Isolation and identification of the same type LMG1242 of biosurfactant-producing strain of Pseudomonas aeruginosa from different oil contaminated soils

Emulsification is a major rate limiting step in any biological conversion of heavy hydrocarbons. Here, thirty bacterial strains were isolated from seven native consortia and screened for biosurfactant-producing activities which could enhance emulsification of heavy oils. The consortia were obtained by sampling from oil contaminated soils of different petroleum refineries of Iran. The oil spreading test, drop collapse test, emulsification index (E24) and surface tension measurements were used to evaluate the biosurfactant producing activities of the strains. A total number of 5 strains out of 30 were finally selected as the best biosurfactant-producing bacteria. The clear zone diameters in their oil spreading test were 4 to 5 cm, and E24 were 48 to 92%. The selected strains also properly lowered the surface tensions of the supernatants to 23.5-32 mN/m when grown on heavy diesel. All the selected strains were identified by 16S rRNA sequencing analysis as Pseudomonas aeruginosa LMG1242 (99.9% homology) which is a newly isolated type in oil polluted soils. The results suggest P. aeruginosa LMG1242 as a predominant and highly active biosurfactant producing bacterium which could be further evaluated in petroleum bioremediation and bioprocessing applications.

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Интенсификация биоремедиации нефтезагрязненных почв с использованием природного консорциума микроорганизмов

SCIENCE & TECHNOLOGIES OIL AND OIL PRODUCTS PIPELINE TRANSPORTATION ◽

10.28999/2541-9595-2021-11-5-582-589 ◽

2021 ◽

pp. 582-589

Author(s):

Kalamkas B. Oralbayeva ◽

Aliya K. Yernazarova ◽

Medet E. Kundybayev ◽

Spandiar A. Seitkulov ◽

Aida S. Kistaubayeva

Keyword(s):

Pseudomonas Aeruginosa ◽

Contaminated Soil ◽

Contaminated Soils ◽

Climatic Conditions ◽

Mineral Fertilizers ◽

Pseudomonas Sp ◽

Bacterial Strains ◽

Biological Product ◽

Metabolic Potential ◽

Alcaligenes Denitrificans

Currently biological methods of cleaning oily soils significantly developed as the most accessible and effective. In their basis – the use of hydrocarbon-oxidizing microorganisms (HOM) metabolic potential. At the same time, as practice shows, existing preparations containing HOM are not effective enough in extreme soil-climatic conditions, in this connection the search for indigenous strains of oil-oxidizing bacteria and the development of new biological preparations are of current interest. The aim of this work is to study the effectiveness of HOM strains for restoring the biological activity of oil-contaminated soil. A study of the microflora of unpolluted and oil-contaminated soil taken from the experimental site of the “Shymkent” oil pumping station of “KazTransOil” JSC was carried out: the total number of microorganisms was 210·106cells/g and 130·106 cells/g, respectively. From 16 НOM strains isolated from oil-contaminated soil, 3 bacterial strains were selected to create an association and use as a biological product-oil destructor – P12, P16, A8. Based on the results of genotyping, strains P12 and P16 correspond to the species Pseudomonas aeruginosa, strain A8 belongs to the species Alcaligenes denitrificans. The associations of Pseudomonas sp. P16, Pseudomonas sp. P12 and Alcaligenes sp. A8 showed better growth in comparison with monocultures on a nutrient medium with hydrocarbons. Bioremediation of artificially oil polluted site process, which carried out for 120 days using a biological product created on the basis of the association of strains P12, P16 and A8, decreased the content of hydrocarbons 3.4 times, and with additional application of mineral fertilizers (ammophos) – 4.6 times. Thus, proved the expediency of using HOM strains P12, P16, A8 for the purpose of bioremediation of oil-contaminated soils. The results of the study can be used to improve economical and environmental efficiency of work when carrying out measures to eliminate the consequences of oil spills on the ground. Биологические методы очистки нефтесодержащих почв в настоящее время имеют существенное развитие как наиболее доступные и эффективные. В их основе – использование метаболического потенциала углеводородокисляющих микроорганизмов (УОМ). При этом, как показывает практика, существующие препараты, содержащие УОМ, недостаточно эффективны в экстремальных почвенно-климатических условиях, в связи с чем актуальными являются вопросы поиска аборигенных штаммов нефтеокисляющих бактерий и разработка новых биопрепаратов. Цель работы – исследование эффективности штаммов УОМ для восстановления биологической активности нефтезагрязненного почвогрунта. Проведено изучение микрофлоры незагрязненной и нефтезагрязненной почвы, отобранной с экспериментального участка ГНПС «Шымкент» АО «КазТрансОйл»: общее количество микроорганизмов составило 210·106 кл/г и 130·106кл/г, соответственно. Из выделенных из нефтезагрязненной почвы 16 штаммов УОМ были отобраны три штамма бактерий для создания ассоциации и применения в качестве биопрепарата-нефтедеструктора – Р12, Р16, А8. По итогам генотипирования установлено, что штаммы Р12 и Р16 соответствуют виду Pseudomonas aeruginosa, штамм А8 относится к виду Alcaligenes denitrificans. Установлено, что в составе ассоциаций штаммы Pseudomonas sp. P16, Pseudomonas sp. P12 и Alcaligenes sp. A8. показали лучший по сравнению с монокультурами рост на питательной среде с углеводородами. В процессе биоремедиации участка с искусственным нефтезагрязнением, проводимой в течение 120 суток с использованием биопрепарата, созданного на основе ассоциации штаммов P12, P16 и A8, содержание углеводородов снизилось в 3,4 раза, а при дополнительном внесении минеральных удобрений (аммофоса) – в 4,6 раза. Таким образом, доказана целесообразность использования штаммов УОМ Р12, Р16, А8 с целью биоремедиации нефтезагрязненных почв. Результаты исследования могут быть использованы для повышения экономической и экологической эффективности работ при проведении мероприятий по ликвидации последствий разливов нефти на почве.

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Characterizations of novel pesticide-degrading bacterial strains from industrial wastes found in the industrial cities of Pakistan and their biodegradation potential

PeerJ ◽

10.7717/peerj.12211 ◽

2021 ◽

Vol 9 ◽

pp. e12211

Author(s):

Noreen Asim ◽

Mahreen Hassan ◽

Farheen Shafique ◽

Maham Ali ◽

Hina Nayab ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Enrichment Culture ◽

Culture Technique ◽

Industrial Wastes ◽

Bacterial Strains ◽

Isolation And Identification ◽

16S Rrna Analysis ◽

Degrading Bacteria ◽

Considerable Degradation ◽

Different Strains

Background Lack of infrastructure for disposal of effluents in industries leads to severe pollution of natural resources in developing countries. These pollutants accompanied by solid waste are equally hazardous to biological growth. Natural attenuation of these pollutants was evidenced that involved degradation by native microbial communities. The current study encompasses the isolation of pesticide-degrading bacteria from the vicinity of pesticide manufacturing industries. Methods The isolation and identification of biodegrading microbes was done. An enrichment culture technique was used to isolate the selected pesticide-degrading bacteria from industrial waste. Results Around 20 different strains were isolated, among which six isolates showed significant pesticide biodegrading activity. After 16S rRNA analysis, two isolated bacteria were identified as Acinetobacter baumannii (5B) and Acidothiobacillus ferroxidans, and the remaining four were identified as different strains of Pseudomonas aeruginosa (1A, 2B, 3C, 4D). Phylogenetic analysis confirmed their evolution from a common ancestor. All strains showed distinctive degradation ability up to 36 hours. The Pseudomonas aeruginosa strains 1A and 4D showed highest degradation percentage of about 80% for DDT, and P. aeruginosa strain 3C showed highest degradation percentage, i.e., 78% for aldrin whilst in the case of malathion, A. baumannii and A. ferroxidans have shown considerable degradation percentages of 53% and 54%, respectively. Overall, the degradation trend showed that all the selected strains can utilize the given pesticides as sole carbon energy sources even at a concentration of 50 mg/mL. Conclusion This study provided strong evidence for utilizing these strains to remove persistent residual pesticide; thus, it gives potential for soil treatment and restoration.

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Isolation and identification of chromium-tolerant bacterial strains and their potential to promote plant growth

E3S Web of Conferences ◽

10.1051/e3sconf/20199601005 ◽

2019 ◽

Vol 96 ◽

pp. 01005

Author(s):

Muhammad Arshad ◽

Anza Javaid ◽

Maria Manzoor ◽

Kiran Hina ◽

Muhammad Arif Ali ◽

...

Keyword(s):

Plant Growth ◽

Contaminated Soils ◽

Rrna Gene ◽

Bacterial Strains ◽

Isolation And Identification ◽

Pseudomonas Plecoglossicida ◽

Plant Growth Promoting ◽

Promote Plant Growth ◽

Growth Promoting

Remediation and management of Chromium (Cr) contaminated soils for safe usage has been a serious challenge. The aim of present work was to isolate and identify Cr tolerant bacterial strains and assessing their plant growth promoting potential under controlled conditions. Soil samples were collected from Gujrat District, Pakistan, and analyzed for Cr content and bacteria isolation. In-vitro screening was done for chromium tolerance and plant growth-promoting (PGP) abilities. The tolerant isolates which also exhibited PGP abilities were used as inoculants in germination and pot trial experiment. The isolates A5 and A6, identified as Pseudomonas plecoglossicida and Staphylococci saprophyticus through 16S rRNA gene sequencing, were found to be tolerant up to 700 mg L-1 of Cr (VI). Both were effective in solubilizing phosphate but only A5 (Pseudomonas plecoglossicida) was able to produce indole acetic acid. A5 also increased the percentage of seed germination from 17 to 46% and spinach plant’s biomass by 44% with respect to the control. Keeping in view the results obtained, A5 (Pseudomonas plecoglossicida) appeared as the best species that was able to tolerate Cr stress and promote plant growth. Further, it can be developed as a bio-inoculant for non-food agricultural applications for remediation of Cr contaminated soils.

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Pseudomonas aeruginosa: an emerging nosocomial trouble in veterinary

Revista Mvz Córdoba ◽

10.21897/rmvz.9 ◽

2015 ◽

pp. 4937-4946 ◽

Cited By ~ 5

Author(s):

Yuly Bernal-Rosas ◽

Karen Osorio-Muñoz ◽

Orlando Torres-García

Keyword(s):

Pseudomonas Aeruginosa ◽

Multidrug Resistant ◽

Diffusion Method ◽

Bacterial Strains ◽

Isolation And Identification ◽

Disk Diffusion Method ◽

Clinical Specimens ◽

Mueller Hinton ◽

Mueller Hinton Agar ◽

Intermediate Response

ABSTRACT Objective. The goal of this study was to evaluate the susceptibility pattern of isolates P. aeruginosa from veterinary clinical centers in Bogotá, D.C., to some commonly used antibiotics in clinical. Materials and methods. Bacteriological standard protocols were used for the isolation and identification of bacterial strains. To evaluate the antimicrobial susceptibility of the isolates, to commonly used antibiotics, was performed the Kirby-Bauer agar-disk diffusion method on Mueller-Hinton agar. Results. A total of 160 samples was taken from clinical specimens and the environment in different veterinary clinics. Out of these samples, 89 (55.6%) were gram-negative strains, of which ten strains of P. aeruginosa were isolated (11.2%). All strains were resistant to Cefazolin, Lincomycin, Cephalothin, Ampicillin, Clindamycin, Sulfamethoxazole-Trimethoprim and Chloramphenicol while some isolates exhibited either resistance or an intermediate response to Amikacin (30%), Gentamicin (30%), Tobramycin (10%), Ciprofloxacin (20%), Ceftazidime (30%), Erythromycin (100%), Tetracycline (100%), Imipenem (10%), Meropenem (90%) and Bacitracin (90%). Conclusions. The results demonstrate that the acquired antimicrobial resistances of P. aeruginosa strains depend on antibiotic protocols applied. As observed in human hospitals, Pseudomonas aeruginosa is acting as one of the multidrug-resistant microorganisms of veterinary clinical relevance.

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Isolation and identification of IAA producing endosymbiotic bacteria from Gracillaria corticata (J. Agardh)

International Journal of Bioassays ◽

10.21746/ijbio.2016.12.0012 ◽

2016 ◽

Vol 5 (12) ◽

pp. 5179

Author(s):

Ilahi Shaik* ◽

P. Janakiram ◽

Sujatha L. ◽

Sushma Chandra

Keyword(s):

Acetic Acid ◽

Culture Filtrate ◽

Indole Acetic Acid ◽

Shoot Growth ◽

High Salinity ◽

Saline Soils ◽

Bacterial Strains ◽

Isolation And Identification ◽

Endosymbiotic Bacteria ◽

Root And Shoot Growth

Indole acetic acid is a natural phytohormone which influence the root and shoot growth of the plants. Six (GM1-GM6) endosymbiotic bacteria are isolated from Gracilaria corticata and screened for the production of IAA out of six, three bacterial strains GM3, GM5 and GM6 produced significant amount of IAA 102.4 µg/ml 89.40 µg/ml 109.43 µg/ml respectively. Presence of IAA in culture filtrate of the above strains is further analyzed and confirmed by TLC. As these bacterial strains, able to tolerate the high salinity these can be effectively used as PGR to increase the crop yield in saline soils.

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Specific and rapid reverse assaying protocol for detection and antimicrobial susceptibility testing of Pseudomonas aeruginosa based on dual molecular recognition

Scientific Reports ◽

10.1038/s41598-021-90619-3 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Yong He ◽

Hang Zhao ◽

Yuanwen Liu ◽

He Zhou

Keyword(s):

Pseudomonas Aeruginosa ◽

Antimicrobial Susceptibility ◽

Magnetic Particles ◽

Susceptibility Testing ◽

Fluorescein Isothiocyanate ◽

High Specificity ◽

Antimicrobial Susceptibility Testing ◽

Tail Fiber ◽

Isolation And Identification ◽

Magainin Ii

AbstractThe worldwide emergence and spread of antimicrobial resistance is accelerated by irrational administration and use of empiric antibiotics. A key point to the crisis is a lack of rapid diagnostic protocols for antimicrobial susceptibility testing (AST), which is crucial for a timely and rational antibiotic prescription. Here, a recombinant bacteriophage tail fiber protein (TFP) was functionalized on magnetic particles to specifically capture Pseudomonas aeruginosa, while fluorescein isothiocyanate-labeled-magainin II was utilized as the indicator. For solving the magnetic particles’ blocking effects, a reverse assaying protocol based on TFP recognition was developed to investigate the feasibility of detection and AST of P. aeruginosa. P. aeruginosa can be rapidly, sensitively and specifically detected within 1.5 h with a linear range of 1.0 × 102 to 1.0 × 106 colony forming units (CFU)⋅mL−1 and a detection limit of 3.3 × 10 CFU⋅mL−1. Subsequently, AST results, which were consistent with broth dilution results, can be obtained within 3.5 h. Due to the high specificity of the TFP, AST can actually be conducted without the need for bacterial isolation and identification. Based on the proof-of-principle work, the detection and AST of other pathogens can be extended by expressing the TFPs of their bacteriophages.

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Microbiome of Odontogenic Abscesses

Microorganisms ◽

10.3390/microorganisms9061307 ◽

2021 ◽

Vol 9 (6) ◽

pp. 1307

Author(s):

Sebastian Böttger ◽

Silke Zechel-Gran ◽

Daniel Schmermund ◽

Philipp Streckbein ◽

Jan-Falco Wilbrand ◽

...

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Oral Microbiome ◽

Minor Role ◽

Rrna Gene ◽

Sequencing Analysis ◽

Bacterial Strains ◽

16S Rrna Gene Analysis ◽

Odontogenic Infections ◽

Odontogenic Abscess

Severe odontogenic abscesses are regularly caused by bacteria of the physiological oral microbiome. However, the culture of these bacteria is often prone to errors and sometimes does not result in any bacterial growth. Furthermore, various authors found completely different bacterial spectra in odontogenic abscesses. Experimental 16S rRNA gene next-generation sequencing analysis was used to identify the microbiome of the saliva and the pus in patients with a severe odontogenic infection. The microbiome of the saliva and the pus was determined for 50 patients with a severe odontogenic abscess. Perimandibular and submandibular abscesses were the most commonly observed diseases at 15 (30%) patients each. Polymicrobial infections were observed in 48 (96%) cases, while the picture of a mono-infection only occurred twice (4%). On average, 31.44 (±12.09) bacterial genera were detected in the pus and 41.32 (±9.00) in the saliva. In most cases, a predominantly anaerobic bacterial spectrum was found in the pus, while saliva showed a similar oral microbiome to healthy individuals. In the majority of cases, odontogenic infections are polymicrobial. Our results indicate that these are mainly caused by anaerobic bacterial strains and that aerobic and facultative anaerobe bacteria seem to play a more minor role than previously described by other authors. The 16S rRNA gene analysis detects significantly more bacteria than conventional methods and molecular methods should therefore become a part of routine diagnostics in medical microbiology.

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Role of Two Plant Growth-Promoting Bacteria in Remediating Cadmium-Contaminated Soil Combined with Miscanthus floridulus (Lab.)

Plants ◽

10.3390/plants10050912 ◽

2021 ◽

Vol 10 (5) ◽

pp. 912

Author(s):

Shuming Liu ◽

Hongmei Liu ◽

Rui Chen ◽

Yong Ma ◽

Bo Yang ◽

...

Keyword(s):

Plant Growth ◽

Contaminated Soils ◽

Translocation Factor ◽

Plant Growth Promoting Bacteria ◽

Bacterial Strains ◽

Pgp Traits ◽

Cd Tolerance ◽

Plant Growth Promoting ◽

Growth Promoting ◽

Cd Translocation

Miscanthus spp. are energy plants and excellent candidates for phytoremediation approaches of metal(loid)s-contaminated soils, especially when combined with plant growth-promoting bacteria. Forty-one bacterial strains were isolated from the rhizosphere soils and roots tissue of five dominant plants (Artemisia argyi Levl., Gladiolus gandavensis Vaniot Houtt, Boehmeria nivea L., Veronica didyma Tenore, and Miscanthus floridulus Lab.) colonizing a cadmium (Cd)-contaminated mining area (Huayuan, Hunan, China). We subsequently tested their plant growth-promoting (PGP) traits (e.g., production of indole-3-acetic acid, siderophore, and 1-aminocyclopropane-1-carboxylate deaminase) and Cd tolerance. Among bacteria, two strains, Klebsiella michiganensis TS8 and Lelliottia jeotgali MR2, presented higher Cd tolerance and showed the best results regarding in vitro growth-promoting traits. In the subsequent pot experiments using soil spiked with 10 mg Cd·kg−1, we investigated the effects of TS8 and MR2 strains on soil Cd phytoremediation when combined with M. floridulus (Lab.). After sixty days of planting M. floridulus (Lab.), we found that TS8 increased plant height by 39.9%, dry weight of leaves by 99.1%, and the total Cd in the rhizosphere soil was reduced by 49.2%. Although MR2 had no significant effects on the efficiency of phytoremediation, it significantly enhanced the Cd translocation from the root to the aboveground tissues (translocation factor > 1). The combination of K. michiganensis TS8 and M. floridulus (Lab.) may be an effective method to remediate Cd-contaminated soils, while the inoculation of L. jeotgali MR2 may be used to enhance the phytoextraction potential of M. floridulus.

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Antimicrobial Activity of Cyclic-Monomeric and Dimeric Derivatives of the Snail-Derived Peptide Cm-p5 against Viral and Multidrug-Resistant Bacterial Strains

Biomolecules ◽

10.3390/biom11050745 ◽

2021 ◽

Vol 11 (5) ◽

pp. 745

Author(s):

Melaine González-García ◽

Fidel Morales-Vicente ◽

Erbio Díaz Pico ◽

Hilda Garay ◽

Daniel G. Rivera ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Antimicrobial Activity ◽

Antifungal Activity ◽

Multidrug Resistant ◽

Moderate Activity ◽

Dependent Manner ◽

Bacterial Strains ◽

Acinetobacter Baumanii ◽

Concentration Dependent Manner ◽

Conventional Antibiotics

Cm-p5 is a snail-derived antimicrobial peptide, which demonstrated antifungal activity against the pathogenic strains of Candida albicans. Previously we synthetized a cyclic monomer as well as a parallel and an antiparallel dimer of Cm-p5 with improved antifungal activity. Considering the alarming increase of microbial resistance to conventional antibiotics, here we evaluated the antimicrobial activity of these derivatives against multiresistant and problematic bacteria and against important viral agents. The three peptides showed a moderate activity against Pseudomonas aeruginosa, Klebsiella pneumoniae Extended Spectrum β-Lactamase (ESBL), and Streptococcus agalactiae, with MIC values > 100 µg/mL. They exerted a considerable activity with MIC values between 25–50 µg/mL against Acinetobacter baumanii and Enterococcus faecium. In addition, the two dimers showed a moderate activity against Pseudomonas aeruginosa PA14. The three Cm-p5 derivatives inhibited a virulent extracellular strain of Mycobacterium tuberculosis, in a dose-dependent manner. Moreover, they inhibited Herpes Simplex Virus 2 (HSV-2) infection in a concentration-dependent manner, but had no effect on infection by the Zika Virus (ZIKV) or pseudoparticles of Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2). At concentrations of >100 µg/mL, the three new Cm-p5 derivatives showed toxicity on different eukaryotic cells tested. Considering a certain cell toxicity but a potential interesting activity against the multiresistant strains of bacteria and HSV-2, our compounds require future structural optimization.

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