Ringworm is a worldwide distributed contagious disease infecting both man and animals that constitute an economic, zoonotic, and health problem concern all over the world. During the last decade, attention has been directed to vaccination as an ideal approach to the control of such diseases. In the present study, non-adjuvanted polyvalent vaccines were prepared from locally isolated hot and virulent dermatophyte species, namely Trichophyton verrucosum (T. verrucosum), Trichophyton mentagrophytes (T. mentagrophytes), and Microsporum canis (M. canis) were immunologically evaluated. The prepared vaccine evaluation was focused on the aspects of immunogenicity and protective efficacy using guinea pigs. Both in its living or inactivated forms, the vaccine-induced significant humoral and cell-mediated immune responses and achieve proper protection of guinea pigs against challenging infections with homologous and heterologous dermatophyte strains. On the other hand, investigations on dermatophyte exo-keratinases showed that it was better produced and more expressed in a mineral-based medium containing pure keratin (3 g/L) than in the same medium with human hair supplementation (2.6 g/L). The maximum dermatophyte productivity of exo-keratinases was found to be between 18 and 21 days post-incubation. Using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), two fractions with molecular weights of 40 kDa (fraction I) and 28 kDa (fraction II) have been identified in the culture filtrate of the three involved dermatophyte species. Both fractions demonstrated keratinolytic activity. The specific activity of the isolated keratinases (number of Keratinase units (KU)/mg protein) was stronger in fraction I, where it reached 18.75, 15.38, and 14 KU/mg protein as compared to 12.9, 8.74, and 12 KU/mg protein in fraction II of T. verrucosum, T. mentagrophytes, and M. canis, respectively. The dermatophyte exo-keratinases proved to be immunogenic as they stimulated high keratinase-specific antibody titers and induced strong delayed skin hypersensitivity reactions in vaccinated animals. Anti-keratinase-specific IgG was detected in sera of guinea pigs immunized with the inactivated or living polyvalent dermatophyte vaccines by a homemade enzyme-linked immunosorbent assay (ELISA) using dermatophyte exo-keratinases as coating antigen. The intradermal injection of dermatophyte exo-keratinases induced specific delayed skin reactions in guinea pigs immunized with the inactivated or the living polyvalent dermatophyte vaccines. The intradermal injection of dermatophyte exo-keratinases in the control non-sensitized guinea pigs was associated with itching, swelling, and bloody scar formation, however, no skin indurations were formed. The development of those post-exo-keratinases injection reactions in the control non-sensitized apparently healthy guinea pigs group, suggests an exo-keratinases possible role in the pathogenesis of dermatophytosis.
The intradermal tuberculin skin test has been in use for almost a century in the diagnosis of bovine tuberculosis
Dermatophytosis is a widely spread contagious zoonotic disease, affecting both man (tinea) and animals (ringworm). This disease is caused by a group of closely related keratinophilic fungi known collectively as the dermatophytes group. Although the wide distribution of dermatophytosis cases throughout the whole world and its adverse clinical effect on human health, economical effect on productive animals, and pet animal welfare, there is no rapid accurate diagnostic tool for such disease. The current conducted study tries to accomplish the difficult equation by achieving an accurate, sensitive, specific, user-friendly, rapid, robust, device-less, deliverable to end-users, and economic cost for the development and production of diagnostic kits. Through the development of a rapid diagnostic kit based on immunochromatographic assay with three major affordable reproducible production stages; preliminary stage, developmental and standardization stage, and evaluation stage. Obtaining dermatophytes-specific polyclonal antibodies against criteria-based selected dermatophytes strains associating proper gold nanoparticle preparation, characterization, and conjugation, with proper loading of the different bio-reactants on the efficiently laminated and fabricated lateral flow strips were the main challenge and control points through the whole process. Also, as a result of examining 100 animal samples using the new kit, the κ coefficients of the kit with the direct microscopy while the kit with the culture were 0.44 and 0.76, respectively. Therefore, the newly designated and developed kit showed a very promising competitive diagnostic result within 5–7 min through easy-to-be-performed three steps.
Poultry, mainly chickens and its white meat represents one of the main, nutritionally valuable, and affordable red meat replacer source of protein throughout the whole world with special reference to developing countries. A long list of microbial agents especially bacterial pathogens threat chickens production cycles. They constitute one of the major problems facing the rapidly expanding poultry industry and are responsible for considerable economic losses. ). Fowl Cholera, Infectious Coryza, and Ornithobacterium Rhinotracheale (ORT) diseases were among the serious bacterial infections that affect respiratory tract of chickens with an global adverse effect on poultry production. A formalized whole culture vaccine of composed of P. multocida serotypes A5, A8, A9 and D2, Avibacterium paragallinarum serotype A and C and Ornithobacterium rhinotracheale serotype A was prepared. This polyvalent vaccine proved to be safe producing no adverse side effects when injected in chickens. The immunizing efficacy of this vaccine was evaluated in SPF chickens, which were immunized at 6 weeks of age. The protective efficacy of the vaccine was determined using challenge test. The developed vaccine was effective in protecting chickens against Fowl Cholera, Infectious Coryza and Ornithobacterium Infection (ORT diseases) in chickens against challenge with these pathogens. Vaccinated chickens challenged with virulent Pasteurella Multocida serotypes A5, A8, A9 and D2 showed protection rates of 86.6%, 93.3%, 93.3% and 93.3%, respectively, as compared with 100% mortality in the non-vaccinated control. Vaccinated chickens challenged with Avibacterium paragallinarum serotypes A and C showed protection rates of 86.6% and 93.3%, respectively. Also, the protection rate against challenge with virulent Ornithobacterium rhinotracheale serotype A reached to 96.6%.
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