American Journal of Innovative Research and Applied Sciences. ISSN 2429-5396 I www.american-jiras.com
ORIGINAL ARTICLE
| Çiğdem Gül *| Murat Tosunoğlu | Hazal Bilgin |and | Bengi Baycan |
Çanakkale Onsekiz Mart University | Faculty of Arts and Sciences | Department of Biology | Canakkale | Turkey |
| Received | 11 October 2018 | | Accepted 21 November 2018 | | Published 26 November 2018 | | ID Article | Çiğdem-ManuscriptRef.1-ajira101118 |
ABSTRACT
Background: There are hematological studies on the size and number of blood cells on the Natrix tessellata species. However, in the Natrix tessellata species, detailed hematological studies, including plasma biochemistry and effect of body size on blood parameters, were not found. Objectives: In this study, changes in hematological parameters of Natrix tessellata (Tessellated water snake), which is distributed in Çanakkale, were investigated. Total body length of all samples were measured. Hemoglobin and hematocrit values, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, morphology of blood cells, glucose, calcium, cholesterol, triglyceride, creatinine, uric acid, total protein, iron, albumin, phosphorus, magnesium parameters were determined. Methods: Field trials were carried out between May and July around Karabiga. Total of 24 samples of Natrix tessellata were captured and morphologically taken from their natural environment in order to be able to perform morphological and hematological results. For evaluation of hematological parameters, approximately 1 ml of blood was taken with Caudal Ven 5 ml diameter injector, transferred in biochemistry tubes with lithium heparin, and centrifuged to separate the plasma part. Plasma biochemical parameters were determined by autoanalyzer. Erythrocyte and leukocyte counts using a hemacytometer, and hemoglobin amount Sahli method, the hematocrit value of the plasma by measuring the proportion of blood cells, the mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration calculating the mathematical hematological results. Results: The reference ranges of hematologic parameters were determined according to the growth of Natrix tessellata, and the effect of body size on hematological parameters was determined. Mean erythrocyte hemoglobin concentration and total protein values were found to be significantly positive correlations, albumin values were found to be significantly negative correlations. It was determined that the other parameters examined did not change depending on body size. Conclusions: A study involving the effect of body size on hematological parameters in Natrix tessellata has not been found, it is expected that there will be a scientific shortage of differences in some hematological parameters and it is expected to lead the hematologic studies related to body size in other snake species.
Keywords: Natrix tessellata, Plasma biochemistry, Hematology, Morphology
1. INTRODUCTION
Determination of blood parameters in reptile species as well as in different species; may be helpful in assessing the physiological, health and environmental conditions of populations [1,2,3,4,5,6].
There are hematological studies on the size and number of blood cells, plasma biochemistry on the N. tessellata species [7,8,9,10]. However, in the N. tessellata species, detailed hematological studies, including effect of body size on clinical hematology and plasma biochemistry, were not found. There are studies on the effect of body size on blood parameters in different reptile species [11,12,13,14,15,16]. However, such studies have not been found in snake species.
The purpose of this study is; the determination of the some hematological parameters (erythrocyte count, leukocyte count, hemoglobin and hematocrit values, mean erythrocyte volume, mean erythrocyte hemoglobin, mean erythrocyte hemoglobin concentration, glucose, urea, calcium, cholesterol, triglyceride, creatinine, uric acid, total protein, iron, albumin, phosphorus, magnesium) of the N. tessellata (Tessellated water snake) and determination of whether all the parameters differ according to the body size.
2. MATERIALS AND METHODS 2. MATERIALS AND METHODS 2.1 Collecting Animal Samples:The 24 N. tessellata samples studied were captured around the Karabiga (35T526225; UTM4469255; sea level) at midday between May and July 2018. Only healthy and mature animals were studied, and they were collected generally around the water resource with the method of hand capture. All the procedures on the samples were made with the permission of Çanakkale Onsekiz Mart University, Animal Experiments Ethics Committee with the decision numbered 2017/04-16. After the samples were brought to the laboratory and the necessary blood samples were taken, the snakes were left behind in the habitats from which they were collected.
Total body lenght (TBL) were made with digital caliper. In this study, a total of 24 N. tessellata samples were classified as subadult between 35-59 cm and 60-103 cm adult according to total body lengths.Total body lenght (TBL) were made with digital caliper. In this study, a total of 24 N. tessellata samples were classified as subadult between 35-59 cm and 60-103 cm adult according to total body lengths.2.2 Hematological Analyses: For the hematological analyses, 1 mL of blood from the dorsal coccygeal vein of all samples was centrifuged in a Cence L500 centrifuge at 4.000 rpm for 10 min and the plasma part was separated from the blood cells. From the plasma obtained, glucose, urea, calcium, cholesterol, triglyceride, creatinine, uric acid, total protein, iron, albumin, phosphorus and magnesium values were measured by means of an Elimat autoanalyzer.The red blood cell counts (RBC) and white blood cell counts (WBC) were carried out using a Neubauer hemocytometer, where standard Hayem’s solution for red blood cells and Turk’s solution for white blood cells were used as a diluting solution. Hematocrit (HCT) was determined using the micro-hematocrit method [17]. The tubes were then spun in a micro-hematocrit centrifuge at 12.000 rpm for 5 min and the hematocrit (HCT) was calculated with a total blood level divided by the blood cell level. Hemoglobin concentration (Hb) was measured by the Sahli method with a Sahli’s Hemoglobinometer [17]. The mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) were calculated mathematically by taking the above-mentioned results into consideration [17]. 2.3 Statistics: The descriptive statistics were conducted using SPSS (v10.0). The Pearson correlation test is used to determination of whether all the hematological parameters differ according to the body size.
3. RESULTS 3. RESULTS A total of 24 N. tessellata samples taken from the vicinity of Karabiga were divided into 2 groups according to total body lengths (35-59 cm subadult; 60-103 cm adult) and descriptive statistics of obtained hematological parameters are given in Table 1.Table 1: Descriptive statistics of hematologic results of subadult individuals.SUBADULTADULT
ParametersNMinimumMaximumMeanSDNMinimumMaximumMeanSD
TBL (cm)1035.0057.5050.238.5651460.00103.0069.5310.498
RBC (mm3)9130000.00450000.00287777.7784079.98814160000.00440000.00260000.0088055.926
WBC (mm3)101000.003100.002320.00713.052141200.003600.001957.14827.149
Hb (g/dL)103.609.205.942.225144.209.206.611.853
HCT (%)1012.0054.0031.3012.6491411.0035.0024.357.771
MCV (µ³)9821.431846.151206.11370.66014523.811650.00976.67338.706
MCH (µµg)9153.33400.00225.5485.19014131.82380.95264.8762.831
MCHC (%)1014.2930.0019.704.3141416.5738.1828.566.758
Glucose (mg/dL)1014.89246.1097.8176.0781318.74189.0073.5549.113
Urea (mg/dL)63.009.005.702.797143.4020.009.445.507
Cholesterol (mg/dL)933.03554.40229.58169.8031433.03545.28271.74194.345
Creatinine (mg/dL)10.018.201.972.42214.132.401.05.541
Uric acid (mg/dL)97.0227.0012.526.014135.0023.0010.844.640
Albumin (g/dL)102.065.963.161.12614.015.542.521.393
Calcium (mg/dL)1012.9640.9020.778.3681411.3933.5422.295.131
Triglyceride (mg/dL)1010.82336.6058.5198.4201313.59339.4054.6487.559
Total protein (g/dL)101.453.902.34.829141.176.253.011.692
Phosphorus (mg/dL)9.014.561.861.46313.101.92.97.682
Magnesium (mg/dL)10.3210.105.752.90314.989.114.152.129
Iron (mg/dL)67.9052.2024.2519.332129.30121.7040.6834.810
TBL: Total Body Lenght; RBC: Red Blood Cell Counts; WBC: White Blood Cell Counts; Hb: Hemoglobin concentration; HCT: Hematocrit; MCV: Mean Corpuscular Volume; MCH: Mean Corpuscular Hemoglobin; MCHC: Mean Corpuscular Hemoglobin Concentration; N: Number of specimens; SD: Standard Deviation.It was determined that there was a significant correlation between MCHC value and body size (P ≤ 0.05). There was a positive correlation between the statistical results and an increase in MCHC as body size increased (r = 0.454; P = 0.026). (Table 2; Figure 1).Table 2: The table presents the pearson correlation test results for blood cell counts.Correlations
TBLRBCWBCHbHCTMCVMCHMCHC
TBLPearson Correlation1-0.0120.0740.378-0.045-0.2200.2350.454*
Sig. (2-tailed)0.9570.7300.0690.8340.3130.2810.026
N24023242424232324
*: Correlation is significant at the 0.05 level (2-tailed); **: Correlation is significant at the 0.01 level (2-tailed); TBL: Total Body Lenght; RBC: Red Blood Cell Counts; WBC: White Blood Cell Counts; Hb: Hemoglobin concentration; HCT: Hematocrit; MCV: Mean Corpuscular Volume; MCH: Mean Corpuscular Hemoglobin; MCHC: Mean Corpuscular Hemoglobin ConcentrationFigure 1 : The graph showing the positive correlation between body size and MCHC value. (TBL: Total Body Lenght; MCHC: Mean Corpuscular Hemoglobin Concentration ).There was no correlation between body size and the number of erythrocytes. leukocyte count, hemoglobin, hematocrit, mean erythrocyte volume (MCV), mean erythrocyte hemoglobin (MCH) values (P≥0,05).According to the results of plasma biochemistry; total protein and albumin values were found to be a significant correlation between body size (P≤ 0,05). It has been found that there is a negative correlation between albumin value and body size, albumin value decreases as body size increases (r= -0,572; p=0,003) (Table 3; Fig 2). The total protein value was found to be statistically positive according to the results, and as the body size increased, the total protein value also increased (r= 0,424; p= 0,041) (Table 3; Fig 3). Table 3 : The table presents the pearson correlation test results in plasma biochemistry data.Correlations
TBLGlucoseUreaCholesterolCreatinineUric acidAlbuminCalciumTriglycerideTotal proteinPhosphorusMagnesiumIron
TBLPearson Correlation1-0.0720.2360.212-0.166-0.269-.572**-0.220-0.0480.420*-0.093-0.148-0.032
Sig. (2-tailed)0.7450.3450.3310.4390.2270.0030.3020.8290.041.682.4900.906
N24231823242224242324222416
**: Correlation is significant at the 0.01 level (2-tailed); *: Correlation is significant at the 0.05 level (2-tailed); TBL: Total Body Lenght.Figure 2: The graph showing negative correlation between body size and albumin value. (TBL: Total Body Lenght).Figure 3. Graph showing the positive correlation between body size and total protein value. (TBL: Total Body Lenght).There was no correlation between body size of glucose, calcium, cholesterol, triglyceride, creatinine, uric acid, iron, phosphorus and magnesium parameters (P≥0,05).4. DISCUSSION There are studies on the effect of body size on blood parameters in different reptile species. According to Bolten and Bjorndal (1992) and Fong et al., (2010), total protein, hematocrit and aspartate aminotransferase values were altered depending on body size in the species Chelonia mydas [11, 12]. Variation in aspartate aminotransferase, total protein and albumin values of Varanus varius species was observed by Scheeling et al., (2011). Hofmeyr et al. (2017) reported that the hemoglobin concentration in Psammobates geometricus was similar in female and juvenile individuals [16,13]. However, since there is no study on the effect of body size on hematological parameters in snake species, no comparison can be made with the results obtained.5. CONCLUSION A total of 22 hematological parameters were examined in the N. tessellata specimen and were determined by correlation tests in which 3 hematological parameters showed significant changes depending on the size.As a result, it was determined that the body size of N. tessellata was affected on some hematological parameters. MCHC and total protein values were found to be significantly positive correlations, albumin values were found to be significantly negative correlations. 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