Sequencing Based Phylogenetic Analysis of Local Mycoplasma Gallisepticum of Broiler Chickens in Al-Dewaniyah Province / Iraq

Abstract

Mycoplasma gallisepticum (MG) is regarded as finest bacteria can replicate independently and they one of the illustrious pathogens for chickens cause body retardation and losing weight thereby it considered as one of the costliest worldwide significant pathogens for poultry. Mycoplasma detection using traditional culture method is not adequate procedure because it has several strains also the difficulty of cultivation technique and the obstructs that faced, current study was considered culture method and conventional Pcr assay for direct diagnosis of MG using 16S rRNA and mgc1(GapA) gene.

Out of 20 commercial broiler farms at Al-dewaniyah province, Iraq, 150 tissue specimens were assembled wherever manifestation of respiratory infections was demonstrative.   

Whereas 16 out of 150 samples (10.66 %) were recognized as positive via conventional cultivation based on morphology of growing colonies, Diene’s staining and some biochemical, however confirmed by amplification of 16S rRNA gene of polymerase chain reaction technique of suspected colonies, while cloning of direct tissue samples by 16S rRNA amplification exhibit 36/150 (24%) corresponding to the genus of Mycoplasma, the 16S rRNA amplification product was 1500 base pair.

            The inference of the findings of current study, is that, the PCR revealed greater sensitivity and viable reliability, quality and precision than bacterial culture techniques and therefore might be very suggestive for the supervision of flock’s health imperviousness against MG and to enable application of effective preventative and control measures. all 16 cultured isolates that affirmative as Mycoplasma spp by 16S rRNA amplification were undergo by PCR analysis of GapA gene in addition to sequence analysis of 16S rRNA, the outcomes of 16 isolated Mycoplasma spp. colonies showed that 8 isolates were belongs to Mycoplasma gallisepticum out of 150 total tested samples with percent of (5.33%), this result provided by virulence GapA (mgc1) gene analysis data, however positive tissue samples by cloning assay was 22/150 (14.66%) were belongs to M. gallisepticum. Based on results of 16S rRNA sequencing, we detect other Mycoplasma spp. and other unrelated bacteria not showed in the current article. The results of Mycoplasma isolates approval through amplification of the GapA gene showed a bands of 332 bp for M. gallisepticum, the16S rRNA gene was directed to Soul University (Korea) for sequence analysis. The available data are presented in Gene bank database of 16S rRNA for documentation.