Application of pretreatments for the isolation of bioactive actinomycetes from marine sediments

Pisano, Michael A.; Sommer, Michael J.; Lopez, Madelyn M.

doi:10.1007/bf00253664

Cited by 49 publications

(33 citation statements)

References 8 publications

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“…Then, mortar and pestle were used to grind the samples into smaller particles to increase the efficiency of bacterial isolation. Phenol (1.5%, 30 min at 30°C) was used for pretreatment of soil samples according to Pisano et al (1986). The pretreated soil samples were diluted 1:10 (v/v) with sterile ¼ Ringer solution and serial dilution up to 10 -4 .…”

Section: Isolation Of Proteobacteriamentioning

confidence: 99%

Analysis of Antarctic proteobacteria by PCR fingerprinting and screening for antimicrobial secondary metabolites

et al. 2012

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ABSTRACT. Fifty-seven proteobacterium species were successfully isolated from soils of Barrientos Island of the Antarctic using 11 different isolation media. Analysis of 16S rDNA sequencing of these isolates showed that they belonged to eight different genera, namely Bradyrhizobium, Sphingomonas, Methylobacterium, Caulobacter, Paracoccus, Ralstonia, Rhizobium, and Staphylococcus. All isolates were studied for capability of producing antimicrobial and antifungal secondary metabolites using high-throughput screening models. Approximately 23 (13/57) and 2% (1/57) of isolates inhibited growth of Candida albicans ATCC 10231 T and Staphylococcus aureus ATCC 51650 T , respectively. These results indicated that proteobacterium species isolates from Antarctic could serve as potential source of useful bioactive metabolites. Enterobacterial repetitive intergenic consensus (ERIC)-PCR fingerprinting produced nine clusters and 13 single isolates, with a high D value of 0.9248. RAPD fingerprinting produced six clusters and 13 single isolates, with a relatively low D value of 0.7776. ERIC-PCR analysis proved to have better discrimination capability than RAPD analysis and generated better clustering for all proteobacterium species isolates. We conclude that ERIC-PCR is a robust, reliable and rapid molecular typing method for discriminating different genera of proteobacteria.

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Section: Isolation Of Proteobacteriamentioning

confidence: 99%

Analysis of Antarctic proteobacteria by PCR fingerprinting and screening for antimicrobial secondary metabolites

et al. 2012

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“…Sin duda, el pretratamiento de la muestra en calor fue eficaz para restringir el crecimiento de la flora acompañante, ya que en la mayoría de los cultivos se obtuvo colonias puras de actinomycetes. Estos resultados fueron similares a los obtenidos por Pisano et al (1986), quienes además del calor aplicaron tratamientos con fenol y posterior cultivo en agar quitina. En nuestro estudio, como resultado de ambos métodos se llegaron a aislar un total de 62 cepas de actinomycetes marinos cuyas características culturales se indican en la Tabla 2.…”

Section: Resultados Y Discusión Enriquecimiento Y Aislamiento De Actiunclassified

“…El protocolo de aislamiento que resultó más factible para obtener mayor número de cepas de actinomycetes marinos fue aquel en el que se realizó el tratamiento térmico húmedo y el cultivo en Agar Marino (75% de rendimiento). Este resultado difiere de otros trabajos, puesto que el Agar Marino muy útil generalmente en el aislamiento de heterótrofos marinos no es utilizado necesariamente para el aislamiento de actinomycetes, pero sí el Agar Caseína-Almidón, Agar Czapeck y otros medios enriquecidos, los cuales según otros autores (Pisano et al, 1986;Takizawa et al, 1993) tuvieron un óptimo rendimiento para la recuperación de actinomycetes marinos. En nuestro caso, el Agar Caseína-Almidón y el Agar Czapeck fueron ocasionalmente utilizados y tuvieron un rendimiento discreto.…”

Section: Resultados Y Discusión Enriquecimiento Y Aislamiento De Actiunclassified

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Actinomycetes bioactivos de sedimento marino de la costa central del Perú

et al. 2007

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En el presente trabajo evaluamos la actividad antibacteriana y antifúngica de actinomycetes marinos sobre patógenos de origen clínico. Asimismo, fueron evaluadas la capacidad de producir enzimas extracelulares como carbohidrasas, lipasas y proteasas. Los Actinomycetes fueron aislados de sedimentos colectados entre setiembre a diciembre del 2005 de las Bahías de Ancón (Lima) e Independencia (Ica) de 34 y 100 m de profundidad. El aislamiento se realizó en Agar Caseína - Almidón (ACA) y Agar Marino (AM) con adición de Cicloheximide (10 μg/mL). Las evaluaciones antimicrobianas fueron realizadas frente a bacterias patógenas antibiótico-multirresistentes y hongos de origen clínico; en tanto, para evaluar su actividad multienzimática se utilizaron sustratos poliméricos diversos. Se aislaron un total de 62 actinomycetes, de los cuales 31 (50%) mostraron actividad antibacteriana frente a Staphylococcus aureus, 36 (59%) frente a Pseudomonas aeruginosa y 23 (37%) a ambos patógenos. Las cepas de actinomycetes I-400A y M10-77 identificadas en cada caso como Streptomyces y Thermoactinomyces fueron las que exhibieron mayor actividad inhibitoria frente a P. aeruginosa y S. aureus respectivamente. Asimismo, 13 actinomycetes (20,97%) mostraron actividad antifúngica frente a cultivos de Candida albicans cepa 1511 y 17 (27,42%) frente a Candida albicans cepa 1511MIC; sin embargo, ningún actinomycete presentó actividad inhibitoria frente a Aspergillus niger, Aspergillus fumigatus y Trichophyton mentagrophytes. La mayoría de los actinomycetes mostraron tener actividad multienzimática capaz de hidrolizar compuestos poliméricos como el tween-80 (96%), la gelatina (95%), almidón (93%), lecitina (88%) y la caseína (74%). Extractos del compuesto activo obtenidos de la cepa M10-77 con etil acetato rindieron notable actividad inhibitoria contra S. aureus. Se concluye que el sedimento marino es fuente de Actinomycetes con gran capacidad de producir sustancias bioactivas de importancia médica-farmacológica e industrial.

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“…Acid treatment was conducted by the acidifying the samples to pH 2 were obtained for 3 hours. Heat-shock treatment was conducted by the heating the samples at 60  C for 4 hours (Pisano et al 1986). Treated samples were then inoculated onto starch agar medium (1% w/v starch, 0.4% w/v yeast extract, 0.2% w/v peptone, natural seawater and 2% w/v agar) and incubated for 4-8 weeks at room temperature.…”

Section: Isolation Of Actinomycetesmentioning

confidence: 99%

Marine Actinomycetes screening of Banten West Coast and their antibiotics purification

Sunaryanto¹,

Marwoto²

2010

Biodiversitas

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Sunaryanto R, Marwoto B (2010) Marine Actinomycetes screening of Banten West . Isolation and purification of active compounds produced by marine Actinomycetes has been carried out. Marine sediment samples were obtained from six different places at Anyer, Banten West Coast in October 20, 2007. Isolation was carried out using two methods pretreatments, acid treatment and heat shock treatment. A total of 29 Actinomycetes isolates were obtained from the various sediment samples collected, then tested for antimicrobial test against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC25923, Pseudomonas aeruginosa ATCC27853, Bacillus subtilis ATCC 66923, Candida albicans BIOMCC00122 and Aspergillus niger BIOMCC00134. Identification of potential isolate was carried out using 16S rRNA. Purification of active compound was carried out using silica gel column chromatography and preparative HPLC. Result of this research showed that isolate A11 produced the most active compound against Gram-positive and Gram-negative bacteria. Morphology and identification test using 16S rRNA gen showed that isolate A11 is Streptomyces sp. Production of active compound from isolate A11 used yeast peptone medium. The single peak of active compound was detected by HPLC and showed retention time on 8.35 min and maximum absorbance UV visible of antibiotic was 210 nm and 274.5 nm. Active purified compound showed inhibition activity to Gram-positive and Gram-negative bacteria. Minimum inhibitory concentration (MIC) to E. coli ATCC 25922 was 27 µg/mL, P. aeruginosa ATCC 27853 68.7 µg/mL, S. aureus ATCC 25923 80.2 µg/mL, and B. subtilis ATCC 66923 73.7 µg/mL.

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Application of pretreatments for the isolation of bioactive actinomycetes from marine sediments

Cited by 49 publications

References 8 publications

Analysis of Antarctic proteobacteria by PCR fingerprinting and screening for antimicrobial secondary metabolites

Analysis of Antarctic proteobacteria by PCR fingerprinting and screening for antimicrobial secondary metabolites

Actinomycetes bioactivos de sedimento marino de la costa central del Perú

Marine Actinomycetes screening of Banten West Coast and their antibiotics purification

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