Exam 1- Study Guide

 Trying my best to write down the things that were being said/mentioned as well as the answers to those questions. 

*Please note to skip the first 30 seconds since it was me babbling about a video* 

Module 1

Lab safety 

•  PPE=Personal protective equipment (like lead gowns and face shields) 

1. OSHA guidelines require wherever you work to provide free PPE and safety equipment 
2. Practice chemical hygine policies should include instructions on how to use PPE 

• Know where safety data sheet (SDS) is

1. SDS has safety info that may or may not be indicated on the chemical product label, it also gives precautions, instructions and advice for handling chemical spills

• "Right to know" law---> requires you are informed of all chemicals you may be exposed to in your workplace and require PPE to be worn. PPE not optional.
• OSHA form 300a-summary of work related injuries or illnesses that happened in practice

1. anytime you get injured the incident report goes in there

• Spill clean up 

1. Keep people & pets away 
2. Do in a well ventilated area or increase it
3. DON'T use electrical equipment 
4. Wear PPE
5. Cover spill w/absorbent material & sweep into dust pan and dispose 
6. Clean depending on what SDS tells u and throw away ur PPE 
7. Replace 

• Secondary container label

1. Use when moving a chemical from its original bottle to another container 

• Medical waste vs regular waste 

1. Medical waste is waste that has stuff on it that can pose a threat to human health (ex. zoonotics, human pathogens, scalpels and needles) 

Module 2

Veterinary lab, hematology, hematic oasis 

• Sediment vs supernatant

1. Sediment= heavy stuff on the bottom 
2. Supernatant= liquid stuff on top 

• Centrifuge uses and types

1. Microhematocrit-holds capillary tubes
2.  Swing arm centrifuge-tubes swing out when in motion (Major con= gets hot fast and can damage specimen also the supernatant and the sediment usually mix when tubes go vertical

• Angled centrifuge 

○ ~52° angle 

○ Faster than swing arm, only has 1 tube size (has adapter for smaller ones) 

• What happens if you go to fast, slow, long or quick with centrifuging? 

○ Too long or fast= ruptured cells or morphology change 

○ Too slow or short= incomplete separation

• Refractometer

○ Measures refractive index of a solution 

○ Hold up to light to see ur numbers

○ Used for urine specific gravity or protein in serum

○ Calibrated by messing with thingie to make it zero 

○ Measured via refraction which is the bending of light rays as they pass through the liquid 

• Calculation 4 total magnification for objective and ocular lens 

○ Ocular magnification X objective magnification power= total magnification

• Formula for dilution 

○ To make a weaker solution from a strong one → V₁*C₁=V₂*C₂

○ Dilution factors are used to correct for having too concentrated of a sample (too much stuff for the machine to read) 

Module 3 

Erythropoiesis, Granulopoiesis, normal RBC and WBC

• Which WBC are granulocytes vs agranulocytes (granulocytes have granules in cytoplasm) 

Granulocytes Agranulocytes

Neutrophils Lymphocytes

Eosinophils Monocytes

Basophils

• Components of plasma 

○ 91% water and 9% solids

• Components of Erythrogram (RBC counts [cbc] and hematocrit which is RBC mass) 

○ CBC=Complete blood count

§ Evaluates components of the blood

○ RBC count= measures amount of RBC in blood

○ Hematocrit (Hct)= measures RBC mass as a percentage of blood 

○ Hemoglobin concentration= amount of RBC protein that carry oxygen from the lungs to the tissue in the blood

○ RBC indices: many values that describe size or erythrocytes & concentration of hemoglobin within erythrocytes (MCV, MCH, MCHC) 

§ MCV= V for volume, tells u about size of RBC

§ MCH & MCHC= tells you about hemoglobin or color of cells in ur sample

○ Platelet count: what do you think. 

• Erythropoiesis= RBC production 

• Granulopoiesis= WBC stuff. Neutrophil, eosinophil, and basophil production [granulocytes]

• Lymphocytopoiesis= Lymphocyte production 

• Megakaryopoiesis= leads to production of platelets

○ Megakaryocytes are precursors to platelets

• Active bone marrow in adult animals is at the ends of long bones & in flat bones

○ Erythropoiesis is in the liver of the embryo then spleen thymus and lymph nodes & eventually shifts to the bone marrow then granulopoiesis starts as an adult

• Pluripotent stem cell= basic undifferentiated cell in bone marrow

• Extramedullary vs medullary hematopoiesis 

○ Extramedullary= Spleen & other spots like liver and lymph node RBC production

○ Medullary= bone marrow RBC production

§ Hematopoiesis= formation of RBC

• Gen rules followed by maturing cells

○ Cells get smaller as they mature 

○ Nucleus becomes smaller and cytoplasm amount gets larger 

○ Immature cells the chromatin strands are delicate and fine (as they mature the chromatin becomes coarse) 

○ Immature cells there are nuclei found within the nucleoli and may be different sizes and numbers of em (usually round or oval & stain light blue) 

○ As the cell matures the nucleoli dissapear 

○ Perinuclear clear zone can sometimes B seen in immature cell cytoplasm 

§ Golgi apparatus makes this

• Prefixes and suffixes of cells as they mature

○ Blasts (as in myeloblasts or rubriblasts)→ Pro (as in promyelocyte)→ Cyte→ Meta→Poly (if applicable) [youngest to oldest]

• Howell Jolly Bodies

○ Small piece of leftover nucleus when its extruded 

○ Usually only found in basophilic erythrocytes (younger ones) 

• RNA found in a reticulocyte using methylene blue stain

○ As blood cells mature they lose all their DNA and RNA

• Phase right before maturity with RBC can be called a reticulocyte if RNA shows up because of the stain or a polychromatiphil or a basophilic erythrocyte

• Basophilic erythrocyte, polychromatophil and reticulocyte are all the same things

○ Reticulocyte called this when it's stained w/new methylene blue

• Erythropoietin= hormone that's made by the kidney and acts on bone marrow to stimulate erythropoiesis

• Erythropoiesis takes ~4 days

• Mature erythrocytes do not have DNA or RNA 

• Describe the appearance of the following: 

○ Segmented neutrophil

§ Has segmented nucleus w/faint blue-pink cytoplasm 

○ Band neutrophil

§ Horseshoe shaped nucleus (bands r usually neutrophilic bands) 

○ Eosinophils

§ Present in low numbers, red-reddish orange granules, cytoplasm faint blue

○ Basophils 

§ Rare. Small dark blue granules, light purple cytoplasm. Can't really see nucleus due to dark blue stuff. 

○ Lymphocytes 

§ Round, coarse clumped nucleus, super small amount of crescent shaped cytoplasm 

○ Monocytes

§ Gray-blue cytoplasm can have vacuoles in cytoplasm 

§ Largest in peripheral blood

• RBC definitions 

○ Rouleaux: Neat linear stacks of RBC (coins!) 

§ Normal in horses but not seen much in dogs and cats, and not usually seen in rats/mice, llamas and cows

○ Anisocytosis: Variation of RBC size 

§ Normal in cats, cows, rats/mice, rabbits, ferrets & guinea pigs

• Birds + reptiles have nucleated RBC (fuckin weirdos.) 

• Largest cell in bone marrow is megakaryocyte

•  Platelets or thrombocytes are fragments of the cytoplasm of megakaryocyte and have no nucleus but have granules

Module 4

WBC functions & features

• RBC function in bringing oxygen places and WBC have functions related to immune system stuff

• Where and how do granulocytes travel in the vascular system? 

○ Can move back and forth between their circulating pool and marginal pool

§ Marginal pool= stuck to the walls of the blood vessels (like plaque) 

§ Circulating pool= in the blood, this is what we see on blood smear

○ Perform their function in tissues and they can't return to the blood once they go into the tissues

○ There's a storage pool in the bone marrow waiting to be used

• Neutrophils are the most common WBC in all common domestic species besides ruminants, rats & mice

○ Lymphocytes are the most common in rats, mice and ruminants

• Lymphocytes are mostly made in lymphoid tissues from the spleen, thymus, tonsils, peyers patches and lymph nodes

○ Some are in the bone marrow 

• Lymphocytes are the 2nd most common type cell found in the peripheral blood in most domestic species

• Neutrophils → first responders

○ They phagocytize (eat), engulf and destroy things) 

○ Also think of pus & pus forming bacteria (yucky!) 

• T and B lymphocytes 

○ T lymphocytes: come from the thymus & are responsible for cell mediated immunity 

§ Destroy antigen invaders by coming into contact with them and destroying them, they're cytotoxic

○ B lymphocytes: usually come from bone marrow & are responsible for humeral immunity 

§ Make antibodies that are used 2 destroy invaders

• Macrophage= cell derived from a monocyte & functions in tissues by phagocytizing (eating) dead and damaged cells, cellular debris and large particles like fungi and protozoa

○ Like a maggot

• Leukogram includes 

○ Total WBC 

○ Leukocyte differential 

○ Absolute leukocyte counts

• Look @ blood smear to find leukocyte abnormalities

• Leukogram tells us bout the total number of WBC and each type of em and how many of em there are relative to each other 

• Calculations 

○ 90 neutrophils from 100 cells, relative and absolute differential values important 

○ Multiply percent of neutrophils by the total WBC count u get absolute number of that kinda cell

- Basically know how to use your differential to figure out and get ur total WBC count & figure out ur absolute number from that (math :p) 

Module 5

Leukocyte responses

• Left shift: increase above normal in the number of immature neutrophils in peripheral blood 

○ Note that left and right shift refers to only neutrophils

• Compensated vs Non-compensated left shift 

○ Compensated/regenerative left shift: total WBC count higher than normal → leukocytosis. Increased neutrophils w/number of mature greater than number of immature. 

§ Bone marrow keeping up w/neutrophil demands

○ Non-compensated/Degenerative left shift: total WBC count usually normal or below normal

§ Types: Neutrophilia (More immature than mature), Normal neutrophil count with immature present, neutropenia with immature present

• Look at CBC and be able to tell if u have absolute or relative increase or decrease in certain types of WBC

• Leukogram due to things 

○ Leukogram due to inflammation=inflammatory leukogram 

§ Left shift hallmark of that 

○ Leukogram due to effects of steroids: stress leukogram 

§ Can be due to stress in the body or because corticosteroids (steroids) are being given as medication

• Toxic neutrophils

○ Have increased blueness to cytoplasm, basophilia, vacuoles in cytoplasm and sometimes Dole bodies

§ Seen w/ systemic inflammation that body can't cope w/. Usually see with overwhelming infections. 

• Sequestration of neutrophils: when they all go from circulating pool to marginal pool they sequester themselves along sides of blood vessels 

○ Happens w/anaphalaxis and toxic shock 

○ Things that can influence leukocyte results: age, species, degree of excitement and stage of pregnancy 

○ Young animals have higher WBC counts 

○ Excitement &/or struggling can cause increase in total WBC count that’s caused by an increase of neutrophils 

○ WBC can be increased in late pregnancy 

• Causes of…

○ Neutrophilia: excitement, stress & inflammation

○ Neutropenia: excessive use, decreased production and sequestration 

○ Eosinophilia: parasites & allergies 

○ Eosinopenia: stress & steroids

○ Lymphopenia: stress & steroids

○ Lymphocytosis: cancer, adrenal gland disease, not having enough steroids or illness causing prolonged systemic use 

• Physiologic things that cause changes in lymphocytes 

○ Young age, fear, excitement in cats can cause a lymphocytosis as well

• Forms of lymphocytes 

○ Reactive/atypical lymphocytes 

§ Larger than a neutrophil and can be indented or cleft. Has dark blue cytoplasm & sometimes perinuclear clear zone

□ Reactive=when they are reacting to something 

□ Atypical=can be cancerous

○ Lymphoblasts 

§ Largest of em all. Have lots of nuclei

○ Small lymphocytes

§ Norm small lymphocytes have a lot of nucleus that’s super coarse & clump with crescent of cytoplasm on the side

Module 6 

Abnormalities of RBC & WBC

• Spleen is where the body takes old & damaged RBC 

• Exercise, excitement and fear in most species increase total RBC count, PCV and total hemoglobin

○ Spleen contracts & pushes out a buncha cells 

§ Most marked in horse b/c of amount of RBC reserve in they spleen

• Changes in RBC definitions 

○ Anisocytosis: Variation of RBC size

○ Poikilocytosis: Variation of RBC shape 

○ Polychromasia: variation of color

○ Hypochromasia: decrease in color (usually w/iron deficiency) 

○ Rouleaux: Neat linear stacks of RBC

§ Due to electrical charges on the RBC surface can indicate RBC didn't form right in dogs, cats and pigs

○ Agglunation: increased 3D clustering of RBC due to antigen and antibody interactions on surface of the cell

§ Can see macro (w/ur eyes) and microscopically 

§ Happens w/immune system disorder where ur system starts attacking ur RBC

○ Heinz Bodies: sometimes see w/Tylenol poisoning in cats 

○ Nucleated RBC: seen w/anemia or due to lead poisoning 

○ Spherocytes: RBC that don't have central pallor

§ Immune mediated hemolysis will cause portions of the RBC being taken away and shrink the size of the RBC down

○ Howell Jolly Bodies: Small piece of leftover nucleus when its extruded 

○ Basophilic stippling: blue staining basophilic granules in RBC

§ Can b due to lead poisoning

• MCV: indicated size of RBC (volume?) 

○ If small (lower than norm) its microcytic

○ If normal it's normalcytic 

○ If big (higher than norm) its macrocytic 

• MCHC: indicates color 

○ Hypochromic if low 

○ Normochromic if normal 

Module 7

Anemia

• Shock happens when you lose 60-70% of blood vol 

• Causes of anemia

○ Too little of something in the body 

○ Increase loss of that thing, increase destruction or use & decreased production 

• Hemolytic anemia: destruction of RBC

○ Break apart and release parts of hemoglobin

• Aplastic anemia: Increased production of RBC

○ Aplasia means not weak

○ Won't see nRBC, basophilic erythrocytes, reticulocytes

• RBC destruction= hemolysis

○ When u see reticulocytes and nRBC think chronic blood loss b/c acute blood loss there's no time for bone marrow to push out the new RBC

• nRBC and reticulocytes means bone marrow is responding

○ Regenerative anemia

• Causes of hemolysis

○ Parasites- chew apart ur RBC

○ Toxins can break em up 

○ Metabolic disorders can cause RBC to be broken up 

○ Immune mediated disorders-ur system is attacking RBC

○ Some infections

• We count reticulocytes in anemic patient b/c good indicator of effectiveness or regeneration of bone marrow activity

○ Estimate reticulocytes by looking at 5 oil immersion fields, count 30 of em (gonna be real honest I have no idea what she went into here) 

○ Regenerative anemia is over 1%

• You need to correct for anemia 

Module 8

Hemostasis 

• Hemostasis= blood clotting

• Primary hemostasis: Platelets adhere 2 whatever needs to be stopped bleeding

○ There's platelet adhesion, activation, and aggregation that ends up as a temporary hemostatic plug (clot?)

• Secondary hemostasis: Ends with the formation of fibrin to stabilize plug

○ Happens via coagulation cascade which is the work of a bunch of coagulation factors

• Extrinsic pathway: requires tissue factor 

○ Not all factors in coagulation cascade found in blood, they aint all coagulation factors there r tissue ones 

• Vitamin K dependent coagulation factors are 2,7,9,10 

• Spontaneous bleeding occurs usually below a platelet count of 20,000-30,000

○ Patient comes in sneezing blood or has blood in pee that could b due to infection or due to blood vessels in those areas are bleeding so we check their platelets to make sure they don't have an issue 

•  Disorders of primary hemostasis

○ Thrombocytopenia

○ thrombopathia (when platlets don’t work right, this is rare),

○ Von Willebrand's disease (lack of Von Willebrand's factor which important 4 hemostasis) 

• Disorders of secondary hemostasis

○ Hemophilias (inherited deficiencies of certain coagulation factors) 

○ Rodenticide toxicities (rat poison!!) 

§ Knocks out vitamin K shit

• Disorders of anticoagulation (that leads to thrombosis or DIC) 

○ DIC: Disseminated Intravascular Coagulation

○ Thrombosis: Combination of platelet fibrin & cellular debris that form within the intravascular space, happens due 2 hyper coagulability, blood stasis & endothelial injury

• Petichiation: lil purple spots that happen on the mucous memebranes that indicate disorders of primary hemostasis 

• Thromboembolism: obstruction of blood vessel by a clot

• Thrombosis: Combination of platelet fibrin & cellular debris that form within the intravascular space, happens due 2 hyper coagulability, blood stasis & endothelial injury

• Hemarthrosis: Bleeding into a joint

• Ecchymosis: discoloration of the skin due 2 bleeding or bruising underneith it

• Epistasis: nose bleed 

• Tests of primary vs secondary hemostasis

○ Primary

§ Primary platelet counts

§ Buccal mucosal bleeding time (BMBT) 

§ Plasma vWF (Von Willibrands factor) 

○ Secondary 

§ Activated partial thromboplastin time (aPTT)

§ Prothrombin time (PT) 

§ Protein induced by vitamin K antagonist (PIVKA) 

• Buccal mucosal bleeding time (BMBT) 

○ Evaluates primary hemostasis & measures how long it takes for inside of upper lip to stop bleeding

• ACT 

○ Needs blood tube that has diatomaceous earth (grey top tube) 

○ Evaluates extrinsic & common pathways of coagulation cascade

• PT/aPTT

○ Requires blue top tube with citrated plasma in them

○ PT: Evaluates extrinsic and common pathways of coagulation cascade 

○ aPTT: Evaluates intrinsic pathways of these [fibrins?]

• Dissolution of fibrin clot by plasmin as healing occurs

○ Elevated d-dimer levels mean active clot degredation

• Anticoagulation-rodentacides can inhibit vit K and affect all of the pathways (intrinsic, extrinsic and common!) 

• Estimate of platelets in blood smear

• Look at avg number of platelets in 5-10 oil immersion fields

○ 8-15 per field is normal if there's no clumping