The current study described the isolation and molecular detection of Mycoplasma gallisepticum (Mg) and Mycoplasma synoviae from tracheal swabs of diseased birds showing signs of respiratory distress in selected commercial (layer and broiler) farms and from yolk and an open air of pens of vaccinated breeder flocks in Sudan. A number of 45？Mycoplasma isolates were recovered from chickens in Khartoum, Gezira, and Equatoria states in Sudan. Of these, eight Mg and three Ms isolates were identified using growth inhibition and rapid serum agglutination (RSA) tests. The conventional PCR technique was applied to amplify 140？bp and 720？bp DNA fragments for the Mg and Ms, respectively. This research confirmed vertical and horizontal transmission of Mg from breeder farms through detection of Mg in yolk of fertile eggs and an air of pens despite previous vaccination. PCR is considered a rapid, sensitive, and cheap method and it will improve the diagnosis of Mycoplasma in chickens. 1. Introduction Avian mycoplasmosis was primarily described in turkeys in 1926 and in chickens in 1936 [1]. Mycoplasma gallisepticum (Mg) infection is usually designated as chronic respiratory disease of chickens and infectious sinusitis in turkeys. It is characterized by respiratory rales, coughing, nasal discharges, and frequently by sinusitis in turkeys by synovitis. Mycoplasma synoviae (Ms) infection is usually known as infectious synovitis, an acute-to-chronic infectious disease for chickens and turkeys involving primarily the synovial membranes of joints and tendons sheaths. However, during recent years, Ms has less frequently been associated with synovitis but more frequently associated with airsacculitis in chickens and sometimes in turkeys [2]. Both diseases are economically important, egg transmitted and hatchery disseminated diseases. They lead to tremendous economic losses in poultry production as a result of decreased hatchability and egg production, reduced quality of day-old chicks, reduced growth rate, increased costs of eradication procedures which involve site cleaning and depopulation, and increased costs of medication and vaccination [3]. The first isolation of both mycoplasmas in Sudan was reported by Khalda [4]. A recent study indicated that these organisms were prevalent, as 50.8% Mg and 57.6% Ms antibodies were recorded in chickens in the country [5]. For many years, diagnosis of avian mycoplasmosis was based on serological assays to detect antibody production and/or on isolation and identification of the organism. Serological tests include the rapid slide
References
[1]
B. R. Charlton, A. J. Bermudez, Boulianne et al., Avian Disease Manual, American association of avian pathologist, Kennett square, Pa, USA, 1996, Edited by: B. R. Carlton.
[2]
W. H. Yoder Jr., Avian Mycoplasmosis, Diagnostic Procedures in Veterinary Bacteriology and Mycologyedition, academic press, San Diego, Calif, USA, 5th edition, 1990, Edited by: Grace R. Carter, John R. Cole, Jr.
[3]
D. H. Ley, “Mycoplasma gallisepticum infection,” in Diseases of Poultry, Y. M. Saif, H. J. Barens, A. M. Fadly, J. R. Glisson, L. R. McDougald, and D. E. Swayne, Eds., pp. 722–744, Iowa State Press, Ames, Iowa, USA, 2003.
[4]
K. A. Khalda, Characterization of glucose positive Mycoplasma in with respect to Mycoplasma gallisepticum in Sudan [M.S. thesis], Faculty of Veterinary Medicine, University of Khartoum, 2002.
[5]
K. A. Khalda, S. A. Egbal, and M. M. Amal, “Enzyme linked immunosorbent assay for detection of Mycoplasma gallisepticm and Mycoplasma synoviae antibodies from chickens in the Sudan,” Sudan Journal of Veterinary Science & Animal Husbandary, vol. 51, no. 1, 2012.
[6]
S. H. Kleven, C. N. Rowland, and N. O. Olson, “Mycoplasma synoviae infection,” in Diseases of Poultry, B. W. Calnek, C. W. Beard, H. J. Barnes, W. M. Reid, and H. W. Yoder Jr., Eds., pp. 223–231, Iowa State University Press, Ames, Iowa, USA, 9th edition, 1991.
[7]
H. W. Yoder Jr., “Mycoplasma gallisepticum infection,” in Diseases of Poultry, B. W. Calnek, C. W. Beard, H. J. Barnes, W. M. Reid, and H. W. Yoder Jr., Eds., pp. 198–212, Iowa State University Press, Ames, Iowa, USA, 9th edition, 1991.
[8]
R. K. Saiki, S. Scharf, and F. Faloona, “Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia,” Science, vol. 230, no. 4732, pp. 1350–1354, 1985.
[9]
E. R. Nascimento, R. Yamamoto, K. R. Herrick, and R. C. Tait, “Polymerase chain reaction for detection of Mycoplasma gallisepticum,” Avian Diseases, vol. 35, no. 1, pp. 62–69, 1991.
[10]
L. H. Lauerman, F. J. Hoerr, A. R. Sharpton, S. M. Shah, and V. L. van Santen, “Development and application of a polymerase chain reaction assay for Mycoplasma synoviae,” Avian Diseases, vol. 37, no. 3, pp. 829–834, 1993.
[11]
M. García, N. Ikuta, S. Levisohn, and S. H. Kleven, “Evaluation and comparison of various PCR methods for detection of Mycoplasma gallisepticum infection in chickens,” Avian Diseases, vol. 49, no. 1, pp. 125–132, 2005.
[12]
H. Bagheri, A. Doosti, and A. Arshi, “Detection of Mycoplasma gallisepticum in Chaharmahal Va Bakhtiari Province poultry using PCR,” Global Veterinaria, vol. 7, no. 1, pp. 54–59, 2011.
[13]
F. Zahraa, A. Ideris, M. H. Omar, A. Bejo, and T. C. Giap, “The prevalence of Mycoplasma gallisepticum infection in chickens from Peninsular Malaysia,” Journal of Animal and Veterinary Advances, vol. 10, no. 14, pp. 1867–1874, 2011.
[14]
N. Ferguson-Noel, A. L. Victoria, and M. Farrar, “Influence of swab material on the detection of Mycoplasma gallisepticum and Mycoplasma synoviae by real-time PCR,” Avian Diseases, vol. 56, no. 2, pp. 310–314, 2011.
[15]
J. Power and F. T. Jordan, “Unilateral enlargement of the eye in chicks infected with strain of Mycoplasma gallisepticum,” Veterinary Record, vol. 99, no. 6, pp. 102–103, 1976.
[16]
F. T. Black, “Modifications of the growth inhibition test and its application to human T-mycoplasmas,” Applied Microbiology, vol. 25, no. 4, pp. 528–533, 1973.
[17]
S. H. Kleven, “Control of avian mycoplasma infections in commercial poultry,” Avian Diseases, vol. 52, no. 3, pp. 367–374, 2008.
