Biofilm Production Research Articles

Understanding the transitional phase of nitrate removal in three different specialty adsorbents for stormwater treatment

Excess nitrogen input from nonpoint sources driven by hydrologic and anthropogenic disturbances has become a significant ecological challenge in receiving water bodies. Moreover, nitrate removal by cost-effective filtration media has still not performed up to expectation because of the knowledge gap in the transitional phase from physicochemical to microbiological processes. This study developed three cost-effective, sustainable, and adaptable green sorption media (GSM), namely CPS (clay–perlite with sand sorption media), ZIPGEM (zero-valent iron and perlite green environmental media), and BIPGEM (biochar, zero-valent iron, and perlite-based green environmental media), made of readily available and recyclable materials, to compare nitrate removal methods for stormwater treatment and elucidate the transitional phase. The primary adsorption mechanism was chemisorption, and subsequent biofilm growth sustained removal efficiency for a long-lasting period. Adsorption kinetics, adsorption equilibrium, material characterization, and fixed-bed column experiments with a 2-stage cultivation approach were undertaken to characterize the transitional phase in these GSM, based on breakthrough curves, maximum adsorption capacity, and microbial population dynamics for nitrification and denitrification. As shown in the column study, BIPGEM had the highest adsorption capacity (7.637 mg∙g−1), followed by ZIPGEM (2.16 mg∙g−1), and CPS (0.1798mg∙g−1). The biochar in BIPGEM provides a better cultivation environment for nitrifying and denitrifying bacteria, contributing sufficient electron donors. Biofilm production and maturation enabled the BIPGEM to maintain 30 % nitrate removal in a steady state even after 2000 h of operation in the first stage. When the biofilm reached maturation, BIPGEM’s nitrate removal percentage was up to 70 % via denitrification in the second stage.

Characterization of Clonal Groups of Antibiotic-Resistant Biofilm-Forming Staphylococcus aureus Strains Isolated from Patients with Urinary Tract Infections in Isfahan, Iran

Background: Over the past decades, the role of biofilm-forming Staphylococcus aureus strains in urinary tract infections (UTIs) has garnered significant attention. Objectives: This study aimed to determine the epidemiological characteristics and diversity of S. aureus strains isolated from patients with UTIs in Isfahan, Iran, in 2017, with regard to their antimicrobial resistance, biofilm formation, and phylogenetic profiles. Additionally, the study investigated potential relationships among these factors statistically to develop efficient control and treatment approaches. Methods: All patients with symptomatic UTIs who had positive urine cultures for S. aureus during the study period at the laboratory of a referral hospital in Isfahan were included. All isolates were identified using specific primers for the nucA gene. Their biofilm formation capacity was evaluated using a combination of the microtiter plate and Congo-red agar methods. Antibiotic susceptibility testing was performed using the disk diffusion method. The presence of genes involved in biofilm formation and resistance to cefoxitin, aminoglycosides, and fluoroquinolones was detected using polymerase chain reaction (PCR). Staphylococcal cassette chromosome mec (SCCmec) typing, agr typing, and phene plate (PhP) typing were employed to investigate the diversity of collected strains. Results: Results showed that 19%, 57%, and 24% of confirmed S. aureus strains were strong, intermediate, and non-biofilm formers, respectively. The highest rate of resistance was against nalidixic acid (77%), followed by streptomycin (73%). The icaD and icaA genes had the highest frequency among biofilm-producing strains. gyrA (44%) and grlA (35%) were the most frequent genes among fluoroquinolone-resistant strains, while aph(3′)-IIIa was the most prevalent aminoglycoside-modifying enzyme gene. The majority of bacterial strains harbored SCCmec type III and agr type I. PhP typing of strains revealed the presence of 8 common types (CTs) and 14 single types (STs), with CT2 being the dominant type. Conclusions: The present investigation revealed various biofilm production capacities, antimicrobial resistance profiles, and clonal lineages in S. aureus isolated from patients with UTIs. These findings provide further insights into the epidemiology and pathogenicity of S. aureus strains in Iran, thereby improving the quality of surveillance and therapeutic protocols.

Correlation between Klebsiella Species’ Biofilm Formation and Antibiotic Resistance from Erbil Hospital Patients

The primary cause of nosocomial infections is Klebsiella pneumonia, an environmental gram-negative, encapsulated, non-motile bacterium. Antibiotic resistance and biofilm formation are two crucial characteristics that contribute to the pathogenicity of this bacterium. This research aimed to examine the relationship between antibiotic resistance and the capability of Klebsiella species isolated from Erbil hospital patients to produce biofilms. Therefore, in this study, 38 samples were collected from different laboratories and hospitals. At first, the samples were rejuvenated in Tryptic soy broth and then cultured on MacConkey agar. Gram staining and urease test were performed to identify the bacterial isolates while VITEK 2 system was performed to determine both the bacterial isolates and the antibiotic susceptibility. At last, Biofilm detection was assessed by the microliter plate technique as well as the Congo red agar method. The results revealed positive Klebsiella samples that were both resistant to some antibiotics and biofilm producers.

Lipopolysaccharide Core Truncation in Invasive Escherichia coli O157:H7 ATCC 43895 Impairs Flagella and Curli Biosynthesis and Reduces Cell Invasion Ability.

Escherichia coli O157:H7 (E. coli O157) is known for causing severe foodborne illnesses such as hemorrhagic colitis and hemolytic uremic syndrome. Although E. coli O157 is typically regarded as an extracellular pathogen and a weak biofilm producer, some E. coli O157 strains, including a clinical strain ATCC 43895, exhibit a notable ability to invade bovine crypt cells and other epithelial cells, as well as to form robust biofilm. This invasive strain persists in the bovine host significantly longer than non-invasive strains. Various surface-associated factors, including lipopolysaccharides (LPS), flagella, and other adhesins, likely contribute to this enhanced invasiveness and biofilm formation. In this study, we constructed a series of LPS-core deletion mutations (waaI, waaG, waaF, and waaC) in E. coli O157 ATCC 43895, resulting in stepwise truncations of the LPS. This approach enabled us to investigate the effects on the biosynthesis of key surface factors, such as flagella and curli, and the ability of this invasive strain to invade host cells. We confirmed the LPS structure and found that all LPS-core mutants failed to form biofilms, highlighting the crucial role of core oligosaccharides in biofilm formation. Additionally, the LPS inner-core mutants ΔwaaF and ΔwaaC lost the ability to produce flagella and curli. Furthermore, these inner-core mutants exhibited a dramatic reduction in adherence to and invasion of epithelial cells (MAC-T), showing an approximately 100-fold decrease in cell invasion compared with the outer-core mutants (waaI and waaG) and the wild type. These findings underscore the critical role of LPS-core truncation in impairing flagella and curli biosynthesis, thereby reducing the invasion capability of E. coli O157 ATCC 43895.

Downregulation of Klebsiella pneumoniae RND efflux pump genes following indole signal produced by Escherichia coli

BackgroundMore than a century has passed since it was discovered that many bacteria produce indole, but research into the actual biological roles of this molecule is just now beginning. The influence of indole on bacterial virulence was extensively investigated in indole-producing bacteria like Escherichia coli. To gain a deeper comprehension of its functional role, this study investigated how indole at concentrations of 0.5-1.0 mM found in the supernatant of Escherichia coli stationary phase culture was able to alter the virulence of non-indole-producing bacteria, such as Pseudomonas aeruginosa, Proteus mirabilis, and Klebsiella pneumoniae, which are naturally exposed to indole in mixed infections with Escherichia coli.ResultsBiofilm formation, antimicrobial susceptibility, and efflux pump activity were the three phenotypic tests that were assessed. Indole was found to influence antibiotic susceptibly of Pseudomonas aeruginosa, Proteus mirabilis and Klebsiella pneumoniae to ciprofloxacin, imipenem, ceftriaxone, ceftazidime, and amikacin through significant reduction in MIC with fold change ranged from 4 to 16. Biofilm production was partially abrogated in both 32/45 Pseudomonas aeruginosa and all eight Proteus mirabilis, while induced biofilm production was observed in 30/40 Klebsiella pneumoniae. Moreover, acrAB and oqxAB, which encode four genes responsible for resistance-nodulation-division multidrug efflux pumps in five isolates of Klebsiella pneumoniae were investigated genotypically using quantitative real-time (qRT)-PCR. This revealed that all four genes exhibited reduced expression indicated by 2^−ΔΔCT < 1 in indole-treated isolates compared to control group.ConclusionThe outcomes of qRT-PCR investigation of efflux pump expression have established a novel clear correlation of the molecular mechanism that lies beneath the influence of indole on bacterial antibiotic tolerance. This research provides novel perspectives on the various mechanisms and diverse biological functions of indole signaling and how it impacts the pathogenicity of non-indole-producing bacteria.

Phage against the Machine: The SIE-ence of Superinfection Exclusion.

Prophages can alter their bacterial hosts to prevent other phages from infecting the same cell, a mechanism known as superinfection exclusion (SIE). Such alterations are facilitated by phage interactions with critical bacterial components involved in motility, adhesion, biofilm production, conjugation, antimicrobial resistance, and immune evasion. Therefore, the impact of SIE extends beyond the immediate defense against superinfection, influencing the overall fitness and virulence of the bacteria. Evaluating the interactions between phages and their bacterial targets is critical for leading phage therapy candidates like Pseudomonas aeruginosa, a Gram-negative bacterium responsible for persistent and antibiotic-resistant opportunistic infections. However, comprehensive literature on the mechanisms underlying SIE remains scarce. Here, we provide a compilation of well-characterized and potential mechanisms employed by Pseudomonas phages to establish SIE. We hypothesize that the fitness costs imposed by SIE affect bacterial virulence, highlighting the potential role of this mechanism in the management of bacterial infections.

Curcumin loaded starch-based aerogels interfere with quorum sensing regulated virulence functions and biofilm of bacterial pathogens

Over the last two decades, the sharp escalation of antimicrobial resistance (AMR) has emerged as a formidable threat to human health. This pressing situation demands innovative interventions to combat infectious diseases. Aerogels, characterized by their nanostructured composition and high porosity, present a promising avenue. In this study, we manufactured curcumin loaded starch-based aerogels (CSA) and evaluated their impact on the quorum sensing (QS) mechanism in Gram-negative bacteria. Fourier-transform infrared (FTIR) analysis highlighted the presence of cellulose hydroxyl groups engaged in hydrogen bond formation. Thermogravimetric analysis (TGA) revealed that over 50 % of the initial mass was lost when CSA underwent heating to 350 °C. Microscopic examination showcased a uniform and compact pattern, suggesting reduced pore distribution. The elemental composition analysis indicated that carbon and oxygen constituted 23.00 % and 77.00 % of the weight, respectively. The presence of CSA resulted in over 30.4 % inhibition of violacein pigment production. Furthermore, CSA modulated pyocyanin production, pyoverdin production, LasB elastase activity, and rhamnolipid production by 34.4 %, 31.07 %, 22.7 %, and 19.7 %, respectively. The total exoproteases, cell surface hydrophobicity and exopolysaccharide production in E. coli, L. monocytogenes, S. marcescens, and P. aeruginosa, experienced a significant decrease at sub-MICs. Production of biofilms as well as the mature biofilms in test bacteria were reduced in dose-dependent manner significantly. Due to their antibiofilm and anti-quorum sensing properties, these CSAs could prove to be functional biomaterials with versatile applications, particularly in the food industry.

The correlation between signalling 2‐phenylethanol and flavour substances of sour fish and its effect on Saccharomyces cerevisiae 31

SummaryThis study aimed to investigate the interaction between 2‐phenylethanol (2‐PE) signalling molecule and volatile flavour compounds, as well as how 2‐PE would affect Saccharomyces cerevisiae 31 (Sc 31) growth and metabolism. Succession of 2‐PE in sour fish with and without Sc 31, as well as volatile flavour compounds, and its influence on cell morphology in yeast extract peptone dextrose (YPD) medium were determined. Spearman showed that the content of 2‐PE had a positive correlation with inoculated Sc 31 (r = 0.785), esters (r = 0.885), and alcohols (r = 0.758). Moreover, Sc 31 growth rate was positively impacted by 2‐PE up to 500 μm in YPD medium. This could significantly increase the production of biofilm (P &lt; 0.05) and also partially damage the cell wall and membrane. As a result, 2‐PE might affect exchange of substances between Sc 31 and the environment, which could promote the flavour generation during fermentation.

Antibiotic susceptibility and biofilm forming ability of Staphylococcus aureus isolated from Jordanian patients with diabetic foot ulcer.

Microbial biofilm is characterized by the irreversible attachment of planktonic cells to a surface and is usually associated with high antimicrobial resistance with worsening the wound healing. The objective of the study was to determine the prevalence of Staphylococcus aureus in diabetic foot ulcers (DFUs) of diabetic patients and to investigate antibiotic susceptibility patterns of these isolates. In addition to screen biofilm forming ability of isolated S. aureus. A total of 112 non-healing wound swabs of diabetic foot patients were collected and cultured on different culture media to identify and characterize 98 isolates. The S. aureus isolates were examined for their antibiotic susceptibility to different antimicrobial agents. Furthermore, S. aureus isolates were evaluated for their biofilm production capability using the Tissue Culture Plate Method (TPC). The level of icaA gene expression was determined by RT-PCR. The results of this study showed that these non-healing wounds yield positive cultures, with an average of 1.67 organisms per sample. The isolates showed highest resistance against oxacillin (95.2%) and lowest resistance against linezolid (3.7%). All isolates were biofilm producers and a significant association with the icaA gene expression level was recorded. This study showed that S. aureus isolates have a great ability to produce biofilms that are associated with the chronicity of wounds in diabetic patients. Routine screening for biofilm formers in chronic wounds and their antibiotic susceptibility testing will help in early treatment and prevent any other complications.

Metallo-β-lactamases producing Pseudomonas aeruginosa and the molecular mechanism of drug resistance variants

BackgroundMetallo-β-lactamases (MBLs) type carbapenemases are produced by pathogenic Pseudomonas spp. and exhibit carbapenemase activity. The study will look into drug resistance and the molecular mechanisms of drug-resistant Pseudomonas aeruginosa variants. MethodsA total of 74 P. aeruginosa strains were isolated from urine, pus, sputum, blood, throat swab, Foley’s catheter, and nasal swab. The isolates were screened for antibiotic susceptibility and the identified MDR strains were further tested for β-lactamase production. Multiplex PCR was used to identify the presence of mcr-1 and blaNDM-1 genes in MDR organisms. The minimum inhibitory concentration for ceftazidime and colistin was also determined, in addition to the biofilm inhibitor activity. Confocal microscopy was used to determine the production of biofilms. ResultsThe isolated P. aeruginosa strains exhibited antibiotic resistance to aminoglycosides (amikacin and gentamicin). Fifty three percent of the isolated P. aeruginosa strains produced metallo-β-lactamase and the remaining isolates were non-metallo-β-lactamase type (46.8 %) (p < 0.0001). The MIC value of β-lactamase producers against colistin ranges from 0.5 µg/mL to 6 µg/mL. The MDR bacteria exhibited mcr-1 and blaNDM−1 genes. The MDR P. aeruginosa strain treated with colistin and ceftazidime inhibited initial biofilm formation. These combination of antibiotics effectively prevented initial biofilm development than individual antibiotics (p < 0.001). ConclusionsThe current analysis detected MDR among P. aeruginosa isolates that carried drug-resistant genes.

Adaptive evolution of carbapenem-resistant hypervirulent Klebsiella pneumoniae in the urinary tract of a single patient

The emergence of carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKp) is a growing concern due to its high mortality and limited treatment options. Although hypermucoviscosity is crucial for CR-hvKp infection, the role of changes in bacterial mucoviscosity in the host colonization and persistence of CR-hvKp is not clearly defined. Herein, we observed a phenotypic switch of CR-hvKp from a hypermucoviscous to a hypomucoviscous state in a patient with scrotal abscess and urinary tract infection (UTI). This switch was attributed to decreased expression of rmpADC, the regulator of mucoid phenotype, caused by deletion of the upstream insertion sequence ISKpn26. Postswitching, the hypomucoid variant showed a 9.0-fold decrease in mice sepsis mortality, a >170.0-fold reduction in the ability to evade macrophage phagocytosis in vitro, and an 11.2- to 40.9-fold drop in growth rate in normal mouse serum. Conversely, it exhibited an increased residence time in the mouse urinary tract (21 vs. 6 d), as well as a 216.4-fold boost in adhesion to bladder epithelial cells and a 48.7% enhancement in biofilm production. Notably, the CR-hvKp mucoid switch was reproduced in an antibiotic-free mouse UTI model. The in vivo generation of hypomucoid variants was primarily associated with defective or low expression of rmpADC or capsule synthesis gene wcaJ, mediated by ISKpn26 insertion/deletion or base-pair insertion. The spontaneous hypomucoid variants also outcompeted hypermucoid bacteria in the mouse urinary tract. Collectively, the ISKpn26-associated mucoid switch in CR-hvKp signifies the antibiotic-independent host adaptive evolution, providing insights into the role of mucoid switch in the persistence of CR-hvKp.

Phylogenetic Diversity, Antibiotic Resistance, and Virulence of Escherichia coli Strains from Urinary Tract Infections in Algeria.

Urinary tract infections (UTIs) caused by Escherichia coli represent a significant public health concern due to the high virulence and antimicrobial resistance exhibited by these pathogens. This study aimed to analyze the phylogenetic diversity and antibiotic resistance profiles of Uropathogenic E. coli (UPEC) strains isolated from UTI patients in Algeria, focusing on virulence factors such as extended β-lactamase (ESBL) production, biofilm formation, and hemolytic activity. Phylogenetic grouping of 86 clinical imipenem resistant E. coli isolates showed the prevalence of group B2 (48.9%), followed by groups E (22.1%), unknown (12.8%), A (8.1%), and B1 (4.7%), and Clade I, D, Clade I, or Clade II (1.2%). The highest resistance rates were observed towards amoxicillin (86.04%), ticarcillin (82.55%), piperacillin (73.25%), nitrofurantoin (84.88%), and trimethoprim-sulfamethoxazole (51.16%). Notably, 69.8% of UPEC strains were multidrug-resistant (MDR) and 23.2% were extensively drug-resistant (XDR). Additionally, 48.9%, 42%, and 71% of strains demonstrated ESBL production, hemolytic activity, and weak biofilm production, respectively. Continuous monitoring and characterization of UPEC strains are essential to track the spread of the most resistant and virulent phylogenetic groups over time, facilitating rapid therapeutic decisions to treat infections and prevent the emergence of new resistant organisms, helping choose the most effective antibiotics and reducing treatment failure.

Prevalence of plasmid-mediated quinolone resistance genes and biofilm formation in different species of quinolone-resistant clinical Shigella isolates: a cross-sectional study

BackgroundThe purpose of this study was to look into the presence of plasmid-mediated quinolone resistance (PMQR) genes and biofilm formation in several species of clinical Shigella isolates that were resistant to quinolones.MethodsThe stool samples of 150 patients (younger than 10 years) with diarrhea were collected in this cross-sectional study (November 2020 to December 2021). After cultivation of samples on Hektoen Enteric agar and xylose lysine deoxycholate agar, standard microbiology tests, VITEK 2 system, and polymerase chain reaction (PCR) were utilized to identify Shigella isolates. The broth microdilution method was used to determine antibiotic susceptibility. PMQR genes including qnrA, qnrB, qnrC, qnrD, qnrE, qnrS, qnrVC, qepA, oqxAB, aac(6′)-Ib-cr, and crpP and biofilm formation were investigated in quinolone-resistant isolates by PCR and microtiter plate method, respectively. An enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) technique was used to determine the clonal relatedness of quinolone-resistant isolates.ResultsA total of 95 Shigella isolates including S. sonnei (53, 55.8%), S. flexneri (39, 41.1%), and S. boydii (3, 3.2%) were identified. The highest resistance rates of the isolates were against ampicillin (92.6%, n = 88/95). Overall, 42 of 95 (44.2%) isolates were simultaneously resistant against two or more quinolones including 26 (61.9%) S. sonnei and 16 (38.1%) S. flexneri. All isolates were multidrug-resistant (resistance to more than 3 antibiotics). The occurrence of PMQR genes was as follows: qnrS (52.4%), qnrA and aac(6′)-Ib-cr (33.3%), and qnrB (19.0%). The prevalence in species was as follows: 61.5% and 37.5% (qnrS), 19.2% and 56.3% (qnrA), 38.5% and 25.0 (aac(6′)-Ib-cr), and 19.2% and 18.8% (qnrB) for S. sonnei and S. flexneri, respectively. The other PMQR genes were not detected. In total, 52.8% (28/53) of quinolone-susceptible and 64.3% (27/42) of quinolone-resistant isolates were biofilm producers. Biofilm formation was not significantly different between quinolone-resistant and quinolone-susceptible isolates (P-value = 0.299). Quinolone-resistant isolates showed a high genetic diversity according to the ERIC-PCR.ConclusionIt seems that qnrS, qnrA, and aac(6′)-Ib-cr play a significant role in the quinolone resistance among Shigella isolates in our region. Also the quinolone-resistant S. flexneri and S. sonnei isolates had a high genetic diversity. Hence, antibiotic therapy needs to be routinely revised based on the surveillance findings.

Molecular and phenotypic characterization of biofilm formation and antimicrobial resistance patterns of uropathogenic staphylococcus haemolyticus isolates in Casablanca, Morocco

This study aimed to establish the correlation between antibiotic resistance and biofilm formation by Staphylococcus haemolyticus and to examine the impact of sub-inhibitory concentrations of antibiotics (sub-MICs) on biofilm formation.Antibiotic susceptibility testing was conducted using the disk diffusion method, and biofilm formation was determined using Congo red agar and microtiter plate methods. Antibiotic resistance and biofilm-associated genes were detected using polymerase chain reaction.The majority of the twenty-one S. haemolyticus isolates were multidrug-resistant, methicillin-resistant (MRSH) and biofilm producers, including 43 % of moderate biofilm producers. A significant correlation was observed between MRSH and MSSH isolates in terms of biofilm production. Vancomycin, gentamicin, and ciprofloxacin at their sub-MICs tended to promote biofilm formation. The eno gene was present in 76.2 % of strains, followed by aap, and atlE.This study revealed a strong correlation between the biofilm-forming ability and antibiotic resistance in S. haemolyticus, which underlines a crucial public health issue.

Porphyrin Production by Corynebacterium diphtheriae Strains from Clinical Isolates.

Porphyrins are intermediate metabolites involved in the biosynthesis of vital molecules, including heme, cobalamin, and chlorophyll. Bacterial porphyrins are known to be proinflammatory and are associated with biofilm formation. This study investigated porphyrin production by strains of Corynebacterium diphtheriae using emission spectroscopy, high-performance liquid chromatography with fluorescence detection, diode array detector, and mass spectrometry. Emission spectroscopy revealed characteristic porphyrin emission spectra in all strains, with coproporphyrin III predominating. Qualitative analysis by different chromatographic methods identified coproporphyrin III, uroporphyrin I, and protoporphyrin IX in all strains. Quantitative analysis revealed strain-dependent coproporphyrin III production. Further studies are required to investigate the relationship between porphyrin production and the virulence potential of Corynebacterium diphtheriae.

Agroclimatic zone-wise prevalence of biofilm forming methicillin resistant staphylococcus aureus in West Bengal

The present study envisages the prevalence of biofilm producing, methicillin resistant Staphylococcus aureus, isolated from cattle milk of different agro-climatic zones of an eastern state of India, West Bengal. The milk samples (n=168) were collected from three predominant agro- climatic zones of South Bengal, viz. new alluvial, coastal saline and red laterite. A total of 146 Staphylococcus aureus isolates were recovered, confirmed by biochemical tests and polymerase chain reaction (PCR). Through PCR of mecA gene, it was found that 45 strains were methicillin resistant, encompassing 28.57% new alluvial zone, 30.19% red laterite region and 29.16% coastal saline region. Determination of biofilm formation was done, both phenotypically by microtiter plate method and genotypically by PCR of icaA and icaD genes, specific for biofilm formation in Staphylococcus aureus. Phenotypic study revealed that 14 isolates were strong biofilm producers, 79 were moderate and the 47 were weak biofilm producers. PCR studies also showed that a total of 30 isolates were icaA positive and 25 isolates were icaD positive. Although no significant difference was found in the occurrence of icaA positive isolates, the possession of icaD by the Staphylococcus aureus isolates varied significantly according to different agro-climatic zones.

Lacticaseibacillus rhamnosus inhibits the development of dental caries in rat caries model and in vitro

ObjectivesDental caries result from a microbial imbalance in the oral cavity. Probiotics ecologically modulate the oral microflora to prevent caries. This study evaluated the anti-cariogenic effects of two Lacticaseibacillus rhamnosus strains in vitro and in vivo to provide a more theoretical basis for its clinical applications in caries prevention. MethodsIn the study, cariogenic biofilms were grown with L. rhamnosus (LGG) or L. rhamnosus ATCC 7469 and analyzed. Quantitative real-time PCR (qPCR), Scanning Electron Microscope (SEM), and Confocal laser scanning microscope (CLSM) were used to detect the changes in the composition and architectures; cariogenic activity was measured by the lactic acid production and Transverse Microradiography (TMR). The effects of LGG on the 12 Sprague-Dawley rat caries model were assessed using Keyes scores and micro-CT analysis. Oral microbiome changes were evaluated through 16S rRNA gene high-throughput sequencing. ResultsL. rhamnosus can reduce cariogenic bacteria in biofilm by 14.7 % to 48.9 %, with LGG exhibiting more potent inhibitory effects. Both strains of L. rhamnosus can adhere to the surface of biofilms, reduce the extracellular polysaccharides (EPS) matrix, and loosen the biofilm structure. L. rhamnosus inhibited cariogenic activity by reducing the lactic acid production in biofilms. The bovine enamel blocks presented lower mineral loss values and lesion depth values in the group Core+L.rh and Core+LGG. LGG-ingested rats had significantly lower levels of moderate dentin lesions and higher mineral density than the control group. The 16 s rRNA gene sequencing revealed that LGG regulated the beta diversity of the oral microbial community in the rat dental caries model. ConclusionsThis study revealed the promising potential of L. rhamnosus, especially the LGG strain, in the ecological prevention of dental caries. Clinical SignificanceProbiotics may provide a strategy for preventing caries by regulating the oral microecological balance. The study revealed the promising anti-caries potential of the LGG probiotic strain in vivo and in vitro. It is expected that LGG could be used as an oral probiotic for the clinical prevention and treatment of caries.

Characterization of biofilm-forming ability and antibiotic resistance profiles of clinical Pseudomonas aeruginosa isolates from patients with cystic fibrosis

Background: Pseudomonas aeruginosa is a key pathogen in cystic fibrosis (CF) associated infections, known for its biofilm-forming ability and resistance to multiple antibiotics. Understanding the relationship between biofilm production and antibiotic resistance profiles in clinical isolates can guide treatment strategies. Aims and Objectives: The aims of this study were to investigate the biofilm-forming abilities and antibiotic resistance profiles of clinical P. aeruginosa isolates from patients with CF and to determine the relationship between these two factors and their impact on treatment efficacy. Materials and Methods: A cross-sectional study was conducted on 100 clinical isolates of P. aeruginosa from CF patients. Biofilm-forming capacity was categorized as strong, moderate, or weak based on quantitative assays. Antibiotic resistance was assessed for ciprofloxacin, tobramycin, ceftazidime, meropenem, and piperacillin/tazobactam. Multi-drug resistance was defined as resistance to three or more antibiotic classes. Statistical correlations between biofilm-forming capacity and resistance levels were evaluated using the Chi-square tests. Results: Fifty-six percentages of isolates were strong biofilm producers, while 34% and 10% were moderate and weak producers, respectively. The highest antibiotic resistance was observed against ciprofloxacin (60%), followed by tobramycin (50%) and ceftazidime (45%). Forty percentages of the isolates were classified as multi-drug resistant. Strong biofilm producers demonstrated a significantly high correlation with antibiotic resistance (P&lt;0.05). Two predominant clonal groups were identified among the isolates, suggesting a possible clonal spread of resistance traits. Conclusion: The study confirms a strong association between robust biofilm production and heightened antibiotic resistance in P. aeruginosa isolates from CF patients. These findings highlight the need for targeted therapeutic strategies to disrupt biofilm formation and curb resistance spread.

Two cinnamoyl hydroxamates as potential quorum sensing inhibitors against Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a ubiquitous pathogen that causes various infectious diseases through the regulation of quorum sensing (QS). The strategy of interfering with the QS systems of P. aeruginosa, coupled with a reduction in the dosage of conventional antibiotics, presents a potential solution to treating infection and mitigating antibiotic resistance. In this study, seven cinnamoyl hydroxamates were synthesized to evaluate their inhibitory effects on QS of P. aeruginosa. Among these cinnamic acid derivatives, we found cinnamoyl hydroxamic acid (CHA) and 3-methoxy-cinnamoyl hydroxamic acid (MCHA) were the two most effective candidates. Furtherly, the effect of CHA and MCHA on the production of virulence factors and biofilm of P. aeruginosa were evaluated. Ultimately, our study may offer promising potential for treating P. aeruginosa infections and reducing its virulence. The disc diffusion test were conducted to evaluate inhibitory effects on QS of seven cinnamoyl hydroxamates. The influence of CHA and MCHA on the production of virulence and flagellar motility of P. aeruginosa was furtherly explored. Scanning electron microscopy (SEM) experiment were conducted to evaluate the suppression of CHA and MCHA on the formed biofilm of P. aeruginosa. RT-qPCR was used to detect rhlI, lasA, lasB, rhlA, rhlB, and oprL genes in P. aeruginosa. In silico docking study was performed to explore the molecular mechanism of CHA and MCHA. The synergistic effects of CHA with gentamicin were detected on biofilm cell dispersal. After treatment of CHA or MCHA, the production of multiple virulence factors, including pyocyanin, proteases, rhamnolipid, and siderophore, and swimming and swarming motilities in P. aeruginosa were inhibited significantly. And our results showed CHA and MCHA could eliminate the formed biofilm of P. aeruginosa. RT-qPCR revealed that CHA and MCHA inhibited the expression of QS related genes in P. aeruginosa. Molecular docking indicated that CHA and MCHA primarily inhibited the RhlI/R system in P. aeruginosa by competing with the cognate signaling molecule C4-HSL.Additionally, CHA exhibited potent synergistic effects with gentamicin on biofilm cell dispersal. P. aeruginosa is one of the most clinically and epidemiologically important bacteria and a primary cause of catheter-related urinary tract infections and ventilator-associated pneumonia. This study aims to explore whether cinnamoyl hydroxamates have inhibitory effects on QS. And our results indicate that CHA and MCHA, as two novel QSIs, offer promising potential for treating P. aeruginosa infections and reducing its virulence.

Biofilm formation, agr typing and antibiotic resistance pattern in methicillin-resistant Staphylococcus aureus isolated from hospital environments.

Biofilm development significantly enhances the virulence of methicillin-resistant Staphylococcus aureus (MRSA), leading to severe infections and decreased susceptibility to antibiotics, especially in strains associated with hospital environments. This study examined the occurrence of MRSA, their ability to form biofilms, agr typing, and the antibiotic resistance profiles of biofilm-forming MRSA strains isolated from environmental surfaces at Mymensingh Medical College Hospital (MMCH). From 120 swab samples, 86 (71.67%) tested positive for S. aureus. MRSA was identified in 86 isolates using the disk diffusion technique, and by polymerase chain reaction (PCR), 56 (65.1%) isolates were confirmed to carry the mecA gene. The Crystal Violet Microtiter Plate (CVMP) test revealed that 80.35% (45 isolates) were biofilm-forming and 19.6% (11 isolates) were non-biofilm-forming. Out of 45 biofilm producer isolates 37.5% and 42.9% isolates exhibited strong and intermediate biofilm-forming characteristics, respectively. Molecular analysis revealed that 17.78% of MRSA isolates carried at least one gene related to biofilm formation, specifically icaA, icaB, and icaD genes were discovered in 13.33%, 8.89%, 6.67% of the MRSA isolates, respectively. In agr typing, the most prevalent group was agr I (71.11%), followed by group III (17.78%) and group II (11.11%). Group IV was not detected. The distribution of agr gene groups showed a significant difference among biofilm-forming isolates (p < 0.05). In agr group I, 18.75% of isolates carried the icaA gene, 12.5% carried the icaB gene, and 9.37% carried the icaD gene. Biofilm-forming genes were not detected in any of the isolates from agr groups II or III. There are no statistically significant differences between agr groups and the presence of these genes (p > 0.05). Antibiotic resistance varied significantly among agr groups, with agr group I displaying the highest resistance, agr group II, and agr group III exhibiting the least resistance (p < 0.05). Seventy-three (73.3%) of the isolates were multi-drug resistant, with agr group I displaying nineteen MDR patterns. The occurrence of MRSA in hospital environments and their capacity to form biofilm raises concerns for public health. These findings support the importance of further research focused on agr quorum sensing systems as a basis for developing novel antibacterial agents.