International Scientific School (ISS-2017) Biogenic Silver Composite Polysulfone Membranes with Antimicrobial Activity International Scientific School (ISS-2017) Mach 13-17, 2017 Fozia T. Minhas*, Gulsin Arslan, I. Hilal Gubbuk, Cengiz Akkoz, Betul Yılmaz Ozturk, Baran Asıkkutlu, Ugur Arslan, Mustafa Ersoz

INTRODUCTION Silver nanoparticles (Ag-NPs) are noteworthy because of having prominent optical, biomedical, catalytic, inhibitory and bactericidal properties Mostly Ag-NPs are enormously active for killing food pathogens and meningitis causing microbes. It also bears antifungal property Recently biological methods for Ag-NPs synthesis are adopted mostly owing to non-toxic solvent system, environmental friendly capping and reducing agents, high yield, economical and facile preparation schemes

Ulva compressa (L. ) Kütz. and Cladophora glomerata (L. ) Kütz Ulva compressa (L.) Kütz. and Cladophora glomerata (L.) Kütz. are selected here for the synthesis of Ag-NPs. These are filamentous green macro algae that are abundantly available in natural water streams. Mainly the habitat of these algae are either stony shores or mixed culture growth with other genera such as Cladophora sp. and Ulva sp Furthermore, these synthesized Ag-NPs were blended with PS matrix to evaluate their efficiency as inorganic filter Antimicrobial activity of Ag-NPs/PS composite membrane is examined by Direct contact test (DCT) where close contact between the test microorganism and the tested material was maintained and their influence on microbial viability was analyzed irrespective of the diffusibility and solubility of the antimicrobial species

Overview of the Research Work PHASE-I Ag-NPs Biogenic Synthesis and Characterization of Ag-NPs PHASE-II Ag-NPs/PS Composite Membrane Preparation and Characterization of Ag-NPs/PS Composite Membranes PHASE-III Antimicrobial Activity Direct Contact Test (DCT) Ag-NPs = Silver Nanoparticles, PS = Polysulfone

PHASE-I

Ag-NPs: Synthesis Preparation of Algal Extract Biosynthesis of Ag-NPs PHASE-I Ag-NPs: Synthesis Preparation of Algal Extract Both Ulva compressa (L.) Kütz. and Cladophora glomerata (L.) Kütz. Green seaweed were collected from the Acıgöl Lake, Konya Turkey. The cleaned, dried algal masses were ground into fine powder for further processing. 15 g of each algal powder were taken in 100 ml distilled water at 70 oC with continuous stirring for 24 hr and filtered afterwards. Biosynthesis of Ag-NPs 10 ml of algal extract was poured in clear, transparent solution of 90 ml 0.001 M AgNO3. The colour of AgNO3 solution turned brownish within 6 hr, and formation of Ag-NPs was assured by UV-VIS spectroscopy. The prominent peak at 430 nm confirmed the synthesis of Ag-NPs. Ag-NPs Algae Extract AgNO3

Characterization of Ag-NPs PHASE-I (a) Cladophora based Ag-NPs (b) Ulva based Ag-NPs Characterization of Ag-NPs Au Ag (a) Cladophora based Ag-NPs (b) Ulva based Ag-NPs (b) Ulva based Ag-NPs (a) Cladophora based Ag-NPs (a) Ulva based Ag-NPs (b) Cladophora based Ag-NPs UV-VIS FTIR SEM + EDX AFM XRD

PHASE-II

Preparation of Ag-NPs/PS PHASE-II Preparation of Ag-NPs/PS Composite Membrane The homogenous solution of Polysulfone (PS) was prepared in (15%, w/w) N-methyl-2-pyrrolidinone with constant stirring for 24 hr. Subsequently, 5 mg of Ag-NPs was dispersed in PS solution and coated on glass surface by spin coating technique. Afterwards, coated glass was immersed in deionized water bath and resulting Ag-NPs/PS composite membrane was detached .

Contact Angle Measurement PHASE-II (a) blank PS (b) Cladophora based Ag-NPs/PS Composite Membrane (c) Ulva based Ag-NPs/PS FTIR SEM AFM Contact Angle Measurement (a) blank PS (b) Cladophora based Ag-NPs/PS Composite Membrane (c) Ulva based Ag-NPs/PS 62º 61º 48º (b) Cladophora based Ag-NPs/PS Composite Membrane (c) Ulva based Ag-NPs/PS (a) blank PS Characterization of Ag-NPs/PS Composite Membranes

PHASE-III

Antimicrobial ACTIVITY PHASE-III Antimicrobial ACTIVITY The antimicrobial activity of Ag-NPs/PS composite membrane was evaluated by direct contact method (DCT). In DCT, 96-well microliter plates were utilized and kinetics of bacterial growth in them were analyzed spectrophotometrically at 630 nm. The wall of each “well” was coated with tested material with care so that its floor remain intact. Afterwards, 10 μL bacterial suspension was added above it by keeping “well” in the same position. After incubation period of 1 hr at 37 oC, the suspension liquid evaporated leaving bacteria and surface of targeted material in direct contact. Finally set these plates in spectrophotometer by adding 220 μL of Brain Heart Infusion (BHI) broth. The changes in the optical density of bacterial growth were analyzed periodically after every 60 min at 630 nm for 18 h.

Direct Contact Test (DCT) PHASE-III Direct Contact Test (DCT) P. aeruginosa K. pneumonia E. Coli S.aureus E. faecium Staphylococcus aureus ATCC 25923 Enterococcus faecium ATCC 29212 Escherichia coli ATCC 25922 Pseudomonas aeruginasa ATCC 27853 Klebsiella pneumonia ATCC 700603

CONCLUSION

CONCLUSIONS A green route was adopted to prepare Ag-NPs. Ulva compressa (L.) Kütz. and Cladophora glomerata (L.) Kütz algal extract was used as green source. The Ag-NPs/PS composite was prepared by spin coating technique. Finally, Ag-NPs/PS composite material showed appreciable efficiency against all bacteria i.e. Klebsiella pneumonia, Pseudomonas aeruginasa, Escherichiacoli, Enterococcusfaecium and Staphylococcusaureus in DCT method.

Organizing Committee of ISS-2017 ACKNOWLEDGEMENT Organizing Committee of ISS-2017

in the aqueous extracts of algae Table 1: Preliminary qualitative analysis to examine the phytochemicals in the aqueous extracts of algae ‘+’ and ‘-’ signs indicates the presence and absence of respective components, respectively Phytochemical Constituents U. compressa C. glomerata Molish test + Keller-Killiani Test (Test for Cardiac Glycosides) Fehling’s Test (Test For Reducing Sugars) Alkaloids Terpenoids Flavonoids Tannins Saponins - Amino acids Anthraquinones Phlobatannins

Table 2. Roughness values for Ag-NPs and composite membranes. Sa (nm) Sq (nm) Sz (nm) Ag-NPs (U. compressa) 1.01 1.48 9.09 Ag-NPs (C. glomerata) 0.471 0.848 5.90 PS 20.6 26.5 157 U. compressa based Ag-NPs/PS Composite membrane 11.6 16.1 90.2 C. glomerata based Ag-NPs/PS Composite membrane 9.10 12.4 61.6