الفهرس 
Cover Page Title in EnglishOnly 14 pages are availabe for public view
 |  | from | 32 |  |  |
| | | from | 32 | | |
Abstract
The current study aimed to: 1- Identify the possibility of genetic variation among Mycoplasma synoviae isolated from respiratory and arthritis samples from infected bird 2- Studying other bacteria causing arthritis in chickens and detection of some virulence genes of isolated bacteria. Material and method: 1- A total of 160 chicken samples (100 respiratory samples ,60 joint samples) were collected from birds from different flocks showing respiratory manifestation and arthritis for isolation of M .synoviae 2-A total of 64 samples (9 Tracheal samples and 55 Joint samples)a for detection of M.synoviae direct from tissue . 3- A total of 20 blood samples were collected for detection of antibodies of M.synoviae By ELISA . 4- A total 40 other samples were collected for isolation of other bacteria causing arthritis. Result of this study: 1- For isolation of M .synoviae 160 samples were cultivated on Frey’s broth medium , while 64 samples were identified for M.synoviae by PCR direct on tissue using oligonucleotide primers complementary to the single-copy conserved 5´ end of vlhA gene, amplicons of 350~400 bp. PCR amplification results revealed that six isolates out of 19 suspected M.synoviae from culture were positive for M.synoviae and 26 out of 64 tissue samples were positive for M. synoviae. 2-High-resolution melting (HRM) curve analysis of amplicons using SYPER green fluorescent dye of 10 selected isolates (6 respiratory isolates and 4 arthirtic isolates ) showed that different Melting temperature curves were ranged between 84.6-85.1°C for isolates MS from respiratory samples and other melting temperature curves ranged between 73.6-74.6°Cfor isolates Ms from arthirtic samples. 3- The conserved domain of the vlhA gene of M. synoviae was sequenced and analyzed for 9 isolates (4 Arthritic isolates and 5 Respiratory isolates). The results showed that, there was a complete concordance between all isolates nucleotide sequence which isolated from respiratory system except in one sample which observed point mutations in some nucleotides differ than other respiratory isolates as well as there was complete concordance between all isolates nucleotide sequence which isolated from joint. But in comparison Respiratory M.synoviae isolates with M.synoviae joint isolates , there was clear variation in nucleotide sequence which were confirm the result of HRM .In comparison with vaccine MS-H strain sequence, all isolates show clear variation from Ms-H vaccine strain .This study was demonstrated a difference between M.synoviae isolated from respiratory system and Joint and live commercial vaccine MS-H strain. Furthermore, these data indicated that changes in the vlhA gene sequence could introduce into the expressed vlhA gene amino acid codons and effective in pathogenesis rate in flocks. 4- The result of ELISA were 8 (40%) out of 20 were positive for M.synoviae. 5- For isolation of other bacteria causing arthritis the samples were cultivated on MaCconky’s agar, Eosine methylen blue agar,X.L.D agar and Mannitol salt agar media. The results revealed that other 40 samples were found positive for Escherichia coli, Staphylococcus aureus and Salmonella Entertidis isolation at a percentage of 40%, 20% and 5%, respectively. 6-Six serogroups of E.coli (O128, O125, O146,O27, O114 and O158) were detected 7-All serotypes were found positive for iss virulence gene, while ibeA virulence gene was detected in O125 and O146 serotypes only. 8-Results of PCR amplification for CAN and clfA virulence genes of S.aureus revealed that 3 out of 8 isolate 3(37.5%) were found positive for both genes .