The document summarizes a study that investigated the sensitivity of Pseudomonas species expressing Extended Spectrum Beta Lactamase (ESBL) to different solvent fractions of the plant Milletia aboensis.
Key findings:
- 10 isolates of Pseudomonas species expressing ESBL were obtained from animal samples.
- The ethanol, chloroform and methanol fractions of M. aboensis showed antibacterial activity against the ESBL-producing Pseudomonas isolates. The ethanol extract had the lowest minimum inhibitory concentration.
- The ethyl acetate fraction did not show significant antibacterial effects.
The results suggest M. aboensis contains anti-pseudomonal agents that could treat infections caused by
Invitro Evaluation of Antimicrobial Activity of the plant extracts of Elytrar...IJERA Editor
Antimicrobial activity of the aerial parts of the Elytraria acaulis a stem less perennial herb of Acantheceae family has been carried out in the present study. Extracts of the aerial parts of the plant (Stem & Leaves) were prepared in different organic solvents such as n-Hexane, Ethanol, Methanol and Ethyl acetate. All the extracts were analyzed for antimicrobial properties against various pathogenic bacterial infections (Escherichia coli, Klebsiella Species, and Salmonella Typhi). Amongst all the extracts, methanolic extract exhibited significant antimicrobial activity. The crude methanol extracts of leaves showed good inhibitory effects against pathogens. Hence further studies on this plant will enable elucidation of its therapeutic properties and medicinal applications.
Effect of Agrobacterium Induced Necrosis, Antibiotic Induced Phytotoxicity an...Sandip Magdum
Agrobacterium tumefaciens infection and antibiotic wash are the critical steps of Agrobacterium mediated plant transformation procedure, most time responsible for lower transformation efficiency due to necrosis and phytotoxicity caused by biotic stress of Agrobacterium and abiotic stress by antibiotics respectively. Ammi majus Egyptian origin medicinal plant and Pearl millet cereal grain crop were studied for their stress responses to Agrobacterium mediated transformation (AMT). Agrobacterium strains LBA4404 (O.D.=0.6-0.8) and EHA105 (O.D.=0.2-0.4) were used for transformation experiments to infect calli of Ammi majus and embryogenic calli of Pearl millet respectively. Incase of antibiotic wash, Cefotaxime 500 mg L-1 was used for LBA4404 infected Ammi majus calli and Timentin 300 mg L-1 was used for EHA105 infected embryogenic calli of Pearl millet.
Effects of Agrobacterium infection, antibiotic and NaOCl washes on Agrobacterium removal and both explants physiological changes during transformation experimental procedures were studied. At the end of the experiments explants survival efficiency of Ammi majus and pearl millet were 8% and 5% respectively. Biotic and abiotic stress factors responsible for lower efficiency were investigated with various other factors and strategies were discussed which are need to be considered for higher transformation events and target tissue survival.
Invitro Evaluation of Antimicrobial Activity of the plant extracts of Elytrar...IJERA Editor
Antimicrobial activity of the aerial parts of the Elytraria acaulis a stem less perennial herb of Acantheceae family has been carried out in the present study. Extracts of the aerial parts of the plant (Stem & Leaves) were prepared in different organic solvents such as n-Hexane, Ethanol, Methanol and Ethyl acetate. All the extracts were analyzed for antimicrobial properties against various pathogenic bacterial infections (Escherichia coli, Klebsiella Species, and Salmonella Typhi). Amongst all the extracts, methanolic extract exhibited significant antimicrobial activity. The crude methanol extracts of leaves showed good inhibitory effects against pathogens. Hence further studies on this plant will enable elucidation of its therapeutic properties and medicinal applications.
Effect of Agrobacterium Induced Necrosis, Antibiotic Induced Phytotoxicity an...Sandip Magdum
Agrobacterium tumefaciens infection and antibiotic wash are the critical steps of Agrobacterium mediated plant transformation procedure, most time responsible for lower transformation efficiency due to necrosis and phytotoxicity caused by biotic stress of Agrobacterium and abiotic stress by antibiotics respectively. Ammi majus Egyptian origin medicinal plant and Pearl millet cereal grain crop were studied for their stress responses to Agrobacterium mediated transformation (AMT). Agrobacterium strains LBA4404 (O.D.=0.6-0.8) and EHA105 (O.D.=0.2-0.4) were used for transformation experiments to infect calli of Ammi majus and embryogenic calli of Pearl millet respectively. Incase of antibiotic wash, Cefotaxime 500 mg L-1 was used for LBA4404 infected Ammi majus calli and Timentin 300 mg L-1 was used for EHA105 infected embryogenic calli of Pearl millet.
Effects of Agrobacterium infection, antibiotic and NaOCl washes on Agrobacterium removal and both explants physiological changes during transformation experimental procedures were studied. At the end of the experiments explants survival efficiency of Ammi majus and pearl millet were 8% and 5% respectively. Biotic and abiotic stress factors responsible for lower efficiency were investigated with various other factors and strategies were discussed which are need to be considered for higher transformation events and target tissue survival.
Biodegradation of Profenofos Pesticide by Efficient Bacillus Cereus and Klebs...ijsrd.com
The objective of this study to examine potential for the degradation Profenofos pesticide by the bacteria and finding the optimum conditions of bacteria. The growth of the pesticide degrading bacteria was assessed in Mineral salt broth containing 25mg of pesticide at different level temperature levels (25°C,30°C, 35°C & 40°C) and pH levels ( pH 5, pH 6, pH 7 & pH 8) .The maximum growth rate of bacteria was recorded at 35°C and pH 6. Among the tow bacteria the bacteria Bacillus cereus utilized the pesticides effectively and showed maximum growth. Profenofos pesticide was biological degradable.
Role of antimicrobial peptides in plant disease management N.H. Shankar Reddy
It is one of the advanced topics in plant disease management, detailed information about antimicrobial peptides and their role in plant disease management is furnished clearly.
Answering the Call to Arms: Tools for assessing the anti-infective potential ...Cassandra Quave
This is a presentation delivered at the 16th Annual Conference on the Science of Botanicals and 5th Annual Interim American Society of Pharmacognosy Meeting from April 11-14, 2016 in Oxford, MS, USA.
Abstract:
Answering the Call to Arms: Tools for Assessing the Anti-infective Potential of Natural Products in a Time of Rising Antibiotic Resistance
Quave CL1,2
1 Center for the Study of Human Health, Emory University, 550 Asbury Circle, Candler Library 107, Atlanta, GA 30322 USA. 2 Department of Dermatology, Emory University School of Medicine, 615 Michael Street, Whitehead 105L, Atlanta, GA 30322 USA.
As antibiotic resistance continues to rise, the pool of viable anti-infective therapeutic options is becoming rapidly exhausted. New therapies are in high demand and natural products are a likely source of novel bioactive compounds to meet this need. In particular, botanical secondary metabolites represent a rich pool for antibiotic discovery efforts. Plants are often the primary ingredients used in traditional anti-infective therapies, and yet their activity and mechanisms of action are often poorly understood. Much of the antibacterial research on botanical extracts and essential oils has focused on growth inhibitory studies using outdated methods limited in their ability to obtain an accurate assessment of bioactivity. The emergence of new molecular and bioanalytical tools for drug discovery provides a unique opportunity for application to natural products research.
Using Staphylococcus aureus as a model, tools for anti-infective testing of plant extracts will be reviewed, specifically focusing on the merits and limitations of each method. Examples include standardized methods for examining activity for the inhibition of growth (e.g., MIC, MBC), virulence (e.g., quorum sensing and toxin quantification) and pathogenesis (e.g., biofilms and antibiotic synergy). Data from our recent discoveries of novel biofilm [1] and quorum sensing [2,3] inhibitors isolated from medicinal plants (Rubus ulmifolius, Castanea sativa and Schinus terebinthifolius) will be presented in the review of these tools.
Acknowledgements: This work was supported by a grant from the National Institutes of Health, National Center for Complementary and Integrative Health (R01 AT007052). The content is solely the responsibility of the authors and does not necessarily reflect the official views of NCCIH or NIH.
References: [1] Quave CL, Estévez-Carmona M, et al. (2012) PLoS ONE, 7(1): e28737. [2] Quave CL, Lyles JT, et al. (2015) PLoS ONE, 10(8): e0136486. [3] Quave CL, Horswill AR (2014) Frontiers in Microbiology, 5: 706.
The present study aims to (I) evaluate the antiviral activity of eugenol oil nanoemulsion (EON) on eliminate Banana bunchy top virus (BBTV) from naturally infected banana plants and produce virus-free banana plants, (II) identify fungal contaminants of in vitro banana cultures and (III) evaluate the potential of EON on the suppression of the identified microbial contaminants and reduce of their occurrence frequency.
— The microbiological content of Lettuce (a vegetable), commonly vended in the Benin metropolis of Edo state were evaluated. Five vending locations were chosen for the study. Whole and soft rot samples were purchased and analysed for microbiological composition. Results showed high counts in soft rot samples in lettuce. Nutrient agar plated lettuce samples had bacterial counts in the range of 2.0x 103 to 4.7x10 7. Pseudomonas species was the dominant species found in lettuce samples. Bacillus species was isolated from one location in the lettuce samples. Mac Conkey agar plated lettuce plated had bacterial counts in the range of 2.3 x 10 3 to 5.7x 10 7. Enterobacter species, E. coli, and Klebsiella species were the dominant species isolated. Though, Proteus species was isolated from lettuce samples obtained from location five only. The study observes that consuming soft rot samples could pose a risk of introducing pathogens to the consumer due to their high microbial counts and could be detrimental to the health of the consumer.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
Biodegradation of Profenofos Pesticide by Efficient Bacillus Cereus and Klebs...ijsrd.com
The objective of this study to examine potential for the degradation Profenofos pesticide by the bacteria and finding the optimum conditions of bacteria. The growth of the pesticide degrading bacteria was assessed in Mineral salt broth containing 25mg of pesticide at different level temperature levels (25°C,30°C, 35°C & 40°C) and pH levels ( pH 5, pH 6, pH 7 & pH 8) .The maximum growth rate of bacteria was recorded at 35°C and pH 6. Among the tow bacteria the bacteria Bacillus cereus utilized the pesticides effectively and showed maximum growth. Profenofos pesticide was biological degradable.
Role of antimicrobial peptides in plant disease management N.H. Shankar Reddy
It is one of the advanced topics in plant disease management, detailed information about antimicrobial peptides and their role in plant disease management is furnished clearly.
Answering the Call to Arms: Tools for assessing the anti-infective potential ...Cassandra Quave
This is a presentation delivered at the 16th Annual Conference on the Science of Botanicals and 5th Annual Interim American Society of Pharmacognosy Meeting from April 11-14, 2016 in Oxford, MS, USA.
Abstract:
Answering the Call to Arms: Tools for Assessing the Anti-infective Potential of Natural Products in a Time of Rising Antibiotic Resistance
Quave CL1,2
1 Center for the Study of Human Health, Emory University, 550 Asbury Circle, Candler Library 107, Atlanta, GA 30322 USA. 2 Department of Dermatology, Emory University School of Medicine, 615 Michael Street, Whitehead 105L, Atlanta, GA 30322 USA.
As antibiotic resistance continues to rise, the pool of viable anti-infective therapeutic options is becoming rapidly exhausted. New therapies are in high demand and natural products are a likely source of novel bioactive compounds to meet this need. In particular, botanical secondary metabolites represent a rich pool for antibiotic discovery efforts. Plants are often the primary ingredients used in traditional anti-infective therapies, and yet their activity and mechanisms of action are often poorly understood. Much of the antibacterial research on botanical extracts and essential oils has focused on growth inhibitory studies using outdated methods limited in their ability to obtain an accurate assessment of bioactivity. The emergence of new molecular and bioanalytical tools for drug discovery provides a unique opportunity for application to natural products research.
Using Staphylococcus aureus as a model, tools for anti-infective testing of plant extracts will be reviewed, specifically focusing on the merits and limitations of each method. Examples include standardized methods for examining activity for the inhibition of growth (e.g., MIC, MBC), virulence (e.g., quorum sensing and toxin quantification) and pathogenesis (e.g., biofilms and antibiotic synergy). Data from our recent discoveries of novel biofilm [1] and quorum sensing [2,3] inhibitors isolated from medicinal plants (Rubus ulmifolius, Castanea sativa and Schinus terebinthifolius) will be presented in the review of these tools.
Acknowledgements: This work was supported by a grant from the National Institutes of Health, National Center for Complementary and Integrative Health (R01 AT007052). The content is solely the responsibility of the authors and does not necessarily reflect the official views of NCCIH or NIH.
References: [1] Quave CL, Estévez-Carmona M, et al. (2012) PLoS ONE, 7(1): e28737. [2] Quave CL, Lyles JT, et al. (2015) PLoS ONE, 10(8): e0136486. [3] Quave CL, Horswill AR (2014) Frontiers in Microbiology, 5: 706.
The present study aims to (I) evaluate the antiviral activity of eugenol oil nanoemulsion (EON) on eliminate Banana bunchy top virus (BBTV) from naturally infected banana plants and produce virus-free banana plants, (II) identify fungal contaminants of in vitro banana cultures and (III) evaluate the potential of EON on the suppression of the identified microbial contaminants and reduce of their occurrence frequency.
— The microbiological content of Lettuce (a vegetable), commonly vended in the Benin metropolis of Edo state were evaluated. Five vending locations were chosen for the study. Whole and soft rot samples were purchased and analysed for microbiological composition. Results showed high counts in soft rot samples in lettuce. Nutrient agar plated lettuce samples had bacterial counts in the range of 2.0x 103 to 4.7x10 7. Pseudomonas species was the dominant species found in lettuce samples. Bacillus species was isolated from one location in the lettuce samples. Mac Conkey agar plated lettuce plated had bacterial counts in the range of 2.3 x 10 3 to 5.7x 10 7. Enterobacter species, E. coli, and Klebsiella species were the dominant species isolated. Though, Proteus species was isolated from lettuce samples obtained from location five only. The study observes that consuming soft rot samples could pose a risk of introducing pathogens to the consumer due to their high microbial counts and could be detrimental to the health of the consumer.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
In vitro controlling of selected human diarrhea causing bacteria by clove ext...Open Access Research Paper
Antibacterial activity of clove extracts (Syzygium aromaticum L.) was proven against five diarrhea causing bacteria. This was further confirmed when compared with commonly used three commercial antibiotics (ciprofloxacin, tetracycline and erythromycin) as a positive control. Significant differences (P<0.0001) were observed in the effect of the antimicrobial agents (clove extracts and antibiotics), and in the sensitivities of the bacterial species (P<0.0001) to the antimicrobial agents. Clove extracts had significant (P<0.001) activity with the acetone extract demonstrating highest activity followed by antibiotics and other extracts against tested bacteria. The zone of inhibition of clove extracts was ranged from 7.33 to 12.00 mm whereas in antibiotics, it was 0.00 to 11.67 mm. Of all the bacteria, Salmonella typhimurium was the most susceptible against all of the extracts as well as concentrations of clove, while low MIC (180 mgml-1) and MBC (680 mgml-1) of the extracts were observed against Shigella dysenteriae. Consequently, clove has a significant antidiarrheal activity and it could be used as an effective antibacterial agent, alternative to the use of antibiotics.
Antibiotic Susceptibility Pattern of Pyogenic Bacterial Isolates in Sputum.IOSR Journals
Drugs Have Been Used For The Treatment Of Infectious Diseases Since 17th Century , However
Chemotherapy As A Science Has Began With Paul Ehrlich In The First Decade Of 20th Century . Paul Ehrlich
(1854-1915) Was One Of The Earliest Pioneers In The Field Of Antimicrobial Chemotherapy .1Ehrlich
Formulated The Principles Of “Selective Toxicity” ,I.E; Selective Inhibition Of The Growth Of Microorganisms
Without Damage To The Host.2 Resistance Has Been Documented Not Only Against Antibiotics Of
Natural And Semi- Synthetic Origin , But Also Against Purely Synthetic Compounds (Flouroquinolone) Or
Those Which Do Not Even Enter The Cells (Vancomycin) .3 However , The Euphoria Over The Potential
Conquest Of Infectious Diseases Was Short-Lived .Almost As Soon As Antibacterial Drugs Were Deployed ,
Bacteria Responded By Manifesting Various Forms Of Resistance.4 Considered As “Wonder Drugs”
Antibiotics Are Often Prescribed Inappropriately And Inadequately And Have Thus Became One Of The
Highly Abused Agents.5
“Isolation and Biochemical Characterization of Antibiotic Producing Microorga...IOSR Journals
The search for new antibiotics continues in a rather overlooked hunting ground. In the course of screening for new antibiotic-producing microorganisms, isolates showing antimicrobial activity were isolated from waste soil samples from various habitats in the Industrial Areas in Dheradun, Uttarakhand, India. Existing methods of screening for antibiotic producers together with some novel procedures were reviewed. Both modified agar-streak and agar-plug methods were used in the primary screens. The use of selective isolation media, with or without antibiotic incorporation and/or heat pretreatment, enhanced the development of certain actinomycete colonies on the isolation plates. Antibiotics have long been considered the “magic bullet” that would end infectious disease. Although they have improved the health of countless numbers of humans and animals, many antibiotics have also been losing their effectiveness since the beginning of the antibiotic era. Bacteria have adapted defenses against these antibiotics and continue to develop new resistances, even as we develop new antibiotics. In recent years, much attention has been given to the increase in antibiotic resistance. As more microbial species and strains become resistant, many diseases have become difficult to treat, a phenomenon frequently ascribed to both indiscriminate and inappropriate use of antibiotics in human medicine. However, the use of antibiotics and antimicrobials in raising food animals has also contributed significantly to the pool of antibiotic resistant organisms globally and antibiotic resistant bacteria are now found in large numbers in virtually every ecosystem on earth. Dual culture bioassays were used to screen seven selected Bacillus isolates for activity against four plant pathogenic fungi in vitro. All isolates were able to inhibit the pathogens to varying degrees. Two isolates, R29 and B81, were selected for further testing and characterization. Further bioassays were performed on five complex nutrient media which were adjusted to pH S.S and 7, and both incubated at 2SoC and 30°C" respectively. It was found that pH and media composition showed significant influences on the antifungal activities of the isolates tested, but that a SoC temperature difference in incubation temperature did not. Tryptone soy agar was found to give rise to the largest inhibition zones. Both isolates were tentatively identified using standard biochemical and morphological tests. Based on its phenotypic characteristics, R29 was identified as a strain of B. subtilis. B81 proved to be more difficult to assign to a specific group or species of Bacillus, though B. subtilis and B. licheniformis were considered to be the nearest candidates. Genomic DNA was extracted from both isolates and a portion of each of their 16s rDNA genes were amplified and sequenced for homology testing against the GeneBank database. Homology testing confirmed that both isolates were members of the genus Bacillus and most
In vitro Antimicrobial Activity Screening of Rheum rhabarbarum Rootsinventionjournals
Rheum rhabarbarum, which is commonly known as rhubarb, has been used as a medicinal herb in different countries. Especially its roots are known to be a traditional medicine in different cultures. Mesir paste was prepared about 500 years ago during Ottoman period as a medicinal paste and R. rhabarbarum was one of its ingredients. In this study the in vitro antimicrobial activity of ethanol extract of R. rhabarbarum roots was investigated against 17 bacterial and 1 fungal strain, namely, Bacillus subtilis DSMZ 1971, Candida albicans DSMZ 1386, Enterobacter aerogenes ATCC 13048, Enterococcus durans, Enterococcus faecalis ATCC 29212, Enterococcus faecium, Escherichia coli ATCC 25922, Klebsiella pneumoniae, Listeria innocula, Listeria monocytogenes ATCC 7644, Pseudomonas aeruginosa DSMZ 50071, Pseudomonas fluorescence P1, Salmonella enteritidis ATCC 13075, Salmonella infantis, Salmonella kentucky, Salmonella typhimurium SL 1344, Staphylococcus aureus ATCC 25923 and Staphylococcus epidermidis DSMZ 20044 by using the disk diffusion method. It is observed that ethanol extracts of R. rhabarbarum root extracts has antimicrobial activity against all microorganims tested.
In vitro Antimicrobial Activity Screening of Rheum rhabarbarum Rootsinventionjournals
Rheum rhabarbarum, which is commonly known as rhubarb, has been used as a medicinal herb in different countries. Especially its roots are known to be a traditional medicine in different cultures. Mesir paste was prepared about 500 years ago during Ottoman period as a medicinal paste and R. rhabarbarum was one of its ingredients. In this study the in vitro antimicrobial activity of ethanol extract of R. rhabarbarum roots was investigated against 17 bacterial and 1 fungal strain, namely, Bacillus subtilis DSMZ 1971, Candida albicans DSMZ 1386, Enterobacter aerogenes ATCC 13048, Enterococcus durans, Enterococcus faecalis ATCC 29212, Enterococcus faecium, Escherichia coli ATCC 25922, Klebsiella pneumoniae, Listeria innocula, Listeria monocytogenes ATCC 7644, Pseudomonas aeruginosa DSMZ 50071, Pseudomonas fluorescence P1, Salmonella enteritidis ATCC 13075, Salmonella infantis, Salmonella kentucky, Salmonella typhimurium SL 1344, Staphylococcus aureus ATCC 25923 and Staphylococcus epidermidis DSMZ 20044 by using the disk diffusion method. It is observed that ethanol extracts of R. rhabarbarum root extracts has antimicrobial activity against all microorganims tested
Antibacterial Activity of Leaf Methanolic Extract of S. Caryophyllatum (L.) A...iosrjce
S.caryophyllatum (L.) Alston belongs to the family Myrtaceae is an endangered species. It possesses
traditional as well as pharmacological properties. The objective of the present investigation was to find out the
antibacterial activity of S. caryophyllatum leaf methanolic extract against some human pathogenic bacteria. It
was followed by Disc Diffusion method using gram positive and gram negative bacterial strains such as
Staphylococcus aureus, Bacillus cereus, Bacillus subtilis, Bacillus megaterium, Sarcina lutea, Esherichia coli,
Pseudomonas aeruginosa, Klebsiella Spp., Salmonella typhi and Proteus mirabilis. The result showed that the
inhibitory effect on Bacillus subtilis (24mm) was high when compared to E. coli (21mm) and Bacillus cereus
(20mm).This effect on the bacterial strains may be due to the presence of secondary metabolites present in the
leaf methanolic extract of Syzygium caryophyllatum.
Synergistic antibacterial effects of three edible plants extract against anti...Open Access Research Paper
In vitro synergistic antibacterial effects among Alocasia macrorrhizos rhizome, Amorphophallus paeoniifolius corm and Colocasia esculenta corm extracts were tested against six resistant bacteria viz., Yersinia enterocolitica, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Clostridium difficile and Staphylococcus aureus. The inhibition zone was compared with the commercially available antibiotic (tetracycline). High inhibitory activity was observed against E. coli (12.67±0.33 mm) and S. aureus (12.50±0.29 mm) for methanol extract at 800 mgml-1 of concentration. MIC and MBC of the extracts ranged from 200-580 mgml-1 and 250-650 mgml-1 respectively. The lowest MIC and MBC of the extracts were measured against E. coli.
Antibiogram of Staphylococcus Aureus and its Sensitivity to Ocimum Gratissimu...ijtsrd
In this study, clinical isolate of Staphylococcus aureus was subjected to susceptibility tests against commercially available antibiotics and Ocimum gratissimum scent leaf leaf extracts. disc diffusion nmethod was adopted for the antibiotics test while agar well diffusion technique was employed for the antimicrobial screening of the leaf extracts. The result shows that the organism was sensitive to Gentamycin, Zinnacef, Ciprofloxacin and Streptomycin representing 40 of the antibiotics while showing outright resistance to Ampiclox, Amoxicillin and Erythromycin 30 and intermediate results for the rest. Also, the cold water extract of O. gratissimum did not show any activity against the bacterium. However, both hot water extract and ethanol extract of the plant had varying degree of activity against the organism with ethanol extract recording a zone of inhibition of 25.33 mm compared with 32.00 mm recorded for Gentamycin. Furthermore, the minimum inhibitory concentration recorded ofr the extract against the organism were 10 mg ml and 2.5 mg ml respectively for hot water and ethanol extracts. These results indicate that ethanol extract of O. gratissimum leaf is very effective against Staphylococcus aureus and could be used as potential source of natural product for the treatment of infections caused by the organism especially the antibiotic resistant strains. Komolafe T. O. | Ogunyankin O. G "Antibiogram of Staphylococcus Aureus and its Sensitivity to Ocimum Gratissimum Extract" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-6 , October 2021, URL: https://www.ijtsrd.com/papers/ijtsrd46333.pdf Paper URL : https://www.ijtsrd.com/other-scientific-research-area/other/46333/antibiogram-of-staphylococcus-aureus-and-its-sensitivity-to-ocimum-gratissimum-extract/komolafe-t-o
Invitro Study of Antibacterial Activity of Leaf and Root Extract of Rauvolfia...paperpublications3
Abstract: In this study Methanolic and chloroform leaf and root extract of Rauvolfia serpentina was studied for its antibacterial activity. Antibacterial activity of leaf and root extracts was assessed against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Klebsiella pneumonia by disc diffusion method. Methanolic extract of root was showed the maximum zone of inhibition for all test organisms than the leaf extract. According to observations of root extract of 50µl/ml concentration 15.4mm, 16.2mm, 12.3mm,10.1mm and 15.0mm zones of inhibition and for concentration of 100µl/ml 22.5mm, 23.1mm, 15.1mm, 18.0mm, 22.0mm zones of inhibition were formed against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Klebsiella pneumonia respectively. 50µl/ml concentration of leaf and root chloroform extracts showed no zone of inhibition against Staphylococcus aureus and Bacillus subtilis, maximum zone of inhibition was observed 15.0mm and 15.5mm against E. coli for leaf and root chloroform extract respectively. 100µl/ml concentration showed maximum zone of inhibition against all test organisms for both leaf and root extracts. All the bacteria were more susceptible to methanolic extracts than the chloroform extracts.
The International Journal of Engineering and Science (The IJES)theijes
The International Journal of Engineering & Science is aimed at providing a platform for researchers, engineers, scientists, or educators to publish their original research results, to exchange new ideas, to disseminate information in innovative designs, engineering experiences and technological skills. It is also the Journal's objective to promote engineering and technology education. All papers submitted to the Journal will be blind peer-reviewed. Only original articles will be published.
The papers for publication in The International Journal of Engineering& Science are selected through rigorous peer reviews to ensure originality, timeliness, relevance, and readability
ANTIOXIDANT AND ANTIMICROBIAL ACTIVITIES OF ALGERIAN POPULUS NIGRA L. BUDS EX...bioejjournal
This study is part of a goal to investigate chemical composition, antibacterial, antifungal and antioxidant
activities of the flower buds extracts from the Algerian Polulus nigra L., which were collected from Djarifet
- mansourah at Tlemcen city in the West Northern of Algeria.
In organic extracts, tanins, flavonoïds, coumarins, alkaloids and terpenoïds were the principals secondary
metabolites identified from the flower buds of black poplar. Antibacterial and antifungal activities of
extracts were tested using agar-well diffusion method and micro-well determination of MIC assay against
eleven bacteria and two Candida species. It was found that extracts of black poplar buds exhibit
antibacterial and anticandidal activities with agar disk diffusion (7 to 43mm) and MIC methods (MIC=
90.33 μg/ml against several strains of bacteria and MIC=45.16 μg/ml against Candida albicans). The
antioxidant effect of hydroalcoholic extract was evaluated using DPPH and FRAP assays. It was showed
good and similar activity than ascorbic acid and BHA by DPPH method: IC50= 220μg/mL for
hydroethanol extract.
Antioxidant and Antimicrobial Activities Of Algerian Populus Nigra L. Buds Ex...bioejjournal
This study is part of a goal to investigate chemical composition, antibacterial, antifungal and antioxidant activities of the flower buds extracts from the Algerian Polulus nigra L., which were collected from Djarifet - mansourah at Tlemcen city in the West Northern of Algeria. In organic extracts, tanins, flavonoïds, coumarins, alkaloids and terpenoïds were the principals secondary metabolites identified from the flower buds of black poplar. Antibacterial and antifungal activities of
extracts were tested using agar-well diffusion method and micro-well determination of MIC assay against
eleven bacteria and two Candida species. It was found that extracts of black poplar buds exhibit
antibacterial and anticandidal activities with agar disk diffusion (7 to 43mm) and MIC methods (MIC=
90.33 µg/ml against several strains of bacteria and MIC=45.16 µg/ml against Candida albicans). The
antioxidant effect of hydroalcoholic extract was evaluated using DPPH and FRAP assays. It was showed good and similar activity than ascorbic acid and BHA by DPPH method: IC50= 220µg/mL for hydroethanol extract.
ANTIOXIDANT AND ANTIMICROBIAL ACTIVITIES OF ALGERIAN POPULUS NIGRA L. BUDS EX...bioejjournal
his study is part of a goal to investigate chemical composition, antibacterial, antifungal and antioxidant activities of the flower buds extracts from the Algerian Polulus nigra L., which were collected from Djarifet - mansourah at Tlemcen city in the West Northern of Algeria. In organic extracts, tanins, flavonoïds, coumarins, alkaloids and terpenoïds were the principals secondary metabolites identified from the flower buds of black poplar. Antibacterial and antifungal activities of extracts were tested using agar-well diffusion method and micro-well determination of MIC assay against eleven bacteria and two Candida species. It was found that extracts of black poplar buds exhibit antibacterial and anticandidal activities with agar disk diffusion (7 to 43mm) and MIC methods (MIC= 90.33 µg/ml against several strains of bacteria and MIC=45.16 µg/ml against Candida albicans). The antioxidant effect of hydroalcoholic extract was evaluated using DPPH and FRAP assays. It was showed good and similar activity than ascorbic acid and BHA by DPPH method: IC50= 220µg/mL for hydroethanol extract.
Bioactivity screening of Soil bacteria against human pathogenspharmaindexing
Microorganisms have a profound effect on medical science as they not only infect & cause disease but also produce metabolic products that can cure infections. Soil happens to be a source for a variety of microorganisms. Most of the bacteria, particularly actinomycetes produce biologically active secondary metabolites. Though there are a number of antibiotics available, there is a pressing need for the discovery of new source for antimicrobials against the pathogens due to the development of drug resistance of the pathogenic microorganisms. In addition to, new pathogenic strains are also developing and causing infection to human beings. Bioactive compounds are compounds that are produced by any living organism and are known to exhibit various biological activities both in-vitro & in-vivo. Bioactivity may be antimicrobial, antineoplastic, anticancerous, immunomodulation, antifertility & others. Soil bacteria were isolated by standard technique and by making use of selective media. The isolates were identified and subjected for preliminary screening to look for their ability to produce bioactive materials. A total of 96 strains were isolated from three different soil samples. 14 of them were found to have antibacterial activity against the human pathogens like Staphylococcus aureus, Streptococcus faecalis, E.coli, Klebsiella aerogenes, Proteus vulgaris, Pseudomonas aureginosa and Salmonella typhi by preliminary screening. Further the selected (3) bacteria were grown in the suitable culture media for the production of bioactive metabolites by using rotary shake flask. The active metabolites was isolated by solvent extraction and concentrated by evaporation under reduced pressure. The antimicrobial screening of the active metabolites showed prominent effect against the clinical pathogens under the study.
Sensitivity of pseudomonas species expressing extended spectrum
1. Journal of Natural Sciences Research www.iiste.org
ISSN 2224-3186 (Paper) ISSN 2225-0921 (Online)
Vol.2, No.9, 2012
Sensitivity of Pseudomonas Species Expressing Extended Spectrum
Beta Lactamase to Different Solvent Fractions of Milletia Aboensis
Moses N Ikegbunam1, Calistus D Nwakile2*, Malachy C Ugwu1, Daniel L Ajaghaku3 and Charles O Esimone1
1. Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences,
Nnamdi Azikiwe University Awka Anambra State Nigeria
2. Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Nnamdi
Azikiwe University Awka Anambra State Nigeria
3. Department of Pharmacology and Toxicology, Faculty of Pharmaceutical Sciences, Nnamdi Azikiwe
University Awka Anambra State Nigeria
* Email of the corresponding author:cdnwakile@gmail.com
Abstract
The presence of Extended Spectrum Beta Lactamase (ESBL) producing organisms in abattoirs, a non-hospital
community was investigated. A total of ten (10) isolates of Pseudomonas species out of twenty-six bacteria isolates
expressing ESBL was obtained. The anti-pseudomonal activities of various solvent fractions of Milletia aboensis
against the ESBL positive isolates of Pseudomonas species showed varying sensitivity. These results have suggested
that Milletia aboensis possess potent anti-pseudomonal agents that could be used to treat infections due to
Pseudomonas species expressing ESBL. These anti-pseudomonal metabolites are located in the ethanol, chloroform
and methanol fractions but are absent in the ethyl acetate fractions.
Keywords: Extended Spectrum Beta Lactamase, Milletia aboensis, Pseudomonas species
1.0 Introduction
The emergence of antibiotic resistance among prevalent pathogens is a serious threat to the management of
infectious disease. Beta-lactam (β-lactam) antibiotics are commonly used for the treatment of bacterial infections [1].
However the development of resistance to this class of antibiotics by bacteria through production of β-lactamases is
the main mechanism of bacterial resistance to these antibiotics [2]. The extended-spectrum β-lactams became widely
used in the treatment of serious infection due to Gram-negative bacteria [3] after failure of the Beta-lactams. However,
resistance to these extended spectrum β-lactams due to β-lactamases emerged quickly [3, 4]. The first report of
plasmid-encoded β-lactamases capable of hydrolyzing the extended-Spectrum cephalosporins was published in
1983[5]. Many Gram-negative bacteria possess naturally occurring, chromosomally mediated-lactamases, (e.g. AmpC
cephalosporinases of Enterobacteriaceae) as well as plasmid-mediated β-lactamase e.g. TEM-1 and SHV-1[2] that
inactivate these antibiotics. Extended Spectrum Beta Lactamases (ESBLs) are so called because they are able to
hydrolyze extended spectrum β-lactam antibiotics. They have ability to inactivate β-lactam antibiotics containing an
oxyimino-group such as oxyimino-cephalosporins (e.g., ceftazidime, ceftriaxone, cefotaxime) as well as oxyimino-
monobactam (e.g. aztreonam) [3, 6]. The enzymes are not active against cephamycins and carbapenems. Generally,
they are inhibited by β-lactamase-inhibitors such as clavulanate and tazobactam. ESBLs have been isolated in food
producing animals, Community and Hospital settings and among paediatrics [7-9].
Milletia aboensis is widely distributed in tropical Africa and found abundantly in South East of Nigeria, Cameroon
and Equatorial Guinea. The leaves are used for general healing and as laxatives while the root is used for treatment
of venereal diseases [7]. Traditionally, the leaves decoction has been widely used in the treatment of pains and
inflammatory disease conditions.
In this research therefore, the crude extracts and some solvent fractions of the crude extracts are evaluated against
ESBL producing Pseudomonas species. The findings will enhance the search for new active ingredient(s) for the
control of the ESBL producing pseudomonas species in particular and bacteria in general.
1.1 Materials and Methods
1.1.1 Isolation of ESBL producing Pseudomonas species using Double Disc Synergy Test (DDST)
A total of twenty-nine bacteria showing ESBL positivity isolated from One hundred and forty nine (149) samples
collected from different Cow abattoirs at Awka metropolis Anambra State Nigeria were used for the study.
A 20 ml volume of Mueller Hinton agar was prepared and dispensed aseptically into each of the Petri dishes. A 0.1
ml suspension of each of the isolates equivalent to 0.5 ml MacFarland standard was aseptically seeded into the Petri
dishes together with Mueller Hinton agar. This was allowed to stand for 1 hour to solidify. A combination disc
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ISSN 2224-3186 (Paper) ISSN 2225-0921 (Online)
Vol.2, No.9, 2012
(Amoxicillin 20 µg and Clavulanic acid 10 µg) was placed at the centre of the Petri dish and antibiotics (Ceftazidime
30 µg and Cefuroxime 30 µg) were placed 15 mm apart centre to centre on both sides of the plates.
The set up was done in triplicate and it was left for 30 minutes for diffusion. It was incubated at 37 oC for 24 hours
after which the various inhibition zone diameters were measured.
1.1.2 Extraction and fractionation of the plant materials
The fresh plant leaves were washed with clean water, then sliced into small fragments and sun-dried. The dried
material was reduced to coarse powder with mortar and pestle and then reduced to a fine powder with a mechanical
grinder.
About 500 g of the pulverized leaves were cold-macerated in aqueous ethanol (70%) for forty-eight hours. The
ethanol extract was concentrated in vacuum. The extract was adsorbed on silica gel and eluted in succession with
Chloroform, Ethyl acetate and Methanol. All the extracts and fractions were stored in refrigerator between 0-4.oC
until used.
1.1.3 Sensitivity of test microorganisms to plant extracts
This was determined by the agar well diffusion method. A 20 ml quantity of sterile molten Mueller- Hinton nutrient
agar was poured aseptically into Petri dishes containing 0.1 ml of the McFarland’s standard of the test micro-
organisms. The plates were gently rotated to ensure even distribution of the microorganisms and finally allowed to
set. Using sterile cork borer, wells of 6 mm diameter were bored on the solidified seeded Mueller-Hinton nutrient
agar plates. About 40 µl of the different concentrations of herbal extract and standard antibiotics were placed in the
wells and left for one hour at room temperature after which the plates were incubated at 37 oC for 24 hours. The test
was carried out in triplicate for each of the ten preparations. After the incubation period, the plates were observed for
inhibition zone diameter (IZD).
1.1.4 Evaluation of the Minimum Inhibitory Concentration (MIC) of the plant extract
The minimum inhibitory concentration of the extracts and standard antibiotics against Pseudomonas species
expressing ESBL were performed using the agar dilution method. A 19 ml volume of sterilized Mueller-Hinton
nutrient agar was aseptically poured into a sterile Petri dish containing 1 ml of the graded concentration of the eight
different preparations of the extracts. The plates were rotated to ensure an even distribution of the microorganisms
and allowed to set. The plates were inoculated with a loopful of 0.5 McFarland standards of the test microorganisms.
This was repeated for all the test microorganisms and each done in duplicates. The plates were incubated at 37 oC for
24 hours for all species of the ESBL producing Pseudomonas. Presence of growth was observed after incubation and
the MIC recorded as the minimum concentration where no visible growth was observed on the plate.
1.2 Results and Discussions
The ethanol extract of Milletia aboensis has a yield of 86.74 g. The chloroform fraction had a yield of 8.46 g, the
ethyl acetate fraction gave a yield of 5.48 g and that of methanol fraction was 35.46 g.
Phenotypic methods of detection of ESBL activity is based upon the resistance that ESBLs confer to oxyimino- β-
lactams (e.g ceftriaxone, cefotaxime, ceftazidime and aztreonam) and the ability of a β-lactamase inhibitor, usually
clavulanate, to block this resistance. A clear extension of the edge of the antibiotic’s inhibition zone toward the disk
containing clavulanate is interpreted as synergy, indicating the presence of an ESBL.
Table 1 thus shows high ESBL production with the organisms depicting the synergistic symbol that is characteristic
of ESBL production appearing to be Pseudomonas species. ESBLs have been found in a wide range of gram-
negative rods. However, the vast majority of strains expressing these enzymes belong to the family
Enterobacteriaceae [3]. Klebsiella pneumonia seems to remain the major ESBL producers. Another very important
organism is Escherichia coli. It is important to note the growing incidence of ESBLs in Salmonella sp [8]. ESBLs
have become more prevalent among species with inducible AmpC β- lactamases [9].
Non Enterobacteriaceae ESBL producers are relatively rare with Pseudomonas aeruginosa being the most
important organism [10]. ESBL has also been reported in Acinetobacter spp, Burkholderia cepacia and Alcaligenes
fecalis [3].
Some solvent fraction of Milletia aboensis showed an appreciable level of antibacterial activity on the Pseudomonas
spp producing ESBL (Table 2). Both the crude ethanol extract and methanol fraction showed appreciable activity
against ESBL Pseudomonas isolates at the concentration of 25mg/ml and 12.5mg/ml respectively (Table 2). It was
also observed that the activity increases as the concentration decreases which could be attributed to diffusion
problems. Also, table 2 revealed that the crude extract of Milletia aboensis had no effect against two species of
Pseudomonas (putida and fulva) while there are observable zones of inhibition when evaluated against P.monteilli,
P.beteli and P.mendocina. There are zones of inhibition across the entire organisms challenged with the methanol
extract of Milletia aboensis at the concentration ranges of 50mg/ml-12.5mg/ml.
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Chloroform fraction exhibited antibacterial activity at the highest concentration (Table 2) while ethyl acetate showed
no significant effect against the test ESBL isolates.
From table 3, the MIC of the Crude (ethanol) leaf extracts as well as the chloroform and methanol fractions of the
extracts of Milletia aboensis against the positive test ESBL isolates of Pseudomonas species were shown. The
ethanol extract had the lowest MIC value of 10 mg/ml. This could imply that the metabolites with highest activity
may lie within the ethanol extract or that group of metabolites act synergistically to give a better performance than
either the chloroform or methanol fractions.
Conclusion
The ESBL positive Pseudomonas species isolated are sensitive to metabolites found in the ethanol, chloroform and
methanol fractions of Milletia aboensis.
REFERENCES
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2. Livermore, D.M. (1995). β- lactamases in laboratory and clinical resistance. Clinical Microbiology Review,
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3. Bradford, P. A. (2001). Extended spectrum β-lactamases in the 21st century: characterization, epidemiology
and the detection of this important resistance threat, Clinical Microbiology Review, 14,933-951.
4. Samaha-Kfoury, J.N. and Araji, G. F. (2003). Recent developments in β- lactamases and extended spectrum
β- lactamases. Br Med Journal, 327, 1209-1213.
5. Knothe, H. et al (1983). Transferable resistance to cefotaxime, cefoxitin, cefamandole and Cefuroxime in
clinical isolates of Klebsiella pneumoniae and Serratia marcescens. Infection.11, 315 – 317.
6. Paterson, D.L and Bonomo, R.A. (2005). Extended spectrum Beta- lactamases: a clinical update. Clinical
Microbiology Review. 18, 657 -686.
7. Nadine Geser, Roger Stephan and Herbert Hächler (2012). Occurrence and characteristics of extended-
spectrum β-lactamase (ESBL) producing Enterobacteriaceae in food producing animals, minced meat and raw milk.
BMC Veterinary Research, 8:21 doi:10.1186/1746-6148-8-21.
8. Sun H Park, Ji-Hyun Byun, Su-Mi Choi, Dong-Gun Lee, Si-Hyun Kim, Jae-Cheol Kwon, Chulmin Park,
Jung-Hyun Choi and Jin-Hong Yoo (2012). Molecular epidemiology of extended-spectrum β-lactamase-producing
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Infectious Diseases, 12:149 doi: 10.1186/1471-2334-12-149
9. Yu F, Chen Q, Yu X, Li Q, Ding B, et al. (2011) High Prevalence of Extended-Spectrum Beta Lactamases
among Salmonella enterica Typhimurium Isolates from Pediatric Patients with Diarrhea in China. PLoS ONE 6(3):
e16801. doi:10.1371/journal.pone.0016801
10. Burkill, H.M. (1985). The Useful plants of West Tropical Africa. Vol 3 Jstor plant Science.
11. Windokur, P.L. et al (2000). Animal and human multidrug - resistant, cephalosporin – resistant Salmonella
isolates expressing a plasmid –mediated CMY -2 AmpC β-lactamase. Antimicrobial Agents Chemotherapy, 44,
2777-2783.
12. Levison, M.E. (2002). Plasmid-mediated extended –spectrum β-lactamases in organisms other than Klebsiella
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Table 1: Phenotypic confirmation of ESBL producing Pseudomonas spp
CAZ CAZ+AMC CTX CTX+AMC
P. monteilli 0 29 0 30
P. putida 21 28 0 22
P. beteli 17 28 0 28
P. mendocina 11 20 13 27
P. putida 15 28 0 21
P. monteilli 0 31 0 23
P. monteilli 15 33 0 22
P. putida 19 29 11 25
P. monteilli 0 30 13 29
P. fulva 21 34 0 24
P. putida 24 28 11 30
P. monteilli 0 31 0 24
P. fulva 21 29 14 29
P. mendocina 0 22 0 22
CAZ = Ceftazidime; CTX = Cefotaxime; AMC = Amoxicillin+ Clavulanic acid
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Table 3: Minimum inhibitory concentrations of solvent fractions of Milletia aboensis against
Pseudomonas species expressing extended spectrum beta lactamase
Isolates Minimum Inhibitory Concentration (mg/ml)
CEMA CFMA MFMA GEN (µg/ml)
B1 10 50 50 0.15
B2 10 50 50 0.15
B3 10 50 50 0.15
B4 10 50 50 0.15
B5 10 50 50 0.15
B6 10 50 50 0.15
B7 10 50 50 0.15
B8 10 50 50 0.15
B9 10 50 50 0.15
B10 10 50 50 0.15
*Each value represents the mean value of two determinants. B1= Pseudomonas monteilli, B2=P.
putida, B3=P.beteli, B4= P. mendocina, B5= P. putida, B6= P. monteilli, B7= P. monteilli, B8= P.
Putida, B9= P. fulva, B10= P. mendocina. CEMA= Crude Extract of Milletia aboensis, CFMA=
Chloroform Fraction of Milletia aboensis, MFMA= Methanolic Fraction of Milletia aboensis,
GEN = Gentamycin
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