scholarly journals Isolation of Shewanella putrefaciens GRD 03 from Fish and Explication of Biofilm Adherence Potency on Different Substrates

2022 ◽  
Author(s):  
Swathy Krishna Jayalekshmi ◽  
Arya Radhakrishnan Krishna ◽  
Trisha Mary Pandipilly Antony ◽  
Suganthi Ramasamy
Keyword(s):  
Foodborne Pathogens ◽  
Biofilm Formation ◽  
Food Poisoning ◽  
Brain Heart Infusion ◽  
Microtiter Plate ◽  
Shewanella Putrefaciens ◽  
Luria Bertani ◽  
Growth Media ◽  
Luria Bertani Broth ◽  
Strong Attachment

Foodborne pathogens are the main threat and cause of food poisoning. The majority of food infections have been related to the biofilm formation of foodborne pathogens in the food industry. Shewanella putrefaciens (KX355803, GRD 03), a Gram-negative pathogen isolated from mackerel fish, was identified and recognized as a food spoilage bacterium and a strong biofilm producer. The adhesion or attachment ability of Shewanella putrefaciens was determined on steel, plastic, glass, PVC and wood. NB (Nutrient broth), LB (Luria-Bertani broth), TSB (Tryptic soy broth) and BHI (Brain heart infusion broth) were enriched with glucose and shows optimum for bacterial adhesion. In the microtiter plate method (MTP), the strong attachment was observed at 48 and 72 hours of incubation and significant differences were obtained at p < 0.05. As the incubation period increases, the OD value (Optical density) of samples also increase. Biofilm formation is the major cause cross-contamination, and shows resistance to certain disinfectants, which leads to environmental stress tolerance. This study suggested with optimum biofilm production of isolate from fish by using glucose enriched media on different substrates, also comparing different growth media provide a detailed idea about biofilm-forming ability at different incubation time intervals.

scholarly journals Pembentukan Biofilm Pseudomonas aeruginosa pada Beberapa Media Cair

2021 ◽  
Vol 10 (2) ◽  
pp. 35-40
Author(s):  
Didik Wahyudi ◽  
Endang Sutariningsih Soetarto
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Optical Density ◽  
Crystal Violet ◽  
Brain Heart Infusion ◽  
Microtiter Plate ◽  
Culture Technique ◽  
Luria Bertani ◽  
Nutrient Broth ◽  
Luria Bertani Broth ◽  
Plate Reader

Pseudomonas aeruginosa merupakan bakteri Gram negatif berbentuk batang bersifat patogen oportunistik yang menjadi penyebab utama infeksi nosokomial dan mampu membentuk biofilm pada media pertumbuhan, biofilm sering mengakibatkan pengobatan penyakit infeksi menjadi lebih sulit.  Media pertumbuhan bakteri ada beberapa jenis, komposisi dan merek. Tujuan penelitian ini adalah untuk mengetahui kemampuan P. aeruginosa dalam membentuk biofilm pada beberapa media biakan cair. P. aeruginosa diisolasi dari sampel klinis dari rumah sakit, media cair yang digunakan adalah nutrien broth, laktosa broth, brain-heart infusion (BHI), luria bertani broth, dan tripticase soy broth.  Uji pembentukan biofilm menggunakan metode microtiter plate culture technique, kemampuan pembetukan biofilm diukur berdasarakan optical density dengan menggunakan microtiter plate reader pada panjang gelombang 570nm, dengan pewarnaan crystal violet 0,1%, setelah inkubasi 24 jam pada suhu 37oC, dengan replikat 8 kali.  Hasil penelitian menunjukkan bahwa P. aeruginosa memiliki kemampuan membentuk biofilm pada nutrient broth 0,926±0,081, laktosa broth 0,521±0,041, BHI 1,283±0,031, luria bertani 1,301±0,043, dan media trypticase soy broth 1,563±0,032.  Pembentukan biofilm tertinggi pada trypticase soy broth, dan terendah pada laktosa broth, sedangkan pada media BHI dan luria bertani kemampuan pembentukan biofilm yang setara.  Kesimpulan penelitian ini adalah P. aeruginosa memiliki kemampuan yang berbeda dalam membentuk biofilm ketika ditumbuhkan pada media cair yang berbeda.Kata kunci : Biofilm, Pseudomonas aeruginosa, media cair

Growth History Influences Starvation-Induced Expression of uspA, grpE, and rpoS and Subsequent Cryotolerance in Escherichia coli O157:H7

2005 ◽  
Vol 68 (6) ◽  
pp. 1154-1158 ◽  
Author(s):  
PURUSHOTTAM V. GAWANDE ◽  
MANSEL W. GRIFFITHS
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogens ◽  
Molecular Mechanisms ◽  
Frozen Storage ◽  
Luria Bertani ◽  
Growth Media ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Luria Bertani Broth ◽  
And Control

In this study, we investigated the effect of starvation on cryotolerance of Escherichia coli O157:H7 grown in tryptic soy broth (TSB) and Luria-Bertani broth (LB). Starved cells (cells suspended in water at 37°C for 6 h) and control cells (cells in TSB or LB) were frozen at −18°C for up to 240 h in their respective growth media. The E. coli grown in TSB showed a greater starvation effect (the difference in percent survival of starved and control cells) and cryotolerance. The starved E. coli grown in TSB showed a 30% increase in their ability to survive frozen storage for 24 h at −18°C. The corresponding increase in survival for LB-grown E. coli was only 3.8%. Cryotolerance induced by starvation of TSB- and LB-grown E. coli was correlated with the expression of genes involved in general stress response pathways, such as uspA, grpE, and rpoS. The expression of uspA, grpE, and rpoS was quantified by measuring the green fluorescence generated from autofluorescent E. coli harboring puspA::gfp, pgrpE::gfp, and prpoS::gfp gene fusions. The results obtained in this study indicate that uspA, grpE, and rpoS were induced on starvation when E. coli was grown in TSB, and their expression correlated well with subsequent induction of cryotolerance developed at −18°C. In contrast, cells grown in LB and subsequently exposed to starvation conditions showed no increase in expression of uspA, grpE, or rpoS, and, as expected, these cells did not exhibit increased cryotolerance at −18°C. Knowledge of molecular mechanisms involved in cross-protection might make it possible to devise strategies to limit their effects and lead to ways to predict the survival of foodborne pathogens in stressful environments.

scholarly journals High-Salt Stress Conditions Increase the pAW63 Transfer Frequency in Bacillus thuringiensis

2012 ◽  
Vol 78 (19) ◽  
pp. 7128-7131 ◽  
Author(s):  
Elise Beuls ◽  
Pauline Modrie ◽  
Cédric Deserranno ◽  
Jacques Mahillon
Keyword(s):  
Salt Stress ◽  
Bacillus Thuringiensis ◽  
Positive Impact ◽  
Brain Heart Infusion ◽  
Luria Bertani ◽  
Plasmid Transfer ◽  
High Salt ◽  
Content Type ◽  
Luria Bertani Broth ◽  
High Salt Stress

ABSTRACTConjugation experiments withBacillus thuringiensisand transfer kinetics demonstrated that salt stress has a positive impact on plasmid transfer efficiency. Compared to standard osmotic conditions (0.5% NaCl), plasmid transfer occurred more rapidly, and at higher frequencies (>100-fold), when bacteria were exposed to a high-salt stress (5% NaCl) in liquid brain heart infusion (BHI). Under milder salt conditions (2.5% NaCl), only a 10-fold effect was observed in Luria-Bertani broth and no difference was detected in BHI. These observations are particularly relevant in the scope of potential gene exchanges among members of theBacillus cereusgroup, which includes food-borne contaminants and pathogens.

scholarly journals Biofilm Formation and Interaction with the Surfaces of Gallstones by Salmonella spp.

2002 ◽  
Vol 70 (5) ◽  
pp. 2640-2649 ◽  
Author(s):  
A. M. Prouty ◽  
W. H. Schwesinger ◽  
J. S. Gunn
Keyword(s):  
Salmonella Enterica ◽  
Biofilm Formation ◽  
Asymptomatic Carrier ◽  
Carrier State ◽  
Luria Bertani ◽  
Salmonella Spp ◽  
Human Gallbladder ◽  
Luria Bertani Broth ◽  
High Concentrations

ABSTRACT Salmonellae can exist in an asymptomatic carrier state in the human gallbladder. Individuals with gallstones are more likely to become typhoid carriers, and antibiotic treatments are often ineffectual against Salmonella enterica serovar Typhi in carriers with gallstones. Therefore, we hypothesized that Salmonella spp. form biofilms on the surfaces of gallstones, where the bacteria are protected from high concentrations of bile and antibiotics. A number of methods were utilized to examine biofilm formation on human gallstones and glass coverslips in vitro, including confocal, light, and scanning electron microscopy. In our assays, salmonellae formed full biofilms on the surfaces of gallstones within 14 days and appeared to excrete an exopolysaccharide layer that bound them to the surfaces and to other bacteria. Efficient biofilm formation on gallstones was dependent upon the presence of bile, as a biofilm did not form on gallstones within 14 days in Luria-Bertani broth alone. The biofilms formed by a Salmonella enterica serovar Typhi Vi antigen mutant, as well as strains with mutations in genes that eliminate production of four different fimbriae, were indistinguishable from the biofilms formed by the parents. Mutants with an incomplete O-antigen, mutants that were nonmotile, and mutants deficient in quorum sensing were unable to develop complete biofilms. In addition, there appeared to be selectivity in salmonella binding to the gallstone surface that did not depend on the topology or surface architecture. These studies should aid in the understanding of the Salmonella carrier state, an important but underresearched area of typhoid fever pathogenesis. If the basis of carrier development can be understood, it may be possible to identify effective strategies to prevent or treat this chronic infection.

scholarly journals Biofilm formation and control strategies of foodborne pathogens: food safety perspectives

RSC Advances ◽  
2017 ◽  
Vol 7 (58) ◽  
pp. 36670-36683 ◽  
Author(s):  
Xihong Zhao ◽  
Fenghuan Zhao ◽  
Jun Wang ◽  
Nanjing Zhong
Keyword(s):  
Food Safety ◽  
Foodborne Pathogens ◽  
Biofilm Formation ◽  
Control Strategies ◽  
Food Poisoning ◽  
Foodborne Diseases ◽  
Main Factors ◽  
And Control

Foodborne pathogens are the main factors behind foodborne diseases and food poisoning and thus pose a great threat to food safety.

scholarly journals Optimum cultural conditions to achieve the best biofilm formation and high level icaA transcription by Staphylococcus aureus

2021 ◽  
Author(s):  
Aram Sharifi ◽  
Abdolmajid Mohammadzadeh ◽  
Pezhman Mahmoodi ◽  
Taghi Zahraei Salehi
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Culture Media ◽  
Brain Heart Infusion ◽  
Microtiter Plate ◽  
Broth Culture ◽  
Microtiter Plates ◽  
Cultural Conditions ◽  
Slime Layer ◽  
High Level

Abstract Background The aim of this study was to investigate the influences of different broth culture media supplemented with glucose, on the biofilm formation and ica expression of Staphylococcus aureus. The phenotypic ability to adhere to a polystyrene surface and to produce slime layer were evaluated using microtiter plate test (MtP) and Congo red tube test, respectively. Using PCR, the presence of ica locus in S. aureus strains was confirmed and subsequently, quantitative real-time RT-PCR was performed to investigate transcription of icaA in various media including Tryptic soy broth (TSB), Brain-heart infusion broth (BHIB), (Nutrient broth) NB and (Muller-Hinton broth) MHB contained 0, 0.25, 0.5, 1 and 2% glucose. Results Our results showed that although all of the studied strains adhered to the wells of polystyrene microtiter plates, the optimum rate of biofilm formation was observed for TSB medium contained 1% glucose, but biofilm formation was not significantly different in NB, MHB and BHIB media. Supplementation of all media with 1% glucose led to the highest production of biofilm formation and in all of media transcription of icaA was increased with glucose addition to one present. Conclusions The results of the present study indicated that TSB medium supplemented with 1% glucose was the most appropriate medium for evaluation of biofilm formation by S. aureus isolates.

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.

Biofilm Formation, Cellulose Production, and Curli Biosynthesis by Salmonella Originating from Produce, Animal, and Clinical Sources†

2005 ◽  
Vol 68 (5) ◽  
pp. 906-912 ◽  
Author(s):  
ETHAN B. SOLOMON ◽  
BRENDAN A. NIEMIRA ◽  
GERALD M. SAPERS ◽  
BASSAM A. ANNOUS
Keyword(s):  
Crystal Violet ◽  
Congo Red ◽  
Salmonella Enterica ◽  
Biofilm Formation ◽  
Binding Assay ◽  
Luria Bertani ◽  
Clinical Isolates ◽  
Cellulose Production ◽  
Luria Bertani Broth ◽  
Static Conditions

The ability of 71 strains of Salmonella enterica originating from produce, meat, or clinical sources to form biofilms was investigated. A crystal violet binding assay demonstrated no significant differences in biofilm formation by isolates from any source when tested in any of the following three media: Luria-Bertani broth supplemented with 2% glucose, tryptic soy broth (TSB), or 1/20th-strength TSB. Incubation was overnight at 30°C under static conditions. Curli production and cellulose production were monitored by assessing morphotypes on Luria-Bertani agar without salt containing Congo red and by assessing fluorescence on Luria-Bertani agar containing calcofluor, respectively. One hundred percent of the clinical isolates exhibited curli biosynthesis, and 73% demonstrated cellulose production. All meat-related isolates formed curli, and 84% produced cellulose. A total of 80% of produce-related isolates produced curli, but only 52% produced cellulose. Crystal violet binding was not statistically different between isolates representing the three morphotypes when grown in TSB; however, significant differences were observed when strains were cultured in the two other media tested. These data demonstrate that the ability to form biofilms is not dependent on the source of the test isolate and suggest a relationship between crystal violet binding and morphotype, with curli- and cellulose-deficient isolates being least effective in biofilm formation.

scholarly journals Glucose and Nutrient Concentrations Affect the Expression of a 104-Kilodalton Listeria Adhesion Protein in Listeria monocytogenes

2002 ◽  
Vol 68 (10) ◽  
pp. 4876-4883 ◽  
Author(s):  
Ziad W. Jaradat ◽  
Arun K. Bhunia
Keyword(s):  
Listeria Monocytogenes ◽  
Brain Heart Infusion ◽  
Luria Bertani ◽  
Nutrient Concentrations ◽  
Enzyme Linked Immunosorbent Assay ◽  
Growth Media ◽  
Adhesion Protein ◽  
Sugar Fermentation ◽  
Rich Media ◽  
High Concentrations

ABSTRACT Growth media and environmental conditions influence the expression of adhesion and invasion proteins in Listeria monocytogenes. Here, the expression of the 104-kDa Listeria adhesion protein (LAP) was studied in nutrient-rich media (Trypticase soy broth [TSB] and brain heart infusion [BHI]), minimal medium (Luria-Bertani [LB]), or nutrient-deficient medium (peptone water [PW]) by immunoblotting, enzyme-linked immunosorbent assay (ELISA), and immunoelectron microscopy. Also, the effect of incorporating different concentrations of glucose on LAP expression was studied. Immunoblotting showed that LAP expression was at least twofold higher in LB medium than in TSB or BHI, while PW supported very poor cell growth and LAP expression. ELISA and immunoblotting results showed that higher concentrations of glucose (>1.6 g/liter) lowered the culture pH and suppressed LAP expression by more than 75%; however, the addition of K2HPO4 reduced this effect. L. monocytogenes cells grown in LB media with lower concentrations of glucose showed higher adhesion to Caco-2 cells (3,716 and 4,186 cpm of attached bacteria for 0 and 0.2 g of glucose/liter, respectively), while L. monocytogenes cells grown in LB with higher glucose concentrations exhibited lower adhesion (2,126 and 2,221 cpm for 1.6 and 3.2 g of glucose/liter, respectively). A LAP-negative L. monocytogenes strain (A572) showed low adhesion profiles regardless of the amount of glucose added. Transmission electron microscopy revealed that LAP is localized mainly in the cytoplasm, with only a few molecules located on the cell surface. Growth in LB with high glucose (3.2 g/liter) showed the presence of only a few molecules in the cells, corroborating the results observed with ELISA or immunoblotting. In summary, nutrient-rich media and high concentrations of glucose suppressed LAP expression, which possibly is due to the changes in the pH of the media during growth from the accumulation of sugar fermentation by-products.

scholarly journals Production of 7-methylxanthine from Theobromine by Metabolically Engineered E. coli

2020 ◽  
Vol 21 (3) ◽  
pp. 19-27
Author(s):  
Khalid Hussein Rheima Algharrawi ◽  
Mani Subramanian
Keyword(s):  
Potassium Phosphate ◽  
Reaction Medium ◽  
Luria Bertani ◽  
Preparative Chromatography ◽  
Growth Media ◽  
Scale Production ◽  
E Coli ◽  
Luria Bertani Broth ◽  
Product Solution ◽  
Biocatalytic Process

In this work, a novel biocatalytic process for the production of 7-methylxanthines from theobromine, an economic feedstock has been developed. Bench scale production of 7-methlxanthine has been demonstrated. The biocatalytic process used in this work operates at 30 OC and atmospheric pressure, and is environmentally friendly. The biocatalyst was E. coli BL21(DE3) engineered with ndmB/D genes combinations. These modifications enabled specific N7- demethylation of theobromine to 7-methylxanthine. This production process consists of uniform fermentation conditions with a specific metabolically engineered strain, uniform induction of specific enzymes for 7-methylxanthine production, uniform recovery and preparation of biocatalyst for reaction and uniform recovery of pure 7-methylxanthine.    Many E. coli BL21(DE3) strains metabolically engineered with single and/or multiple ndmB/D genes were tested for catalytic activity, and the best strains which had the higher activity were chosen to carry out the N-demethylation reaction of theobromine. Strain pBD2dDB had the highest activity for the production of 7-methylxanthine from theobromine. That strain was used to find the optimum amount of cells required to achieve complete conversion of theobromine to 7-methylxanthine within two hours. It was found that the optimum concentration of pBD2dDB strain to achieve 100% conversion of 0.5 mM theobromine to 7-methylxanthine was 5 mg/mL. The cell growth of pBD2dDB strain was studied using two different growth media, (Luria-Bertani Broth and Super Broth). Super broth was found to be the best medium to produce the highest amount of cell paste (1.5 g). Subsequently, the process was scaled up in which 2 L reaction volume was used to produce 7-methylxanthine (100% conversion) from 0.5 mM theobromine catalyzed by pBD2dDB strain. The reactions was carried out at 30 oC and 250 rpm shaker speed, and the reaction medium was 50 mM potassium phosphate buffer (pH=7). 7-methylxanthines was separated by preparative chromatography with high recovery, and the product solution was collected, purified by drying at 120-140 oC for 4 hours and, recovered (127 mg). Purity of the isolated 7-methylxanthine was comparable to authentic standards with no contaminant peaks, as observed by HPLC, LC-MS, and NMR. 

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