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1、Veterinary Clinical Pathology,兽医临床病理学,Prof. Zhaoxin Tang,College of Veterinary Medicine, South China Agricultural University, Guangzhou, China , 510642,2021/02/24,1,Veterinary Clinical Pathology兽,Preface,Veterinary Clinical Pathology:Veterinary Laboratory MedicineInclude:1 Clinical Hematology2 Clini
2、cal biochemistry3 Clinical cytology4 Clinical microbiology5 Clinical parasitology6 Clinical toxicology,2021/02/24,2,PrefaceVeterinary Clinical Pat,Preface,General Laboratory concepts Veterinarians have many choices regarding laboratory testing. Important factors include:-Need and usefulness-Practica
3、lity-Cost-effectiveness-Accuracy-Turnaround time,2021/02/24,3,PrefaceGeneral Laboratory conc,Complete Blood Count and Bone Marrow Examination:general comments and selected techniques,Complete blood countQuantitation techniquesBlood smear analysisOther determinationsBone marrow examinationBone marrow
4、 biopsy and aspirate,2021/02/24,4,Complete Blood Count and Bone,Complete blood count (CBC),CBC is a pro tests used to describe the quantity and quality of the cellular elements in blood and a few substances in plasma.CBC is a cost-effective screen the detects many abnormalities and disease condition
5、s.Bone marrow examination is used in selected instances to answer questions the more readily available CBC cannot.,2021/02/24,5,Complete blood count (CBC)CBC,Quantitation Techniques,Sample submissionMicrohemotcritHemoglobin concentrationCell countsAbsolute nucleated RBC countAutomated hematology cel
6、l counters,2021/02/24,6,Quantitation TechniquesSample,Blood Smear Analysis,Making the smearStainsEvaluating blood smears -platelet morphology -leukocyte morphology -leukocyte estimation -leukocyte differential count -erythrocyte morphology,2021/02/24,7,Blood Smear AnalysisMaking the,Bone Marrow Exam
7、ination,Bone marrow is usually examined to answer certain question that arose from evaluating the CBC.Indications for bone marrow examination include: -nonregenerative anemia -Persistent neutropenia -Persistent thrombocytopenia -Unexplained polycythemia or thrombocytosis -Atypical cells in blood,202
8、1/02/24,8,Bone Marrow ExaminationBone ma,Erythrocytes,Basic concepts of erythrocyte function,metabolism, production and breakdownHeme synthesisGlobin synthesisIron metabolism,2021/02/24,9,ErythrocytesBasic concepts of,Erythrocyte metabolism,Embden-meyerhof pathway -Glycolysis generates ATP and NADHP
9、entose phosphate pathway -This pathway produces NADPHMethemoblobin reductase pathway -Methemoglobin(Fe3+) cannot transport oxygenRapoport-luebering pathway -2,3 diphosphoglycerate(2,3 DPG),2021/02/24,10,Erythrocyte metabolismEmbden-m,Red blood cells The fundamental stimulus for production of red blo
10、od cells (erythropoiesis) is erythropoietin(红细胞生成素）, a glycoprotein produced by the kidneys in response to renal tissue hypoxia. Other hormones, such as corticosteroids, thyroid hormone and androgens, stimulate the production or release of erythropoietin but have no intrinsic erythropoietic activity
11、.The average lifespan of a circulating erythrocyte is 110-120 days in the dog and 68 days in the cat. Aged or damaged red cells are removed primarily by macrophages in the liver, spleen and bone marrow.,2021/02/24,11,Red blood cells2021/02/2411,NeutrophilsThe production of neutrophils, eosinophils a
12、nd basophils is termed granulopoiesis. The neutrophils in the bloodstream either circulate freely (the circulating pool) or adhere to the vascular endothelium (the marginal pool). In the dog the marginal pool and the circulating pool are approximately equal in size, whilst in the cat the marginal po
13、ol is two to three times larger than the circulating pool. There is a continual exchange of cells between these two pools. The half-life of circulating neutrophils is only 6-14 hours, after which time they leave the circulation and pass into the tissue pool. The circulating time is shortened during
14、acute infections as neutrophils pass to the site of infection in the tissues. The main function of the neutrophil is the phagocytosis of pyogenic bacteria.,2021/02/24,12,Neutrophils2021/02/2412,Lymphocytes Lymphoid primitive stem cells divide and differentiate into pre-B lymphocytes and pre-T lympho
15、cytes in the bone marrow. Pre-T lymphocytes mature and proliferate into T cells in the thymus. Pre-B cells proliferate in the bone marrow and migrate to peripheral lymphoid organs (spleen and lymph nodes) where further proliferation takes place. Platelets Platelets are produced from the cytoplasm of
16、 megakaryocytes Once in the circulation, platelets survive for 8-12 days. Up to 20-30% of circulating platelets can be sequestered in the spleen; the figure may be a high as 90% if there is splenomegaly. Old or damaged platelets are removed from the circulation by the spleen, liver and bone marrow.,
17、2021/02/24,13,Lymphocytes2021/02/2413,ROUTINE HAEMATOLOGY,The complete blood count is an integral part of the diagnostic investigation of any systemic disease process. It consists of two components: A quantitative examination of the cells, including : packed cell volume (PCV) total red cell count (R
18、BC) total white cell count(WBC) differential white cell count platelet count mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), total plasma protein concentration.A qualitative examination of blood smears for changes in cellular morp
19、hology.,2021/02/24,14,ROUTINE HAEMATOLOGYThe comple,Table 1 Reference values for red cell indices,ROUTINE HAEMATOLOGY,2021/02/24,15,Table 1 Reference values for,RED BLOOD CELL INDICES,MCV(fl飞升)= PCV (L/L) 1000/ total red cells ( 1012/L) MCH (pg皮克) = total haemoglobin (g/dl) 10/ total red blood cells
20、 ( 1012/L) MCHC (g/dl) = total haemoglobin (g/dl)/PCV (L/L),RBC indices are helpful in the classification of certain anemias.,ROUTINE HAEMATOLOGY,2021/02/24,16,RED BLOOD CELL INDICESMCV(fl飞升,Differential white cell countsThe differential white cell count is performed by counting 200 leucocytes in a
21、blood smear. The cells are counted along the long edge of the smear, using the battlement meander method: four high-power fields are counted in one direction, then four more in a direction at right angles to the first, and so on, following the shape of a battlement. The percentage of each type of ce
22、ll is determined. This percentage is then multiplied by the total white cell count to obtain an absolute count for each cell type.,ROUTINE HAEMATOLOGY,2021/02/24,17,Differential white cell counts,Plasma protein concentration(Reference range: 60-80 g/1 for the dog and cat) Total plasma protein (TPP)
23、and PCV should be interpreted together. Qualitative examination of a blood smearA blood smear should always be evaluated when automated cell counts are made or when in-practice instrumentation is limited to a centrifuge for PCV Preparation of a blood smear A small drop of blood is placed on one end
24、of a glass slide, using a capillary tube. A spreader slide (made by breaking off the comer of another slide, after scoring it with a glass cutter or diamond writer) is placed on to the slide holding the blood drop, in front of the drop and at an angle of 20-40.,ROUTINE HAEMATOLOGY,2021/02/24,18,Plas
25、ma protein concentrationRO,ANAEMIAAnaemia is characterized by an absolute decrease in red cell count, haemoglobin concentration and PCV. Acute haemorrhage Acute haemorrhage may be due to trauma or surgery, bleeding gastrointestinal ulcers or tumours, rupture of a vascular tumour (e.g. splenic haeman
26、giosarcoma), or a coagulopathy (e.g. warfarin toxicity). Immediately following acute haemorrhage the red cell parameters, including PCV, are normal because both red cells and plasma have been lost in proportion. Compensatory mechanisms such as splenic contraction may further offset any fall in PCV.
27、The PCV falls when blood volume is replaced by interstitial fluid and so does not indicate the full magnitude of blood loss for at least 24 hours after the onset of haemorrhage.,ROUTINE HAEMATOLOGY,2021/02/24,19,ANAEMIAROUTINE HAEMATOLOGY202, Chronic haemorrhageChronic external blood loss (e.g. chro
28、nic gastrointestinal haemorrhage, renal or bladder neoplasia) initially results in a regenerative anaemia but gradually the anaemia becomes non-regenerative as the iron stores become depleted. Young animals become iron-deficient more bone marrow is already very active producing red cells quickly tha
29、n adults following blood loss, partly because they have low iron stores and partly because their to match their growth rate and so has less capacity to increase its rate of haemopoiesis. Haemolytic anaemiasMost cases of haemolytic anaemia are immune-mediated. In the dog most cases of immune-mediated
30、 is haemolytic anaemia (IHA) are primary (idiopathic) and are termed autoimmune haemolytic anaemia (AIHA). IHA may occur in association with: drugs(e.g. potentiated sulphonamides); lymphoreticular diseases (e.g. lymphoid leukaemia); systemic lupus erythematosus; or infections (e.g. Babesia, bacteria
31、l endocarditis).,ROUTINE HAEMATOLOGY,2021/02/24,20, Chronic haemorrhageROUTINE H,DISORDERS OF WHITE CELL NUMBER,NeutrophiliaFigure 3.20 Causes of neutrophilia Physiological response (fear, excitement, exercise) Stress/corticosteroid-induced Acute inflammatory response: bacterial infection (localized
32、 or generalized), immune-mediated disease, necrosis,e.g.pancreatitis, neoplasia, especially with tumor necrosis. Chronic granulocytic leukaemia Neutrophil dysfunction Paraneoplastic syndromes,2021/02/24,21,DISORDERS OF WHITE CELL NUMBER,Neutropenia The three main causes of neutropenia are: An overwh
33、elming demand for neutrophils Reduced production of neutrophils in the bone marrow Defective neutrophil maturation in the bone marrow. An overwhelming demand for neutrophils may occur with peracute bacterial infections, especially Gram-negative sepsis and endotoxaemia. Other possible causes include
34、peritonitis, pyometra（子宫蓄脓）, aspiration pneumonia and canine parvovirus infection.,DISORDERS OF WHITE CELL NUMBER,2021/02/24,22,Neutropenia DISORDERS OF WHITE,Eosinophilia Eosinophils are distributed in the body among various pools in a similar way to neutrophils, although the bone marrow storage po
35、ol is minimal. Eosinophils circulate in the bloodstream for only a few hours before entering the tissues, where they may live for several days. Their two main functions are to kill parasites and to regulate allergic and inflammatory reactions. Eosinopenia Eosinopenia in combination with lymphopenia
36、occurs following stress, administration of corticosteroids and in spontaneous hyperadrenocorticism (Cushings syndrome). Basophilia Basophils contain inflammatory mediators such as histamine and heparin and function in a similar manner to mast cells in hypersensitivity reactions.,DISORDERS OF WHITE C
37、ELL NUMBER,2021/02/24,23,Eosinophilia DISORDERS OF WHIT,Lymphocytosis Causes of lymphocytosis 1. Physiological lymphocytosis, with concomitant neutrophilia, in response to excitement (especially cats) 2. Strong immune stimulation (e.g. in chronic infection, viraemia or immune-mediated disease) 3. Ch
38、ronic lymphocytic leukaemia 4. Hypoadrenocortiscism (lymphocytosis may be associated with an eosinophilia) 5. Increased numbers of large reactive lymphocytes may occur transiently following vaccination 6. Young animals have a higher lymphocyte count than adult animals,DISORDERS OF WHITE CELL NUMBER,
39、2021/02/24,24,Lymphocytosis DISORDERS OF WHI,LymphopeniaCauses of lymphopenia are listed.,StressGlucocorticoid therapyHyperadrenocorticismChylothorax (loss of lymphocytes into the pleural space)Lymphangiectasia (loss of lymphocytes into the gut)Acute phase of most viral infections (e.g. canine diste
40、mper, parvovirus, FeLV)Septicaemia/endotoxaemia,DISORDERS OF WHITE CELL NUMBER,2021/02/24,25,LymphopeniaStressDISORDERS OF,Reference ranges for total and differential white blood cell counts,2021/02/24,26,DogsCats percentageAbsolute va,Table 2 shows the alterations in some of parameters in various d
41、iseases.Laboratory assessmentTests to assess primary haemostasis include: Platelet count Bleeding time Clot retraction.Tests to assess secondary haemostasis include: Whole blood clotting time (WBCT) Activated clotting time (ACT) Activated partial thromboplastin time (APPT) One-stage prothrombin time
42、 (OSPT) Thrombin time (TT),DISORDERS OF WHITE CELL NUMBER,2021/02/24,27,Table 2 shows the alterations,Disseminated intravascular coagulation (DIC): This may be triggered by a wide variety of diseases, including endotoxaemia neoplasia (especially haemangiosarcoma 血管肉瘤) acute infections (e.g. infectio
43、us canine hepatitis) haemolytic anaemia pancreatitis heat stroke. The clinicopathological features of DIC are: Thrombocytopenia Increased OSPT/APTT Elevated FDPs Low fibrinogen Schistocytes in the blood film.,DISORDERS OF WHITE CELL NUMBER,2021/02/24,28,Disseminated intravascular coa,兽医临床病理学,College
44、 of Veterinary Medicine, SCAU, Guangzhou,China 510642,Veterinary Clinical Pathology,2021/02/24,29,兽医临床病理学College of Veterinary M,Clinical biochemistry,Introduction Serum proteins Total protein and albumin Globulins Indicators of renal function Urea nitrogen Creatinine Markers of hepatic disease Alan
45、ine aminotransferase Aspartate aminotransferase Alkaline phosphatase Gamma-glutamyi transferase Bilirubin Bile acids Ammonia Pancreatic disease Amylase Lipase,Electrolytes Sodium; Potassium; Chloride Magnesium; Calcium; Phosphorus Muscle enzymes Creatine kinaseAspartate aminotransferase Carbohydrate
46、 metabolism GlucoseFructosamine Lipid metabolism CholesterolTriglycerides Miscellaneous tests Iron Lead ZincCopper Chemical profiles and test selection,2021/02/24,30,Clinical biochemistryIntroduct,SERUM PROTEINS,Total protein and albumin Physiology The circulating proteins are synthesized predominan
47、tly in the liver, although plasma cells also contribute to their production. Quantitatively the single most important protein is albumin (35-50% of the total serum protein concentration). The other proteins are collectively known as globulins. The functions of proteins are many and varied but includ
48、e maintenance of plasma osmotic pressure, transport of substances around the body (e.g. ferritin铁蛋白, ceruloplasmin血浆铜蓝蛋白), humoral immunity, buffering and enzyme regulation. Indications for assay The measurement of proteins is generally included in an initial health screen in all patients but especi
49、ally where intestinal, renal or hepatic disease or haemorrhage is suspected. Analysis Protein concentrations can be estimated in serum, plasma, urine or body fluids with a refractometer or by spectrophotometry. Serum albumin levels are measured by bromocresol green dye溴甲酚绿 binding and the serum glob
50、ulin is calculated by subtraction of the albumin concentration from the total protein concentration.,2021/02/24,31,SERUM PROTEINSTotal protein an,Reference ranges Neonates and very young animals have lower concentrations of albumin and globulins (due to minimal quantities of immunoglobulins). As the
