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.

scholarly journals Bovine mastitis: prevalence and antimicrobial susceptibility profile and detection of genes associated with biofilm formation in Staphylococcus aureus

2016 ◽  
Vol 37 (3) ◽  
pp. 1369
Author(s):  
Valeska Paula Casanova ◽  
Juceam Appio ◽  
Eduardo Kohl ◽  
Tais Regina Michaelsen ◽  
Daniel Santos Paim ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Susceptibility ◽  
Biofilm Formation ◽  
Antimicrobial Agents ◽  
Bovine Mastitis ◽  
Antimicrobial Susceptibility Profile ◽  
Nocardia Sp ◽  
Polymerase Chain ◽  
Safe Food ◽  
Corynebacterium Sp

Brazil currently ranks as one of the world leaders in food production and exportation. This scenario encourages the development of animal and plant health programs to ensure the production of safe food, helping the country to become an international provider of food for excellence. However, some health problems in dairy production, such as mastitis, have garnered increasing concern. This study aimed to estimate the prevalence of bovine mastitis in select properties located in the western Santa Catarina region, to assess the susceptibility profile to antimicrobial agents used for treatment and to check for the presence of genes (icaA and icaD) associated with biofilm formation in Staphylococcus aureus. In 148 milk samples collected, 72.97% had bacterial growth (n = 108). Among the isolated microorganisms, 21.62% (n = 32) were classified as Staphylococcus aureus, 18.91% (n = 28) as Staphylococcus sp. coagulase negative, 7.43% (n = 11) as Corynebacterium sp., 6.76% (n = 10) as Staphylococcus sp. coagulase positive, 5.41% (n = 8) as Nocardia sp. and 12.83% (n = 19) classified in different bacterial genera. Among the isolates submitted for antimicrobial susceptibility testing, it was observed that 8.95% (n = 6/67) had resistance to amoxicillin, 8.04% (n = 7/87) to ampicillin, 5.88% (n = 5/85) to cephalothin, 3.40% (n = 3/88) to ceftiofur and enrofloxacin, 20.45% (n = 18/88) to streptomycin, 17.04% (n = 15/88) to gentamicin and lincomycin, 31.81% (n = 28/88) to neomycin, 14.94% (n = 13/87) to penicillin and 25% (n = 22/88) to tetracycline. Staphylococcus sp. coagulase negative isolates showed higher multidrug resistance when compared to those of S. aureus and Staphylococcus sp. coagulase positive. Thirty-one strains of S. aureus isolates were genotypically tested by polymerase chain reaction (PCR), yielding a positive result for the icaA gene in 83.87% of the samples, 80.64% positive for icaD and 74.19% of these showed both genes. The results reinforce the importance of controlling this disease by adopting strict hygiene protocols and disinfection of equipment used for milking and the prescription of appropriate therapies to clinical and subclinical cases of mastitis, as well as evaluating not only the susceptibility profile of isolates but also the biofilm formation capacity.

Biological Evaluation of Selected 1,2,3-triazole Derivatives as Antibacterial and Antibiofilm Agents

2020 ◽  
Vol 20 (24) ◽  
pp. 2186-2191
Author(s):  
Lialyz Soares Pereira André ◽  
Renata Freire Alves Pereira ◽  
Felipe Ramos Pinheiro ◽  
Aislan Cristina Rheder Fagundes Pascoal ◽  
Vitor Francisco Ferreira ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Antimicrobial Agents ◽  
Public Health Problem ◽  
Biological Evaluation ◽  
Major Public Health Problem ◽  
Community Environments ◽  
Scanning Electron

Background: Resistance to antimicrobial agents is a major public health problem, being Staphylococcus aureus prevalent in infections in hospital and community environments and, admittedly, related to biofilm formation in biotic and abiotic surfaces. Biofilms form a complex and structured community of microorganisms surrounded by an extracellular matrix adhering to each other and to a surface that gives them even more protection from and resistance against the action of antimicrobial agents, as well as against host defenses. Methods: Aiming to control and solve these problems, our study sought to evaluate the action of 1,2,3- triazoles against a Staphylococcus aureus isolate in planktonic and in the biofilm form, evaluating the activity of this triazole through Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) tests. We have also performed cytotoxic evaluation and Scanning Electron Microscopy (SEM) of the biofilms under the treatment of the compound. The 1,2,3-triazole DAN 49 showed bacteriostatic and bactericidal activity (MIC and MBC 128 μg/mL). In addition, its presence interfered with the biofilm formation stage (1/2 MIC, p <0.000001) and demonstrated an effect on young preformed biofilm (2 MICs, p <0.05). Results: Scanning Electron Microscopy images showed a reduction in the cell population and the appearance of deformations on the surface of some bacteria in the biofilm under treatment with the compound. Conclusion: Therefore, it was possible to conclude the promising anti-biofilm potential of 1,2,3-triazole, demonstrating the importance of the synthesis of new compounds with biological activity.

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 Staphylococcal slime layers and biofilm from different origins

2019 ◽  
Vol 49 (5) ◽  
Author(s):  
Rodrigo Casquero Cunha ◽  
Michelle Dias Hornes da Rosa ◽  
Cleomar da Silva ◽  
Francisco Denis Souza Santos ◽  
Fábio Pereira Leivas Leite
Keyword(s):  
Biofilm Formation ◽  
Pathogenic Bacteria ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Bovine Mastitis ◽  
Diffusion Method ◽  
Slime Layer ◽  
Gentamicin Resistance ◽  
Different Origins ◽  
Staphylococcus Spp

ABSTRACT: The genus Staphylococcus comprises some of the most important pathogenic bacteria for both humans and animals. It is responsible for bovine mastitis and canine otitis, besides being present in the microbiota of animals and as a contaminant in food. Its pathogenesis is related to the formation of capsule and biofilm, which contribute to its infectivity. The objective of this study was to observe the production of slime layer and formation of biofilm, which are related to the resistance to antimicrobial agents and presence of icaA and icaD genes, in 41 isolates of Staphylococcus spp. from different origins, provided by the Universidade Federal de Pelotas (UFPEL), Laboratório Regional de Diagnóstico (LRD). Strains of Staphylococcus spp. were cultivated in Congo red agar for capsule detection. Biofilm formation was detected using the 96-well microplate testing. Antimicrobial susceptibility testing was performed using the plate diffusion method. Part of the analyzed samples produced slime layer (36.6%) and formed biofilm (17.1%). However, six of those that formed biofilms were susceptible to the eight antibiotics tested in the antibiogram. In tests to determine the minimum bactericidal and inhibitory concentrations, gentamicin resistance of biofilm-forming strains was greater than that of non-forming strains. Ampicillin was the least effective antimicrobial drug (51%), followed by tetracycline (71%), neomycin (73%), and erythromycin (73%). Some isolates presented the icaA (6) and icaD (11) genes. Therefore, we suggested that the origin of an isolate can determine its expression of virulence factor and resistance to certain antibiotics.

scholarly journals Bap, a Staphylococcus aureus Surface Protein Involved in Biofilm Formation

2001 ◽  
Vol 183 (9) ◽  
pp. 2888-2896 ◽  
Author(s):  
Carme Cucarella ◽  
Cristina Solano ◽  
Jaione Valle ◽  
Beatriz Amorena ◽  
Íñigo Lasa ◽  
...  
Keyword(s):  
Amino Acids ◽  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Bovine Mastitis ◽  
Surface Protein ◽  
Surface Proteins ◽  
Infection Model ◽  
Mouse Infection Model ◽  
New Genes ◽  
Inert Surfaces

ABSTRACT Identification of new genes involved in biofilm formation is needed to understand the molecular basis of strain variation and the pathogenic mechanisms implicated in chronic staphylococcal infections. A biofilm-producing Staphylococcus aureus isolate was used to generate biofilm-negative transposon (Tn917) insertion mutants. Two mutants were found with a significant decrease in attachment to inert surfaces (early adherence), intercellular adhesion, and biofilm formation. The transposon was inserted at the same locus in both mutants. This locus (bap [for biofilm associated protein]) encodes a novel cell wall associated protein of 2,276 amino acids (Bap), which shows global organizational similarities to surface proteins of gram-negative (Pseudomonas aeruginosa andSalmonella enterica serovar Typhi) and gram-positive (Enteroccocus faecalis) microorganisms. Bap's core region represents 52% of the protein and consists of 13 successive nearly identical repeats, each containing 86 amino acids. bap was present in a small fraction of bovine mastitis isolates (5% of the 350S. aureus isolates tested), but it was absent from the 75 clinical human S. aureus isolates analyzed. All staphylococcal isolates harboring bap were highly adherent and strong biofilm producers. In a mouse infection modelbap was involved in pathogenesis, causing a persistent infection.

Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation

2003 ◽  
Vol 92 (1-2) ◽  
pp. 179-185 ◽  
Author(s):  
Pradeep Vasudevan ◽  
Manoj Kumar Mohan Nair ◽  
Thirunavukkarasu Annamalai ◽  
Kumar S Venkitanarayanan
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Bovine Mastitis ◽  
Genotypic Characterization ◽  
Phenotypic And Genotypic Characterization

scholarly journals Alpha-Toxin Is Required for Biofilm Formation by Staphylococcus aureus

2003 ◽  
Vol 185 (10) ◽  
pp. 3214-3217 ◽  
Author(s):  
Nicky C. Caiazza ◽  
G. A. O'Toole
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Antimicrobial Agents ◽  
Clinical Settings ◽  
Alpha Toxin ◽  
Flow Conditions ◽  
Alpha Hemolysin ◽  
Integral Role ◽  
Hemolytic Toxin ◽  
Plastic Surfaces

ABSTRACT Staphylococcus aureus is a common pathogen associated with nosocomial infections. It can persist in clinical settings and gain increased resistance to antimicrobial agents through biofilm formation. We have found that alpha-toxin, a secreted, multimeric, hemolytic toxin encoded by the hla gene, plays an integral role in biofilm formation. The hla mutant was unable to fully colonize plastic surfaces under both static and flow conditions. Based on microscopy studies, we propose that alpha-hemolysin is required for cell-to-cell interactions during biofilm formation.

scholarly journals Antibiotic susceptibility of Staphylococcus aureus with different degrees of biofilm formation

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Hyo-Jung Shin ◽  
Sungtae Yang ◽  
Yong Lim
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Antimicrobial Agents ◽  
Resistance Mechanisms ◽  
Minimal Bactericidal Concentration ◽  
Chronic Infections ◽  
Planktonic Bacteria ◽  
Growth Modes ◽  
Biofilm Bacteria ◽  
Biofilm Development

AbstractStaphylococcus aureus is one of the most common pathogens in biofilm-associated chronic infections. S. aureus living within biofilms evades the host immune response and is more resistant to antibiotics than planktonic bacteria. In this study, we generated S. aureus with low and high levels of biofilm formation using the rbf (regulator of biofilm formation) gene and performed a BioTimer assay to determine the minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of various types of antibiotics. We showed that biofilm formation by S. aureus had a greater effect on MBC than MIC, probably due to the different growth modes between planktonic and biofilm bacteria. Importantly, we found that the MBC for biofilm S. aureus was much higher than that for planktonic cells, but there was little difference in MBC between low and high levels of biofilm formation. These results suggest that once the biofilm is formed, the bactericidal activity of antibiotics is significantly reduced, regardless of the degree of S. aureus biofilm formation. We propose that S. aureus strains with varying degrees of biofilm formation may be useful for evaluating the anti-biofilm activity of antimicrobial agents and understanding antibiotic resistance mechanisms by biofilm development.

scholarly journals Virulence, agr groups, antimicrobial resistance and epidemiology of Staphylococcus aureus isolated from bovine subclinical mastitis

2021 ◽  
Vol 58 ◽  
pp. e186701
Author(s):  
Felipe Mendes Zorzi ◽  
Luiz Francisco Zafalon ◽  
Franklin Bispo Santos ◽  
Arthur Ferreira Borges ◽  
Ticiano Gomes Nascimento ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Virulence Genes ◽  
Antimicrobial Agents ◽  
Subclinical Mastitis ◽  
Southeastern Brazil ◽  
Economic Losses ◽  
Group Iii ◽  
Oxacillin Resistance ◽  
Group Ii

Fifty-two Staphylococcus aureus recovered from papillary ostium and milk samples collected from cows with subclinical mastitis and milking environments in three small dairy herds located in southeastern Brazil were subjected to PCR identification based on the thermonuclease (nuc) gene. All the strains were submitted to in vitro antimicrobial susceptibility testing, and we investigated the sequence types (STs), agr groups (I-IV), virulence genes encoding for Microbial Surface Components Recognizing Adhesive Matrix Molecules (MSCRAMMs), biofilm-associated proteins, bi-component toxins, pyrogenic toxin superantigens, and enterotoxins. Screening for oxacillin resistance (2-6 μg/ml oxacillin), beta-lactamase activity assays, and PCR for the mecA/mecC genes detected 26 methicillin-susceptible S. aureus (MSSA) and 26 mec-independent oxacillin-nonsusceptible S. aureus (MIONSA). While MSSA isolates were found to be susceptible to all antimicrobial agents tested, or only resistant to penicillin and ampicillin, MIONSA isolates were multidrug-resistant. ST126-agr group II MSSA isolates were prevalent in milk (n=14) and carried a broad set of virulence genes (clfA, clfB, eno, fnbA, fiB, icaA, icaD, lukED, hla, and hlb), as well as the ST126-agr group II MIONSA isolated from milking liners (n=1), which also carried the eta gene. ST1-agr group III MIONSA isolates (n=4) were found in papillary ostium and milk, but most MIONSA isolates (n=21), which were identified in both papillary ostium and milking liners, were agr-negative and assigned to ST126. The agr-negative and agr group III lineages showed a low potential for virulence. Studies on the characterization of bovine-associated MSSA/MIONSA are essential to reduce S. aureus mastitis to prevent economic losses in dairy production and also to monitor the zoonotic potential of these pathogens associated with invasive infections and treatment failures in healthcare.

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