scholarly journals Isolation, characterization, and molecular identification of soil bacteria showing antibacterial activity against human pathogenic bacteria

Author(s):  
R. Prashanthi ◽  
Shreevatsa G.K. ◽  
Krupalini S. ◽  
Manoj L.

Abstract Background The present study dealt with the screening of soil bacteria with antibacterial activity from different locations in Bangalore, India. Antibiotics play the role of self-defense mechanism for the bacteria and are produced as secondary metabolites to protect themselves from other competitive microorganisms. The need for new antibiotics arose as the pathogenic bacteria acquire resistance to various antibiotics meant for treating human diseases. Given the importance of antibiotics of bacterial origin, standard techniques have been used to isolate and characterize the soil bacteria which showed antibacterial activity. Results The isolated bacteria were tested against human pathogenic bacteria like Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae by primary and secondary screening methods. The isolates PR1, PR2, and PR3 were confirmed to have antibacterial activity against S. aureus, E. coli, P. aeruginosa, and K. pneumoniae by both methods. Studies on the effect of filter sterilization, autoclaving, and proteinase K treatment on culture filtrates showed filter sterilization as the best method. The effect of different carbon and nitrogen sources on the antibacterial activity showed that preference by each isolate differed for carbon and nitrogen requirements. The isolates PR1, PR2, and PR3 were identified as Bacillus aryabhattai strain PR-D07, Arthrobacter humicola strain PR-F07, and Neomicrococcus lactis strain PR-F11 through 16S rRNA sequencing. Conclusion Findings from this research work are encouraging and could proceed further to applied aspects. Only 3 bacterial isolates out of 263 isolates from soil samples displayed antibacterial activity against human pathogens S. aureus, E. coli, P. aeruginosa, and K. pneumoniae. They were identified as B. aryabhattai, A. humicola, and N. lactis by 16S rRNA studies and all of them are Gram-positive. Each isolate preferred different carbon and nitrogen sources for their enhanced antibacterial activity. Efficacy of the culture filtrates of these isolates was tested by filter sterilization, autoclaving, and proteinase K treatment. Filter-sterilized culture filtrates showed higher antibacterial activity than other treatments. A comparison of the antibacterial activity of culture filtrates and antibiotic streptomycin produced an inhibition zone of 18.5 mm and 15.5 mm respectively. This is the first report on the antibacterial activity of all the 3 bacterial strains (B. aryabhattai strain PR-D07, A. humicola strain PR-F07, and N. lactis strain PR-F11), against all the human pathogens, mentioned earlier. It is also found that the antibiotic factor is proteinaceous as proteinase K considerably reduced the antibacterial activity of the culture filtrates. With the above significant results, these 3 bacteria are considered to be promising candidates for the isolation of new antibacterial agents.

2015 ◽  
Vol 18 (1) ◽  
pp. 61-65 ◽  
Author(s):  
Md Uzzal Haque ◽  
Md Ajijur Rahman ◽  
Md Anwarul Haque ◽  
Ashish Kumar Sarker ◽  
Md Anwar Ul Islam

In this study, we investigated the effect of pathogenic bacteria on the production of antibiotics by actinomycetes. We used four strains of actinomycetes-ANAM-5, ANAM-39, AIAH-10 and ANTS-1, which were isolated from the soils of Sundrabans, Bangladesh. All the strains were cultured in absence or presence of either alive or heatkilled human pathogens (Staphylococcus aureus and E. coli). The antibacterial activity of cultured cell-free supernatant fluid was analyzed by disc diffusion assay against the inducer strains. Three out of the four marine actinomycetes tested showed enhanced antibacterial activity against Staphylococcus aureus and E. coli. ANAM-5 and AIAH-10 showed enhanced antibacterial activity when co-cultured with Staphylococcus aureus whereas ANAM-5 and ANAM-39 showed enhanced antibacterial activity when co-cultured with E. coli. The highest enhancement of antibacterial activity was exhibited by the strain ANAM-5 against Staphylococcus aureus (from 9 mm to 18 mm inhibition zone). The study has important ecological and biotechnological implications in case of microbial competition in the natural environment and may become helpful to discover novel classes of antibiotics with high specificity and huge production.Bangladesh Pharmaceutical Journal 18(1): 61-65, 2015


2018 ◽  
Vol 1 (1) ◽  
pp. a40-44
Author(s):  
JIAZHEN LIM ◽  
YANG LEE ◽  
BADIOZAMAN SULAIMAN ◽  
LESLEY MAURICE BILUNG ◽  
YEE LING CHONG

The epidermal mucus of fish contains antimicrobial agents that act as biological defence against disease. This study aims to identify antibacterial activity and protein concentration of epidermal mucus of Barbodes everetti, a Bornean endemic freshwater fish. The epidermal mucus was extracted with 3% acetic acid, 0.85% sodium chloride and crude solvents. The mucus activity against eight strains of human pathogenic bacteria, including Bacillus cereus ATCC 33019, Escherichia coli O157:H7, Listeria monocytogenes ATCC 7644, Pseudomonas aeruginosa ATCC 27853, Salmonella braenderup ATCC BAA 664, Salmonella typhimurium, Staphylococcus aureus ATCC 25933, and Vibrio cholerae, were tested. The acetic acid mucus extract of B. everetti was able to inhibit five strains of bacteria and show no activity toward E. coli O157:H7, B. cereus ATCC 33019 and L. monocytogenes ATCC 7644. Moreover, the highest protein concentration was quantified in crude extract, followed by aqueous and acetic acid extracts. This study provides a preliminary knowledge on the activity of epidermal mucus of B. everetti towards five out of the eight human pathogens tested, therefore it may contain potential sources of novel antibacterial components which could be further extracted for the production of natural antibiotics towards human-related pathogenic bacteria. Keywords: Antibacterial, antimicrobial properties, Barbodes everetti, epidermal mucus, pathogenic bacteria


Author(s):  
Pramod Dhakal ◽  
Ankit a Achary ◽  
Vedamurthy Joshi

Bioenhancers are drug facilitator which do not show the typical drug activity but in combination to enhance the activity of other molecule in several way including increase the bioavailability of drug across the membrane, potentiating the drug molecules by conformational interaction, acting as receptor for drug molecules and making target cell more receptive to drugs and promote and increase the bioactivity or bioavailability or the uptake of drugs in combination therapy. The objective of the present study was to evaluate the antibacterial and activity of combination in Azadirachta indica extract with cow urine distillate and pepper extract against common pathogenic bacteria, a causative agent of watery diarrhea. It has been found that Indian indigenous cow urine and its distillate also possess bioenhancing ability. Bioenhancing role of cow urine distillate (CUD) and pepper extract was investigated on antibacterial activity of ethanol extract of Azadirachta indica. Antibacterial activity of ethanol extract neem alone and in combination with CUD and pepper extract were determined the ATCC strains against Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa and E-coli by cup plate diffusion method. Ethanol extract of neem has showed more effect on P. aeruginosa, E-coli than S. aureus and K. pneumonia with combination of CUD and pepper extract. CUD and pepper did not show any inhibition of test bacteria in low concentration. The antibacterial effect of combination of extract and CUD was higher than the inhibition caused by extract alone and is suggestive of the bioenhancing role of cow urine distillate and pepper. Moreover, inhibition of test bacteria was observed with less concentration of extract on combining with CUD


Author(s):  
ANNAMALAI MADURAM ◽  
RAJU KAMARAJ

Objectives: The objectives of the study were to study the antibacterial activity for the various extracts of Clausena dentata against human pathogens. Clausena (Rutaceae) is a genus of about 23 species of unarmed trees and shrubs. The stem bark of C. dentata is used in veterinary medicine for the treatment of wounds and sprains. Even though C. dentata has a lot of potential medical uses, the study of microbiological properties is very scarce. Methods: The plant C. dentata was collected from Kadagaman, near Tiruvannamalai, Tamil Nadu, India, and authenticated by Centre for Advanced Study in Botany, University of Madras, Chennai. The dry powder of stem bark was extracted with hexane, chloroform, and methanol. The extracts were subjected to qualitative phytochemical screening and antibacterial activity against human pathogenic bacteria such as Escherichia coli, Salmonella Typhi, Klebsiella pneumonia, Vibrio cholerae, and Staphylococcus aureus and compared with ciprofloxacin. Results: Qualitative chemical tests revealed the presence of various phytochemicals such as alkaloids, glycosides, carbohydrate, proteins and amino acids, phytosterols, and volatile oil. The antibacterial activity result reveals that all the extracts were are more active against V. cholerae. The activity against Pseudomonas aeruginosa was mild. Conclusion: The activity against V. cholerae was comparable with that of 5 μg/mL ciprofloxacin at the concentration of C. dentata 40 μg/mL. The orders of antibacterial activity against human pathogenic bacteria are hexane, methanol, and chloroform extract of C. dentata.


Antibiotics ◽  
2018 ◽  
Vol 7 (4) ◽  
pp. 98 ◽  
Author(s):  
Eunice Mgbeahuruike ◽  
Pia Fyhrquist ◽  
Heikki Vuorela ◽  
Riitta Julkunen-Tiitto ◽  
Yvonne Holm

Piper guineense is a food and medicinal plant commonly used to treat infectious diseases in West-African traditional medicine. In a bid to identify new antibacterial compounds due to bacterial resistance to antibiotics, twelve extracts of P. guineense fruits and leaves, obtained by sequential extraction, as well as the piperine and piperlongumine commercial compounds were evaluated for antibacterial activity against human pathogenic bacteria. HPLC-DAD and UHPLC/Q-TOF MS analysis were conducted to characterize and identify the compounds present in the extracts with promising antibacterial activity. The extracts, with the exception of the hot water decoctions and macerations, contained piperamide alkaloids as their main constituents. Piperine, dihydropiperine, piperylin, dihydropiperylin or piperlonguminine, dihydropiperlonguminine, wisanine, dihydrowisanine and derivatives of piperine and piperidine were identified in a hexane extract of the leaf. In addition, some new piperamide alkaloids were identified, such as a piperine and a piperidine alkaloid derivative and two unknown piperamide alkaloids. To the best of our knowledge, there are no piperamides reported in the literature with similar UVλ absorption maxima and masses. A piperamide alkaloid-rich hexane leaf extract recorded the lowest MIC of 19 µg/mL against Sarcina sp. and gave promising growth inhibitory effects against S. aureus and E. aerogenes as well, inhibiting the growth of both bacteria with a MIC of 78 µg/mL. Moreover, this is the first report of the antibacterial activity of P. guineense extracts against Sarcina sp. and E. aerogenes. Marked growth inhibition was also obtained for chloroform extracts of the leaves and fruits against P. aeruginosa with a MIC value of 78 µg/mL. Piperine and piperlongumine were active against E. aerogenes, S. aureus, E. coli, S. enterica, P. mirabilis and B. cereus with MIC values ranging from 39–1250 µg/mL. Notably, the water extracts, which were almost devoid of piperamide alkaloids, were not active against the bacterial strains. Our results demonstrate that P. guineense contains antibacterial alkaloids that could be relevant for the discovery of new natural antibiotics.


Author(s):  
Selvarani Murugan

Objective: Resistance to antibacterial agents by pathogenic bacteria has emerged in recent years and is a major challenge for the healthcare industry. Copper nanoparticles (CuNPs) are known to be one of the multifunctional inorganic nanoparticles with effective antibacterial activity. Hence the present investigation has been focused on synthesizing and evaluating the bactericidal effect of copper nanoparticles.Methods: CuNPs were synthesized by reducing the aqueous solution of copper sulfate with sodium borohydride. The synthesized particles were characterized by x-ray diffractogram (XRD), scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) techniques to analyze size, morphology and quantitative information respectively. The antibacterial activity of CuNPs was examined by agar well diffusion method. Synergistic effect of CuNPs with broad-spectrum antibiotics was determined by the agar disc diffusion method.Results: Color change of reaction mixture from blue to dark brown indicated the formation of CuNPs. SEM image clearly demonstrated that the synthesized particles were spherical in shape and its size was found to be 17.85 nm. EDS report confirmed the presence of elemental copper in the resultant nanoparticles and its accounts for major proportion (96%) of the mass of nanoparticles. Bacterial effect of CuNPs revealed that Pseudomonas aeruginosa showed the highest antibacterial sensitivity (16.00±1.63 mm), whereas least susceptibility (9.67±0.47 mm) was noticed against Staphylococcus aureus. An enhanced antibacterial activity of commercial antibiotics was also noticed when it combined with CuNPS. A minimum zone of inhibition was increased from 0.67±0.47 mm to 10.66±0.24 mm when the nanoparticles and antibiotics were given together.Conclusion: It was observed that copper nanoparticles exhibited profound activity against all the tested bacterial strains which shows that CuNPs may serve as a better option for use in medicine in the future.


2016 ◽  
Author(s):  
Wenfa Ng ◽  
Yen-Peng Ting

Microbes for environmental research should be cultured in growth media with characteristics (e.g., pH, ionic strength, and organic and ionic composition) as close to their original habitat as possible. Additionally, the medium should also enable high cell density to be obtained - needed for providing sufficient cells in subsequent experiments. This in-progress report describes the formulation of a medium with an environmentally-relevant composition (lack of complex organics), and that allows aerobic high cell density cultivation of Escherichia coli DH5α in shake flasks. The formulated medium comprises four components: a buffer system (K2HPO4: 12.54 g/L and KH2PO4: 2.31 g/L), vitamins (yeast extract: 12.0 g/L), salts (NaCl: 5.0 g/L and MgSO4: 0.24 g/L), and carbon and nitrogen sources (D-Glucose: 6.0 g/L and NH4Cl: 1.5 g/L). Notable characteristics of this medium were: high capacity phosphate buffer system (89 mM phosphate); 1:1 molar ratio between D-Glucose and NH4Cl; and yeast extract providing trace elements and a secondary carbon and nitrogen source. Growth experiments revealed that an OD600nm of 9 was attained after 24 hours of cultivation at 37 oC. Glucose and NH4Cl serve as primary carbon and nitrogen sources for this phase of growth. After 48 hours, the OD600nm reached 11, where carbohydrates, lipids and proteins in yeast extract provided the nutrients for biomass formation. Broth’s pH varied between 5.5 and 7.8 during cultivation, which was in the range conducive for E. coli growth. In comparison, the OD600nm of E. coli reached 1.4, 3.2, and 9.2 in three commonly used complex media; Nutrient Broth, LB Lennox, and Tryptic Soy Broth, respectively, over 48 hours under identical culture conditions. In addition, the formulated medium was able to maintain a large viable cell population for a longer period of time (three days) compared to Tryptic Soy Broth. Thus, preliminary data suggested that the formulated medium holds potential for use as a high cell density aerobic growth medium for Gram-negative bacteria.


2014 ◽  
Vol 77 (9) ◽  
pp. 1599-1604 ◽  
Author(s):  
MAHBOUBEH MIRHOSSEINI ◽  
VAHID ARJMAND

Practical applications of different concentrations (0, 1, 2, 4, 6, and 8 mM) of zinc oxide (ZnO) suspensions containing 1% acetic acid were investigated against the pathogenic bacteria Listeria monocytogenes, Escherichia coli, Staphylococcus aureus, and Bacillus cereus. ZnO suspensions (0, 1, 3, 6, and 8 mM) containing acetic acid had a significant inhibitory effect on the growth of L. monocytogenes, E. coli, and S. aureus during 12 h of incubation, and the 8 mM suspensions of ZnO were the most effective against all the strains. These data suggested that the antibacterial activity of ZnO was concentration dependent. Thus, 6 and 8 mM ZnO were selected for further studies in meat. ZnO nanoparticles reduced initial growth of all inoculated strains in meat. To our knowledge, this is the first report describing the antibacterial activity of ZnO nanoparticles in meat and indicates the potential of these nanoparticles as an antibacterial agent in the food industry.


2020 ◽  
Author(s):  
Giriraj Tailor ◽  
Jyoti Chaudhary ◽  
Ajit Joshi ◽  
Deepshikha Verma ◽  
Osahon Michael

Abstract The bioactive chromium nanoparticles were synthesized by calcination followed by thermal decomposition method. The antibacterial activity of chromium nanoparticles diffused in Dimethyl sulphoxide (DMSO). The antibacterial activity of chromium nanoparticles carried out against significant human pathogens (gram negative bacteria) viz, K. pneumonia, E. coli and P. typhus using agar diffusion cup plate method at 100 µg/ml concentration. The highest zone of inhibition was observed (12.0 mm) against K. pneumonia and lowest zone of inhibition (7.0 mm) E. coli. Thus, the outcomes of these studies suggest that synthesized chromium nanoparticles are of clinical importance.


2014 ◽  
Vol 1 (1) ◽  
pp. 5
Author(s):  
Umasankar Kulandaivelu ◽  
Bhawatha Chawada ◽  
Shireesha Boyapati ◽  
Alavala Rajasekhar Reddy

Arylalkylidene derivatives of aminotriazoles (3a-3j) were synthesized and tested for their antimicrobial and anticancer activity. Four non-pathogenic bacteria [E. coli (NCIM 2068), K. pneumoniae (NCIM 2957), S. aureus (NCIM 2079), B. subtilis (NCIM 2921)] two fungi [C. albicans, A. niger] and two cancer cell lines [HBL-100 and HT-29] were employed in the study. All the compounds were found to have better antibacterial activity against B. subtilis than Ciprofloxacin (standard) and compound 3i was equivalent to Ciprofloxacin in inhibiting S. aureas. Similarly all the compounds inhibited the growth of A. niger better than Fluconazole and compound 3c was equivalent to Fluconazole (standard) in inhibiting C. albicans. In case of anticancer activity none of the molecule exhibited activity better than the standard used (Methotrexate), though they have inhibitory concentration at submicromolar level.


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