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Identification and antimicrobial susceptibility of Corynebacterium pseudotuberculosis isolated from sheep

Year 2015, Volume: 31 Issue: 2, 116 - 121, 01.06.2015

Abstract

Aim: The aim of this study was to identification of Corynebacterium pseudotuberculosis (C. pseudotuberculosis) isolated from lymph node of sheep with caseous lypmhadenitis (CLA) by biochemical characteristics and confirmation by Polymerase Chain Reaction (PCR). Also, phospholipase D enzyme activities and antimicrobial susceptibility of isolates were determined. Materials and Methods: A total of 1176 sheep were examined the presence of CLA, and 1176 of lymph node were collected for bacteria isolation. The samples were inoculated onto Blood Agar Base supplemented with 5% defibrinated sheep blood at 37ºC for 48 hours in microaerophilic atmosphere. Morphological and biochemical characteristic of C. pseudotuberculosis suspicious isolates were determined. PCR analysis was performed to confirmation of biochemical identification. Also, phospholipase D enzyme activities with Rhodococcus equi (R. equi) and Staphylococcus aureus (S. aureus) and antimicrobial susceptibility of isolates for 16 antimicrobial were determined by disc diffusion methods.Results: Totally of 72 C. pseudotuberculosis were identified according to biochemical characteristics. All isolates showed PLD enzyme activities antagonists with S. aureus and synergistically with R. equi. All isolates were confirmed by PCR as C. pseudotuberculosis. Isolates were determined to be sensitive to different antibiotics at different rates. The majority of the isolates were susceptible to florfenicol (98%), while ampicillin and lincomycin resistance was determined as 62.5%.Conclusions: Although there are many data related to the failure of the in-vivo treatment of CLA infection by antibiotics, C. pseudotuberculosis was determined as in-vitro sensitive to many antibiotics used in this study. PCR method and primers used in this study was determined very useful for confirmation of C. pseudotuberculosis identified by biochemical characteristic.

References

  • Al-Gaabary MH, Osman SA, Ahmed MS, Oreiby AF, 2010. Abattoir survey on caseous lymphadenitis in sheep and goats in Tanta, Egypt. Small Rum. Res, 94, 117-124.
  • Al-Gaabary MH, Osman SA, Oreiby AF, 2009. Caseous lymphadenitis in sheep and goats: Clinical, epidemiological and preventive studies. Small Rum Res, 87, 116-121.
  • Arsenault J, Girard C, Dubreuil P, Daignault D, Galarneau J, Boisclair J, Simard C, Belanger D, 2003. Prevalence of and carcass condemnation from maedi-visna, paratuberculosis and caseous lymphadenitis in culled sheep from Quebec, Canada. Prev Vet Med, 59, 67-81
  • Baird GJ, 2006. Treatment of ovine caseous lymphadenitis. Vet Rec, 159, 500.
  • Baird GJ, Fontaine MC, 2007. Corynebacterium pseudotuberculosis and its role in ovine caseous lymphadenitis. J Comp Path, 137, 179-210.
  • Bauer AW, Kirby WMM, Sherris JC, Turck M, 1966. Antibiotic susceptibilty testing by a standardized single disk method. Am J Clin Path, 45, 493.
  • Cetinkaya B, Karahan M, Atil E, Kalin R, De Baere T, Vaneechoutte M, 2002. Identification of Corynebacterium pseudotuberculosis isolated from sheep and goats by PCR. Vet Microbiol, 88, 75-83.
  • Costa LRR, Spier SJ, Hirsh D, 1998. Comparative molecular characterization of Corynebacterium pseudotuberculosis of different origin. Vet Microbiol, 62, 135-143.
  • Dercksen DP, Brinkhof JMA, Toos Dekker-Nooren T, van Maanen K, Bode C F, Baird G, Kamp EM, 2000. A comparison of four serological tests for the diagnosis of caseous lymphadenitis in sheep and goats. Vet Microbiol, 75, 167-175.
  • Doherr MG, Carpenter TE, Wilson WD, Gardner IA, 1998. Application and evaluation of a mailed questionnaire for an epidemiologic study of Corynebacterium pseudotuberculosis infection in horses. Prev Vet Med, 35, 241-253.
  • Erganiş O, Kaya O, Ateş M, İstanbulluoglu E, 1990. Konya EBK kombinasında kesilen koyunlardaki apseli lenf yumruları üzerine mikrobiyolojik ve serolojik incelemeler. Veterinarium, 1, 8-11.
  • Esterabadi AH, Entessar F, Hedayati H, Narimani AA, Sadr IM, 1975. Isolation of Corynebacterium pseudotuberculosis from camel in Iran. Archives de L’Institut Razi, 27, 61-66. Fontaine MC, Baird GJ, 2008. Caseous lymphadenitis. Small Rum Res, 76, 42-48.
  • Funke G, Bernard, KA. 2003. Coryneform gram-positive rods, in: Manual of Clinical Microbiology, Eds: Murray PR, Baron EJ, Jorgensen JH, Pfaller MA, Yolken RH, ASM Press, Washington, USA, pp: 472-501.
  • Funke G, von Graevenitz A, Clarridge JE, Bernard KA, 1997. Clinical microbiology of coryneform bacteria. Clin Microbiol Rev, 10, 125-159.
  • Guimaraes AS, Seyffert N, Bastos BL, et al., 2009. Caseous lymphadenitis in sheep flocks of the state of Minas Gerais, Brazil: Prevalence and management surveys. Small Rum Res, 87, 86-91.
  • Hawari AD, 2008. Corynebacterium pseudotuberculosis infection (Caseous Lymphadenitis) in camels (Camelusdromedarius) in Jordan. Am J Anim Vet Sci, 3, 68-72.
  • Hodgson ALM, Tachedjian M, Corner LA, Radford AJ, 1994. Protection of sheep against caseous lymphadenitis by use of a single oral dose of live recombinant Corynebacterium pseudotuberculosis. Infect Immun, 62, 5275-5280.
  • Ilhan Z, 2013. Detection of Corynebacterium pseudotuberculosis from sheep lymph nodes by PCR. Revue Med Vet, 164, 2, 60-66.
  • Izgür M, Akan M, İlhan Z, 1999. Microorganisms isolated from cases of caseous lymphadenitis. Vet J Ankara Univ, 46, 43-50.
  • Judson R, Songer JG, 1991. Corynebacterium pseudotuberculosis: In-vitro susceptibility to 39 antimicrobial agents. Vet Microbiol, 27, 145-150.
  • Literak I, Horvathova A, Jahnova M, Rychlik L, Skalka B, 1999. Phenotype and genotype characteristics of the Slovak and Czech Corynebacterium pseudotuberculosis strains isolated from sheep and goats. Small Rum Res, 32, 107-111.
  • Lopez JF, Wong FM, Quesada J, 1966. Corynebacterium pseudotuberculosis: First case of human infection. Am J Clin Pathol, 46, 562-567.
  • Muckle CA, Gyles CL, 1982. Characterization of Strains of Corynebacterium pseudotuberculosis. Can J Comp Med, 46, 206-208.
  • NCCLS, 2003. National committee for clinical laboratory standards: Performance standards for antimicrobial susceptibility testing, 8th edition, NCCLS document M2-A8 volume 23, no 1, USA.
  • O’Reilly KM, Green LE, Malone FE, Medley GF, 2008. Parameter estimation and simulations of a mathematical model of Corynebacterium pseudotuberculosis transmission in sheep. Prev Vet Med, 83, 242-259.
  • Pacheco LGC, Pena RR, Castro TLP, et al., 2007. Multiplex PCR assay for identification of Corynebacterium pseudotuberculosis from pure cultures and for rapid detection of this pathogen in clinical samples. J Med Microbiol, 56, 480- 486.
  • Paton MW, Mercy AR, Sutherland SS, Ellis TM, 1988. The influence of shearing and age in the incidence of caseous lymphadenitis in Australian sheep flocks. Acta Vet Scand, 84, 101-103.
  • Paton MW, Walker SB, Rose IR, Watt GF, 2003. Prevalence of caseous lymphadenitis and usage of caseous lymphadenitis vaccines in sheep flocks. Aust Vet J, 81, 91-95.
  • Peel MM, Palmer GG, Stacpoole AM, Kerr TG, 1997. Human lymphadenitis due to Corynebacterium pseudotuberculosis: Report of ten cases from Australia and review. Clin Infect Dis, 24, 185-191.
  • Pepin M, Paton M and Hodgson AL, 1994. Pathogenesis and epidemiology of Corynebacterium pseudotuberculosis infection in sheep. Curr Topics Vet Res, 1, 63-82.
  • Prescott JF, Menzies PI, Hwang YT, 2002. An interferon-gamma assay for diagnosis of Corynebacterium pseudotuberculosis infection in adult sheep from a research flock. Vet Microbiol, 88, 287-297.
  • Quinn PJ, Carter ME, Markey BK, Carter GR, 1984. Clinical veterinary microbiology, Mosby International Ltd., Spain, pp: 118-345.
  • Seyffert N, Guimaraes AS, Pacheco LGC, et al., 2010. High seroprevalence of caseous lymphadenitis in Brazilian goat herds revealed by Corynebacterium pseudotuberculosis secreted proteins-based ELISA. Res Vet Sci, 88, 50-55.
  • Ter Laak EA, Bosch J, Bill GC, Schreuder BEC, 1992. Doubleantibody sandwich enzyme-linked immunosorbent assay and immunoblot analysis used for control of caseouslphadenitis in goats and sheep. Am J Vet Res, 53, 1125-1132.
  • Yeruham I, Elad D, Perl S, Ram A, 2003. Necrotic ulcerative dermatitis on the heels of heifers in a dairy herd infected with Corynebacterium pseudotuberculosis. Vet Rec, 152, 598-600.
  • Yeruham I, Elad D, van Ham M, Shpigel NY, Perl S, 1997. Corynebacterium pseudotuberculosis infection in Israeli cattle: Clinical and epidemiological studies. Vet Rec, 140, 423- 427.

Koyunlardan izole edilen Corynebacterium pseudotuberculosis suşlarının identifikasyonu ve antibiyotiklere duyarlılıkları

Year 2015, Volume: 31 Issue: 2, 116 - 121, 01.06.2015

Abstract

Amaç: Bu çalışmada, Kazeöz Lenfadenetis (KLA)’li koyunların lenf düğümlerinden izole edilen Corynebacterium pseudotuberculosis (C. pseudotuberculosis) izolatlarının biyokimyasal identifikasyonu, Polimeraz Zincir Reaksiyonu (PZR) ile doğrulanması, fosfolipaz D (PLD) enzim aktivitesinin belirlenmesi ve antibiyotiklere duyarlılıklarının tespit edilmesi amaçlandı. Gereç ve Yöntem: Toplam 1176 adet koyun KLA yönünden muayene edildi ve 1176 lenf düğümü bakteri izolasyonu için alındı. Örnekler, kanlı agara ekildi ve 37°C’de 48 saat mikroaerofilik olarak inkübe edildi. Şüpheli izolatların, morfolojik, biyokimyasal özellikleri PZR analizleri, Rhodococcus equi (R. equi) ve Staphylococcus aureus (S. aureus) ile PLD enzim aktiviteleri belirlendi. Ayrıca, C. pseudotuberculosis izolatlarının 16 antibiyotiğe duyarlılıkları disk diffüzyon metodu ile belirlendi. Bulgular: Toplam 72 şüpheli izolat biyokimyasal özelliklerine göre C. pseudotuberculosis olarak identifiye edildi. Tüm izolatlar S. aureus ile antagonist ve R. equi ile sinerjik olarak PLD enzim aktivitesi gösterdi. İzolatların tümü PZR ile C. pseudotuberculosis olarak doğrulandı. C. pseudotuberculosis izolatlarının farklı antibiyotiklere değişik oranlarda duyarlı oldukları, izolatların büyük çoğunluğunun (%98) florfenikole duyarlı, %62.5’inin ampisilin ve linkomisine dirençli olduğu belirlendi. Öneri: Enfeksiyonun antibiyotiklerle in-vivo tedavide başarısızlığı ile ilgili çok sayıda veri bulunmasına rağmen etkenin bu çalışmada kullanılan pek çok antibiyotiğe in-vitro duyarlı olduğu, PZR'ın ve bu çalışmada kullanılan primerlerin, biyokimyasal özelliklerine göre identifiye edilen izolatların doğrulanmasında faydalı olduğu ifade edilebilir.

References

  • Al-Gaabary MH, Osman SA, Ahmed MS, Oreiby AF, 2010. Abattoir survey on caseous lymphadenitis in sheep and goats in Tanta, Egypt. Small Rum. Res, 94, 117-124.
  • Al-Gaabary MH, Osman SA, Oreiby AF, 2009. Caseous lymphadenitis in sheep and goats: Clinical, epidemiological and preventive studies. Small Rum Res, 87, 116-121.
  • Arsenault J, Girard C, Dubreuil P, Daignault D, Galarneau J, Boisclair J, Simard C, Belanger D, 2003. Prevalence of and carcass condemnation from maedi-visna, paratuberculosis and caseous lymphadenitis in culled sheep from Quebec, Canada. Prev Vet Med, 59, 67-81
  • Baird GJ, 2006. Treatment of ovine caseous lymphadenitis. Vet Rec, 159, 500.
  • Baird GJ, Fontaine MC, 2007. Corynebacterium pseudotuberculosis and its role in ovine caseous lymphadenitis. J Comp Path, 137, 179-210.
  • Bauer AW, Kirby WMM, Sherris JC, Turck M, 1966. Antibiotic susceptibilty testing by a standardized single disk method. Am J Clin Path, 45, 493.
  • Cetinkaya B, Karahan M, Atil E, Kalin R, De Baere T, Vaneechoutte M, 2002. Identification of Corynebacterium pseudotuberculosis isolated from sheep and goats by PCR. Vet Microbiol, 88, 75-83.
  • Costa LRR, Spier SJ, Hirsh D, 1998. Comparative molecular characterization of Corynebacterium pseudotuberculosis of different origin. Vet Microbiol, 62, 135-143.
  • Dercksen DP, Brinkhof JMA, Toos Dekker-Nooren T, van Maanen K, Bode C F, Baird G, Kamp EM, 2000. A comparison of four serological tests for the diagnosis of caseous lymphadenitis in sheep and goats. Vet Microbiol, 75, 167-175.
  • Doherr MG, Carpenter TE, Wilson WD, Gardner IA, 1998. Application and evaluation of a mailed questionnaire for an epidemiologic study of Corynebacterium pseudotuberculosis infection in horses. Prev Vet Med, 35, 241-253.
  • Erganiş O, Kaya O, Ateş M, İstanbulluoglu E, 1990. Konya EBK kombinasında kesilen koyunlardaki apseli lenf yumruları üzerine mikrobiyolojik ve serolojik incelemeler. Veterinarium, 1, 8-11.
  • Esterabadi AH, Entessar F, Hedayati H, Narimani AA, Sadr IM, 1975. Isolation of Corynebacterium pseudotuberculosis from camel in Iran. Archives de L’Institut Razi, 27, 61-66. Fontaine MC, Baird GJ, 2008. Caseous lymphadenitis. Small Rum Res, 76, 42-48.
  • Funke G, Bernard, KA. 2003. Coryneform gram-positive rods, in: Manual of Clinical Microbiology, Eds: Murray PR, Baron EJ, Jorgensen JH, Pfaller MA, Yolken RH, ASM Press, Washington, USA, pp: 472-501.
  • Funke G, von Graevenitz A, Clarridge JE, Bernard KA, 1997. Clinical microbiology of coryneform bacteria. Clin Microbiol Rev, 10, 125-159.
  • Guimaraes AS, Seyffert N, Bastos BL, et al., 2009. Caseous lymphadenitis in sheep flocks of the state of Minas Gerais, Brazil: Prevalence and management surveys. Small Rum Res, 87, 86-91.
  • Hawari AD, 2008. Corynebacterium pseudotuberculosis infection (Caseous Lymphadenitis) in camels (Camelusdromedarius) in Jordan. Am J Anim Vet Sci, 3, 68-72.
  • Hodgson ALM, Tachedjian M, Corner LA, Radford AJ, 1994. Protection of sheep against caseous lymphadenitis by use of a single oral dose of live recombinant Corynebacterium pseudotuberculosis. Infect Immun, 62, 5275-5280.
  • Ilhan Z, 2013. Detection of Corynebacterium pseudotuberculosis from sheep lymph nodes by PCR. Revue Med Vet, 164, 2, 60-66.
  • Izgür M, Akan M, İlhan Z, 1999. Microorganisms isolated from cases of caseous lymphadenitis. Vet J Ankara Univ, 46, 43-50.
  • Judson R, Songer JG, 1991. Corynebacterium pseudotuberculosis: In-vitro susceptibility to 39 antimicrobial agents. Vet Microbiol, 27, 145-150.
  • Literak I, Horvathova A, Jahnova M, Rychlik L, Skalka B, 1999. Phenotype and genotype characteristics of the Slovak and Czech Corynebacterium pseudotuberculosis strains isolated from sheep and goats. Small Rum Res, 32, 107-111.
  • Lopez JF, Wong FM, Quesada J, 1966. Corynebacterium pseudotuberculosis: First case of human infection. Am J Clin Pathol, 46, 562-567.
  • Muckle CA, Gyles CL, 1982. Characterization of Strains of Corynebacterium pseudotuberculosis. Can J Comp Med, 46, 206-208.
  • NCCLS, 2003. National committee for clinical laboratory standards: Performance standards for antimicrobial susceptibility testing, 8th edition, NCCLS document M2-A8 volume 23, no 1, USA.
  • O’Reilly KM, Green LE, Malone FE, Medley GF, 2008. Parameter estimation and simulations of a mathematical model of Corynebacterium pseudotuberculosis transmission in sheep. Prev Vet Med, 83, 242-259.
  • Pacheco LGC, Pena RR, Castro TLP, et al., 2007. Multiplex PCR assay for identification of Corynebacterium pseudotuberculosis from pure cultures and for rapid detection of this pathogen in clinical samples. J Med Microbiol, 56, 480- 486.
  • Paton MW, Mercy AR, Sutherland SS, Ellis TM, 1988. The influence of shearing and age in the incidence of caseous lymphadenitis in Australian sheep flocks. Acta Vet Scand, 84, 101-103.
  • Paton MW, Walker SB, Rose IR, Watt GF, 2003. Prevalence of caseous lymphadenitis and usage of caseous lymphadenitis vaccines in sheep flocks. Aust Vet J, 81, 91-95.
  • Peel MM, Palmer GG, Stacpoole AM, Kerr TG, 1997. Human lymphadenitis due to Corynebacterium pseudotuberculosis: Report of ten cases from Australia and review. Clin Infect Dis, 24, 185-191.
  • Pepin M, Paton M and Hodgson AL, 1994. Pathogenesis and epidemiology of Corynebacterium pseudotuberculosis infection in sheep. Curr Topics Vet Res, 1, 63-82.
  • Prescott JF, Menzies PI, Hwang YT, 2002. An interferon-gamma assay for diagnosis of Corynebacterium pseudotuberculosis infection in adult sheep from a research flock. Vet Microbiol, 88, 287-297.
  • Quinn PJ, Carter ME, Markey BK, Carter GR, 1984. Clinical veterinary microbiology, Mosby International Ltd., Spain, pp: 118-345.
  • Seyffert N, Guimaraes AS, Pacheco LGC, et al., 2010. High seroprevalence of caseous lymphadenitis in Brazilian goat herds revealed by Corynebacterium pseudotuberculosis secreted proteins-based ELISA. Res Vet Sci, 88, 50-55.
  • Ter Laak EA, Bosch J, Bill GC, Schreuder BEC, 1992. Doubleantibody sandwich enzyme-linked immunosorbent assay and immunoblot analysis used for control of caseouslphadenitis in goats and sheep. Am J Vet Res, 53, 1125-1132.
  • Yeruham I, Elad D, Perl S, Ram A, 2003. Necrotic ulcerative dermatitis on the heels of heifers in a dairy herd infected with Corynebacterium pseudotuberculosis. Vet Rec, 152, 598-600.
  • Yeruham I, Elad D, van Ham M, Shpigel NY, Perl S, 1997. Corynebacterium pseudotuberculosis infection in Israeli cattle: Clinical and epidemiological studies. Vet Rec, 140, 423- 427.
There are 36 citations in total.

Details

Other ID JA59BR84PU
Journal Section Research
Authors

Aslı Sakmanoğlu This is me

Hasan Hüseyin Hadimli This is me

Osman Erganiş This is me

Yasemin Pınarkara This is me

Zafer Sayın This is me

Kürşat Kav This is me

Publication Date June 1, 2015
Published in Issue Year 2015 Volume: 31 Issue: 2

Cite

APA Sakmanoğlu, A., Hadimli, H. H., Erganiş, O., Pınarkara, Y., et al. (2015). Identification and antimicrobial susceptibility of Corynebacterium pseudotuberculosis isolated from sheep. Eurasian Journal of Veterinary Sciences, 31(2), 116-121.
AMA Sakmanoğlu A, Hadimli HH, Erganiş O, Pınarkara Y, Sayın Z, Kav K. Identification and antimicrobial susceptibility of Corynebacterium pseudotuberculosis isolated from sheep. Eurasian J Vet Sci. June 2015;31(2):116-121.
Chicago Sakmanoğlu, Aslı, Hasan Hüseyin Hadimli, Osman Erganiş, Yasemin Pınarkara, Zafer Sayın, and Kürşat Kav. “Identification and Antimicrobial Susceptibility of Corynebacterium Pseudotuberculosis Isolated from Sheep”. Eurasian Journal of Veterinary Sciences 31, no. 2 (June 2015): 116-21.
EndNote Sakmanoğlu A, Hadimli HH, Erganiş O, Pınarkara Y, Sayın Z, Kav K (June 1, 2015) Identification and antimicrobial susceptibility of Corynebacterium pseudotuberculosis isolated from sheep. Eurasian Journal of Veterinary Sciences 31 2 116–121.
IEEE A. Sakmanoğlu, H. H. Hadimli, O. Erganiş, Y. Pınarkara, Z. Sayın, and K. Kav, “Identification and antimicrobial susceptibility of Corynebacterium pseudotuberculosis isolated from sheep”, Eurasian J Vet Sci, vol. 31, no. 2, pp. 116–121, 2015.
ISNAD Sakmanoğlu, Aslı et al. “Identification and Antimicrobial Susceptibility of Corynebacterium Pseudotuberculosis Isolated from Sheep”. Eurasian Journal of Veterinary Sciences 31/2 (June 2015), 116-121.
JAMA Sakmanoğlu A, Hadimli HH, Erganiş O, Pınarkara Y, Sayın Z, Kav K. Identification and antimicrobial susceptibility of Corynebacterium pseudotuberculosis isolated from sheep. Eurasian J Vet Sci. 2015;31:116–121.
MLA Sakmanoğlu, Aslı et al. “Identification and Antimicrobial Susceptibility of Corynebacterium Pseudotuberculosis Isolated from Sheep”. Eurasian Journal of Veterinary Sciences, vol. 31, no. 2, 2015, pp. 116-21.
Vancouver Sakmanoğlu A, Hadimli HH, Erganiş O, Pınarkara Y, Sayın Z, Kav K. Identification and antimicrobial susceptibility of Corynebacterium pseudotuberculosis isolated from sheep. Eurasian J Vet Sci. 2015;31(2):116-21.