Huban Göçmen, Esra Buyukcangaz, Burcu Ustuner, Sevil Erdenlig, Selim Alcay, M. Berk Toker, Engin Kennerman, Mihriban Ulgen

Koyun ve Saanen keçilere ait semenlerden sistematik deteksiyon ile kronik Bruselozisin tanısı

Çalışma, 2012-2014 yılları arasında Türkiye'nin kuzeybatısında yer alan Bursa ilinde brucellosis geçmişli keçi (n: 206) ve koyunlara (n: 38) ait bir sürüden (n: 244) örneklemeler suretiyle yapıldı. Saanen keçilerinden (n: 35) ve koçlardan (n: 8) alınan semen örnekleri Brucella spp. ve diğer zoonotik bakteriyel ajanların tespiti amacıyla çalışıldı. Örnekler, rutin tanısal prosedürler ve PCR ile test edildi. Erkek hayvanlardan alınan serum örnekleri ayrıca C-ELISA ve I-ELISA ile Bruselloz yönünden test edildi. Kültür sonuçları; Semen örneklerinden Trueperella pyogenes (n: 2), Pasteurella pneumotopica (n: 5), Esherichia coli (n: 3), Aeromonas salmonicida sub. Salmonicida (n:1), Brevundimonas vesicularis (n: 2) ve Mycoplasma bovigenitalium (n: 1) ve Mycoplasma arginini (n: 1) izole edildi. Klinik muayenede ise koçlarda epididimitis veya epididimoorşit kaynaklı semptom olmamasına karşın serolojik olarak bruselloz pozitif olan iki adet koç orşitis idi. Ayrıca bir seronegatif koç epididimitis semptomu gösterdi. Semen örneklerinin muayenesinde serolojik olarak pozitif ve negatif hayvanlar arasında akrozomun hacmi, konsantrasyonu, kütle aktivitesi, motilitesi ve defektivite oranı açısından istatistiksel olarak anlamlı bir fark bulunmadı. Serum örneklerinden 20 adedi anti-Brucella antikoru için negatif olmasına rağmen, 23 adedi bruselloz için serolojik olarak pozitifti. Selektif besiyerinde yapılan katı ve sıvı kültürler ile semen örneklerinden yapılan tür spesifik PCR testi sonucunda örneklerden Brucella saptanmadı. Bu durum, numunelerin; hayvanların genellikle organizmaları saçmadığı kronik enfekte hayvanlardan toplanmış olabileceğini göstermektedir. Periyodik örneklemenin brusellozun kesin tanısında ve insidensinin saptanmasında yardımcı olabileceği düşünülmektedi

A Systematic Detection for Brucellosis at Chronic Stage of Infection in Semen of Sheep and Saanen Goats

The study was conducted in a herd (n: 244) in which goats (n: 206) and sheep (n:38) had a history of brucellosisin Bursa which is located in Northwestern of Turkey between the years 2012-2014. For the detection of Brucellaspp. and the other zoonotic bacterial agents, semen samples were taken from Saanen goats (n: 35) and rams (n:8). Samples were tested by routine diagnostic procedures and PCR. The serum samples of male animals werealso tested for Brucellosis by C-ELISA and I-ELISA. The culture results represented Trueperella pyogenes (n:2),Pasteurella pneumotopica (n: 5), Esherichia coli (n: 3), Aeromonas salmonicida subs. Salmonicida (1),Brevundimonas vesicularis (n: 2) and Mycoplasma bovigenitalium (n: 1) and Mycoplasma arginini (n: 1) fromsemen samples. Rams had no symptoms due to epididymitis or epididymoorchitis in clinical examination, buttwo bucks showed orchitis and they were serologically positive for brucellosis. Also, one seronegative buckshowed epididymitis in a flock. There were no statistically significant differences between the serologicallypositive and negative animals in an examination of semen samples in terms of their volume, concentration, massactivity, motility and defectivity rate for acrosome. Although 20 of the serum samples were negative for antiBrucella antibody, 23 of them were serologically positive for brucellosis. As a result of this study, Brucellaewere not detected by bacteriologically and molecularly while there were some positive serum samples forbrucellosis. This could be attributed that these samples might have been collected from chronically infectedanimals in which animals generally do not shed the organisms. Therefore, it was thought that sampling withregular intervals might help for the definitive incidence of brucellosis.

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Alton GG, Jones LM, Angus RD, Verger JM. 1988. Bacteriological methods. In: Techniques for the Brucellosis Laboratory. INRA. pp 13-61. Conference Animal Diseases 2009, Royal Holloway, University of London UK, 2-4 September.
Alton Godfrey G. 1990. Edited by Nielsen Klauss, Duncan J. Robert, Animal Brucellosis, Chapter 16 Brucella melitensis, 1887 to 1997, Chapter 17 Brucella melitensis, pp:379--411), CRC Press, Boca Raton, USA, 1990.
Andreani E. 1967.Decorso dell’infezione brucellare in montoni sotoposti a contagio naturale. Ann Fac Med Vet Pisa, 19, 331.
Adel S, Amina AS, Mahmoud ER, Hamdyb, Adel KI. 2001. Detection of Brucella melitensisin semen using the polymerase chain reaction assay. Veterinary Microbiology. 83(1): 37-44 https://doi.org/10.1016/S0378-1135(01)00401-1
Baily GG, Krahn JB, Drasar BS, Stoker NG. 1992. Detection of Brucella melitensis and Brucella abortus by DNA amplification. Journal of Tropical Medice and Hygien 95:271–275.
Ball L, Young S, Carroll EJ. 1968, Seminal vesiculitis syndrome: lesion in genital organs of young bulls. Am J Vet Res 29:1173–1184.
Bauerfeind R, von Graevenitz A, Schiefer AKHG, Slenczka TSW, Zahner H. 2016. Zoonoses Infectious Diseases Transmissible Between Animals and Humans, Pp:191-195, 2
Bacterial Zoonoses, 2.5 Brucellosis. ISBN 978-1-55581-925- 5 e-ISBN 978-1-55581-926-2 doi:10.1128/9781555819262, ASM Press, 1752 N St., N.W., Washington, DC 20036-2904, USA
Blasco JM, 1990. Chapter 15 Brucella ovis, Edited by Nielsen Klauss, Duncan J. Robert, Animal Brucellosis, pp:351-378
Blasco JM. 1992. Diagnosis of Brucella melitensis infection in small ruminants. In: Plommet, M. (Ed.), 247 Prevention of Brucellosis in the Mediterranean Countries. Pudoc Scientific Publishers, Wageningen, The 248 Netherlands, pp. 272-278.
Bricker BJ, Hallıng SM. Journal of Clinical Microbiology, Nov. 1994, American Society for Microbiology. Differentiation of Brucella abortus bv. 1, 2, and 4, Brucella melitensis, Brucella ovis, and Brucella suis bv. 1 by PCR. 32(11): 2660-2666.
Cameron RD, Lauerman LH. 1976.Characteristics of semen changes during Brucella ovis infection in Rams., Veterinary Record. 99(12): 231-233, DOI: 10.1136/vr.99.12.231
Carvalho Júniora CA, Moustacasa VS, Xaviera MN, Costaa EA, Costaa LF, Silvaa TMA, Paixãob TA, Borgesa AM, Gouveiac AMG, Santos RL. 2012. Andrological, pathologic, morphometric, and ultrasonographic findings in rams experimentally infected with Brucella ovis. Small Ruminant Research Volume 102, Issues 2–3, February 2012, Pages 213- 222 https://doi.org/10.1016/j.smallrumres.2011.08.004.
Corbel MJ. 1985.Recent advances in the study of Brucella antigens and their cross reactions. The Veterinary bulletin 55: 927-94
Catania S, Tavella A, Gobbo F, Ashley R, Nicholas J. 2014. The isolation of Mycoplasm bovigenitalium from infertile dairy cattle.Veterinary Record Case Reports, 2(1): 1–2
Carrera-Chávez JM, Quezada-Casasola A, Pérez-Eguia E, ItzáOrtíz MF, Gutiérrez-Hernández JL. 2016. Sperm quality in naturally infected rams with Brucella ovis, Small Ruminant Research. 144: 220–224. https://doi.org/10.1016 /j.smallrumres.2016.09.021
Crawford RP, Huber JD, Adams BS, Chapter 7 Epidemiology and Surveillence, Edited by Edited by Nielsen Klauss, Duncan J. Robert, Animal Brucellosis, pp:131- 152, CRC Press, Boca Raton, USA, 1990.
Corrales JC, Esnal A, Sánchez A, Assunçao P, Poveda JB, Contreras A. 2007. Contagious agalactia in small ruminants. Small Ruminant Research, 68 (1–2): 154–166. https://doi.org/10.1016/j.smallrumres.2006.09.010
Dartnell LR, Hunter SJ, Lovell KV, Coates AJ, Ward JM. 2010. “Low-Temperature Ionizing Radiation Resistance of Deinococcus radiodurans and Antarctic Dry Valley Bacteria”. Astrobiology, 10(7): 717-32. Bibcode: 2010AsBio10.717D.doi:10.1089/ast.2009.0439.PMID20950 171.
Dénes B, Glávits R. 1994. Bacteriologically confirmed cases of ovine epididymo-orchitis caused by Brucella ovis in SubCarpathia. Acta Vet Hungaria.;42(1): 25-33.
Eaglesome MD, Garcia MM, Stewart RB. 1992. Microbial agents associated with bovine genital tract infections and semen. Part II. Haemophilus somnus, Mycoplasma spp and Ureaplasma spp, Chlamydia; pathogens and semen contaminants; treatments of bull semen with antimicrobial agents. The Veterinary Bulletin, 62: 887-910
Enright FM. 1990. The pathogenesis and pathobiology of infection in domestic animals. Animal brucellosis.In: Nielsen & Duncan, Eds. CRC Press, Boca Ratón: Florida, USA, 301- 320.
Fekete A, Bantle JA, Halling SM, Sanborn MR. 1990. Preliminary development of a diagnostic test for Brucella using polymerase chain reaction. Journal of Applied Bacteriology, 69(2): 216-27.
Ficapal A, Jordana J, Blasc JM, Moriyón I. 1998. Diagnosis and epidemiology of Brucella ovis infections in rams. Small Rumin. Res. 29: 13–19
Fish NA, Rosendal S, Miller RB. 1985. The distribution of Mycoplasmas and Ureaplasmas in the genital tract of normal artificial insemination bulls. The Canadian Veterinary Journal, v.26, pp.13-15
Gouletsou G, Fthenakis GC. 2015. Microbial diseases of the genital system of rams or bucks P. Vet Microbiol, Dec 14;181(1-2):130-5. doi: 10.1016/j.vetmic.2015.07.016. Epub 2015 Jul 15.
Hamdy MER, Amın AS. 2002. Detection of Brucella Species in the Milk of Infected Cattle, Sheep, Goats and Camels by PCR The Veterinary Journal, 163: 299-305 doi:10.1053 /tvjl.2001.0681
Jubb KVF, Kennedy PC, Palmer N. 1993. Pathology of domestic animals. 4th ed. Academic Press: San Diego, California, USA, 3: 653.
Kaushik P, Singh DK, Tiwaril AK, Kataria RS. 2006. Rapid Detection of Brucella Species in Cattle Semen by PCR. J. Appl. Anim. Res. 30: 25-28.
Khamesipour F, Doosti A, Taheri H. 2013. Molecular Detection of Brucella Spp. İn The Semen, Testis and Blood Samples of Cattle And Sheep, Journal of Pure and Applied Microbiology, 7(Spl. Edn.): 495-
Lambert M. 1987. Contagious agalactia of sheep and goats. Rev Sci Tech Off Int Epiz,. 6(3): 699–711.
Nicholas RAJ, Wessels M, Orme PK, Wood E, Sachse K. 1999. Isolation of Mycoplasma ovine/caprine serogroup 11 from infertile sheep in Britain. Veterinary Record 145: 434-435
Leon CF. 1994. Brucelosis ovina y caprina, Ed. Office International dês Epizooties – OIE. Paris: Franci: 451. Inhart RD, Parker WG. 1988, Seminal vesiculitis in bulls. Compend Cont Ed Pract Vet 10: 1429–1448.
Mejid J, Mathias LA, Robles CA. 2010. Clinical manifestations of brucelosis endomestic animals and humans. Open Vet. Sci. 4: 119–126.
Morgan WJB, MacKinnon DJ, 1979. Brucellosis, In laining, JA,Fertility and infertility in domestic animals, 3rd edition, ELBS, Bailliere Tindall, Chapter 9: 171-198.
Moro BEMP, Weiss RDN, Friedrich RSC, Vargas AC, Weiss LHN, Nunes MP. 1999. Aeromonas hydrophila isolated from cases ofbovine seminal vesiculitis in South Journal of Veterinary Diagnostic Investigation 11: 189–191.
Nicholas RA, Khan LA, Houshaymi B, Miles RJ, Ayling RD, Hotzel H, Sachse K. 2002. Close genetic and phenotypic relatedness between Mycoplasma ovine/caprine serogroup 11 and Mycoplasma bovigenitalium. Systematic and Applied Microbiology, 25(3): 396–402.
OIE. 2009. Office International dês Epizooties, Manual of Standards for Diagnostic tests and Vaccines. Third edition. Office International of Epizooties, Paris, France. Caprine and ovine brucellosis. Chapter (2.7.2), Bovine Brucellosis chapter (2.4.3).
Poveda J. 1998. Biochemical Characteristics in Mycoplasma Identification. In: Miles, R, Nicholas, R. (Org.). Mycoplasma Protocols. New Jersey, Totowa: Humana Press Inc, V. 104: 37–50: 69–78.
Prats-van der Ham M, Tatay-Dualde J, Fe C, Paterna A, Sánchez A, Corrales JC, Contreras A. Gómez-Martín A. 2017. Detecting asymptomatic rams infected with Mycoplasma agalactiae in ovine artificial insemination centers, Theriogenology 89: 324–328.
Ridler AL, West DM. 2011. Control of Brucella ovis Infection in Sheep, Vet Clin Food Anim 27 61–66 doi:10.1016 /j.cvfa.2010.10.013
Quinn PJ, Markey BK, Carter ME, Donnely WJ, Leonard FC, 2003. Chapter 78, Interactions of microbial pathogens with the male and female reproductive systems, pp:450-456, in Veterinary Microbiology and Microbial Diseases. Blackwell publishing, ISBN:0-632-05525.
Ustuner B, Alcay S, Toker MB, Nur Z, Gokce E, Sonat FA, Gul Z, Duman M, Ceniz C, Uslu A, Sagirkaya H, Soylu MK. 2016. Effect of rainbow trout (Oncorhynchus mykiss) seminal plasma on the post-thaw quality of ram semen cryopreserved in a soybean lecithin-based or egg yolk-based extender. Anim Reprod Sci 164: 97-104.
Van Metre DC, Rao S, Kimberling CV, Morley PS. 2012. Factors associated with failure in breeding soundness examination of Western USA rams. Prev. Vet. Med. 105: 118–126.
Von Beroldingen CH, Blake ET, Higuchi R, Sensabaugh GF, Erlich HA. 1990. Application of PCR to 284 the analysis of biological evidence. In: Erlich, H.A. (Ed.), 1, PCR Technology: Principles and Applications 285 for DNA Amplification. Stockton Press, New York, pp. 209-223.
Xavier MN, Azebedo CE, Alves PT, Lima SR. 2009. The genus Brucella and clinical manifestations of brucelosis. Ciência Rural 39: 2252–2260.