Evaluation of the presence of the bap gene in Staphylococcus aureus isolates recovered from human and animals species

doi:10.1016/j.vetmic.2007.08.018

Veterinary Microbiology

Volume 127, Issues 3–4, 18 March 2008, Pages 407-411

https://doi.org/10.1016/j.vetmic.2007.08.018 Get rights and content

Abstract

The implication of biofilm in chronic bacterial infection in many species has triggered an increasing interest in the characterization of genes involved in biofilm formation. The bap gene is a newly identified gene that encodes the biofilm-associated protein, BAP, which is involved in biofilm formation in Staphylococcus aureus. So far the bap gene has only been found in a small proportion of S. aureus strains from bovine mastitis in Spain. In order to study the presence of the bap gene in S. aureus isolates obtained from other species and various locations, a collection of 262 isolates was tested by PCR, using published primers and dot-blot. The results indicated that none of the isolates carried the bap gene suggesting that the prevalence of this gene among S. aureus isolates should be very low.

Introduction

In Staphylococcus aureus (S. aureus) the implication of biofilm in chronic infections in all animal species have triggered an increasing interest in the characterization of genes involved in this biofilm formation. For example, the biofilm formation is important for virulence in mastitis (Baselga et al., 1993). A new gene (6831 nucleotides) involved in biofilm formation (bap coding for a biofilm-associated protein, Bap) was identified in a small proportion of S. aureus from bovine mastitis (Cucarella et al., 2001). The bap protein is a member of proteins playing a role in biofilm formation in many bacteria. They share common structural features as they have a high molecular weight and contain a core domain of tandem repeats. These proteins confer upon bacteria the capacity to form a biofilm and play a relevant role in bacterial infectious process. Some of these proteins are contained occasionally in mobile elements (Lasa and Penades, 2006). In S. aureus, the bap gene is carried by a putative composite transposon inserted in SaPIbov2, a mobile staphylococcal pathogenicity island. Bap orthologue genes have been found in other staphylococcal species including Staphylococcus epidermidis, Staphylococcus chromogenes, Staphylococcus xylosus, Staphylococcus simulans and Staphylococcus hyicus. However, sequence analyses of the flanking regions revealed that these orthologue bap genes of these staphylococcal species were not contained in the SaPIbov2 pathogenicity island (Tormo et al., 2005).

The aim of the present study was to investigate the presence of bap gene in various S. aureus isolates recovered from human and different animal species. To do this a published PCR method was used (Cucarella et al., 2001) and results were confirmed by dot-blot analysis.

Section snippets

S. aureus isolates used in the study

Two hundred and sixty-two S. aureus isolates associated with different diseases were recovered from various locations in France and different animal species (cows, sheep, goats, pigs, rabbits, poultry, horses, human) (Table 1). The cow's isolates were a gift from Dr. Martel (AFSSA, Lyon). Some sheep isolates were from De Buyser (AFSSA Maisons-Alfort). The goat's isolates were partly from Dr. Mercier (AFSSA Niort). The pig, poultry and rabbits isolates were from Bäyon-Auboyer (Departemental

Results

DNA extracted from 262 S. aureus isolates was tested for the presence the bap gene by PCR, using the primer pair sasp-6m and sasp-7c, as indicated in Section 2. Although the positive control strain (V329) showed a band at 971 bp, as expected, none of the tested isolates showed positive results. All isolates were also tested for the presence of the S. aureus 23S DNA by using PCR to check for DNA quality, presence of inhibitors of the PCR reactions and specificity. All isolates were found

Discussion

The gene for the biofilm-associated protein (bap gene) was not detected in the 262 S. aureus isolates of this study. These results are in agreement with previous surveys on S. aureus of human, bovine, rabbit and pig origins (Arciola et al., 2001, Vasudevan et al., 2003, Vancraeynest et al., 2004, Nitzsche et al., 2007) where the bap gene was not found in the S. aureus isolates recovered in these animal species of these studies. Our study is the first one with a wide range of S. aureus recovered

Acknowledgement

We thank Dr. José Penades (Cardenal Herrera-CEU University and Instituto Valenciano de Investigationes Agrarias, 46113 Moncada, Valencia, Spain) for helpful discussion.

References (14)

I. Lasa et al.
Bap: a family of surface proteins involved in biofilm formation
Res. Microbiol.
(2006)
S. Nitzsche et al.
Phenotypic and genotypic traits of Staphylococcus aureus strains isolated from pig carcasses
Vet. Microbiol.
(2007)
S. Planchon et al.
Formation of biofilm by Staphylococcus xylosus
Int. J. Food Microbiol.
(2006)
J.A. Straub et al.
A 23S rDNA-targeted polymerase chain reaction-based system for detection of Staphylococcus aureus in meat starter cultures and dairy products
J. Food Prot.
(1999)
D. Vancraeynest et al.
Genotypic and phenotypic screening of high and low virulence Staphylococcus aureus isolates from rabbits for biofilm formation and MSCRAMMs
Vet. Microbiol.
(2004)
P. Vasudevan et al.
Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation
Vet. Microbiol.
(2003)
E. Vautor et al.
Characterization of Staphylococcus aureus isolates recovered from dairy sheep farms (agr group, adherence, slime, resistance to antibiotics)
Small Rumin. Res.
(2007)

There are more references available in the full text version of this article.

Cited by (59)

Antimicrobial characterization of a titanium coating derived from mussel-glue and Bothrops asper snake venom for the prevention of implant-associated infections caused by Staphylococcus
2022, Electronic Journal of Biotechnology
Citation Excerpt :
In contrast to Aap, which is found in approximately 90% of S. epidermidis isolates, Bhp is encoded in approximately 15–45% of the isolates [42]. In S. aureus, Bap is even less frequent and Vautor et al. [43] found that bhp was not encoded in 262 S. aureus isolates obtained from various animal and human sources. Bap is mostly present in bovine mastitis isolates of S. aureus, and their absence from human clinical isolates is because Bap-mediated biofilm formation seems to be a system specialized for the conditions present in the mammary gland, where calcium concentration can reach the high values necessary to modulate Bap function (∼10 mM) [44].
Proliferation of bacteria, such as Staphylococcus aureus and Staphylococcus epidermidis, on orthopedic implants has been a challenge in orthopedic surgery, highlighting the problem of the increasing antibiotic resistance and the necessity to develop new antimicrobial agents. In this sense, antimicrobial peptides are promising candidates, which can be attached to titanium surfaces in order to make them safer. Mytilus mussels are characterized by adhering efficiently to a wide variety of surfaces, especially metallic ones, through adhesive proteins with a high content of dihydroxyphenylalanine, which is a post-translational modification of tyrosine.
This work refers to the synthesis of a coating based on a bifunctional peptide that combines a sequence derived from the mussel foot protein-5 and the antimicrobial peptide pEM-2, derived from Bothrops asper snake venom. The adhesive properties of this bifunctional peptide were evaluated, as well as the adhesive sequence without the pEM-2, using a Quartz Crystal Microbalance. The results showed that the presence of the antimicrobial peptide improved the adhesion; however, a loss of the bactericidal activity was observed. Even so, the adhesive sequence by itself exhibited an important antifouling activity, preventing S. aureus and S. epidermidis adhesion to titanium by 75% and 45%, respectively, although the result against S. epidermidis was not significant.
A 13-residue peptide derived from a natural and biocompatible source, like a Mytilus mussel adhesive protein, could be projected as a protective agent on titanium surfaces against S. aureus and S. epidermidis, being responsible for two thirds of the cases of orthopedic implant infection.
How to cite: Gauna A, Mercado L, Guzmán F. Antimicrobial characterization of a titanium coating derived from mussel-glue and Bothrops asper snake venom for the prevention of implant-associated infections caused by Staphylococcus. Electron J Biotechnol 2022;56. https://doi.org/10.1016/j.ejbt.2022.02.001.
Invited review: A critical appraisal of mastitis vaccines for dairy cows
2021, Journal of Dairy Science
Infections of the mammary gland remain a frequent disease of dairy ruminants that negatively affect animal welfare, milk quality, farmer serenity, and farming profitability and cause an increase in use of antimicrobials. There is a need for efficacious vaccines to alleviate the burden of mastitis in dairy farming, but this need has not been satisfactorily fulfilled despite decades of research. A careful appraisal of past and current research on mastitis vaccines reveals the peculiarities but also the commonalities among mammary gland infections associated with the major mastitis pathogens Escherichia coli, Staphylococcus aureus, Streptococcus uberis, Streptococcus agalactiae, or Streptococcus dysgalactiae. A major pitfall is that the immune mechanisms of effective protection have not been fully identified. Until now, vaccine development has been directed toward the generation of antibodies. In this review, we drew up an inventory of the main approaches used to design vaccines that aim at the major pathogens for the mammary gland, and we critically appraised the current and tentative vaccines. In particular, we sought to relate efficacy to vaccine-induced defense mechanisms to shed light on some possible reasons for current vaccine shortcomings. Based on the lessons learned from past attempts and the recent results of current research, the design of effective vaccines may take a new turn in the years to come.
Presence of biofilm and adhesin genes in Staphylococcus aureus strains taken from chronic wound infections and their genotypic and phenotypic antimicrobial sensitivity patterns
2020, Photodiagnosis and Photodynamic Therapy
The purpose of this research was to examine biofilm (icaA, icaD and bap) and adhesin (clfA, fnbA, cna) genes, and also assess the genotypic and phenotypic antimicrobial resistance patterns of Staphylococcus aureus strains taken from wound specimens in Mardin, Turkey. A total of 220 wound specimens were investigated. The biofilm forming ability and resistance pattern for eleven antimicrobial agents were investigated by conventional and multiplex PCR methods. S. aureus were taken from 112 (50.9%) of 220 wound specimens. Moreover, biofilm production was found in 79 (70.5%) of the 112 S. aureus isolates. 97 (86.6%) strains of all isolates were positive for icaA and icaD, and 15 (13.4%) for bap. The adhesin genes, cna, fnbA and clfA were detected in 98 (87.5%), 87 (77.7%), and 75 (66.9%) strains, respectively. The numbers of MSSA and MRSA bearing antimicrobial resistance genes were 19 (16.96%) and 32 (28.57%) for blaZ, 9 (8.04%) and 17 (15.18%) for tetK, 6 (5.36%) and 14 (12.5%) for ermC, 2 (1.79%) and 7 (6.25%) for tetM, 0 (0%) and 5 (4.46%) for mecA, 2 (1.79%) and 4 (3.57%) for ermA, 1 (0.89%) and 2 (1.79%) for both tetK and tetM, respectively. Our findings indicate that multiplex PCR is a suitable way for identifying biofilm and adhesin producing S. aureus. Our data also provided a country-wide oversight of the S. aureus antimicrobial resistance gene profiles for the properly therapy of patients and to control the spreading of the resistance genes.
Genotypic and phenotypic characterization of biofilm production by Staphylococcus aureus strains isolated from bovine intramammary infections in Colombian dairy farms
2019, Heliyon
The ability of Staphylococcus aureus to form biofilms is an important virulence factor because this has been associated with persistent bovine intramammary infections. Different mechanisms of biofilm formation have been described in S. aureus; however, the process has been found to be mainly driven by the ica and bap genes. The presence of the ica and bap genes, as well as the biofilm formation in vitro were evaluated in 229 S. aureus strains isolated from bovine milk collected from different regions of Department of Antioquia, Colombia. Three different genotypes grouped into three separate clusters were identified from in vitro assays. Genotype 1 (ica positive and bap negative) was the most prevalent (78.17%), followed by genotype 2 (ica and bap positive) (12.66%) and genotype 0 (ica and bap negative) (9.17%). Biofilm formation was observed in 81.26% of the strains from which 100% of genotype 2 isolates showed biofilm formation. The biofilms formed by genotype 2 isolates were also found to have the highest optical density (>2.4). These results showed that most of the S. aureus strains were capable of biofilm formation, suggesting the virulence potential particularly in bap-positive strains.
The Role of Functional Amyloids in Bacterial Virulence
2018, Journal of Molecular Biology
Amyloid fibrils are best known as a product of human and animal protein misfolding disorders, where amyloid formation is associated with cytotoxicity and disease. It is now evident that for some proteins, the amyloid state constitutes the native structure and serves a functional role. These functional amyloids are proving widespread in bacteria and fungi, fulfilling diverse functions as structural components in biofilms or spore coats, as toxins and surface-active fibers, as epigenetic material, peptide reservoirs or adhesins mediating binding to and internalization into host cells. In this review, we will focus on the role of functional amyloids in bacterial pathogenesis. The role of functional amyloids as virulence factor is diverse but mostly indirect. Nevertheless, functional amyloid pathways deserve consideration for the acute and long-term effects of the infectious disease process and may form valid antimicrobial targets.
Role of BP*C@AgNPs in Bap-dependent multicellular behavior of clinically important methicillin-resistant Staphylococcus aureus (MRSA) biofilm adherence: A key virulence study
2018, Microbial Pathogenesis
Bacterial adhesion is a threshold event in the formation of biofilms which leads to serious bacterial diseases. This shows that the underlining the problem is interesting and need to solve the problem of biofilm-related complications. To support this, in the present study, we first time initiated to understand the role of methicillin-resistant Staphylococcus aureus (MRSA) biofilm using previously developed benzodioxane midst piperazine decorated chitosan silver nanoparticles (BP*C@AgNPs). The BP*C@AgNPs studied for antimicrobial, anti-biofilm, biofilm adherence inhibition, the role of ions in biofilm, and an antibiotic cocktail in the treatment of biofilm was assessed. The results showed that, the significant biocidal role of BP*C@AgNPs in controlling the MRSA biofilm and interaction of biofilm protein to calcium ions were significantly decreased. This confirms that calcium ion involved in the biofilm formation and for the treatment of BP*C@AgNPs, cocktail of enzyme and antibiotic have the promising therapeutic value was observed. In future the locking of biofilm protein and its expression in presence of calcium ion was interesting, and greater application related to biofilm infection was warrantable.

View all citing articles on Scopus

View full text

Article preview

Veterinary Microbiology

Abstract

Introduction

Section snippets

S. aureus isolates used in the study

Results

Discussion

Acknowledgement

References (14)

Bap: a family of surface proteins involved in biofilm formation

Res. Microbiol.

Phenotypic and genotypic traits of Staphylococcus aureus strains isolated from pig carcasses

Vet. Microbiol.

Formation of biofilm by Staphylococcus xylosus

Int. J. Food Microbiol.

A 23S rDNA-targeted polymerase chain reaction-based system for detection of Staphylococcus aureus in meat starter cultures and dairy products

J. Food Prot.

Genotypic and phenotypic screening of high and low virulence Staphylococcus aureus isolates from rabbits for biofilm formation and MSCRAMMs

Vet. Microbiol.

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

Vet. Microbiol.

Characterization of Staphylococcus aureus isolates recovered from dairy sheep farms (agr group, adherence, slime, resistance to antibiotics)

Small Rumin. Res.

Cited by (59)

Antimicrobial characterization of a titanium coating derived from mussel-glue and Bothrops asper snake venom for the prevention of implant-associated infections caused by Staphylococcus

Invited review: A critical appraisal of mastitis vaccines for dairy cows

Presence of biofilm and adhesin genes in Staphylococcus aureus strains taken from chronic wound infections and their genotypic and phenotypic antimicrobial sensitivity patterns

Genotypic and phenotypic characterization of biofilm production by Staphylococcus aureus strains isolated from bovine intramammary infections in Colombian dairy farms

The Role of Functional Amyloids in Bacterial Virulence

Role of BP*C@AgNPs in Bap-dependent multicellular behavior of clinically important methicillin-resistant Staphylococcus aureus (MRSA) biofilm adherence: A key virulence study

Short communicationEvaluation of the presence of the bap gene in Staphylococcus aureus isolates recovered from human and animals species

Abstract

Introduction

Section snippets

S. aureus isolates used in the study

Results

Discussion

Acknowledgement

Res. Microbiol.

Vet. Microbiol.

Int. J. Food Microbiol.

J. Food Prot.

Vet. Microbiol.

Vet. Microbiol.

Small Rumin. Res.

Short communication
Evaluation of the presence of the bap gene in Staphylococcus aureus isolates recovered from human and animals species