Moderate Biofilm Producers Research Articles

Evaluation of biofilm formation and carbapenem resistance in Klebsiella pneumoniae isolated from clinical samples at a rural hospital in western Uttar Pradesh

ABSTRACT Introduction: Klebsiella pneumoniae commonly causes healthcare-associated infections and shows multidrug resistance. K. pneumoniae can produce biofilm. Carbapenem resistance in K. pneumoniae is due to the production of carbapenemases mainly. This study was done to evaluate the formation of biofilm and carbapenemase resistance in K. pneumoniae isolates. Material and Methods: A total of 110 K. pneumoniae isolated from various clinical samples were taken, the antibiotic susceptibility test was done by the Kirby disk diffusion method, and biofilm detection was done by the tissue culture plate method. All the carbapenem-resistant isolates were confirmed by multiplex real-time PCR (mPCR). Those found positive for any of the carbapenemase genes were tested by the modified Hodge test (MHT), modified carbapenem inactivation method (mCIM), and ethylenediamine tetraacetic acid (EDTA)-modified carbapenem inactivation method (eCIM). Results: Out of 110 isolates, 66% (72/110) were carbapenem-resistant (suggestive of carbapenemase producers) by Kirby-Bauer disk diffusion but 58% (42/72) of Klebsiella isolates were confirmed for carbapenemase production by mPCR. Maximum number of carbapenemase gene were New Delhi metallo-β-lactamase (NDM) 52% (N = 22), 29% (N = 12) coproducers (NDM+OXA-48), and lowest in oxacillinase (OXA-48), 19% (N = 8). The overall sensitivity of MHT and mCIM+eCIM was 62% and 93%, and specificity was 88% and 97%, respectively. Our study showed that moderate biofilm producers were 51% (N = 56) K. pneumoniae isolates, strong biofilm producers 27% (N = 30), and 22% (N = 30) were weak/non-biofilm producers. We also found the correlation between biofilm formation and carbapenem-resistant K. pneumoniae (CR-KP) genes was statistically significant with a P value of 0.01*<0.05. Conclusion: Most isolates of K. pneumoniae demonstrated a wide range of antibiotic resistance and were biofilm producers. Our results indicated that the combination of mCIM with eCIM showed high sensitivity and specificity to detect CR-KP compared with MHT.

Antibiotic Resistance, Biofilm Formation, and Identification of FimH and FimA Adhesion Genes in Uropathogenic Escherichia Coli (UPEC) Isolated from Patients in Baghdad Province

Urinary tract infections (UTIs) are one of the most common infectious disorders worldwide. The most frequent cause of UTIs is uropathogenic Escherichia coli (UPEC). The current study is designed to assess the resistance of antibiotics, the formation of biofilm, and the detection of adhesion genes in E. coli isolated from patients with UTIs. A hundred and fifty samples were collected from patients with confirmed UTIs in Baghdad province during the period of October 2022 to February 2023. Isolation and identification of E. coli were performed using cultural characteristics, gram staining, and biochemical examination, and the results indicated that 52 (35%) isolates were identified as E. coli. Eleven antibiotic discs were utilized to estimate the sensitivity of E. coli isolates. 98% of isolates were resistant to ampicillin, followed by 75.5% of isolates resistant to both trimethoprim/sulfamethoxazole and fosfomycin, followed by 73.5, 71.4, 67.3, 53.1, and 6.1% of isolates resistant to imipenem, ceftazidime, nitrofurantoin, cefepime, and amikacin. All isolates were sensitive to piperacillin-tazobactam, ciprofloxacin, and aztreonam at 100%, 100%, and 98%, respectively. The microtiter plate method was utilized to estimate the ability of E. coli to form biofilm. The majority of isolates (69.2%; n = 36) were moderate biofilm producers, while 19.2% (n = 10) and 11.5% (n = 6) of isolates were strong and weak biofilm producers, respectively. Polymerase Chain Reactions (PCR) were applied to determine the gene expression level of adhesion genes (fimH and fimA genes). Forty isolates harbored both fimH and fimA genes.

Antifungal susceptibility and biofilm production of Candida species- causative agents of female genital tract infections.

Recurrent vulvovaginal candidosis (RVVC) is a chronic infection affecting 8-10% of women worldwide. Biofilm production of the infecting species and reduced sensitivity to antimycotics could contribute to the recurrence of this infection. This study aimed to examine the biofilm production ability and antifungal susceptibility of genital yeast isolates to determine their virulence potential. Matrix-assisted laser desorption in ionization-time of flight mass spectrometry (MALDI-TOF MS) was used to identify 300 Candida species. Using crystal violet method, strains were categorized into non-producers, weak, moderate, and strong biofilm producers (BFP). Antifungal susceptibility testing was performed using commercial Integral System YEASTS Plus test (ISYPT) and broth microdilution method (BMM). MALDI-TOF MS identified 150 Candida albicans, 124 non-albicans Candida (NAC), and 26 Saccharomyces cerevisiae strains. Within 138 (46.0%) BFP, 23 (16.7%) were strong, 44 (31.9%) moderate, and 71 (51.4%) weak. BMM was done for 43 BFP selected isolates with nystatin MIC ˃1.25 μl, fluconazole MIC ˃64 μl, and clotrimazole MIC ˃1.0 μl determined by ISYPT. Compared to all examined isolates, BMM confirmed that: i) C. albicans and NAC BFP showed low sensitivity to fluconazole (12% and 4%, respectively); ii) all BFP showed low sensitivity to nystatin (12.7% C. albicans, 14.5% NAC, and 23.1% S. cerevisiae); iii) clotrimazole in vitro was the most efficient regarding C. albicans and S. cerevisiae strains, but in 4.0% NAC BFP for this antimycotic higher MIC was established. Novel antimycotics or possible combinations of antifungal agents and natural products could be a new treatment option for RVVC.

Biofilm Production Activity and Antibiotics Susceptibility of Streptococcus mutans Isolates from Patients with Dental Caries

Dental caries, a prevalent chronic infectious ailment, commences with the adherence of cariogenic bacteria to the teeth, particularly Streptococcus mutans. These microorganisms utilize sugars to produce acid, resulting in the gradual demineralization of the dental structure. Streptococcus mutans a gram positive bacterium is found to play the most significant role in the dental caries in terms of being the earliest colonizer in the formation of caries. The current research focuses on the isolation and identification of the Streptococcus mutans pathogen in individuals affected by dental caries. The isolated strains were cultured on specific media, with one hundred samples of dental caries obtained from human teeth. Among these samples, fifty were identified as positive bacterial isolates utilizing MS-agar (Mitist Salivares agar), with ten isolates categorized under the group Streptococci and twenty isolates identified as S. mutans. Identification of bacterial isolates was carried out based on morphological and cultural characteristics, biochemical tests, and VITEK-II. Additionally, an antibiogram assay was conducted to assess the susceptibility and resistance of the pathogen to various drugs. The findings revealed that all locally isolated S. mutans strains displayed 100% resistance to Cefepime, while showing different levels of resistance and susceptibility to other antibiotics such as Doxycycline, Amoxicillin-clavulanic acid, Trimethoprim-sulfamethoxazole, Vancomycin, Tetracycline, Ampicillin, and Meropenem. In this study, ELISA was employed to differentiate between Streptococcus. mutans isolates that produced biofilms and those that did not by utilizing the optical density at 630 nm. Bacterial biofilms contribute to chronic diseases that pose challenges in treatment. Twenty distinct isolates were evaluated for their biofilm-forming capabilities, with 9 (45%) identified as strong biofilm producers, 7 (35%) classified as moderate biofilm producers, and 4 (20%) categorized as weak biofilm producers

Multidrug Resistant Group B Streptococcus Isolates from Pregnant Women in Delta State, Nigeria.

&lt;b&gt;Background and Objective:&lt;/b&gt; Group B &lt;i&gt;Streptococci&lt;/i&gt; (GBS) are globally recognized as a major risk factor for neonatal infections and various obstetric complications. More so, biofilm formation has been suggested to be important for GBS pathogenesis. The aim of this study was to determine the prevalence and antibiotic susceptibility pattern of GBS among pregnant women and their capacity to form biofilm. &lt;b&gt;Materials and Methods:&lt;/b&gt; A total of 87 pregnant women at 34 to 37 weeks' gestation aged 17-45 years were recruited from 3 healthcare centres in Delta State, Nigeria. Cultures for the isolation of GBS were carried out using recto-vaginal swabs, according to standard microbiological methods. All strains isolated were used for susceptibility tests to various antibiotics as recommended by CLSI using the disk-diffusion method. &lt;b&gt;Results:&lt;/b&gt; The overall prevalence of GBS colonization among pregnant women was 43.6% (38/87). The &lt;u&gt;<&lt;/u&gt;30 age group had the highest rate of GBS colonization. Resistance to erythromycin and vancomycin was 48.2 and 66.4%, respectively. The fluoroquinolones had the lowest resistant rates with no isolate showing resistance to ofloxacin. Multidrug resistance (MDR) (&lt;u&gt;>&lt;/u&gt;3 drug classes) was detected in 73.7% (28/38) of the GBS isolates. All GBS isolated in this study were either strong, moderate or weak biofilm producers. However, most 28 (73.7%) were strong biofilm producers. Resistance of GBS isolates to erythromycin and vancomycin, drugs used for treating GBS infection was high. &lt;b&gt;Conclusion:&lt;/b&gt; This suggested the importance of testing antimicrobial susceptibilities in GBS colonized pregnant women in order to guide antibiotic therapy and minimize newborn infection and co-morbidity.

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.

Evaluation of biofilm formation by bacteria isolated from engine oil-contaminated soil and exploring its bioremediation potential in vitro

This study investigated the biofilm-forming ability of bacteria isolated from engine oil-contaminated soil and assessed their potential for bioremediation. Fifteen engine oil contaminated soil samples were collected and inoculated into Bushnell-Haas (BH) broth supplemented with 2 % engine oil. After incubation, bacterial colonies were isolated and identified using 16s RNA sequencing. The tube adherence test also known as Christensen method and modified microtiter plate quantification of biofilm formation by Stepanović et al. (2000) [1], was used to study bacterial biofilm as tube test and the microtiter-plate test are the most frequently used techniques for quantifying biofilm formation was used to study bacterial biofilm. Both quantitative (OD measurements) and qualitative (visual observation) methods were employed, classifying the six isolates into three categories: strong, moderate, and low biofilm producers. Among the 6 bacterial isolates, Stutzerimonas stutzeri exhibited the highest biofilm-forming ability.Oil degradation capacity of the isolates were tested individually as monocultures and co-cultures with other isolates. Results showed that the co-culture of Stutzerimonas stutzeri and Klebsiella quasipneumoniae showed significantly higher oil degradation compared to their monoculture and other co-cultures, suggesting synergistic interactions between the two bacterial species.However, Stutzerimonas stutzeri alone exhibited good oil degradation potential and is more useful for bioremediation but Klebsiella quasipneumoniae's rare pathogenicity, may not be suitable for field application. This study highlights biofilm-forming bacteria, particularly Stutzerimonas stutzeri, as a potential solution for bioremediating engine oil-contaminated sites.

Biofilm-producing ability of methicillin-resistant Staphylococcus aureus clinically isolated in China

BackgroundStaphylococcus aureus, a commensal bacterium, colonizes the skin and mucous membranes of approximately 30% of the human population. Apart from conventional resistance mechanisms, one of the pathogenic features of S. aureus is its ability to survive in a biofilm state on both biotic and abiotic surfaces. Due to this characteristic, S. aureus is a major cause of human infections, with Methicillin-Resistant Staphylococcus aureus (MRSA) being a significant contributor to both community-acquired and hospital-acquired infections.ResultsAnalyzing non-repetitive clinical isolates of MRSA collected from seven provinces and cities in China between 2014 and 2020, it was observed that 53.2% of the MRSA isolates exhibited varying degrees of ability to produce biofilm. The biofilm positivity rate was notably high in MRSA isolates from Guangdong, Jiangxi, and Hubei. The predominant MRSA strains collected in this study were of sequence types ST59, ST5, and ST239, with the biofilm-producing capability mainly distributed among moderate and weak biofilm producers within these ST types. Notably, certain sequence types, such as ST88, exhibited a high prevalence of strong biofilm-producing strains. The study found that SCCmec IV was the predominant type among biofilm-positive MRSA, followed by SCCmec II. Comparing strains with weak and strong biofilm production capabilities, the positive rates of the sdrD and sdrE were higher in strong biofilm producers. The genetic determinants ebp, icaA, icaB, icaC, icaD, icaR, and sdrE were associated with strong biofilm production in MRSA. Additionally, biofilm-negative MRSA isolates showed higher sensitivity rates to cefalotin (94.8%), daptomycin (94.5%), mupirocin (86.5%), teicoplanin (94.5%), fusidic acid (81.0%), and dalbavancin (94.5%) compared to biofilm-positive MRSA isolates. The biofilm positivity rate was consistently above 50% in all collected specimen types.ConclusionsMRSA strains with biofilm production capability warrant increased vigilance.

Anti-biofilm Activity of Phage ΦKAB and Colistin Against Carbapenem Resistance &lt;i&gt;Acinetobacter baumannii&lt;/i&gt;

Background: Carbapenem-resistant A. baumannii (CRAB) bacterium is difficult to treat with available antimicrobial agents leading to use a few antibiotics such as colistin as an option for treatment. However, the use of the colistin has serious side effects and leads to developing bacterial resistance. Objective: This study aimed to determine effectiveness of phage/colistin combination to inhibit biofilm formation of CRAB. Methods: Sixty clinical A. baummanii isolates were identified with the VITEKII system and 16S rRNA gene. The antibiotic susceptibility and detection of Oxacillinases genes were tested for all isolates. Results: The antibiotic sensitivity of A. baumannii isolates showed a high resistance percentage to Ceftriaxone (CRO) with 92% (55 isolates), and Cefotiam (CTF) with 87% (52 isolates), while the lowest percent related to Colistin (CO) with 17% (10 isolates). The results of antibiotic resistance and Oxacillinases genes reported that only 14 isolates from the current study were CRAB. Phage and host A. baumannii of phage were isolated and characterized previously. Biofilm production assay of CRAB isolates were showed that among 14 isolates including the phage host: 57% (8) was weak and 43% (6) was moderate biofilm producer. Therefore, the synergistic effect of a combination ΦKAB phage in MOI (10) and Colistin with MIC=8 µg/ml against CRAB isolates was evaluated and showed complete transfer of isolates by 100 % to weak biofilm producer compared with CRAB isolates. Conclusions: Among 60 isolates of A. baumannii, most isolates were MDR 44 (73.3%) and only 14 (23.3%) isolates were CRAB that were 57% weak and 43% moderate biofilm producers. The mixture of Colistin/ΦKAB phage could inhibit and reduce biofilm forming in CRAB isolates. Overall, the present study may provide evidence on the capability of the isolated phage to serve as a novel strategy to treat infections caused by MDR A. baumannii.

Coexistence of β-lactamase genes and biofilm forming potential among carbapenem-resistant Acinetobacter baumannii in Lahore, Pakistan.

Our goal was to investigate the antimicrobial resistance due to beta-lactamase genes and virulent determinants (biofilm-forming ability) expressed by Acinetobacter collected from health settings in Pakistan. A cross-sectional study was conducted for the molecular characterization of carbapenemases and biofilm-producing strains of Acinetobacter spp. Two twenty-three imipenem-resistant Acinetobacter isolates were analyzed from 2020 to 2023.The combination disk test and modified hodge test were performed. Biofilm forming ability was determined by polystyrene tube assay. Multiplex polymerase chain reaction (PCR) for virulent and biofilm-forming genes, and 16S rRNA sequencing were performed. 118 (52.9%) carbapenem-resistant Acinetobacter (CR-AB) were isolated from wounds and pus, 121 (54.2%) from males, and 92 (41.2%) from 26-50-years-olds. More than 80% of strains produced β-lactamases and carbapenemases. Based on the PCR amplification of the ITS gene, 174 (78.0%) CR-AB strains were identified from CR-Acinetobacter non-baumannii (ANB). Most CR-AB were strong and moderate biofilm producers. Genetic analysis revealed the blaOXA-23, blaTEM, blaCTX-M blaNDM-1 and blaVIM were prevalent in CR-AB with frequencies 91 (94.8%), 68 (70.8%), 19 (19.7%), 53 (55.2%), 2 (2.0%) respectively. Among virulence genes, OmpA was dominant in CR-AB isolates from wound (83, 86.4%), csuE 63 (80.7%) from non-wound specimens and significantly correlated with blaNDM and blaOXA genes. Phylogenetic analysis revealed three different clades for strains based on specimens. CR-AB was highly prevalent in Pakistan and associated with wound infections. The genes, blaOXA-23, blaTEM, blaCTX-M, and blaNDM-1 were detected in CR-AB. Most CR-AB were strong biofilm producers with virulent genes OmpA and csuE.

Identification and characterization of microorganisms isolated from noncompliant or atypical dairy products in Canada

Despite good manufacturing practices and rigorous cleaning and sanitizing procedures established in dairy processing plants, microbiological contamination remains the main cause of products being non-compliant and/or atypical and hence not fit for human consumption. The objective of this study was to isolate, identify and characterize bacteria, yeasts and molds associated with substandard dairy products in Canada and to create a collection of reference isolates. In addition to conventional microbiological characterization, each isolate was tested for biofilm-forming ability and susceptibility to heat, antimicrobial agents, and common industrial disinfectants. Among the 105 microbial strains isolated from pasteurized milk, cream, and cheese samples, 24 bacterial isolates, belonging mainly to the genus Pseudomonas, were shown to be moderate or strong biofilm producers in 96-well plates and highly resistant to peracetic acid (100 ppm, 5 min contact time) and sodium hypochlorite (70 ppm, 5 min contact time). In addition, 56 bacterial isolates, including Acinetobacter baumannii, Enterobacter bugandensis, Klebsiella pneumoniae and Pseudomonas spp., were found resistant to ampicillin, fosfomycin and/or ceftriaxone, while 14 others, such as Bacillus spp. and Macrococcus spp., withstood a heat treatment equivalent to low-temperature long-time pasteurization (63°C for 30 min). This descriptive study provides new information on potential problematic microorganisms in dairies and will guide the development of novel control strategies intended to prevent and reduce microbiological contamination and the associated economic losses.

Fusarioid keratitis and other superficial infections: A 10-years prospective study from Northeastern Brazil.

Fusarium and allied genera (fusarioid) species are common colonizers of roots and aerial plant parts, or act as phytopathogens in forestry and horticultural or grain crops. However, they can also cause a wide range of infections in humans, including onychomycosis, cutaneous and invasive infections. Fusarioid keratitis is characterized by an infection of the cornea with a suppurative and ulcerative appearance, which may cause damage to vision and permanent blindness. The aim of the present study was to investigate the prevalence of fusarioid species, biofilm formation and antifungal susceptibility profiling of clinical isolates recovered from patients with keratitis and dermatomycoses. The study was performed between March, 2012-December, 2022. Demographic, clinical and epidemiological data of patients were also collected. In the present study, most of the patients with keratitis were male (74%), had a median age of 42 years old, worked with plant material or debris and 26% of them reported eye trauma. Regarding dermatomycosis, most of patients were female and exhibited toenail lesions. Forty-seven isolates belonged to the genus Neocosmospora (78.33%), nine to the Fusarium fujikuroi (15%) and four to the Fusarium oxysporum (6.66%) species complexes. Several strains were moderate biofilm producers, specifically among Fusarium annulatum. Most strains showed increased MICs to amphotericin B and ketoconazole and low MICs to itraconazole. MICs ranged from 0.25 to 16 μg/mL for amphotericin B, 0.0625 to >16 μg/mL for ketoconazole and 0.125 to 8 for itraconazole. It is possible to conclude that fusarioid keratitis in Northeastern Brazil is an important and neglected disease, given the high number of cases, increased need for keratoplasty and poor outcome of the disease.

Assessment of antimicrobial resistance, biofilm formation, and surface modification potential in hospital strains of Pseudomonas aeruginosa and Klebsiella pneumoniae

The occurrence of healthcare-associated infections is a multifactorial phenomenon related to hospital space contamination by bacteria. The ESKAPE group, specifically Pseudomonas aeruginosa and Klebsiella pneumoniae, play a relevant role in the occurrence of these infections. Therefore, comprehensive research is needed to identify characteristics that justify the prevalence of these species in the healthcare environment. In this line, the study aimed to determine the antimicrobial resistance, biofilm formation, and the potential for polymer degradation in a collection of 33 P. aeruginosa strains and 2 K. pneumoniae strains sampled from various equipment and non-critical surfaces in a Portuguese hospital. Antimicrobial susceptibility tests revealed that none of the strains was categorized as multidrug-resistant (non-MDR). An assessment of their biofilm-forming capabilities indicated that 97 % of the strains exhibited biofilm-producing characteristics. Notably, within this group, the majority of P. aeruginosa and half of K. pneumoniae strains were classified as strong biofilm producers. Furthermore, the strains were evaluated for their potential to cause damage or change medical devices, namely infusion sets, nasal cannula, and urinary catheters. Three P. aeruginosa strains, two strong and one moderate biofilm producers, showed the highest ability to modify surfaces of the nasal cannula and infusion sets. Additionally, the Chi-square test revealed a statistically significant relationship between the presence of P. aeruginosa strains and the water accession spots. In conclusion, this work suggests that bacteria from this group hold a significant ability to grow in the healthcare environment through the degradation of non-critical materials. This suggests a potential concern for the persistence and proliferation of these organisms in hospital environments, emphasizing the importance of robust infection control measures to mitigate the risks associated with bacterial growth on such surfaces.

Probiotic Effect of Streptococcus dentisani on Oral Pathogens: An In Vitro Study.

Probiotics, including Streptococcus dentisani, have been proposed as an alternative to re-establish the ecology of the oral cavity and inhibit the formation of pathogenic biofilms. The main objective of this work was to assess the probiotic ability of S. dentisani against Streptococcus mutans, Streptococcus mitis, and Candida albicans biofilms. The ability of the strains to form a monospecies biofilm and the probiotic potential of S. dentisani using the competition, exclusion, and displacement strategies were determined. All strains were moderate biofilm producers. The ability of S. dentisani to compete with and exclude S. mutans and S. mitis during biofilm formation was not significant. However, S. dentisani significantly reduced pathologic streptococcal biofilms using the displacement strategy. Also S. dentisani reduced the formation of the C. albicans biofilm mainly through competition and displacement. In vitro, S. dentisani exhibited probiotic potential to reduce the formation of potentially pathogenic biofilms. Further investigation is required to understand the biofilm-inhibiting mechanisms exhibited by this probiotic strain.

Emerging threats of high biofilm formation and antibiotic resistance in clinical methicillin-resistant Staphylococcus aureus (MRSA) isolates from Pakistan

ObjectivesThis multicenter study, conducted from a One Health perspective, aimed to comprehensively examine the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) infections and their biofilm-forming capabilities in Pakistan. Phylogenetic analysis of the study isolates was also performed. MethodsA total of 150 MRSA isolates were screened from various clinical samples using Cefoxitin antibiotic discs. Genotypic confirmation was conducted through mecA, S. aureus-specific nuc, and 16S rRNA genes. Biofilm formation was assessed using Congo red agar (CRA) and slime layer quantification methods. The intercellular adhesion (ica) operon genes, specifically icaA and icaD, were detected. Phylogenetic analysis utilized the 16S rRNA sequences. Statistical associations between various parameters were determined using chi-square analysis. ResultsThe presence of the mecA gene was observed in 131 out of 150 isolates (87.3%). CRA identified 28% and 40% of isolates as strong and moderate biofilm producers, respectively, while 9.3% were classified as non-biofilm producers. The slime layer assay exhibited higher sensitivity, classifying only 4.7% of isolates as non-biofilm producers. Biofilm-forming genes icaA and icaD were detected in 85.3% and 86.7% of the isolates, respectively. Antibiotic resistance was more prevalent among biofilm-forming isolates, particularly against ciprofloxacin, levofloxacin, erythromycin, trimethoprim-sulfamethoxazole, and fosfomycin. Ceftaroline demonstrated efficacy irrespective of biofilm-forming abilities. Conversely, non-biofilm producers exhibited complete susceptibility to clarithromycin and tigecycline. ConclusionsClinical MRSA strains exhibit a substantial potential for biofilm formation, contributing to a resistant phenotype. Routine antibiotic testing in clinical settings that overlook the biofilm aspect may lead to the failure of empiric antibiotic therapy.

Effect of Piper Cubeba Extract on Biofim Producing Pseudomonas Aeruginosa Isolated from Clinical Samples

Background: The species of Pseudomonas aeruginosa is a nosocomial bacterium that increases the risk of multidrug-resistant infections, especially in vulnerable patients. The plant Piper cubeba, commonly referred to as cubeb or tailed pepper, it is specifically cultivated for its fruit and essential oil. The plant belongs to the genus Piper. The study aims at assessing the effectiveness of Piper cubeba extract against different P. aeruginosa strains that could produce biofilm and carry blaIMP gene. Method: Isolates from 85 samples (17 sputum, 40 urine and 28 diarrhea samples). Collected from Al Nasiriyah Teaching Hospital during the period of December 2021 to March 2022. Samples were cultivated using routine cultivation methods, catalase test, oxidase reaction, and the Api20E system were performed to identify P. aeruginosa. Results: Out of a total of 17 isolated samples of Pseudomonas aeruginosa, 3 (17.64%) were from sputum, 6(35.29%) from urine, and 8 (47.05%) diarrhea samples. Out of 17 isolates 6 were sensitive in stool isolates 4 isolates were moderate sensitive and 7 isolates were resistance. Conclusion at concentrations of 25, 50, 75, and 100 mg/mL for Piper cubeba, respectively, the greatest MBC of plant extracts was noted. Also, all the isolates were capable of producing a biofilm by using presterilized 96-well polystyrene microtiter plates. Out of 17 isolates, 9 had a potent biofilm ability and the remaining 7 were moderate biofilm producers. Additionally, 7 of the isolates (41.17%) carried the blaIMP gene. Conclusion: Piper cubeba extract has potential as an antimicrobial agent against both sensitive and moderately resistant strains of Pseudomonas aeruginosa. This antimicrobial effect becomes more pronounced with an increase in the concentration of the extract. Additionally, Pseudomonas aeruginosa that produce biofilms tend to consistently produce more Metallo-beta-lactamase (MBL), which can further contribute to antibiotic resistance.

Genotypic characterization of hypervirulent Klebsiella pneumoniae (hvKp) in a tertiary care Indian hospital.

HypervirulentKlebsiella pneumoniae(hvKp) is an emerging pathogen and causes endophthalmitis, liver abscess, osteomyelitis, meningitis, and necrotizing soft tissue infections in both immunodeficient and healthy people. The acquisition of the antibiotic resistance genes of hvKp has become an emerging concern throughout the globe. In this study, a total of 74K. pneumoniaeisolates were collected and identified by VITEK2 andblaSHVgene amplification. Out of these, 18.91% (14/74) isolates were identified as hvKp by both phenotypic string test and genotypiciucAPCR amplification. The antibiotic susceptibility revealed that 57.14% (8/14) isolates were multidrug-resistant (MDR) and 35.71% (5/14) isolates were extremely drug-resistant (XDR). All the isolates were resistant to β-lactam, β-lactamase + inhibitor groups of antibiotics, and the least resistance to colistin. Of 14 hvKp isolates, all isolates are positive for iroB(100%),followed byiutA(92.85%),peg344(85.71%), rmpA (57.14%), and magA(21.42%) genes. Among serotypes, K1 was the most prevalent serotype 21.4% (3/14), followed by K5 14.3% (2/14). The most common carbapenemase gene was blaOXA-48 (78.57%) followed by blaNDM (14.28%) and blaKPC (14.28%) which co-carried multiple resistance genes such as blaSHV (100%), blaCTX-M (92.85%), and blaTEM (78.57%). About 92.85% (13/14) of hvKp isolates were strong biofilm producers, while one isolate (hvKp 10) was the only moderate biofilm producer. The (GTG)5-PCR molecular typing method revealed high diversity among the hvKp isolates in the tertiary care hospital. Our findings suggest that MDR-hvKp is an emerging pathogen and a challenge for clinical practice. In order to avoid hvKp strain outbreaks in hospital settings, robust infection control and effective surveillance should be implemented.

Evaluation of Biofilm Production and Antifungal Susceptibility to Fluconazole in Clinical Isolates of Candida spp. in Both Planktonic and Biofilm Form.

Candida spp. are an important opportunistic pathogen that can represent a possible cause of severe infections, especially in immunocompromised individuals. The clinical impact of Candida spp. depends, in part, on the ability to form biofilms, communities of nestled cells into the extracellular matrix. In this study, we compared the biofilm formation ability of 83 strains of Candida spp. isolated from blood cultures and other materials, such as respiratory samples, urine, and exudate, and their sensitivity to fluconazole (FLZ). Strains were divided into tertiles to establish cut-offs to classify isolates as low, moderate, or high biofilm producers (<0.26, 0.266-0.839, >0.839) and biofilms with low, moderate, or high metabolic activity (<0.053, 0.053-0.183, >0.183). A non-linear relationship between biofilm production and metabolic activity was found in C. glabrata and C. tropicalis. In addition, the increase in minimum biofilm eradication concentrations (MBEC50) compared to the Minor Inhibitory Concentration (PMIC) of the planktonic form in Candida spp. confirms the role of biofilm in the induction of resistance to FLZ.

Genotypic investigation of drugs resistance and biofilm genes among ciprofloxacin-resistant Pseudomonas aeruginosa isolated from burn patients

Burn patients are the serious targets of hospital-acquired infection occurred by Pseudomonas aeruginosa ( P. aeruginosa) , which is a main cause of burn patients morbidity and mortality, so this study aimed to molecular investigation of genes associated with antibiotic resistance among ciprofloxacin-resistant Pseudomonas aeruginosa. Current study involved 168 burn patients involved 107(63.69%) and 61 (36.3%) were females and male respectively, the results showed 132(78.57%) gram negative bacteria and 27(16.0%) gram positive bacteria while 9(5.3%) no bacterial growth. The results of Vitek-2 compact system recorded 75(44.64%) as P. aeruginosa isolates. Results of ciprofloxacin susceptibility showed 29(38.67%), while 34(45.33%) and 12(16%) of isolates were intermediate and sensitive respectively. Congo red agar assay was applied in current study for qualitative evaluation of pathogenic biofilm were 23/29 (79.31%), and 6/26(20.86%) as high, and moderate biofilm producer among isolates respectively. Result of Polymerase chain reaction (PCR) showed that gyrB was recorded among ciprofloxacin-resistant P. aeruginosa at rate 28/29 (96.55%). At same respect. results of efflux pump genes for oqxA and oqxB genes were detected among this pathogen at rate 26/29 (89.65%) and 29/29(100%) respectively. outer membrane genes, revealed that mexR gene was 27/29 (93.1%), while the spreading of the oprD gene was 26/29(89.65%). Results of molecular detection about colistin resistance genes revealed that the mcr-3 gene was 8/29 (27.58%), but genes of mcr-1 and mcr-2 were no detected in this work, Finally, results showed that 23/29 (79.31%) of ciprofloxacin-resistant P. aeruginosa were harbored pml gene.

Apple cider vinegar exhibits promising antibiofilm activity against multidrug-resistant Bacillus cereus isolated from meat and their products.

Bacillus cereus (B. cereus) biofilm is grown not only on medical devices but also on different substrata and is considered a potential hazard in the food industry. Quorum sensing plays a serious role in the synthesis of biofilm with its surrounding extracellular matrix enabling irreversible connection of the bacteria. The goal of the current investigation was to ascertain the prevalence, patterns of antimicrobial resistance, and capacity for B. cereus biofilm formation in meat and meat products in Egypt. In all, 150 meat and meat product samples were used in this study. For additional bacteriological analysis, the samples were moved to the Bacteriology Laboratory. Thereafter, the antimicrobial, antiquorum sensing, and antibiofilm potential of apple cider vinegar (ACV) on B. cereus were evaluated. Out of 150 samples, 34 (22.67%) tested positive for B. cereus. According to tests for antimicrobial susceptibility, every B. cereus isolates tested positive for colistin and ampicillin but negative for ciprofloxacin and imipenem. The ability to form biofilms was present in all 12 multidrug-resistant B. cereus isolates (n = 12); of these, 6 (50%), 3 (25%), and 3 (25%) isolates were weak, moderate, and strong biofilm producers, respectively. It is noteworthy that the ACV demonstrated significant inhibitory effects on B. cereus isolates, with minimum inhibitory concentrations varying between 2 and 8 μg/ml. Furthermore, after exposing biofilm-producing B. cereus isolates to the minimum biofilm inhibitory concentrations 50 of 4 μg/ml, it demonstrated good antibiofilm activity (>50% reduction of biofilm formation). Strong biofilm producers had down-regulated biofilm genes (tasA and sipW) and their regulator (plcR) compared to the control group, according to reverse transcriptase quantitative polymerase chain reaction analysis. Our study is the first report, that spotlights the ACV activity against B. cereus biofilm and its consequence as a strong antibacterial and antibiofilm agent in the food industry and human health risk.