Pengaruh Ekstrak Alpinia galanga L Terhadap Produksi Biofilm pada Escherichia coli

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%.

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Potensi kitosan kepiting rajungan (Portunus pelagicus) dalam penghambatan pembentukan biofilm Porphyromonas gingivalis dan pertumbuhan Candida albicansPotential of flower crab (Portunus pelagicus) chitosan in the inhibition of Porphyromonas gingivalis and Candida albicans biofilm

Padjadjaran Journal of Dental Researchers and Students ◽

10.24198/pjdrs.v4i2.26636 ◽

2020 ◽

Vol 4 (2) ◽

pp. 121

Author(s):

Yoifah Rizka Wedarti ◽

Laurencia Isabella Loekito ◽

Fani Pangabdian ◽

Dwi Andriani

Keyword(s):

Candida Albicans ◽

Porphyromonas Gingivalis ◽

Optical Density ◽

Biofilm Formation ◽

Microtiter Plate ◽

Control Group ◽

Portunus Pelagicus ◽

Biofilm Inhibition ◽

Significant Difference ◽

One Way Anova

Pendahuluan: Pembentukan biofilm sangat penting dalam patogenesis periodontitis. Porphyromonas gingivalis merupakan bakteri yang banyak ditemukan pada plak gigi dan memiliki kemampuan membentuk biofilm demikian juga Candida albicans memiliki faktor virulensi yang dapat membantu kolonisasi dan proliferasi bakteri di dalam poket periodontal. Ekstrak kitosan kepiting rajungan (Portunus pelagicus) mempunyai potensi antimikrobial yang dapat digunakan sebagai alternatif terapi. Tujuan penelitian ini adalah untuk menganalisis potensi kitosan kepiting rajungan (Portunus pelagicus) dalam penghambatan biofilm Porphyromonas gingivalis dan Candida albicans. Metode: Jenis penelitian adalah eksperimental murni. Penelitian ini menggunakan ekstrak kitosan kepiting rajungan (Portunus pelagicus) terhadap biofilm Porphyromonas gingivalis dan biofilm Candida albicans. Dibagi menjadi 4 kelompok, di mana tiap kelompok terdiri dari 4 sampel. Kelompok K+ (kelompok kontrol positif), P1(kitosan 0,25%), P2 (kitosan 0,5%), P3 (kitosan 1%). Penghambatan biofilm ditentukan dengan menggunakan metode microtiter plate yang menghasilkan nilai optical density kemudian dihitung dengan menggunakan rumus persen penghambatan. Analisis data menggunakan one-way ANOVA diikuti dengan uji LSD. Hasil: Terdapat perbedaan yang signifikan penghambatan biofilm dari kitosan Portunus pelagicus terhadap Porphyromonas gingivalis (p<0,05) antara kelompok, kecuali K + dengan P3. Sedangkan untuk penghambatan Candida albicans menunjukkan bahwa ada perbedaan yang signifikan dalam persentase penghambatan biofilm (p<0,05), antara kelompok K+ dengan P2 dan P3; kelompok P1 dengan P2 dan P3; kelompok P2 dengan P3. Simpulan: Kitosan Portunus pelagicus memiliki potensi dalam menghambat pembentukan biofilm Porphyromonas gingivalis dan pertumbuhan Candida albicans. Kitosan Portunus pelagicus 1% memiliki efek antimikrobial terbesar pada biofilm.Kata kunci: Biofilm, Porphyromonas gingivalis, Candida albicans, kitosan portunus pelagicus, periodontitis. ABSTRACTIntroduction: Biofilm formation is important in periodontitis pathogenesis. Porphyromonas gingivalis and Candida albicans, which are found in dental plaque and can form a biofilm, have virulence factor that facilitates the bacterial colonisation and proliferation in periodontal pockets. Chitosan extract of flower crab (Portunus pelagicus) has antimicrobial potential which can be used as an alternative therapy. The objective of this research was to analyse the potential of flower crab (Portunus pelagicus) chitosan in the inhibition of Porphyromonas gingivalis and Candida albicans biofilms. Methods: This research was a pure experimental laboratory. This study used flower crab (Portunus pelagicus) chitosan to inhibit the biofilm formation of Porphyromonas gingivalis and Candida albicans. The subjects were divided into four groups, where each group consisted of 4 samples. The K+ (positive control group), P1 (0.25% chitosan), P2 (0.5% chitosan), and P3 (1% chitosan). The biofilm inhibition was determined using the microtiter plate methods, which results in the value of optical density, then calculated using the inhibition formula percentage. Data analysis was conducted using the one-way ANOVA followed by the LSD test. Results: There were significant differences in the Porphyromonas gingivalis biofilm inhibition between groups (p < 0.05), except in group K+ with P3. Whereas for Candida albicans biofilm inhibition showed no significant difference (p < 0.05) between group K+ with P2 and P3; group P1 with P2 and P3; and group P2 with P3. Conclusion: The chitosan of flower crab (Portunus pelagicus) has the potential in inhibiting the biofilm formation of Porphyromonas gingivalis and Candida albicans. The highest antibacterial effect on the biofilm formation is shown in the concentration of 1%.Keywords: Biofilm, Porphyromonas gingivalis, Candida albicans, chitosan, Portunus pelagicus, periodontitis.

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Pembentukan Biofilm Pseudomonas aeruginosa pada Beberapa Media Cair

Jurnal Farmasi (Journal of Pharmacy) ◽

10.37013/jf.v10i2.142 ◽

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

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Efektivitas Antibakteri Ekstrak Daun Mangrove Acanthus ilicifolius Terhadap Biofilm Enterococcus faecalis

DENTA ◽

10.30649/denta.v9i2.11 ◽

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

Latar Belakang: Enterococcus faecalis 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 Acanthus ilicifolius telah diketahui memiliki senyawa bioaktif yang berpotensi sebagai antibakteri seperti saponin, alkaloid, terpenoid, dan tanin. Tujuan: Untuk mengetahui efektivitas antibakteri ekstrak daun mangrove Acanthus ilicifolius terhadap biofilm Enterococcus faecalis. Metode : Penelitian ini merupakan penelitian eksperimental dengan desain penelitian post test only control group design. Sampel penelitian ini menggunakan bakteri Enterococcus faecalis, dibagi menjadi 6 kelompok, terdiri dari kontrol positif (NaOCL 2,5%), dan 5 kelompok perlakuan dengan konsentrasi ekstrak daun mangrove Acanthus ilicifolius 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 Enterococcus faecalis dikultur pada media TSBglu diinkubasi selama semalam. Sebanyak 0,1 ml bakteri Enterococcus faecalis dengan konsentrasi 106 diisikan pada microtiter plate kemudian di cat dengan crystal violet. Biofilm diperiksa dengan mengukur Optical Density menggunakan ELISA reader. Analisis data menggunakan uji One-way ANOVA dilanjutkan dengan uji Post Hoc. Hasil: Pemberian ekstrak daun mangrove Acanthus ilicifolius menurunkan OD biofilm Enterococcus faecalis (p<0,05) pada semua kelompok. Simpulan: Ekstrak daun mangrove Acanthus ilicifolius memiliki efektivitas antibakteri terhadap biofilm Enterococcus faecalis

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Enhancement of the Efficiency of Secretion of Heterologous Lipase in Escherichia coli by Directed Evolution of the ABC Transporter System

Applied and Environmental Microbiology ◽

10.1128/aem.71.7.3468-3474.2005 ◽

2005 ◽

Vol 71 (7) ◽

pp. 3468-3474 ◽

Cited By ~ 19

Author(s):

Gyeong Tae Eom ◽

Jae Kwang Song ◽

Jung Hoon Ahn ◽

Yeon Soo Seo ◽

Joon Shick Rhee

Keyword(s):

Escherichia Coli ◽

Directed Evolution ◽

Abc Transporter ◽

Microtiter Plate ◽

Single Amino Acid ◽

Wild Type ◽

E Coli ◽

Single Amino Acid Change ◽

Secretion Efficiency

ABSTRACT The ABC transporter (TliDEF) from Pseudomonas fluorescens SIK W1, which mediated the secretion of a thermostable lipase (TliA) into the extracellular space in Escherichia coli, was engineered using directed evolution (error-prone PCR) to improve its secretion efficiency. TliD mutants with increased secretion efficiency were identified by coexpressing the mutated tliD library with the wild-type tliA lipase in E. coli and by screening the library with a tributyrin-emulsified indicator plate assay and a microtiter plate-based assay. Four selected mutants from one round of error-prone PCR mutagenesis, T6, T8, T24, and T35, showed 3.2-, 2.6-, 2.9-, and 3.0-fold increases in the level of secretion of TliA lipase, respectively, but had almost the same level of expression of TliD in the membrane as the strain with the wild-type TliDEF transporter. These results indicated that the improved secretion of TliA lipase was mediated by the transporter mutations. Each mutant had a single amino acid change in the predicted cytoplasmic regions in the membrane domain of TliD, implying that the corresponding region of TliD was important for the improved and successful secretion of the target protein. We therefore concluded that the efficiency of secretion of a heterologous protein in E. coli can be enhanced by in vitro engineering of the ABC transporter.

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Scale-up from microtiter plate to laboratory fermenter: evaluation by online monitoring techniques of growth and protein expression in Escherichia coli and Hansenula polymorpha fermentations

Microbial Cell Factories ◽

10.1186/1475-2859-8-68 ◽

2009 ◽

Vol 8 (1) ◽

pp. 68 ◽

Cited By ~ 63

Author(s):

Frank Kensy ◽

Christoph Engelbrecht ◽

Jochen Büchs

Keyword(s):

Escherichia Coli ◽

Protein Expression ◽

Online Monitoring ◽

Hansenula Polymorpha ◽

Scale Up ◽

Microtiter Plate ◽

Monitoring Techniques ◽

Expression In Escherichia Coli

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106 ASSESSMENT OF ANTI-BACTERIAL EFFECTS OF PEGYLATED SILVER-COATED CARBON NANOTUBES ON CAUSATIVE BACTERIA OF BOVINE INFERTILITY USING BIOLUMINESCENCE IMAGING SYSTEM

Reproduction Fertility and Development ◽

10.1071/rdv29n1ab106 ◽

2017 ◽

Vol 29 (1) ◽

pp. 161 ◽

Cited By ~ 1

Author(s):

S. Park ◽

A. A. Chaudhari ◽

S. Pillai ◽

S. R. Singh ◽

S. T. Willard ◽

...

Keyword(s):

Escherichia Coli ◽

Carbon Nanotubes ◽

Optical Density ◽

Bacterial Infections ◽

Pathogenic Bacteria ◽

Bioluminescence Imaging ◽

Imaging System ◽

Control Group ◽

Silver Nanotubes ◽

Dose Dependent

Pathogenic bacteria including Escherichia coli and Salmonella sp. are the major causative agents of endometritis and can cause infertility in livestock animals. Antibiotics are commonly used to terminate bacterial infections, but the development of bacterial antibiotic resistance is often encountered. Nanotechnology associated with silver nanoparticles has been highlighted as an alternative anti-bacterial agent, and pegylated silver-coated single-walled carbon nanotubes have high anti-bacterial effects and are non-toxic to human and murine cells in vitro. Here we verified whether a real-time bioluminescence monitoring system could be an alternative tool to assess anti-bacterial effects of nanotubes in a noninvasive approach. Escherichia coli and Salmonella sp. were transfected with plasmids containing constructs for luciferase enzyme (LuxCDABE) and substrate (luciferin) to create self-illuminating bioluminescent bacteria. Pathogens were grown in LB broth at 37°C, adjusted to 107 cfu mL−1, and placed in 96-well plates for treatments. Pegylated (pSWCNTs-Ag) and non-pegylated (SWCNTs-Ag) nanotubes were prepared and added to culture wells at various concentrations (31.25–125 µg mL−1). The control group corresponded to bacteria without nanotubes (0 µg mL−1). Anti-bacterial effects of nanotubes were determined every 10 min until 1 h, then every 30 min up to 6 h incubation through optical density (600 nm) measurements and bioluminescence imaging (BLI) and quantification using an IVIS system. Optical density and BLI data were compared at each time-point using 2-way ANOVA, with P < 0.05 set for significance. Bioluminescence signals emitted by both bacteria stains appeared within 10 min of incubation. Thereafter, control bacteria showed exponential growth that was detected as early as 25 min post-incubation. Bioluminescence imaging revealed dose-dependent anti-bacterial activities of both pSWCNTs-Ag and SWCNTs-Ag on each E. coli and Salmonella sp. (P < 0.05). Contrary to BLI, the OD values did not always reflect bacteria concentrations, and varied according to nanotube concentrations. No significant differences in anti-bacterial activities were revealed between pSWCNTs-Ag and SWCNTs-Ag based on OD values during 6 h of incubation (P > 0.05); meanwhile, pSWCNTs-Ag nanotubes exhibited stronger anti-bacterial effects than SWCNTs-Ag during the same period using BLI (P < 0.05). In summary, we confirmed previous reports showing dose-dependent eliminations of pathogenic bacteria by silver nanotubes. Pegylated nanotubes exhibited high anti-bacterial activity compared to non-pegylated nanotubes. Bioluminescence imaging system revealed superior resolution to enable precise investigation of anti-bacterial kinetics of silver nanotubes. This feature could be useful for the study of bacterial infections that impair livestock fertility. Work was supported by USDA-ARS Biophotonics Initiative grant #58-6402-3-018.

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Expression of Pseudomonas aeruginosa Multidrug Efflux Pumps MexA-MexB-OprM and MexC-MexD-OprJ in a Multidrug-Sensitive Escherichia coli Strain

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.42.1.65 ◽

1998 ◽

Vol 42 (1) ◽

pp. 65-71 ◽

Cited By ~ 119

Author(s):

Ramakrishnan Srikumar ◽

Tatiana Kon ◽

Naomasa Gotoh ◽

Keith Poole

Keyword(s):

Escherichia Coli ◽

Pseudomonas Aeruginosa ◽

Antibiotic Resistance ◽

Crystal Violet ◽

Efflux Pump ◽

Efflux Pumps ◽

Multidrug Efflux ◽

Efflux System ◽

E Coli ◽

Multidrug Efflux Pumps

ABSTRACT The mexCD-oprJ and mexAB-oprM operons encode components of two distinct multidrug efflux pumps inPseudomonas aeruginosa. To assess the contribution of individual components to antibiotic resistance and substrate specificity, these operons and their component genes were cloned and expressed in Escherichia coli. Western immunoblotting confirmed expression of the P. aeruginosa efflux pump components in E. coli strains expressing and deficient in the endogenous multidrug efflux system (AcrAB), although only the ΔacrAB strain, KZM120, demonstrated increased resistance to antibiotics in the presence of the P. aeruginosa efflux genes. E. coli KZM120 expressing MexAB-OprM showed increased resistance to quinolones, chloramphenicol, erythromycin, azithromycin, sodium dodecyl sulfate (SDS), crystal violet, novobiocin, and, significantly, several β-lactams, which is reminiscent of the operation of this pump in P. aeruginosa. This confirmed previous suggestions that MexAB-OprM provides a direct contribution to β-lactam resistance via the efflux of this group of antibiotics. An increase in antibiotic resistance, however, was not observed when MexAB or OprM alone was expressed in KZM120. Thus, despite the fact that β-lactams act within the periplasm, OprM alone is insufficient to provide resistance to these agents. E. coli KZM120 expressing MexCD-OprJ also showed increased resistance to quinolones, chloramphenicol, macrolides, SDS, and crystal violet, though not to most β-lactams or novobiocin, again somewhat reminiscent of the antibiotic resistance profile of MexCD-OprJ-expressing strains ofP. aeruginosa. Surprisingly, E. coli KZM120 expressing MexCD alone also showed an increase in resistance to these agents, while an OprJ-expressing KZM120 failed to demonstrate any increase in antibiotic resistance. MexCD-mediated resistance, however, was absent in a tolC mutant of KZM120, indicating that MexCD functions in KZM120 in conjunction with TolC, the previously identified outer membrane component of the AcrAB-TolC efflux system. These data confirm that a tripartite efflux pump is necessary for the efflux of all substrate antibiotics and that the P. aeruginosa multidrug efflux pumps are functional and retain their substrate specificity in E. coli.

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Transformation of Sinorhizobium meliloti MTCC 100 and Mesorhizobium ciceri TAL 620 by CaCl2 method

Canadian Journal of Microbiology ◽

10.1139/w09-114 ◽

2010 ◽

Vol 56 (1) ◽

pp. 74-76 ◽

Cited By ~ 1

Author(s):

Urmi Patel ◽

Sarika Sinha

Keyword(s):

Escherichia Coli ◽

Optical Density ◽

Sinorhizobium Meliloti ◽

Transformation Efficiency ◽

Binary Vector ◽

Transformation Method ◽

Mesorhizobium Ciceri ◽

Competent Cells

The CaCl2 method, commonly used for transformation of Escherichia coli, was modified and used to develop a simpler and easier transformation method for Rhizobia sp. Two species of Rhizobia, Sinorhizobium meliloti MTCC 100 and Mesorhizobium ciceri TAL 620, were transformed with the 13.2 kb binary vector pGA482. At an optical density of 0.4, the transformation efficiencies in Sinorhizobium meliloti MTCC 100 and Mesorhizobium ciceri TAL 620 were 104 and 103, respectively. Competent cells of Sinorhizobium meliloti MTCC 100 were prepared at different growth intervals and transformed by the same vector. A maximum transformation efficiency of 104 was achieved at an optical density of 0.5.

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