scholarly journals Isolation, screening and identification of microorganisms having antimicrobial activity isolated from samples collected on seabed of Northeast Vietnam

2016 ◽  
Vol 14 (3) ◽  
pp. 539-547
Author(s):  
Lê Thị Hồng Minh ◽  
Vũ Thị Quyên ◽  
Nguyễn Mai Anh ◽  
Đoàn Thị Mai Hương ◽  
Brian T Murphy ◽  
...  

Microorganisms are especially interested in due to the ability to produce secondary compounds with high-value applications. Plenty of novel and diverse chemical structures have been found in the bioactive substances of microorganisms. In this study, we isolated 143 strains of bacteria and actinomycetes from 161 samples including: sediments, sponges, soft corals, echinoderms and starfish collected from three sea areas of Viet Nam: Ha Long - Cat Ba; Co To - ThanhLan; Bai Tu Long. The strains were fermented in A1 medium and then fermentation broths were extracted 5 times with ethyl acetate. The extraction residue screening test using 7 reference strains isolated 15 target strains with the highest biological activity. Most of these strains have dramatic inhibition on Gram positive bacteria: Enterococcus faecalis ATCC29212; Bacillus cereus ATCC13245  and Candida albicans ATCC10231 with MIC values  ​​less than or equal to the MIC value of the reference antibiotic. In particular, strain G057 was active against S. enterica ATCC 13076 and G002 inhibited E. coli ATCC25922 with respective values  ​​MICG057 = 8 µg/ ml, MICG002 = 256µg/ ml; and three strains G115, G119, G120 showed the inhibitory effect towards P. aeruginosa ATCC27853 with respective values ​​MICG115 = 64 µg/ ml, MICG119 = 32 µg/ ml and MICG120 = 32 µg/ ml. All 15 strains were then subjected to morphological and phylogenetic investigations based on 16S rRNA gene sequences. The results showed that 9 of 15 strains G016, G017, G019, G043, G044, G047, G068, G119 and G120 belonged to Genus Micromonospora; strains G039 and G065 were identified as Genus Stretomyces; G002 was  identified as Bacillus; G057 was  identified as Nocardiopsis; G115 was in Photobacterium and G121 belonged Oceanisphaera.

2018 ◽  
Vol 15 (4) ◽  
pp. 737-744
Author(s):  
Cao Duc Tuan ◽  
Le Thi Hong Minh ◽  
Vu Thi Quyen ◽  
Nguyen Mai Anh ◽  
Doan Thi Mai Huong ◽  
...  

Microorganisms are of particular interest because of their ability to synthesize high-value secondary compounds and provide us with novel and diverse chemical structures. The most common source of antibiotics is Actinomycetes which provide around two-third of naturally occurring antibiotics, including many of medical importance. In this study, 81 strains of actinomycetes were isolated from 145 samples including: sediments, sponges, soft corals, echinoderms and starfish collected from three sea areas of Vietnam: Hue, Da Nang and Quang Nam. The strains were fermented in A+ medium and fermentation broths were extracted 5 times with ethyl acetate. The extracts were evaporated under reduced  pressure to yield crude extracts. Quantitative assay was used to determine MIC (Minimum inhibitory concentration) of extract against 7 reference strains. From the results of screening, Seven strains of actinomycetes that have the highest biological activity (Code: G244, G246, G261, G266, G278, G280 and G290) were chosen to be identified by morphological and phylogenetic based on 16S rRNA gene sequences. The results showed that 6 strains G246, G261, G266, G278, G280 and G290 belonged to the genus Streptomyces; and the strain G244 belonged to the genus Micromonospora. In particular, strains G244, G278, G280 were resistant 5/7 strains of microorganisms test, with values  MICs from 2 µg/mL to 256 µg/mL; and three strains G261, G266, G290 showed the inhibitory effect towards 4/7 strains of microorganisms test, with respective values MICs from 2 µg/mL to 256 µg/mL. Moreover, six of the seven selected strains were highly resistant to yeast Candida albicans ATCC10231 with MIC values from 2 µg/mL to 256 µg/mL. These results indicated that marine Actinomycetes in Vietnam are also a potential source to find bioactive substances.


2021 ◽  
Vol 19 (3) ◽  
pp. 557-568
Author(s):  
Vu Thi Quyen ◽  
Vu Thi Thu Huyen ◽  
Nguyen Mai Anh ◽  
Nguyen Hai Dang ◽  
Doan Thi Mai Huong ◽  
...  

Actinomycetes has been extensively studied due to its ability to produce secondary compounds with high application value. Especially their antibiotic ability, more than 40% of antibiotics are derived from actinomycetes of which genus Streptomyces predominates. Moreover, marine actinomycetes have been being of great interest in recent years due to their ability to produce bioactive substances capable of providing diverse and novel chemical structures. In this study, from 8 samples of marine sediments collected in Vung Ang bay in Ha Tinh provinces, we have isolated 20 strains of actinomycetes. The strains were fermented in A1+ medium, the fermentation fluid was extracted 5 times with ethyl acetate, recovered sediment and determined antibacterial and cytotoxic activity. From the screening results, two strains with the highest antibacterial activity and highest cytotoxicity were selected ie., HT03 and HT06. Both strains had antagonistic activity of Enterococcus faecalis ATCC29212 with MICHT03= 32 μg/mL, MICHT06= 16μg/mL, with Stapphylococus aureus ATCC25923 with MICHT03= 64 μg/mL, MICHT06= 32 μg/mL and with Bacillus cereus ATCC 13245 has the same MIC = 16 μg / mL. In addition, the two strains HT03 and HT06 were able to strongly inhibit the yeast Candida albicans ATCC10231 with MICHT03= 16 μg/mL, MICHT06= 8 μg/mL. Especially, two strains, HT03 and HT06, exhibited very good toxicity on all 5 cancer cell lines (MCF-7 breast cancer cell; MDA-MB-231 breast cancer cell; lung cancer cell). NCI-H1975; HeLa cervical cancer cell; AGS gastric cancer cell) at both test concentrations of 30 µg/mL and 100µg/mL. By the analysis of 16S rRNA sequence, the results showed that the HT03 strain had the highest similarity (99.93%) to that of Streptomyces fradiaes and Streptomyces fradiae ATCC. The HT06 strain was defined to belong to Nocardiopsis synnemataformans with the 16S sequence identity of 99.89% to the Japanese standard Nocardiopsis synnemataformans DSM 44143 strain NBRC-102581.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Pedro Seguí ◽  
John J. Aguilera-Correa ◽  
Elena Domínguez-Jurado ◽  
Christian M. Sánchez-López ◽  
Ramón Pérez-Tanoira ◽  
...  

AbstractThis study was designed to propose alternative therapeutic compounds to fight against bacterial pathogens. Thus, a library of nitrogen-based compounds bis(triazolyl)methane (1T–7T) and bis(pyrazolyl)methane (1P–11P) was synthesised following previously reported methodologies and their antibacterial activity was tested using the collection strains of Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, and Pseudomonas aeruginosa. Moreover, the novel compound 2P was fully characterized by IR, UV–Vis and NMR spectroscopy. To evaluate antibacterial activity, minimal inhibitory concentrations (MICs), minimal bactericidal concentrations (MBCs), minimum biofilm inhibitory concentrations (MBICs), and minimum biofilm eradication concentrations (MBECs) assays were carried out at different concentrations (2–2000 µg/mL). The MTT assay and Resazurin viability assays were performed in both human liver carcinoma HepG2 and human colorectal adenocarcinoma Caco-2 cell lines at 48 h. Of all the synthesised compounds, 2P had an inhibitory effect on Gram-positive strains, especially against S. aureus. The MIC and MBC of 2P were 62.5 and 2000 µg/mL against S. aureus, and 250 and 2000 µg/mL against E. faecalis, respectively. However, these values were > 2000 µg/mL against E. coli and P. aeruginosa. In addition, the MBICs and MBECs of 2P against S. aureus were 125 and > 2000 µg/mL, respectively, whereas these values were > 2000 µg/mL against E. faecalis, E. coli, and P. aeruginosa. On the other hand, concentrations up to 250 µg/mL of 2P were non-toxic doses for eukaryotic cell cultures. Thus, according to the obtained results, the 2P nitrogen-based compound showed a promising anti-Gram-positive effect (especially against S. aureus) both on planktonic state and biofilm, at non-toxic concentrations.


2021 ◽  
Vol 22 (10) ◽  
Author(s):  
Alvita Indraswari ◽  
I Wayan Suardana ◽  
Aris Haryanto ◽  
Dyah Ayu Widiasih

Abstract. Indraswari A, Suardana IW, Haryanto A, Widiasih DA. 2021. Molecular analysis of pathogenic Escherichia coli isolated from cow meat in Yogyakarta, Indonesia using 16S rRNA gene. Biodiversitas 22: 4566-4573. Meat has been recognized as a major source of foodborne disease and a public health problem. The characteristics of meat become an ideal growth medium for various microorganisms if not handled properly. Pathogenic Escherichia coli is one of the foodborne disease agents that causes diarrhea. Identification of pathogenic E. coli isolated from cow meat needs to be done. This research aims to study nucleotide sequence of 16S rRNA gene of pathogenic E. coli isolated from cow meat in Yogyakarta, Indonesia using Polymerase Chain Reaction (PCR). These fifteen isolates have been detected for eae target gene, then amplification of the 16S rRNA gene was carried out using primers 27F and 1492R. Phylogenetic tree reconstruction was performed on the fifteen isolates of pathogenic E. coli to figure out the relationship to reference strains available at the GenBank. Results show that nucleotide sequence among the fifteen isolates from different traditional markets in Yogyakarta, Indonesia and reference strains are very similar. The fifteen isolates have small genetic distance to the reference strains, and these fifteen isolates are also in the same clade with reference strains. This research shows that the fifteen isolates under investigation are closely related to the reference strains, which is Shiga-toxin producing E. coli (STEC). People should pay more attention in processing food stock, especially cow meat. Further research may focus on determining the strain of those fifteen isolates.


1989 ◽  
Vol 52 (9) ◽  
pp. 665-667 ◽  
Author(s):  
R. S. FARAG ◽  
Z. Y. DAW ◽  
F. M. HEWEDI ◽  
G. S. A. EL-BAROTY

Six spice essential oils (sage, rosemary, caraway, cumin, clove, and thyme) and their basic ingredients were tested for their inhibitory effect against 3 strains of Gram-negative bacteria, 4 strains of Gram-positive bacteria, one acid fast bacterium, and one yeast. Preliminary screening of antimicrobial activity of the essential oils was done using the filter paper disc agar diffusion method. The minimum inhibitory concentration for each essential oil against various micro-organisms was also measured. Very low concentrations (0.25 – 12 mg/ml) of the various essential oils were sufficient to prevent microbial growth. The data show that Gram-positive bacteria were more sensitive to the antimicrobial compounds in spices than Gram-negative. The inhibition zones of different microbial growth produced by various essential oils were similar to those produced by their basic compounds. Thyme and cumin oils possessed very strong antimicrobial activity compared with the other essential oils. There was a relationship between the chemical structures of the most abundant compounds in the essential oils under investigation and the antimicrobial activity.


2018 ◽  
Vol 55 (5A) ◽  
pp. 66
Author(s):  
Hai Van Nguyen

In this study, the interaction effects of Litsea cubeba essential oil (EO) and two antibiotics (ABs) on the antibacterial activity against pathogenic bacteria in aquaculture were investigated. Two reference strains (Escherichia coli ATCC 25922 (E. coli ATCC 25922) and Vibrio parahaemolyticus ATCC 17802 (V. parahaemolyticus ATCC 17802)) and six locally isolated aquatic pathogenic strains (E. coli 9C48, E. coli 11C123, Vibrio 2S4, Vibrio 2N38, V. parahaemolyticus ND201 and V. parahaemolyticus TB81) were used. The inhibitory effects of individual antimicrobial agent (L. cubeba EO, nalidixic acid, and oxytetracycline) were tested against eight strains by using broth microdilution assay in 96-well microplates. A higher inhibitory effect of L. cubeba EO was observed against isolated aquatic pathogenic (MIC = 1.15 - 2.30 mg/mL) than that in the reference strains (MIC = 5.53 mg/mL). The combination effects of L. cubeba EO and ABs often used in the treatment of bacteria effects in aquaculture (nalidixic acid and oxytetracycline) were evaluated by the checkerboard method. Fractional Inhibitory Concentration (FIC) values were determined to characterize the interaction among combinations. Out of 16 AB-EO combinations tested, 11 of them showed a synergistic effect (FIC ≤ 0.5), 3 of them showed an additional effect (0.5 < FIC ≤ 1.0) and 2 of them showed indifferent effect           (1 < FIC ≤ 4), no antagonistic effect was observed. The antimicrobial synergy of EO and AB could enhance efficacy, reduce toxicity, decrease adverse side effects, and lower the dose of ABs used in aquaculture.


2020 ◽  
Vol 16 (4) ◽  
pp. 721-728
Author(s):  
Le Thi Hong Minh ◽  
Nguyen Mai Anh ◽  
Vu Thi Quyen ◽  
Vu Thi Thu Huyen ◽  
Doan Thi Mai Huong ◽  
...  

Marine environment is rich in natural product resources, including marine microorganisms, especially fungi which are not only seen as a potential source of highly applicable bioactive substances but also can provide for science new chemical structures. The objective of this study is to isolate and screen fungal strains with antibacterial activity from the marine environment. Twenty five strains of fungi were isolated from marine sediments of Thanh Lan, Co To island and assessed on antibiotic activity against 7 tested microbial strains, including three Gram-negative bacteria (Escherichia coli ATCC25922, Pseudomonas aeruginosa ATCC27853, Salmonella enterica ATCC13076), three Gram-positive bacteria (Enterococcus faecalis ATCC29212, Stapphylococus aureus ATCC25923, Bacillus cereus ATCC 13245), and the yeast Candida albicans ATCC10231. The minimum inhibitory concentration (MIC) against the tested microorganisms was determined for the crude extracts obtained from the culture broths after ethyl acetate extraction and vacuum rotary evaporation. Three strains with the highest antimicrobial activity M26, M30 and M45 were capable of inhibiting 4 - 5 of the 7 tested microorganisms with MIC values from 64 to 256 μg/ml, depending on each tested strain. Morphological and phylogenetic investigations based on 18S rRNA gene sequences of the three selected strains showed that strains M26 and M30 belonged to the genus Penicillium, whereas strain M45 belonged to the genus Neurospora. The sequences of 18S rRNA gene of three strains M26, M30 and M45 were registered on GenBank database with accession numbers: MH673730, MH673731, MH673732, respectively. Research results showed that marine environment has a great potential in isolation of fungal strains for the search for antibacterial substances as well as other biologically active compounds.


2010 ◽  
Vol 4 (1) ◽  
pp. 140-146 ◽  
Author(s):  
Germano Orrù ◽  
Susanna Del Nero ◽  
Enrica Tuveri ◽  
Maria Laura Ciusa ◽  
Francesca Pilia ◽  
...  

A dental unit water line (DUWL) equipped with a device designed to automatically and continually flush a bacteriostatic solution of hydrogen peroxide (WHE) and a discontinuous disinfecting system (BIOSTER) was evaluated. In the first instance a preliminary sensitivity test on a large number of microorganisms (bacteria and fungi) was tried with a H2O2 range from 100 to 800 ppm. The bacteria frequently reported in DUWL (including Pseudomonas spp, Streptococcus spp., Staphylococcus spp., E. coli) and some periodontal pathogens showed a minimum inhibitory concentration from 100 to 300 H2O2 ppm (also including M. marinum and C. albicans). However, H2O2 did not show any inhibitory effects against: A. actinomycetemcomitans, C. glabrata C. parapsilos, F. nucleatum, M. micros. In a second step, the DUWL was experimentally infected with S. faecalis, E. coli, P. aeruginosa, S. aureus. After disinfection steps with 3% H2O2, the inhibitory effect on planktonic forms and on sessile biofilm was measured. In a third step, the count of 16S rRNA gene copies by real time PCR at different points of the DUWL described an accrue of bacterial slime in “hot spot” regions characterized by irregular/slow water flux (valves, elbows). However these results suggest that hydrogen peroxide is not only able to inhibit bursts of planktonic bacteria inside the DUWL, but that it could also be effective against sessile biofilm containing heterotrophic microorganisms derived from domestic water line contamination. In addition some oral pathogens could be contaminating and surviving in DUWL.


2006 ◽  
Vol 72 (3) ◽  
pp. 1932-1938 ◽  
Author(s):  
Ola A. Olapade ◽  
Morgan M. Depas ◽  
Erika T. Jensen ◽  
Sandra L. McLellan

ABSTRACT A high biomasses of Cladophora, a filamentous green alga, is found mainly during the summer along the shores of Lake Michigan. In this study, the abundance and persistence of the fecal indicator bacterium Escherichia coli and sulfate-reducing bacteria (SRB) on Cladophora mats collected at Lake Michigan beaches were evaluated using both culture-based and molecular analyses. Additionally, 16S rRNA gene cloning and sequencing were used to examine the bacterial community composition. Overall, E. coli was detected in all 63 samples obtained from 11 sites, and the average levels at most beaches ranged from 2,700 CFU/100 g (wet weight) of Cladophora to 7,500 CFU/100 g of Cladophora. However, three beaches were found to have site average E. coli densities of 12,800, 21,130, and 27,950 CFU/100 g of Cladophora. The E. coli levels in the lake water collected at the same time from these three sites were less than the recommended U.S. Environmental Protection Agency limit, 235 CFU/100 ml. E. coli also persisted on Cladophora mats in microcosms at room temperature for more than 7 days, and in some experiments it persisted for as long as 28 days. The SRB densities on Cladophora mats were relatively high, ranging from 4.4 × 106 cells/g (6.64 log CFU/g) to 5.73 × 106 cells/g (6.76 log CFU/g) and accounting for between 20% and 27% of the total bacterial counts. Partial sequences of the 16S rRNA gene clones revealed a phylogenetically diverse community, in which the Cytophaga-Flavobacterium-Bacteroides cluster and the low-G+C-content gram-positive bacteria were the dominant organisms, accounting for 40% and 12.8%, respectively, of the total clone library. These results further reveal the potential public health and ecological significance of Cladophora mats that are commonly found along the shoreline of Lake Michigan, especially with regard to the potential to harbor microorganisms associated with fecal pollution and odor-causing bacteria.


Foods ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2367
Author(s):  
Yanpeng Hao ◽  
Zuchen Wei ◽  
Zhi Wang ◽  
Guiying Li ◽  
Yang Yao ◽  
...  

Coarse cereals are rich in flavonoids, which are bioactive substances with a wide range of functions. Biotransformation is considered an emerging approach to methylate flavonoids, displaying prominent regio- and stereoselectivity. In the current study, liquiritigenin, naringenin, and hesperidin flavonoids were biotransformed using O-methyltransferases that were heterologously expressed in Saccharomyces cerevisiae BJ5464-NpgA. Nuclear magnetic resonance (NMR) spectroscopy was used together with high-resolution mass spectroscopy analysis to determine the structures of the resulting methylated transformants, and their antimicrobial and antiproliferation activities were also characterized. Among the five methylated flavonoids obtained, 7-methoxy-liquiritigenin had the strongest inhibitory effect on Candida albicans SC5314 (C. albicans SC5314), Staphylococcus aureus ATCC6538 (S. aureus ATCC6538), and Escherichia coli ATCC25922 (E. coli ATCC25922), which increased 7.65-, 1.49-, and 0.54-fold in comparison to the values of their unmethylated counterparts at 200, 250, and 400 μM, respectively. The results suggest that 3’-methoxyhesperetin showed the best antiproliferative activity against MCF-7 cells with IC50 values of 10.45 ± 0.45 µM, which was an increase of more than 14.35-fold compared to that of hesperetin. These results indicate that methylation enhances the antimicrobial activities and antiproliferative effects of flavonoids. The current study provides an experimental basis for further research on flavonoids as well as flavonoid-containing crops in the development of antimicrobial and anti-breast cancer drugs in addition to supplementary and health foods. The biotransformation method is ideal, as it represents a means for the sustainable production of bioactive flavonoids.


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