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Calicivirus ile enfekte kedilerde adenozin deaminaz 1, paraoksonaz 1, C-reaktif protein ve serum amiloid A düzeylerinin araştırılması

Yıl 2021, Cilt: 6 Sayı: 3, 133 - 138, 31.12.2021
https://doi.org/10.24880/maeuvfd.971635

Öz

Bu çalışma ile feline calicivirus (FCV) ile enfekte kedilerde hücresel immün yanıtın, oksidatif stresin ve yangısal sürecin belirlenmesi amaçlanmıştır. Çalışmada FCV ile enfekte 20 adet kedi ve 10 adet klinik olarak sağlıklı olmak üzere toplam 30 kedi kullanılmıştır. Çalışmada tüm kedilerden toplanan serum örneklerinde kedi spesifik ELISA test kitleri kullanılarak adenozin deaminaz 1 (ADA-1), paraoksonaz 1 (PON-1), C-reaktif protein (CRP) ve serum amiloid A (SAA) düzeyleri belirlenmiştir. Sonuç olarak, FCV ile enfekte kedilerin serum ADA-1, SAA ve CRP düzeyleri kontrol grubununkilerden istatistiksel olarak önemli düzeyde yüksek bulunmuştur. Ayrıca enfekte kedilerin PON-1 düzeyleri ise kontrol grubundan önemli düzeyde düşük olduğu belirlenmiştir (p<0,001). Yapılan bu çalışmada, ADA-1 ile PON-1 arasında yüksek düzeyde anlamlı negatif korelasyon (r=-0,73; p<0,001) saptanırken diğer parametreler arasında ise orta düzeyde korelasyonlar belirlenmiştir. Sonuç olarak, elde edilen veriler FCV ile enfekte kedilerde ADA-1 düzeyindeki artış aktive olmuş hücresel yanıtı, PON-1 düzeyindeki düşüş oksidatif stresin geliştiğini göstermektedir. Ayrıca SAA ve CRP düzeylerindeki artışlar ise bu kedilerde akut faz yanıtın geliştiğini ortaya koymaktadır. Bu çalışma, FCV ile enfekte kedilerde ADA-1’in hücresel immün yanıtın durumunun belirlenmesinde, PON-1’in oksidatif stresin belirlenmesinde yararlı biyomarkırlar olabilir. Bunlara ek olarak SAA ve CRP’nin gelişen yangıya bağlı akut faz yanıtın belirlenmesinde kullanılabilir.

Destekleyen Kurum

Mehmet Akif Ersoy Üniversitesi, Bilimsel araştırma projeleri koordinatörlüğü

Proje Numarası

0577-YL-19

Teşekkür

Mehmet Akif Ersoy Üniversitesi, Bilimsel araştırma projeleri kordinatörlüğü'ne projeyi desteklediği için teşekkür ederiz.

Kaynakça

  • 1- Akhtardanesh B, Ghalekhani, N, Abshenas J, Nematollahi H, Sharifi H. Serum adenosine deaminase as a diagnostic marker of chronic infectious disease in dogs. Vet Res 2013;17:592-595.
  • 2- Antonioli L, Colucci R, La Motta C, Tuccori M, Awwad O, Da Settimo F, Blandizzi C, Fornai M. Adenosine deaminase in the modulation of immüne system and its potential as a novel target for treatment of inflammatory disorders. Current Drug Targets, 2012;13:842-862.
  • 3- Baba K, Hoosen AA, Langeland N, Dyrhol-Riise AM. Adenosine Deaminase Activity Is a Sensitive Marker for the Diagnosis of Tuberculous Pleuritis in Patients with Very Low CD4 Counts. PLOSone, 208;3:1-5.
  • 4- Berger A, Willi B, Meli ML, Boretti FS, Hartnack S, Dreyfus A et al. Feline Calicivirus and other respiratory pathogens in cats with Feline Calicivirus related symptoms and in clinically healthy cats in Switzerland. Vet Res 2015;11:1-12.
  • 5- Brigida I, Sauer A, Ferrua F, Gianelli S, Scaramuzza S, Pistoia V et al. B-cell development and functions and therapeutic options in adenosine deaminase–deficient patients. J. Allergy and Clin Immunol, 2014;133:799–806.
  • 6- Ceron JJ, Tecles F, Tvarijonaviciute A. Serum paraoxonase 1 (PON1) measurement: an update. BMC Vet Res 2014;10:1-11.
  • 7- Chiu IM, Huang LC, Chen IL, Tang KS, Huang YH. Diagnostic values of C-reactive protein and complete blood cell to identify invasive bacterial infection in young febrile infants. Pediatrics and Neonatology 2019;60:197-200.
  • 8- Climent N, Martinez-Navio JM, Gil C, Garcia F, Rovia C, Hurtado C et al. Adenosine deaminase enhances T-cell response elicited by dendritic cells loaded with inactivated HIV. Immunol and Cell Biology 2009; 87:634–639.
  • 9- Coyne KP, Jones BRD, Kipar A, Chantrey J, Porter CJ, Barber PJ et al. Lethal outbreak of a disease associated with feline Calicivirus infection in cats. Vet Record, 2006a;158:544-550.
  • 10- Coyne KP, Dawson S, Radford AD, Cripps PJ, Porter CJ, McCracken CM et al. Long term analysis of feline Calicivirus prevalence and viral shedding patterns in naturally infected colonies of domestic cats. Vet Microbiol 2006b;118(1-2):12-25.
  • 11- Coyne KP, Gaskell RM, Dawson S, Porter CJ, Radford AD. Evolutionary mechanisms of persistence and diversification of a Calicivirus within endemically infected natural host populations. J Virology, 2007;81(4):1961-1971.
  • 12- Çırak KA, Tekgül S, Bilaçeroğlu S, Kömürcü B, Uçar MA, Yalnız E. Diagnostic values of pleural fluid, and serum C-reactive protein, Adenosine deaminase, and Lactate dehydrogenase levels and pleural fluid/serum ratios in the differentiation of malignant from benign pleural effusions. İzmir Göğüs Hastanesi Dergisi, 2012;2:75-82.
  • 13- Dowers KL, Hawley JR, Brewer MM, Morris AK, Radecki SV, Lappin MR. Association of Bartonella species, feline Calicivirus, and feline herpesvirus 1 infection with gingivostomatitis in cats. J Feline Med Surg, 2010;12(4):314-321.
  • 14- Ellah MRABD, Nishimori K, Goryo M, Okada K, Yasuda J. Serum adenosine deaminase activity in bovine liver diseases, J Vet Med Sci, 2004;66(11):1421-1422.
  • 15- Farid AS, Kazuyuki Honkawa K, Fath EM, Nonaka N, Horii Y. Serum paraoxonase-1 as biomarker for improved diagnosis of fatty liver in dairy cows. BMC Vet Res, 2013;9:1-11.
  • 16- Flinn AM, Gennery A. Adenosine deaminase deficiency: a review. Orphanet J Rare Dis 2018;13:1-7.
  • 17- Foley J, Hurley K, Pesavento PA, Poland A, Pedersen NC. Virulent systemic feline Calicivirus infection: local cytokine modulation and contribution of viral mutants. J Feline Med Surg 2006;8:55-61.
  • 18- Gökce Hİ ve Bozukluhan K. Çiftlik Hayvanlarında Önemli Akut Faz Proteinleri ve Bunların Veteriner Hekimlikte Alanındaki kullanımı. Dicle Üniv Vet Fak Derg 2009;1(1):1-14.
  • 19- Kahraman MA, Gökce HI. Investigations of Adenosine Deaminase and C-reactive Protein in Cats with Feline Infectious Peritonitis. MAE J Health Sci Inst., 2020; 8,98-107.
  • 20- Kim JW, Yang IA, Oh AE, Rhyhoo YG, Jang YH . C reactive protein, sialic acid and adenosine deaminase levels in serum and pleural fluid from patients with pleural effusion. The Korean J Int Med 1988;3:122-127.
  • 21- Kodama J, Miyagi Y, Seki N, Tokumo K, Yoshinouchi M, Kobashi Y et al. Serum C-reactive protein as a prognostic factor in patients with epithelial ovarian cancer. European Journal of . Obstetrics&Gynecology 1999;82:107.
  • 22- Kulka M. A review of paraoxonase 1 properties and diagnostic applications. Polish J Vet Sci 2016;19:225–232.
  • 23- Litvinov D, Mahini H, Garelnabi M. Antioxidant and Anti-Inflammatory Role of Paraoxonase 1: Implication in Arteriosclerosis Diseases. North Am J Med&Sci 2012;4(11):523–532.
  • 24- Meazzi S, Ferriani R, Paltrinieri S, Giordano A Preliminary data about Paraoxonase-1 (PON-1) as a maker for Feline Infectious Peritonitis (FIP). Int J Health, Anim Sci & Food Safety, 2018;5(1):1-5.
  • 25- Novak F, Vavrova L, Kodydkova J, Novak F Sr, Hynkova M, Zak A et al. Decreased paraoxonase activity in critically ill patients with sepsis. Clin Exp Med 2010;10:21–25.
  • 26- Poulet H, Jas D, Lemeter C, Coupier C, Brunet S. Efficacy of a bivalent inactivated non-adjuvanted feline calicivirus vaccine: Relation between in vitro cross-neutralization and heterologous protection in vivo. Vaccine 2008;26:3647–3654.
  • 27- Radford AD, Addie D, Belák S, Boucraut-Baralon C, Egberink H, Frymus T et al. Feline Calicivirus infection. ABCD guidelines on prevention and management. J Feline Med Surg 2009;11(7):556-64.
  • 28- Reynolds BS, Poulet H, Pingret JL, Jas D, Brunet S, Lemeter C et al. A nosocomial outbreak of feline Calicivirus associated virulent systemic disease in France. J Feline Med Surg 2009;11(8):633-644.
  • 29- Rodrigues DSLF, Oliveira, MEF, Teixeira PPM, Cavalcante IJM, Vale MR). Adenosine deaminase activity as a biochemical marker of inflammatory response in goats infected by caprine arthritis–encephalitis virus. Small Rum Res 2012;108(1–3):120-126.
  • 30- Rossi G, Giordano A, Pezzia F, Kjelgaard-Hansen M, Paltrin S. Serum paraoxonase 1 activity in dogs: Preanalytical and analytical factors and correlation with C-reactive protein and alpha-2-globulin. Vet Clin Pathol, 2013;42:329-341.
  • 31- Ruiz-Gonzalez A, Bielsa S, Falguera M, Porcel JM. The Diagnostic Value of Serum C-Reactive Protein for Identifying Pneumonia in Hospitalized Patients with Acute Respiratory Symptoms. J Biomarkers, 2016;2016:1-5.
  • 32- Scavone D, Sgorbini M, Borges AS, Oliveira-Filho JP, Vitale V, Paltrinieri S. Serial measurements of Paraoxonase-1 (PON-1) activity in horses with experimentally induced endotoxemia. BMC Vet Res 2020;16:1-7.
  • 33- Schulz C, Hartmann K, Mueller RS, Helps C, Schulz BS. Sampling sites for detection of feline herpesvirus-1, feline Calicivirus and Chlamydia felis in cats with feline upper respiratory tract disease, J Feline Med Surg 2015;17(12):1012-9.
  • 34- Shunmoogam N, Naidoo P, Chilton R. Paraoxonase (PON)-1: a brief overview on genetics, structure, polymorphisms and clinical relevance. Vascular Health and Risk Management, 2018;14:137–143.
  • 35- Sproston NR, Ashworth JJ. Role of C-Reactive Protein at Sites of inflammation and infection. Frontiers in Imm

Investigations of adenosine deaminase 1, paraoxonase 1, C-reactive protein and serum amyloid A levels in cats infected with calicivirus. (Calicivirus ile enfekte kedilerde adenozin deaminaz 1, paraoksonaz 1, C-reaktif protein ve serum amiloid A düzeylerinin araştırılması)

Yıl 2021, Cilt: 6 Sayı: 3, 133 - 138, 31.12.2021
https://doi.org/10.24880/maeuvfd.971635

Öz

ÖZ
Bu çalışma ile feline calicivirus (FCV) ile enfekte kedilerde hücresel immün yanıtın, oksidatif stresin ve yangısal sürecin belirlenmesi amaçlanmıştır. Çalışmada FCV ile enfekte 20 adet kedi ve 10 adet klinik olarak sağlıklı kedi kullanılmıştır. Çalışmada tüm kedilerden toplanan serum örneklerinde kedi spesifik ELISA test kitleri kullanılarak adenozin deaminaz 1 (ADA-1), paraoksonaz 1 (PON-1), C-reaktif protein (CRP) ve serum amiloid A (SAA) düzeyleri belirlenmiştir. Yapılan analizler sonucunda FCV ile enfekte kedilerin serum ADA-1, SAA ve CRP düzeyleri kontrol grubununkilerden istatistiksel olarak önemli düzeyde yüksek bulunmuştur. Ayrıca enfekte kedilerin PON-1 düzeyleri ise kontrol grubudan önemli düzeyde düşük olduğu belirlenmiştir (p<0,001). Yapılan bu çalışmada, ADA-1 ile PON-1 arasında yüksek düzeyde anlamlı negatif korelasyon (r=-0,73; p<0,001) saptanırken diğer parametreler arasında ise orta düzeyde korelasyonlar belirlenmiştir. Elde edilen veriler FCV ile enfekte kedilerde ADA-1 düzeyindeki artış aktive olmuş hücresel yanıtı, PON-1 düzeyindeki düşüş oksidatif stresin geliştiğini, SAA ve CRP düzeylerindeki artışlar ise bu kedilerde akut faz yanıtın geliştiğini ortaya koymaktadır. Sonuç olarak, bu çalışma ile FCV ile enfekte kedilerde ADA-1’in hücresel immün yanıtın durumunun belirlenmesinde, PON-1’in oksidatif stres ve antioksidan kapasitenin belirlenmesinde ve ayrıca SAA ve CRP’nin gelişen yangıya bağlı akut faz yanıtın belirlenmesinde yararlı biyomarkırlar olarak kullanılabileceği ortaya konulmuştur.
Anahtar Kelimeler: Biyokimya, Calicivirus, Kedi

ABSTRACT
Abstract: The aims of this study were determination of cellular immune responses, oxidative stress, and inflammatory responses in cats infected with FCV. In the study, 20 cats infected with FCV and 10 clinically healthy cats were used. In the study, adenosine deaminase 1 (ADA-1), paraoxonase 1 (PON-1), C reactive protein (CRP) and serum amyloid A (SAA) levels were determined in collected serum samples of all the cats, using cat-specific ELISA test kits. In the present study, serum concentrations of ADA-1, SAA and CRP were significantly higher than those of control group (p< 0,001). Wheras PON-1 concentrations were found to be significantly low compared to that of control group. Furthermore, high negative correlations were obtained between ADA-1 and PON-1 (r=-0,73; p<0,001), while modarete correlations were obtained between other parameters. In this study, elevated serum ADA-1 levels indicate activated cellular immune responses and decreased PON-1 levels indicate oxidative stress. Furthermore, increased SAA and CRP levels determine the development of acute phase responses in cats infected with FCV. In conclusion, ADA-1 and PON-1 can be useful biomarkers for detecting activated cellular immune responses and oxidative stress in cats infected with FCV, respectively. Furthermore, SAA and CRP can be used to detect acute phase responses in infected cats with FCV.
Key words: Biochemistry, Calicivirus, Cat

Proje Numarası

0577-YL-19

Kaynakça

  • 1- Akhtardanesh B, Ghalekhani, N, Abshenas J, Nematollahi H, Sharifi H. Serum adenosine deaminase as a diagnostic marker of chronic infectious disease in dogs. Vet Res 2013;17:592-595.
  • 2- Antonioli L, Colucci R, La Motta C, Tuccori M, Awwad O, Da Settimo F, Blandizzi C, Fornai M. Adenosine deaminase in the modulation of immüne system and its potential as a novel target for treatment of inflammatory disorders. Current Drug Targets, 2012;13:842-862.
  • 3- Baba K, Hoosen AA, Langeland N, Dyrhol-Riise AM. Adenosine Deaminase Activity Is a Sensitive Marker for the Diagnosis of Tuberculous Pleuritis in Patients with Very Low CD4 Counts. PLOSone, 208;3:1-5.
  • 4- Berger A, Willi B, Meli ML, Boretti FS, Hartnack S, Dreyfus A et al. Feline Calicivirus and other respiratory pathogens in cats with Feline Calicivirus related symptoms and in clinically healthy cats in Switzerland. Vet Res 2015;11:1-12.
  • 5- Brigida I, Sauer A, Ferrua F, Gianelli S, Scaramuzza S, Pistoia V et al. B-cell development and functions and therapeutic options in adenosine deaminase–deficient patients. J. Allergy and Clin Immunol, 2014;133:799–806.
  • 6- Ceron JJ, Tecles F, Tvarijonaviciute A. Serum paraoxonase 1 (PON1) measurement: an update. BMC Vet Res 2014;10:1-11.
  • 7- Chiu IM, Huang LC, Chen IL, Tang KS, Huang YH. Diagnostic values of C-reactive protein and complete blood cell to identify invasive bacterial infection in young febrile infants. Pediatrics and Neonatology 2019;60:197-200.
  • 8- Climent N, Martinez-Navio JM, Gil C, Garcia F, Rovia C, Hurtado C et al. Adenosine deaminase enhances T-cell response elicited by dendritic cells loaded with inactivated HIV. Immunol and Cell Biology 2009; 87:634–639.
  • 9- Coyne KP, Jones BRD, Kipar A, Chantrey J, Porter CJ, Barber PJ et al. Lethal outbreak of a disease associated with feline Calicivirus infection in cats. Vet Record, 2006a;158:544-550.
  • 10- Coyne KP, Dawson S, Radford AD, Cripps PJ, Porter CJ, McCracken CM et al. Long term analysis of feline Calicivirus prevalence and viral shedding patterns in naturally infected colonies of domestic cats. Vet Microbiol 2006b;118(1-2):12-25.
  • 11- Coyne KP, Gaskell RM, Dawson S, Porter CJ, Radford AD. Evolutionary mechanisms of persistence and diversification of a Calicivirus within endemically infected natural host populations. J Virology, 2007;81(4):1961-1971.
  • 12- Çırak KA, Tekgül S, Bilaçeroğlu S, Kömürcü B, Uçar MA, Yalnız E. Diagnostic values of pleural fluid, and serum C-reactive protein, Adenosine deaminase, and Lactate dehydrogenase levels and pleural fluid/serum ratios in the differentiation of malignant from benign pleural effusions. İzmir Göğüs Hastanesi Dergisi, 2012;2:75-82.
  • 13- Dowers KL, Hawley JR, Brewer MM, Morris AK, Radecki SV, Lappin MR. Association of Bartonella species, feline Calicivirus, and feline herpesvirus 1 infection with gingivostomatitis in cats. J Feline Med Surg, 2010;12(4):314-321.
  • 14- Ellah MRABD, Nishimori K, Goryo M, Okada K, Yasuda J. Serum adenosine deaminase activity in bovine liver diseases, J Vet Med Sci, 2004;66(11):1421-1422.
  • 15- Farid AS, Kazuyuki Honkawa K, Fath EM, Nonaka N, Horii Y. Serum paraoxonase-1 as biomarker for improved diagnosis of fatty liver in dairy cows. BMC Vet Res, 2013;9:1-11.
  • 16- Flinn AM, Gennery A. Adenosine deaminase deficiency: a review. Orphanet J Rare Dis 2018;13:1-7.
  • 17- Foley J, Hurley K, Pesavento PA, Poland A, Pedersen NC. Virulent systemic feline Calicivirus infection: local cytokine modulation and contribution of viral mutants. J Feline Med Surg 2006;8:55-61.
  • 18- Gökce Hİ ve Bozukluhan K. Çiftlik Hayvanlarında Önemli Akut Faz Proteinleri ve Bunların Veteriner Hekimlikte Alanındaki kullanımı. Dicle Üniv Vet Fak Derg 2009;1(1):1-14.
  • 19- Kahraman MA, Gökce HI. Investigations of Adenosine Deaminase and C-reactive Protein in Cats with Feline Infectious Peritonitis. MAE J Health Sci Inst., 2020; 8,98-107.
  • 20- Kim JW, Yang IA, Oh AE, Rhyhoo YG, Jang YH . C reactive protein, sialic acid and adenosine deaminase levels in serum and pleural fluid from patients with pleural effusion. The Korean J Int Med 1988;3:122-127.
  • 21- Kodama J, Miyagi Y, Seki N, Tokumo K, Yoshinouchi M, Kobashi Y et al. Serum C-reactive protein as a prognostic factor in patients with epithelial ovarian cancer. European Journal of . Obstetrics&Gynecology 1999;82:107.
  • 22- Kulka M. A review of paraoxonase 1 properties and diagnostic applications. Polish J Vet Sci 2016;19:225–232.
  • 23- Litvinov D, Mahini H, Garelnabi M. Antioxidant and Anti-Inflammatory Role of Paraoxonase 1: Implication in Arteriosclerosis Diseases. North Am J Med&Sci 2012;4(11):523–532.
  • 24- Meazzi S, Ferriani R, Paltrinieri S, Giordano A Preliminary data about Paraoxonase-1 (PON-1) as a maker for Feline Infectious Peritonitis (FIP). Int J Health, Anim Sci & Food Safety, 2018;5(1):1-5.
  • 25- Novak F, Vavrova L, Kodydkova J, Novak F Sr, Hynkova M, Zak A et al. Decreased paraoxonase activity in critically ill patients with sepsis. Clin Exp Med 2010;10:21–25.
  • 26- Poulet H, Jas D, Lemeter C, Coupier C, Brunet S. Efficacy of a bivalent inactivated non-adjuvanted feline calicivirus vaccine: Relation between in vitro cross-neutralization and heterologous protection in vivo. Vaccine 2008;26:3647–3654.
  • 27- Radford AD, Addie D, Belák S, Boucraut-Baralon C, Egberink H, Frymus T et al. Feline Calicivirus infection. ABCD guidelines on prevention and management. J Feline Med Surg 2009;11(7):556-64.
  • 28- Reynolds BS, Poulet H, Pingret JL, Jas D, Brunet S, Lemeter C et al. A nosocomial outbreak of feline Calicivirus associated virulent systemic disease in France. J Feline Med Surg 2009;11(8):633-644.
  • 29- Rodrigues DSLF, Oliveira, MEF, Teixeira PPM, Cavalcante IJM, Vale MR). Adenosine deaminase activity as a biochemical marker of inflammatory response in goats infected by caprine arthritis–encephalitis virus. Small Rum Res 2012;108(1–3):120-126.
  • 30- Rossi G, Giordano A, Pezzia F, Kjelgaard-Hansen M, Paltrin S. Serum paraoxonase 1 activity in dogs: Preanalytical and analytical factors and correlation with C-reactive protein and alpha-2-globulin. Vet Clin Pathol, 2013;42:329-341.
  • 31- Ruiz-Gonzalez A, Bielsa S, Falguera M, Porcel JM. The Diagnostic Value of Serum C-Reactive Protein for Identifying Pneumonia in Hospitalized Patients with Acute Respiratory Symptoms. J Biomarkers, 2016;2016:1-5.
  • 32- Scavone D, Sgorbini M, Borges AS, Oliveira-Filho JP, Vitale V, Paltrinieri S. Serial measurements of Paraoxonase-1 (PON-1) activity in horses with experimentally induced endotoxemia. BMC Vet Res 2020;16:1-7.
  • 33- Schulz C, Hartmann K, Mueller RS, Helps C, Schulz BS. Sampling sites for detection of feline herpesvirus-1, feline Calicivirus and Chlamydia felis in cats with feline upper respiratory tract disease, J Feline Med Surg 2015;17(12):1012-9.
  • 34- Shunmoogam N, Naidoo P, Chilton R. Paraoxonase (PON)-1: a brief overview on genetics, structure, polymorphisms and clinical relevance. Vascular Health and Risk Management, 2018;14:137–143.
  • 35- Sproston NR, Ashworth JJ. Role of C-Reactive Protein at Sites of inflammation and infection. Frontiers in Imm
Toplam 35 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Sağlık Kurumları Yönetimi
Bölüm Araştırma Makaleleri
Yazarlar

Seda Sarıkaya 0000-0003-2128-6688

Halil İbrahim Gökçe 0000-0002-4458-0671

Proje Numarası 0577-YL-19
Yayımlanma Tarihi 31 Aralık 2021
Gönderilme Tarihi 14 Temmuz 2021
Yayımlandığı Sayı Yıl 2021 Cilt: 6 Sayı: 3

Kaynak Göster

APA Sarıkaya, S., & Gökçe, H. İ. (2021). Calicivirus ile enfekte kedilerde adenozin deaminaz 1, paraoksonaz 1, C-reaktif protein ve serum amiloid A düzeylerinin araştırılması. Veterinary Journal of Mehmet Akif Ersoy University, 6(3), 133-138. https://doi.org/10.24880/maeuvfd.971635