Study on Biofilm Formation and Multiplex PCR of icaABCD Operon in Staphylococcus capitis

2021 ◽  
pp. 127-138
Author(s):  
Aziyah Abdul-Aziz ◽  
Mohamad Faiz Foong Abdullah ◽  
Sharifah Aminah Syed Mohamad
Keyword(s):  
Multiplex Pcr ◽  
Biofilm Formation ◽  
Staphylococcus Capitis

scholarly journals Biofilm Formation and Multiplex PCR detection of icaABCD Operon in Staphylococcus capitis

2021 ◽  
pp. 223-234
Author(s):  
Aziyah Abdul-Aziz ◽  
Mohamad Faiz Foong Abdullah ◽  
Sharifah Aminah Syed Mohamad
Keyword(s):  
Multiplex Pcr ◽  
Dna Sequences ◽  
Biofilm Formation ◽  
Opportunistic Infections ◽  
Opportunistic Pathogen ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Coagulase Negative Staphylococcus ◽  
Microtitre Plate ◽  
Staphylococcus Capitis

Aims: The ability to form biofilm is a major virulence factor in the virulence of the Coagulase negative Staphylococcus (CoNS) group of bacteria. Being the most predominant member of CoNS, the ability of S. epidermidis in causing biofilm-associated infections has been well established. On the other hand, S. capitis and has always been regard as a non-pathogenic species although recently it was found to be responsible in a variety of infections. Hence, this study aimed to determine the biofilm formation capabilities and the presence of icaABCD genes in clinical isolates of S. capitis, which have emerged as an important opportunistic pathogen in clinical settings. Methodology: S. capitis was isolated and identified from 17 out of 200 clinical samples. Biofilm formation assay was performed quantitatively using a microtitre plate method. Mulitplex PCR primers for icaABCD genes were designed from DNA sequences coding for the icaA, B, C, and D structural genes of S capitis JF930147.1 which was compared together with five other species of Staphylococcus. Amplification of the icaABCD genes was performed using the designated primers. Results: From the 17 strains of S. capitis clinical isolates, 14 were identified as S. capitis subsp capitis while the remaining three were identified as S. capitis subsp ureolyticus. Except for two of the S. capitis subsp capitis isolates, the remaining strains were able to form biofilm, with majority of them were strong biofilm formers. Multiplex PCR was successful in amplifying the four icaABCD genes which was demonstrated in all the S. capitis isolates, including the two non-biofilm forming isolates. Conclusion: Majority of the S. capitis isolates were able to form biofilm phenotypically suggesting the possibility in causing opportunistic infections through indwelling medical devices. Multiplex PCR however was able to detect the presence of the icaABCD genes in all the S. capitis isolates. This suggests that the biofilm assessment on microtitre plate is not a definitive tool in determining the production of polysaccharide intercellular adhesion (PIA) but the production of the icaABCD genes could be a better assessment in determining biofilm production in Staphylococcus.

Biofilm Formation and Associated Genes in Listeria Monocytogenes

2017 ◽  
Vol 12 (3) ◽  
Author(s):  
S. R. Warke ◽  
V. C. Ingle ◽  
N. V. Kurkure ◽  
P. A. Tembhurne ◽  
Minakshi Prasad ◽  
...  
Keyword(s):  
Public Health ◽  
Listeria Monocytogenes ◽  
Multiplex Pcr ◽  
Food Processing ◽  
Biofilm Formation ◽  
Milk Samples ◽  
Biofilm Production ◽  
Food Borne ◽  
Serious Infections ◽  
Microtiter Assay

Listeria monocytogenes, an opportunistic food borne pathogen can cause serious infections in immunocompromised individuals. L. monocytogenes is capable of producing biofilm on the surface of food processing lines and instruments.The biofilm transfers contamination to food products and impose risk to public health. In the present study biofilm producing ability of L. monocytogenes isolates were investigated phenotypically and genotypically by microtiter assay and multiplex PCR, respectively. Out of 38 L. monocytogenes isolates 14 were recovered from animal clinical cases, 12 bovine environment and 12 from milk samples. A total of 3 (21.42%) clinical, 2 (16.66%) environment and 3 (25%) milk samples respectively, revealed biofilm production in microtiter assay. Cumulative results showed that 23 (60.52%) out of 38 strains of L. monocytogenes were positive for luxS and flaA gene and 1 (2.63%) was positive only for the flaA gene.

Characterization of biofilm producing methicillin resistant coagulase negative Staphylococci from India

2021 ◽  
Keyword(s):  
Biofilm Formation ◽  
Resistance Rate ◽  
Coagulase Negative Staphylococci ◽  
Methicillin Resistant ◽  
Antimicrobial Susceptibility Pattern ◽  
Healthcare Settings ◽  
Staphylococcus Capitis ◽  
Staphylococcus Epidermis ◽  
Staphylococcus Hominis

Abstract Methicillin-resistant coagulase-negative staphylococci (MR-CoNS) cause infectious diseases due to their potential to form biofilm and further colonization in hospital materials. This study evaluated the antibiotic susceptible phenotypes, biofilm-producing ability, and biofilm-associated genes (mecA, icaAD, bap, cna, and fnbA). Biofilm formation was detected through Congo red agar (CRA) method and MTP method. The presence of biofilm and associated genes in MR-CoNS were detected by PCR. A total of 310 (55.95%) isolates produced the biofilm. Among these isolates, Staphylococcus haemolyticus (34.83%), Staphylococcus epidermis (31.93%), Staphylococcus capitis (16.77%), Staphylococcus cohnii (10.96%), and Staphylococcus hominis (5.48%) were identified. The antimicrobial susceptibility pattern of CoNS isolates indicated resistance to cefoxitin (100%), erythromycin (94.8%), ciprofloxacin (66.7%), sulfamethoxazole/trimethoprim (66.7%), gentamicin (66.12%), and clindamycin (62.9%). Resistance rate to mupirocin was 48.5% in S. epidermidis and 38.9% in S. haemolyticus isolates. All isolates were sensitive to vancomycin and linezolid. The prevalence rates of icaAD, bap, fnbA, and cna were 18.06%, 12.5%, 47.4%, and 27.4%, respectively. icaAD and bap genes were detected in 18.06% and 12.5% of MR-CoNS isolates. fnbA and cna genes were detected in 47.41% and 27.41% of MRCoNS isolates. icaAD positive strains exhibited a significant increase in the biofilm formation compared with those that lacked icaAD (0.86 (0.42, 1.39) versus 0.36 (0.14, 0.75), respectively; P < 0.001). In conclusion, the majority of MR-CoNS isolates were biofilm producers, and S. capitis, which possessed icaAD genes, ranked as the great biofilm producer than other Staphylococcus. The study’s findings are important to form a strategy to control biofilm formation as an alternative strategy to counter the spread of MR-CoNS in healthcare settings.

Development of a Multiplex PCR Assay for Detection ofPseudomonas fluorescenswith Biofilm Formation Ability

2017 ◽  
Vol 82 (10) ◽  
pp. 2337-2342 ◽  
Author(s):  
Yu Xu ◽  
Wanyi Chen ◽  
Chunping You ◽  
Zhenmin Liu
Keyword(s):  
Multiplex Pcr ◽  
Biofilm Formation ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

scholarly journals Testing Laser-Structured Antimicrobial Surfaces Under Space Conditions: The Design of the ISS Experiment BIOFILMS

2022 ◽  
Vol 2 ◽  
Author(s):  
Katharina Siems ◽  
Daniel W. Müller ◽  
Laurens Maertens ◽  
Aisha Ahmed ◽  
Rob Van Houdt ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Space Station ◽  
Health And Safety ◽  
Model Organisms ◽  
Antimicrobial Surfaces ◽  
Microbial Biofilms ◽  
Staphylococcus Capitis ◽  
Antimicrobial Materials ◽  
Direct Laser ◽  
Inert Surfaces

Maintaining crew health and safety are essential goals for long-term human missions to space. Attaining these goals requires the development of methods and materials for sustaining the crew’s health and safety. Paramount is microbiological monitoring and contamination reduction. Microbial biofilms are of special concern, because they can cause damage to spaceflight equipment and are difficult to eliminate due to their increased resistance to antibiotics and disinfectants. The introduction of antimicrobial surfaces for medical, pharmaceutical and industrial purposes has shown a unique potential for reducing and preventing biofilm formation. This article describes the development process of ESA’s BIOFILMS experiment, that will evaluate biofilm formation on various antimicrobial surfaces under spaceflight conditions. These surfaces will be composed of different metals with and without specified surface texture modifications. Staphylococcus capitis subsp. capitis, Cupriavidus metallidurans and Acinetobacter radioresistens are biofilm forming organisms that have been chosen as model organisms. The BIOFILMS experiment will study the biofilm formation potential of these organisms in microgravity on the International Space Station on inert surfaces (stainless steel AISI 304) as well as antimicrobial active copper (Cu) based metals that have undergone specific surface modification by Ultrashort Pulsed Direct Laser Interference Patterning (USP-DLIP). Data collected in 1 x g has shown that these surface modifications enhance the antimicrobial activity of Cu based metals. In the scope of this, the interaction between the surfaces and bacteria, which is highly determined by topography and surface chemistry, will be investigated. The data generated will be indispensable for the future selection of antimicrobial materials in support of human- and robotic-associated activities in space exploration.

Biofilm formation by Staphylococcus capitis strains isolated from contaminated platelet concentrates

2013 ◽  
Vol 62 (7) ◽  
pp. 1051-1059 ◽  
Author(s):  
Valerie S. Greco-Stewart ◽  
Hamza Ali ◽  
Dilini Kumaran ◽  
M. Kalab ◽  
Ineke G. H. Rood ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Extracellular Polysaccharide ◽  
Regulatory Elements ◽  
Proteinase K ◽  
Blood Collection ◽  
Platelet Concentrates ◽  
Sodium Metaperiodate ◽  
Staphylococcus Capitis ◽  
Skin Disinfection ◽  
Skin Flora

Bacterial contamination of platelet concentrates (PCs) poses the greatest infectious risk in modern transfusion medicine despite the implementation of measures such as improved skin disinfection and first aliquot diversion. The majority of PC contaminants are commensal skin flora introduced by venipuncture at the time of blood collection. The predominant organisms are Gram-positive coagulase-negative staphylococci such as Staphylococcus capitis. This bacterium has been implicated in numerous instances of infection and sepsis, likely for its ability to form surface-associated communities of micro-organisms encased in extracellular materials, known as biofilms. In the present study, five strains of S. capitis isolated from contaminated PCs were assessed for their ability to produce extracellular polysaccharide (slime), a canonical indicator of biofilm-formation ability, on Congo red agar plates. Biofilm formation was evaluated in both glucose-enriched trypticase soy broth (TSBg) and in PCs by using a crystal violet staining assay. The chemical nature of the biofilms was evaluated by disruption assays using sodium metaperiodate and proteinase K. In addition, biofilm architecture was observed by scanning electron microscopy. The presence of the biofilm-associated icaR and icaADBC genes was also examined by PCR. While only two out of the five S. capitis strains formed biofilms in TSBg, all strains formed biofilms in PCs. The ability of strains to produce extracellular polysaccharide and their possession of wild-type ica genes were not exclusive predictors of biofilm formation in TSBg or PCs; different profiles of biofilm markers were observed among isolates. This is likely due to the proteinaceous composition of the S. capitis biofilm matrix. Interestingly, an ica-negative, non-slime-producing isolate was capable of biofilm formation in PCs. Together, these data indicate that the platelet storage environment stimulates biofilm formation in S. capitis in the absence of extracellular polysaccharide production and that multiple bacterial factors and regulatory elements are likely involved in biofilm formation in this milieu.

Biofilm formation by Staphylococcus capitis strains isolated from contaminated platelet concentrates

2013 ◽  
Vol 62 (12) ◽  
pp. 1917-1917
Author(s):  
Valerie S. Greco-Stewart ◽  
Hamza Ali ◽  
Dilini Kumaran ◽  
M. Kalab ◽  
Ineke G. H. Rood ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Platelet Concentrates ◽  
Staphylococcus Capitis

scholarly journals Biofilm-Forming Ability of Microbacterium lacticum and Staphylococcus capitis Considering Physicochemical and Topographical Surface Properties

Foods ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 611
Author(s):  
Elena Zand ◽  
Hedwig Pfanner ◽  
Konrad J. Domig ◽  
Gerhard Sinn ◽  
Marija Zunabovic-Pichler ◽  
...  
Keyword(s):  
Late Stage ◽  
Biofilm Formation ◽  
Extracellular Polymeric Substances ◽  
Surface Coverage ◽  
304 Stainless Steel ◽  
Suitable Model ◽  
Strong Correlations ◽  
Staphylococcus Capitis ◽  
Total Surface Free Energy ◽  
Biofilm Structure

Biofilm characteristics of Microbacterium lacticum D84 (M. lacticum) and Staphylococcus capitis subsp. capitis (S. capitis) on polytetrafluoroethylene and AISI-304 stainless steel at early- (24, 48 h) and late-stage (144, 192 h) biofilm formation were investigated. M. lacticum biofilm structure was more developed compared to S. capitis, representing vastly mature biofilms with a strongly developed amorphous matrix, possibly extracellular polymeric substances (EPSs), at late-stage biofilm formation. S. capitis showed faster growth behavior but still resulted in a relatively flat biofilm structure. Strong correlations were found between several roughness parameters and S. capitis surface coverage (r ≥ 0.98), and between total surface free energy (γs) and S. capitis surface coverage (r = 0.89), while M. lacticum remained mostly unaffected. The pronounced ubiquitous biofilm characteristics make M. lacticum D84 a suitable model for biofilm research. Studying biofilm formation of these bacteria may help one understand bacterial adhesion on interfaces and hence reduce biofilm formation in the food industry.

scholarly journals Evaluation of the occurrence of genetic determinants of multi-drug resistance among Acinetobacter baumannii strains

2021 ◽  
pp. 15-23
Keyword(s):  
Drug Resistance ◽  
Acinetobacter Baumannii ◽  
Multiplex Pcr ◽  
Biofilm Formation ◽  
Insertion Sequence ◽  
Efflux Pump ◽  
Multi Drug Resistance ◽  
Genetic Determinants ◽  
Rapd Pcr ◽  
Pcr Method

Introduction: The aim of the study was the analysis of occurrence of genetic determinants of multi-drug resistance and the assessment of genetic relationship among Acinetobacter baumannii strains. Methods: Multiplex-PCR method was performed in order to: (1) confirm the phenotypic identification and (2) detect the presence of CHDL oxacillinases in the group of thirty A.baumannii strains. Further PCR studies included the analysis of the occurrence of genetic determinants associated with efflux pump, insertion sequence and biofilm formation. The relationship between bacterial strains was assayed using 6 primers in RAPD-PCR method. Results: Detection of the blaOXA-51-like gene confirmed that the strains belong to the A. baumannii species. In the multiplex-PCR, the presence of the blaOXA-23-like and blaOXA-40-like genes was detected in 3 (10%) and 27 (90%) isolates, respectively. Moreover, some strains showed the coexistence of the blaOXA-51-like and blaOXA-23-like genes (10%, n=3) or blaOXA-51-like and blaOXA-40-like (90%, n=27). In the group of analysed strains the presence of the efflux pump gene (adeA) and the insertion sequence ISAba1 were demonstrated in all tested isolates. Biofilm-related genes (abaI, csuE) were found in 100% and 97% (n=29) tested strains adequately. The RAPD-PCR studies revealed the presence of 10 unrelated genotypes. Conclusions: The obtained results suggest that the phenomenon of multi-drug resistance in the studied A. baumannii strains could be attributed to the occurrence of CHDL oxacillinases, AdeABC efflux pump, insertion sequence ISAba1 and the biofilm formation.

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