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Background Clostridium perfringens , a common environmental bacterium, is responsible for a variety of serious illnesses including food poisoning, digestive disorders, and soft tissue infections. Mastitis in lactating cattle and sudden death losses in baby calves are major problems for producers raising calves on dairy farms. The pathogenicity of this bacterium is largely mediated by its production of various toxins. Results The study revealed that Among the examined lactating animals with a history of mastitis, diarrheal baby calves, and acute sudden death cases in calves, C. perfringens was isolated in 23.5% (93/395) of the total tested samples. Eighteen isolates were obtained from mastitic milk, 59 from rectal swabs, and 16 from the intestinal contents of dead calves. Most of the recovered C. perfringens isolates (95.6%) were identified as type A by molecular toxinotyping, except for four isolates from sudden death cases (type C). Notably, C. perfringens was recovered in 100% of sudden death cases compared with 32.9% of rectal swabs and 9% of milk samples. This study analyzed the phylogeny of C. perfringens using the plc region and identified the plc region in five Egyptian bovine isolates (milk and fecal origins). Importantly, this finding expands the known data on C. perfringens phospholipase C beyond reference strains in GenBank from various animal and environmental sources. Conclusion Phylogenetic analyses of nucleotide sequence data differentiated between strains of different origins. The plc sequences of Egyptian C. perfringens strains acquired in the present study differed from those reported globally and constituted a distinct genetic ancestor.

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.

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.

Avian mycoplasmas were mainly the cause of poultry industry economic losses; reduced meat and egg production and increases the antibiotic treatment cost. Mycoplasma gallisepticum (MG) infection is designated as infectious sinusitis of turkeys and chronic respiratory disease of chickens (gasping, depression, semi closed eyes, infraorbital sinuses edema and decrease in egg production). This study aimed to prepare, evaluate and Compare in-house ELISA kits and lateral flow assay (LFA) from a local strain of MG with commercial ELISA kits and PCR consequently. A total of 54 samples (27 tracheal swabs, 10 trachea and 17 lung) and 50 serum samples collected from birds suffering from chronic respiratory disease were tested by prepared in-house ELISA, commercial ELISA kits, PCR and LFA; a high correlation coefficient between in-house ELISA using whole antigen or sonicated antigen and commercial kit was recorded. Lateral Flow assay (LFA) performance indicate a low sensitivity (77.5%) but maintain a high specificity (92%) compared to PCR. The in-house ELISA kits and LFA prepared could be used as a fast diagnostic technique for detection of MG in Egypt. According to the available knowledge the prepared LFA for diagnosis of MG infection in chickens was developed for the first time in Egypt.

Rabies is a severe viral infection that causes acute encephalomyelitis, with a case fatality rate of nearly 100% following the onset of neurological clinical signs. Rabies irreversible clinical signs development can be effectively avoided with post-exposure prophylaxis (PEP), which includes vaccines and anti-rabies immunoglobulins (RIGs); however, there is no treatment for symptomatic rabies. The major PEP protocol faces serious access and implementation obstacles in association with a resource-limited setting, which could be successfully overcome by substituting RIGs for monoclonal antibodies (mAbs). Lower production costs, consistent supply availability, long-term storage/stability, and an improved safety profile are all advantages of mAbs. The current work focuses on the key characteristics of currently developed mAbs against rabies and highlights their potential as a novel therapeutic approach. Using immunizing Freund adjuvanted emulsions of inactivated purified Vero cell rabies vaccine (PVRV, VERORAB) produced by Aventis Pasteur to immunize the BALB/c mice. The immunized BALB/c mice were tested for the production of anti-rabies virus-specific antibodies using Enzyme-linked immunosorbent assay (ELISA). High-responder mice were selected for the fusion process. Hybridomas recovered from the fusion process were selected and separated from the unfused cells and unfavorable fused cells by using the selective HAT medium. Twelve days post fusion the produced hybrids were screened for production of Rabies virus-specific antibodies using ELISA. Four murine hybridomas secreting rabies virus-specific monoclonal antibodies (mAbs) have been properly developed. These 4 stable hybrids were successfully cloned into 4 stable clones, namely, 1E4, 1E9, 2F3, 4E1. The rabies virus specific monoclonal antibodies produced by the 4 selected hybridomas were of IgM isotype. Using Western Blot technique, the specificity of the produced hybrids was confirmed. The neutralizing potential of the prepared mAbs was evaluated and the efficacy of mAbs cocktail prepared from the 4 hybridomas to protect mice in post exposure therapy was determined. The mAbs cocktail given to mice at 24 hours post infection was able to offer 100% protection to mice challenged with 1000 LD50 of rabies virus strain whereas all control mice developed the disease.Competing Interest StatementThe authors have declared no competing interest.

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%.