icaA Gene of Staphylococcus aureus Responds to NaCl, Leading to Increased Biofilm Formation

2018 ◽  
Vol 81 (3) ◽  
pp. 412-416 ◽  
Author(s):  
Soomin Lee ◽  
Sejeong Kim ◽  
Heeyoung Lee ◽  
Jimyeong Ha ◽  
Jeeyeon Lee ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Deletion Mutant ◽  
Microtiter Plate ◽  
Temperature Sensitive ◽  
Wild Type ◽  
Microtiter Plate Assay ◽  
Nacl Concentration ◽  
Icaa Gene

ABSTRACT The objective of this study was to elucidate the role of the icaA gene in biofilm formation of Staphylococcus aureus exposed to NaCl. The icaA-deletion mutant of S. aureus ATCC 13565 was constructed with the temperature-sensitive plasmid pIMAY. Microtiter plate assays were performed to confirm biofilm formation for both the wild type and the mutant at 0% (control), 2, 4, and 6% NaCl. The microtiter plate assay revealed that biofilm formation by the wild type increased (P < 0.05) as NaCl concentration increased, but biofilm formation of the mutant was not affected by NaCl concentration. Biofilm formation by the mutant was lower (P < 0.05) than that by the wild type. These results indicate that icaA plays an important role in biofilm formation by S. aureus when the pathogen is exposed to NaCl.

scholarly journals Biofilm Formation by Psychrobacter arcticus and the Role of a Large Adhesin in Attachment to Surfaces

2013 ◽  
Vol 79 (13) ◽  
pp. 3967-3973 ◽  
Author(s):  
Shannon M. Hinsa-Leasure ◽  
Cassandra Koid ◽  
James M. Tiedje ◽  
Janna N. Schultzhaus
Keyword(s):  
Biofilm Formation ◽  
Time Course ◽  
Microtiter Plate ◽  
Bacterial Attachment ◽  
Wild Type ◽  
Microtiter Plate Assay ◽  
Content Type ◽  
Reading Frame ◽  
Transposon Mutants

ABSTRACTPsychrobacter arcticusstrain 273-4, an isolate from a Siberian permafrost core, is capable of forming biofilms when grown in minimal medium under laboratory conditions. Biofilms form at 4 to 22°C when acetate is supplied as the lone carbon source and with 1 to 7% sea salt.P. arcticusis also capable of colonizing quartz sand. Transposon mutagenesis identified a gene important for biofilm formation byP. arcticus. Four transposon mutants were mapped to a 20.1-kbp gene, which is predicted to encode a protein of 6,715 amino acids (Psyc_1601). We refer to this open reading frame ascat1, for cold attachment gene 1. Thecat1mutants are unable to form biofilms at levels equivalent to that of the wild type, and there is no impact on the planktonic growth characteristics of the strains, indicating a specific role in biofilm formation. Through time course studies of the static microtiter plate assay, we determined thatcat1mutants are unable to form biofilms equivalent to that of the wild type under all conditions tested. In flow cell experiments,cat1mutants initially are unable to attach to the surface. Over time, however, they form microcolonies, an architecture very different from that produced by wild-type biofilms. Our results demonstrate that Cat1 is involved in the initial stages of bacterial attachment to surfaces.

Inhibitory Effects of Citral, Cinnamaldehyde, and Tea Polyphenols on Mixed Biofilm Formation by Foodborne Staphylococcus aureus and Salmonella Enteritidis

2014 ◽  
Vol 77 (6) ◽  
pp. 927-933 ◽  
Author(s):  
HONGMEI ZHANG ◽  
WENYUAN ZHOU ◽  
WENYAN ZHANG ◽  
ANLIN YANG ◽  
YANLAN LIU ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Salmonella Enteritidis ◽  
Food Additives ◽  
Microtiter Plate ◽  
Tea Polyphenols ◽  
Natural Food ◽  
Inhibitory Effects ◽  
Microtiter Plate Assay ◽  
Communication Signal

Biofilms are significant hazards in the food industry. In this study, we investigated the effects of food additive such as citral, cinnamaldehyde, and tea polyphenols on mixed biofilm formation by foodborne Staphylococcus aureus and Salmonella serotype Enteritidis. The adhesion rates of mixed strains in sub-MIC of additives were determined by a microtiter plate assay and bacterial communication signal autoinducer 2 (AI-2) production via a bioluminescence reporter Vibrio harveyi BB170. The structure of mixed biofilm was analyzed using scanning electron microscopy. The effect of the disinfectants hydrogen peroxide, sodium hypochlorite, and peracetic acid was tested on the mixed biofilm. Our results demonstrated that citral, cinnamaldehyde, and tea polyphenols were able to significantly inhibit mixed biofilm formation, while citral could reduce the synthesis of AI-2. Conversely, we observed a significant increase in AI-2 mediated by cinnamaldehyde. Tea polyphenols at lower concentrations induced AI-2 synthesis; however, AI-2 synthesis was significantly inhibited at higher concentrations (300 μg/ml). Food additives inhibited the adhesion of mixed bacteria on stainless steel chips and increased the sensitivity of the mixed biofilm to disinfectants. In conclusion, citral, cinnamaldehyde, and tea polyphenols had strong inhibitory effects on mixed biofilm formation and also enhanced the effect of disinfectant on mixed biofilm formation. This study provides a scientific basis for the application of natural food additives to control biofilm formation of foodborne bacteria.

Inhibitory effect of probiotic yeast Saccharomyces cerevisiae on biofilm formation and expression of α-hemolysin and enterotoxin A genes of Staphylococcus aureus

2019 ◽  
Author(s):  
Navid Saidi ◽  
Parviz Owlia ◽  
Seyed Mahmoud Amin Marashi ◽  
Horieh Saderi
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Food Poisoning ◽  
Microtiter Plate ◽  
Inhibitory Effect ◽  
Yeast Saccharomyces Cerevisiae ◽  
Microtiter Plate Assay ◽  
Probiotic Yeast ◽  
Mrsa Strains

Background and Objectives: Staphylococcus aureus, as an opportunistic pathogen, is the cause of a variety of diseases from mild skin infections to severe invasive infections and food poisoning. Increasing antibiotic resistance in S. aureus isolates has become a major threat to public health. The use of compounds produced by probiotics can be a solution to this problem. Thus, the purpose of this study was to investigate the effect of Saccharomyces cerevisiae on some virulence factors (biofilm, α-hemolysin, and enterotoxin A) of S. aureus. Materials and Methods: Supernatant and lysate extracts were prepared from S. cerevisiae S3 culture. Sub-MIC concen- trations of both extracts were separately applied to S. aureus ATCC 29213 (methicillin-sensitive S. aureus; MSSA) and S. aureus ATCC 33591 (methicillin-resistant S. aureus; MRSA) strains. Biofilm formation of these strains was measured by microtiter plate assay and expression level of α-hemolysin and enterotoxin A genes (hla and sea, respectively) using real-time PCR technique. Results: The supernatant extract has reduced both biofilm formation and expression of sea and hla genes, while lysate ex- tract had only anti-biofilm effects. The MRSA strain showed more susceptibility to yeast extracts than MSSA strain in all tests. Conclusion: The present study exhibited favorable antagonistic effects of S. cerevisiae S3, as a probiotic yeast, on MSSA and MRSA strains. Based on the findings of this study, the compounds produced by this yeast can be used to control S. aureus infections; however, further similar studies should be conducted to confirm the findings of the present study.

Function of pYV Plasmid on Biofilm Formation of Yersinia enterocolitica ERL032123 in the Presence of Ca2+

2019 ◽  
Vol 82 (10) ◽  
pp. 1683-1687
Author(s):  
HAORAN WANG ◽  
JON PALMER ◽  
STEVE FLINT
Keyword(s):  
Yersinia Enterocolitica ◽  
Calcium Ions ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Virulence Plasmid ◽  
Wild Type ◽  
Microtiter Plate Assay ◽  
Enterocolitica Strain ◽  
Plate Assay ◽  
Scanning Electron

ABSTRACT The effect of the virulence plasmid pYV and calcium ions on biofilm of Yersinia enterocolitica biofilm formation was determined using a microtiter plate assay. Loss of the pYV plasmid prevented biofilm formation and the presence of Ca2+ enhanced biofilm formation in cultures containing the pYV plasmid. Scanning electron microscopy supported the result from the microtiter plate assay showing that in the presence of Ca2+, the wild-type Y. enterocolitica strain formed a strong biofilm on a polycarbonate surface. The results implied that Ca2+ promotes Y. enterocolitica biofilm formation through the function of the pYV plasmid.

scholarly journals Salmonella enterica Serovar Typhimurium Swarming Mutants with Altered Biofilm-Forming Abilities: Surfactin Inhibits Biofilm Formation

2001 ◽  
Vol 183 (20) ◽  
pp. 5848-5854 ◽  
Author(s):  
Joe Robert Mireles ◽  
Adam Toguchi ◽  
Rasika M. Harshey
Keyword(s):  
Polyvinyl Chloride ◽  
Salmonella Enterica ◽  
Biofilm Formation ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Microtiter Plate ◽  
Wild Type ◽  
Swarming Motility ◽  
Microtiter Plate Assay ◽  
Serovar Typhimurium ◽  
Transposon Mutants

ABSTRACT Swarming motility plays an important role in surface colonization by several flagellated bacteria. Swarmer cells are specially adapted to rapidly translocate over agar surfaces by virtue of their more numerous flagella, longer cell length, and encasement of slime. The external slime provides the milieu for motility and likely harbors swarming signals. We recently reported the isolation of swarming-defective transposon mutants of Salmonella enterica serovar Typhimurium, a large majority of which were defective in lipopolysaccharide (LPS) synthesis. Here, we have examined the biofilm-forming abilities of the swarming mutants using a microtiter plate assay. A whole spectrum of efficiencies were observed, with LPS mutants being generally more proficient than wild-type organisms in biofilm formation. Since we have postulated that O-antigen may serve a surfactant function during swarming, we tested the effect of the biosurfactant surfactin on biofilm formation. We report that surfactin inhibits biofilm formation of wild-type S. enterica grown either in polyvinyl chloride microtiter wells or in urethral catheters. Other bio- and chemical surfactants tested had similar effects.

scholarly journals Cellobiose-Specific Phosphotransferase System of Klebsiella pneumoniae and Its Importance in Biofilm Formation and Virulence

2012 ◽  
Vol 80 (7) ◽  
pp. 2464-2472 ◽  
Author(s):  
Meng-Chuan Wu ◽  
Ying-Chun Chen ◽  
Tzu-Lung Lin ◽  
Pei-Fang Hsieh ◽  
Jin-Town Wang
Keyword(s):  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Type Strain ◽  
Minimal Medium ◽  
Wild Type Strain ◽  
Microtiter Plate ◽  
Wild Type ◽  
Phosphotransferase System ◽  
Microtiter Plate Assay ◽  
Content Type

ABSTRACTKlebsiella pneumoniaeis a Gram-negative bacillus belonging to the familyEnterobacteriaceae. In the past 20 years,K. pneumoniaehas become the predominant pathogen causing community-acquired pyogenic liver abscess (PLA). The formation of biofilm facilitates bacterial colonization and has been implicated in reduced susceptibility to the host immune response. To investigate genes related to biofilm formation in a PLA-associatedK. pneumoniaestrain, a transposon mutant library was screened by microtiter plate assay to identify isolates impaired for biofilm formation. One of the mutants was disrupted incelB, encoding the putative cellobiose-specific subunit IIC of enzyme II (EIIC) of a carbohydrate phosphotransferase system (PTS). This transmembrane protein is responsible for recognizing and binding specific sugars and transporting them across the cell membrane into the cytoplasm. Deletion and chromosomal complementation ofcelBconfirmed, by microtiter plate and slide culture assays, thatcelBwas indeed responsible for biofilm formation. Cellobiose-specific PTS activities of deletion mutants grown in LB broth and 0.005% cellobiose minimal medium were markedly lower than that of the wild-type strain grown under the same conditions, thereby confirming the involvement ofcelBin cellobiose transport. In 0.005% cellobiose minimal medium, thecelBmutant showed a delay in growth compared to the wild-type strain. In a mouse model of intragastric infection, deletion of thecelBgene increased the survival rate from 12.5% to 87.5%, which suggests that thecelBdeletion mutant also exhibited reduced virulence. Thus, thecelBlocus ofK. pneumoniae may contribute to biofilm formation and virulence through the metabolism of cellobiose.

scholarly journals Differential Roles of Poly-N-Acetylglucosamine Surface Polysaccharide and Extracellular DNA in Staphylococcus aureus and Staphylococcus epidermidis Biofilms

2007 ◽  
Vol 74 (2) ◽  
pp. 470-476 ◽  
Author(s):  
Era A. Izano ◽  
Matthew A. Amarante ◽  
William B. Kher ◽  
Jeffrey B. Kaplan
Keyword(s):  
Staphylococcus Aureus ◽  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Cetylpyridinium Chloride ◽  
Microtiter Plate ◽  
Dnase I ◽  
Extracellular Dna ◽  
Human Pathogens ◽  
Microtiter Plate Assay ◽  
Surface Polysaccharide

ABSTRACT Staphylococcus aureus and Staphylococcus epidermidis are major human pathogens of increasing importance due to the dissemination of antibiotic-resistant strains. Evidence suggests that the ability to form matrix-encased biofilms contributes to the pathogenesis of S. aureus and S. epidermidis. In this study, we investigated the functions of two staphylococcal biofilm matrix polymers: poly-N-acetylglucosamine surface polysaccharide (PNAG) and extracellular DNA (ecDNA). We measured the ability of a PNAG-degrading enzyme (dispersin B) and DNase I to inhibit biofilm formation, detach preformed biofilms, and sensitize biofilms to killing by the cationic detergent cetylpyridinium chloride (CPC) in a 96-well microtiter plate assay. When added to growth medium, both dispersin B and DNase I inhibited biofilm formation by both S. aureus and S. epidermidis. Dispersin B detached preformed S. epidermidis biofilms but not S. aureus biofilms, whereas DNase I detached S. aureus biofilms but not S. epidermidis biofilms. Similarly, dispersin B sensitized S. epidermidis biofilms to CPC killing, whereas DNase I sensitized S. aureus biofilms to CPC killing. We concluded that PNAG and ecDNA play fundamentally different structural roles in S. aureus and S. epidermidis biofilms.

scholarly journals Altered proteome of a Burkholderia pseudomallei mutant defective in short-chain dehydrogenase affects cell adhesion, biofilm formation and heat stress tolerance

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8659 ◽  
Author(s):  
Onrapak Reamtong ◽  
Nitaya Indrawattana ◽  
Amporn Rungruengkitkun ◽  
Tipparat Thiangtrongjit ◽  
Taksaon Duangurai ◽  
...  
Keyword(s):  
Heat Stress ◽  
Biofilm Formation ◽  
Burkholderia Pseudomallei ◽  
Deletion Mutant ◽  
Stress Conditions ◽  
Host Cells ◽  
Short Chain ◽  
Wild Type ◽  
Short Chain Dehydrogenase

Burkholderia pseudomallei is a Gram-negative bacillus that causes melioidosis and is recognized as an important public health problem in southeast Asia and northeast Australia. The treatment of B. pseudomallei infection is hampered by resistance to a wide range of antimicrobial agents and no vaccine is currently available. At present, the underlying mechanisms of B. pseudomallei pathogenesis are poorly understood. In our previous study, we reported that a B. pseudomallei short-chain dehydrogenase (SDO; BPSS2242) mutant constructed by deletion mutagenesis showed reduced B. pseudomallei invasion and initial intracellular survival. This indicated that SDO is associated with the pathogenesis of melioidosis. In the present study, the role of B. pseudomallei SDO was further investigated using the SDO deletion mutant by a proteomic approach. The protein profiles of the SDO mutant and wild-type K96243 were investigated through gel-based proteomic analysis. Quantitative intensity analysis of three individual cultures of the B. pseudomallei SDO mutant revealed significant down-regulation of five protein spots compared with the wild-type. Q-TOF MS/MS identified the protein spots as a glutamate/aspartate ABC transporter, prolyl-tRNA synthetase, Hsp70 family protein, quinone oxidoreductase and a putative carboxypeptidase. Functional assays were performed to investigate the role of these differentially expressed proteins in adhesion to host cells, biofilm induction and survival under heat stress conditions. The SDO deletion mutant showed a decreased ability to adhere to host cells. Moreover, biofilm formation and the survival rate of bacteria under heat stress conditions were also reduced in the mutant strain. Our findings provide insight into the role of SDO in the survival and pathogenesis of B. pseudomallei at the molecular level, which may be applied to the prevention and control of B. pseudomallei infection.

scholarly journals Role of SpoVA Proteins in Release of Dipicolinic Acid during Germination of Bacillus subtilis Spores Triggered by Dodecylamine or Lysozyme

2006 ◽  
Vol 189 (5) ◽  
pp. 1565-1572 ◽  
Author(s):  
Venkata Ramana Vepachedu ◽  
Peter Setlow
Keyword(s):  
Bacillus Subtilis ◽  
Small Molecules ◽  
Spore Germination ◽  
Dipicolinic Acid ◽  
Temperature Sensitive ◽  
Wild Type ◽  
Ph Optimum ◽  
Inner Membrane ◽  
Temperature Sensitive Mutants

ABSTRACT The release of dipicolinic acid (DPA) during the germination of Bacillus subtilis spores by the cationic surfactant dodecylamine exhibited a pH optimum of ∼9 and a temperature optimum of 60°C. DPA release during dodecylamine germination of B. subtilis spores with fourfold-elevated levels of the SpoVA proteins that have been suggested to be involved in the release of DPA during nutrient germination was about fourfold faster than DPA release during dodecylamine germination of wild-type spores and was inhibited by HgCl2. Spores carrying temperature-sensitive mutants in the spoVA operon were also temperature sensitive in DPA release during dodecylamine germination as well as in lysozyme germination of decoated spores. In addition to DPA, dodecylamine triggered the release of amounts of Ca2+ almost equivalent to those of DPA, and at least one other abundant spore small molecule, glutamic acid, was released in parallel with Ca2+ and DPA. These data indicate that (i) dodecylamine triggers spore germination by opening a channel in the inner membrane for Ca2+-DPA and other small molecules, (ii) this channel is composed at least in part of proteins, and (iii) SpoVA proteins are involved in the release of Ca2+-DPA and other small molecules during spore germination, perhaps by being a part of a channel in the spore's inner membrane.

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.

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