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Hematology. Fish Health Management . Cold Blooded Vertebrates. Primary site of hematopoiesis is anterior fourth of the kidney Spleen is a accessory area of hematopoiesis as well as the site for blood cell destruction
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Hematology Fish Health Management
Cold Blooded Vertebrates • Primary site of hematopoiesis is anterior fourth of the kidney • Spleen is a accessory area of hematopoiesis as well as the site for blood cell destruction • Bone marrow, spleen and lymph nodes serve this function warm blooded vertebrates • Various types of blood cells in head kidney:- Immmature- Mature- Red Blood Cells and White Blood Cells
Blood Cell Types • Hemocytoblast- Stem Cell or blast cells,- Most abundant in hemopoetic tissue of kidney, rare in blood • Erythrocytes- Red Blood Cells- Carries Oxygen • Leukocytes- White Blood Cells- Blood Clotting- Phagocytosis- Inflammation Response
Erythrocytes • Primary Function: Oxygen transport • Elongated elliptical cell: 13-16 m long by 7-10 wide m • Nucleus centrally located, elliptical in shape • Polychromatins- Immature erythrocytes-less elliptical with blueish-gray cytoplasm
Ghost Cells • Artifact of Sampling
Leukocytes • Immune Response • Coagulation • Hard to differentiate
Lymphocytes • Usually sparse in red blood cells • Spherical in shape, 7-10 m • Eccentrically located spherical nucleus- stains reddish-purple
Neutrophils (Polymorphonuclear Leukocytes) • Only granulocytic leukocyte present in rainbow trout • Origin and function of theses cells not well understood • Accounts for 1-9% of total leukocytes • Average diameter is 9-13 m • Multi-lobuated nucleus (2-5 lobes)
Thrombocytes • Occur in varying shapes and size: elongated, spiked, fusiform, and spheroid. • Elongated - 5-8 m long • Spiked – streaked cytoplasm on one end • Fusiform- streaked cytoplasm on both ends • Spheroid- dense nuclei, little or no cytoplasm
Monocytes and Macrophages • Monocytes- large 9-25 m • Indistinguishable from Macrophages once in tissue • Cytoplasm often has granules and a large nucleus • Macrophages not usually seen in blood
Kidney Imprint • Various states of hematopoiesis
Blood Sampling Non-Lethal • Mouth Catheterization • Caudal Artery • Heart Lethal • Sever Caudal Peduncle • Sever Gill
Blood Assessment • Three Indexes: • Red Blood Cell Abundance • Hemoglobin • Hematocrit - Ratio of Packed RBC’s to total blood volume • Variations in reading can be caused by species, sex, age, temperature, clotting, water quality, season
Hematocrit Readings • Low • Indicates anemia, gill damage, osmoregulation impairment • High • Indicates stress, dehydration, hemoglobin concentrations
Lab Procedure • Take blood from caudal artery • Use one tube for Blood Smear • Seal other 2 with CritoSeal at vacant end • Measure % of both • Tissue imprints
Tissue Imprints • Cut a small piece of Tissue • Using forceps, create imprints by touching tissue to slide in succession Fish 1 K L S
Stain using Diff Quick • Fix slides in Solution A (Methyl Alcohol) for 10 min • Dip slides in Solution B 25 times • Dip slides in Solution C 25 times • Wash with DDI water. Dry and examine the slides.