Microtitre Plate Assay for Biofilm Formation, Production and Utilization of Hydroxybiphenyl by Rhodococcus sp. Isolated from Gasoline-Contaminated Soil

2008 ◽  
Vol 63 (7-8) ◽  
pp. 599-604 ◽  
Author(s):  
Zahra Etemadifar ◽  
Giti Emtiazi
Keyword(s):  
Contaminated Soil ◽  
Biofilm Formation ◽  
Carbon Sources ◽  
Rhodococcus Erythropolis ◽  
Rrna Gene ◽  
Sulfur Source ◽  
Microtitre Plate ◽  
Biochemical Tests ◽  
Microtitre Plate Assay ◽  
Microtiter Assay

Gasoline-contaminated soil from Isfahan, Iran was selected to isolate a bacterium capable of desulfurizing dibenzothiophene (DBT). The isolated strain was named R1 and identified as Rhodococcus erythropolis through biochemical tests as well as sequencing of 16S rRNA gene. This strain could efficiently produce 2-hydroxybiphenyl (HBP) from DBT via the 4S metabolic pathway. The highest HBP amount was produced at 2 mm DBT with addition of glucose (10 g l-1), ethanol (3 g l-1), glycerol (2 g l-1) or succinate (10 g l-1) as carbon sources at pH 7. Highest respiration and growth rates were observed by microplate titration on 0.1 mm HBP, and addition of 0.2 mm HBP to glucose (1 g l-1) and DBT (0.3 mm) could inhibite the respiration of the isolate. The isolated strain could grow up to 0.4 mm of HBP when it is used with mineral sulfur as sole sulfur source. To the best of our knowledge this is the first report on a microtiter assay for the production and utilization of HBP by Rhodococcus.

scholarly journals Targeting cellular metabolism to inhibit synergistic biofilm formation of multi-species isolated from a cooling water system

2021 ◽  
Author(s):  
Dingrong Kang ◽  
Wenzheng Liu ◽  
Fatemeh Bajoul kakahi ◽  
Frank Delvigne
Keyword(s):  
Biofilm Formation ◽  
Volatile Fatty Acids ◽  
Species Coexistence ◽  
Synthetic Medium ◽  
Carbon Sources ◽  
Cellular Metabolism ◽  
Metabolic Inhibitors ◽  
Rrna Gene ◽  
Natural Environments ◽  
Bacterial Strains

AbstractBiofilm is ubiquitous in natural environments, causing biofouling in industrial water systems and leading to liquidity and heat transfer efficiency decreases. In particular, multi-species coexistence in biofilms can provide the synergy needed to boost biomass production and enhance treatment resistance. In this study, a total of 37 bacterial strains were isolated from a cooling tower where acetic acid and propionic acid were used as the primary carbon sources. These isolates mainly belonged to Proteobacteria and Firmicutes, which occupied more than 80% of the total strains according to the 16S rRNA gene amplicon sequencing. Four species (Acinetobacter sp. CTS3, Corynebacterium sp. CTS5, Providencia sp. CTS12, and Pseudomonas sp. CTS17) were observed to co-exist in the synthetic medium, showing a synergistic effect towards biofilm formation. Three metabolic inhibitors (sulfathiazole, 3-Bromopyruvic acid, and 3-Nitropropionic acid) were employed as possible treatments against biofilm formation due to their inhibition effect on c-di-GMP biosynthesis or assimilation of volatile fatty acids. All of them displayed evident inhibition profiles to biofilm formation. Notably, the combination of these three inhibitors possessed a remarkable ability to block the development of a multi-species biofilm with lower concentrations, suggesting an enhanced effect with their simultaneous use. This study demonstrates that targeting cellular metabolism is an effective way to inhibit biofilm formation derived from multi-species.

scholarly journals Microtitre Plate Assay for the Quantification of Biofilm Formation by Pathogenic Leptospira

2017 ◽  
Vol 12 (2) ◽  
pp. 146-153 ◽  
Author(s):  
Kasing Apun ◽  
Chai Fung Pui ◽  
Jennifer Jalan ◽  
Lesley Maurice Bi ◽  
Lela Su`ut ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Microtitre Plate ◽  
Pathogenic Leptospira ◽  
Plate Assay ◽  
Microtitre Plate Assay

A novel bacterium involved in the degradation of 2-methylindole isolated from sediment of Inner Deep Bay of Hong Kong

2015 ◽  
Vol 1 (1) ◽  
pp. 52 ◽  
Author(s):  
Karen Choi-Wan Yip ◽  
Ji-Dong GU Gu
Keyword(s):  
Hong Kong ◽  
16S Rdna ◽  
Carbon Sources ◽  
Capric Acid ◽  
Optimal Growth ◽  
Rrna Gene ◽  
Aerobic Bacterium ◽  
Biochemical Tests ◽  
Novel Bacterium ◽  
Deep Bay

A bacterial strain, designated as MPKc, was isolated from the mudflat sediment of Mai Po Inner Deep Bay of Hong Kong Mai Po Nature Reserve by enrichment culturing with 2-methylindole as the sole source of carbon and energy. The microorganism was a Gram-negative, rod-shaped (0.4–0.6 μm × 1.0–2.2 μm) and aerobic bacterium. Phylogenetic analysis based on the 16S rRNA gene sequences indicated that strain MPKc should be assigned as a novel bacterium, at least, at the species level. The 16S rDNA sequence most similiar to that of strain MPKc was Azoarcus evansii (94%) from available 16S rDNA sequences of the GenBank, indicating that strain MPKc was a member of the β-subclass of the Proteobacteria. Biochemical tests showed that strain MPKc was able to reduce nitrate to nitrogen. Carbon sources utilized by this strain included adipic acid, malate, citrate and phenylacetic acid although it only grew weakly on glucose, arabinose, mannose, mannitol, N-acetyl-glucosamine, maltose and gluconate. Strain MPKc showed no growth on capric acid. Its optimal growth occurred at 30°C, pH 6.5–7.5 and salinity 5–10‰. Strain MPKc was capable of degrading 80 μM 2-methylindole in 7 days under aerobic conditions. The possible chemical pathway for 2-methylindole degradation is through oxidation at 3-position or/and 2-position of the pyrrole ring.

scholarly journals Thiorhodococcus mannitoliphagus sp. nov., a purple sulfur bacterium from the White Sea

2006 ◽  
Vol 56 (8) ◽  
pp. 1945-1951 ◽  
Author(s):  
Sandra Rabold ◽  
Vladimir M. Gorlenko ◽  
Johannes F. Imhoff
Keyword(s):  
White Sea ◽  
Sequence Similarity ◽  
Carbon Sources ◽  
Cell Diameter ◽  
Rrna Gene ◽  
Sulfur Source ◽  
The White Sea ◽  
Dna Base Composition ◽  
Optimal Salinity ◽  
Purple Sulfur Bacterium

A novel purple sulfur bacterium, strain WST, was isolated from a microbial mat from an estuary of the White Sea. Individual cells are coccoid shaped, motile by flagella and do not contain gas vesicles. The mean cell diameter is 1.85 μm (range 1.5–2.0 μm). Cell suspensions exhibit a purple–violet colour. They contain bacteriochlorophyll a and carotenoids of the rhodopinal series as photosynthetic pigments. The novel bacterium is an anoxygenic photoautotroph, using sulfide, thiosulfate, sulfite and elemental sulfur as electron donors for photosynthesis and is capable of photoassimilating several organic carbon sources in the presence of carbonate and a reduced sulfur source (sulfide and/or thiosulfate). Sulfur globules, formed during oxidation of sulfide, are stored transiently inside the cells. Optimal salinity and pH for growth are at 0.5–2.0 % NaCl and pH 7.0–7.5. The DNA base composition of strain WST is 61.8 mol% G+C. 16S rRNA gene sequence analysis showed that the new isolate belongs to the genus Thiorhodococcus, with Thiorhodococcus minor CE2203T as the nearest relative (sequence similarity of 97.3 %). Several distinct differences from described species necessitate the description of a novel species. Thiorhodococcus mannitoliphagus sp. nov. is the proposed name, with strain WST (=ATCC BAA-1228T=VKM B-2393T) as the type strain.

scholarly journals Aquabacterium olei sp. nov., an oil-degrading bacterium isolated from oil-contaminated soil

2015 ◽  
Vol 65 (Pt_10) ◽  
pp. 3597-3602 ◽  
Author(s):  
Van Hong Thi Pham ◽  
Seung-Woo Jeong ◽  
Jaisoo Kim
Keyword(s):  
Contaminated Soil ◽  
Sequence Similarity ◽  
Phylogenetic Analyses ◽  
Carbon Sources ◽  
Optimal Growth ◽  
Rrna Gene ◽  
16S Rrna Gene Sequences ◽  
Degrading Bacterium ◽  
Genotypic Characteristics ◽  
Type Strains

Strain NHI-1T is a Gram-negative, motile, non-spore-forming bacterium isolated from oil-contaminated soil in South Korea. The strain was able to grow by using gasoline, diesel and kerosene as energy and carbon sources. After incubation for 14 days, cells (1 g l− 1) degraded approximately 58 % of oil present at concentration of 1500 p.p.m. at pH 8 and 28 °C. Strain NHI-1T grew well under aerobic conditions, with optimal growth at pH 7–9 and 28 °C–37 °C but grew poorly in the presence of ≥ 0.5 % NaCl. Phylogenetic analyses based on 16S rRNA gene sequences indicated that the closest relatives of strain NHI-1T were Aquabacterium fontiphilum CS-6T (97.96 % sequence similarity), Aquabacterium parvum B6T (96.39 %), Aquabacterium commune B8T (95.76 %), Aquabacterium limnoticum ABP-4T (95.72 %) and Aquabacterium citratiphilum B4T (95.25 %). DNA–DNA relatedness was 41–53 % between strain NHI-1T and its closest type strains. The major fatty acids present in strain NHI-1T were summed feature 3 (C16 : 1ω7c/C16 : 1ω6c, 44.5 %), summed feature 8 (C18 : 1ω7c/C18 : 1ω6c, 21.5 %) and C16 : 0 (16.2 %), and the predominant polar lipids were phosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine, diphosphatidylglycerol and uncharacterized aminophospholipids. Strain NHI-1T was distinguishable from other members of genus Aquabacterium based on phenotypic, chemotaxonomic and genotypic characteristics. Therefore, strain NHI-1T represents a novel species of the genus Aquabacterium for which the name Aquabacterium olei sp. nov. is proposed. The type strain is NHI-1T ( = KEMB 9005-082T =  KACC 18244T = NBRC 110486T).

scholarly journals Characterization of Polyhydroxyalkanoates Produced by Contaminated Soil Bacteria using Wastewater and Glucose as Carbon Sources

2015 ◽  
Vol 14 (9) ◽  
pp. 1605-1611
Author(s):  
S Munir ◽  
N Jamil
Keyword(s):  
Carbon Source ◽  
Contaminated Soil ◽  
Volatile Fatty Acids ◽  
Carbon Sources ◽  
Bacterial Strains ◽  
Biochemical Tests ◽  
Different Types ◽  
Pha Production ◽  
Different Sources

Purpose: To isolate polyhydroxyalkanoates (PHA)-producing bacterial strains from contaminated soil using industrial wastewater and glucose as carbon soured by Macrogen sequencing. Two different sources, namely, glucose and wastewater were used to ces.Methods: The strains were isolated and identified as Pseudomonas, Bacillus, Enterobacter, Exiguobacterium and Stenotrophomonas using biochemical tests and further confirmevaluate and  compare the use of wastewater as a carbon source for PHA production. The biomass obtained was analyzed by Fourier transform infra-red (FTIR) to identify the presence of PHA in it. Afterwards, PHA extraction was carried out and then gas chromatography (GC) performed to identify PHA monomers.Results: Utilization of glucose resulted in the production of PHB, while wastewater yielded copolymers poly-3 hydroxybutyrate-co-3hydroxyvalerate P(3HB-co-3HV) due to its content of volatile fatty acids such as acetic acid, propionic acid and butyric acid, which led to the production of different types of polymers. The maximum PHA production was 41 ± 0.22 % obtained for Stenotrophomonas (SM03) using 2 % glucose as carbon source while for wastewater, maximum production was achieved by the Pseudomonas strain (SM01).Conclusion: Wastewater is produced in large quantities daily during various activities and therefore can be used as a cheap carbon source for the production of valuable products such as PHA.Keywords: Polyhydroxyalkanoates, Wastewater, Glucose, Pseudomonas strain, Stenotrophomonas

scholarly journals Description of Thermogemmatispora carboxidivorans sp. nov., a carbon-monoxide-oxidizing member of the class Ktedonobacteria isolated from a geothermally heated biofilm, and analysis of carbon monoxide oxidation by members of the class Ktedonobacteria

2014 ◽  
Vol 64 (Pt_4) ◽  
pp. 1244-1251 ◽  
Author(s):  
C. E. King ◽  
G. M. King
Keyword(s):  
Carbon Monoxide ◽  
Type Species ◽  
Phylogenetic Analyses ◽  
Carbon Sources ◽  
Optimal Growth ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
Orange Pigment ◽  
Content Type ◽  
Link Type

A thermophilic, aerobic, Gram-stain-positive bacterium (strain PM5T), which formed mycelia of irregularly branched filaments and produced multiple exospores per cell, was isolated from a geothermally heated biofilm. Strain PM5T grew at 40–65 °C and pH 4.1–8.0, with optimal growth at 55 °C and pH 6.0. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain PM5T belonged to the class Ktedonobacteria , and was related most closely to Thermogemmatispora onikobensis ONI-1T (97.7 % similarity) and Thermogemmatispora foliorum ONI-5T (96.1 %). Morphological features and fatty acid profiles (major fatty acids: iso-C17 : 0, iso-C19 : 0 and 12,17-dimethyl C18 : 0) supported the affiliation of strain PM5T to the genus Thermogemmatispora . Strain PM5T oxidized carbon monoxide [CO; 10±1 nmol h−1 (mg protein)−1], but did not grow with CO as a sole carbon and energy source. Results from analyses of related strains indicated that the capacity for CO uptake occurred commonly among the members of the class Ktedonobacteria ; 13 of 14 strains tested consumed CO or harboured coxL genes that potentially enabled CO oxidation. The results of DNA–DNA hybridization and physiological and biochemical tests allowed the genotypic and phenotypic differentiation of strain PM5T from the two recognized species of the genus Thermogemmatispora . Strain PM5T differed from Thermogemmatispora onikobensis ONI-1T in its production of orange pigment, lower temperature optimum, hydrolysis of casein and starch, inability to grow with mannitol, xylose or rhamnose as sole carbon sources, and utilization of organic acids and amino acids. Strain PM5T is therefore considered to represent a novel species, for which the name Thermogemmatispora carboxidivorans sp. nov. is proposed. The type strain is PM5T ( = DSM 45816T = ATCC BAA-2534T).

scholarly journals Tropicibacter multivorans sp. nov., an aerobic alphaproteobacterium isolated from surface seawater

2012 ◽  
Vol 62 (Pt_4) ◽  
pp. 844-848 ◽  
Author(s):  
Teresa Lucena ◽  
María J. Pujalte ◽  
María A. Ruvira ◽  
Esperanza Garay ◽  
M. Carmen Macián ◽  
...  
Keyword(s):  
Type Species ◽  
Sequence Similarity ◽  
Phylogenetic Analyses ◽  
Carbon Sources ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
Phenotypic Data ◽  
Content Type ◽  
Link Type ◽  
The Family

Strain MD5T, an aerobic marine alphaproteobacterium, was isolated from Mediterranean seawater at Malvarrosa beach, Valencia, Spain. The strain was characterized in a polyphasic study and was placed phylogenetically within the Roseobacter clade in the family Rhodobacteraceae . Phylogenetic analysis based on 16S rRNA gene sequences showed that strain MD5T is related to Tropicibacter naphthalenivorans C02T, Phaeobacter inhibens T5T, P. gallaeciensis BS107T and P. daeponensis TF-218T, with 96.9, 96.2, 96.1 and 96.1 % sequence similarity, respectively. Phylogenetic analyses also showed that strain MD5T forms a stable clade only with T. naphthalenivorans C02T. Strain MD5T requires Na+ plus a divalent cation (either Mg2+ or Ca2+) to grow, does not reduce nitrate to nitrite and uses a large number of carbohydrates as sole carbon sources. It is positive for β-galactosidase and urease activities and aesculin hydrolysis. Enzyme activities displayed in the API ZYM strip were alkaline phosphatase, leucine arylamidase, acid phosphatase and α-glucosidase. Major cellular fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c; 70.9 %) and C16 : 0 (8.2 %). The results of physiological and biochemical tests allowed clear phenotypic differentiation of this isolate from the only described species of the genus Tropicibacter . It is evident from the genotypic and phenotypic data obtained that the strain should be classified in a novel species in the genus Tropicibacter . The name Tropicibacter multivorans sp. nov. is proposed, with the type strain MD5T ( = CECT 7557T  = KCTC 23350T).

Georgenia soli sp. nov., isolated from iron-ore-contaminated soil in India

2010 ◽  
Vol 60 (5) ◽  
pp. 1027-1030 ◽  
Author(s):  
P. Kämpfer ◽  
A. B. Arun ◽  
H.-J. Busse ◽  
S. Langer ◽  
C.-C. Young ◽  
...  
Keyword(s):  
Contaminated Soil ◽  
Iron Ore ◽  
Gene Sequence ◽  
Novel Species ◽  
Sequence Similarity ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
Polar Lipid Profile ◽  
Phenotypic Differentiation ◽  
Physiological And Biochemical

A Gram-positive actinobacterium (CC-NMPT-T3T) was isolated from iron-ore-contaminated soil near New Mangalore Port, Karnataka, India. Based on 16S rRNA gene sequence similarity studies, strain CC-NMPT-T3T belongs to the genus Georgenia and is most closely related to Georgenia muralis (98.6 %), Georgenia ruanii (97.4 %) and Georgenia thermotolerans (97.4 %). The peptidoglycan is of the type A4α l-Lys←l-Glu. The predominant isoprenoid quinone is menaquinone MK-8(H4) and the polar lipid profile is composed of the predominant compound diphosphatidylglycerol, moderate amounts of a phosphatidylinositol-mannoside, phosphatidylinositol and minor amounts of another phosphatidylmannoside and phosphatidylglycerol. The major fatty acids of strain CC-NMPT-T3T are anteiso-C15 : 0 and iso-C15 : 0. The results of DNA–DNA hybridizations, and physiological and biochemical tests allowed a clear phenotypic differentiation of strain CC-NMPT-T3T from all other Georgenia species. Strain CC-NMPT-T3T represents a novel species, for which the name Georgenia soli sp. nov. is proposed. The type strain is CC-NMPT-T3T (=DSM 21838T=CCM 7658T).

scholarly journals Antibiogram and Biofilm Formation among Carbapenem Resistant Klebsiella pneumoniae

2019 ◽  
Vol 6 ◽  
pp. 63-69
Author(s):  
Roshani Nhuchhen Pradhan ◽  
Surendra Kumar Madhup ◽  
Shyam Prasad Pant
Keyword(s):  
Klebsiella Pneumoniae ◽  
Congo Red ◽  
Biofilm Formation ◽  
Microtitre Plate ◽  
Sensitivity Testing ◽  
Clinical Specimens ◽  
Plate Assay ◽  
Carbapenem Resistant ◽  
Microtitre Plate Assay ◽  
Agar Method

Objectives: This cross-sectional study was designed to detect the carbapenemase producing K. pneumoniae along with biofilm producers from different clinical specimens and to compare antibiotic susceptibility pattern of biofilm producing carbapenem resistant Klebsiella pneumoniae and biofilm non-producing carbapenem resistant Klebsiella pneumoniae. Methods: A total of 1475 non-repetitive clinical samples were included on this study. Antibiotic Sensitivity Testing (AST), Modified Hodge Test (MHT) and Modified Carbapenem inactivation method (mCIM) were performed for detection of carbapenemase production and Congo red agar method (CRA) along with Microtitre plate method were performed for detecting biofilm production. Results: Among the clinical specimens cultured, growth positivity was 62.71%. E. coli was most predominant organism followed by K. pneumoniae (17.89%). Among the 110 K. pneumoniae, 57 were found to be carbapenemase producer. Majority of the carbapenemase producing K. pneumoniae were isolated from sputum (45.61%), in the specimen collected from age group 61-70 (28.07%) and in out-patient department (50.88%). Similarly, 65.45% K. pneumoniae out of 110 were found to be biofilm producer by Congo red agar method while among those 72, 73.59% isolates were found to be quantitatively biofilm producer in Microtitre plate assay. Out of 57 carbapenemase producer, 35.08% were strongly biofilm producer while among 53 carbapenemase non-producer 30.18% were strongly biofilm producer from Congo red agar method. Moreover, Microtitre plate assay evidenced that, out of 57 carbapenemase producer, 40.35% were highly biofilm producing and among the 15 carbapenemase nonproducer 66.66% were highly biofilm producer.  Conclusion: Biofilm formation is highly prevalent with varying degree of resistance among different antibiotics including carbapenems that further augments antibiotic resistance. The study showed carbapenemase producers are stronger biofilm producer than the non-carbapenemase producer. Therefore, it is recommended to identify biofilm formation among carbapenemase producers for effective choice of antibiotics.

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