Isolation of rare Genera of actinomycetes — antibiotic producers from soils using Aloe Arborescens juice

The search for new antibiotics is an urgent problem due to the spread of resistance to existing antibacterial drugs in pathogenic microorganisms. Actinomycetes are producers of a large number of antibiotics used in medicine. Most antibiotics are isolated from actinomycetes of the Streptomyces genus, while rare genera of actinomycetes can be the producers of new antibiotics.The aim of the study is to investigate the effect of the biological substances complex present in aloe juice on the growth stimulation of rare genera of actinomycetes.Material and methods. Objects: samples of sod-podzolic soil and chernozem. The standard method of sowing soil suspensions on oat agar and Gause medium No. 2 was used to isolate actinomycetes. Chemotaxonomic properties were determined using the methods of ascending thin-layer chromatography on a cellulose layer. The generic identity of cultures was determined using Bergey’s manual and materials comparing the composition of cell walls of actinobacteria. DNA PCR with standard 27f and 1492r primers, as well as Sanger sequencing, were performed to study genosystematic features. Antibiotic activity was determined against the test microorganisms: Staphylococcus aureus ИНА 00985 (FDA 209P), Staphylococcus aureus ИНА 00761 (MRSA), Staphylococcus aureus ИНА 00762 (УФ- 2), Micrococcus luteus ATCC 9341, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Saccharomyces cerevisiae ИНА 01042.Results. A total of 527 actinomycete cultures were isolated from samples of sod-podzolic soil and chernozem with the addition of aloe juice; their phylogenetic position was determined. The dominant actinomycetes in the studied soil samples are the representatives of the genus Streptomyces. Bacteria of the genus Micromonospora take the second place by the number of isolated cultures. Rare genera of actinomycetes have also been identified: Nonomuraea, Streptosporangium, Nocardia, Actinomadura, Actinocorallia, Pseudonocardia, Amycolatopsis, Saccharomonospora, Saccharopolyspora, Promicromonospora, Kribbella. It was determined that the isolated cultures possess antibiotic activity against test microorganisms.Conclusion. It is advisable to use aloe juice after subjecting the leaves to biostimulation to isolate actinomycetes from the soil and identify their biodiversity.

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Proliferation of Antibiotic-Producing Bacteria and Concomitant Antibiotic Production as the Basis for the Antibiotic Activity of Jordan's Red Soils

Applied and Environmental Microbiology ◽

10.1128/aem.00104-09 ◽

2009 ◽

Vol 75 (9) ◽

pp. 2735-2741 ◽

Cited By ~ 21

Author(s):

Joseph O. Falkinham ◽

Thomas E. Wall ◽

Justin R. Tanner ◽

Khaled Tawaha ◽

Feras Q. Alali ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Activity ◽

Incubation Period ◽

Micrococcus Luteus ◽

Antibiotic Production ◽

Antibiotic Activity ◽

Skin Infections ◽

High Antimicrobial Activity ◽

Methanol Extracts ◽

Red Soils

ABSTRACT Anecdotes, both historical and recent, recount the curing of skin infections, including diaper rash, by using red soils from the Hashemite Kingdom of Jordan. Following inoculation of red soils isolated from geographically separate areas of Jordan, Micrococcus luteus and Staphylococcus aureus were rapidly killed. Over the 3-week incubation period, the number of specific types of antibiotic-producing bacteria increased, and high antimicrobial activity (MIC, ∼10 μg/ml) was observed in methanol extracts of the inoculated red soils. Antibiotic-producing microorganisms whose numbers increased during incubation included actinomycetes, Lysobacter spp., and Bacillus spp. The actinomycetes produced actinomycin C2 and actinomycin C3. No myxobacteria or lytic bacteriophages with activity against either M. luteus or S. aureus were detected in either soil before or after inoculation and incubation. Although protozoa and amoebae were detected in the soils, the numbers were low and did not increase over the incubation period. These results suggest that the antibiotic activity of Jordan's red soils is due to the proliferation of antibiotic-producing bacteria.

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Farnesane-Type Sesquiterpenoids with Antibiotic Activity from Chiliadenus lopadusanus

Antibiotics ◽

10.3390/antibiotics10020148 ◽

2021 ◽

Vol 10 (2) ◽

pp. 148

Author(s):

Marco Masi ◽

Emanuela Roscetto ◽

Alessio Cimmino ◽

Maria Rosaria Catania ◽

Giuseppe Surico ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Secondary Metabolites ◽

Acinetobacter Baumannii ◽

Plant Species ◽

Antibiotic Activity ◽

Spectroscopic Methods ◽

Organic Extract ◽

Aerial Parts ◽

New Antibiotics ◽

First Time

Chiliadenus lopadusanus Brullo is an Asteraceae plant species endemic to Lampedusa island, the largest island of the Pelage archipelago, Italy. The organic extract of its whole aerial parts, showing antibiotic activity against Staphylococcus aureus and Acinetobacter baumannii, wasfractionated employing bioguided purification procedures affording three main farnesane-type sesquiterpenoids. They were identified by spectroscopic methods (NMR and ESIMS data) as the (E)-3,7,11-trimethyldodeca-1,6,10-triene-3,9-diol, (E)-10-hydroxy-2,6,10-trimethyldodeca-2,6,11- trien-4-one and (E)-10-hydroxy-2,6,10-trimethyl-dodeca-6,11-dien-4-one, commonly named 9-hydroxynerolidol, 9-oxonerolidol, and chiliadenol B, respectively. These three sesquiterpenes, isolated for the first time from C. lopadusanus, were tested on methicillin-resistant S. aureus and A. baumannii showing antibacterial and antibiofilm activities. This plant could be used as a source to isolate secondary metabolites as potential new antibiotics.

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A Cell-Free Screen for Bacterial Membrane Disruptors Identifies Mefloquine as a Novel Antibiotic Adjuvant

Antibiotics ◽

10.3390/antibiotics10030315 ◽

2021 ◽

Vol 10 (3) ◽

pp. 315

Author(s):

Jessica Podoll ◽

Justin Olson ◽

Wei Wang ◽

Xiang Wang

Keyword(s):

Staphylococcus Aureus ◽

High Throughput ◽

Antibiotic Activity ◽

Bacterial Membrane ◽

Bacterial Membranes ◽

Hit Rate ◽

New Antibiotics ◽

Lactam Antibiotic ◽

A Cell ◽

Bacterial Model

Antibacterial discovery efforts have lagged far behind the need for new antibiotics. An approach that has gained popularity recently is targeting bacterial phospholipid membranes. We leveraged the differences between bacterial and mammalian phospholipid compositions to develop a high-throughput screen that identifies agents that selectively disrupt bacterial membranes while leaving mammalian membranes intact. This approach was used to screen 4480 compounds representing a subset of the Maybridge HitFinderTM V.11 Collection and the Prestwick Chemical Drug Library®. The screen identified 35 “positives” (0.8% hit rate) that preferentially damage bacterial model membranes. Among these, an antimalarial compound, mefloquine, and an aminoglycoside, neomycin, were identified. Further investigation of mefloquine’s activity against Staphylococcus aureus showed that it has little antibiotic activity on its own but can alter membrane fluidity, thereby potentiating a β-lactam antibiotic, oxacillin, against both methicillin-susceptible and methicillin-resistant S. aureus. This study indicates that our cell-free screening approach is a promising platform for discovering bacterial membrane disruptors as antibacterials antibiotic adjuvants.

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Bioprospecting highly diverse endophytic Pestalotiopsis spp. with antibacterial properties from Maytenus ilicifolia, a medicinal plant from Brazil

Canadian Journal of Microbiology ◽

10.1139/w07-078 ◽

2007 ◽

Vol 53 (10) ◽

pp. 1123-1132 ◽

Cited By ~ 9

Author(s):

Josiane Gomes-Figueiredo ◽

Ida C. Pimentel ◽

Vânia A. Vicente ◽

Marcio R. Pie ◽

Vanessa Kava-Cordeiro ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Medicinal Plant ◽

Micrococcus Luteus ◽

Antibacterial Properties ◽

Antimicrobial Properties ◽

New Approach ◽

Pestalotiopsis Microspora ◽

Layer Chromatography ◽

Maytenus Ilicifolia ◽

Overlay Assay

Thirteen endophytic fungal strains of the genus Pestalotiopsis were isolated from the medicinal plant Maytenus ilicifolia Mart. ex. Reiss (commonly known as “espinheira santa”) and their antimicrobial properties were investigated. Two isolates were successful in inhibiting the growth of the tested microorganisms ( Escherichia coli , Klebsiella pneumoniae , Micrococcus luteus , Staphylococcus aureus , and methicillin-resistant Staphylococcus aureus (MRSA)) using the technique of bioautographic thin-layer chromatography (TLC) agar overlay assay. An analysis based on a polyphasic approach integrating taxonomic information, morphological traits, RAPD markers, and the sequencing of the ITS1–5.8S–ITS2 of the rDNA led to the assignment of the isolates as belonging to the species Pestalotiopsis microspora , Pestalotiopsis vismiae , and Pestalotiopsis leucothoes . Therefore, the present study presents a new approach to the study of endophytic fungi of the genus Pestalotiopsis.

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EFECTO INHIBITORIO DE EXTRACTOS DE Vitex mollis Kunth CONTRA BACTERIAS Y ESPECIES DE Fusarium DE IMPORTANCIA HUMANA Y AGRÍCOLA

Revista Fitotecnia Mexicana ◽

10.35196/rfm.2018.4.353-363 ◽

2018 ◽

Vol 41 (4) ◽

pp. 353-363

Author(s):

Alberto J. Valencia-Botin ◽

Melesio Gutiérrez-Lomelí ◽

Juan A. Morales-Del-Río ◽

Pedro J. Guerrero-Medina ◽

Miguel A. Robles-García ◽

...

Keyword(s):

Escherichia Coli ◽

Staphylococcus Aureus ◽

Salmonella Enterica ◽

Micrococcus Luteus ◽

E Coli

Actualmente existe la necesidad de hacer frente al problema de la resistencia a los antibióticos y al uso indiscriminado de fungicidas químicos en la agricultura. El objetivo de este trabajo fue evaluar el efecto inhibitorio de extractos acuosos, metanólicos, acetónicos y hexánicos de hoja y tallo de Vitex mollis Kunth (Lamiaceae) contra diferentes bacterias (Escherichia coli, Micrococcus luteus, Salmonella enterica y Staphylococcus aureus) y especies del hongo Fusarium (F. verticillioides, F. oxysporum, F. tapsinum y F. oxysporum f.sp. lycopersici) de importancia en la salud y en la agricultura, así como determinar su composición química general. Se determinaron las concentraciones inhibitorias mínimas (CIM) de todos los extractos por la técnica de microdilución, excepto del hexánico, que no presentó inhibición en las bacterias estudiadas. S. enterica fue la bacteria que mostró mayor sensibilidad al extracto metanólico de tallo (CIM = 28 μg mL-1), le siguieron M. luteus (CIM = 32 μg mL-1), S. aureus (CIM = 75 μg mL-1) y E. coli (CIM = 80 μg mL- 1). Los extractos metanólicos y acuosos de tallo presentaron mayor porcentaje de inhibición contra los diferentes tipos de Fusarium evaluados por el método de dilución en agar. Los extractos de V. mollis inhibieron a F. verticillioides entre 62 y 91 % con 120 μg mL-1 de extracto. El orden de las especies de hongos inhibidas por los extractos fue: F. verticillioides > F. oxysporum > F. tapsinum > F. oxysporum f.sp. lycopersici. La composición química de las especies se determinó mediante pruebas para fenoles, taninos, flavonoides, triterpenos, alcaloides, cumarinas y saponinas. Ninguno de los extractos presentó alcaloides y saponinas. Los fenoles (37.1 mg EAG/g muestra seca) y flavonoides (26.8 mg EQ/g muestra seca) fueron los compuestos mayoritarios en los extractos metanólicos y acuosos. En conclusión, se requieren cantidades muy pequeñas de extracto para la inhibición de bacterias y de Fusarium; por lo tanto, V. mollis puede ser considerada una fuente de metabolitos para este fin y en la agricultura como control alternativo dentro de un manejo integrado de enfermedades.

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An Actinobacterial Isolate, Streptomyces sp. YX44, Produces Broad-Spectrum Antibiotics That Strongly Inhibit Staphylococcus aureus

Microorganisms ◽

10.3390/microorganisms9030630 ◽

2021 ◽

Vol 9 (3) ◽

pp. 630

Author(s):

Tien-Lin Chang ◽

Tzu-Wen Huang ◽

Ying-Xuan Wang ◽

Chang-Pan Liu ◽

Ralph Kirby ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Fermentation Broth ◽

Multidrug Resistant ◽

Size Exclusion ◽

Significant Activity ◽

Health Food ◽

Strong Activity ◽

Genus Streptomyces ◽

Streptomyces Sp ◽

Good Solubility

The need for new antibiotics is increasing due to their overuse, and antibiotic resistance has become one of the major threats worldwide to public health, food safety, and clinical treatment. In this study, we describe an actinobacterial isolate, YX44, which belongs to the genus Streptomyces. This Streptomyces was isolated from a drinking pipe located in Osaka, Japan, and has the ability to inhibit Gram-positive bacteria, Gram-negative bacteria, and various fungi. YX44 fermentation broth shows strong activity against Escherichia coli and Staphylococcus aureus, as well as also inhibiting clinical isolates of multidrug-resistant Staphylococcus aureus. The YX44 antibacterial substances in the broth are relatively heat-stable, show high stability from the pH range 1 to 11, and have good solubility in both organic and non-organic solvents. Size-exclusion chromatography revealed that the YX44 antibacterial compounds are less than 1000 Da in size. LC-MS was able to identify three possible candidate molecules with molecular weights of 308, 365, 460, and 653 g/mol; none of these sizes correspond to any well-known antibiotics. Our results show that Streptomyces sp. YX44 seems to produce a number of novel antibiotics with high pH stability and good solubility that have significant activity against S. aureus, including multidrug-resistant strains.

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Multiparameter Flow Cytometric Analysis of Antibiotic Effects on Membrane Potential, Membrane Permeability, and Bacterial Counts of Staphylococcus aureus andMicrococcus luteus

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.44.4.827-834.2000 ◽

2000 ◽

Vol 44 (4) ◽

pp. 827-834 ◽

Cited By ~ 159

Author(s):

David J. Novo ◽

Nancy G. Perlmutter ◽

Richard H. Hunt ◽

Howard M. Shapiro

Keyword(s):

Staphylococcus Aureus ◽

Flow Cytometry ◽

Membrane Potential ◽

Membrane Permeability ◽

Micrococcus Luteus ◽

Flow Cytometric Analysis ◽

Bacterial Counts ◽

Flow Cytometric ◽

Permeability Parameter ◽

Particle Counts

ABSTRACT Although flow cytometry has been used to study antibiotic effects on bacterial membrane potential (MP) and membrane permeability, flow cytometric results are not always well correlated to changes in bacterial counts. Using new, precise techniques, we simultaneously measured MP, membrane permeability, and particle counts of antibiotic-treated and untreated Staphylococcus aureus andMicrococcus luteus cells. MP was calculated from the ratio of red and green fluorescence of diethyloxacarbocyanine [DiOC2(3)]. A normalized permeability parameter was calculated from the ratio of far red fluorescence of the nucleic acid dye TO-PRO-3 and green DiOC2(3) fluorescence. Bacterial counts were calculated by the addition of polystyrene beads to the sample at a known concentration. Amoxicillin increased permeability within 45 min. At concentrations of <1 μg/ml, some organisms showed increased permeability but normal MP; this population disappeared after 4 h, while bacterial counts increased. At amoxicillin concentrations above 1 μg/ml, MP decreased irreversibly and the particle counts did not increase. Tetracycline and erythromycin caused smaller, dose- and time-dependent decreases in MP. Tetracycline concentrations of <1 μg/ml did not change permeability, while a tetracycline concentration of 4 μg/ml permeabilized 50% of the bacteria; 4 μg of erythromycin per ml permeabilized 20% of the bacteria. Streptomycin decreased MP substantially, with no effect on permeability; chloramphenicol did not change either permeability or MP. Erythromycin pretreatment of bacteria prevented streptomycin and amoxicillin effects. Flow cytometry provides a sensitive means of monitoring the dynamic cellular events that occur in bacteria exposed to antibacterial agents; however, it is probably simplistic to expect that changes in a single cellular parameter will suffice to determine the sensitivities of all species to all drugs.

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PENGARUH KEMASAN TERHADAP AKTIVITAS ANTI BAKTERI SUSPENSI ERITROMISIN SETELAH PENYINARAN DENGAN SINAR MATAHARI LANGSUNG

Jurnal Ilmiah Manuntung ◽

10.51352/jim.v2i1.43 ◽

2017 ◽

Vol 2 (1) ◽

pp. 32

Author(s):

Nurul Hidayati

Keyword(s):

Staphylococcus Aureus ◽

Dosage Form ◽

Antibiotic Activity ◽

Test Solution ◽

Diffusion Method ◽

Direct Sunlight ◽

Closed Container ◽

Microbiological Test ◽

Hydrolysis Process ◽

The Stability

Erythromycin is a type of antibiotic medication. The drug is unstable when in the water because it can undergo hydrolysis process. Fortunately, it can be prevented by making dry syrup dosage form. In addition to hydrolysis, erythromycin can also have photolysis. For preventing the reaction, it can be done by packing the medication in an opaque and tightly closed container. This research was conducted to determine the effect of packaging on the stability of erythromycin suspension that has been exposed to direct sunlight within a period. The research was done by exposing erythromycin suspension, which was packed with three kinds of color packaging namely brown, translucent, and opaque, for 30 minutes, 60 minutes, 90 minutes and 120 minutes, then continued by a microbiological test with a diffusion method to determine the activity of erythromycin against Staphylococcus Aureus. The results of the research showed the existence of diameter of barrier areas around the spots that were given a test solution. It means that the test solution has antibiotic activity against bacteria. Preparations packaged in brown color have higher inhibition when compared with the opaque and translucent. It shows that the erythromycin suspension packaged in brown colors was more efficient in stabilizing the preparation

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Future directions for the discovery of antibiotics from actinomycete bacteria

Emerging Topics in Life Sciences ◽

10.1042/etls20160014 ◽

2017 ◽

Vol 1 (1) ◽

pp. 1-12 ◽

Cited By ~ 8

Author(s):

Rebecca Devine ◽

Matthew I. Hutchings ◽

Neil A. Holmes

Keyword(s):

New Technologies ◽

Gene Clusters ◽

Biosynthetic Gene Clusters ◽

Future Directions ◽

Streptomyces Species ◽

Genus Streptomyces ◽

New Antibiotics ◽

The Uk ◽

Almost All ◽

New Generation

Antimicrobial resistance (AMR) is a growing societal problem, and without new anti-infective drugs, the UK government-commissioned O'Neil report has predicted that infectious disease will claim the lives of an additional 10 million people a year worldwide by 2050. Almost all the antibiotics currently in clinical use are derived from the secondary metabolites of a group of filamentous soil bacteria called actinomycetes, most notably in the genus Streptomyces. Unfortunately, the discovery of these strains and their natural products (NPs) peaked in the 1950s and was then largely abandoned, partly due to the repeated rediscovery of known strains and compounds. Attention turned instead to rational target-based drug design, but this was largely unsuccessful and few new antibiotics have made it to clinic in the last 60 years. In the early 2000s, however, genome sequencing of the first Streptomyces species reinvigorated interest in NP discovery because it revealed the presence of numerous cryptic NP biosynthetic gene clusters that are not expressed in the laboratory. Here, we describe how the use of new technologies, including improved culture-dependent and -independent techniques, combined with searching underexplored environments, promises to identify a new generation of NP antibiotics from actinomycete bacteria.

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