Amphistomes are snail-borne trematodes infect rumens and reticulums causing acute and chronic diseases in cattle and buffaloes. The economic losses caused by Amphistomes resulted from drop in milk and meat yield in addition to mortalities. Eighty four serum samples (50 from cattle and 34 from buffaloes, 30 from amphistomes infected and 54 from amphistomes free cattle and buffaloes) collected from Giza and Garbia governourates, Egypt. The collected Amphistomes were morphologically and histologically classified. The identified worms of Paramphistomum and Carmyerius were used for the preparation of somatic antigens separately. The collected 84 serum sample were tested twice by indirect ELISA, firstly with Paramphistomum somatic antigen and secondly with Carmyerius somatic antigen. The antigenic profiles of adult Paramphistomum spp somatic antigen and Carmyerius gergaerius somatic antigen were analysed by (SDS-PAGE). Four male Rabbit were used for the preparation of hyper-immune serum against Paramphistomum and Carmyerius somatic antigens. Nine serum samples (two rabbit hyper immune sera; one for Paramphistomum and the another for Carmyerius , 3 serum sample from Paramphistomum infected cattle and 3 serum samples from Carmyerius infected cattle and one serum sample from non-infected cattle as negative control) were blotted and tested twice at the same time on the nitrocellulose membrane by Western blotting techniques., firstly by using Paramphistomum somatic antigen and secondly with Carmyerius antigen. The results of both ELISA and Western blotting were statistically analysed. The sensitivity, specificity and accuracy for ELISA and Western blotting were (74% and 100%),(82.4% and 33.3%) and (79.76% and77.78%) respectively . The antigenic profile of adult Paramphistomum somatic antigen showed 14 distinct protein bands of protein molecular weights ranging from11.5 to 174 KDa. While Carmyerius somatic antigen showed 13 distinct protein bands ranging from 11.5 to 166KDa. One distinct immunogenic band at 63 KDa was found to react with all sera from infected cattle and buffaloes with Paramphistomum somatic antigen while the same serum samples gave one distinct immunogenic band at 71 KDa with Carmyerius somatic antigen. It is concluded that ELISA is more reliable test for early diagnosis of amphistomiasis. There is a strong cross immune reaction between Paramphistomum and Carmyerius.
Background and Aim: Different species of Mycoplasma are associated with many pathological problems in small ruminants including respiratory manifestation, this problem results in significant losses, especially in African countries. This study aimed to (I) study some epidemiological aspects of Mycoplasma species infections in Egyptian sheep and goats at Giza Governorate, (II) diagnosis of Mycoplasma species affections using bacterial isolation and identification, (III) apply the polymerase chain reaction (PCR) for typing of different Mycoplasma species, and (IV) illustrate the phylogenetic tree for the isolated Mycoplasma species and other species from GenBank using the purified PCR product. Materials and Methods: A total of 335 samples were collected from sheep and goats from Giza Governorate in Egypt as 142 nasal swabs from clinically affected animals, 167 pneumonic lungs, 18 samples from tracheal bifurcation, and 8 samples by bronchial wash were cultured on pleuropneumonia-like organisms (PPLOs) media for cultivation of Mycoplasma species. PCR and sequencing and phylogenetic analysis were adopted to identify and classify the isolated Mycoplasma species. Results: A total of 24 Mycoplasma isolates were isolated on PPLO media, identified by biochemical tests, and confirmed and typed by PCR using specific primers. 10 isolates were confirmed as Mycoplasma arginini, four isolates as Mycoplasma ovipneumoniae by PCR, and 10 isolates as undifferentiated Mycoplasma species. A purified isolate of M. arginini and M. ovipneumoniae was sequenced and phylogenetic analysis was illustrated. Conclusion: M. arginini and M. ovipneumoniae are prevalent in Egyptian sheep and goats. Further studies on M. arginini are required due to its high frequency of isolation from pneumonic sheep and goats and also from animals suffer from different respiratory manifestations.
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