Isolation, Molecular Identification and in vitro Antibiotic Susceptibility Testing of Mycoplasma agalactiae From Goats in Two Provinces of Kurdistan Region-Iraq.

Rizgar R. Sulaiman

University of Sulaimania - College of Veterinary Medicine, department of clinic and internal medicine

DOI: https://doi.org/10.17656/jzs.10406

Abstract

Mycoplasma agalactiae is one of the causal agents of classical contagious agalactia

(CA). It occurs in many parts of the world and most notably in the Mediterranean

Basin. The aims of this study were isolation and molecular identification of

Mycoplasma agalactiae for the first time in Kurdistan region from goats showing

contagious agalactia and in vitro evaluation of the activities of different

antimicrobial against Mycoplasma agalctiae. During the period of January 2011 to

November 2012 a total of 126 milk samples were collected (68 from Slemani

governorate and 58 from Arbil governorate) from goats that had clinical signs of

CA. Mycoplasma agalactiae was recovered from 102(81%) milk samples out of

(126) divided as; 58 (85.3%) out of 68 milk samples in Slemani governorate were

positive for M. agalactiae while 44(75.9%) out of 58 milk samples were positive in

Arbil governorate. All isolates were confirmed by amplification of 1624 bp uvrC

gene by PCR assay. GenBank accession number of the nucleotide and amino acid

sequences of Slemani and Arbil isolates were reported in this study is (KC 594646),

(KC 594647) respectively. The isolates in current work from both governorates

were showed 99% homology to each other and the topology of the phylogenetic

tree indicated that both field isolates were clustered together and they were

belonging to sub linage that contain most of PG2 strains. Danofloxacine and

Azithromycine showed a great effectiveness against all isolates and may be

considered as standard treatment for CA in this region.

Key Words: Contagious agalactia; Mycoplasma agalactiae uvrC gene antibiotic susceptibility

References:

[1] Bergonier D, Berthelot X, Poumarat F. Contagious agalactia of small ruminants: current knowledge concerning

epidemiology, diagnosis and control. Revue scientifique et technique (International Office of Epizootics).

1997;16(3):848-73.

[2] De la Fe C, Amores J, Tardy F, Sagne E, Nouvel L-X, Citti C. Unexpected genetic diversity of Mycoplasma

agalactiae caprine isolates from an endemic geographically restricted area of Spain. BMC veterinary research.

2012;8(1):146.

[3] Gil M, Peña F, Hermoso De Mendoza J, Gomez L. Genital lesions in an outbreak of caprine contagious agalactia

caused by Mycoplasma agalactiae and Mycoplasma putrefaciens. Journal of Veterinary Medicine, Series B.

2003;50(10):484-7.

[4] Assunçao P, De la Fe C, Ramirez A, Andrada M, Poveda J. Serological study of contagious agalactia in herds of

goats in the Canary Islands. The Veterinary Record. 2004;154(22):684-7.

[5] Corrales J, Esnal A, De la Fe C, Sánchez A, Assunçao P, Poveda J, et al. Contagious agalactia in small ruminants.

Small ruminant research. 2007;68(1):154-66.

[6] Hasso S, Al-Aubaidi J, Al-Darraji A. Contagious agalactia in goats: It's severity as related to the route of infection

and pregnancy. Small Ruminant Research. 1993;10(3):263-75.

[7] Al-Aubaidi JM, Saed, O. M., Al-Shammari J. N. and Hayran, K. A. Contagious agalctia of sheep and goats,

isolation and identification of Mycoplasma agalactiae clinical and pathological confirmation of the disease. the six

international congress of the M10, PII-100, . 1986(August ):26-31.

[8] Ayling R, Baker S, Peek M, Simon A, Nicholas R. Comparison of in vitro activity of danofloxacin, florfenicol,

oxytetracycline, spectinomycin and tilmicosin against recent field isolates of Mycoplasma bovis. The Veterinary

Record. 2000;146(26):745-7.

[9] Loria G, Sammartino C, Nicholas R, Ayling R. In vitro susceptibilities of field isolates of Mycoplasma agalactiae to

oxytetracycline, tylosin, enrofloxacin, spiramycin and lincomycin–spectinomycin. Research in veterinary science.

2003;75(1):3-7.

[10] Nicholas R, Baker S. Recovery of mycoplasmas from animals. Mycoplasma protocols: Springer; 1998. p. 37-43.

[11] Subramaniam S, Bergonier D, Poumarat F, Capaul S, Schlatter Y, Nicolet J, et al. Species identification of

Mycoplasma bovis and Mycoplasma agalactiae based on the uvrC genes by PCR. Molecular and cellular probes.

1998;12(3):161-9.

[12] Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics

analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular biology and

evolution. 2011;28(10):2731-9.

[13] Hannan PC. Guidelines and recommendations for antimicrobial minimum inhibitory concentration (MIC) testing

against veterinary mycoplasma species. Veterinary research. 2000;31(4):373-95.

[14] Nicholas R, Ayling R, McAuliffe L. Contagious agalactia. Mycoplasma diseases of ruminants. 2008:98-113.

[15] Pilo P, Fleury B, Marenda M, Frey J, Vilei E M. Prevalence and distribution of the insertion element IS g1 in

Mycoplasma agalactiae. Veterinary Microbiology. 2003;92(1):37-48.

[16] Watkins G, Jones J. Mastitis and contagious agalactia. Diseases of Sheep. 2007:99-105.

[17] Damdinsuren C. Mycoplasmosis in farm animals in Mongolia: immunization of sheep and goats against

contagious agalactia. Arch Exper Vet Med. 1989;43:769-72.

[18] Kheirabadi K, Ebrahimi A. Investigation of Mycoplasma agalactiae in milk and conjunctival swab samples from

sheep flocks in west central, Iran. Pakistan journal of biological sciences: PJBS. 2007;10(8):1346-8.

[19] Kizil O, Ozdemir H. Clinical, haematological and biochemical studies in goats naturally infected with Mycoplasma

agalactiae. Bulletin of the Veterinary Research Institute in Pulawy. 2006;50(3):325-8.

[20] McAuliffe L, Gosney F, Hlusek M, de Garnica ML, Spergser J, Kargl M, et al. Multilocus sequence typing of

Mycoplasma agalactiae. Journal of medical microbiology. 2011;60(6):803-11.

[21] Sancar G, Smith FW, Lorence MC, Rupert C, Sancar A. Sequences of the Escherichia coli photolyase gene and

protein. Journal of Biological Chemistry. 1984;259(9):6033-8.

[22] Sancar A, Lindsey-Boltz LA, Ünsal-Kaçmaz K, Linn S. Molecular mechanisms of mammalian DNA repair and the

DNA damage checkpoints. Annual review of biochemistry. 2004;73(1):39-85.

[23] Garnica M, Rosales R, Gonzalo C, Santos J, Nicholas R. Isolation, molecular characterization and antimicrobial

susceptibilities of isolates of Mycoplasma agalactiae from bulk tank milk in an endemic area of Spain. Journal of

applied microbiology. 2013;114(6):1575-81.

[24] Manso-Silván L, Dupuy V, Lysnyansky I, Ozdemir U, Thiaucourt F. Phylogeny and molecular typing of

Mycoplasma agalactiae and Mycoplasma bovis by multilocus sequencing. Veterinary microbiology. 2012;161(1):104-12.

[25] Sarris K. Contagious agalactia. Frey, J and Sarris, K: Mycoplasmas of ruminants: pathogenicity, diagnostics,

epidemiology and molecular genetics, COST. 1996;826:12-5.

[26] Al-Momani W, Halablab MA, Abo-Shehada MN, Miles K, McAuliffe L, Nicholas RA. Isolation and molecular

identification of small ruminant mycoplasmas in Jordan. Small Ruminant Research. 2006;65(1):106-12.

[27] Azevedo EOd, Alcântara MDBd, Nascimento ERd, Tabosa IM, Barreto ML, Almeida JFd, et al. Contagious

agalactia by Mycoplasma agalactiae in small ruminants in Brazil: first report. Brazilian Journal of Microbiology.

2006;37(4):576-81.

[28] Mohammadpour SH, Pourbakhsh SA, Kheirkhah B. Isolation and identification of Mycoplasma agalactiae by

polymerase chain reaction (PCR) in suspected sheep samples in Kerman Province, Iran. African Journal of

Microbiology Research. 2013;7(10):885-9.

[29] Paterna A, Sánchez A, Amores J, Gómez-Martín Á, Corrales JC, Contreras A, et al. Survival of< i> Mycoplasma

agalactiae and Mycoplasma mycoides subspecies capri in heat treated goat colostrum. The Veterinary Journal.

2013;196(2):263-5.

[30] Ayling R, McAuliffe L. Mycoplasma diseases of ruminants: CABI; 2008.

[31] Hirsh DC. Mycoplasmas. Pages 466–468 in Antimicrobial Therapy in Veterinary Medicine. J. F. Prescott, J. D.

Baggot, and R. D. Walker, ed. Iowa State University Press, Ames. 2000.

[32] Furneri PM, Rappazzo G, Musumarra MP, Di Pietro P, Catania LS, Roccasalva LS. Two New Point Mutations at

A2062 Associated with Resistance to 16-Membered Macrolide Antibiotics in Mutant Strains of Mycoplasma hominis.

Antimicrobial agents and chemotherapy. 2001;45(10):2958-60.

[33] Lysnyansky I, Mikula I, Gerchman I, Levisohn S. Rapid detection of a point mutation in the parC gene associated

with decreased susceptibility to fluoroquinolones in bovis. Antimicrobial agents and chemotherapy. 2009;53(11):4911-4.

[34] Antunes N, Tavío M, Assunção P, Rosales R, Poveda C, De la Fe C, et al. In vitro susceptibilities of field isolates

of Mycoplasma agalactiae. The Veterinary Journal. 2008;177(3):436-8.

View Full Article