1) The document reports on research into the endophytic fungus Papulaspora pallidula's ability to biosynthesize silver nanoparticles (AgNPs) and their potential antitumor and antibacterial efficacy.
2) The biosynthesized AgNPs exhibited 52.7% growth inhibition of human larynx carcinoma cells and varying levels of inhibition against 5 pathogenic bacterial strains. Combining the AgNPs with Gentamycin significantly increased antibacterial activity.
3) Characterization of the AgNPs showed they were spherical and ranged from 8-90nm in size. UV-Vis and FTIR analysis confirmed AgNP formation and protein capping, while SEM images visualized particle shape and dispersion.
The current research aimed at fabricating plant extract mediated biosynthesized silver nanoparticles (AgNPs) utilizing thorn extract of Bombax ceiba (TEBC). The synthesized AgNPs was characterized by UV spectroscopy where the surface plasmonic resonance peak (SPR) was located at 222 nm. The scanning electron microscopy (SEM) studies demonstrated that the morphology of fabricated nanomaterials was primarily cylindrical of average size of 20-30 nm with some spindles of size >50 nm. The anti-microbial evaluation against Staphylococcus aureus revealed that AgNPs exhibited notable activity with ZOI of 27.2 mm at MIC of 25 μg/mL. The outcome of this research evidently signified that the biofabricated AgNPs using TEBC may be a new greener approach or technology to formulate anti-bacterial nanodrugs in future.
Comparative study on screening methods of polyhydroxybutyrate (PHB) producing...inventionjournals
International Journal of Engineering and Science Invention (IJESI) is an international journal intended for professionals and researchers in all fields of computer science and electronics. IJESI publishes research articles and reviews within the whole field Engineering Science and Technology, 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.
The current research aimed at fabricating plant extract mediated biosynthesized silver nanoparticles (AgNPs) utilizing thorn extract of Bombax ceiba (TEBC). The synthesized AgNPs was characterized by UV spectroscopy where the surface plasmonic resonance peak (SPR) was located at 222 nm. The scanning electron microscopy (SEM) studies demonstrated that the morphology of fabricated nanomaterials was primarily cylindrical of average size of 20-30 nm with some spindles of size >50 nm. The anti-microbial evaluation against Staphylococcus aureus revealed that AgNPs exhibited notable activity with ZOI of 27.2 mm at MIC of 25 μg/mL. The outcome of this research evidently signified that the biofabricated AgNPs using TEBC may be a new greener approach or technology to formulate anti-bacterial nanodrugs in future.
Comparative study on screening methods of polyhydroxybutyrate (PHB) producing...inventionjournals
International Journal of Engineering and Science Invention (IJESI) is an international journal intended for professionals and researchers in all fields of computer science and electronics. IJESI publishes research articles and reviews within the whole field Engineering Science and Technology, 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.
Inhibitory effect of zinc oxide nanoparticles on pseudomonas aeruginosa biofi...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
Bacterial biofilm formation causes many persistent and chronic infections. The matrix protects biofilm bacteria from exposure to innate immune defenses and antibiotic treatments. The purpose of this study was to evaluate the biofilm formation of clinical isolates of Pseudomonas aeruginosa and the activity of zinc oxide nanoparticles (ZnO NPs) on biofilm.
Materials and Methods:
After collecting bacteria from clinical samples of hospitalized patients, the ability of organisms were evaluated to create biofilm by tissue culture plate (TCP) assay. ZnO NPs were synthesized by sol gel method and the efficacy of different concentrations (50- 350 µg/ml) of ZnO NPs was assessed on biofilm formation and also elimination of pre-formed biofilm by using TCP method.
Results:
The average diameter of synthesized ZnO NPs was 20 nm. The minimum inhibitory concentration of nanoparticles was 150- 158 μg/ml and the minimum bactericidal concentration was higher (325 µg/ml). All 15 clinical isolates of P. aeruginosa were able to produce biofilm. Treating the organisms with nanoparticles at concentrations of 350 μg/ml resulted in more than 94% inhibition in OD reduction%. Molecular analysis showed that the presence of mRNA of pslA gene after treating bacteria with ZnO NPs for 30 minutes.
Conclusion:
The results showed that ZnO NPs can inhibit the establishment of P. aeruginosa biofilms and have less effective in removing pre-formed biofilm. However the tested nanoparticles exhibited anti-biofilm effect, but mRNA of pslA gene could be still detected in the medium by RT-PCR technique after 30 minutes treatment with ZnO.
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.
ABSTRACT In the present study, the extracellular synthesis of Silver nanoparticles was done using two different bacterial strains viz. Bacillus flexus and Bacillus pseudomycoides. The silver nanoparticles were confirmed by in color test and characterized by UV-Visible spectroscopy, the λmax observed at 430 nm and 410 nm confirmed the synthesis of AgNPs. FTIR analysis confirms the presence of elemental silver and reveals the dual function of the biological molecule responsible for the reduction and stabilization of AgNPs in the aqueous medium. The XRD showed that silver nanoparticles produced are crystalline in nature with size ranges from 30 to 70 nm. The SEM shows that produced silver nanoparticles are spherical, Pseudo spherical in shape with traces of agglomeration. Further through investigation of Antibiotic Sensitivity/Resistant pattern expressed that out of eighteen virulent enteric bacterial isolates, three isolates showed MAR index equal to 1, which indicates the presence of multiple drug resistance (MDR). MIC values of AgNPs against MDR isolate E7 and K3 was established to be 80 μg/ml whereas, for isolate Sa1 the MIC value was 70 μg/ml. The synergistic effect of antibiotics in conjugation with biologically synthesized AgNPs encourage the susceptibility amongst the tested bacterial cultures; viz. Salmonella followed by Klebsiella and E. coli.
Key-words: Biosynthesis, Synergistic activity, Antibacterial activity, Silver nanoparticles, Multidrug-resistant (MDR)
The effect of silver nanoparticles on Staphylococcus epidermidis biofilm biom...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
Bacterial biofilm has been considered responsible for many deaths and high health costs worldwide. Their better protection against antibacterial agents compared to free living cells leads to poor treatment efficiency. Nanotechnology is promising approach to combat biofilm infections. The aim of the present study was to eradicate Staphylococcus epidermidis biofilm with silver nanoparticles (SNPs).
Materials and Methods:
SNPs were used at different concentrations (two fold dilutions) and incubation times (24, 48, 72 h). The crystal violet staining and pour plate assays were used to assess biofilm biomass and bacterial viability, respectively. The ability of SNPs on biofilm matrix eradication was assessed through optical density ratio (ODr). Positive control was defined as an ODr =1.0.
Results:
The crystal violet assay indicated that the biofilm matrixes were intact at different concentrations of SNOs and incubation times. There were no significant differences between these parameters (P >0.05). Bacterial enumeration studies revealed that higher concentrations of SNPs were more effective in killing bacteria than lower ones. Although, longer incubation times led to enhancement of anti-biofilm activity of SNPs.
Conclusion:
The anti-biofilm activity of SNPs was concentration- and time-dependent. The results of this study highlighted that SNPs were effective against cell viability; however they were ineffective against biomass.
SYNERGISTIC ANTIMICROBIAL ACTIVITIES OF PHYTOESTROGENS IN CRUDE EXTRACTS OF T...lukeman Joseph Ade shittu
Intensive studies on extracts and biologically active compounds isolated from medicinal plants have doubled in the last decade worldwide. However, as a result of paucity of knowledge and folkloric claim on the effectiveness of sesame leaves in infectious disease treatments, we aimed to determine the synergistic antimicrobial activity of essential oils and lignans present in the crude leaves extracts of Sesame radiatum and Sesame indicum. Ethanolic, methanolic and aqueous extracts of both leaves were studied for their in-vitro synergistic antimicrobial activity against both Gram positive and Gram negative micro-organisms, and Yeast using Agar diffusion method. The GC-MS phytochemical screening of methanolic extract showed that the major compounds in essential oils are of carboxylic acids and phenolic groups especially, the most potent antioxidants known to man like sesamol, sesamolin and sesamin among others. Methanolic and ethanolic extracts have broad spectrum antimicrobial effect against all the tested pathogenic micro-organisms except Streptococcus pneumoniae and Staphylococcus aureus respectively, while the aqueous extract exhibited inhibitory activity on Staphylococcus aureus, Streptococcus pneumoniae and Candida albicans. The result confirmed the folkloric claims of the antimicrobial effectiveness of locally consumed sesame leaves extracts especially against bacterial and common skin infection in many areas of Nigeria .
Effects of combination of magnesium and zinc oxide nanoparticles and heat on ...Nanomedicine Journal (NMJ)
Objective: The objective of this study was to investigate the antibacterial activities of combination of MgO and ZnO nanoparticles in the presence of heat against Escherichia coli and Staphylococcus aureus.
Materials and Methods:Bacteria were grown on either agar or broth media followed by the addition of ZnO and MgO nanoparticles. Then the combined effect of ZnO and MgO nanoparticles was investigated. Furthermore, the media containing nanoparticles were treated with mild heat and their synergistic antibacterial activity was investigated against E. coli and S. aureus in milk.
Results: The data showed that the nanoparticles used in this study had no effect on the bacteria in the agar medium. However, the results showed that ZnO and MgO nanoparticles resulted in a significant decrease in the number of E. coli (P<0.000) and S. aureus (Pd”0.05) in the broth medium. The combination of nanoparticles and mild heat exhibited a significant decrease in the number of E. coli and S. aureus indicating the synergistic effects of nanoparticles and heat.
Conclusion: Using a combination of mild heat, ZnO and MgO nanoparticles, E. coli and S. aureus can be controlled successfully in the milk. Mild heating plus ZnO and MgO nanoparticles has a synergistic effect which would reduce the need for high temperature and also the concentrations of ZnO and MgO nanoparticles required for pathogen control in minimally processed milk during maintaining.
Green synthesis of silver nanoparticles: The reasons for and against Aspergil...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
The enzymatic activity of fungi has recently inspired the scientists with re-explore the fungi as potential biofactories rather than the causing agents of humans and plants infections. In very recent years, fungi are considered as worthy, applicable and available candidates for synthesis of smaller gold, silver and other nano-sized particles.
Materials and Methods:
A standard strain of Aspergillus parasiticus was grown on a liquid medium containing mineral salt. The cell-free filtrate of the culture was then obtained and subjected to synthesize SNPs while expose with 1mM of AgNO3. Further characterization of synthesized SNPs was performed afterward. In addition, antifungal activity of synthesized SNPs was evaluated against a standard strain of Candida albicans. The reduction of Ag+ ions to metal nanoparticles was investigated virtually by tracing the color of the solution which turned into reddish-brown after 72h.
Results:
The UV-vis spectra demonstrated a broad peak centering at 400nm which corresponds to the particle size much less than 70nm. The results of TEM demonstrated that the particles were formed fairly uniform, spherical, and small in size with almost 90% in 5-30nm range. The zeta potential of silver nanoparticles was negative and equal to -15.0 which meets the quality and suggested that there was not much aggression. Silver nanoparticles synthesized by A. parasiticus showed antifungal activity against yeast strain tested and exhibited MIC value of 4 μg/mL.
Conclusion:
The filamentous fungus, A. parasiticus has successfully demonstrated potential for extra cellular synthesis of fairly monodispersed, tiny silver nanoparticles.
Abstract
Objective(s):
In recent years, the biosynthesis of gold nanoparticles has been the focus of interest because of their emerging application in a number of areas such as biomedicine. In the present study we report the extracellular biosynthesis of gold nanoparticles (AuNPs) by using a positive bacterium named Streptomyces fulvissimus isolate U from rice fields of Guilan Province, Iran.
Materials and Methods:
From over 20 Streptomyces isolates collected, isolate U showed high AuNPs biosynthesis activity. To determine its taxonomical identity, its morphology was characterized by scanning electron microscope and partial molecular analysis performed by PCR. In this regard, 16S rDNA of isolate U was amplified using universal bacterial primers FD1 and RP2. The PCR products were purified and sequenced. Sequence analysis of 16S rDNA was then conducted using NCBI BLAST method. In biosynthesis of AuNPs by this bacterium, the biomass of bacterium exposed to the HAuCl4 solution.
Results:
The nanoparticles obtained were characterized by UV-Visible spectroscopy, transmission electron microscopy (TEM) and Energy dispersive X-ray (EDX) spectroscopy and X-ray diffraction spectroscopy (XRD) analyses. Our results indicated that Streptomyces fulvissimus isolateU bio-synthesizes extracellular AuNPs in the range of 20-50 nm.
Conclusions:
This technique of green synthesis of AuNPs by a microbial source may become a promising method because of its environmental safety. Its optimization may make it a potential procedure for industrial production of gold nanoparticles.
— 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.
Genotyping and subgenotyping of Trichophyton rubrum isolated from dermatophyt...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Inhibitory effect of zinc oxide nanoparticles on pseudomonas aeruginosa biofi...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
Bacterial biofilm formation causes many persistent and chronic infections. The matrix protects biofilm bacteria from exposure to innate immune defenses and antibiotic treatments. The purpose of this study was to evaluate the biofilm formation of clinical isolates of Pseudomonas aeruginosa and the activity of zinc oxide nanoparticles (ZnO NPs) on biofilm.
Materials and Methods:
After collecting bacteria from clinical samples of hospitalized patients, the ability of organisms were evaluated to create biofilm by tissue culture plate (TCP) assay. ZnO NPs were synthesized by sol gel method and the efficacy of different concentrations (50- 350 µg/ml) of ZnO NPs was assessed on biofilm formation and also elimination of pre-formed biofilm by using TCP method.
Results:
The average diameter of synthesized ZnO NPs was 20 nm. The minimum inhibitory concentration of nanoparticles was 150- 158 μg/ml and the minimum bactericidal concentration was higher (325 µg/ml). All 15 clinical isolates of P. aeruginosa were able to produce biofilm. Treating the organisms with nanoparticles at concentrations of 350 μg/ml resulted in more than 94% inhibition in OD reduction%. Molecular analysis showed that the presence of mRNA of pslA gene after treating bacteria with ZnO NPs for 30 minutes.
Conclusion:
The results showed that ZnO NPs can inhibit the establishment of P. aeruginosa biofilms and have less effective in removing pre-formed biofilm. However the tested nanoparticles exhibited anti-biofilm effect, but mRNA of pslA gene could be still detected in the medium by RT-PCR technique after 30 minutes treatment with ZnO.
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.
ABSTRACT In the present study, the extracellular synthesis of Silver nanoparticles was done using two different bacterial strains viz. Bacillus flexus and Bacillus pseudomycoides. The silver nanoparticles were confirmed by in color test and characterized by UV-Visible spectroscopy, the λmax observed at 430 nm and 410 nm confirmed the synthesis of AgNPs. FTIR analysis confirms the presence of elemental silver and reveals the dual function of the biological molecule responsible for the reduction and stabilization of AgNPs in the aqueous medium. The XRD showed that silver nanoparticles produced are crystalline in nature with size ranges from 30 to 70 nm. The SEM shows that produced silver nanoparticles are spherical, Pseudo spherical in shape with traces of agglomeration. Further through investigation of Antibiotic Sensitivity/Resistant pattern expressed that out of eighteen virulent enteric bacterial isolates, three isolates showed MAR index equal to 1, which indicates the presence of multiple drug resistance (MDR). MIC values of AgNPs against MDR isolate E7 and K3 was established to be 80 μg/ml whereas, for isolate Sa1 the MIC value was 70 μg/ml. The synergistic effect of antibiotics in conjugation with biologically synthesized AgNPs encourage the susceptibility amongst the tested bacterial cultures; viz. Salmonella followed by Klebsiella and E. coli.
Key-words: Biosynthesis, Synergistic activity, Antibacterial activity, Silver nanoparticles, Multidrug-resistant (MDR)
The effect of silver nanoparticles on Staphylococcus epidermidis biofilm biom...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
Bacterial biofilm has been considered responsible for many deaths and high health costs worldwide. Their better protection against antibacterial agents compared to free living cells leads to poor treatment efficiency. Nanotechnology is promising approach to combat biofilm infections. The aim of the present study was to eradicate Staphylococcus epidermidis biofilm with silver nanoparticles (SNPs).
Materials and Methods:
SNPs were used at different concentrations (two fold dilutions) and incubation times (24, 48, 72 h). The crystal violet staining and pour plate assays were used to assess biofilm biomass and bacterial viability, respectively. The ability of SNPs on biofilm matrix eradication was assessed through optical density ratio (ODr). Positive control was defined as an ODr =1.0.
Results:
The crystal violet assay indicated that the biofilm matrixes were intact at different concentrations of SNOs and incubation times. There were no significant differences between these parameters (P >0.05). Bacterial enumeration studies revealed that higher concentrations of SNPs were more effective in killing bacteria than lower ones. Although, longer incubation times led to enhancement of anti-biofilm activity of SNPs.
Conclusion:
The anti-biofilm activity of SNPs was concentration- and time-dependent. The results of this study highlighted that SNPs were effective against cell viability; however they were ineffective against biomass.
SYNERGISTIC ANTIMICROBIAL ACTIVITIES OF PHYTOESTROGENS IN CRUDE EXTRACTS OF T...lukeman Joseph Ade shittu
Intensive studies on extracts and biologically active compounds isolated from medicinal plants have doubled in the last decade worldwide. However, as a result of paucity of knowledge and folkloric claim on the effectiveness of sesame leaves in infectious disease treatments, we aimed to determine the synergistic antimicrobial activity of essential oils and lignans present in the crude leaves extracts of Sesame radiatum and Sesame indicum. Ethanolic, methanolic and aqueous extracts of both leaves were studied for their in-vitro synergistic antimicrobial activity against both Gram positive and Gram negative micro-organisms, and Yeast using Agar diffusion method. The GC-MS phytochemical screening of methanolic extract showed that the major compounds in essential oils are of carboxylic acids and phenolic groups especially, the most potent antioxidants known to man like sesamol, sesamolin and sesamin among others. Methanolic and ethanolic extracts have broad spectrum antimicrobial effect against all the tested pathogenic micro-organisms except Streptococcus pneumoniae and Staphylococcus aureus respectively, while the aqueous extract exhibited inhibitory activity on Staphylococcus aureus, Streptococcus pneumoniae and Candida albicans. The result confirmed the folkloric claims of the antimicrobial effectiveness of locally consumed sesame leaves extracts especially against bacterial and common skin infection in many areas of Nigeria .
Effects of combination of magnesium and zinc oxide nanoparticles and heat on ...Nanomedicine Journal (NMJ)
Objective: The objective of this study was to investigate the antibacterial activities of combination of MgO and ZnO nanoparticles in the presence of heat against Escherichia coli and Staphylococcus aureus.
Materials and Methods:Bacteria were grown on either agar or broth media followed by the addition of ZnO and MgO nanoparticles. Then the combined effect of ZnO and MgO nanoparticles was investigated. Furthermore, the media containing nanoparticles were treated with mild heat and their synergistic antibacterial activity was investigated against E. coli and S. aureus in milk.
Results: The data showed that the nanoparticles used in this study had no effect on the bacteria in the agar medium. However, the results showed that ZnO and MgO nanoparticles resulted in a significant decrease in the number of E. coli (P<0.000) and S. aureus (Pd”0.05) in the broth medium. The combination of nanoparticles and mild heat exhibited a significant decrease in the number of E. coli and S. aureus indicating the synergistic effects of nanoparticles and heat.
Conclusion: Using a combination of mild heat, ZnO and MgO nanoparticles, E. coli and S. aureus can be controlled successfully in the milk. Mild heating plus ZnO and MgO nanoparticles has a synergistic effect which would reduce the need for high temperature and also the concentrations of ZnO and MgO nanoparticles required for pathogen control in minimally processed milk during maintaining.
Green synthesis of silver nanoparticles: The reasons for and against Aspergil...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
The enzymatic activity of fungi has recently inspired the scientists with re-explore the fungi as potential biofactories rather than the causing agents of humans and plants infections. In very recent years, fungi are considered as worthy, applicable and available candidates for synthesis of smaller gold, silver and other nano-sized particles.
Materials and Methods:
A standard strain of Aspergillus parasiticus was grown on a liquid medium containing mineral salt. The cell-free filtrate of the culture was then obtained and subjected to synthesize SNPs while expose with 1mM of AgNO3. Further characterization of synthesized SNPs was performed afterward. In addition, antifungal activity of synthesized SNPs was evaluated against a standard strain of Candida albicans. The reduction of Ag+ ions to metal nanoparticles was investigated virtually by tracing the color of the solution which turned into reddish-brown after 72h.
Results:
The UV-vis spectra demonstrated a broad peak centering at 400nm which corresponds to the particle size much less than 70nm. The results of TEM demonstrated that the particles were formed fairly uniform, spherical, and small in size with almost 90% in 5-30nm range. The zeta potential of silver nanoparticles was negative and equal to -15.0 which meets the quality and suggested that there was not much aggression. Silver nanoparticles synthesized by A. parasiticus showed antifungal activity against yeast strain tested and exhibited MIC value of 4 μg/mL.
Conclusion:
The filamentous fungus, A. parasiticus has successfully demonstrated potential for extra cellular synthesis of fairly monodispersed, tiny silver nanoparticles.
Abstract
Objective(s):
In recent years, the biosynthesis of gold nanoparticles has been the focus of interest because of their emerging application in a number of areas such as biomedicine. In the present study we report the extracellular biosynthesis of gold nanoparticles (AuNPs) by using a positive bacterium named Streptomyces fulvissimus isolate U from rice fields of Guilan Province, Iran.
Materials and Methods:
From over 20 Streptomyces isolates collected, isolate U showed high AuNPs biosynthesis activity. To determine its taxonomical identity, its morphology was characterized by scanning electron microscope and partial molecular analysis performed by PCR. In this regard, 16S rDNA of isolate U was amplified using universal bacterial primers FD1 and RP2. The PCR products were purified and sequenced. Sequence analysis of 16S rDNA was then conducted using NCBI BLAST method. In biosynthesis of AuNPs by this bacterium, the biomass of bacterium exposed to the HAuCl4 solution.
Results:
The nanoparticles obtained were characterized by UV-Visible spectroscopy, transmission electron microscopy (TEM) and Energy dispersive X-ray (EDX) spectroscopy and X-ray diffraction spectroscopy (XRD) analyses. Our results indicated that Streptomyces fulvissimus isolateU bio-synthesizes extracellular AuNPs in the range of 20-50 nm.
Conclusions:
This technique of green synthesis of AuNPs by a microbial source may become a promising method because of its environmental safety. Its optimization may make it a potential procedure for industrial production of gold nanoparticles.
— 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.
Genotyping and subgenotyping of Trichophyton rubrum isolated from dermatophyt...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
La responsabilità sociale d'impresa e la promozione della salutePina Lalli
Intervento nel Seminario "La promozione della salute nei luoghi di lavoro - Il ruolo del Medico competente" organizzato dal Dipartimento di Sanità Pubblica, Servizio Prevenzione e Sicurezza Ambienti di Lavoro, Azienda Usl di Modena, 24 maggio 2013
Silver Nanoparticles from a Plant Echinacea Purpurea Extract for its Antipath...ijtsrd
Nanotechnology is gaining tremendous impetus in the present century due to its capability of modulating metals into their nanosize. The synthesis, characterization, and application of biologically synthesized nanomaterials have become an important branch of nanotechnology. Research in nanotechnology highlights the possibility of green chemistry pathways to produce technologically important nanomaterials. Metallic nanoparticles are traditionally synthesized by wet chemical techniques, where the chemicals used are often toxic and flammable. Silver nanoparticles are the metal of choice as they hold the promise to kill microbes effectively. The present study describes a cost effective and environment friendly technique for green synthesis of silver nanoparticles from 1mM silver nitrate solution through the a plant Echinacea purpurea cone flower extract. The appearance of brown colour indicates the synthesis of silver nanoparticles. Nanoparticles were characterized using UV Vis absorption spectroscopy and SEM analysis. UV Vis spectrum of the aqueous medium containing silver nanoparticles showed absorption peak at 450nm. SEM analysis showed the average particle size of 50 70nm and spherical shape of the silver nanoparticles. Further studies on the silver nanoparticles showed that it has the antibacterial activities. Antipathogenic activity study was carried out by spread plate, pour plate on Escherichia coli and disc diffusion methods on pathogenic organisms such as Escherichia coli, Proteus vulgaricus, Klebsiella pneumoniae, Pseudomonas aeruginosa. Compared to spread plate, pour plate method showed the maximum antibacterial activity. Zone of inhibition was observed by disc diffusion methods and among these four pathogens, Klebsiella pneumoniae and Escherichia coli showed the maximum activity. Satheesh Kumar B. "Silver Nanoparticles from a Plant Echinacea Purpurea Extract for its Antipathogenic Efficacy" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-6 | Issue-6 , October 2022, URL: https://www.ijtsrd.com/papers/ijtsrd50001.pdf Paper URL: https://www.ijtsrd.com/biological-science/biotechnology/50001/silver-nanoparticles-from-a-plant-echinacea-purpurea-extract-for-its-antipathogenic-efficacy/satheesh-kumar-b
Synthesis and evaluation of bactericidal properties of CuO nanoparticles agai...Nanomedicine Journal (NMJ)
Objective(s):
CuO is one of the most important transition metal oxides due to its captivating properties. It is used in various technological applications such as high critical temperature superconductors, gas sensors, in photoconductive applications, and so on. Recently, it has been used as an antimicrobial agent against various bacterial species.
Materials and Methods:
Here, we synthesized CuO nanoparticles (NPs) and explored the antibacterial activity of CuO NPs preparation.
Results:
Single crystalline nanoparticles of copper oxide having almost uniform particle size of 5-6 nm has been synthesized by a facile and versatile route. XRD spectra confirmed the formation of single phase CuO NPs. Transmission electron microscopy results corroborate well with XRD results. The technique employed is free from toxic solvents, organics and amines, is based on a simple reaction of copper sulfate and de-ionized water (DI), and their bactericidal effects against of Aeromonas hydrophila ATCC 7966T bacteria were investigated. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) with liquid culture for all of the Aeromonas hydrophila culture Medias was done.
Conclusion:
Present study confirms that Copper oxide nanoparticles have great promise as antimicrobial agent against Aeromonas hydrophila.
In recent years, nanoparticles that have size of 1-100 nm is widely used for textile, pharmacy,
cosmetic and treatment of industrial wastewater. Producing and using of nanoparticles widely, causes
important accumulation in nature and toxicity on ecosystem. Knowledge of potential toxicity of nanoparticles is
limited. In this study, six different nanoparticles nano-zinc oxide, nano-silicon dioxide, nano-cerium oxide,
nano-aluminum oxide, nano-hafnium oxide, and nano-tantalum oxide which used commonly, were studied to
investigate toxic impacts on organisms. We studied nine different acute toxicity test (bacteria – Escherichia coli
(gram negative bacteria) ; bacteria – Bacillus cereus (gram positive bacteria) ; bacteria – Vibrio fischeri
(bioluminescences bacteria) ; methane Archae Bacteria ; yeast – Candida albicans ; mold – Aspergillus niger ;
algae – Chlorella sp. ; Crustacea – Daphnia magna ; lepistes - Poecillia reticula) for the effect of
nanoparticles to different trophic levels. In general, the most toxic nanoparticle is nano-zinc oxide and the least
toxic nanoparticle is nano-hafnium oxide. Among the used organisms in acute toxicity test; the most sensitive
organism is algae - Chlorella sp ;the most resistant organism is fish- Poecillia reticula.
ABSTRACT- In our present study, we account for eco-friendly biosynthesis of copper nanoparticles using aqueous leaves extract of Artocarpus heterophyllus against first to fourth instar larvae of Aedes aegypti. The synthesized CuNPs were characterized by UV, XRD, FTIR and SEM analyses were clearly distinguishable. The four different immature mosquito larval stages of A. aegypti were exposed to varying concentrations of aqueous leaf extract of A. heterophyllus, copper sulphate (CuSO4) and synthesized CuNPs for 24 h. The mortality was observed at aqueous extract (LC50= 48.40, 60.55, 70.36, and 82.79 mg/ml), CuSO4 (LC50=21.81, 26.92, 41.38, and 55.12 mg/ml) synthesized CuNPs against first to fourth instars of A. aegypti (LC50= 3.85, 4.24, 4.66, and 5.08 mg/ml), respectively. The novel properties created not only improve the quality of human’s life; it also helps in saving energy and environment.
Key-words: Copper nanoparticles, Jackfruit, Aedes aegypti, aqueous leaf extract
Green synthesis of silver nanoparticles: The reasons for and against Aspergil...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
The enzymatic activity of fungi has recently inspired the scientists with re-explore the fungi as potential biofactories rather than the causing agents of humans and plants infections. In very recent years, fungi are considered as worthy, applicable and available candidates for synthesis of smaller gold, silver and other nano-sized particles.
Materials and Methods:
A standard strain of Aspergillus parasiticus was grown on a liquid medium containing mineral salt. The cell-free filtrate of the culture was then obtained and subjected to synthesize SNPs while expose with 1mM of AgNO3. Further characterization of synthesized SNPs was performed afterward. In addition, antifungal activity of synthesized SNPs was evaluated against a standard strain of Candida albicans. The reduction of Ag+ ions to metal nanoparticles was investigated virtually by tracing the color of the solution which turned into reddish-brown after 72h.
Results:
The UV-vis spectra demonstrated a broad peak centering at 400nm which corresponds to the particle size much less than 70nm. The results of TEM demonstrated that the particles were formed fairly uniform, spherical, and small in size with almost 90% in 5-30nm range. The zeta potential of silver nanoparticles was negative and equal to -15.0 which meets the quality and suggested that there was not much aggression. Silver nanoparticles synthesized by A. parasiticus showed antifungal activity against yeast strain tested and exhibited MIC value of 4 μg/mL.
Conclusion:
The filamentous fungus, A. parasiticus has successfully demonstrated potential for extra cellular synthesis of fairly monodispersed, tiny silver nanoparticles.
Nanobiotechnology, bionanotechnology, and nanobiology are terms that refer to the intersection of nanotechnology and biology. Given that the subject is one that has only emerged very recently, bionanotechnology and nanobiotechnology serve as blanket terms for various related technologies.
Enlargement of biologically stimulated
investigational processes for the synthesis of nanoparticles is
budding into an important branch of nanotechnology. Eco
responsive methods of green mediated synthesis of nanoparticles
are the present research in the extremity of nanotechnology. The
bioreduction behavior of leaf extracts of Morinda citrifolia L.
(Rubiaceae) in the green synthesis of silver nanoparticles was
investigated employing UV/Visible Spectrophotometry, Particle
size analyzer, Zeta potential, Filed emission scanning electron
microscopy, Energy Dispersive X-ray Analysis and FourierTransform
Infrared Spectroscopy. The antifungal property of the
silver nanoparticles was tested against Candida albicans,
Candida tropicalis and Candida krusei. The Antifungal assay
tests Zone of inhibition revealed the concentrations of more than
10µl of silver nanoparticles were inhibited the growth of fungal
pathogens.
ABSTRACT- In this study, the effect of ZnO and TiO2-NPs on beneficial soil microorganisms and their secondary metabolites production was investigated. The antibacterial potential of NPs were determined by growth kinetics of P. aeruginosa, P. fluorescens and B. amyloliquefaciens. Significantly decreased in the cell viability based on optical density measurements were observed upon treatment with increasing concentrations of NPs. While comparing the effect of the different concentrations of the NPs (200 µg/ml) on IAA production by different bacterial strains, ZnO nanoparticles showed greater inhibitory effect than TiO2-NPs on IAA production by bacterial strains. The effect of Nanoparticles on phosphate solubilization was found inhibitory at 200 µg/ml. Treatment with ZnO showed concentration dependent enhancement in siderophore production by bacteriaby exposure to ZnO-NPs whereas TiO2-NPs showed concentration dependent progressive decline for iron binding siderophore molecules. Reduction in antibiotic production by P. aeruginosa and P. fluorescens was noticed in the presence of ZnO and TiO2 as compared to the control. The fluorescence of NADH released by P. aeruginosa was observed to be quenched in presence of ZnO and TiO2-NPs as compared to control. The present study highlights that the impact of nanoparticles on bacterial strains and the release of plant growth promoting substances by PGPR strains was dose dependent, which gives an idea about the level of toxicity of these nanoparticles in the environment. Therefore, the discharge of nanoparticles in the environment should be carefully monitored so that the loss of both structure and functions of agronomically important microbes could be protected from the toxicity of MO-NPs.
Key-words- MO-NPs, IAA, Phosphate Solubilization, Siderophore, PCA, NADH, ZnO-NPs, TiO2-NPs
Recently, biosynthesis of metal nanoparticles has
drawn considerable attention due to environment friendly and
sustainable methods. Herein, Bacillus subtilis TD 6 was selected
as candidate for biosynthesis of silver nanoparticles (AgNPs).
These nano-sized silver particles (AgNPs) at concentration of 500
ppm were synthesized using maize and potato starch as both
reducing and stabilizing agent. A solution containing AgNPs (500
ppm) was diluted with distilled water to 100 and 50 ppm and
applied to cotton fabrics in presence of binder. The finished
fabric characterization was carried out using UV-Vis
spectroscopy. The antibacterial activity of the treated fabrics
loaded with AgNPs was evaluated against Escherichia coli (gram
negative) and Staphylococcus aureus (gram positive) bacteria.
Results explored that, binder retains excellent antibacterial
properties even after 20 washing cycles reflecting the significance
of binder in fixation of AgNPs deposits on the surface of the
fabrics.
Cytotoxic | Primary research | Silver nanoparticlesPubrica
Biosynthesis, Antimicrobial, and Cytotoxic Effects of Silver Nanoparticles Using Acacia Concinna POD Extract and Kigelia Africa Leaf Extract- Secondary metabolites found in plants include alkaloids, flavonoids, phenolic compounds, phytosterols, saponins, tannins, carbohydrates, proteins, lipids, and minerals. These secondary metabolites have a wide range of uses, including the production of nanoparticles.
Biosynthesis, Antimicrobial, and Cytotoxic Effects of Silver Nanoparticles Using Acacia Concinna POD Extract and Kigelia Africa Leaf Extract- Secondary metabolites found in plants include alkaloids, flavonoids, phenolic compounds, phytosterols, saponins, tannins, carbohydrates, proteins, lipids, and minerals. These secondary metabolites have a wide range of uses, including the production of nanoparticles.
Visit us @ https://pubrica.com/insights/sample-work/cytotoxic-effects-of-sliver-nanoparticles-using-leaf-extract/
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
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1. Introduction
Fungi are good natural source for production of many bioactive agents that can be used in medical
and pharmaceutical applications (Lindequist et al., 2005; Demain and Sanchez, 2009). Nevertheless,
the last decades have witnessed an increased research interests in nanobiotechnology to explore
natural sources for biosynthesis of metal nanoparticles as agents widely used in biomedical aspects
(Sharma et al., 2008; Marambio-Jones and Hoek, 2012; Kashyap et al., 2013). So far noteworthy
studies have been conducted on the use of fungi as bionanofactories for synthesis of metal
nanoparticles that can be implemented in drug industry and medical therapy (Feng et al. 2000;
Aymonier et al., 2002; Sastry et al., 2003; Rai et al., 2009). Silver nanoparticles are one the most
significant nanoametals exhibit a high potentiality as antimicrobial agent against pathogenic
bacteria and fungi and a number of fungal species so far have been explored for their capability for
synthesis of silver nanoparticles (Morones et al., 2005; Lee et al., 2006; Riddin et al., 2006; Kim et
al., 2007; Yokoyama and Welchons 2007; Ingle et al. 2008; Maliszewska et al., 2009; Fayaz et al.,
2010; Gade et al., 2010).
Nowadays, however, more attention has been given to explore potent biosynthesized nanoparticles
from fungi using nanobiotechnology approach for cancer therapy (Amiji 2007; Verma et al., 2013).
Nonetheless, biosynthesis of silver nanoparticles by fungi is of significance due to the fungal
characteristics such as easy to culture giving large biomass productivity, secretion of extracellular
enzymes and eco-friendly (Sastry et al., 2003; Rai et al., 2009; Kashyap et al., 2013). The present
work aimed to explore for the first time the endophytic fungus Papulaspora pallidula for its
capability to synthesize and characterize silver nanoparticles and examine their efficacy as
antitumor against human larynx carcinoma cell line (Hep-2), their activity against five strains of
human pathogenic bacteria and their efficacy as antibacterial in a combination with the commercial
antibiotic Gentamycin.
2. Materials and Methods
2.1 Isolation of the endophytic fungus
The endophyte fungus Papulaspora pallidula was isolated from plant roots of Mesembryanthemum
sp.(Family Aizoaceae) collected from natural habitats in Basra (Southern Iraq) during the year
2014. The plant roots were cut in to small segments (5 cm long) and rinsed in tap water for 10 min
followed by rinse in sterile distilled water. Root segment surfaces were sterilized with 70% ethanol
(for 5 min) followed by 5% sodium hypochlorite solution for 2 min then rinsed in 90% ethanol for
1 min and kept in 10% NaHCO3 to reduce the growth of any fungal species associated with the
plant roots surfaces. The root segments were rinsed three times with sterilized distilled water,
dried on sterilized filter paper and placed onto potato dextrose agar plates (PDA) supplemented
with 20 μg/ml of tetracycline to restrict bacterial growth (Verma at el. 2009). The plates were
incubated at 25 °
C for 14 days, examined for any growth of endophytic fungi and pure cultures of
the growing fungi on PDA were made. The isolated endophytic fungus was identified following the
taxonomic description (Ellis, 1971; Domsch et al., 1980; Watanabe, 2002).
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2.2 Fungal culture conditions and biosynthesis of Silver Nanoparticles
The isolated endophytic fungus was grown in a liquid medium of potato dextrose broth (PDB) (250
g potato and 20 g dextrose per liter of distilled water) in 250 ml flasks adjusted at pH 5.6 and
incubated at 27 °C in a static condition. After 10 days of incubation the mycelia biomass was
separated by filtration and washed thoroughly with sterilized Milli-Q deionized water. For
extraction of the biosynthesized AgNPs, 10 g of the fungal biomass (wet weight) was brought in
contact with 100 ml sterile distilled water in Erlenmeyer flasks, incubated at 27°C in a rotary
shaker at 120 rpm. After incubation the fungal free-cell filtrate was obtained by filtration using
Whatman filter paper No 1. 100 mL of fungal free-cell filtrate (fcf) was amended with 0.017 g of
silver nitrate AgNO3 (Alfa Aesar 99.9%, Germany) to get a final concentration of 1 mM and
incubated at 27°C under dark condition (Ingle et al., 2009).
2.3 Detection and characterization of silver nanoparticles
The biosynthesized silver nanoparticles (AgNPs) within the fungal free-cell filtrate was visually
examined by changing of color from pale yellow to dark brown and was further confirmed by UV-
Vis spectrophotometer.
2.4 UV-Visible spectrophotometric assay
After 24 hr of incubation of the fungal free-cell filtrate treated with AgNO3, the bioreduction of Ag+
in aqueous solution was monitored using UV-Vis spectrophotometer (APEL PD-303, Japan) at
regular intervals. During the reduction process, 0.1ml of filtrate was taken and diluted three times
with deionized water, centrifuged at 800 rpm for 5 min and the supernatant was scanned using UV-
Vis spectrophotometer at the wave lengths of 300 to 900 nm. UV-Vis spectra were recorded at 24,
48, and 72 hr at a resolution of 1 nm. Untreated free- cell filtrate was used as a control.
2.5 Fourier Transform Infrared (FTIR)
After 72 hr of incubation, the free-cell filtrate was subjected to Fourier Transform Infrared (FTIR)
(Shimadzu UV-1700, Japan) analysis. After a complete reduction of silver ions within the fungal
filtrate, the filtrate was mixed with acetone (1:5 vol /vol) with a continuous shaking then
centrifuged at 4000 rpm for 15 min forming a pellet. The supernatant was discarded and 2 ml of
acetone was added into the pellet and shaken thoroughly then poured into a Petri plate and the
acetone was evaporated in order to obtain powder of silver nanoparticles. Characterization of
AgNPs was carried out by using FTIR at the range of 400– 4000 cm-1at a resolution of 4 cm-1
(Raheman et al., 2011).
2.6 SEM analysis
Scanning Electron Microscopic (SEM) (Netherland INSPECT S50) analysis of the fungal free-cell
filtrate treated with AgNO3 was performed. A sample from the free- cell filtrate was filtered
through Millipore filter of 0.2 μm pore size to remove any contaminations interfering with the SEM
images. Thin films of the filtrate were prepared on a carbon coated copper grid by just dropping a
very little amount of the filtrate on the grid and the extra solution was removed by a blotting paper
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then the films on the grids were allowed to dry overnight at room temperature under a sterilized
condition. SEM micrographs of the silver nanoparticles were exposed at different magnifications.
2.7 Antitumor efficacy of silver nanoparticles assay
In vitro the cytotoxicity effect of the biosynthesized silver nanoparticles against human larynx
carcinoma cell line (Hep-2) was tested. Human larynx carcinoma cell line Hep-2 was cultured on
specific medium supplemented with antibiotics and the cells were grown under humidity and CO2
conditions at 37 °C. Cell lines (Hep-2) in an amount of (1x105 cell/well) were seeded into 96 well
tissue culture plates (Verma et al., 2013). Different concentrations (0.05, 0.1, 0.2, 0.39, 0.78, 1.56,
3.13, 6.25, 25, and 100 ug/ml) of biosynthesized AgNPs were prepared and amended into the wells
containing the cell lines and incubated at 37 C for 24 hr. Untreated cells were made as control. After
incubation, cells were washed with PBS and the cells viability was determined by the MTT
technique. The absorbance was measured at 550 nm using spectrophotometer (APEL PD-303,
Japan) and the cell viability percentage was calculated as following: Cell viability (%) = Sample
absorbance / Control absorbance ×100. The concentration of silver nanoparticles which kill 50% of
cells (IC50) was determined.
2.8 Antibacterial activity of silver nanoparticles
The potentiality of silver nanoparticles was examined for their antibacterial efficiency using agar
well diffusion assay method (Perez et al., 1990). Five strains of pathogenic bacteria viz. Escherichia
coli, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella typhi and Staphylococcus aureus were
tested. Swabs from each bacterial culture grown overnight were streaked on sterilized Muller-
Hinton agar (MHA) plates. Wells (5 mm diam) were made in agar plates using sterilized stainless
steel Cork borer. The wells were loaded with two concentrations (50 and 100 ul) of silver
nanoparticles solutions, incubated at 37 oC for 24 hr and examined for the appearance of inhibition
zones around the wells and the diameters of inhibition zones were measured.
2.9 Minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) assay
Minimum inhibitory concentration (MIC) assay was carried out using the micro dilution method
according to San and Don (2013). 100μL of AgNPs was transferred into 96-well microtitre plates
containing 100μL of Mueller-Hinton broth. 100μL of the tested bacteria E. coli (ATCC 25922) and S.
aureus (NCTC 6571) was inoculated into each well and incubated at 37 oC for 24 hr. After the
incubation period a small amount of bacterial suspension was streaked on MHA plates and
incubated at the same conditions. The minimum inhibitory concentration (MIC) was determined as
the lowest concentration of AgNPs that inhibits the growth of bacteria. The minimal bactericidal
concentration (MBC) was recorded as the minimum concentration of silver nanoparticles that kill
bacteria and no any visible growth of tested bacteria was observed (Qi et al., 2004).
2.10 Assay of the combined effect of AgNPs with Gentamycin
Disc diffusion method was used to assay the combined effect of synthesized AgNPs with commonly
used antibiotic Gentamycin (Devi and Joshi, 2012). A standard antibiotic disc of Gentamycin was
impregnated with 20 μL of freshly prepared AgNPs and placed onto the MHA medium inoculated
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with each tested bacteria. Standard antibiotic discs were used as positive control and AgNPs discs
were used as negative control. The plates were incubated at 37 oC for 24-48 hr. After incubation, the
zones of growth inhibition for the combination treatment and control plates were measured.
2.11 Assessment of increase in fold area
The increase in fold area was assessed by calculating the mean surface area of the inhibition zone
exhibited by the antibiotic alone and in a combination with AgNPs. The fold increase area was
calculated by the equation; (B2 − A2) / A2, where A refers to the inhibition zone diam exhibited by
the Gentamycin activity alone and B refers to the inhibition zone diam exhibited by activity of a
combination of Gentamycin and AgNPs (Birla et al., 2009).
3. Results
3.1 Endophytic fungal culture and biosynthesis of silver nanoparticles
The fungal free-cell filtrate (fcf) obtained from the fungal cultures grown in PDB liquid medium for
ten days after being treated with AgNO3 turned from pale yellow into dark brown color which
indicates the formation of silver nanoparticles (Fig.1).
Fig. 1. Mycofabrication of silver nanoparticles (AgNPs) by the endophytic fungus Papulaspora
pallidula isolated from the plant Mesembryanthemum sp. after being treated with AgNO3
solution indicated by the color change from pale yellow into dark brown
3.2 UV-Vis spectra of AgNPs
The UV-Vis spectra recorded from the fungal free-cell filtrates amended with 1 mM AgNO3 solution
revealed significant variations in spectra of silver nanoparticles synthesis at different intervals of
reaction (Fig. 2). The absorbance pattern of the AgNPs monitored at the range of 300-900 nm
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revealed an increase of absorbance with increasing time of incubation at 430 nm. Highest spectrum
of AgNPs synthesis after 72 hr of incubation was detected.
Fig. 2. UV-Vis spectrum of fungal free cell filtrate (fcf) containing biosynthesize silver nanoparticles
recorded at different exposure times.
3.3 SEM analysis
SEM images with different magnifications showed that the silver nanoparticles are dispersed or
aggregated and mostly showed spherical shape and their size ranging between 8-90 nm (Fig.3).
Fig. 3. SEM micrograph showing the biosynthesized silver nanoparticles in fungal free-cell filtrate
appeared as spherical shape dispersed or aggregated with size range between 8-90 nm
(magnification X 13000).
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3.4 FTIR spectroscopy
FTIR analysis of silver nanoparticles synthesized from the fungus P. pallidula showed a presence of
peaks at 3411.84 cm-1 and 3448.45 cm-1 (Fig.4) refers to the bonding vibrations of the amide of
proteins while the band at 1409. 87 cm-1 and1471.59 cm-1 indicates the presence of C-N stretching
vibrations of aromatic amines.
Fig. 4. FTIR analysis of biosynthesized silver nanoparticles in fungal free-cell before (a) and after
(b) of silver bioreduction
3.5 Antitumor efficacy of biosynthesized silver nanoparticles
The cytotoxicity in vitro of biosynthesized silver nanoparticles at different concentrations against
human larynx carcinoma cell line (Hep-2) was examined. The results revealed that the inhibition
concentration (IC50) of biosynthesized silver nanoparticles to produce 50% of tumor cells
mortality was at concentration 3.13 ug/ul (Fig. 5). However, the cytotoxicity was increased at
higher concentrations of silver nanoparticles.
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Fig. 5. In vitro the cytotoxicity effect of biosynthesized silver nanoparticles by the fungus P.
pallidula against Human larynx carcinoma cell-line (Hep-2)
3.6 Antibacterial activity of silver nanoparticles
The mycofabricated silver nanoparticles exhibited an antibacterial activity against the tested
strains of Gram positive and Gram negative bacterial strains. The results showed that the bacterial
growth inhibition at 50 ul/ml concentration of AgNPs was slightly lower (12-22 mm inhibition
zones dim) than at 100 ul/ ml (15-24 mm inhibition zones diam) (Fig.6). The highest growth
inhibitory activity of AgNPs was against P. aeruginosa and lowest against P. mirabilis. The minimal
inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values were very low
against the two strains of bacteria E. coli and S. aureus (Table 1).
Fig. 6. The growth inhibition zones exhibited by two concentrations of silver nanoparticles
synthesized by the fungus P. pallidula against five strains of human pathogenic bacteria at
two concentrations of AgNPs
0
5
10
15
20
25
14
15
20
21
22
24
14
16
12
15
Zoneofinhibition
AgNPs 50 μl AgNPs 100 μl
E. coli
S . aureus
P .aeruginosa
S. typhi
P. mirabilis
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Table 1 The minimal inhibitory concentrations (MIC) and minimal bactericidal concentrations
(MBC) of biosynthesized AgNPs against two strains of bacteria.
3.7 Combination efficacy of AgNPs with Gentamycin
The activity of AgNPs combined with commercial antibiotic Gentamycin was significantly increased
against the growth of the tested strains of bacteria compared with activity of either of them alone
(Fig. 7). Over all the tested bacteria, the growth inhibition zones ranged between 27-35.5 mm diam
(Table 2). Among the examined bacteria, the growth of S. typhi, P. mirabilis and E. coli was
remarkably inhibited by silver nanoparticles combined with Gentamycin as indicated by the
increase in fold area values (Table 2).
Table 2 Growth inhibition zones exhibited by the silver nanoparticles combined with commercial
antibiotic gentamycin against five bacteria strains and the calculated increase in fold area
Bacteria
Inhibition Zone (mm diam)
Increase in fold areaAgNPs Gentamycin Ag+Ge
E. coli 18* 22.5 27 0.44**
S. aureus 18 29.5 34.5 0.397**
P. aeruginosa 20 25.5 27 0.12
S. typhi 14.5 25 30.5 0.488**
P. mirabilis 20 30 35.5 0.40**
*Values represent means of three replicates
**Significant differences at P < 0.001
Bacteria MIC(μg/ml) MBC(μg/ml)
E. coli 0.078 0.156
S. aureus 0.019 0.039
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Fig. 7. Antibacterial activity indicated by the inhibition zones (mm diam) exhibited by
biosynthesized silver nanoparticles against E. coli (A), S. aureus (B), P. aeruginosa (C), S.
typhi (D) and P. mirabilis (E) using AgNPs alone (1), commercial antibiotic Gentamycin
(2) and a combination of AgNPs with Gentamycin (3)
4. Discussion
Fungi are a good tool used in nanobiotechnology and recently widely applied in multidisciplinary
fields including medical therapy (Karbasian et al., 2008; Maliszewska et al., 2009). Nonetheless,
during the last decade the researches have been focused on the biosynthesis of metal nanoparticles
by fungi as natural sources and as bionanofactories (Karbasian et al., 2008; Sadawiski et al., 2008).
Among the metals, silver nanoparticles are of importance mainly in medical therapy applications
and so far a number of investigations were carried out on fungi to synthesize silver nanoparticles as
antimicrobial agent (Fayaz et al., 2010; Kashyap et al., 2013; Maliszewska et al., 2009; Mukherjee et
al., 2013; Qi et al., 2004). Despite of the mechanistic that involved in fabrication of silver
nanoparticles by fungi are remain unclear, however, it has been proposed that silver nanoparticles
biosynthesis is related to the enzyme reductase which is responsible for the reduction of Ag+ ions
and synthesis of AgNPs (Duran et al., 2005). Furthermore, a reduction of Ag+ may be due to a
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conjugation between the electrons shuttle with reductase enzyme involvement (Maliszewska et al.,
2009). The present study showed that the selected fungus P. pallidula exhibited a high potentiality
for synthesis of silver nanoparticles in culture medium as indicated by the color change from yellow
into dark brown after 72 h of incubation after being treated with 1 mM AgNO3 solution. These
findings are in concomitant with other previous studies using different fungal species (Ahmad et al.
2003; Birla et al. 2009; Raheman et al., 2011; Verma et al., 2013). The color change after
amendment of AgNO3 in to the fungal free-cell filtrate is due to the excitation of surface plasmon
resonance vibration of silver that confirmed the reduction of silver ions as reported by Chitra and
Annadurai (2013). The present study revealed that UV-Vis spectrophotometry analysis showed a
peak with high absorbance at 430 nm which verified the AgNPs synthesis by the examined fungus
and a completed biosynthesis of AgNPs was after 72 h of free-cell filtrate incubation. This is in
agreement with some other works (Chitra and Annadurai, 2013; Henglein, 1993). In comparison
with other studies it appeared that there are some variations in characterization of silver
nanoparticles biosynthesized by different fungal species (Birla et al. 2009; Chitra and Annadurai,
2013; Maliszewska et al., 2009; Raheman et al., 2011). These variations might be due to the source
of fungal isolates or strains and culture conditions (Marambio-Jones and Hoek, 2012). SEM images
demonstrated that the shape of biosynthesized AgNPs by the selected fungus were mostly spherical
and dispersed with size of 8-90 nm. A recent study (Muhsin and Hachim, 2014) showed that the
size of silver nanoparticles synthesized by the fungus Nigrospora sphaerica was ranged between
20-70 nm while those synthesized by the fungus Curvularia tuberculata was ranged between 10-50
um (Muhsin and Hachim, 2015). Other studies, however, reported a variable shape and size of
silver nanoparticles synthesized by different fungal species (Marambio-Jones and Hoek, 2010;
Martinez-Castanon et al., 2008; Pal et al., 2007). It has been stated that the absorption spectrum of
spherical shape of silver nanoparticles present a maximum between 420-450 nm (Matrinez-
Castanon et al., 2008). Analysis of FTIR indicated the release of proteins into fungal filtrate which
causes a reduction of silver ions present in the free-cell filtrate. The reduction of the Ag+ ions can be
attributed to the enzyme reductase that produced by the fungal hyphae as reported by Gole et al.
(2001). It has been speculated that FTIR analysis has indicated that peptides are binding with the
silver nanoparticles forming a capping agent of nanoparticles and stabilizing them in the fungal
culture medium (Kim et al., 2007; Shahverdi et al., 2007).
Recently, more researches interests has been focused on the biosynthesized silver nanoparticles
as anticancer agents since cancer is a serious disease for human leading to a high mortality over the
world and chemotherapy is the most common mode of cancer treatment (Guranathan et al., 2013;
El Kassas and Attia, 2014). Nevertheless, recent approach using nanobiotechnology to explore new
agents as antitumor from fungi has been introduced (Amiji, 2006; Kim et al., 2013; Verma et al.,
2013). In the present work, the biosynthesized silver nanoparticles by the fungus P. palludula
exhibited a high rate of tumor cells growth inhibition at a concentration (IC50) of 3.31 ug/ul
reaching 50% cell mortality. A study of Verma et al. (2013) showed that the IC50 was at 30 ug/ml
concentration of silver nanoparticles synthesized by Penicillium sp. to produce 50% of tumor cells
mortality. Other investigation (El-Sonbaty, 2013) reported that the LD50 of silver nanoparticles
synthesized by Agaricus bisporus against breast cancer was 50 ug/ml reaching 50% of cancer cells
mortality. Previous studies (Bahimba et al., 2012; Guranathan et al., 2013) suggested that
biosynthesized AgNPs as antitumor agent decrease the development of tumor cells and their
toxicity effect is related to the size of silver nanoparticles. It was also stated that silver
nanoparticles have an impact on the cell membrane integrity and consequently tumor cells death
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(Guranathan et al., 2013). Kim et al. (2013) reported that biosynthesized silver nanoparticles
promote the apoptotic pathway leading into antitumor effect. Nevertheless, in the present work a
dose dependent method was employed to estimate the toxicity of mycosynthesized AgNPs by the
fungus P. pallidula and revealed a high efficacy against human larynx carcinoma cell line (Hep-2) at
a minimal concentration (3.31 ug/ul). Other studies (Guranathan et al., 2013; Kim et al., 2013;
Verma et al., 2013) using different sources of tumor cell-lines reported variable minimal
concentrations of silver nanoparticles to produce 50% tumor cells mortality.
The present investigation also showed that the biosynthesized silver nanoparticles exhibited a
significant growth inhibition at two concentrations (50 μl /ml and 100 μl /ml) against the tested
human pathogenic bacterial strains. However, AgNPs exhibited variations among their antibacterial
efficacy. Although, the mechanism of AgNPs impact on the bacterial growth is not well documented
and it can be related to the effect of Ag ions by causing bacterial cell membrane destruction, enzyme
damage or DNA denaturation as stated by other studies (Kim et al. 2007; Marambio-Jones and
Hoek, 2010). The present data revealed that MIC values were very low indicating that the
biosynthesized silver nanoparticles by the fungus P. palldula exhibits a high antibacterial efficacy
against Gram positive and Gram negative bacteria. Similar findings were reported by other
researchers using AgNPs synthesized from other sources of fungal species (Maliszewska et al.,
2009; Muhsin and Hachim, 2015; Verma et al., 2013). The mycofabricated silver nanoparticles
combined with the antibiotic Gentamycin revealed a significant increased efficacy against the
selected human pathogenic bacteria as expressed by increased fold area according to Birla et al.
(2009). These results support some other studies examined the synergistic effects of AgNPs
synthesized from various fungal species in a combination with different commercial antibiotics
tested against Gram positive and Gram negative bacteria ( Fayaz et al., 2010; Gajbhiye et al., 2009;
Shahverdi et al., 2007).
5. Conclusion
A conclusion can be derived that for the first time the selected endophytic fungus revealed a high
potentiality for synthesis of silver nanoparticles which exhibiting a high growth inhibition rate
against a human larynx carcinoma cell-line (Hep-2) and also showed a wide spectra efficacy against
Gram positive and negative human pathogenic bacteria. This fungus is a promising as a natural
source for synthesis of silver nanoparticles that can be implicated in medical cancer therapy and
pharmaceutical drug industry.
Acknowledgment
We are thankful for the authorities of Basra University (Iraq) for supporting this research work as a
part of MSc. research program scholarship awarded to the second author.
Conflict of Interest
No conflict of interest
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