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Koyunlarda Kazeöz Lenfadenitisin Moleküler ve ELISA Yöntemiyle Karşılaştırmalı Teşhisi

Year 2022, Volume: 15 Issue: 2, 104 - 109, 31.12.2022
https://doi.org/10.47027/duvetfd.1192616

Abstract

Bu çalışmada; Diyarbakır yöresindeki koyunlarda Kazeöz Lenfadenitis’in (KLA) etiyolojik ajanı olan Corynebacterium. pseudotuberculosis (C. pseudotuberculosis)’in bakteriyolojik kültür, PCR ve ELISA teknikleri kullanılarak tanısının ortaya konulması, izolasyon bulguları ile testlerin özgüllük ve duyarlılıklarının belirlenmesi ve böylece hastalığın rutin teşhisinde kullanılabilirliklerinin ortaya konulması amaçlanmıştır. Çalışmanın materyalini Diyarbakır’ın farklı bölgelerinde olmak üzere 11 ayrı sürüden toplamda 2650 adet Akkaraman koyun oluşturdu. Sağlık taramasından geçirilen koyunların içinden 90 adet KLA semptomları gösteren ve yüzeysel lenf yumrularından bir veya birkaç tanesi birden apseleşmiş hasta koyunlar tespit edilerek çalışmaya dahil edildi. Kan örnekleri ve apse içeriğinden swap örnekleri alındı. Etkenin tanısı için, swap örneklerinden bakteriyolojik kültür ve PCR analizleri, kan örneklerinden ise ELISA analizleri yapıldı. Diyarbakır yöresindeki koyunlarda PCR analizlerine göre 26 (%28.9) pozitif, 64 (%71.1) negatif, ELISA analizinde 17 (%18.9) pozitif, 73 (%81.1) negatif ve bakteriyolojik kültürde 12 (%13.3)’sinde C. pseudotuberculosis, 16’sından Staphylococcus aureus (S. aureus), 8’inde Streptococ spp. ve 6’sında Escherichia coli (E. coli) kolonileri izole edilirken 48 (%53.3) kültürde hiç üreme olmadığı görüldü. PCR, ELISA ve bakteriyolojik kültür sonuçları karşılaştırıldığında PCR’ın daha belirleyici bir tanı testi olduğu görüldü (p<0.000). İzolasyon sonuçları dikkate alındığında KLA’nın tanısında PCR’ın ELISA’ya oranla duyarlılığı %100, özgüllüğü ise %65 olarak tespit edildi. Sonuç olarak KLA’nın bölge koyunlarında görüldüğü ve bu nedenle bölgede bu hastalığın önlenmesi için kontrol ve eradikasyon çalışmalarının yapılması gerektiği kanaatine varıldı.

Supporting Institution

DÜBAP

Project Number

veteriner.09

References

  • Fontaine MC, Baird GJ. (2008). Caseous Lymphadenitis. Small Rumin Res. 76 (1-2):42-48.
  • Sakmanoğlu A. (2015). Corynebacterium Pseudotuberculosis Fosfolipaz d (PLD) Geninin Aktarımı ve PLD Enziminin Aşı Olarak Kullanılması. Selçuk Üniversitesi Sağlık Bilimleri Enstitüsü, Doktora Tezi, Konya.
  • Al-Gaabary MH, SA Osman, AF Oreiby. (2009). Caseous Lymphadenitis in Sheep and Goats: Clinical, Epidemiological and Preventive Studies. Small Rumin Res. 87 (1-3): 116-121.
  • Dorella FA, Pacheco LGC, Oliveria SC, Miyoshi A, Azevedo V. (2006). Corynebacterium Pseudotuberculosis: Microbiology, Biochemical Properties, Pathogenesis and Molecular Studies of Virulence. Vet Res. 37(2): 201-218.
  • Baird GJ, Fontaine MC. (2007). Corynebacterium Pseudotuberculosis and Its Role in Ovine Caseous Lymphadenitis. J Comp Pathol. 137(4): 179-210.
  • LH Williamson. (2001). Caseous Lymphadenitis in Small Ruminants. Vet Clin North Am Food Anim.17(2):359-371.
  • Guimaraes A, do Carmo FB, Pauletti RB et al. (2011). Caseous Lymphadenitis: Epidemiology, Diagnosis, and Control. IIOAB J. 2(2): 33-43
  • Ali AD, Mahmoud AA, Khadr AM, Elshemey TM, Abdelrahman AH. (2016). Corynebacterium Pseudotuberculosis: Disease Prevalence, Lesion Distribution, and Diagnostic Comparison Through Microbiological Culture and Molecular Diagnosis. Alex J Vet Sci. 51(2): 189-198.
  • Umer M, Abba Y, Jesse F, Sharif A. (2017). Caseous Lymphadenitis in Small Ruminants: An Overview on Reproductive İmplications. J Vet Sci Anim Husb. 2(2): 23-31.
  • Batey RG. (1986). Pathogenesis of Caseous Lymphadenitis in Sheep and Goats. Aust Vet J. 63(9): 269-272
  • Osman AY, Nordin ML, Kadir AA, Saharee AA. (2018). The Epidemiology and Pathophysiology of Caseous Lymphadenitis: A review. J Vet Med Res. 5(3): 1129.
  • Ilhan, Z. (2003). Koyunlarda Corynebacterium Pseudotuberculosis' in ELISA ve Dot-Blot ELISA ile Teşhisi. Turkish J Vet Anim Sci. 27(6).
  • Stoops SG, Renshaw HW, Thilsted JP. (1984). Ovine Caseous Lymphadenitis: Disease Prevalence, Lesion Distribution, and Thoracic Manifestations in a Population of Mature Culled Sheep From Western United States. Am J Vet Res.45: 557-561.
  • Al-Rawashdeh OF, Al-Qudah KM. (2000). Effect of Shearing on the Incidence of Caseous Lymphadenitis in Awassi Sheep in Jordan. J Vet Med B Infect Dis Vet Public Health. 47: 287-293
  • Selim AM, Atwa SM, El Gedawy AA, Younis EE. (2021). Epidemiological, Bacteriological and Molecular Studies on Caseous Lymphadenitis in Sheep of Dakhlia, Egypt. J Anim Sci Biotechnol.1-6.
  • Cetinkaya B, Karahan M, Atil E. (2002). Identification of Corynebacterium Pseudotuberculosis Isolates From Sheep and Goats by PCR. Vet Microbiol. 88(1): 75-83.
  • Aslan Ö, Gümüşsoy KS, Bekdik İK, Akcay A,Demiral ÖO. (2016). Seroprevalence of Caseous Lymphadenitis in Kangal Akkaraman Sheep. Turkish J Vet Anim Sci. 40(6): 811-816.
  • Muz A, Eroksuz H, Ongor H, et al. (1995). Microbiological, Serological and Pathological Investigations on The Prescapular Lymph Nodes with Abscesses From Sheep in the Abattoir of Meat and Fish Organisation in Elazig. FU Saglik Bil Derg. 9, 204–211
  • Alves JRA, de Farias AEM, da Silva JD, et al. (2020). Factors Associated with the Seroprevalence of Caseous Lymphadenitis in Sheep from Northeastern Brazil. Prev Vet Med. 182, 105098.
  • Seyffert N, Guimarães AS, Pacheco LG. 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.
  • Farias AM, Alves JR, Alves FS. et al. (2018). Serological Study on Corynebacterium Pseudotuberculosis Infection in Goats in the Brazilian Northeast Using Enzyme-Linked Immunosorbent Assay (ELISA-indirect). Pesq Vet Bras. 38: 1344-1350
  • Costa L, Huerta B, Galán‐Relaño Á, et al. (2020). Utility Assessment of an Enzyme‐Linked Immunosorbent Assay for Detection of Subclinical Cases of Caseous Lymphadenitis in Small Ruminant Flocks. Vet Med Sci. 6(4): 796-803.
  • Tripathi BN, Kumar J. Sonawane GG, Kumar R, Dixit SK. (2016). Microbiological and Molecular Investigation of Clinically Suspected Caseous Lymphadenitis Cases in Goats. Agric Res. 5(4): 413-419.
  • Kumar J, Tripathi BN, Kumar R, Sonawane GG, Dixit SK. (2013). Rapid Detection of Corynebacterium Pseudotuberculosis in Clinical Samples From Sheep. Trop Anim Health and Prod.45(6): 1429-1435.
  • Chirino-Zárraga C, Scaramelli A, Rey-Valeirón C. (2006). Bacteriological Characterization of Corynebacterium Pseudotuberculosis in Venezuelan Goat flocks. Small Rumin Res. 65, 170-175.
  • Mubarak M, Bastawrows AF, Abdel-Hafeez MM, Ali MM. (1999). Caseous Lymphadenitis of Sheep and Goats in Assiut Farms and Abattoirs. Assiut Vet Med J. 42: 89-112
  • Guimaraes AS, Carmo FB, Heinemann MB, et al. (2011). High Sero-prevalence of Caseous Lymphadenitis Identified in Slaughterhouse Samples as a Consequence of Deficiencies in Sheep Farm Management in the State of Minas Gerais, Brazil. BMC Vet Res. 7: 68.
  • Costa L, Maldonado A, Huerta B, Almeida A. (2019). Optimization of a Conventional PCR Assay for the Identification of Corynebacterium Pseudotuberculosis from Pyogenic Lesions. Int J Vet Sci Anim Husb. 7(2): 201-210
  • Nassar AFD, Daniel GT, Ruiz R, et al. (2016). Diagnostic comparison of Corynebacterium Pseudotuberculosis Through Microbiological Kulture and PCR in Sheep Samples. Arq Inst Biol. 82: 1-6.
  • Jung BY, Lee SH, Kim HY, et al. (2015). Serology and Clinical Relevance of Corynebacterium Pseudotuberculosis in Native Korean Goats (Capra hircus coreanae). Trop Anim Health Prod. 47(4): 657-661.
  • Zavoshti FR, Khoojine ABS, Helan JA, Hassanzadeh B, Heydari AA. (2012). Frequency of Caseous Lymphadenitis (CLA) in Sheep Slaughtered in an Abattoir in Tabriz: Comparison of bacterial culture and pathological study. Comp Clin Path. 21(5): 667-671.
  • Chikhaoui M, Khoudja FB. (2013). Clinicopathological Investigation on Caseous Lymphadenitis in Local Breed Sheep in Algeria. Trop Anim Health Prod. 45(7): 1641-1643
  • Parin U, Kirkan S, Ural K, et al. (2018). Molecular Identification of Corynebacterium Pseudotuberculosis in sheep. Acta Veterinaria Brno. 87(1): 3-8.

Comparative Diagnosis of Caseous Lymphadenitis in Sheep by Molecular and ELISA Method

Year 2022, Volume: 15 Issue: 2, 104 - 109, 31.12.2022
https://doi.org/10.47027/duvetfd.1192616

Abstract

In this study; It is aimed to reveal the diagnosis of Corynebacterium. pseudotuberculosis (C. pseudotuberculosis), which is the etiological agent of Caseous Lymphadenitis (CLA), in sheep in the Diyarbakir region using bacteriological culture, PCR and ELISA techniques, to determine the specificity and sensitivity of the tests with isolation findings, and thus to demonstrate their usability in the routine diagnosis of the disease. The material of the study consisted of 2650 Akkaraman sheep from 11 different herds in different regions of Diyarbakır. Among the sheep undergoing health screening, 90 sick sheep with CLA symptoms and one or more of the superficial lymph nodes with abscess were identified and included in the study. Blood samples and swap samples were taken from the abscess contents. Bacteriological culture and PCR analyzes were performed on swap samples and ELISA analyzes were performed on blood samples for the diagnosis of the causative agent. According to PCR analysis, 26 (28.9%) positive, 64 (71.1%) negative, 17 (18.9%) positive ELISA analysis, 73 (81.1%) negative, and 12 (13.3%) bacteriological cultures were found in sheep in Diyarbakir region. While C. pseudotuberculosis was isolated in 12, Staphylococcus aureus (S. aureus) in 16, Streptococ spp in 8, and Escherichia coli (E. coli) in 6, no growth was observed in 48 (53.3%) cultures appeared not to be. When PCR, ELISA and bacteriological culture results were compared, PCR was found to be a more definitive diagnostic test (p<0.000). Considering the isolation results, the sensitivity of PCR compared to ELISA was 100% and the specificity was 65%. As a result, it was concluded that CLA is seen in the sheep of the region and therefore, control and eradication studies should be carried out in order to prevent this disease in the region.

Project Number

veteriner.09

References

  • Fontaine MC, Baird GJ. (2008). Caseous Lymphadenitis. Small Rumin Res. 76 (1-2):42-48.
  • Sakmanoğlu A. (2015). Corynebacterium Pseudotuberculosis Fosfolipaz d (PLD) Geninin Aktarımı ve PLD Enziminin Aşı Olarak Kullanılması. Selçuk Üniversitesi Sağlık Bilimleri Enstitüsü, Doktora Tezi, Konya.
  • Al-Gaabary MH, SA Osman, AF Oreiby. (2009). Caseous Lymphadenitis in Sheep and Goats: Clinical, Epidemiological and Preventive Studies. Small Rumin Res. 87 (1-3): 116-121.
  • Dorella FA, Pacheco LGC, Oliveria SC, Miyoshi A, Azevedo V. (2006). Corynebacterium Pseudotuberculosis: Microbiology, Biochemical Properties, Pathogenesis and Molecular Studies of Virulence. Vet Res. 37(2): 201-218.
  • Baird GJ, Fontaine MC. (2007). Corynebacterium Pseudotuberculosis and Its Role in Ovine Caseous Lymphadenitis. J Comp Pathol. 137(4): 179-210.
  • LH Williamson. (2001). Caseous Lymphadenitis in Small Ruminants. Vet Clin North Am Food Anim.17(2):359-371.
  • Guimaraes A, do Carmo FB, Pauletti RB et al. (2011). Caseous Lymphadenitis: Epidemiology, Diagnosis, and Control. IIOAB J. 2(2): 33-43
  • Ali AD, Mahmoud AA, Khadr AM, Elshemey TM, Abdelrahman AH. (2016). Corynebacterium Pseudotuberculosis: Disease Prevalence, Lesion Distribution, and Diagnostic Comparison Through Microbiological Culture and Molecular Diagnosis. Alex J Vet Sci. 51(2): 189-198.
  • Umer M, Abba Y, Jesse F, Sharif A. (2017). Caseous Lymphadenitis in Small Ruminants: An Overview on Reproductive İmplications. J Vet Sci Anim Husb. 2(2): 23-31.
  • Batey RG. (1986). Pathogenesis of Caseous Lymphadenitis in Sheep and Goats. Aust Vet J. 63(9): 269-272
  • Osman AY, Nordin ML, Kadir AA, Saharee AA. (2018). The Epidemiology and Pathophysiology of Caseous Lymphadenitis: A review. J Vet Med Res. 5(3): 1129.
  • Ilhan, Z. (2003). Koyunlarda Corynebacterium Pseudotuberculosis' in ELISA ve Dot-Blot ELISA ile Teşhisi. Turkish J Vet Anim Sci. 27(6).
  • Stoops SG, Renshaw HW, Thilsted JP. (1984). Ovine Caseous Lymphadenitis: Disease Prevalence, Lesion Distribution, and Thoracic Manifestations in a Population of Mature Culled Sheep From Western United States. Am J Vet Res.45: 557-561.
  • Al-Rawashdeh OF, Al-Qudah KM. (2000). Effect of Shearing on the Incidence of Caseous Lymphadenitis in Awassi Sheep in Jordan. J Vet Med B Infect Dis Vet Public Health. 47: 287-293
  • Selim AM, Atwa SM, El Gedawy AA, Younis EE. (2021). Epidemiological, Bacteriological and Molecular Studies on Caseous Lymphadenitis in Sheep of Dakhlia, Egypt. J Anim Sci Biotechnol.1-6.
  • Cetinkaya B, Karahan M, Atil E. (2002). Identification of Corynebacterium Pseudotuberculosis Isolates From Sheep and Goats by PCR. Vet Microbiol. 88(1): 75-83.
  • Aslan Ö, Gümüşsoy KS, Bekdik İK, Akcay A,Demiral ÖO. (2016). Seroprevalence of Caseous Lymphadenitis in Kangal Akkaraman Sheep. Turkish J Vet Anim Sci. 40(6): 811-816.
  • Muz A, Eroksuz H, Ongor H, et al. (1995). Microbiological, Serological and Pathological Investigations on The Prescapular Lymph Nodes with Abscesses From Sheep in the Abattoir of Meat and Fish Organisation in Elazig. FU Saglik Bil Derg. 9, 204–211
  • Alves JRA, de Farias AEM, da Silva JD, et al. (2020). Factors Associated with the Seroprevalence of Caseous Lymphadenitis in Sheep from Northeastern Brazil. Prev Vet Med. 182, 105098.
  • Seyffert N, Guimarães AS, Pacheco LG. 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.
  • Farias AM, Alves JR, Alves FS. et al. (2018). Serological Study on Corynebacterium Pseudotuberculosis Infection in Goats in the Brazilian Northeast Using Enzyme-Linked Immunosorbent Assay (ELISA-indirect). Pesq Vet Bras. 38: 1344-1350
  • Costa L, Huerta B, Galán‐Relaño Á, et al. (2020). Utility Assessment of an Enzyme‐Linked Immunosorbent Assay for Detection of Subclinical Cases of Caseous Lymphadenitis in Small Ruminant Flocks. Vet Med Sci. 6(4): 796-803.
  • Tripathi BN, Kumar J. Sonawane GG, Kumar R, Dixit SK. (2016). Microbiological and Molecular Investigation of Clinically Suspected Caseous Lymphadenitis Cases in Goats. Agric Res. 5(4): 413-419.
  • Kumar J, Tripathi BN, Kumar R, Sonawane GG, Dixit SK. (2013). Rapid Detection of Corynebacterium Pseudotuberculosis in Clinical Samples From Sheep. Trop Anim Health and Prod.45(6): 1429-1435.
  • Chirino-Zárraga C, Scaramelli A, Rey-Valeirón C. (2006). Bacteriological Characterization of Corynebacterium Pseudotuberculosis in Venezuelan Goat flocks. Small Rumin Res. 65, 170-175.
  • Mubarak M, Bastawrows AF, Abdel-Hafeez MM, Ali MM. (1999). Caseous Lymphadenitis of Sheep and Goats in Assiut Farms and Abattoirs. Assiut Vet Med J. 42: 89-112
  • Guimaraes AS, Carmo FB, Heinemann MB, et al. (2011). High Sero-prevalence of Caseous Lymphadenitis Identified in Slaughterhouse Samples as a Consequence of Deficiencies in Sheep Farm Management in the State of Minas Gerais, Brazil. BMC Vet Res. 7: 68.
  • Costa L, Maldonado A, Huerta B, Almeida A. (2019). Optimization of a Conventional PCR Assay for the Identification of Corynebacterium Pseudotuberculosis from Pyogenic Lesions. Int J Vet Sci Anim Husb. 7(2): 201-210
  • Nassar AFD, Daniel GT, Ruiz R, et al. (2016). Diagnostic comparison of Corynebacterium Pseudotuberculosis Through Microbiological Kulture and PCR in Sheep Samples. Arq Inst Biol. 82: 1-6.
  • Jung BY, Lee SH, Kim HY, et al. (2015). Serology and Clinical Relevance of Corynebacterium Pseudotuberculosis in Native Korean Goats (Capra hircus coreanae). Trop Anim Health Prod. 47(4): 657-661.
  • Zavoshti FR, Khoojine ABS, Helan JA, Hassanzadeh B, Heydari AA. (2012). Frequency of Caseous Lymphadenitis (CLA) in Sheep Slaughtered in an Abattoir in Tabriz: Comparison of bacterial culture and pathological study. Comp Clin Path. 21(5): 667-671.
  • Chikhaoui M, Khoudja FB. (2013). Clinicopathological Investigation on Caseous Lymphadenitis in Local Breed Sheep in Algeria. Trop Anim Health Prod. 45(7): 1641-1643
  • Parin U, Kirkan S, Ural K, et al. (2018). Molecular Identification of Corynebacterium Pseudotuberculosis in sheep. Acta Veterinaria Brno. 87(1): 3-8.
There are 33 citations in total.

Details

Primary Language Turkish
Subjects Veterinary Surgery
Journal Section Research
Authors

Ayşe Ekinci Yıldız 0000-0003-4590-8218

Hasan İçen 0000-0002-6034-3203

Project Number veteriner.09
Publication Date December 31, 2022
Acceptance Date December 20, 2022
Published in Issue Year 2022 Volume: 15 Issue: 2

Cite

APA Ekinci Yıldız, A., & İçen, H. (2022). Koyunlarda Kazeöz Lenfadenitisin Moleküler ve ELISA Yöntemiyle Karşılaştırmalı Teşhisi. Dicle Üniversitesi Veteriner Fakültesi Dergisi, 15(2), 104-109. https://doi.org/10.47027/duvetfd.1192616
AMA Ekinci Yıldız A, İçen H. Koyunlarda Kazeöz Lenfadenitisin Moleküler ve ELISA Yöntemiyle Karşılaştırmalı Teşhisi. Dicle Üniv Vet Fak Derg. December 2022;15(2):104-109. doi:10.47027/duvetfd.1192616
Chicago Ekinci Yıldız, Ayşe, and Hasan İçen. “Koyunlarda Kazeöz Lenfadenitisin Moleküler Ve ELISA Yöntemiyle Karşılaştırmalı Teşhisi”. Dicle Üniversitesi Veteriner Fakültesi Dergisi 15, no. 2 (December 2022): 104-9. https://doi.org/10.47027/duvetfd.1192616.
EndNote Ekinci Yıldız A, İçen H (December 1, 2022) Koyunlarda Kazeöz Lenfadenitisin Moleküler ve ELISA Yöntemiyle Karşılaştırmalı Teşhisi. Dicle Üniversitesi Veteriner Fakültesi Dergisi 15 2 104–109.
IEEE A. Ekinci Yıldız and H. İçen, “Koyunlarda Kazeöz Lenfadenitisin Moleküler ve ELISA Yöntemiyle Karşılaştırmalı Teşhisi”, Dicle Üniv Vet Fak Derg, vol. 15, no. 2, pp. 104–109, 2022, doi: 10.47027/duvetfd.1192616.
ISNAD Ekinci Yıldız, Ayşe - İçen, Hasan. “Koyunlarda Kazeöz Lenfadenitisin Moleküler Ve ELISA Yöntemiyle Karşılaştırmalı Teşhisi”. Dicle Üniversitesi Veteriner Fakültesi Dergisi 15/2 (December 2022), 104-109. https://doi.org/10.47027/duvetfd.1192616.
JAMA Ekinci Yıldız A, İçen H. Koyunlarda Kazeöz Lenfadenitisin Moleküler ve ELISA Yöntemiyle Karşılaştırmalı Teşhisi. Dicle Üniv Vet Fak Derg. 2022;15:104–109.
MLA Ekinci Yıldız, Ayşe and Hasan İçen. “Koyunlarda Kazeöz Lenfadenitisin Moleküler Ve ELISA Yöntemiyle Karşılaştırmalı Teşhisi”. Dicle Üniversitesi Veteriner Fakültesi Dergisi, vol. 15, no. 2, 2022, pp. 104-9, doi:10.47027/duvetfd.1192616.
Vancouver Ekinci Yıldız A, İçen H. Koyunlarda Kazeöz Lenfadenitisin Moleküler ve ELISA Yöntemiyle Karşılaştırmalı Teşhisi. Dicle Üniv Vet Fak Derg. 2022;15(2):104-9.