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The Diagnostic Importance of Coagulation Parameters in Cattle Having Natural Theileriosis

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The Diagnostic Importance of Coagulation Parameters in Cattle Having Natural Theileriosis Polish Journal of Veterinary Sciences Vol. 20, No. 2 (2017), 369–376 DOI 10.1515/pjvs-2017-0045 Original article The diagnostic importance of coagulation parameters in cattle having natural theileriosis V. Gunes, A.C. Onmaz, I. Keles, K. Varol, G. Ekinci Erciyes University, Faculty of Veterinary Medicine, Department of Internal Medicine, Kayseri, 38100 Turkey Abstract The purpose of this study was to determine the diagnostic importance of coagulation parameters in cattle with natural theileriosis. Nine Holstein cross-breed cattle with theileriosis as infected group and 6 healthy Holstein cattle as control group were used in the present study. Mean fibrinogen level, thrombin time (TT), activated partial thromboplastin time (aPTT) and prothrombin time (PT) were not statistically different when control and infected groups compared, except for the D-dimer concen- tration. Quantitative D-dimer concentrations were determined by immune-turbidimetric assay. D-dimer values increased significantly (p<0.05) in infected group (631.55 ± 74.41 μg/L) compared to control group (370.00 ± 59.94 μg/L). D-dimer sensitivity and specificity were also determined at cut-off concentrations (372 μg/L). Sensitivity and specificity of D-dimer values were determined to be 88.89% and 83.33%, respectively. D-dimer is thought to be important indicator in the evaluation of the prognosis in theileriosis cases. Analysis of D-dimer values before and after treatment in controlled case studies were suggested in future studies to enlighten the issue. Key words: cattle, coagulopathy, D-dimer, haemolysis, theileriosis Introduction common. Haemolytic anaemia, icterus and death has been reported in the advanced stages of the disease Theileriosis is a protozoan disease transported to (Keles et al. 2009, Ozkan et al. 2013). the animals by Ixodidea ticks and caused significant Coagulant and anticoagulant systems are balanced economic losses in cattle breeding in all over the under physiological conditions (Noyan 2012, Dhanun- World (Preston 2001, Mehlhorn 2008, Orkun et al. jaya et al. 2013). When this balance is disrupted for 2012). Theileria (T) species are seen wide spreadingly, any reason, fibrinolytic system is activated. Fibrin oc- particularly in the tropical and subtropical countries curred as a result of coagulation is degraded by some (Ozkan et al. 2013, Akat et al. 2014). High fever, en- enzymes such as plasminogen and it is converted to larged superficial lymph nodes, increase in heart and fibrin degradation products (FDP) (Noyan 2012). respiratory rate are commonly seen in animals with D-dimer is one of the final breakdown products of this theileriosis. Hyperaemia or paleness in the conjunc- fibrin network. It is stabilized by factor 13. D-dimer tive tissues or mucosa and petechial haemorrhages es- contains some cross-linked components occurring pecially in mucosa and hairless regions of the skin are during stabilization. These parts are released from the Correspondence to: I. Keles, e-mail: [email protected] 370 V. Gunes et al. clot as a result of the activation of plasmin and par- Six clinically healthy Holstein cattle (Group 2: ticipated in blood flow (Ver Elst 2002, Noyan 2012). Control group) were also sampled during the same It is well known that levels of D-dimer and other period as control group. coagulation parameters increase in infection, tu- mour, after surgical operations, trauma, burn, dis- seminate intravascular coagulopathy (DIC), venous Animals thromboembolism (VTE), ischemic cardiopathy, congestive heart failure, haemolysis, bleeding, acute This study was conducted on cattle with theilerio- respiratory syndrome, liver and kidney disease, in- sis. A total of 9 Holstein cross-breed cattle from flammatory bowel disease (Noyan 2012, Dhanunjaya a herd in Kayseri province consisted of animal ma- et al. 2013). terial. Death of a few individuals and some sick ones In theileriosis, haemolytic anaemia or haemolysis was reported in the mentioned herd. In the clinical is caused by immune-mediated destruction of affected examination of nine sick animals and after micro- erythrocytes (Hooshmand-Rad 1976). Other possible scopic examination of the blood smears of the ani- mechanisms in inducing anaemia have been suggested mals, theileriosis was diagnosed. Additionally, coagu- as high osmotic fragility of erythrocytes, acceleration lation profiles and D-dimer levels of the cattle with of clearance and the presence of haemolytic activity in theileriosis were investigated in this herd. These ani- cattle highly infected with T. sergenti (Shahnawaz et mals were aged 1.5 ± 0.5 years old and comprised of al. 2001, Shiono et al. 2003). El-Deep and Younis 2 males and 7 females which weighting from 250 kg to (2009) showed a significant decrease in red blood cells 500 kg (Group 1). Six healthy Holstein cattle (2 male (RBC) and/or haemoglobin (Hb) concentrations and 4 female) aged between 2.0 ± 0.5 years old and which were determined in Egyptian buffaloes infected weighting from 250 kg to 550 kg were also used from with Theileria. In another study, Grewal et al. (2005) the different herd as a healthy control in the present showed that oxidative stress in erythrocytes of cattle study (Group 2). These animals were selected from infected with T. annulata might be the cause of in- those which body temperature, pulse and respiration creased erythrocyte fragility and membrane lysis. rate was in normal references range and which were in DIC is a syndrome characterized by stimulation of good condition. All animals with theileriosis were intravascular coagulation and degradation of haemos- treated after collecting blood specimens. For the tatic balance for various reasons (Yuksel et al. 2009). treatment; Buparvaquone (Tailerol/Provet/Turkey) Especially, it is known to occur in protozoal infections 2.5 mg/kg body weight, two doses were injected intra- such as theileriosis and babesiosis in cattle (Stockham muscularly at 48 hours intervals. Additionally for et al. 2000). Therefore, it is hypothesized that serum piroplasm form of Theileria, two doses of oxytetracyc- levels of coagulation parameters may be a useful bio- line, (Primamycine-La®, Pfizer) 20 mg/kg, BW/day in- marker and analytical parameters in the pathogenesis tramuscularly was injected in 48 hours intervals and of theileriosis cases. In this study, it was aimed to de- supportive treatment was carried out for 5 days. termine the levels of D-dimer and other coagulation The diagnosing of Theileriosis: Blood smears ob- markers in the diagnosis of DIC in naturally infected tained from blood samples of infected animals ear tips cattle with theileriosis. Furthermore, in the present stained with Giemsa staining were investigated for the study it was also aimed to investigate whether coagu- presence of piroplasm forms of Theileria spp with im- lation profile can be used as diagnostic markers or not mersion objective (x100). The pictures of smears in- in determining the presence of DIC which may deve- cluding piroplasm forms of Theileria in the eryth- lop in theileriosis. rocyte were taken by a video microscope (Zeiss Ax- ioCam ERc5s). Ten ml blood specimens were also obtained from jugular vein of Theileria-positive ani- Materials and Methods mals and healthy animals for analysis of hemacounter into the tubes with K2 EDTA 18 mg (BD The present study was conducted in the province Vacutainer). Blood specimens (3.5 ml) were also of Kayseri where theileriosis outbreak occurred dur- taken for analysis of coagulation parameters into ing May 2015 in a farm. Nine Holstein cross-breed tubes containing citrate with vacuum (Vacutest). cattle with clinical signs of theileriosis (Group 1: These tubes were blue-capped tubes containing 3.2% group with theileriosis) referred to the Clinics of In- (0.109M) sodium citrate. Plasma specimens were har- ternal Medicine, Veterinary Faculty, University of Er- vested after centrifugation of the blood specimens for ciyes, were included in this study. The disease was 15 minutes, at 4000 rev/min (Hettich ROTOFIX diagnosed based on clinical examination and labora- 32A). These specimens were stored at -20oC until the tory confirmation by Giemsa staining of blood smears. assay process and analysed after thawing immediately. The diagnostic importance of coagulation parameters... 371 Table 1. Mean haematological values in Theileria infected and healthy cattle and P values. Group 1 (n=9) Group 1 (n=6) Hematologic Parameters x¯ ± SE x¯ ± SE P values M (min-max) M (min-max) WBC (109/l) 9.18 ± 1.28 9.60 ± 1.25 9.00 (1.90-15.60) 11.40 (5.80-12.00) 0.797 Lymphocyte (%) 26.71 ± 3.54 52.43 ± 4.70 23.30 (15.30-48.10) 54.30 (37.20-67.60) 0.002 Monocyte (%) 10.07 ± 1.63 8.26 ± 0.89 10.90 (2.10-18.80) 8.20 (5.60-10.90) 0.320 Granulocyte (%) 61.14 ± 4.13 38.08 ± 4.57 62.30 (40.00-73.70) 36.05 (24.80-52.20) 0.012 RBC (106/l) 6.90 ± 0.89 7.65 ± 0.88 7.08(3.66-11.98) 7.02 (5.67-11.77) 0.689 HG (g/dl) 10.96 ± 1.31 10.63 ± 0.39 10.90 (4.80-16.80) 10.40 (9.70-12.30) 1.000 HCT (%) 34.12 ± 4.4 33.68 ± 1.83 31.00 (14.60-54.50) 33.40 (27.70-41.40) 0.529 MCV (fL) 89.58 ± 9.50 45.53 ± 3.11 47.90 (40.20-401.00) 45.55 (35.20-56.10) 0.328 MCH (pg) 15.88 ± 0.95 14.40 ± 1.10 14.40 (13.10-21.20) 14.15 (10.40-18.80) 0.529 MCHC (g/dl) 32.35 ± 0.49 31.73 ± 1.04 31.70 (30.20-35.10) 31.00 (29.40-35.70) 0.529 RDW (%) 16.51 ± 0.26 19.38 ± 0.83 16.30 (15.40-17.90) 19.60 (16.70-22.50) 0.008 PLT (109/l) 371.55 ± 52.65 456.33 ± 103.46 372.00 (70.00-563.00) 365.00 (263.00-939.00) 1.000 MPV (fL) 5.14 ± 0.12 12.81 ± 7.51 0.529 5.10 (4.50-5.90) 5.25 (4.90-6.10) PDW 16.32 ± 0.18 16.15 ± 0.22 16.30 (15.70-17.50) 16.05 (15.60-17.00) 0.607 PCT (%) 0.19 ± 0.028 0.23 ± 0.04 0.20 (0.03-0.29) 0.20 (0.13-0.46) 0.689 RBC; Red Blood Cell, HCT; hematocrit, Hgb; haemoglobin concentration, MCV; mean corpuscular volume, RDW; erythrocyte distribution change, MCH; mean corpuscular haemoglobin volume, MCHC; mean corpuscular hemoglobin concentration, WBC; White Blood Cell, PLT platelet, MPV; Mean platelet volume, PDW; platelet distribution width, PCT; plateletcrit Values are presented as means ± SE and M (min-max).
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