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Veterinary Hematology 101 Back To The Basics So You Too Can Become An Expert!

By: Lon Bartoli, AHT, BSN, VCLS, EMT-I





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Back To The Basics So You Can Become An Expert and Make Sound Clinical Decisions



Lon Bartoli, BSN, AHT, VCLS, EMT



Veterinary Hematology is more than just blood cells. Blood, highly functional and truly definitive,



does much more than provide for the transport of cellular metabolites and waste products. Blood is made up of four major components: Plasma, Red Blood Cells, White Blood Cells, and Platelets.



Each component possesses its own diagnostic significance in the veterinary clinical setting.



Without blood, you have no viable mammal life. Therefore without blood, the clinician has no valid



clinicalpicture of disease-state. To appreciate the value of blood, the clinician should examine each individual component of blood separately to gain an appreciation for the clinical significance and diagnostic value of veterinary hematology in their practice. Anemia will also be discussed.



Plasma



Plasma carries blood and blood proteins. Aside from a high water content, plasma also contains



dissolved salts, calcium, sodium, magnesium, and potassium. Plasma contains clotting factors



and on exposure to air it will clot. Serum is the clear fluid that separates from clotted whole



blood and clotted plasma. Plasma comprises approximately 20% of the animal body's extracellular fluid. Most plasma protein molecules are too large to pass through the capillary walls into the interstitial space. The small amount of protein that can pass through the capillary walls is primarily taken up by the lymph nodes and eventually returned to the circulation.



The Majority of the plasma proteins are produced in the liver. Plasma proteins form three major



chemical groups (fractions) and have varying functions.





*albumin - approximately 60%



*fibrinogen - approximately 4%



*globulins - approximatly 36% over three subfractions (IgA, IgB, & IgG)





The relative proportions of plasma proteins can vary in certain diseases and these variations can



be clinically useful in determining proper IV therapy. Albumin is the smallest of the plasma proteins and easily passes through capillary walls. In kidney disease, large amounts of albumin are excreted through damaged kidney tubules and can be detected in the urine.





Functions of the plasma proteins include :



Intravascular collid osmotic pressure. Maintains fluid and electrolyte levels.



Transport of insoluble substances allowed by protein binding processes



Contribution towards the plasma viscosity



Inflammatory response via microbe fighting antibodies



Protein storage reserve



Clotting



Protection from infection via plasma gamma globulins





Plasma also contains inorganic ions, which are important in regulating cell function and



maintaining homeostasis. As an example, depletion of potassium may occur following severe



diarrhea and vomiting. Potassium is an essential element of cell excitability. Sharp decreases in



potassium will cause muscle weakness and cardiac abnormalities. Similar problems may cause



sodium depletion. Subtherapeutic sodium levels in the plasma will result in the volume of



extracellular fluid to decrease which will lead to a drop in blood pressure causing lethargy,



dizziness, weakness and fainting.





Plasma carries a wide range of substances including dissolved gasses left over from the



respiratory exchange cycle (mostly CO2). Blood carries oxygen because it does not have an



affinity for plasma related to its water solubility.





 



Nutrients, the most abundant being glucose, are carried in the blood plasma as a source of



fuel for cellular metabolism. Amino acids, fatty acids, triglycerides, cholesterol and vitamins



are also carried by plasma. Urea, uric acid, creatinine from the kidneys, bilirubin from the gall



bladder and other waste materials are also transported by plasma. Plasma proteins carry



hormones, such as cortisol and thyroxine. The plasma also carries certain drugs and ETOH.



Platelets



Platelets are the result of cellular fragments shed from the megakaryocyte while in the bone



marrow. Platelets considered cell fragments rather than actual cells, play a critical role in blood



clotting. When an injury to the body occurs, a chemical substance is released at the site of injury.



Platelets are able to quickly adhere to this chemical and begin to form alliances with other



platelets and clotting factors. This alliance is the body's defense against bleeding to death.



Platelets are also significant in forming diagnostic clues to the blood smear and can be useful at



guiding the clinician in care planning, treatment and further diagnostic steps. Platelet morphology



together objective data can be indicative of bleeding disorders and leukemia.



RBCs



Red Blood Cells, seemingly basic, are created and have the sole purpose of keeping the mammal



alive by carrying oxygen to the tissues and white blood cells out of the bone marrow and into circulation. Red Blood Cells along with other blood components are present in nearly every portion of the body. When there is not enough blood in the body, anemia occurs and the animal begins to have clinical signs. It becomes imperative that clinicians immediately identify the etiology of anemia in order to help define or refine treatment. In doing so, the clinician will examine the blood smear and available objective data in order to quickly determine whether the anemia present as defined by a low pack cell volume (PCV) is one of production, consumption, sequestration or destruction. We will be discussing the cellular size, shape, color and other diagnostically significant data present in various states of anemia to aid the clinician in accurate slide evaluation.



WBCs



The white Blood Cell (WBC) plays an important role in the animal body by providing our bodies



with a weapon to fight against infection and disease. The primary function of the WBC is served



mostly after it leaves the marrow and enters the blood stream after being carried by the RBC from



its site of formation in the marrow, to its site of labor in the blood stream. There are five types of



white blood cells seen in blood and each has different roles to perform.



The Neutrophil



The neutrophil, in conditions of healthand certain disease, is usually the most common



granulocyte found in blood. The cytoplasm of the neutrophil contains three differing types of



granules. It is these granules that result in it being termed a granulocyte. Neutrophils generally



have segmented or hyper-segmented nuclei giving them the appearance of being mutlinucleated.



In fact, they are not multinucleated as a thin strand of chromatin connects each lobe of the



prominent dark purple, multilobed nucleus. At times, this chromatin strand can be visualized by



most microscopes, when care is taken to look for it. Sometimes however, the strand becomes



obscured by parts of the nuclei itself as a result of cell orientation and smear technique.



The three type of granules seem in the cytoplasm of the cell perform specific functions.



Primary granules are non-specific and contain lysosomal enzymes, defensins, and some



lysozyme. The granules are similar to lysosomes. They stain violet in color when prepared with



Wright's stain or Diff Quik. The enzymes produce hydrogen peroxide, which acts as a powerful



antibacterial agent.





Secondary granules, found in the cytoplasm of the neutrophil, stain neutrally a light pink. They



contain collagenase, which helps the cell to move through connective tissue, and deliver



lactoferrin, which is toxic to bacteria and fungi.



Tertiary granules have only recently been appreciated as a granular component to granulocytes.



They are thought to produce proteins, which help the neutrophil to stick to other cells and hence



aid the process of phagocytosis.



Neutrophils, once they arrive at an area of infection, respond to chemicals (called chemotaxins



which are released by bacteria and nectrotic tissue cells) and travel towards the area of highest



concentration of infection or necrotic tissue. Once they arrive at their destination, they begin the



process of phagocytosis in which the offending cells are engulfed and destroyed by powerful



enzymes. This process requires much energy, so the glycogen reserves of the neutrophil are



soon depleted and the neutrophil promptly dies soon after the phagocytotic process. When



neutrophils die, their contents spill out into the blood stream and remnants of their enzymes



cause liquefaction of closely adjacent tissue. This results in an accumulation of dead neutrophils,



tissue fluid and abnormal materials that is known as pus.



types but differ in the location in which they sequester for their function.



Helper T-Lymphocytes originate in the thymus and produce long living T-Cells which become



Killer T-Cells or K-Cells which mediate antibody dependent cell cytotoxicity (tumor rejection).



B-Lymphocytes are localized in the corticomedullary region of the lymph nodes and are made up



of cells of the germinal centers of the cortex in lymph nodes, in the red pulp of the spleen, and in

More of this tect are available at www.lonbartoli.com and www.veterinary-hemato logy.com

the submucosal regions of the stomach and respiratory tract.



Lymphocytes, distinguished by having a deeply staining purple nucleus that is sometimes



eccentrically located, usually contain a relatively small amount of cytoplasm. The small ring of



the cytoplasm contains numerous ribosomes and readily stains blue with Wright's Stain or



Diff-Quik. Small numbers of granules may also be noted in the cytoplasm randomly.



Lymhpocytes increase in number as a response to viral infection. The small lymphocyte will



approximately the same size as the normocytic RBC. The cytoplasm is often not visible because



it is obscured by the nucleus of this cell. This cell is definitively round under examination and



lacks "divets." There can be variations in the size of the lymphocyte in the k-9/f-9 with the small



type usually being the predominant type. In the small lymphocyte the chromatin is usually so



coarse that it is masked. The medium and large forms of the lymphocyte often appear smudged.



Lymphocytes will increase in number with restraint: physical or chemical, and you will usually



notice a corresponding increase in PMNs.



Anemia



Anemia is defined as a below standard hematocrit (HCT). A species specific hematocrit (1) is as



follows: Dog: 37-55, Cat: 24-45, Horse: 32-52, Porcine: 24-46, Bovine: 24-46. There are further



variations of this data available that are further differentiated on the basis of age and sex. The



author uses a combination of sources, which he has found through experience to be clinically



reliable and accurate.



Most anemic conditions (except hemorrhage anemia) can be ordered to have an etiology of



consumption, production, destruction or sequestration and further differntiation of anemic types



are considered by ascertaining variation is size, shape, color.



Anemias of consumption include the hemolytic anemias and those created by disease conditions



should as DIC and parasites where platelets and other clotting factos are consumed. Some



anemias which sequester platelets and blood to the spleen, have also been placed in this



classification, but etiologic differentiation has been found to be of clinical significance.



Anemias of destruction such as Autoimmune Mediate Hemolytic Anemia (AIMHA) exist when the



body's own antibodies destroy its own red blood cells.



Whether or not you use your own in-house clinical lab or send your specimens out to a reference



lab, this lecture will bring you back to the basics and help you remember that which you may have



forgotten in school. Many clinicians find this lecture and format helpful to expand on basic



knowledge and clinically apply what they see either under the microscope or on the lab report.



Not having adequately available time to donate towards the lab, more clinicians are relying upon



technicians to interpret laboratory results being unsatisfied with the time investment required to



await the return of results from distant reference labs. Clinics and hospitals are using this



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