Manuscript Title:

PREVALENCE AND PHYLOGENETIC ANALYSIS OF MYCOPLASMA GALLISEPTICUM ISOLATES IN BROILERS AND LAYERS

Author:

MUHAMMAD ASIF, SAJJAD UR RAHMAN, MUHAMMAD IMRAN ARSHAD, FARRAH DEEBA

DOI Number:

DOI:10.17605/OSF.IO/JRDVA

Published : 2023-03-10

About the author(s)

1. MUHAMMAD ASIF - Institute of Microbiology, Faculty of Veterinary sciences, University of Agriculture Faisalabad, Pakistan.
2. SAJJAD UR RAHMAN - Institute of Microbiology, Faculty of Veterinary sciences, University of Agriculture Faisalabad, Pakistan.
3. MUHAMMAD IMRAN ARSHAD - Institute of Microbiology, Faculty of Veterinary sciences, University of Agriculture Faisalabad, Pakistan.
4. FARRAH DEEBA - Department of Clinical Medicine and Surgery, Faculty of Veterinary sciences, University of Agriculture Faisalabad, Pakistan.

Full Text : PDF

Abstract

Mycoplasma gallisepticum (M. gallisepticum) is a bacterial pathogen and causes economic losses in poultry. It is responsible for acute respiratory disease which stay without clinical signs but the infected birds become prone to secondary infections. The study was aimed for molecular detection and phylogenetic analysis of M. gallisepticum collected from commercial poultry birds (broilers and layers). Total 400 swab samples were collected and analyzed through serum plate agglutination test for seroprevalence followed by culturing on Frey’s medium. Culture positive samples were furthered processed by PCR and sequenced for phylogenetic study. From seroprevalence results, it was found that the positive samples showed agglutination in serum plate agglutination test. The seroprevalence results of M. gallisepticum indicated that layers are more positive (31.9%, n=210) than broilers (24.7%, n=190). In culture identification, positive isolates depicted typical fried egg-shaped colonies of M. gallisepticum on modified Frey’s agar medium. Based on colony morphology and biochemical characterization, 15.24% of layer samples were positive while broiler samples showed low positivity rate (9.47%). Through PCR, culture positive isolates indicated the product of 185bp, confirming the specie of M. gallisepticum. In comparative prevalence of PCR positive samples, it was found that layer samples were more positive (20.48%) than broiler samples (13.16%). Samples collected from various organs of broilers and layers were processed separately through culturing and PCR. It was found that tracheal samples showed higher prevalence than other organ samples of broilers. While the same trend was observed in layer samples collected from various organs. In phylogenetic analysis, sequences with more than eighty percent similarity were analyzed by MEGA X for evolutionary history. A total of 871 positions were observed in the final data which was submitted to GenBank. The phylogenetic analysis of M. gallisepticum from the present study (MZ373234.1, MZ373235.1, and MZ424735.1) showed genetic sequence mimicry to the other isolates. In conclusion, layers could be the reservoir for M. gallisepticum infection and new genetic methods are needed for confirmation and differentiation of new emerging strains.


Keywords

Prevalence, Phylogenetic Analysis, Genetic Methods.