Methicillin resistance genes and in vitro biofilm formation among Staphylococcus aureus isolates from bovine mastitis in India

2019 ◽  
Vol 64 ◽  
pp. 117-124 ◽  
Author(s):  
Muneeba Shafi Shah ◽  
Sabia Qureshi ◽  
Zahid Kashoo ◽  
Shaheen Farooq ◽  
Shakil Ahmad Wani ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Resistance Genes ◽  
Biofilm Formation ◽  
Methicillin Resistance ◽  
Bovine Mastitis

scholarly journals Role of Biofilm-Associated Protein Bap in the Pathogenesis of Bovine Staphylococcus aureus

2004 ◽  
Vol 72 (4) ◽  
pp. 2177-2185 ◽  
Author(s):  
Carme Cucarella ◽  
M. Ángeles Tormo ◽  
Carles Úbeda ◽  
M. Pilar Trotonda ◽  
Marta Monzón ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Bovine Mastitis ◽  
Serum Samples ◽  
Group 3 ◽  
Group 2 ◽  
Vitro Analysis ◽  
Group 1

ABSTRACT Staphylococcus aureus is a common cause of intramammary infections, which frequently become chronic, associated with the ability of the bacteria to produce biofilm. Here, we report a relationship between the ability to produce chronic bovine mastitis and biofilm formation. We have classified bovine mastitis S. aureus isolates into three groups based on the presence of particular genetic elements required for biofilm formation: group 1 (ica + bap +), group 2 (ica +, bap negative), and group 3 (ica negative, bap negative). Overall, animals naturally infected with group 1 and 2 isolates had a lower milk somatic cell count than those infected with isolates of group 3. In addition, Bap-positive isolates were significantly more able to colonize and persist in the bovine mammary gland in vivo and were less susceptible to antibiotic treatments when forming biofilms in vitro. Analysis of the structural bap gene revealed the existence of alternate forms of expression of the Bap protein in S. aureus isolates obtained under field conditions throughout the animal's life. The presence of anti-Bap antibodies in serum samples taken from animals with confirmed S. aureus infections indicated the production of Bap during infection. Furthermore, disruption of the ica operon in a bap-positive strain had no effect on in vitro biofilm formation, a finding which strongly suggested that Bap could compensate for the deficiency of the PIA/PNAG product (a biofilm matrix polysaccharide). Altogether, these results demonstrate that, in the bovine intramammary gland, the presence of Bap may facilitate a biofilm formation connected with the persistence of S. aureus.

scholarly journals Paracetamol modulates biofilm formation in Staphylococcus aureus clonal complex 8 strains

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Andi R. Sultan ◽  
Kirby R. Lattwein ◽  
Nicole A. Lemmens-den Toom ◽  
Susan V. Snijders ◽  
Klazina Kooiman ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Fluorescent Protein ◽  
Clonal Complex ◽  
Regulatory Pathways ◽  
Immune Modulator ◽  
Viability Assay ◽  
Analgesic Agents ◽  
Green Fluorescent

AbstractStaphylococcus aureus biofilms are a major problem in modern healthcare due to their resistance to immune system defenses and antibiotic treatments. Certain analgesic agents are able to modulate S. aureus biofilm formation, but currently no evidence exists if paracetamol, often combined with antibiotic treatment, also has this effect. Therefore, we aimed to investigate if paracetamol can modulate S. aureus biofilm formation. Considering that certain regulatory pathways for biofilm formation and virulence factor production by S. aureus are linked, we further investigated the effect of paracetamol on immune modulator production. The in vitro biofilm mass of 21 S. aureus strains from 9 genetic backgrounds was measured in the presence of paracetamol. Based on biofilm mass quantity, we further investigated paracetamol-induced biofilm alterations using a bacterial viability assay combined with N-Acetylglucosamine staining. Isothermal microcalorimetry was used to monitor the effect of paracetamol on bacterial metabolism within biofilms and green fluorescent protein (GFP) promoter fusion technology for transcription of staphylococcal complement inhibitor (SCIN). Clinically relevant concentrations of paracetamol enhanced biofilm formation particularly among strains belonging to clonal complex 8 (CC8), but had minimal effect on S. aureus planktonic growth. The increase of biofilm mass can be attributed to the marked increase of N-Acetylglucosamine containing components of the extracellular matrix, presumably polysaccharide intercellular adhesion. Biofilms of RN6390A (CC8) showed a significant increase in the immune modulator SCIN transcription during co-incubation with low concentrations of paracetamol. Our data indicate that paracetamol can enhance biofilm formation. The clinical relevance needs to be further investigated.

scholarly journals The First Report of a Methicillin-Resistant Staphylococcus aureus Isolate Harboring Type IV SCCmec in Thailand

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 430
Author(s):  
Wichai Santimaleeworagun ◽  
Praewdow Preechachuawong ◽  
Wandee Samret ◽  
Tossawan Jitwasinkul
Keyword(s):  
Staphylococcus Aureus ◽  
Resistance Genes ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Sccmec Type ◽  
Clinical Strain ◽  
Spa Typing ◽  
Methicillin Resistant ◽  
Type Iv ◽  
Antimicrobial Resistance Genes

Methicillin-resistant Staphylococcus aureus (MRSA) is mostly found in Thailand in the hospital as a nosocomial pathogen. This study aimed to report the genetic characterization of a clinical community-acquired MRSA (CA-MRSA) isolate collected from hospitalized patients in Thailand. Among 26 MRSA isolates, S. aureus no. S17 preliminarily displayed the presence of a staphylococcal cassette chromosome mec (SCCmec) type IV pattern. The bacterial genomic DNA was subjected to whole-genome sequencing. Panton–Valentine leukocidin (PVL) production, virulence toxins, and antibiotic resistance genes were identified, and multi-locus sequence typing (MLST) and spa typing were performed. The strain was matched by sequence to MLST type 2885 and spa type t13880. This strain carried type IV SCCmec with no PVL production. Five acquired antimicrobial resistance genes, namely blaZ, mecA, Inu(A), tet(K), and dfrG conferring resistance to β-lactams, lincosamides, tetracycline, and trimethoprim, were identified. The detected toxins were exfoliative toxin A, gamma-hemolysin, leukocidin D, and leukocidin E. Moreover, there were differences in seven regions in CR-MRSA no. S17 compared to CA-MRSA type 300. In summary, we have reported the ST2885-SCCmec IV CA-MRSA clinical strain in Thailand for the first time, highlighting the problem of methicillin resistance in community settings and the consideration in choosing appropriate antibiotic therapy.

scholarly journals Anaerobic Conditions Induce Expression of Polysaccharide Intercellular Adhesin in Staphylococcus aureus and Staphylococcus epidermidis

2001 ◽  
Vol 69 (6) ◽  
pp. 4079-4085 ◽  
Author(s):  
Sarah E. Cramton ◽  
Martina Ulrich ◽  
Friedrich Götz ◽  
Gerd Döring
Keyword(s):  
Staphylococcus Aureus ◽  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Extracellular Polysaccharide ◽  
Growth Conditions ◽  
Anaerobic Conditions ◽  
Anaerobic Environment ◽  
Specific Mrna

ABSTRACT Products of the intercellular adhesion (ica) operon in Staphylococcus aureus and Staphylococcus epidermidis synthesize a linear β-1,6-linked glucosaminylglycan. This extracellular polysaccharide mediates bacterial cell-cell adhesion and is required for biofilm formation, which is thought to increase the virulence of both pathogens in association with prosthetic biomedical implants. The environmental signal(s) that triggers ica gene product and polysaccharide expression is unknown. Here we demonstrate that anaerobic in vitro growth conditions lead to increased polysaccharide expression in both S. aureus and S. epidermidis, although the regulation is less stringent inS. epidermidis. Anaerobiosis also dramatically stimulates ica-specific mRNA expression inica- and polysaccharide-positive strains of both S. aureus and S. epidermidis.These data suggest a mechanism whereby ica gene expression and polysaccharide production may act as a virulence factor in an anaerobic environment in vivo.

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 Isolation of Biofilm Producing Methicillin-Resistant Staphylococcus aureus from Hospitalized Orthopaedic Patients in Kano State, Nigeria

2021 ◽  
Vol 28 (1) ◽  
pp. 66-74
Author(s):  
D.A. Oche ◽  
U. Abdulrahim ◽  
A.S. Oheagbulem ◽  
B.O. Olayinka
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Community Settings ◽  
Potential Threat ◽  
Orthopaedic Patients ◽  
Nasal Swabs ◽  
Public Health Challenge

Biofilm formation and resistance to methicillin are among the factors that makes Staphylococcus aureus a very important human pathogen in both health-care and community settings. This study investigated methicillin-resistance among biofilm-producing S. aureus isolated from 49 orthopaedic in-patients within a 3 months period. Wound swabs, nasal swabs, bed swabs and urine samples were collected from each patient. The samples were cultured and screened for presence of S. aureus while the micro-titre plate method was used to detect biofilm producing isolates. PCR technique was finally used to detect the presence of mecA gene in methicilin resistant S. aureus (MRSA) isolates. Findings reveal 14.8% of bacterial isolates were Staphylococcus aureus of which 96.4% were biofilm-producers. However, strong biofilm producers constitute 11.1%. The mecA gene was detected in 15.8% of the MRSA isolates. Therefore, MRSA among biofilm-producing S. aureus is a potential threat primarily to the community of National Orthopaedic Hospital Dala and a major public health challenge. Keywords: Biofilm, Methicillin-resistance Staphylococcus aureus (MRSA), mecA gene, Orthopaedic patient

scholarly journals Expression of icaA and icaD genes in biofilm formation in Staphylococcus aureus isolates from bovine subclinical mastitis

2021 ◽  
Vol 41 ◽  
Author(s):  
Viviane F. Marques ◽  
Huarrisson A. Santos ◽  
Thomas H. Santos ◽  
Dayanne A. Melo ◽  
Shana M.O. Coelho ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Phenotypic Expression ◽  
Bovine Mastitis ◽  
Subclinical Mastitis ◽  
Transcriptional Analysis ◽  
Important Species ◽  
High Prevalence ◽  
Staphylococcus Spp

ABSTRACT: Staphylococcus spp. plays a significant role in the etiology of bovine mastitis. Staphylococcus aureus is considered the most important species due to the high prevalence and the difficulty of in vivo treatment that is related to the expression of virulence factors and biofilm formation. This study aimed to detect the phenotypic expression of the biofilm formation in 20 S. aureus isolated from bovine mastitis and to evaluate the expression and regulation of genes involved in its production. MALDI-TOF and phenogenotypic identification assays were performed to characterize the isolates. The phenotypic biofilm production and the presence of icaA and icaD and bap genes were evaluated. The Agr system was typified (agr I, agr II, agr III and agr IV) and its regulator (agr RNAIII) was detected. Furtherly, Real-time PCR (qPCR) was performed at chosen times to quantify the expression of icaA, icaD and hld genes in three selected isolates. All 20 strains were biofilm producers and most presented icaA and icaD genes. Only one isolate presented the bap gene. The agr gene type II showed a prevalence of 70%. Transcriptional analysis revealed increased expression of ica genes at eight hours of growth. These results confirm that polysaccharides production mediated by the icaADBC operon genes is an essential mechanism to the biofilm formation and contributes to the early stages of bacterial growth.

scholarly journals Within-Host Adaptation of Staphylococcus aureus in a Bovine Mastitis Infection Is Associated with Increased Cytotoxicity

2021 ◽  
Vol 22 (16) ◽  
pp. 8840
Author(s):  
Katharina Mayer ◽  
Martin Kucklick ◽  
Helene Marbach ◽  
Monika Ehling-Schulz ◽  
Susanne Engelmann ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Bovine Mastitis ◽  
Adaptive Strategy ◽  
Subclinical Mastitis ◽  
Growth Conditions ◽  
Typical Feature ◽  
Host Adaptation ◽  
Bovine Mammary Epithelial Cells ◽  
Strain Pair

Within-host adaptation is a typical feature of chronic, persistent Staphylococcus aureus infections. Research projects addressing adaptive changes due to bacterial in-host evolution increase our understanding of the pathogen’s strategies to survive and persist for a long time in various hosts such as human and bovine. In this study, we investigated the adaptive processes of S. aureus during chronic, persistent bovine mastitis using a previously isolated isogenic strain pair from a dairy cow with chronic, subclinical mastitis, in which the last variant (host-adapted, Sigma factor SigB-deficient) quickly replaced the initial, dominant variant. The strain pair was cultivated under specific in vitro infection-relevant growth-limiting conditions (iron-depleted RPMI under oxygen limitation). We used a combinatory approach of surfaceomics, molecular spectroscopic fingerprinting and in vitro phenotypic assays. Cellular cytotoxicity assays using red blood cells and bovine mammary epithelial cells (MAC-T) revealed changes towards a more cytotoxic phenotype in the host-adapted isolate with an increased alpha-hemolysin (α-toxin) secretion, suggesting an improved capacity to penetrate and disseminate the udder tissue. Our results foster the hypothesis that within-host evolved SigB-deficiency favours extracellular persistence in S. aureus infections. Here, we provide new insights into one possible adaptive strategy employed by S. aureus during chronic, bovine mastitis, and we emphasise the need to analyse genotype–phenotype associations under different infection-relevant growth conditions.

scholarly journals Bacteriophage-Derived PeptidaseCHAPKEliminates and Prevents Staphylococcal Biofilms

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Mark Fenton ◽  
Ruth Keary ◽  
Olivia McAuliffe ◽  
R. Paul Ross ◽  
Jim O'Mahony ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Antibacterial Agents ◽  
Antimicrobial Agents ◽  
Bovine Mastitis ◽  
Economic Losses ◽  
Food Sector ◽  
Staphylococcal Infections ◽  
Prevention And Treatment

New antibacterial agents are urgently needed for the elimination of biofilm-forming bacteria that are highly resistant to traditional antimicrobial agents. Proliferation of such bacteria can lead to significant economic losses in the agri-food sector. This study demonstrates the potential of the bacteriophage-derived peptidase,CHAPK, as a biocidal agent for the rapid disruption of biofilm-forming staphylococci, commonly associated with bovine mastitis. PurifiedCHAPKapplied to biofilms ofStaphylococcus aureusDPC5246 completely eliminated the staphylococcal biofilms within 4 h. In addition,CHAPKwas able to prevent biofilm formation by this strain. TheCHAPKlysin also reducedS. aureusin a skin decolonization model. Our data demonstrates the potential ofCHAPKas a biocidal agent for prevention and treatment of biofilm-associated staphylococcal infections or as a decontaminating agent in the food and healthcare sectors.

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