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A DBT-TANUVAS Partnership Programme
TECHNOLOGIES - COMMERCIALIZED
QUICK HEAL – TOPICAL WOUND HEALING CREAM

A Collagen based curcumin gel as a topical cream has been developed that enables quick wound healing. The technology for this product was transferred to M/s Sihil Pharma companion health division, Chennai.

Brief Description:

Quick heal is specially formulated wound healing gel prepared from animal collagen enriched with curcumin and antibiotics that helps the biological mechanism of wound healing with external collagenous connective tissues in a special gel form which merge with a regenerative connective tissues in the wound surface and makes the healing process faster it also helps in reducing the scar lesion formation in the wound area.

Quick heal can be used for all cuts, wounds (both infected and non infected) anal fistula, ulcer, maggots, FMD diseases, Footrot lesions in sheeps, teat and tail injuries heals faster without scar formation.

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TECHNOLOGIES - AVAILABLE FOR COMMERCIALIZATION
ABT CHOICE – VALIDATION OF THE MAGNETIC PARTICLE WITH CLINICAL MASTITIS ISOLATES
Need for the project:

Nanoparticles are applied for enrichment of target cells from complex matrix and thereby improve the efficiency of detection techniques. In an earlier study from our group the efficacy of a glycine coated magnetic nanoparticle to entrap standard strains of mastitis causing bacteria from milk samples and used it to identify the sensitivity of antibiotics for clinical application (Viswanathan et al., 2014). We have determined the efficacy of the particle in trapping the clinical isolates and also the sensitivity of PCR in detecting the multitude of pathogens and some of their resistance genes in a single tube assay. The nanoparticle not only enabled sensitive detection, but also enabled identification of drug resistant bacterial strains like Methicillin Resistant Staphylococcus aureus (MRSA).

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Objective:
  • To generate additional data generation with the respect to the binding efficiency of the magnetic nanoparticles to different E. coli, Staphylococci sp. and Streptococci sp. isolates from mastitis cases
Outcome:
  • Efficacy of the nanoparticles to trap various mastitis pathogens from complex matrix like milk samples
Current status:
  • Preliminary data generated on the binding efficacy of the magnetic particle to few isolates
  • The detection efficiency of PCR is 17 CFU/ml for E.coli and 50 CFU/ml for Staphylococcus aureus
Large scale validation needed for which DBT funding has been sought through Project Under Nanotechnology task force.
ANTIMICROBIAL EGG TRAY
Objective:

In recent years, the egg and egg product contaminations, associated out breaks particularly by bacterial species were reported worldwide. In developing countries, reusable egg trays are used without recycling for storing and selling eggs. Several studies have reported a higher incidence of bacterial contamination occurring through the egg trays in retail outlets than in the eggs themselves. The shelf life of the egg is directly influenced by egg shell's quality.

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Brief description:

The success of the research work including the fabrication of the egg tray, studies their anti-microbial activity, and their applications. To develop the egg tray, a hybrid mixture was prepared based on chitosan and gelatin. Later the silver nanoparticles were embedded on it. The above hybrid mix was coated on paper through manual spray coating. This hybrid mixture coated egg trays were used to store table and fresh farm eggs.

Performance indicator:
  • The series of studies needed to confirm the hybrid mixture coating (SEM, TEM, XRD, FTIR, EDX and XPS).
  • The egg quality (8 different parameters recommended by ISI standard) and the microbial load on the egg shell surface was examined and compared with control samples.
Current status:
  • 100 antimicrobial egg trays were already made in TRPVB and their utility was tested with form, table eggs.
  • The experimental data was documented and the final project report is under progress.
Commercialization effect:
Egg tray manufacture from Namakkal has shown interest.
NANO NDV
Objective:

Newcastle disease virus (NDV) is an avian virus that causes disease in more than 250 avian species. In the poultry industry, it causes major economic losses.

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Brief description:

The success of the research work including the fabrication of the NDV conjugated nanoparticles, studies their vaccine delivery efficiency, and the immunological responses. To develop the Nano NDV, the CaP NPs was prepared based on -Cyclodextrin and calcium chloride. Later the nano particles were functionalized with APTES. The amine functionalized nanoparticles were mixed with NDV virus and delivered through oculonasal on chicken.

Performance indicator:
  • The series of studies needed to confirm the NDV conjugation on CaP NPs (SEM, TEM, XRD, FTIR, EDX, Raman and PCR).
  • The vaccine delivery on chicken was monitored based on HI titre, ELISA and NDV gene expressions and the immunological responses were compared with commercial vaccine.
Outcome:
  • The Nano NDV showed improved vaccine delivery and shelf life upon longer storage in room temperature.
Current status:
  • 1000 Nano NDV vaccine vials were already made in TRPVB and their utility was tested in farms.
  • The experimental data was documented and the final project report was already prepared.
Commercialization effect:
Hester Bio Sciences (P) Ltd showed interest and collaborative project under consideration by DBT.
H2O2 DETECTION KIT FOR MILK, HUMAN, CONSUMER PRODUCT AND VETERINARY CLINICAL APPLICATIONS
Need for the project:

In the milk industries, the instruments used for the milk sample cooling, mixing, transporting, bottling and packing were cleaned twice in a week with H2O2 as a contaminant. The H2O2 is also used as stabilizer in milk, in well-defined concentrations, such as the preparations of hard cheeses as an alternative to pasteurization.

In Human, the H2O2 is generated by various oxidase-catalyzed reactions; a highly sensitive determination method of H2O2 is applicable to measurements of low levels of various oxidases and their substrates such as glucose, lactate, glutamate, urate, xanthine, choline, cholesterol and NADPH. More ever the consumer product, such as the hair dyes contained large amount of hydrogen peroxide. So the sensitive detection of hydrogen peroxide (H2O2) plays an important role.

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Brief description:

The success of the research work including the fabrication of the fluorescent nanoprobe, studies their specificity, sensitivity, stability and their applications. To develop the nanosenosors, a fluorescent nanoparticles was prepared based on rhodamineisocyanide and calcium phosphate. Later the Rho/CaP nanoparticles were functionalized with HRP. The functionalized nanoparticles were incubated with H2O2 in the presence of OPD. The end product was read at three different methods such as fluorometeric, colorimetric and visual detections.

Analytical advantages of this kit:
  • Ability to be monitored by the naked eye
  • Three different readout system possible
  • Simple assay steps
  • Highly suitable for H2O2 detection in bacterial culture medium
  • Naked eye detection method was designed to make the assay field-compatible.
Performance indicator:
  • The series of studies needed to confirm the HRP conjugation on Rho/CaP NPs (SEM, TEM, XRD, FTIR, EDX, Raman and Spectrofluorometer analysis).
  • The probe stability, detection sensitivity, specificity and the real sample analysis need to perform.
Outcome:
  • The HRP/Rho/CaP nanoparticles sensors showed the ultrahigh sensitivity, excellent stability and enlarged linear range of detection of hydrogen peroxide. The fluorometric and colorimetric performance was comparable to a commercial method.
Current status:
  • The portable kits were already made in TRPVB and their utility was tested with milk, bacterial samples.
  • The experimental data was documented and the final project report is under progress.
Commercialization effect:
The total milk quality test kit design under progress along with this.
DEVELOPMENT OF ORAL/SKIN PROBIOTIC SPRAY FOR DOGS – ORO-CUTE®
Brief Description:

Probiotic organisms are friendly organisms that can inhibit the pathogenic organisms by competitive inhibition, maintaining lower pH, production of bacteriocins etc., and has immunomodulation property. Probiotics are being used in various veterinary and human applications. TRPVB has undertaken development of Probiotic spray for dogs to maintain healthy oral cavity and skin.

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Twenty two Lactic Acid Bacteria were isolated from oral/skin/ aural samples of healthy pups. They are identified up to species level by 16S rRNA sequencing and biochemical tests. The isolates were checked for their Probiotic characters by checking the attachment to dog buccal epithelial cells, antimicrobial activity against Staphylococci, E. coli, Hydrogen peroxide release, Antibiotic sensitivity etc. Depending on these characters 5 isolates namely Lactobacillus plantarum, Weissellacibari, Pediococcuspentosaceus, Enterococcus munditii, Enterococcus faecium were selected. The isolates were scaled up using fermenters and spray dried at Microbax India Ltd., Hyderabad. A probiotic formulation using consortium of these five organisms with varied counts per dose was formulated. Stability studies for spray dried powders indicated that when stored in 40⁰C survival is 100% till 12 months. After reconstitution, L. plantarum and W. cibaria were maintained for 30 days at 40⁰C whereas the Enterococcus/ Pediococcus were maintained for 60 days.

Clinical trials in dogs were conducted to check the efficacy and dose response of the formulation. Results of the study indicated that the spray is effective in reducing the halitosis, dental tartar, reducing CFU of aerobic and anaerobic bacteria by effective colonization of LAB. Daily spray for 4 days and once in 5 days for 4 times is effective for the above parameters of study. The effect was maintained till the study period, i.e 30 days after withdrawal of the spray.

Objectives
  • Developing Probiotic spray for dog to maintain healthy oral cavity and skin using Lactic Acid Bacteria (LAB) isolated from oral/aural/skin of healthy pups.
  • Up scaling and spray drying the Probiotic isolates and formulating an oral/skin spray and packaging it.
  • Validation of the product by conducting clinical trials and commercialization.
Current status:
  • On development and validation
  • Study document prepared
  • Product Development: Completed
  • Patent filing: Pending with attorney
  • Commercial packaging: Completed
Commercialization efforts:
Talks with Microbax India ltd., did not materialize. Other partners need to be identified M/s. Genomix Biotech, Hyderabad has enriched interest.
Recombinant ESAT-CFP10 Protein
Brief Description:

Bovine tuberculosis (BTB) is a major animal health problem and the approved BTB diagnostic tests are single intradermal tuberculin test (SITT) using purified protein derivative (PPD) of bovine tuberculin and/or in vitro assays to measure BTB specific interferon gamma (IFN-γ).

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Back drop:
Though the SITT is widely used in India, this diagnostic method suffers from several disadvantages.
  • PPD lacks specificity because it is an undefined crude extract of water soluble proteins from a heat-treated culture of M. bovis.
  • The specificity of the tests is compromised due to its inability to distinguish the infection of pathogenic mycobacterium from the non-pathogenic environmental mycobacterium.
Technology solution:
To overcome the above problems recombinant TB antigens have been used instead of PPD.
  • Early secretory antigenic target 6 (ESAT-6) and culture filtrate protein 10 (CFP-10) are encoded by genes of the region of difference -1 (RD1) of M. bovis and are immunologically dominant proteins in TB infected cattle.
  • ESAT-6 and CFP-10 are particularly robust inducers of recall IFN-γ responses in infected cattle. Genes for these proteins are absent in many environmental, non-tuberculosis mycobacterium as well as in the BCG vaccine.
  • The assays using these proteins can be applied to differentiate infected animals from vaccinated animals (DIVA) and it was proven that peptide cocktail derived from the mycobacterial antigens ESAT-6, CFP-10 and Rv3615c allowed differentiation between BTB infected and BCG vaccinated cattle when used as a skin test reagent
  • The fusion protein ESAT-6:CFP-10 could effectively replace the individual proteins ESAT-6 and cocktail.
Available TB diagnostic antigens:

TRPVB offers E.coli expressed TB specific Fusion protein of ESAT-6: CFP-10 (~30kDa) for research and diagnostic purposes.

Application
  • For in vitro assays to measure TB specific IFN-γ.
  • For in vivo assay such as SITT, DIVA etc
  • Research application
Adventitious Virus Detection
Back drop:

The recent reports on the presence of extraneous viruses such as porcine circovirus and porcine parvovirus contamination in rotavirus vaccine and the identification of nodovirus in the insect cells which was used to produce human papilloma virus (HPV) vaccine had cautioned the regulators and vaccine industry about the need for extensive characterization of the vaccine banks and substrates.

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The major sources of extraneous Viruses contamination in vaccine and cell banks are animal derived raw materials such as porcine trypsin, bovine serum, cell line, source of Virus isolation, etc used during the vaccine development and production process.

Currently the 9CFR 113 extraneous Virus testing protocol was practiced by vaccine industry to screen their Virus and cell banks. However, there is an extensive list of bovine, porcine and human Viruses which are not detected by laborious and time-consuming 9CFR 113 protocol.

Regulators and manufacturers are in great need of additional testing procedures which may be useful as adjuncts and replacements for the 9 CFR tests in quality control labs of vaccine manufacturers.

Technology available:
TRPVB has developed a multiplex PCR assay for detection of adventitious Viruses of porcine and bovine origin.
  • TRPVB has developed multiplex PCR assay for screening FBS for five prominent bovine Viruses and for screening porcine trypsin for six prominent porcine Viruses
  • Initial screening of veterinary vaccines (n=17), human vaccines (n=10), and trypsin (n=7) at TRPVB indicated that 52% of veterinary vaccines and 10% of human vaccines and 100% of trypsin showed adventitious viral genome contamination.
Application of this technology:
This assay would be useful for,
  • Vaccine manufacturers for screening animal derived raw materials used in vaccine manufacturing
  • Vaccine QC testing labs
  • R&D laboratories and research institutes
  • Animal disease diagnosis laboratories
  • Academic institutions
Wild TB Alert Kit
INTENDED USE:

The Tuberculosis (TB) Antibody Rapid Detection kit is a qualitative, single use and two-step immuno chromatographic screening test for the detection of antibodies to Mycobacterium tuberculosis complex in serum, whole blood and plasma from wild animals. It is intended to be used as a preliminary screening test and as an aid in the diagnosis of infection with tuberculosis in wild animals. Any reactive specimen with the Genomix Tuberculosis Antibody Detection Test must be confirmed with alternative testing method(s).

Download Wild TB Alert Kit Brochure
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Last updated on 18 Feb 2017