2.1 Collection of pathogenic strains
Using a dry cotton swab tip the dermis of the lesions are rubbed and placed in an inverted position in a sterile tube. Sterile cylindrical capped tubes are used to collect the first voiding sputum. The midstream urine is collected in a sterile nonporous container. An unbreakable screw cap bottle integrated with a spoon is used to collect the mucous or blood coated stools. Sterile IV needles are used to drain blood samples which are then loaded in a tube covered with EDTA to avoid clotting [5]. Name, age, sex, color, and texture of the sample using waterproof pen legibly noted on the sealed sample and placed them in an icebox and shifted to the laboratory within 6 hrs. All these samples are collected from different wards of various hospitals and laboratories in Madurai. Pre-collection techniques are followed properly to isolate the suspicious pathogen and cross-contamination is prevented by wearing gloves, a mask lab coat, and a head cap [6].
2.2 Isolation and screening
MacConkey agar, Nutrient agar, blood agar, and mannitol salt agar, are prepared in appropriate concentration than using an incinerated loop or sterile dry cotton swab gently touch the surface of the exudates in the collection tube than by zig-zag, perpendicular, and quadrant method of streaking inoculation done in their corresponding labeled plate [7]. Texture, color, size of each colony is noted after 12-24 hrs of incubation then after incineration of the loop a colony is rubbed on the slide and showed in the flame for smear preparation. Then the smear is wetted by crystal violet, mordant, acetone, and at last safranin pro a minute and cleansed in water, air dried, and observed under the microscope to observe whether it is a gram-positive or gram-negative bacteria. IMVIC, oxidase, urease, mannitol, and citrate are the confirmatory biochemical test done to identify the above isolates which show a color change by adding reagents like methyl red Kovacs reagent, glucose broth, etc [8].
2.3 Floral collections
A cost-effective, viridescent, stalkless, perennial herb is plucked in the ponds, and canals from the nearby villages of Rasipuram [9]. Then surface sterilization is done by rinsing in water and separated the affected leaf. Then clean, unaffected leaves are shadow dried, pulverized and made into fine crystals for extraction.
2.4 AS-AgNPs synthesis
After finding the dry weight of the fine crystals they are treated with aqua and then stirred continuously into a concentrated solution. After adding in a small beaker they are placed in a heating mantle and parameters optimization is done [10]. Then the precursor silver nitrate at a concentration of 1 Mm is mixed gently. In the absence of light after 72 hrs a color change from straw to chocolate brown was monitored at regular intervals this indicates the presence of silver atoms in the process and finally, the collected residues from the Whatman filter paper No1 are kept in the hot air oven at a temperature of 80°C to remove the water droplets completely. Later those granules are stored for further standard confirmatory analysis [11].
2.5 Antibacterial activity
In prepared MHA plate using a sterile dry cotton swab tip touched the surface of the isolated colony gently and streaked completely all over the plate then after incineration of the cork Borell, 6mm a punch was made on the plate [12, 13]. Then all the plugs are removed aseptically using forceps or loop. Different concentrations of AS-AgNPs crude (such as 20µg, 40µg, and 60µg) using micropipette loaded on each well. After inoculation, the plates are placed in the upright position in the incubator for 24 -48 hrs [14].
2.6 Fabric sensitivity test
In the freshly prepared muller, Hinton agar plate the pathogens are spread evenly by using a sterile dry swab, and then fabrics such as cotton and mixed cotton are made into small square pieces soiled in crude of AS-AgNps placed in the plate [15]. Then a non coated AS-AgNps fabrics act as a control.
2.7 Confirmational study of AS-AgNPs coated on fabrics
i) UV- Visible spectroscopy
The silver nanoparticles synthesis from the phyto extract processed in a DBPEL (double-beam Perkin-Elmer Lambda) UV/Vis spectrometer and all the processes take place in fused silica glass cuvette under magneto stirring.
ii) FTIR
FTIR is used to establish the functional groups, absorption, and efficiency between different components of the synthesized product is noted [16]. After preparing the sample it is placed in the infrared beam using a sample holder then after many scans are taken and its resolution, the spectral range is measured.
iii) SEM with EDAX
It is used to generate the corresponding electron from the electron detector and also generate the surface topological image of the specimen at elevated magnification [17]. Samples are prepared by cutting into thin films using diamond abrasives and mounted if the surface is rough they are scratched out using sandpaper or pumice stone and cleansed using solvents like methanol, ethanol, and acetone, and finally, thermal etching analysis is done.
The energy distribution and photon of light focused on the surface of the specimen by Energy-dispersive X-ray spectrometers and is used to measure the metal nanoparticles qualitatively and quantitatively in the material.
iv) HR-TEM
A crystallographic structure of the image at an atomic level is recorded at high magnification and resolution with the help of a beam of electrons [18]. The sample is made into thin sections using diamond abrasives, mounted, and finally, the image in nanometer-scale against the black and white background is observed.