This document discusses nanoparticles and summarizes a study on the biosynthesis of silver nanoparticles (AgNPs) using the fungus Rhizopus stolonifera. The study involved isolating R. stolonifera from tomato fruits and producing biomass through cultivation. The biomass was then used to synthesize AgNPs when mixed with silver nitrate solution. Formation of AgNPs was indicated by a color change from white to yellow. UV-vis spectroscopy showed a peak at 435nm, confirming nanoparticle synthesis. The synthesized AgNPs were tested against several bacterial strains through a well diffusion assay. The AgNP pellet demonstrated zone of inhibition, indicating antibacterial activity against Salmonella typhi, Escherich

1. NANOPARTICLES
ASIF NAWAZ
MPHIL 2ND
ABBOTTABAD UNIVERSITY OF SCIENCE AND TECHNOLOGY

2. OUTLINES
• introduction
• Methods used for synthesis of NPs
• Biological synthesis NPs
• AgNPs biosynthesis using R.Stolonifera
• Applications

3. INTRODUCTION
• “There is plenty of room at the bottom”
(Richard Feyman 1959)
• Norio Taniguchi coined the word “nanotechnology” for the first time in 1974
• Nano' derives from the Greek word “nanos” which means dwarf or extremely small
• It can be used as a prefix for any unit to mean a billionth of that unit 10−9
• technologies, that measure, manipulate, or incorporate material or features with at least one critical
dimension between ~ 1 nanometer and 100 nanometers is called nanotechnology
• whose applications exploit properties, distinct from bulk/macroscopic systems, that arise from their
scale/critical dimension

4. DIFFERENT NANOPARTICLES
• Some nanoparticle are listed bellow with potential of antibacterial activity
• Gold NPs
• Iron oxide NPs
• Silver NPs
• ZnO NPs

5. METHODS USED FOR SYNTHESIS OF NANOPARTICLE
Physical
• Ball Milling
• Thermal evaporation
• Vapour Phase
Chemical
• Sol-gel Processing
• Solution-based synthesis
Biological
• In bacteria
• In fungi
• Yeast
• Using plant extracts

6. BIOSYNTHESIS OF NANOPARTICLE
Plant & Plant extract
• Extracellular Inexpensive reduction
Fungi and Yeast
• Extra cellular
• Intera cellular
• Cell wall surface
Bacteria
• Extracellular
• Intracellular

7. NANOPARTICLE SYNTHESIS BY PLANT EXTRACT

8. SILVER NANOPARTICLE SYNTHESIS USING FUNGUS
RHIZOPUS STOLONIFERA
Isolation of Rhizopus stolonifera
• R. stolonifera was isolated from naturally infected tomato fruits by rubbing swab and inoculated onto
Potato dextrose Agar (PDA) medium supplemented with (30 mg/L) chloramphenicol to discourage
bacterial contamination was prepared routinely and used for fungal isolation
• The sample was serially diluted and were grown on PDA and were identified using cultural and
morphological characteristics using compound microscope
Biomass production for AgNPs synthesis
• Biomass was produced by cultivation of R. stolonifera in Malt Glucose peptone (MGYP) broth and
Culture was incubated at 40 °C on an orbital shaker 180 rpm for 3 days

9. • Culture was filtered and the resulted biomass was washed extensively by deionized water to get rid of
adhered media parts
• Mycelia extract was prepared by suspension of fungal biomass in 100 ml deionized water and incubated as
described above for 72 h
AgNPs synthesis
• After that mycelia suspension was filtered using (Whatman paper No. 1). The Resulted filtrate (mycelia
extract) was mixed with AgNO3 solution (1 mM AgNO3 final concentration) and incubated on orbital shaker
180 rpm at 40°C for two days
• A control containing a 1mM AgNO3 solution and extract was incubated separately in same conditions in
parallel
• After incubation the change in control from whitish transparent to pale yellow was observed in test sample as
compared to control which indicate the formation of silver NPs

10. NANOPARTICLES SYNTHESIS INDICATED BY CHANGE IN COLOR AFTER
48 HOUR B AND A CONTAIN ONLY MYCELIAL FILTRATE

11. • Further the sample was examined by labomed UV-visibles spectrophotometer, 200-700 wavelength
beam was passed from the sample
• The absorption graph was at peak between 400-500nm rang while the best peak was observed at 435
Which is the evidence of biosynthesis of silver nanoparticles
Antimicrobial Activity of synthesized AgNPs
• To check the antibacterial of silver nanoparticle the sample was centrifuged at 10,000 Rpm for 10
minutes to remove the fungal extract in supernatant and the NPs were settled down to the pellet
• The pellet was then mixed with deionized water and varied concentration was inoculated into freshly
prepared plates of Salmonella typhi, Escherichia coli, Citrobacter, staphylococcus aureus and pseudom
aeruginosa by well diffusion method and were incubated at 37°C for 16-18 hours and then the zone of
inhibition was measured,

12. UV-VIS ABSORBANCE MAXIMA OF SILVER NANOPARTICLES
SYNTHESIZED BY RHIZOPUS STOLONIFERA

13. SENSITIVITY REPORT OF SELECTED BACTERIAL STRAINS TO 30ЦL,
50ЦL PELLET AND 50ЦL SUPERNATANT OF AGNPS
Test
organism
Salmonella
typhi
Citrobacter Escherichia
coli
Pseudomonas
aeruginosa
Staphylococcus
aureus
Pellet(30цl) 12±1mM 1o±1mM 5±1mM 11±1mM 12±1mM
Pellet(50цl) 16±1mM 12±1mM 8±1mM 13±1mM 14±1mM
Super
natant(50цl)
8±1mM 2±1mM 3±1mM 2±1mM 2±1mM

14. APPLICATIONS OF SILVER NANOPARTICLE