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Home > Mycosynthesis of Silver Nanoparticles: Optimization, Characterization and Their Antimicrobial and Cytotoxic Activity

Mycosynthesis of Silver Nanoparticles: Optimization, Characterization and Their Antimicrobial and Cytotoxic Activity

Thesis Info

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Author

Shahzad, Asma

Program

PhD

Institute

Lahore College for Women University

City

Lahore

Province

Punjab

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Biotechnology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/10331/1/Asma_Shahzad_Biotech_2019_LCWU_30.04.2019.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726741609

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Silver nanoparticles (AgNPs) have been synthesized in the past by varying physical and chemical approaches that were neither ecologically safe or economical besides being time consuming. Therefore silver nanoparticles (extracellular) were devised by employing fungi taking all the issues into account. Henceforward, in the present study Aspergillus fumigatus BTCB10 (KY486782) was opted for the mycosynthesis of AgNPs after extensive screening. Silver nanoparticles were further optimized by using a classical method of one factor at a time methodology along with more recent approach Research Surface Methodology (RSM) through central composite design (CCD). Various variables were investigated for producing efficient and size controlled silver nanoparticles comprising of temperature (ºC), light/dark conditions, substrate concentration (mM), biomass weight (g), fungal culture age (days), pH, ratio of cell free filtrate (CFF)/silver nitrate, metal salts (%), surfactants and enhancer, nitrogen sources, carbon sources and enzymes. The most effective parameters were then selected for RSM study which contained concentration of silver nitrate (mM), NaCl (%) and the wet weight of biomass (g). According to the results of one factor design a very stable (-23.4 mV) small size of 0.681 nm AgNPs were obtained in the presence of these conditions i.e. incubation in dark, temperature 25 ºC, AgNO3 concentration 1mM, biomass 7g, fungal culture age 7 days, pH 6, ratio of cell free filtrate (CFF)/silver nitrate (3:2), NaCl 20%. UV-Vis band showed presence of peak at 400 nm and Fourier Transform Infrared (ATR-FTIR) confirmed presence of main functional groups (O-H, C=O). Moreover, Scanning Electron Microscopy (SEM) revealed cubeshaped AgNPs and X-Ray Diffraction (XRD) verified crystalline nature of AgNPs. The RSM studies also reduced the size of silver nanoparticles to 33.23 nm (spherical) with negative zeta potential of -9.91 mV, excitation peak at 420 nm and comprised of wide range of functional groups (O-H, C=C, C≡C, C-Br, C-Cl). A significant F- value (3.91), nonsignificant lack of fit and determination coefficient (R2=0.7786) were also calculated within the study. A tremendous size reduction was observed by one factor design (99.79 %) and RSM (78.7%) compared to nanoparticles produced prior to optimization having an average size of 356 nm. High nitrate reductase activity with value of 179.15 nmol/h/ml was attained; thus indicating a possible relation between enzyme production and AgNPs synthesis. Additionally, antibacterial activity was also monitored with varying concentrations 0.1-7 μg/mL of AgNPs against Pseudomonas aeruginosa BTCB01, Staphylococcus aureus BTCB 02, Escherichia coli BTCB03, Klebsiella pneumoniae BTCB04 and Acientobacter BTCB05. Maximum zone of inhibition was observed with Klebsiella pneumoniae BTCB04 (33mm) at 7 μg/mL and minimum with Pseudomonas aeruginosa BTCB01 and Staphylococcus aureus BTCB 02 (11mm) at 0.1 μg/mL. Combinatorial effect with streptomycin (1 μgmL-1) was also studied with all the strains, Klebsiella pneumoniae BTCB04 showed the maximum zone of 25 mm whereas the minimum zone was observed with Pseudomonas aeruginosa BTCB01 at 11 mm. Lastly MTT assay showed no toxicity against cancer cells (HepG2) with AgNPs (0.25-3.5 μM) attaining highest cell viability of 108.5 % with 3.5 μM, whilst Cisplatin was used as a control to complete the test. Cisplatin alone showed viability of 76.3% at 150 μM and 117 % at 100 μM. Synergistic effect between AgNPs and Cisplatin was also investigated, henceforth lowest cell viability of 44.89% was observed with 1:19 combination. In conclusion both of the methods applied for producing silver nanoparticles proved to be successful in reducing the size of AgNPs by various optimization parameters. Although a very small size was obtained by applying conventional one factor at a time method however RSM methodology reduced the size of AgNPs with only 3 parameters which not only lowered the overall cost but labour and time for the experiments. Antimicrobial activity and MTT assay showed significant results and the synthesized AgNPs that can be used in biomedical field in future as they are safe and economic and dependable.
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جیہڑا پنڈ دکھاں دی چا گیا

جیہڑا پنڈ دکھاں دی چا گیا
اوہ بندا مرد سدا گیا
دنیا توں میں مردا ناں
ہجر وچھوڑا کھا گیا
اوہو پکا عاشق اے
جیہڑا توڑ نبھا گیا
جیہڑا پڑھے درود نبیؐ
اوہو رتبے پا گیا
اوہ بھلا نہ بھلا اے
جو شامیں گھر آگیا
جو دل خالص مومن اے
اوتھے رب سما گیا

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Sheikh Walī Ahmad nick named as Sandâkī Baba was a famous political leader, reformer, sūfī saint and literary personality of Swat (KPK), Pakistan. In the subcontinent, he also contributed in freedom movement against the English imperialism like other literary and political workers. Especially he was the founder of Swat State. In this article political, scholarly and reform services of Sheikh Sandâkī has been highlighted.

Distribution Pattern and Conservation Status of Plants Endemic to Pakistan in Hazara Region

This dissertation communicates the results of scientific endeavour regarding the distribution pattern and conservation status of endemic plants of Pakistan distributed in Hazara region (about 20,000 Km2 area) of Northern Pakistan. The study provides a scientific database for developing conservation strategies for threatened endemic taxa of the area. The endemic taxa were determined through a survey of the available literature and herbarium specimens. Organized field surveys were performed during the years 2011-2014 for recording the ground realities and monitoring stations for the taxa conecerned. Actual and potential areas of the distribution of a taxa were thoroughly investigated and locations were georeferenced along the complete field data. Sample of each taxon was properly tagged, identified and preserved. The marked localities were repeatedly surveyed for monitoring fluctuations and decline in a population. Distribution patterns, ecological niche and potential habitats were analysed using Species Distribution Model, Maxent. Twenty Four topoclimatic variables including geology and land cover were used as predictor variables. Distribution of floral associates of endemic taxa were analysed using Twinspan and Nonmetric Detrended Scaling (NMDS). Statistical analyses of predicted habitat maps and floral associates were performed using statistical packages in R. Conservation status of the endemic taxa was determined following IUCN Red List Categories and Criteria (2001, 2012) and guidelines for regional application. Habitat loss was estimated by analysing sixteen days MODIS time series images of thirteen years i.e. 2001-2013, using GIS program IDRISI Selva. Hot spots were identified by calculating the sum of all prediction maps using raster calculator in Arc GIS. Analyses of the generated data revealed that viii endemics were distributed among 28 families 51 genera and 71 species. Boraginaceae and Ranunculaceae were richest families each with 8 endemic taxa followed by Rosaceae (5), Apiaceae (4) and Gentianaceae (4). Genus Pseudomertensia was the largest genus having 8 endemic taxa. Dominant life form among endemics was hemicryptophyte (60) followed phenarphyte (8) and therophyte (2). Highest number were herbs (63) followed by shrubs (7) and trees (1). Maxent AUC range was lying within excellent range (0.92-0.98). Among topographic factors geology was proved to be highly influencial factor affecting the distribution of 39 taxa followed by elevation (28) and land cover (27). Among climatic factors, precipiation of the coldest months was highest dominating factor affecting 31 taxa followed by Mean Diurnal Range (26), mean temperature of the coldest months, temperature annual range (9) and precipitation seasonality (9). While analysing the distribtuon pattern along elevational gradient, highest endemic taxa were found between mid ranges (2000m and 3500m) and lower number was noted at extreme upper and lower ranges. Among the land cover types, highest number of endemics were found in dense temperate mixed forests and dense temperate coniferous forests while least number was found in temperate sparse coniferous forests and subtropical scrub forests and wetlands. With respect to administrative divisions, Mansehra District contained highest endemics followed by Batagram and Kohistan. Among geological classes highest taxa were distrbuted in Proterozoic metaclastic and metasedimentry rocks followed by Proterozoic Cambrian Quartizite and Mezozoic Metasedimentry rocks. Endemic associates were broadly classified into five communities i.e Oxalis-Adiantum-Cymbopogon community, Justicia-Acacia-Cymbopogon community, Trifolium-Pinus-Viburnum community, Valeriana-Salix-Abies community and Poa-Kobresia-Pseudonaphalium community. NDVI revealed that habitat loss was occurring at alaroming rate with 18.3% agricultural extenstion and reduction of forest at 14.7%. Conservation status of endemics revealed that large number of endemics were confined to small geographic ranges with few populations facing multiple threats. The IUCN criteria placed Androsace hazarica, Arabidopsis taraxacifolia, Bupleurum nigrescence, Microsisymbrium falccidum and Neottia inayattii were assigned in Extinct (EX) category, Artemisia amydalina as regionally extinct (RE), Thalictrum secundum ssp. hazaricum and Jasminum leptophyllum as Critically Endangered (CR) at global level and Meconopsis aculeata as Critically Endangered at regional level. Forty taxa got the status of Endangered and 15 were Vulnerable at regional level. Five of the locations were identified as endemic rich areas. It was concluded that endemics were habitat specific and were exposed to a number of threats. Protection of selected sites, restoration of species specific area, recovery of the threatened gene pools and rehabilitation of associated areas needs to be worked out for effective conservation of the endemic plants of the region.