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

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 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 Enumeration, Resuscitation, and Infectivity of the Sublethally Injured Erwinia Cells Induced by Mild Acid Treatment

2004 ◽  
Vol 94 (1) ◽  
pp. 76-81 ◽  
Author(s):  
Ching-Hsing Liao ◽  
Lisa M. Shollenberger
Keyword(s):  
Crystal Violet ◽  
Disease Ecology ◽  
Sublethal Effects ◽  
Current Knowledge ◽  
Sodium Dodecyl ◽  
Brain Heart Infusion ◽  
Soft Rot ◽  
Nutrient Broth ◽  
Selective Media ◽  
Cucumber Fruit

Current knowledge about microbial injury was derived mostly from studies with bacteria associated with processed foods. Demonstration of injury and repair in phytopathogenic bacteria and its implication on pathogen detection and disease ecology have not been reported. The objective of this study was to investigate the lethal and sublethal effects of acetic acid (AA) on soft-rot Erwinia spp. and the limitation of using selective media for isolation of injured cells. Following exposure to 0.3% AA for 6 min, 90 to 99% of E. carotovora subsp. carotovora, E. carotovora subsp. atroseptica, and E. chrysanthemi cells were killed. When AA-treated samples were plated on agar media, the number of bacteria recovered on nonselective media such as brain heart infusion agar was 3 log units higher than that recovered on selective media such as crystal violet pectate (CVP). Lethal and sublethal actions of AA on Erwinia spp. were affected greatly by acid concentration, exposure time, and bacterial strains tested. Injured Erwinia cells were able to repair and regain the ability to grow on CVP when placed in nutrient broth but not in selective broth containing crystal violet and sodium dodecyl sulfate. Injured cells also were able to resuscitate on cut surfaces of cucumber fruit and to induce soft rot on green bell pepper. Together, these results suggest that direct use of selective media for isolation of Erwinia spp. could limit the recovery of injured cells in samples that have been exposed to chemical or physical stresses. Enrichment of these samples in nutrient broth before plating on CVP is expected to improve the detection of injured Erwinia spp.

Efektivitas Antibakteri Ekstrak Daun Mangrove Acanthus ilicifolius Terhadap Biofilm Enterococcus faecalis

DENTA ◽  
2015 ◽  
Vol 9 (2) ◽  
pp. 136
Author(s):  
Hariningtyas Dian Rachmawati ◽  
A Aprilia ◽  
Kristanti Parisihni
Keyword(s):  
Enterococcus Faecalis ◽  
Optical Density ◽  
Crystal Violet ◽  
Microtiter Plate ◽  
Control Group ◽  
Control Group Design ◽  
Group Design ◽  
Acanthus Ilicifolius ◽  
Post Test ◽  
Post Hoc

<strong>Latar Belakang: </strong><em>Enterococcus faecalis</em> adalah salah satu bakteri yang memiliki kemampuan dalam membentuk biofilm yang dapat menyebabkan infeksi persisten endodontik. Biofilm adalah kumpulan bakteri yang terorganisasi dengan baik yang melekat pada permukaan dan terlapisi oleh lapisan matriks ekstraselular polisakarida. Ekstrak daun mangrove <em>Acanthus ilicifolius </em>telah diketahui memiliki senyawa bioaktif yang berpotensi sebagai antibakteri seperti saponin, alkaloid, terpenoid, dan tanin. <strong>Tujuan:</strong> Untuk mengetahui efektivitas antibakteri ekstrak daun mangrove <em>Acanthus ilicifolius</em> terhadap biofilm <em>Enterococcus faecalis.</em> <strong>Metode</strong> : Penelitian ini merupakan penelitian eksperimental dengan desain penelitian <em>post test only control group design</em>. Sampel penelitian ini menggunakan bakteri <em>Enterococcus faecalis, </em>dibagi menjadi 6 kelompok, terdiri dari kontrol positif (NaOCL 2,5%), dan 5 kelompok perlakuan dengan konsentrasi ekstrak daun mangrove <em>Acanthus ilicifoliu</em>s yang berbeda yaitu 60 mg/ml; 70 mg/ml; 80 mg/ml; 90 mg/ml; dan 100 mg/ml dan dilakukan pengulangan sebanyak 8 kali. Bakteri <em>Enterococcus faecalis </em>dikultur pada media <em>TSBglu</em> diinkubasi selama semalam<em>.</em> Sebanyak 0,1 ml bakteri <em>Enterococcus faecalis</em> dengan konsentrasi 10<sup>6</sup> diisikan pada <em>microtiter plate</em> kemudian<em> </em>di cat dengan <em>crystal violet</em>.<em> </em>Biofilm diperiksa dengan mengukur <em>Optical Density</em> menggunakan ELISA <em>reader</em>. Analisis data menggunakan uji <em>One-way </em>ANOVA dilanjutkan dengan uji <em>Post Hoc</em>. <strong>Hasil:</strong> Pemberian ekstrak daun mangrove <em>Acanthus ilicifolius</em> menurunkan OD biofilm <em>Enterococcus faecalis</em> (p&lt;0,05) pada semua kelompok. <strong>Simpulan: </strong>Ekstrak daun mangrove <em>Acanthus ilicifolius </em>memiliki efektivitas antibakteri terhadap biofilm <em>Enterococcus faecalis</em>

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 Pengaruh Ekstrak Alpinia galanga L Terhadap Produksi Biofilm pada Escherichia coli

2021 ◽  
Vol 10 (1) ◽  
pp. 24-30
Author(s):  
Didik Wahyudi ◽  
Syahran Wael
Keyword(s):  
Escherichia Coli ◽  
Optical Density ◽  
Crystal Violet ◽  
Microtiter Plate ◽  
Alpinia Galanga ◽  
One Way Anova

Escherichia coli merupakan bakteri Gram negatif berbentuk batang, mampu menyebabkan infeksi di beberapa bagian tubuh, dan ditemukan telah resisten terhadap berbagai antibiotic, salah satu factor penyebabnya adalah kemampuannya membentuk biofilm pada jaringan. Alpinia gallanga L memiliki kemampuan menghambat pembentukan biofilm terhadap beberapa bakteri. Tujuan Penelitian ini adalah untuk mengetahui kemampuan ekstrak Alpinia galangal L dalam menghambat produksi biofilm Escherichia coli. Penelitian ini di awali dengan ekstraksi rimpang lengkuas dengan etanol menggunakan metode masersi, kemudian dibuat konsentrasi 10%, 20%, 30%, 40%, dan 50%. Escherichia coli diisolasi dari kasus Infeksi Saluran Kemih (ISK). kemudian di lakukan karakterisasi fisiologisnya dan uji kepekaan terhadap antibiotik. Uji penghambatan biofilm Escherichia coli dilakukan dengan metode microtiter plate culture dengan menggunakan spektrofotometer pada panjang gelombang 595nm, Hasil pengukuran produksi biofilm berupa besarnya nilai Optical Density crystal violet 0,1%, setiap perlakuan menggunakan ulangan 8 kali, data yang didapatkan dianalisis dengan One Way Anova. Hasil Penelitian menunjukan bahwa Ekstrak Alpinia galanga L mampu menghambat produksi biofilm Escherichia coli pada konsentrasi 30%.

scholarly journals Deteksi Acinetobacter baumannii Multiresisten Obat Penghasil Biofilm menggunakan Pewarnaan Berbasis Crystal Violet

2019 ◽  
Vol 5 (2) ◽  
pp. 41
Author(s):  
Ardiana Kusumaningrum ◽  
Iin Maemunah ◽  
Dimas Seto Prasetyo ◽  
Fera Ibrahim ◽  
Budiman Bela
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Acinetobacter Baumannii ◽  
Crystal Violet ◽  
Luria Bertani

Pendahuluan: Kasus infeksi terkait biofilm merupakan masalah besar pada dunia kesehatan karena sifat kekebalannya terhadap antibiotik dan respon imun, terutama pada kasus infeksi kronik akibat Acinetobacter baumannii. Saat ini terdapat berbagai metode deteksi pembentukan biofilm, namun pemeriksaan tersebut belum dilakukan secara rutin karena berbagai keterbatasan. Oleh karena itu, perlu dilakukan optimasi metode deteksi biofilm menggunakan bahan yang rutin tersedia di laboratorium serta mengukur proporsi sampel yang mengandung bakteri penghasil biofilm Metode: Desain penelitian ini adalah uji eksperimental. Pada penelitian ini dilakukan pengembangan deteksi pembentukan biofilm menggunakan metoda tabung microcentrifuse tube 1,5 ml berbahan polypropylene dengan zat warna berbasis crystal violet konsentrasi 0,1% dan 1%. Optimasi yang dilakukan meliputi media biakan, lama inkubasi, inokulum bakteri yang digunakan serta bahan pembilas. Isolat bakteri Pseudomonas aeruginosa ATCC 10145, A. baumannii ATCC 19606  dan Staphylococcus aureus ATCC 25923 digunakan sebagai kontrol positif dan kontrol negatif pemeriksaan yang dilakukan. Hasil: Pemeriksaan optimasi yang dilakukan, diperoleh kondisi optimal pembentukan biofilm menggunakan media Luria Bertani dengan besar inokulum 1 sengkelit penuh, lama inkubasi 30 jam dan pewarnaan dilakukan menggunakan crystal violet 0,1% serta bahan pembilas berupa PBS steril. Proporsi pembentukan biofilm pada A. baumannii multiresisten obat sebesar 55,3%. Kesimpulan: Metode deteksi pembentukan biofilm menggunakan metode tabung polypropylene yang dimodifikasi dan pewarnaan zat warna crystal violet 0,1% merupakan metode deteksi yang mudah dikerjakan, reproducible dan efisien, sehingga dapat dilakukan di laboratorium mikrobiologi klinik sederhana. Proporsi bakteri penghasil biofilm adalah lebih dari 50% A. baumannii resisten multiobat.

scholarly journals Insight into the Metabolic Profiles of Pb(II) Removing Microorganisms

Molecules ◽  
2021 ◽  
Vol 26 (13) ◽  
pp. 4008
Author(s):  
Carla Cilliers ◽  
Evans M. N. Chirwa ◽  
Hendrik G. Brink
Keyword(s):  
Liquid Chromatography ◽  
High Performance ◽  
Microbial Consortium ◽  
Luria Bertani ◽  
Nutrient Concentrations ◽  
Batch Experiments ◽  
Metabolic Study ◽  
Luria Bertani Broth ◽  
Tandem Mass Spectroscopy ◽  
Insight Into

The objective of the study was to gather insight into the metabolism of lead-removing microorganisms, coupled with Pb(II) removal, biomass viability and nitrate concentrations for Pb(II) bioremoval using an industrially obtained microbial consortium. The consortium used for study has proven to be highly effective at removing aqueous Pb(II) from solution. Anaerobic batch experiments were conducted with Luria-Bertani broth as rich growth medium over a period of 33 h, comparing a lower concentration of Pb(II) with a higher concentration at two different nutrient concentrations. Metabolite profiling and quantification were conducted with the aid of both liquid chromatography coupled with tandem mass spectroscopy (UPLC-HDMS) in a “non-targeted” fashion and high-performance liquid chromatography (HPLC) in a “targeted” fashion. Four main compounds were identified, and a metabolic study was conducted on each to establish their possible significance for Pb(II) bioremoval. The study investigates the first metabolic profile to date for Pb(II) bioremoval, which in turn can result in a clarified understanding for development on an industrial and microbial level.

scholarly journals Arginine Inhibits Serpins

2007 ◽  
Vol 13 (2) ◽  
pp. 213-218
Author(s):  
Thomas W. Stief
Keyword(s):  
Esterase Activity ◽  
Serine Proteases ◽  
Biological Fluid ◽  
Microtiter Plate ◽  
Human Albumin ◽  
Amidolytic Activity ◽  
Peptide Substrates ◽  
Plate Reader ◽  
High Concentrations ◽  
Very High

Serine protease inactivators (serpins) are important regulators in biochemistry. Often it is necessary to block the serpin action, that is, to stabilize the sample. The guanidine group of arginine is the ligand for the active center pocket of many serine proteases. Arginine or guanidine inhibits serine proteases, and arginine belongs to the reactive P1-P1' center of many serpins. The plasmatic antithrombin, antiplasmin, or anti-C1-esterase activity was determined: A total of 20 µL of pooled normal plasma or 7% human albumin was added to 100 µL of 0—2.67 M arginine, pH 8.6, 10 µL of 26 mIU/mL thrombin in 7% human albumin, and 30 µL of 1.7 mM CHG-Ala-Arg-pNA (37°C). ΔA at 405 nm was determined, by using a microtiter plate reader. Thrombin was substituted by plasmin or C1-esterase, and the chromogenic peptide substrates <Glu-Phe-Lys-pNA or MeOC-Lys(eCBO)-Gly-Arg-pNA, respectively, were used. The IC50 of arginine against plasmatic antithrombin activity is 580 mM; the IC 25 is 440 mM. The IC25 of arginine against plasmatic α 2-antiplasmin or C1-inactivator is 1650 mM. The amidolytic activity of thrombin, plasmin, and C1-esterase is inhibited similarly by arginine: the IC50 for arginine against the amidolytic activity of these proteases is about 400 mM. Arginine at very high concentrations inhibits serpins. This is important, if stabilization of a biological fluid is a prerequisite for valid activities of serine proteases. In addition, these high concentrations of arginine might be a new gentle principle to inhibit pathogens that need serpins for their pathophysiology.

scholarly journals Bicarbonate Evokes Reciprocal Changes in Intracellular Cyclic di-GMP and Cyclic AMP Levels in Pseudomonas aeruginosa

Biology ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 519
Author(s):  
Kasidid Ruksakiet ◽  
Balázs Stercz ◽  
Gergő Tóth ◽  
Pongsiri Jaikumpun ◽  
Ilona Gróf ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cyclic Amp ◽  
Biofilm Formation ◽  
Second Messengers ◽  
Brain Heart Infusion ◽  
Ambient Air ◽  
Dependent Manner ◽  
Environmental Signals ◽  
Common Causes ◽  
Camp Levels

The formation of Pseudomonas aeruginosa biofilms in cystic fibrosis (CF) is one of the most common causes of morbidity and mortality in CF patients. Cyclic di-GMP and cyclic AMP are second messengers regulating the bacterial lifestyle transition in response to environmental signals. We aimed to investigate the effects of extracellular pH and bicarbonate on intracellular c-di-GMP and cAMP levels, and on biofilm formation. P. aeruginosa was inoculated in a brain–heart infusion medium supplemented with 25 and 50 mM NaCl in ambient air (pH adjusted to 7.4 and 7.7 respectively), or with 25 and 50 mM NaHCO3 in 5% CO2 (pH 7.4 and 7.7). After 16 h incubation, c-di-GMP and cAMP were extracted and their concentrations determined. Biofilm formation was investigated using an xCelligence real-time cell analyzer and by crystal violet assay. Our results show that HCO3− exposure decreased c-di-GMP and increased cAMP levels in a dose-dependent manner. Biofilm formation was also reduced after 48 h exposure to HCO3−. The reciprocal changes in second messenger concentrations were not influenced by changes in medium pH or osmolality. These findings indicate that HCO3− per se modulates the levels of c-di-GMP and cAMP, thereby inhibiting biofilm formation and promoting the planktonic lifestyle of the bacteria.

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