111J Contemp Med Sci | Vol. 2, No. 7, Summer 2016: 111–114 Research Histological study of granulocytic series in the bone marrow of adult goat (Caprus Hircus) Yahia Dahash Hamdi,a Alzubaidi KA,b Siraj M Al-Kafagyb ISSN 2413-0516 Introduction The granulocytic sequence in the active bone marrow is con- fined to the selected areas of bones, such as the flat bones of the head, pelvis, ribs, vertebral bodies, and the ends shift of the long bones. Indeed, the blood cells also produced in the spleen and liver of the fetus.1,2 In adult, the normal marrow consists of hematopoietic precursors of erythrocytes, granulocyte, some lymphocytes, plasma cells, and few scattered precursor of platelets.3,4 Generally, hematopoiesis is a complex process in which pluripotent stem cells differentiate and evolves into committed progenitor cells. They in turn proliferate into mature functional blood cells such as erythrocyte, megakary- ocytes, monocytes, neutrophils, esinophils, basophiles, T- as well as B- lymphocytes. Thus, hematopoiesis takes place within the bone marrow stromal microenvironment.5,6 Materials and Methods Five adult goats were used in this research. These animals were obtained from the Animal’s Field in the College of Veterinary Medicine, University of Al-Anbar. The adult goat weight (23–28) kg. These animals are supplied by ruminates diet supple- ment with vitamin C. The goats are maintained in a pathogen- free environment and femurs are chosen for bone marrow aspiration and biopsy. The great advantage is that serial exam- inations showing distinct qualitative differentiation of marrow cells may be obtained from femoral bone. Each goat was anes- thetized with a mixture of Xylazine hydrochloride 0.1 mg/kg B.W. and Ketamine hydrochloride 4 mg/kg B.W. intramuscu- larly.7 The hind limb was shaved and prepared aseptically with providone iodine. A small incision is made through the skin to facilitate the entering of the needle. The tip of the needle was pushed through the head of the femur until it reaches the shaft aDepartment of Anatomy, Histology and Embryology, College of Veterinary Medicine, University of Al-Fallujah, Al-Anbar, Iraq. bDepartment of Anatomy, Histology and Embryology, College of Veterinary Medicine, University of Al-Qasim Green, Babylon, Iraq. Correspondence to Yahia Dahash Hamdi (email: yahia83mwr@yahoo.com). (Submitted: 7 June 2016 – Revised version received: 10 July 2016 – Accepted: 14 July 2016 – Published online: 26 September 2016) Objectives The aim of this research is to identify and evaluate the cellular granulocytic series of bone marrow in goat that aids to diagnose bone marrow from a variety of clinical disorders in the field of medicine. Methods Five adult goats were used in this research. These animals are obtained from the Animal’s Field in the College of Veterinary Medicine, University of Al-Fallujah. Results The first indication of the granulocytic series begins from Myeloblast, which possesses eccentric, round or oval nucleus that surrounded by a rim of cytoplasm. The smaller nucleus leads to identify the Promyelocyte. The cytoplasm of Promyelocyte is very light. Definitive granulations appear within the cytoplasm of Myelocyte, which has round to oval nucleus. The elongation of the nucleus indicates the Metamyelocyte. This cell shows the neutrophilic, eosinophilic and basophilic Metamyelocyte. A large segment of a circle or twisted nucleus represent the Band cell. The last stage of granulocytic series is the segmented cell, which is illustrated by segmented neutrophil, eosinophils, and basophils. Conclusion The developmental process of granulocytopioesis series of bone marrow in goat encompasses a lineage of successive morphological alterations involves nuclear and cytoplasmic changes as well as granule transformation resulting in the production of granulocytes. Keywords developmental process, granulocytic series, bone marrow, adult goat Caprus Hircus of femur. Special syringe is applied to obtain bone marrow aspiration. Syringe is then withdrawn and the marrow expelled onto glass slides. Marrow then spreads between the slide and the resultant smear are rapidly air dried.8 All smears were stained with Giemsa stain.9 Results and Discussions The granulocytic events or myeloid series begin with the for- mation of Myeloblast (Fig. 1) which is the first precursor cells formed by uni-potent stem cell that has the capacity to give rise to all the granulocytic cell types. Myoblast is an oval cell; the cytoplasm is pale, light blue green in color and form a complete collar around the nucleus. The round or oval nucleus in which chromatin is evenly distributed and may form a velvety texture to the nucleus. The promylocyte or progranulocyte possess light cytoplasm and had a slightly smaller nucleus (Fig. 2). Small nucleoli may be present and its chromatin forms a meshwork. The Myelocyte can be recog- nized as neutrophilic, esinophilic, and basophilic Myelocyte (Fig. 3–5). The Myelocyte has a round to oval nucleus. At this stage, the neutrophilic and esinophilic Myelocytes have all the granules or very distinct, but the basophilic Myelocyte has only a few dark purple specific granules. Regardless of cell type, the granule tends to accumulate near one side of the nucleus. The Metamyelocytes are smaller cells; the nuclei tend to be elongated and show a definitive indenta- tion with the absence of nucleoli. The arrangement of the nuclear chromatin appears coarse, with definite clumping. Th e cytoplasm of Metamyelocytes shows a few granules (Fig. 6). Band cells or stab cells are characterized by neutral cytoplasm 112 J Contemp Med Sci | Vol. 2, No. 7, Summer 2016: 111–114 Histological study of granulocytic series in the bone marrow of adult goat (Caprus Hircus) Research Yahia Dahash Hamdi et al. Fig. 1 Myeloblast (arrow). ×100 Giemsa stain. Fig. 2 Promylocyte. ×100 Giemsa stain. Fig. 3 Neutrophilic Myelocyte (arrow). ×100 Giemsa stain. Fig. 4 Eosinophilic Myelocyte. ×100 Giemsa stain. Fig. 5 Basophilic Myelocyte (arrow). ×100 Giemsa stain. Fig. 6 Metamyelocytes. ×100 Giemsa stain. containing many granules and appear in the form of neutro- philic, basophilic, and eosinophilic band cells. The cyto- plasmic granules course and clumped. Their nuclei appear rod shape, kidney-bean shape or in the form of “C” shape of band cell neutrophils (Fig. 7). Band cell can be identified by their own segmented nuclei (Fig. 8). These results were in Fig. 7 Neutrophilic band cell (arrow). ×100 Giemsa stain. Yahia Dahash Hamdi et al. 113J Contemp Med Sci | Vol. 2, No. 7, Summer 2016: 111–114 Research Histological study of granulocytic series in the bone marrow of adult goat (Caprus Hircus) agreement with the previous result.10 Eosinophilia granules are found in the cytoplasm of eosinophilic band cells (Fig. 9). Their nuclei have two segments or in the form of horseshoes. This result coincides with published report.11 The segmented cell showed the form of the three granulocytic series, and it Fig. 9 Esinophilic band cell (arrow). ×100 Giemsa stain. Fig. 10 Neutrophil (arrow). ×100 Giemsa stain. Fig. 11 Eosinophil (arrow). ×100 Giemsa stain. Fig. 12 Basophil (arrow). ×100 Giemsa stain. Fig. 8 Band cell basiophilic. ×100 Giemsa stain. represented the last stage of myeloid cell maturation the neu- trophil has a segmented nucleus, possess three to five nuclear lobes joined by short chromatin filaments (Fig. 10). The multilobulated nucleus shows a densely packed chromatin. The eosinophilic segmented cells possess two nuclear lobes (Fig. 11). The nucleus of basophilic- segmented cell may appear as S-shape or consists of multiple lobes (Fig. 12). These marked results in this research were compatible with12 and with.13 Medical researcher14 registered that the granulocyte undergo many stages in the granulocytic event but they did not mention the segmented cells in their events. Conclusion The developmental process of granulocytopioesis series of bone marrow in goat encompasses lineage of successive mor- phological alterations involves nuclear and cytoplasmic changes as well as granule transformation resulting in the production of granulocytes. The earliest recognizable pro- genitor of this granulocytic series were the myeloblast. The daughter cells of myeloblasts were the promylocytes, fol- lowed by myelocytes, metamyelocytes, band cells and seg- mented cells. n References 1. Moore MC. Clinical implication of positive and negative hematopoietic stem cell regulators. Blood J. 1991;78:7–23. 2. Williams DA. In research of the self-renewing hematopoietic stem cell. Blood J. 1991;17:269–299. 3. Tavassol M, Yoffey J. Blood marrow barrier. In bone marrow structure and function, AR Liss press, New York, U.S.A. 1983;85–102. 4. Lund JE. Toxicologic effects on blood and bone marrow. In Schalms veterinary hematology, 5th ed. Lippincott press, Philadelphia, U.S.A. 2000; 44–50. 5. Morrison SJ, Hemmat HD, Wandyez AM, Weissman IL. The purification and characterization of fetal liver hematopoietic stem cells. Proc Nat Acad Sci. 1995;92:10302–10306. 6. Gasper PW. The hemapoietic system. In: Feldman BF, Zinkl JG, Jain NC, eds. Schalm’s Veterinary Hematology, 4th ed. Philadelphia. U.S.A. Lippincott, Williams & Wilkins. 2000:63–69. 7. Riebold TW. Ruminant anesthesia. In: Greene SA, ed. Veterinary Anesthesia and Pain Management Secrets. Philadelphia, PA: Hanley & Belfus. 2002; 253–262. 114 J Contemp Med Sci | Vol. 2, No. 7, Summer 2016: 111–114 Histological study of granulocytic series in the bone marrow of adult goat (Caprus Hircus) Research Yahia Dahash Hamdi et al. 8. John WH. Atlas of veterinary hematology blood and bone marrow of domestic animals. Elsevier press limited, Philadelphia, USA. 2001. 9. Galger AE, Kazolof EN. Essential of practical microtechnique. Lea and Febiger. Philadiphia, USA. 1964. 10. Borregaard N, Lollike K, Kjeldesen L. Human neutrophil granules and secretory vesicles. Eur J Heamatol. 1993;15:187–198. 11. Bruyn PP. Structural substrates of bone marrow function. Hematol. 1981;18:173–179. 12. Panopoulous AD, Watowich SS. Granulocyte colony-stimulating factor; molecular mechanisms of action during steady state and emergency hematopoiesis. Cytokines. 2008;42:277–288. 13. Karasuyama H, Kaori M, Kazushige O, Yusuk T, Yohei K, Yoshiyuki M. Roles of basophils in immediate - and delayed – onset allergic reactions. Allergy J. 2010;3:73–80. 14. Porter RL, Calui LM. Communication between bone cells and hematopoietic stem cells. Arch Biochem Biophys. 2008;473:193–200.