and non-antibiotic disinfection. This article is protected by copyright. All rights reserved.

Functionalized chitosan based antibacterial hydrogel sealant for simultaneous infection eradication and tissue closure in ocular injuries

Management of infections at ocular injury often requires prolonged and high dose of antibiotic, which is associated with challenges of antibiotic resistance and bacterial biofilm formation. Tissue glues are commonly used for repairing ocular tissue defects and tissue regeneration, but they are ineffective in curing infection. There is a critical need for antibacterial ocular bio-adhesives capable of both curing infection and aiding wound closure. Herein, we present the development of an imine crosslinked N-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride (HTCC)‑silver chloride nanocomposites (QAm1-Agx) and poly-dextran aldehyde (PDA) based bactericidal sealant (BacSeal). BacSeal exhibited potent bactericidal activity against a broad spectrum of bacteria including their planktonic and stationary phase within a short duration of 4 h. BacSeal effectively reduced biofilm-embedded MRSA and Pseudomonas aeruginosa by ∼99.99 %. In ex-vivo human cornea infection model, BacSeal displayed ∼99 % reduction of ocular infection. Furthermore, the hydrogel exhibited excellent sealing properties by maintaining ocular pressure up to 75 mm-Hg when applied to human corneal trauma. Cytotoxicity assessment and hydrogel-treated human cornea with a retained tissue structure, indicate its non-toxic nature. Collectively, BacSeal represents a promising candidate for the development of an ocular sealant that can effectively mitigate infections and may assist in tissue regeneration by sealing ocular wounds.

Plasmid-mediated transfer of antibiotic resistance genes and biofilm formation in a simulated drinking water distribution system under chlorine pressure

The effects of disinfectants and plasmid-based antibiotic resistance genes (ARGs) on the growth of microorganisms and the plasmid-mediated transfer of ARGs in the water and biofilm of the drinking water distribution system under simulated conditions were explored. The heterotrophic plate count of the water in reactors with 0.1 mg/L NaClO and NH2Cl was higher than in the control groups. There was no similar phenomenon in biofilm. In the water of reactors containing NaClO, the aphA and bla genes were lower than in the antibiotic resistant bacteria group, while both genes were higher in the water of reactors with NH2Cl than in the control group. Chloramine may promote the transfer of ARGs in the water phase. Both genes in the biofilm of the reactors containing chlorine were lower than the control group. Correlation analysis between ARGs and water quality parameters revealed that the copy numbers of the aphA gene were significantly positively correlated with the copy numbers of the bla gene in water and significantly negatively correlated in biofilm (p < 0.05). The results of the sequencing assay showed that bacteria in the biofilm, in the presence of disinfectant, were primarily Gram-negative. 1.0 mg/L chlorine decreased the diversity of the community in the biofilm. The relative abundance of some bacteria that may undergo transfer increased in the biofilm of the reactor containing 0.1 mg/L chlorine.

Selective antibacterial and antibiofilm activity of chlorinated hemicyanine against gram-positive bacteria

Antibiotic-free therapies are highly needed due to the limited success of conventional approaches especially against biofilm related infections. In this direction, antimicrobial phototherapy, either in the form of antimicrobial photothermal therapy (aPTT) or antimicrobial photodynamic therapy (aPDT), have appeared to be highly promising candidates in recent years. These are local and promising approaches for antibiotic resistant bacterial infections and biofilms. Organic small photosensitizers (PSs) are extensively preferred in antimicrobial phototherapy applications as they offer a great opportunity to combine therapeutic action (aPTT, aPDT or both) with fluorescence imaging on a single molecule. In this study, the bactericidal effect of cationic chlorinated hemicyanine (Cl-Hem)-based type I PS, which can function as a dual aPDT/aPTT agent, was investigated on both planktonic cells and biofilms of different gram-positive (E. faecalis and S. epidermidis) and gram-negative bacteria (P. aeruginosa and K. pneumoniae) with and without 640 nm laser irradiation. Cl-Hem was shown to induce a selective phototheranostic activity against gram-positive bacteria (E. faecalis and S. epidermidis). Cl-Hem exhibited both dose and laser irradiation time dependent bactericidal effect on planktonic and biofilms of S. epidermidis. These results clearly showed that highly potent Cl-Hem can treat resistant microbial infections, while allowing fluorescence detection at the same time. High biofilm reduction observed with combined aPDT/aPTT action of Cl-Hem together with its non-cytotoxic nature points out that Cl-Hem is a promising PS for antibacterial and antibiofilm treatments.

Rabbits as a Reservoir of Multidrug-Resistant Escherichia coli: Clonal Lineages and Public Health Impact.

Escherichia coli, including extended-spectrum β-lactamases (ESBL)-producing strains, poses a global health threat due to multidrug resistance, compromising food safety and environmental integrity. In industrial settings, rabbits raised for meat have the highest consumption of antimicrobial agents compared to other food-producing animals. The European Union is facing challenges in rabbit farming as rabbit consumption declines and antibiotic-resistant strains of E. coli cause enteric diseases. The aim of this study was to investigate the antibiotic resistance profile, genetic diversity, and biofilm formation in cefotaxime-resistant E. coli strains isolated from twenty rabbit farms in Northern Portugal to address the effect of the pressing issue of antibiotic resistance in the rabbit farming industry. Resistance to critically antibiotics was observed, with high levels of resistance to several categories, such as tetracycline, ampicillin, aztreonam, and streptomycin. However, all isolates were susceptible to cefoxitin and imipenem. Multidrug resistance was common, with strains showing resistance to all antibiotics tested. The blaCTX-M variants (blaCTX-3G and blaCTX-M9), followed by the tetracycline resistance genes, were the most frequent resistance genes found. ST10 clones exhibiting significant resistance to various categories of antibiotics and harboring different resistance genes were detected. ST457 and ST2325 were important sequence types due to their association with ESBL-E. coli isolates and have been widely distributed in a variety of environments and host species. The strains evaluated showed a high capacity for biofilm formation, which varied when they were grouped by the number of classes of antibiotics to which they showed resistance (i.e., seven different classes of antibiotics, six classes of antibiotics, and three/four/five classes of antibiotics). The One Health approach integrates efforts to combat antimicrobial resistance in rabbit farming through interdisciplinary collaboration of human, animal, and environmental health. Our findings are worrisome and raise concerns. The extensive usage of antibiotics in rabbit farming emphasizes the urgent need to establish active surveillance systems.

Comparative Analysis of Antibiotic Resistance and Biofilm Characteristics of Two Major Enterococcus Species from Poultry Slaughterhouses in South Korea.

The spread of antibiotic-resistant Enterococcus in the poultry industry poses significant public health challenges due to multidrug resistance and biofilm formation. We investigated the antibiotic resistance profiles and biofilm characteristics of E. faecalis and E. faecium isolates from chicken meat in poultry slaughterhouses in South Korea. Ninety-six isolates (forty-eight each of E. faecalis and E. faecium) were collected between March and September 2022. Both species were analyzed using MALDI-TOF, PCR, antibiotic susceptibility testing, and biofilm assays. A high level of multidrug resistance was observed in E. faecalis (95.8%) and E. faecium (93.8%), with E. faecium exhibiting a broader range of resistance, particularly to linezolid (52.1%) and rifampicin (47.9%). All E. faecalis isolates formed biofilm in vitro, showing stronger biofilm formation than E. faecium with a significant difference (p < 0.001) in biofilm strength. Specific genes (cob, ccf, and sprE) were found to be correlated with biofilm strength. In E. faecium isolates, biofilm strength was correlated with resistance to linezolid and rifampicin, while a general correlation between antibiotic resistance and biofilm strength was not established. Through analysis, correlations were noted between antibiotics within the same class, while no general trends were evident in other analyzed factors. This study highlights the public health risks posed by multidrug-resistant enterococci collected from poultry slaughterhouses, emphasizing the complexity of the biofilm-resistance relationship and the need for enhanced control measures.

Antimicrobial resistance of Pseudomonas aeruginosa: navigating clinical impacts, current resistance trends, and innovations in breaking therapies.

Pseudomonas aeruginosa, a Gram-negative bacterium, is recognized for its adaptability and opportunistic nature. It poses a substantial challenge in clinical settings due to its complicated antibiotic resistance mechanisms, biofilm formation, and capacity for persistent infections in both animal and human hosts. Recent studies revealed a potential zoonotic transmission of P. aeruginosa between animals, the environment, and human populations which highlights awareness of this microbe. Implementation of the One Health approach, which underscores the connection between human, animal, and environmental health, we aim to offer a comprehensive perspective on the current landscape of P. aeruginosa management. This review presents innovative strategies designed to counteract P. aeruginosa infections. Traditional antibiotics, while effective in many cases, are increasingly compromised by the development of multidrug-resistant strains. Non-antibiotic avenues, such as quorum sensing inhibition, phage therapy, and nanoparticle-based treatments, are emerging as promising alternatives. However, their clinical application encounters obstacles like cost, side effects, and safety concerns. Effectively addressing P. aeruginosa infections necessitates persistent research efforts, advancements in clinical development, and a comprehension of host-pathogen interactions to deal with this resilient pathogen.

Comparative genomic analysis of Pseudomonas aeruginosa strains susceptible and resistant to carbapenems and aztreonam isolated from patients with healthcare-associated infections in a Mexican hospital.

Pseudomonas aeruginosa (PA) is an important opportunistic pathogen that causes different infections on immunocompromised patients. Within PA accessory genome, differences in virulence, antibiotic resistance and biofilm formation have been described between strains, leading to the emergence of multidrug-resistant strains. The genome sequences of 17 strains isolated from patients with healthcare-associated infections in a Mexican hospital were genomically and phylogenetically analyzed and antibiotic resistance genes, virulence genes, and biofilm formation genes were detected. Fifteen of the 17 strains were resistant to at least two of the carbapenems meropenem, imipenem, and the monobactam aztreonam. The antibiotic resistance (mexA, mexB, and oprM) and the biofilm formation (pslA and pslD) genes were detected in all strains. Differences were found between strains in accessory genome size. The strains had different sequence types, and seven strains had sequence types associated with global high risk epidemic PA clones. All strains were represented in two groups among PA global strains. In the 17 strains, horizontally acquired resistance genes to aminoglycosides and beta-lactams were found, mainly, and between 230 and 240 genes that encode virulence factors. The strains under study were variable in terms of their accessory genome, antibiotic resistance, and virulence genes. With these characteristics, we provide information about the genomic diversity of clinically relevant PA strains.

Interdisciplinary Research on Electrons and Biofilms

Interdisciplinary research on electrons and biofilms is being carried out by Assistant Professor Dr Yoshihide Tokunou, Faculty of Life and Environmental Sciences, University of Tsukuba, Japan. His work is aimed at gaining a deeper understanding of the role of electrons in bacterial society and exploring the process of bacterial extracellular electron transfer (EET) in biofilms. By combining microbiology and electrochemistry, Dr Tokunou is developing EET-based biotechnologies and improving our understanding of the physiological importance of EET in bacterial society. To achieve this, he is collaborating with various experts such as microbiologists, chemists, and engineers, including Professor Dr Nobuhiko Nomura. His current interest is in the EET process that terminates with soluble electron acceptors. By regulating the EET in biofilms respiring with oxygen, he is trying to control various biofilm-related phenomena such as host-bacteria interaction, antibiotic resistance of biofilms, biochemical production from biofilms, etc.

Antibiofilm Activity of Probiotic Strains against Aggregatibacter actinomycetemcomitans

Aggregatibacter actinomycetemcomitans is a significant agent for periodontal disease. It develops biofilms on gingival plaque and promotes chronic pro-inflammatory response to periodontal tissues. As bacterial cells within the biofilm, this pathogen successfully eluded the human immune system by preventing the uptake of biofilm cells by phagocytic cells. Antibiotic-resistant biofilms might harm the environment, increase mortality, and add to the cost of healthcare. Anti-biofilms are necessary to prevent the growth of biofilms. It has been demonstrated that probiotics change how immune system cells produce cytokines. Therefore, the efficacy of five probiotic lactobacillus strains to prevent A. actinomycetemcomitans from growing bacteria and forming biofilms was evaluated. The organisms were incubated in anaerobic conditions at 37°C for a certain period. The antimicrobial test was done using the lawn method and measuring the size of the inhibition zone produced by the probiotic spots for each probiotic strain. Tetracycline antibiotic disc was used as a positive control to compare the size of the inhibition zone. The antibiofilm activity was performed using probiotic strains at their exponential and stationary phases against the exponential phase of A. actinomycetemcomitans. Our finding showed the probiotic lactobacillus could inhibit the growth of A. actinomycetemcomitans. The highest inhibition was demonstrated by L. johnsonii NBRC 13952. Besides, all probiotic lactobacillus used for this study significantly (p&lt;0.05) inhibited the A. actinomycetemcomitans biofilm using both stationary and exponential phases. In conclusion, probiotic lactobacillus might be a potential agent that can be used to counteract the periodontal pathogen biofilm. Further study should be done to elucidate the antibiofilm mechanism, and several trials of the probiotic lactobacilli against multiple species of biofilms might be helpful in evaluating their effectiveness.

Nitrogen and phosphorus eutrophication enhance biofilm-related drug resistance in Enterococcus faecalis isolated from Water Sources

Antibiotic resistance is a critical topic worldwide with important consequences for public health. So considering the rising issue of antibiotic-resistance in bacteria, we explored the impact of nitrogen and phosphorus eutrophication on drug resistance mechanisms in Enterococcus faecalis, especially ciprofloxacin, oxytetracycline, and ampicillin. For this purpose we examined the antibiotic-resistance genes and biofilm formation of Enterococcus faecalis under different concentration of nitrogen and phosphorus along with mentioned antibiotics. Mesocosms were designed to evaluate the impact of influence of eutrophication on the underlying mechanism of drugn resistence in Enterococcus faecalis. For this purpose, we explored the potential relation to biofilm formation, adhesion ability, and the expression levels of the regulatory gene fsrA and the downstream gene gelEI. Our results demonstrated that the isolates of all treatments displayed high biofilm forming potential, and fsrA and gelE genes expression. Additionally, the experimental group demonstrated substantially elevated Enterococcus faecalis gelE expression. Crystal violet staining was applied to observe biofilm formation during bacterial development phase and found higher biofilm formation. In conclusion, our data suggest that E. faecalis resistance to ciprofloxacin, oxytetracycline, and ampicillin is related to biofilm development. Also, the high level of resistance in Enterococcus faecalis is linked to the expression of the fsrA and gelE genes. Understanding these pathways is vital in tackling the rising problem of bacterial resistance and its potential effect on human health.

Prevalence, extended-spectrum β-lactamase and biofilm production ability ofEscherichia coli isolated from buffalo mastitis

This study aimed to determine the prevalence, antibiotic resistance pattern, extended-spectrum, beta-lactamase production and biofilm forming ability of isolated E. coli strains from buffaloes mastitis milk. Out of 549 bacterial isolates from mastitis milk of buffaloes (n, animal level= 472) between 2019 and 2022, a total of 43 E. coli strains were isolated with an overall prevalence of 9.11% at animal level. Prevalence of E coli was high in unorganised buffalo herd (11.36%) from villages of Farmer FIRST project (ICAR-FFP) compared with organised buffalo farms (6.73%). The highest resistance was against Penicillin 43 (100%) followed by Ceftriaxone 18 (41.86%), Amoxycillin/Sulbactam 8 (18.60%) and Enrofloxacin 7 (16.27%). Additionally, all were sensitive to gentamycin 43 (100%) followed by Cefoperazone/Sulbactam 34 (79.06%). Cephalosporins are frequently used antibiotics to treat bovine mastitis. However, their therapeutic effectiveness is being compromised by bacterial resistant to β-lactams. In present study, a total of 32 (6.78% %) extended-spectrum β-lactamase (ESBL) producing E coli were isolated from mastitic buffalo milk (n=43/472). In total, 17 (39.5%) isolates were biofilm producers by microtiter-plate method. There was statistically non-significant relationship between biofilm production and antibiotic resistance as well as between ESBL production and biofilm formation in E coli strains. Present study demonstrated a high occurrence of ESBL and biofilm producing E. coli in buffalo mastitis milk, implementing a significant challenge to treat mastitis in buffaloes, necessitates judicious use of antimicrobials and to explore potential therapeutic agents as substitutes forantibiotics to treat bovine mastitis effectively.

Synergistic Effect of Thymol-Ciprofloxacin Combination on Planktonic Cells and Biofilm of Pseudomonas aeruginosa.

Pseudomonas aeruginosa is an opportunistic bacteria causing severe and life-threatening infections in individuals with weakened immune systems. P. aeruginosa forms antibiotic-resistant biofilms, rendering it challenging to treat; hence, alternate therapies are required to eliminate it. Treatment of infections using a combination of drugs is gaining momentum to combat drug-resistant pathogens, including P.aeruginosa. This study explores the synergistic effects of Thymol in combination with Ciprofloxacin, Amikacin and Colistin against planktonic cells and biofilm of P.aeruginosa. Thymol in combination with Ciprofloxacin yields the fractional inhibitory concentration index values 0.156 and 0.375 in P.aeruginosa strains, GC14 and ATCC 9027, respectively, highlighting a robust synergistic effect on both the planktonic and biofilm of P.aeruginosa. The results showed that Thymol (512μg/mL) and Ciprofloxacin (0.125μg/mL) were the most effective combination with 95 and 93.5% total biofilm inhibition in GC14 and PA27, respectively, compared to the Thymol (512μg/mL) and Ciprofloxacin (0.125μg/mL) alone. Our findings suggest that the combinations of Thymol and Ciprofloxacin may be a potential therapeuticstrategy to address the issue of infections caused by P.aeruginosa biofilms.

In vitro evaluation of biofilm phenotypic and genotypic characteristics among clinical isolates of Pseudomonas aeruginosa in Hamadan, West of Iran.

Due to high antimicrobial resistance and biofilm-forming ability, Pseudomonas aeruginosa is one of the seriously life-threatening agents causing chronic and nosocomial infections. This study was performed to determine the antibiotic resistance pattern, biofilm formation, and frequency of biofilm-related genes in P. aeruginosa strains. In total, 123 P. aeruginosa isolates were collected from different clinical sources. Antimicrobial susceptibility testing (AST) was performed to detect multidrug-resistant P. aeruginosa (MDRPA) isolates. To evaluate the biofilm-forming isolates, the microtiter plate (MTP) method was carried out. Also, the prevalence of biofilm genotype patterns, including pslA, pslD, pelA, pelF, and algD genes, was detected by polymerases chain reaction (PCR). According to our findings, the highest resistance and susceptibility rates were found in ceftazidime with 74.7% (n = 92) and ciprofloxacin with 42.2% (n = 52), respectively. In our study, the highest level of antibiotic resistance belonged to wound isolates which meropenem had the most antibacterial activity against them. In total, 86.1% (n = 106) P. aeruginosa isolates were determined as MDRPA, of which 61.3% (n = 65) were able to form strong biofilm. The highest and lowest frequency of biofilm-related genes among biofilm producer isolates belonged to pelF with 82.1% (n = 101) and algD with 55.2% (n = 68), respectively. The findings of the conducted study indicate a significant relationship between MDRPA and biofilm genotypic/phenotypic patterns, suggesting the necessity of a careful surveillance program in hospital settings.

Biofilm tolerance, resistance and infections increasing threat of public health

Microbial biofilms can cause chronic infection. In the clinical setting, the biofilm-related infections usually persist and reoccur; the main reason is the increased antibiotic resistance of biofilms. Traditional antibiotic therapy is not effective and might increase the threat of antibiotic resistance to public health. Therefore, it is urgent to study the tolerance and resistance mechanism of biofilms to antibiotics and find effective therapies for biofilm-related infections. The tolerance mechanism and host reaction of biofilm to antibiotics are reviewed, and bacterial biofilm related diseases formed by human pathogens are discussed thoroughly. The review also explored the role of biofilms in the development of bacterial resistance mechanisms and proposed therapeutic intervention strategies for biofilm related diseases.

Recent advance on nanoparticles or nanomaterials with anti-multidrug resistant bacteria and anti-bacterial biofilm properties: A systematic review

ObjectiveWith the wide spread of Multidrug-resistant bacteria (MDR) due to the transfer and acquisition of antibiotic resistance genes and the formation of microbial biofilm, various researchers around the world are looking for a solution to overcome these resistances. One potential strategy and the best candidate to overcome these infections is using an effective nanomaterial with antibacterial properties against them. Methodsand analysis: In this study, we overview nanomaterials with anti-MDR bacteria and anti-biofilm properties. Hence, we systematically explored biomedical databases (Web of Sciences, Google Scholar, PubMed, and Scopus) to categorize related studies about nanomaterial with anti-MDR bacteria and anti-biofilm activities from 2007 to December 2022. ResultsIn total, forty-one studies were investigated to find antibacterial and anti-biofilm information about the nanomaterial during 2007–2022. According to the collected documents, nineteen types of nanomaterial showed putative antibacterial effects such as Cu, Ag, Au, Au/Pt, TiO2, Al2O3, ZnO, Se, CuO, Cu/Ni, Cu/Zn, Fe3O4, Au/Fe3O4, Au/Ag, Au/Pt, Graphene O, and CuS. In addition, seven types of them considered as anti-biofilm agents such as Ag, ZnO, Au/Ag, Graphene O, Cu, Fe3O4, and Au/Ag. ConclusionAccording to the studies, each of nanomaterial has been designed with different methods and their effects against standard strains, clinical strains, MDR strains, and bacterial biofilms have been investigated in-vitro and in-vivo conditions. In addition, nanomaterials have different destructive mechanism on bacterial structures. Various nanoparticles (NP) introduced as the best candidate to designing new drug and medical equipment preventing infectious disease outbreaks by overcome antibiotic resistance and bacterial biofilm.

Distribution and association of antimicrobial resistance and virulence characteristics in Enterococcus spp. isolates from captive Asian elephants in China.

Enterococcus spp., as an opportunistic pathogen, are widely distributed in the environment and the gastrointestinal tracts of both humans and animals. Captive Asian elephants, popular animals at tourist attractions, have frequent contact with humans. However, there is limited information on whether captive Asian elephants can serve as a reservoir of antimicrobial resistance (AMR). The aim of this study was to characterize AMR, antibiotic resistance genes (ARGs), virulence-associated genes (VAGs), gelatinase activity, hemolysis activity, and biofilm formation of Enterococcus spp. isolated from captive Asian elephants, and to analyze the potential correlations among these factors. A total of 62 Enterococcus spp. strains were isolated from fecal samples of captive Asian elephants, comprising 17 Enterococcus hirae (27.4%), 12 Enterococcus faecalis (19.4%), 8 Enterococcus faecium (12.9%), 7 Enterococcus avium (11.3%), 7 Enterococcus mundtii (11.3%), and 11 other Enterococcus spp. (17.7%). Isolates exhibited high resistance to rifampin (51.6%) and streptomycin (37.1%). 50% of Enterococcus spp. isolates exhibited multidrug resistance (MDR), with all E. faecium strains demonstrating MDR. Additionally, nine ARGs were identified, with tet(M) (51.6%), erm(B) (24.2%), and cfr (21.0%) showing relatively higher detection rates. Biofilm formation, gelatinase activity, and α-hemolysin activity were observed in 79.0, 24.2, and 14.5% of the isolates, respectively. A total of 18 VAGs were detected, with gelE being the most prevalent (69.4%). Correlation analysis revealed 229 significant positive correlations and 12 significant negative correlations. The strongest intra-group correlations were observed among VAGs. Notably, we found that vancomycin resistance showed a significant positive correlation with ciprofloxacin resistance, cfr, and gelatinase activity, respectively. In conclusion, captive Asian elephants could serve as significant reservoirs for the dissemination of AMR to humans.

Detection of Antimicrobial Resistance and Biofilm Production Among Staphylococcus pseudintermedius from Canine Skin Lesions.

Aims: Staphylococcus pseudintermedius is an opportunistic pathogen also indicated as one of the major causes of skin infections in dogs. This study aimed to identify S. pseudintermedius isolated from canine skin lesions, evaluate their antibiotic resistance profile and biofilm production ability. Methodology: Lesions from 50 rural dogs with different skin lesions were sampled after pyoderma diagnosis by private practices. Bacterial species determination was investigated and susceptibility to nine antimicrobials were determined by means of Kirby-Bauer assay. Then seven antibiotic resistance genes, including mecA, blaZ, tetK, tetM, blaSHV, blaOXA-1, and blaTEM were screened by PCR. Moreover, biofilm formation ability of the strains was determined using the microtiter plate assay along with the presence of icaADBC genes. Results: A total of 37 (74%) isolates were identified as S. pseudintermedius. All S. pseudintermedius isolates were resistant to multiple drugs. Resistance to penicillin, amoxicillin-clavulanic acid, and cefazolin were higher than other antimicrobials. All the beta-lactam-resistant isolates carried blaZ, whereas mecA was found in 6 (16.21%) of them. Among tetracycline-resistant strains, the frequency of tetK and tetM determinants were 19 (90.47%) and 21 (100%), respectively. Finally, most cefazolin-resistant strains 31 (91.89%) were positive for blaTEM gene. The rate of biofilm production ability among S. pseudintermedius isolates was 35 (94.59%). Furthermore, the presence of icaA/D genes was detected in 35 (100%) and 29 (82.85%) of S. pseudintermedius strains, respectively. Conclusion: The high rates of antimicrobial resistance and biofilm production ability among S. pseudintermedius isolates, emphasize the urgent need to use more effective infection control policies and guidelines for antimicrobial use.

Effect of ZnO nanoparticles on biofilm formation and gene expression of the toxin-antitoxin system in clinical isolates of Pseudomonas aeruginosa

BackgroundBiofilm formation by Pseudomonas aeruginosa (P. aeruginosa) is known to be characteristic of this organism. This bacterium is considered one of the most life-threatening bacteria and has been identified as a priority pathogen for research by WHO. Biofilm-producing P. aeruginosa is a concern in many parts of the world due to antibiotic resistance. Alginate also plays an important role in the biofilm formation of P. aeruginosa as well as the emergence of antibiotic resistance in biofilms. In addition, the systems of toxin-antitoxin( TA) play an important role in biofilm formation. Metal nanoparticle(NP) such as zinc oxide (ZnO) also have extensive biological properties, especially anti-biofilm properties. Therefore, this study was conducted in relation to the importance of zinc oxide nanoparticles (ZnO NPs) in biofilm formation and also the correlation of gene expression of TA systems in clinical isolates of P. aeruginosa.MethodsA total of 52 P. aeruginosa isolates were collected from burns (n = 15), UTI (n = 31), and trachea (n = 6) in hospitals in Ilam between May 2020 and October 2020. Biofilm formation was assessed using a microtiter plate assay. MIC and sub-MIC concentrations of ZnO NPs (10–30 nm with purity greater than 99.8%) in P. aeruginosa were determined. Subsequently, biofilm formation was investigated using sub-MIC concentrations of ZnO NPs. Finally, total RNA was extracted and RT- qPCR was used to determine the expression levels of genes of mazEF, mqsRA, and higBA of TA systems.ResultsSix isolates of P. aeruginosa were found to form strong biofilms. The results showed that ZnO NPs were able to inhibit biofilm formation. In our experiments, we found that the sub-MIC concentration of ZnO NPs increased the gene expression of antitoxins mazE and mqsA and toxin higB of TA systems treated with ZnO NPs.ConclusionsIn the present study, ZnO NPs were shown to effectively inhibit biofilm formation in P. aeruginosa. Our results support the relationship between TA systems and ZnO NPs in biofilm formation in P. aeruginosa. Importantly, the expression of antitoxins mazE and mqsA was high after treatment with ZnO NPs, but not that of antitoxin higA.

Drug-tolerant persister cells in cancer: the cutting edges and future directions.

Drug-tolerant persister (DTP) cell populations were originally discovered in antibiotic-resistant bacterial biofilms. Similar populations with comparable features have since been identified among cancer cells and have been linked with treatment resistance that lacks an underlying genomic alteration. Research over the past decade has improved our understanding of the biological roles of DTP cells in cancer, although clinical knowledge of the role of these cells in treatment resistance remains limited. Nonetheless, targeting this population is anticipated to provide new treatment opportunities. In this Perspective, we aim to provide a clear definition of the DTP phenotype, discuss the underlying characteristics of these cells, their biomarkers and vulnerabilities, and encourage further research on DTP cells that might improve our understanding and enable the development of more effective anticancer therapies.