Recombinant secretory proteins based new ‘Cocktail ELISA’ as a marker assay to differentiate infected and vaccinated cows for Mycobacterium avium subspecies paratuberculosis infection

Johne's disease is endemic in the domestic livestock population of India. Recently, we developed highly effective 'Indigenous vaccine' to control Johne’s disease in animals. In order to gain disease free status as per World Organization for Animal Health, it is essential to have a marker assay to differentiate between infected and vaccinated animals before vaccine can be used in the field. We have developed a new marker assay ‘Cocktail ELISA’ using six ‘recombinant secretary proteins’ (MAP 1693c, MAP 2168c, MAP Mod D, MAP 85c, MAP Pep AN and MAP Pep AC) and evaluated for diagnosis of Johne’s disease along with 'Indigenous ELISA kit'. This ‘Cocktail ELISA’ successfully differentiated the infected, vaccinated and healthy (non-infected) cows and will facilitate the use of Johne’s disease vaccine to control the disease in cows at national levels.

‘Indigenous’ and ‘Ethanol Vortex’ ELISA kits for diagnosis of Mycobacterium avium subsp. paratuberculosis infection in cattle: Is there a ‘globally relevant kit’ in the ‘Reverse Ice-burg’ environment?

Highly versatile and robust 'Indigenous ELISA kit' for the diagnosis of Mycobacterium avium subsp. paratuberculosis infection in cattle herds was compared with 'Ethanol Vortex (EV) ELISA kit' of USA. Of 160 (118 vaccinated and 42 non-vaccinated) cattle screened, 129 and 35 were positive in 'Indigenous' and 'Ethanol Vortex' ELISA kits, respectively. 'I-ELISA', using 'semi-purified protoplasmic antigen' from native highly prevalent biotype ('Indian Bison type') of MAP of goat origin, was highly sensitivitive (91.4%) as compared to the 'EV-ELISA. 'I-ELISA kit using whole cell sonicate from native 'S 5' (‘Indian Bison type’) strain of MAP as 'antigen source' was significantly superior than EV-ELISA kit using surface antigens from 'Linda' strain ('cattle type') of cattle origin in USA. Therefore, 'i-ELISA kit' may be recommended for the screening of domestic cattle herds against MAP infection in India. The present study has demonstrated that in the diagnosis of chronic infections and diseases, such as Johne's disease, 'Indigenous kits' are significantly superior to kits made in other countries, EV-ELISA, in the present case, particularly screening of native cattle herds endemically infected with MAP.

Wound healing potential of Ocimum sanctum Linn. with induction of tumor necrosis factor-α.

Ocimum sanctum, a well known herb in Indian medicine, possesses various therapeutic properties including healing properties and cytokine induction. Wound healing activity of cold aqueous extract of O. sanctum leaves along with its effect on tumor necrosis factor- (TNF-α) was assessed using excision model of wound repair in Wistar albino rats. After application of the O. sanctum extract, rate of epithelization with an increase in wound contraction was observed. In animals, treated with 10% O. sanctum extract in petroleum jelly, wound healing was faster as compared to control group which were treated with petroleum jelly alone but significant accelerated healing was noticed in animals which in addition to the topical application of 10% extract of O. sanctum, were prefed with 250mg/kg body weight of aqueous O. sanctum extract daily for 20 consecutive days. During wound healing phase TNF-α level was found to be up regulated by O. sanctum treatment. Early wound healing may be pronounced due to O. sanctum extract, by elevating TNF-α production.