TECHNICAL FIELD

[001] The present disclosure relates to development of antimicrobial product especially against MRSA, VRSA and Pseudomonas aeruginosa activity. The present disclosure relates to a textile material treated with one or more bioactive isolates extracted from microfungi, macrofungi (mushrooms), actinomycetes, bacteria and plants to provide antimicrobial activity especially against MRSA, VRSA and Pseudomonas aeruginosa, and the textile fabrics will be utilized as medical textiles (Meditex) including for preparing various hospital products and garments.

BACKGROUND

[002] MRSA is a type of bacteria that is resistant to a number of widely used antibiotics. In other words, MRSA infections can be more difficult to treat than other bacterial infections. The full name of MRSA is methicillin-resistant Staphylococcus aureus. It is also called as a "superbug." Staphylococcus aureus (also known as staph) is a common type of bacteria.

[003] Vancomycin-resistant Staphylococcus aureus (VRSA) is the term used to describe the strains Staphylococcus aureus (SA) that are fully resistant to the antibiotic vancomycin. The expected emergence of VRSA is alarming because vancomycin is the only antibiotic that is effective against MRSA, strains of SA that are resistant to the antibiotic methicillin (MRSA).

[004] Pseudomonas aeruginosa has become an important cause of gram- negative infection, especially in patients with compromised host defense mechanisms. It is the most common pathogen isolated from patients who have been hospitalized longer than 1 week, and it is a frequent cause of nosocomial infections. P. aeruginosa can develop resistance to antibacterial agent either through the acquisition of resistance genes on mobile genetic elements (i.e., plasmids) or through mutational processes that alter the expression and/or function of chromosomally encoded mechanisms. P. aeruginosa uses multiple methods to resist the effect of antibiotics, including pumping the drugs out of its cell matrix, production of antibiotic inactivating enzyme and it is constantly evolving new methods of resistance. The bacteria are highly common, living in water, the earth, man-made surfaces and even in our skin. When tissue is damaged or the immune system is not working well, the bacteria strikes, infecting the body and causing sepsis and inflammation. It can survive on medical equipment such as catheters, and the bacteria are often found on to have aggressively remained on other hospital supplies - even after cleaning - leading to its transmission from patient to patient. If the bacteria colonize vital body organs such as the lungs, death can occur. The purpose of imparting antimicrobial activity to textiles is to protect the textile material from microbial attack, prevent the transmission, and spreading of pathogenic microorganisms, inhibit odor development resulting from microbial degradation, and creating a textile material that will act as preventive and/or curative treatment against multidrug resistant or pathogenic microorganisms.

[005] The abuses of antibiotics, new virus occurrence, and prevalence of multi- resistant bacteria, such as Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE), along with ageing of the population have generated high consumptions of medical textiles. Many commercially-used antimicrobial products are textiles treated with e.g. silver, triclosan, polyhexamethylene biguanide (PHMB) and quaternary ammonium compounds to provide resistant to various microbes. However, the above materials are biocides.

[006] Biocide means (as per European legislation) a chemical substance or microorganism intended to destroy, deter, render harmless, or exert a controlling effect on any harmful organism by chemical or biological means. The US Environmental Protection Agency (EPA) uses a slightly different definition for biocides as "a diverse group of poisonous substances including preservatives, insecticides, disinfectants, and pesticides used for the control of organisms that are harmful to human or animal health or that cause damage to natural or manufactured products " In general, the term roughly includes plant protection products and some veterinary medicines. [007] The microbial infection is carried on the skin and inside the nostrils and throat, and can cause mild infections of the skin, such as boils and impetigo. If the bacteria get into a break in the skin, they can cause life-threatening infections, such as blood poisoning or endocarditis. MRSA bacteria are usually spread through skin-to-skin contact with someone who has an MRSA infection or has the bacteria living on their skin. The bacteria can also spread through contact with contaminated objects such towels, sheets, clothes, dressings, surfaces, door handles, and floors. [008] People staying in hospital are most at risk of becoming infected with MRSA because:

They are surrounded by a large number of people, which means the bacteria can spread more easily;

- They often have an entry point for the bacteria to get into their body, such as a surgical wound or urinary catheter; and

They may have serious or complex health problems, which makes them more vulnerable to infection.

[009] It is also possible to people become infected with MRSA outside of hospital, although this is much less common.

[0010] In recent years, rates of MRSA have fallen because of increased awareness of the infection by both medical staff and the public. However, MRSA still places a considerable strain on healthcare services.

[0011] An antimicrobial treated material is defined as being hygienic and, therefore, should have the following requirements (Gao Y, Cranston R. Recent Advances in Antimicrobial Treatments of Textiles. Textile Research Journal. 2008; 87:60-72. & Hashem M, Ibrahim NA, El-Sayed WA, El-Husseiny S, El- Enany E. Enhancing antimicrobial properties of dyed and finished cotton fabrics. Carbohydrate Polymers. 2009; 78:502-510. - Effective inhibition against a broad spectrum of bacterial and fungal species,

- Non-toxicity to the consumer, manufacturer and the environment,

- Durability,

- Compatibility with resident skin microbiota, and other finishing processes, - Avert from irritations and allergies,

- Applicability with no adverse effects on the quality or appearance of the textile.

Antimicrobial agents can act in two distinct ways

[0012] (i) by contact; the antimicrobial agent inhibits microbes only on the fibre surface (substances are permanently attached to the fibre surface), & (ii) by diffusion; the antimicrobial agent is slowly released onto the fibre surface and/or from the surface (i.e. substances with controlled-release mechanism).

[0013] There is a vast resource of natural antimicrobial agents, which can be used for imparting antimicrobial property to textile substances. Although, there are many cited literature, wherein efforts have been made to exploit these eco- friendly bioactive natural products for textile application, but there are very few studies which carries out systematic in-depth investigation. The major challenges in the application of natural products for textile application are the most of these bio materials that are complex mixtures of several compounds and also the compositions vary in different species. The activity of the composition varies, depending on their geographical location, age, and method of extraction. The availability of these products in bulk quantities, their extraction, isolation, and purification to get standardized products are other challenges in their application. The durability, shelf life and antimicrobial efficiency vis-a-vis synthetic agents are others issues, which need to be looked into. However, none of the literature reported for MRSA, VRSA and Pseudomonas aeruginosa activity from eco- friendly and non-toxic properties of bioactive isolates which not only provides MRSA, VRSA and Pseudomonas aeruginosa activity also act as a coloring agents for textiles. These textiles could be woven, nonwoven, knitted, composites etc. and have found different end-uses in medical and healthcare applications.

[0014] Depending on the specific end-use, different products have to meet the demands for the specific end-use performances. Irrespective of their applications, internal (surgical threads and various implants) or external (gauzes, bandages, surgical masks, gowns and apparel, nappies, tampons, and so on), medical textiles have to be comprised of basic bioactive properties to provide MRSA and VRSA activity.

[0015] Similarly, it is extremely important that protective clothing such as surgical masks, caps, gowns, etc. and hospital linen meet the demands for MRSA and VRSA or other multi resistant active bacteria like P. aeroginosa.

[0016] The materials for use in surgery have to ensure adequate protection against microorganisms, biological fluids, and aerosols, i.e. impermeability for microorganisms in wet and dry atmospheres, and for air-borne microorganisms. Further, disease transmission prevention is a very important consideration for intracorporeal or implantable devices within the human body (e.g. vascular grafts and sutures) and for extracorporeal devices such as catheters and hollow fibres for dialyzers. Furthermore, wound dressings also need to prevent infection and promote faster wound healing. Therefore, controlling the undesirable effects of microorganisms on textiles is becoming an important issue, especially within the medical textile industry. SUMMARY OF THE INVENTION

[0017] Accordingly, the present disclosure relates to an antimicrobial material comprising a textile material treated with one or more bioactive isolates. The bioactive isolate is extracted from a biological material selected from a group consisting of macro fungi, micro fungi, actinomycetes, bacteria and plants.

[0018] Another aspect of the present disclosure relates to a method of preparing the bioactive isolate by extracting the biological material with one or more solvent and filtering the mixture to remove solid content to get a solution. This solution may be directly used for treating the textile material or the solution is dried to obtain a powder.

[0019] One more aspect of the present disclosure relates to a method for treating textile material to impart antimicrobial or biocidal properties thereto, the method involves treating the textile material with a solution of bioactive isolates as described above. DESCRIPTION OF THE INVENTION

Definition of terms:

[0020] Here, technical terms and scientific terms used in the present specification have the general meaning understood by those skilled in the art to which the present invention pertains unless otherwise defined.

[0021] The articles "a", "an" and "the" are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article.

[0022] The terms "comprise" and "comprising" are used in the inclusive, open sense, meaning that additional elements or steps may be included. [0023] The term "including" is used to mean "including but not limited to". "Including" and "including but not limited to" are used interchangeably.

[0024] In the present disclosure, the term antimicrobial activity is referred to MRS A resistant activity, VRSA resistant activity and other multidrug resistant bacteria including P. aeruginosa, Vancomycin Resistant Enterococci, Klebsiella pneumoniae, Streptococcus pneumoniae, Salmonella sp., Escherichia coli, extensively drug resistant tuberculosis, Acinetobacter baumannii, Neisseria gonorrhaeae, Clostridium difficile, and Candida sp. related infections.

[0025] Bioactive isolate means, a substance obtained from a biological material by extracting the biological material with one or more solvent. The above extract may be in any form or any color. The bioactive isolate may also refer to as bioactive metabolite in the description.

[0026] Nosocomial means originating or taking place in a hospital, acquired in a hospital, especially in reference to an infection. [0027] Actinomycetes_are a specific group as bacteria. Morphologically they resemble fungi because of their elongated cells that branch into filaments or hyphae.

[0028] Padding method: Padding technique is widely regarded as a textile finishing technique, can be used to add a variety of coatings, but this usually refers to a fibre coating for the application of micro or nano materials or chemical compositions. The fabric is submerged in the coating solution then the excess solution squeezed out in the rollers, which dictates the pick-up percentage, the fabric is then dried and cured/finished. [0029] Exhaust method: Exhaust Dyeing is one of the most popular dyeing methods. Most of the dyes could be used for exhaust dyeing of textile material. A dye solution or dye bath is produced by dissolving the dyestuff according to required liquor ratio. Then the textile material is immersed into the dye solution. Initially the surface of the fibre is dyed when dyes contact with the fibre, then the dyes are entered in the core of fibre. Proper temperature and time is maintained for diffusion and penetration of dyes molecule in the fibres core. In exhaust dyeing, all the textile material contacts all the dye liquor and the fibres absorb the dyes. The concentration of the dye in the dye bath therefore gradually decreases. [0030] Jigger Dyeing Machine for Woven Fabrics: Dyes use for dyeing fabric in Jig or Jigger dyeing machine which is suitable for woven fabric as the jigger exert considerable lengthwise tension on the fabric. This is also particularly suitable for fibers because the natural dyes generally do not exhaust well and the jig works with an exceedingly low liquor ratio. It is one of the oldest dyeing machines for dyeing fabric. There are mainly two types of jigger dyeing machine for dyeing woven fabrics with dyes. First type is open jigger dyeing machine and the other is closed jigger dyeing machine.

[0031] Accordingly, the present disclosure is focused on the production, extraction of different bioactive isolates from different biological sources and applied to the textile for example cotton, silk, wool or synthetic fabrics to impart antimicrobial activity to the textile materials. The disclosure also includes textile material coated previously with chemically dyed fabrics further treated with the bioactive isolate of the present disclosure to provide antimicrobial activity against MRSA, VRSA, P. aeruginosa, Vancomycin Resistant Enterococci, Klebsiella pneumoniae, Streptococcus pneumoniae, Salmonella sp., Escherichia coli, extensively drug resistant tuberculosis, Acinetobacter baumannii, Neisseria gonorrhaeae, Clostridium difficile, and Candida sp. related infections [0032] The present disclosure also provides various treatment methods followed in treating the textile material or textile fabric with the bioactive substances. Further, the prepared fabric will also extend towards other multi resistant organisms.

[0033] The textiles material which, were applied with bioactive isolates on fabrics were tested at different time intervals and at different washing periods to evaluate the antimicrobial activity.

[0034] The present disclosure provides an antimicrobial material comprising a textile material; and one or more bioactive isolates, wherein the textile material is treated with said one or more bioactive isolates, wherein said bioactive isolate is extracted from a biological material selected from a group consisting of macrofungi, microfungi, actinomycetes, bacteria and plant or plant parts thereof selected from Oldenlandia umbellata, and Indigofera sp.

[0035] In one embodiment of the present disclosure provides a treated textile material imparted with antimicrobial or biocidal properties against MRSA or against VRSA or both.

[0036] In another embodiment of the disclosure provides a treated textile material which is capable of providing resistance against Pseudomonas. aeruginosa, Vancomycin Resistant Enterococci, Klebsiella pneumoniae, Streptococcus pneumoniae, Salmonella sp., Escherichia coli, extensively drug resistant tuberculosis, Acinetobacter baumannii, Neisseria gonorrhaeae, Clostridium difficile, and Candida sp. related infections.

[0037] In one implementation of the present disclosure provides an antimicrobial textile material selected from cotton, silk, polyester, rayon, nylon, wool, acrylic, pre-printed textile fabric, woven-fabric, dyed-woven fabric, raw fiber, non-woven fabric, silk fiber, silk fabric, synthetic textile, wound dressing, and yarn.

[0038] In one implementation, the biological material is a macrofungi, which is selected from Ganoderma lucidum (GL) and Pycnoporus sanguineus (PY). [0039] In one implementation, the biological material is a microfungi, which is selected from Hypocrea rufa (Hr), Phoma foveata (PH), Curvularia sp. (Cur), Fusarium sp. and Pestalotiopsis sp.

[0040] In one implementation, the biological material is a actinomycetes and is selected from Streptomyces species such as Streptomyces sp. (unidentified) Streptomyces cacaoi, and Streptomyces parvulus.

[0041] In one implementation, the biological material is bacteria, which is selected from Pseudomonas sp., Chromobacterium sp. and Serratia marcescens.

[0042] In one implementation, the biological material is a plant or plant parts thereof, selected from Oldenlandia umbellata, and Indigofera sp. [0043] In another implementation, the bioactive isolate is obtained from extracellular material of bacteria, microfungi and actinomycetes. [0044] In another implementation the bioactive isolate is obtained from intracellular material of bacteria and macro fungi.

[0045] In another implementation provides an antimicrobial textile material treated with bioactive isolate, wherein the amount of the bioactive isolate used for treating one-unit area (1 sq.mtr) of the textile material is in the range of 0.2 g to 2.0 g per one square meter of the textile material.

[0046] In another implementation provides an antimicrobial textile material treated with bioactive isolate, wherein ratio of textile material to the bioactive isolate is in the range of 1: 10 to 1:50. [0047] Yet another implementation of the disclosure provides an antimicrobial textile material which is nonirritant to skin.

[0048] Yet another implementation of the disclosure provides an antimicrobial textile material which is stable in terms of its antimicrobial activity at a temperature of more than 100°C or under sterilization conditions. The treated textile material retains its antimicrobial resistance after subjecting the textile material to a temperature beyond 120°C during sterilization before the use in hospital etc.

[0049] Yet another implementation of the disclosure provides an antimicrobial textile material to produce hospital textile products such as surgical gowns, mask, caps and bandage.

[0050] Yet another embodiment of the present disclosure provides a method of preparing the bioactive isolate from biological material as recited in previous paragraphs. The method of preparing the bioactive isolates involves: obtaining a mixture of one or more biological material by adding one or more solvent to the biological material to obtain a mixture;

stirring the mixture with a solvent for a period ranging between 10 minutes to 120 minutes at a temperature between 10°C to 100°C to obtain a stirred mixture; and

filtering the stirred mixture, to obtain a bioactive isolate solution;

[0051] Yet another embodiment of the present disclosure provides a method of preparing the bioactive isolates involves:

obtaining a mixture of one or more biological material by adding one or more solvent to the biological material to obtain a mixture;

stirring the mixture with a solvent for a period ranging between 10 minutes to 120 minutes at a temperature between 10°C to 100°C to obtain a stirred mixture; and

filtering the stirred mixture, to obtain a bioactive isolate solution; and - drying to obtain powdery bioactive isolate.

[0052] Yet another embodiment of the disclosure provides a method of preparing the bioactive isolate by drying the filtered bioactive isolate solution in order to remove the volatiles present in it. After filtering the bioactive isolate solution, the bioactive isolate may contain volatiles which is removed by drying the bioactive isolate at a temperature between 60°C and 80°C to get the bioactive isolate as a dry powder.

[0053] In yet another embodiment of the disclosure relates a method of preparing bioactive isolate wherein enhanced recovery of the bioactive isolate from the biological material by repeating the solvent extraction step by several times i.e. about 4 to 7 times.

[0054] In an another embodiment of the present disclosure, the solvent used for extracting bioactive isolate from the biological material is selected from water, alcohol, aqueous alcohol, chloroform, and acetone or mixtures thereof. Preferred solvent for the extraction is water. The amount of solvent used for extraction is in the range of 1 to 15 times to the weight of the biological material used. The same amount of solvent is used for repeated extraction of bioactive isolate in order to enhance the recovery of the bioactive isolate. [0055] In another embodiment of the present disclosure provides a method for isolating the bioactive isolate by cultivating the specific biological material using appropriate media and harvesting the biomass / mycelia. The bioactive isolates from macrofungi were grown in different substrates and harvested the basidiomata. The extracellular bioactive isolate from microfungi or bacteria was filtered and the intracellular bioactive isolate of macrofungi or bacteria were extracted with solvent selected from water, hot water, alcohol such as methanol, ethanol and propanol, acetone, chloroform and extracted to obtain bioactive isolates. The bioactive isolate is separated by simple liquid: liquid partition method if required. [0056] Yet another embodiment of the present disclosure relates a method for treating a textile material in order to impart antimicrobial, biocidal properties thereto. The method of imparting resistance against MRSA or VRSA activity or resistance to other drug resistance pathogens, super bug etc. involves treating the textile material with one of more bioactive isolate as described above.

[0057] In an implementation the method for treating a textile material in order to impart antimicrobial, biocidal properties thereto includes (a) preparing a solution of a bioactive isolate by mixing the bioactive isolate in a solvent; (b) treating the textile material with the solution of the bioactive isolate to achieve a desired amount of solution pickup; (c) drying the textile material for removing volatiles and (d) finishing of the textile material as an antimicrobial fabric.

[0058] In another implementation, the method for treating a textile material in order to impart antimicrobial, biocidal properties thereto includes (a) preparing a solution of a bioactive isolate by mixing the bioactive isolate in a solvent; (b) treating the textile material with the solution of the bioactive isolate to achieve a desired amount of solution pickup; (c) drying the textile material for removing volatiles; (d) adding a fixing agent; and (e) finishing of the textile material as an antimicrobial fabric.

[0059] In another implementation, the method for treating a textile material in order to impart antimicrobial, biocidal properties thereto, the bioactive isolate is obtained from a biological material selected from a group consisting of macrofungi, microfungi, actinomycetes, bacteria and plant or plant parts thereof selected from Oldenlandia umbellata, and Indigofera sp.

[0060] In another implementation, the method for treating a textile material in order to impart antimicrobial, biocidal properties thereto, the textile material is selected from cotton, silk, polyester, wool, rayon, nylon, pre-printed textile fabric, woven-fabric, dyed-woven fabric, raw fiber, non-woven fabric, silk, synthetic textile or bandage cloth, woven fabric or non- woven fabric and yarn.

[0061] In yet another implementation, the method for treating a textile material in order to impart antimicrobial, biocidal properties thereto, no mordant is used for treating the textile material and the textile material is directly treated with bioactive isolate solution.

[0062] In yet another implementation, the method for treating a textile material in order to impart antimicrobial, biocidal properties thereto, the textile material is treated with a mordant before the treatment of the textile material with bioactive isolate solution. The mordant used is selected from tannic acid, alum, chrome alum, sodium chloride, salts of aluminium, chromium, copper, iron, iodine, potassium, sodium, tungsten and tin.

[0063] In yet another implementation, the method for treating a textile material in order to impart antimicrobial, biocidal properties thereto, treating the textile material with the solution of the bioactive isolate is carried out by either padding method or exhaust method.

[0064] In yet another implementation, the textile material is treated with the bioactive isolate is carried out using jigger machine for the exhaust method.

[0065] In yet another implementation, the textile material is treated with the bioactive isolate is carried out using padding machine.

[0066] In yet another implementation the method for treating a textile material in order to impart antimicrobial, biocidal properties thereto, powdery bioactive isolate is dissolved in a solvent before the treatment of the textile material. The solvent used may be organic solvent or water which depends on the solubility of the bio active isolate.

[0067] In case if the bioactive isolate is in solution form after filtering the solid portion, the bioactive solution is used as such for treating the textile material for imparting antimicrobial activity. For example, in the present disclosure the extracellular portion of the biological material, after cultivating the fungi using a media, the media as such is used after removing the solid particles.

[0068] In another embodiment of the present disclosure relates to stability characteristics of the treated textile material after repeated washings as per the industrial standards. A textile fabric treated with the GL bioactive isolate is washed for 30 times and even after 30 washes the treated fabric retains the antimicrobial activity against MRSA or VRSA.

[0069] In another embodiment, the treated textile material was evaluated for its antimicrobial activity and it was found that even after storing the treated textile material under normal conditions for a year, the textile material retained its antimicrobial activity.

[0070] In another embodiment, the bioactive isolate was stored for a year and used for treating the textile material for imparting antimicrobial activity. The treated textile material was tested for antimicrobial activity and found no change in the activity.

[0071] List of tables:

1. Table 1: List of microbes used and type of extract used to obtain bioactive isolates. 2. Table 1A - Details of bioactive isolate used in treating the textile material.

3. Table-2 Evaluation of treated textile material using various bioactive isolates against MRS A resistant activity as mentioned in in examples 1- 7

4. Table 3: Stability of the antimicrobial activity especially against MRS A of the textile material treated with the microbial bioactive isolates stored for a year period.

5. Table 4: Evaluation of bioactive isolates treated textile material for MRS A resistant activity- after one hour

6. Table 5: Evaluation of antibacterial textile material especially against MRSA and VRSA - after a year period.

7. Table 6 Evaluation of antibacterial textile material for MRSA, VRSA and Pseudomonas aeruginosa resistant activity - after one and half year period.

[0072] Table 1: List of microbes used and type of extract used to obtain bioactive isolates.

Curvularia htnala(Cm) Extracellular

Curvularia sp. Extracellular

Fusarium sp. Extracellular

Pestalotiopsis sp. Extracellular

Actinomycetes cultures

Streptomyces sp. (unidentified) Extracellular

Streptomyces cacaoi Extracellular

Streptomyces sp. (unidentified) Extracellular

Streptomyces parvulus Extracellular

Extraction of bioactive isolates for preparation of antimicrobial textiles [0073] Extraction 1: Cultivated Pycnoporus sanguineus (PY) basidiomata using different substrates was made in to dry powder form and the same was extracted with hot water (100°C). Successive extraction was continued till the bio mass became fade or the extract become colorless. After extraction, the solution was filtered to remove solid portion of the biological material and the extracted bioactive mixture was converted into powder form using hot air oven (60 -80 °C), Spray dryer and also freeze drying. Finally, 470 g of the bioactive isolate was used for preparation of 100 m of antibacterial textile material. [0074] Extraction 2: Cultivated Ganoderma lucidum (GL) basidiomata using different substrates was made in to dry powder form and the same was extracted with hot water (100 °C). Successive extraction was continued till the biomass became fade or the extract become colorless. After extraction, the solution was filtered to remove solid portion of the biological material and the extracted bioactive mixture was converted into powder form using hot air oven (60 -80 °C), Spray dryer and also freeze drying. Finally, 450 g of the bioactive isolate was used for preparation of 100 m of antibacterial textile material. [0075] Extraction 3: Production of extracellular bioactive isolates from Phoma foveata (PH) was carried out in the optimized nutrient media, and the mixture was filtered to remove solid portion of the biological material and the extracted bioactive mixture was converted into powder form using hot air oven (60 -80 °C) or Spray dryer or freeze drying. Finally, 210 g of the bioactive isolate was used for preparation of 100 m of antibacterial textile material.

[0076] Extraction 4: Production of extracellular bioactive isolates from Hypocrea rufa (Hr) was carried out in the optimized nutrient media, and the mixture was filtered to remove solid portion of the biological material and the extracted bioactive mixture was converted into powder form using hot air oven (60 -80 °C) or Spray dryer or freeze drying. Finally, 250 g of the bioactive isolate was used for preparation of 100 m of antibacterial textile material.

[0077] Extraction 5: Production of extracellular bioactive isolate from Curvularia lunata (Cur) was carried out in the optimized nutrient media, and the mixture was filtered to remove solid portion of the biological material and the extracted bioactive mixture was converted into powder form using hot air oven (60 -80 °C) or Spray dryer or freeze drying. Finally, 90 g of the bioactive isolate was used for preparation of 100 m of antibacterial textile material.

[0078] Extraction 6: Production of extracellular bioactive isolates from Streptomyces parvulus (SP) was carried out in the optimized nutrient media, and the mixture was filtered to remove solid portion of the biological material and the extracted bioactive mixture was converted into powder form using hot air oven (60 -80 °C) or Spray dryer or freeze drying. Finally, 300 g of the bioactive isolates was used for preparation of 100 m of antibacterial textile material. [0079] Table 1A - Details of bioactive isolate used in treating the textile material.

Width of the fabric is 2.90 meters

[0080] Example 1:

Cotton fabric treated with tannic acid fixed with bioactive isolates from PH recorded the MRSA activity of 96% at 24-hours.

Treatment: Cotton fabric pretreated with tannic acid and then fixed with PH bioactive isolates (PHT) by exhaust method.

Organism: MRSA (ATCC 33591); Method: ASTM E2149-10

Activity: 96% (24 hours);

Remarks: Excellent antimicrobial activity

[0081] Example 2:

Cotton fabric applied with alum and the bioactive isolates of GL using the padding method of application recorded the antibacterial activity of 88 % at 24 hours against MRSA. Treatment: Cotton fabric pretreated with alum and then padded with GL bioactive isolates (GLA).

Organism: MRSA (ATCC 33591); Method: ASTM E2149-10

Activity: 88% (24 hours);

Remarks: Excellent antimicrobial activity

[0082] Example 3:

Cotton fabric treated with tannic acid when applied with the PH bioactive isolate using padding method recorded antibacterial activity of 81 % at 24 hours against MRSA

Treatment: Cotton fabric pretreated with tannic acid and then padded with PH bioactive isolates (PHT).

Organism: MRSA (ATCC 33591); Method: ASTM E2149-10

Activity: 81% (24 hours);

Remarks: Excellent antimicrobial activity

[0083] Example 4:

Cotton fabric treated with alum when applied with the GL bioactive isolate using exhaust method recorded the antibacterial activity of 88 % at 24 hours against MRSA.

Treatment: Cotton fabric pretreated with alum and then fixed with GL bioactive isolates (GLA) by exhaust method

Organism: MRSA (ATCC 33591); Method: ASTM E2149-10

Activity: 88% (24 hours);

Remarks: Excellent antimicrobial activity

[0084] Example 5: Cotton fabric treated with tannic acid and bioactive isolates of PH and then combined with GL recorded antibacterial activity of 89 % at 24 hours against MRSA

Treatment: Cotton fabric pretreated with tannic acid and fixed with PH bioactive isolate; then fixed with GL bioactive isolates by exhaust method.

Organism: MRSA (ATCC 33591); Method: ASTM E2149-10

Activity: 89% (24 hours);

Remarks: Excellent antimicrobial activity

[0085] Example 6:

Cotton fabric treated with tannic acid and bioactive isolates of PY and then combined with GL recorded antibacterial activity of 88% at 24 hours against MRSA

Treatment: Cotton fabric pretreated with tannic acid and fixed with PY bioactive isolate then fixed with GL bioactive isolates by exhaust method

Organism: MRSA (ATCC 33591); Method: ASTM E2149-10

Activity: 88% (24 hours);

Remarks: Excellent antimicrobial activity

[0086] Example 7:

Synthetic dyed cotton fabric treated with bioactive isolates and tested its antimicrobial property against MRSA. Synthetic chemical dye fixed textile cotton fabric, treated with alum and bioactive isolates of GL recorded the antibacterial activity of 81% at 24 hours against MRSA.

Treatment: Synthetic chemicals dyed textile cotton fabric then pretreated with alum and fixed with GL bioactive isolates. Organism: MRSA (ATCC 33591); Method: ASTM E2149-10

Activity: 81% (24 hours);

Remarks: Excellent antimicrobial activity

Table-2: Evaluation of treated textile material using various bioactive isolates against MRSA resistant activity as mentioned in in examples 1- 7.

[0087] The textile materials were prepared by various methods such as pre- mordanted with alum, tannic acid or without alum & tannic acid padded methods, exhausts method and also combination of one or more bioactive extract to obtain antimicrobial activity. The MRSA resistant activity was evaluated by an independent laboratory SITRA, Coimbatore.

then coated with PH bioactive isolates by exhaust

method - second trial

Cotton fabric premordanted with alum and then GLA 88 coated with GL bioactive isolates by exhaust

method

Cotton fabric premordanted with tannic acid and PYT 88 coated with PY bioactive isolates; then again coated +

with GL isolates by exhaust method GL

Cotton fabric premordanted with tannic acid and PHT 89 coated with PH bioactive isolates; then again coated +

with GL isolates by exhaust method GL

Cotton fabric premordanted with alum and coated GLA 98 with GL bioactive isolates by exhaust method (after

three washes)

Cotton fabric premordanted with alum and coated GLA 98 with GL bioactive isolates by exhaust method (after

30 washes)

Cotton fabric premodanted with tannic acid and CuT 56 coated with - Cur bioactive isolates by exhaust

method

Organism: MRSA strain used (ATCC 33591); Method: ASTM E2149- 13a

Cotton fabric coated with Hr bioactive isolates - by HrC 90.8 exhaust method

Cotton fabric pretreated with alum and coated with HrA 94.8 Hr bioactive isolates by exhaust method

Cotton fabric pretreated with tannic acid and coated HrT 99.9 with -Hr bioactive isolates by exhaust method

Synthetic blue Cotton fabric coated with Hr SB HrC 76.8 bioactive isolates by exhaust method

Synthetic blue Cotton fabric pretreated with tannic SB HrT 41.1 acid and coated with Hr bioactive isolates by exhaust method

Synthetic green Cotton fabric coated with Hr SGHrC 53.5 bioactive isolates - by exhaust method

Cotton fabric premodanted with alum and coated GLA 99.96 with GL bioactive isolates by exhaust method

Table 3

[0088] Table 3: The stability of the antimicrobial activity especially against MRSA of the textile material treated with the microbial bioactive isolates stored for a year period.

Evaluation of MRSA resistant activity of textile fabric samples prepared by various methods and extracts

[0089] All the prepared samples have been sent to Sree Chitra Tirunal Institute for Medical Sciences & Technology, (SCTIMST), Trivandrum as per ASTM method and results are provided in the following table 4.

Table 4: Evaluation of bioactive isolates treated textile material for MRSA resistant activity -after one hour

[0090] Under Dynamic Contact Method: The test sample was sterilized by autoclaving before performing the test. The test sample was aseptically transferred in to the medium inoculated with the MRSA strain and incubated for one hour. Enumeration was carried out using plate count method (spread plate technique).

PH bioactive isolates by exhaust method (Porous fabric)

Cotton fabric premordanted with alum and GL bioactive 31 isolate by exhaust method

Table 5: Evaluation of antibacterial textile material especially against MRS A and VRSA (after a year period)

[0091] Experiment carried out at SCTIMST, Trivandrum

Under Dynamic Contact Method: The test sample was sterilized by autoclaving before performing the test. The test sample was aseptically transferred in to the medium inoculated with the MRS A and VRSA strain and incubated for one hour. Enumeration was carried out using plate count method (spread plate technique)

method (CuT)

Cotton fabric premordanted with tannic acid and

coated with PH bioactive isolates (Bandage 68.3 59.2 fabric) - by exhaust method (PHT)

Cotton fabric premordanted with tannic acid and

coated with PH bioactive material (Bandage 80 48.3 fabric) by exhaust method - Second trial (PHT)

Cotton fabric premordanted with alum and coated

with GL bioactive material (Bandage fabric) by 30 46.3 exhaust method (GLA)

Cotton fabric premordanted with tannic acid and

coated with PH isolates; again coated with GL 17 31 isolates by exhaust method (PHT + GL)

Cotton fabric premordanted with tannic acid and

coated with PY isolates; then again coated with 13.9 49 GL isolates by exhaust method (PYT + GL)

Cotton fabric premordanted with alum and coated

with PH bioactive isolates (PHA) by exhaust

56 64 method

Cotton fabric premordanted with tannic acid and

coated with PH bioactive isolates (PHT) by

27 50 exhaust method

Cotton fabric premordanted with alum and coated

with GL bioactive isolates (GLA) by exhaust

26 65 method

Cotton fabric coated with PH bioactive isolates

47 81 (PHC) by exhaust method

Cotton fabric premordanted with tannic acid and

coated with PY bioactive isolates (PYT) by 28 51 exhaust method

Cotton fabric coated with S.parvulus bioactive 51 37 isolates (SP) by exhaust method

Table 6: Evaluation of antibacterial textile material for MRSA, VRSA activity and P. aeruginosa resistant activity (after one and half year period)

[0092] Experiment carried out at SCTIMST, Trivandrum

Under Dynamic Contact Method: The test sample was sterilized by autoclaving before performing the test. The test sample was aseptically transferred in to the medium inoculated with the MRSA and VRSA strain and incubated for one hour. Enumeration was carried out using plate count method (spread plate technique).

Test Method ASTM - E2149-10 & 13a

Sample preparation The test sample was sterilized by autoclaving before performing the test.

The test sample was aseptically transferred in to the medium inoculated with the MRS A and VRSA strain and incubated for one hour.

Methicillin resistant S.aureus(MRSA) ATCC 700698

Specific Microbial strain used

Vancomycin Resistant S. aureus (VRSA)

for the study

Pseudomonas aeruginosa ATCC 27853

Application of the extract - treatment of Final MRSA Final VRSA Final the textile resistant resistant Pseudomonas activity of activity of resistant the textile the textile activity of the (%) (%) textile (%)

Control Fabric (RFD) - Control sample 9 6.3 5.4

Cotton fabric coated with PH bioactive 88.3 13.9 71.6 material (PHC) by exhaust method

Cotton fabric coated with PY bioactive 62 57.6 73 material (PYC) by exhaust method

Cotton fabric coated with GL bioactive 37 40 17.6 isolates (GLC) by exhaust method

Cotton fabric coated with Hr bioactive 51 54 37 isolates (HrC) by exhaust method

Cotton fabric coated with Curvularia 25 52 47.6 bioactive isolates (CuC) by exhaust

method

Cotton fabric premordanted with tannic 53 40 52 acid and coated with PH bioactive isolates (PHT) by exhaust method

Cotton fabric premordanted with tannic 78 96 86.3 acid and coated with PY bioactive isolates

(PYT) by exhaust method

Cotton fabric premordanted with alum and 13.6 61.5 14.27 coated with GL bioactive isolates (GLA)

by exhaust method

Cotton fabric premordanted with tannic 39.5 74.3 10.9 acid and coated with Hr bioactive isolates

(HrT) by exhaust method

Cotton fabric premordanted with tannic 41.6 75 38 acid and coated with Cur bioactive

isolates (CuT) by exhaust method

Advantages of the present disclosure

[0093] Bioactive isolate coated in the textile material exhibited consistent antibacterial activity against MRSA, VRSA, Pseudomonas aeruginosa, Vancomycin Resistant Enterococci, Klebsiella pneumoniae, Streptococcus pneumoniae, Salmonella sp., Escherichia coli, extensively drug resistant tuberculosis, Acinetobacter baumannii, Neisseria gonorrhaeae, Clostridium difficile, and Candida sp. related infections.

[0094] The treated textile material exhibit antimicrobial activity even after at multiple washings. Easy method of application on fabrics. No deterioration of fabric quality.

[0095] The biological material such as microfungi, macrofungi etc. may be grown in short duration with simple cultivation / production techniques. [0096] The process of treating the textile material or the treated textile product does not produce any harmful effects such as skin irritation to the manufacturer, user and the environment.

[0097] Single or in combination of different antimicrobial metabolites recorded enhanced antibacterial activity against MRSA and VRSA.

[0098] Synthetic / chemical dyed cotton fabric coated with the bioactive isolates recorded the excellent antibacterial activity against MRSA.

[0099] Cost effective, environment friendly and reproducibility.

[00100] The treated textile (Meditex) product satisfies the irritation studies at Textile and Medical Research Laboratories and recorded the performance.