A New Method of Assessing Sheep Red Blood Cell Types from Their Morphology

SIMPLE SUMMARY: Haematological tests are an important diagnostic tool for animal diseases. However, little is known beyond the standardly used haematological methods in sheep, especially those in which the detection of sheep red blood cells (RBCs) shape changes is crucial. Our goal is to obtain sheep RBC morphometric parameters as well as RBC subpopulations based on their morphometric parameters. Morphometric parameters of RBC size and shape were determined from stained blood smears using SFORM, a computer-assisted program for automated cell morphometric measurements. Based on their morphometric parameters, three RBC subpopulations were obtained using principal component and cluster analysis: the smallest and most elongated RBCs, the biggest and most rounded ones, and RBCs of average size and shape. When the values of RBC haematological parameters were higher or above the physiological range, a significantly higher proportion of both average size and shape RBCs, as well as the biggest and most rounded RBCs, was obtained. Since they were obtained from healthy animals, these results indicated the importance of determining morphometric parameters of RBCs and the proportion of each RBC subpopulation, which could serve as a basis for future possibilities in the diagnostic interpretation of haematological disorders in sheep, especially those for which the detection of shape changes in ovine RBCs is crucial. ABSTRACT: Data concerning the morphometric parameters of sheep red blood cells (RBCs) obtained using computer-assisted image analysis have not yet been investigated, and there are no data on any analyses of ovine RBC subpopulations based on their morphometric parameters. The aims of this study are to determine the values of RBC haematological and morphometric size and shape parameters, to form groups according to the obtained values of haematological parameters; to determine the differences in RBC morphometric parameters between the formed groups, and to determine RBC subpopulations and their respective proportions in the formed groups. Thirty-six blood samples were collected from the jugular vein of clinically healthy Lika pramenka sheep, aged between 2 and 5 years. Haematological parameters including haemoglobin (HGB), haematocrit (HTC), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), and RBC distribution width were analysed using a haematology analyser. Haematological parameters were categorized into two groups: those with lower values or values below the physiological range (Groups 1) and groups with higher values or values above the physiological range (Groups 2). Morphometric parameters of RBCs were determined from stained blood smears using SFORM, a computer-assisted program. Significantly higher values of RBC area, outline, convex, minimal and maximal radius, as well as length and breadth were established in Groups 2 compared to Groups 1 of HGB, HCT, MCV, MCH, and MCHC, respectively. Based on the morphometric parameters of RBCs, three RBC subpopulations were obtained using principal component and cluster analysis: ES 1—the smallest and most elongated RBCs, ES 2—the biggest and most rounded RBCs, and ES 3—average size and shape RBCs. Significantly higher proportions of ES 2 and ES 3 subpopulations, as well as a significantly lower proportion of ES 1 subpopulation, were established in Groups 2 compared to Groups 1 of HGB, HTC, MCV, and MCH, respectively. It can be concluded that ovine RBC subpopulations, based on their morphometric parameters, can be obtained by using computer-assisted image analysis of RBC morphometry and multivariate statistical methods, including principal component and cluster analysis. RBC morphometry, including classification into subpopulations, could serve as a basis for future possibilities in the diagnostic interpretation of anaemic syndromes in veterinary medicine, especially in normocytic, macrocytic, and microcytic anaemias in sheep.