scholarly journals Investigation on Antibiotic-Resistance, Biofilm Formation and Virulence Factors in Multi Drug Resistant and Non Multi Drug Resistant Staphylococcus pseudintermedius

2019 ◽  
Vol 7 (12) ◽  
pp. 702 ◽  
Author(s):  
Gabriele Meroni ◽  
Joel F. Soares Filipe ◽  
Lorenzo Drago ◽  
Piera A. Martino
Keyword(s):  
Antibiotic Resistance ◽  
Virulence Factors ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Drug Resistant ◽  
Virulence Determinants ◽  
Commensal Bacterium ◽  
Staphylococcus Pseudintermedius ◽  
Microtiter Plate Assay ◽  
Human Agent

Staphylococcus pseudintermedius is a commensal bacterium frequently isolated from canine skin and recognized as a zoonotic agent especially for dog-owners. This study focused on (a) the antibiotic-resistance phenotypes; (b) the ability to produce biofilm (slime); and (c) the dissemination of virulence factors in S. pseudintermedius strains. Seventy-three S. pseudintermedius strains were screened for antibiotic-resistance against 22 different molecules by means of Kirby-Bauer assay. The ability to produce biofilm was investigated using the microtiter plate assay (MtP) and the amplification of icaA and icaD genes. Virulence factors such as cytotoxins (lukI), enterotoxins (seC), and exfoliative toxins (siet, expA, and expB) were evaluated. The antibiotic-resistance profiles revealed 42/73 (57%) multi-drug resistant (MDR) strains and 31/73 (43%) not-MDR. All the MDR strains and 8/31 (27%) of not-MDR resulted in biofilm producers. Leukotoxin LukI was found in 70/73 (96%) of the isolates. Moreover, the enterotoxin gene seC was detected in 47/73 (64%) of the strains. All the isolates carried the siet gene, whereas expA and expB were found in 3/73 (4%) and 5/73 (7%), respectively. In conclusion, S. pseudintermedius should be considered a potential zoonotic and human agent able to carry different virulence determinants and capable of producing biofilm which facilitates horizontal gene transfer.

Evaluation of the Role of Staphylococci in the Pathomechanism of Conjunctivitis

2021 ◽  
Author(s):  
Ewa Jasińska ◽  
Agnieszka Bogut ◽  
Agnieszka Magryś ◽  
Alina Olender
Keyword(s):  
Antibiotic Resistance ◽  
Epithelial Cells ◽  
Biofilm Formation ◽  
Methicillin Resistance ◽  
Microtiter Plate ◽  
Host Cells ◽  
Diffusion Method ◽  
Microtiter Plate Assay ◽  
Low Prevalence

Abstract Purpose: Determination of the association between ica genes and phenotypic biofilm formation in staphylococcal isolates involved in conjunctivitis, their antibiotic resistance as well as detection of selected virulence characteristics: adhesion to epithelial cells and in vitro cytotoxicity.Methods: The study included 26 Staphylococcus aureus (SA) and 26 Staphylococcus epidermidis (SE) isolates. The presence of icaAD genes and ica operon was determined by the PCR assay. Phenotypic biofilm formation was verified using the microtiter plate assay. Antibiotic resistance was performed using the disc diffusion method. Staphylococcal ability to attach to host cells was assessed by flow cytometry. Cytotoxicity on epithelial cells was evaluated by LDH assay.Results: The ica genes were detected in 26.9% of SE and in 42.3% of SA isolates. Only 15.3% of isolates (SE) were positive for both the icaAD and the ica operon. Phenotypically, 19.2% of SE isolates were strong biofilm producers, among which three were both icaAD- and ica operon-positive. 26.9% of SA isolates were strong biofilm producers. Methicillin resistance (MR) was detected in 34.6% of SE and 26.9% of SA isolates. 75% of MR isolates were multidrug resistant. SA isolates adhered to host cells more extensively than SE. SA isolates released higher level of LDH than SE.Conclusions: Adherence abilities were commonly observed in staphylococci associated with conjunctivitis. However, low prevalence of isolates positive for a complete and functional ica locus and low prevalence of strong biofilm producers was detected. SA adhered to a greater extent to eukaryotic cells than SE and were more cytotoxic.

scholarly journals Comparison of biofilm formation and efflux pumps in ESBL and carbapenemase producing Klebsiella pneumoniae

2018 ◽  
Vol 12 (03) ◽  
pp. 156-163 ◽  
Author(s):  
Burak Yazgan ◽  
Ibrahim Türkel ◽  
Rıdvan Güçkan ◽  
Kılınç Kılınç ◽  
Tuba Yıldırım
Keyword(s):  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Cell Communication ◽  
Opportunistic Pathogen ◽  
Efflux Pumps ◽  
Microtiter Plate ◽  
Virulence Determinants ◽  
Clinical Environment ◽  
Microtiter Plate Assay ◽  
Significant Difference

Introduction: Klebsiella pneumoniae is an opportunistic pathogen that causes a range of diseases. The appearance of extended-spectrum β-lactamase -and carbapenemase-producing strains, in addition to the biofilm-forming phenotype, is a major problem in the clinical environment. Methodology: A total of 33 clinical K. pneumoniae isolates were used in this study. Antimicrobial susceptibilities were assessed by a disc diffusion assay. Biofilm formation was determined by a microtiter plate assay, staining with 1% crystal violet and measuring  absorbance after destaining. Moreover, expression of acrA, kdeA, ketM, kpnEF, and kexD efflux associated genes  was measured by qRT-PCR. Results: Isolates displayed high resistance to β-lactams such as cefazolin, cefuroxime, ceftriaxone, cefepime, piperacillin-tazobactam, imipenem, and meropenem and decreased resistance to gentamicin, amikacin, ciprofloxacin, and levofloxacin. ESBL-producing isolates formed more biofilm than carbapenemase-producing isolates. The mRNA expression levels in KPC isolates for acrA (2-fold), kdeA (2.7-fold), ketM (2.2-fold), and kpnEF (3.4-fold) were significantly increased compared to ESBL-producing isolates. There was no significant difference in kexD expression level. Conclusions: Under the conditions used here ESBL-producing isolates formed more biofilm than KPC postive isolates; this was associated with virulence determinants which were also transferred by plasmids together with ESBLs enzymes. Moreover, the upregulation of acrA, kdeA, ketM, and kpnEF efflux pumps was seen in carbapenemase-producing isolates demonstrating that high expression of efflux pumps alone could not confer resistance but may act as a physiological determinant such as bacterial pathogenicity and virulence, and cell-to-cell communication for bacteria.

scholarly journals No Correlation between Biofilm Formation, Virulence Factors, and Antibiotic Resistance in Pseudomonas aeruginosa: Results from a Laboratory-Based In Vitro Study

Antibiotics ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1134
Author(s):  
Márió Gajdács ◽  
Zoltán Baráth ◽  
Krisztina Kárpáti ◽  
Dóra Szabó ◽  
Donatella Usai ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibiotic Resistance ◽  
Virulence Factors ◽  
Biofilm Formation ◽  
Bacterial Infections ◽  
Pearson Correlation ◽  
Pigment Production ◽  
Virulence Determinants ◽  
Secretion Systems

Pseudomonas aeruginosa (P. aeruginosa) possesses a plethora of virulence determinants, including the production of biofilm, pigments, exotoxins, proteases, flagella, and secretion systems. The aim of our present study was to establish the relationship between biofilm-forming capacity, the expression of some important virulence factors, and the multidrug-resistant (MDR) phenotype in P. aeruginosa. A total of three hundred and two (n = 302) isolates were included in this study. Antimicrobial susceptibility testing and phenotypic detection of resistance determinants were carried out; based on these results, isolates were grouped into distinct resistotypes and multiple antibiotic resistance (MAR) indices were calculated. The capacity of isolates to produce biofilm was assessed using a crystal violet microtiter-plate based method. Motility (swimming, swarming, and twitching) and pigment-production (pyoverdine and pyocyanin) were also measured. Pearson correlation coefficients (r) were calculated to determine for antimicrobial resistance, biofilm-formation, and expression of other virulence factors. Resistance rates were the highest for ceftazidime (56.95%; n = 172), levofloxacin (54.97%; n = 166), and ciprofloxacin (54.64%; n = 159), while lowest for colistin (1.66%; n = 5); 44.04% (n = 133) of isolates were classified as MDR. 19.87% (n = 60), 20.86% (n = 63) and 59.27% (n = 179) were classified as weak, moderate, and strong biofilm producers, respectively. With the exception of pyocyanin production (0.371 ± 0.193 vs. non-MDR: 0.319 ± 0.191; p = 0.018), MDR and non-MDR isolates did not show significant differences in expression of virulence factors. Additionally, no relevant correlations were seen between the rate of biofilm formation, pigment production, or motility. Data on interplay between the presence and mechanisms of drug resistance with those of biofilm formation and virulence is crucial to address chronic bacterial infections and to provide strategies for their management.

scholarly journals Detection of biofilm formation and correlation with antibiotic resistance in Acinetobacter baumannii

2020 ◽  
Vol 11 (SPL4) ◽  
pp. 2474-2480
Author(s):  
Anitha M ◽  
Jagatheeswary P A T ◽  
Sumathi G ◽  
Kamini B
Keyword(s):  
Antibiotic Resistance ◽  
Acinetobacter Baumannii ◽  
Biofilm Formation ◽  
Screening Method ◽  
Strong Relationship ◽  
Microtiter Plate ◽  
Diffusion Method ◽  
Drug Resistant ◽  
Biofilm Production ◽  
Hospital Acquired

Acinetobacter baumannii is a major cause of hospital acquired infections worldwide and is associated with resistance to routinely used antibiotics. Many clinical isolates of Acinetobacter baumannii  are found to be biofilm producers. Hence there is difficulty in treating patients with Multi Drug Resistant Acinetobacter baumannii  (MDRAB). This present study aimed to study antibiotic resistance of A. baumannii isolates  and to evaluate the biofilm formation of Acinetobacter baumannii  by Tube Method (TM) and Microtiter Plate Method(MTPM).In this study, 73 A. baumannii  isolates of various clinical specimens were evaluated. Confirmation was done through conventional methods. Testing for antimicrobial susceptibility was done by Kirby Bauer disc diffusion method. Biofilm formation was studied by TM and MTPM. Of the 73 isolates, 26(36%) were from urine, 19 (26%) from pus, 17 (23%) from sputum and 11 (15%) from other miscellaneous(body fluids excluding blood), out of which 81%(59/73) isolates were Multi Drug Resistant (MDR). 63% and 84% of isolates showed biofilm production in TM and MTPM, respectively. When comparing these two methods, MTPM assay was better than TM. Presence of a strong relationship between biofilm formation and MDRAB has been confirmed by the present study. Both methods used for detection of biofilm formation were found to be statistically significant. Sensitivity of MTPM was more than TM, which is supported by higher Positive Predictive Value of 87.5%. Therefore MTPM is a better method than TM and can be used as a screening method to detect biofilm production.

scholarly journals Virulence Factors in Coagulase-Negative Staphylococci

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 170
Author(s):  
Angela França ◽  
Vânia Gaio ◽  
Nathalie Lopes ◽  
Luís D. R. Melo
Keyword(s):  
Antibiotic Resistance ◽  
Virulence Factors ◽  
Resistance Genes ◽  
Biofilm Formation ◽  
Antibiotic Resistance Genes ◽  
Coagulase Negative Staphylococci ◽  
Healthcare Associated ◽  
Major Pathogens

Coagulase-negative staphylococci (CoNS) have emerged as major pathogens in healthcare-associated facilities, being S. epidermidis, S. haemolyticus and, more recently, S. lugdunensis, the most clinically relevant species. Despite being less virulent than the well-studied pathogen S. aureus, the number of CoNS strains sequenced is constantly increasing and, with that, the number of virulence factors identified in those strains. In this regard, biofilm formation is considered the most important. Besides virulence factors, the presence of several antibiotic-resistance genes identified in CoNS is worrisome and makes treatment very challenging. In this review, we analyzed the different aspects involved in CoNS virulence and their impact on health and food.

scholarly journals Draft Genome Sequence of a Clinically Isolated Extensively Drug-Resistant Pseudomonas aeruginosa Strain

2016 ◽  
Vol 4 (2) ◽  
Author(s):  
Bhavani Manivannan ◽  
Niranjana Mahalingam ◽  
Sudhir Jadhao ◽  
Amrita Mishra ◽  
Pravin Nilawe ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibiotic Resistance ◽  
Genome Assembly ◽  
Biofilm Formation ◽  
Draft Genome ◽  
Drug Resistant ◽  
Draft Genome Assembly ◽  
Extensively Drug Resistant ◽  
History Of ◽  
Urinary Tuberculosis

We present the draft genome assembly of an extensively drug-resistant (XDR) Pseudomonas aeruginosa strain isolated from a patient with a history of genito urinary tuberculosis. The draft genome is 7,022,546 bp with a G+C content of 65.48%. It carries 7 phage genomes, genes for quorum sensing, biofilm formation, virulence, and antibiotic resistance.

scholarly journals Effect of S-PRG Eluate on Biofilm Formation and Enzyme Activity of Oral Bacteria

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Masahiro Yoneda ◽  
Nao Suzuki ◽  
Yosuke Masuo ◽  
Akie Fujimoto ◽  
Kosaku Iha ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Composite Resin ◽  
Fusobacterium Nucleatum ◽  
Microtiter Plate ◽  
Oral Bacteria ◽  
Gelatin Film ◽  
Glass Ionomer ◽  
Microtiter Plate Assay ◽  
Substrate Hydrolysis ◽  
Adherence Ability

Recently, the antibacterial activity of a composite resin containing prereacted glass ionomer (S-PRG) filler was revealed. We examined the effect of an S-PRG eluate on various biologic activities ofStreptococcus mutansandPorphyromonas gingivalis. Adherence ability ofS. mutanswas evaluated by microtiter plate assay; protease and gelatinase activities ofP. gingivaliswere examined by synthetic substrate hydrolysis and gelatin film spot assay, respectively. Coaggregation ofP. gingivaliswithFusobacterium nucleatumwas also examined. S-PRG eluate was found to suppress streptococcal adherence. S-PRG eluate inhibited the protease and gelatinase activities ofP. gingivalisand the coaggregation betweenP. gingivalisandF. nucleatum. These results indicate that S-PRG eluate suppresses streptococcal adherence and inhibits the protease and coaggregation activities ofP. gingivalis. These findings may prompt research into novel strategies for preventing caries and periodontitis.

scholarly journals High-Quality Draft Genome Sequence of the Multidrug-Resistant Clinical Isolate Enterococcus faecium VRE16

2016 ◽  
Vol 4 (5) ◽  
Author(s):  
Suelen Scarpa de Mello ◽  
Daria Van Tyne ◽  
Andrei Nicoli Gebieluca Dabul ◽  
Michael S. Gilmore ◽  
Ilana L. B. C. Camargo
Keyword(s):  
Antibiotic Resistance ◽  
Virulence Factors ◽  
Genome Sequence ◽  
Enterococcus Faecium ◽  
Draft Genome ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Draft Genome Sequence ◽  
Commensal Bacterium ◽  
High Quality

Specific lineages of the commensal bacterium Enterococcus faecium belonging to CC17, especially ST412, have been isolated from patients in several hospitals worldwide and harbor antibiotic resistance genes and virulence factors. Here, we report a high-quality draft genome sequence and highlight features of E. faecium VRE16, a representative of this ST.

scholarly journals Relationship between type III secretion toxins, biofilm formation, and antibiotic resistance in clinical Pseudomonas aeruginosa isolates

2021 ◽  
Vol 11 (6) ◽  
pp. 1075-1082
Author(s):  
S. Derakhshan ◽  
A. Rezaee ◽  
Sh. Mohammadi
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibiotic Resistance ◽  
Multidrug Resistance ◽  
Type Iii Secretion ◽  
Biofilm Formation ◽  
Virulence Gene ◽  
Microtiter Plate ◽  
P Value ◽  
Type Iii ◽  
Disk Diffusion Assay

Background and aim. Pseudomonas aeruginosa is considered as a notorious pathogen due to its multidrug resistance and life threatening infections. We investigated the relationship between type III secretion toxins, biofilm formation, and antibiotic resistance among clinical P. aeruginosa isolates. Methods. A total of 70 genetically distinct clinical P. aeruginosa isolates were characterized for antibiotic resistance by disk diffusion assay. Biofilm formation was evaluated by microtiter plate method and presence of four exo genes (exoS, exoU, exoT and exoY) was investigated by PCR. A p-value < 0.05 was regarded statistically significant. Results. The most effective antibiotics were Meropenem and Piperacillin. Multidrug resistance was more prevalent in the ciprofloxacin-resistant isolates than in the susceptible isolates. The most frequently identified exo was exoS (37.1%). Genotype exoS/exoT was found in 4 isolates, while genotype exoU/exoT was not found. Prevalence of exoS was generally higher in the susceptible isolates than in the resistant isolates. A significant association was found between the formation of strong biofilm and resistance to antibiotics (p < 0.05). Prevalence of exoY and exoU was higher in the non-strong biofilm producers compared to the strong biofilm producers. Conclusion. Our study revealed formation of strong biofilm along with antibiotic resistance and the presence of exo genes in P. aeruginosa isolates. Knowledge of virulence gene profiles and biofilm formation may be useful in deciding appropriate treatment.

scholarly journals Virulence factors and antibiotic resistance in enterococci isolated from food-stuffs

2011 ◽  
Vol 56 (No. 7) ◽  
pp. 352-357 ◽  
Author(s):  
K. Trivedi ◽  
S. Cupakova ◽  
R. Karpiskova
Keyword(s):  
Antibiotic Resistance ◽  
Virulence Factors ◽  
Diffusion Method ◽  
Haemolytic Activity ◽  
Virulence Determinants ◽  
Disk Diffusion Method ◽  
Specific Pcr ◽  
Virulence Potential ◽  
Enterococcal Species ◽  
Species Specific

A collection of 250 enterococci isolated from various food-stuffs were used to investigate seven virulence determinants and the microbial susceptibility of eight antibiotics. Species-specific PCR revealed the presence of E. faecalis (127 isolates), E. faecium (77 isolates), E. casseliflavus (21 isolates), E. mundtii (19 isolates) and E. durans (six isolates). Multiplex PCR for virulence factors showed that from a total 250 isolates, 221 (88.4%) carried one or more virulence-encoding genes. &beta;-Haemolytic activity was also evident in enterococcal species other than E. faecalis and E. faecium. Species other than E. faecalis and E. faecium isolated from food are also seen to harbour the potential for virulence. Antimicrobial susceptibility testing using the disk diffusion method showed that of the total 250 isolates, 114 (46%) were resistant to cephalothin and 94 (38%) to ofloxacin. Lower antibiotic resistance was seen with ampicillin, chloramphenicol, gentamicin and teicoplanin. None of the isolates was found to be resistant to vancomycin. The results of this study show that food can play an important role in the spread of enterococci with virulence potential through the food chain to the human population.

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