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Abstract
7. SUMMARY
(1) This study comprised the examination of 420 tracheal and sinus swabs collected from turkey flocks in two Governorates (Gharbia and El-Wadi El-Gedid) for detection of Mycoplasma infection. It was found that, isolation percent from tracheal and sinus swabs in Gharbia Governorate were 8.89% and 15.56%, respectively and in El-Wadi El-Gedid Governorate were 31.67% and 35.83%, respectively.
(2) The highest incidence of Mycoplasma isolation was obtained from sinus swabs in El-Wadi El-Gedid Governorate (35.83%) while the lowest recovery rate was obtained from tracheal swabs in Ghabia Governorate (8.89%).
(3) The rate of Mycoplasma isolation from sinus swabs was higher than tracheal swabs in both Governorates.
(4) Biochemical characterization and serological identification proved that 95 isolates were M. gallisepticum, 3 isolates were M. synoviae and 5 isolates were M. meleagridis.
(5) One hundred eighty serum samples were also collected from turkey flocks in both Governorates and were examined by SPA and ELISA tests for detection of antibodies against M. gallisepticum, M. synoviae and M. meleagridis.
Results of SPA and ELISA indicated that all isolates were M. gallisepticum, M. synoviae and M. meleagridis.
In El-Wadi El-Gedid Governorate, SPA results showed 42 positive samples out of 90 for M. gallisepticum and no positive samples for M. synoviae. While results of ELISA showed 19 positive samples and 55 suspected samples out of 90 for
M. gallisepticum (with GMT 1044.1), no positive samples for
M. synoviae and 47 positive samples and 23 suspected samples out of 90 for M.meleagridis (with GMT 1389.7).
While in Gharbia Governorate, out of 90 serum samples SPA results showed 15 positive samples for M. gallisepticum and 8 positive samples for M. synoviae. While results of ELISA showed 4 positive samples and 18 suspected samples for M. gallesepticum (with GMT 899.7), 6 positive samples and 4 suspected samples out of 90 for M. synoviae (with GMT 1951) and 8 positive samples and 14 suspected samples out of 90 for
M. meleagridis (with GMT 1266.9)
(6) SDS-PAGE was used as confirmatory test for identification of M. gallisepticum (MG) and M. synoviae (MS) field isolates. The electrophoretic profile of MG reference strain (PG31) revealed 27 protein bands extended from 16.27 to 242.04 KDa. Different field isolates gave 26 to 30 bands ranged from 15.87 to 244.94 KDa. Most similar bands lay in the area between 161.71 to 242.09, 29.9 to 77.94 and 16.27 to 24.26 KDa. Field isolates characterized by presence of extra 9 bands which didn’t present in the protein profile of the reference strain. While the electrophoretic pattern of MS reference strain gave 21 protein bands ranged from 16.14 to 240.11 KDa and MS field isolates produced 24 to 30 bands ranged from 16.14 to 240.98 KDa. Field isolates almost contain all protein bands in the reference strain but they also had extra bands at 17.38, 19.26, 19.53, 26, 58.94, 74.14, 85, 90.12, 139.54, 209.69 and 227.06 KDa.
(7) Five field isolates were tested by PCR and compared with MG reference strain (PG31 strain). All the examined field isolates were identified as M. gallisepticum (gave characteristic 330 bp fragment). While 2 field isolates were tested and compared with MS reference strain (WVU 1853). Field isolates were identified as M. synoviae (gave characteristic 1100 bp fragment).
(8) Arbitrary –primed PCR (AP-PCR) is a more recent technique used for differentiation among different strains of the same Mycoplasma species. Also, it is a reproducible method for comparing the Mycoplasma field isolates in epidemiological studies. The technique detects the genetic diversity in natural populations among field isolates. DNA profiles of 4 MG field isolates were compared with that of MG reference strains (F, PG31, R) using Fan 1 and Fan 2 primers. The banding patterns of 4 field isolates were highly similar to those of MG reference strains. Also, DNA profile of MS field isolate were compared with that of MS reference strain. DNA profile of reference strain was similar to that of the field isolate. The field isolate shows 3 common bands with MS reference strain.