VLD 411.docx

CLINICAL SIGNIFICANCE &INTERPRETATION OF SERUM

1. SERUM GLUCOSE:Clinical Significance 

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Glucose determination is mainly useful in the diagnosis of diabetes mellitus in which blood glucose levels are elevated. Other diseases like hyperthyroidism, hyperpituitarism, severe neph ritis, pancreatitis, asphyxia, anesthesia, pneumonia, dehydration and certain hepatic disorders also lead to hyperglycemia. Hypoglycemia occurs frequently as a result of ov er dosage of insulin or antidiabetic treatment. Hypoglycemia may also be noticed in c onditioned like hyperinsulinemia, hypothyroidism, hypopitutarism and hypoadrenocorticism. Hypoglycemia may also occur in certain physiological conditions like pregnancy, starvation, severe prolonged exercises and lactation.

2. LIPIDS:-

Clinical Significance 

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High values of total cholesterol may be found in diabetes mellitus, hypothyroidism, obstructive jaundice, nephrotic syndrome, biliary cirrhosis, atheroslerosis, etc. Low values of total cholesterol may be found in hyperthyroidism, malnutrition, gaucher’s disease and acute hepatitis. Decreased level of HDL cholesterol lead to inc reased chance of coronary heart disease (CHD) while increased levels reduce these chances. Lower values of HDL cholesterol and increased ratio of total cholesterol to HDL cholesterol are taken as risk factor for CHD.

3. PROTEINS:-


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High albumin levels may be caused by Severe dehydration. o Low albumin levels may be caused by A poor diet (malnutrition). o Severe burns o Kidney/Liver diseases o

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Autoimmune diseases, such as systemic lupus erythematosus (SLE) or rheumatoid arthritis. Gastrointestinal malabsorption syndromes, such as sprue or Crohn's disease. o Hodgkin's lymphoma; Uncontrolled diabetes; Hyperthyroidism; Heart failure. o Decreased synthesis (malnutrition, malabsorption, liver disease, and other chronic o diseases), Increased loss (nephrotic syndrome, many GI conditions, thermal burns, etc.), and o Increased catabolism (thyrotoxicosis, cancer chemotherapy, Cushing's disease, o familial hypoproteinemia). High globulin levels may be caused by Diseases of the blood, such as multiple myeloma, Hod gkin's lymphoma, o leukemia, macroglobulinemia, or hemolytic anemia. Autoimmune diseases, such as rheumatoid arthritis, lupus, autoimmune hepatitis, o or sarcoidosis. Kidney and Liver diseases; Tuberculosis. o o

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Low globulin may be seen in congenital or acquired hypogammaglobulinemic states. o

4. BLOOD UREA NITROGEN:-

Normal Value  

Dog : 12-25 mg/dl Bovine : 20-30 mg/dl


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Decreased level of BUN in Protein malnutrition o Hepatic insufficiency o Increased level of BUN noticed in Physiological factors (Pre Renal ) o High protein diet, dry food and canned food.  Catabolic break down of tissue as consequence of fever ,trauma, infection  and toxemia. Haemorrhage into gastrointestinal tract  Iatrogenically by administration of certain drugs  Anything that reduce renal blood flow 















   

Shock Adrenocortical insufficiency Dehydration Increased protein osmotic pressure

o

Renal

o

Elevation of BUN occur when approximately 70% of nephron are non functional Post Renal Uremia It occurs with obstruction or rupture of excretory pathwa y of urine  ( ureters, bladder and urethra 

5. CREATININE:-


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The increased level of creatinine indicates functional impairment of kidne y. The rate of creatinine excretion is influenced by GFR and any other abnormality that decrease GFR result in an increase in concentration of creatinine. In the earlier stages, Creatinine clearance test is is a sensitive index of impaired renal function. The prerenal factors that increase blood urea have less influences on creatinine concentration. Hence for diagnosis of renal diseases, serum creatinine is preferred over urea estimation. In addition to renal disease, elevated level of se rum creatinine and creatinuria may be observed in extensive muscle extensive muscle destruction. destruction.

6. URIC ACID:-

Clinical Significance  

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In man and all primates, uric acid is the end product of purine metabolism. In plasma at pH 7.4, uric acid is in the ionized form of monosodium/potassium urate, and only a minor portion as a free acid. Its determination help in differentiating gout from non-gout arthritis. Increased levels are observed in gout , leukemia, broncho and lobar pneumonia and polycythemia.

Physiologic changes in serum concentration of uric acid















o

o

o

About 90% of uric acid filtered in glomeruli are normally reabsorbed in renal tubules. Uric acid is normally excreted from the bod y via kidneys (80%) and intestine (20%). About 4.76 mmol (800 mg) are excreted per day. Extreme physical exercise induces a significant increase in the concentration of uric acid. Starvation and fat rich diet increase the concentration of uric acid in serum.

Pathologic changes in serum concentration of uric acid 

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Increased values of uric acid in:

coronary artery disease Down’s syndrome (some cases) gout hyperlipoproteinemia myeloid leukemia pneumonia uremia

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diabetic keto-acidosis following intravenous fructose tissue destruction hemolytic anemias lead poisoning pernicious anemia (especially after treatment)primary and secondary polycythemia

Decreased values of uric acid in acromegaly (some cases) o administration of uricosuric drugs o Fanconi syndrome o hepatolenticular degeneration o xanthiuria. o

7. BILIRUBIN:-

Clinical Significance Determination of bilirubin is of great importance in the differential diagnosis of jaundice. The following types of jaundice are differentiated according to th e concentration of conjugated and unconjugated bilirubin in serum: a. Hemolytic (prehepatic) Hemolytic (prehepatic) jaundice characterized by enhanced hemoglobin breakdown with normal liver function; b. Hepatic (parenchymatous) jaundice due to hepatocyte lesion. Conjugated bilirubin cannot be secreted to biliary capillaries but reenters the blood instead. c. Obstructive (posthepatic) Obstructive (posthepatic) jaundice due to intra- or extrahepatic biliary duct obstruction; and















CLINICAL SIGNIFICANCE &INTERPRETATION OF EXAMINATION OF URINE A. PHYSICAL EXAMINATION OF URINE

1. Colour 

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The colour of the urine may be b e any one of the following: colourless, pale yellow, dark yellow, yellowish-brown, greenish-yellow, red, reddish-brown, brown, green, blue and milky. The yellow colour of the urine is due to urochrome pigment. Interpretation Cloudy - Pus in urine o Dark yellow - Dehydration in vomition, diarrhoea, serous exudations, fever and o deficientintake of water Pale - Diabetes mellitus, diabetes insipidus, increased water intake, pyometra o and chronic interstitial nephritis in dogs Yellowish brown/Greenish yellow - Jaundice o Red - Haemorrhage o Faint pink - Congenital porphyria o Green - Medication with methylene blue or diazan o Brown/coffee coloured - Haemoglobinuria /Myoglobinuria o

2. Transparency 

The transparency of the urine is tested by viewing it against light in a test tube. Normally, it should be transparent and clear. But in the horse, normally, the urine is cloudy cloud y and thick due to the presence of mucus and calcium carbonate crystals. So, the sample of horse urine must be allowed for sometime to settle. The supernatant urine is then used for other tests.

Interpretation 

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The urine is cloudy if it contains epithelial cells in large numbers Leucocytes in large numbers as in pyuria o Large numbers of bacteria o Mucus in the urine of the horse o Crystals-calcium carbonate or amorphous urates or amorphous phosphates o When blood is present, it gives a red or brown smoky appearance o In dogs, if the freshly voided urine is cloud y, then it is pathological due to the presence of pus, blood or mucus.

3. Odour















Interpretation o o o

Sweetish odour in diabetes mellitus In ketonuria, there is odour of acetone. In turpentine medication, the urine may have odour of violet

4. Foam 

Normal urine may foam if it is shaken. In albuminuria foam foam occurs in plenty.

5. Specific gravity 

This is a measure of the relative amounts of solids in solution in the urine. It indicates the capacity of the kidney to concentrate the urine by reabsorption. Generally, specific gravity varies inversely with the quantity of urine. Ex ceptionally, in diabetes mellitus due to the presence of sugar the specific gravity is high though the quantity is also high. Specific gravity is determined by means of urinometer. This con sists of a cylinder and float at the stem of which is scale from top to the neck of the stem. Interpretation Increased specific gravity is seen in:

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Acute interstitial nephritis, the specific gravity is elevated due to the inability of the kidney to excrete water Cystitis due to the presence of products of inflammation Diabetes mellitus due to glycosuria Dehydration, vomition, vomition, diarrhoea and reduced feed intake

Decreased specific gravity is seen in :    

Increased fluid intake Inability of the kidney to concentrate urine Advanced stages of uraemia Diabetes insipidus

















B.CHEMICAL EXAMINATION OF URINE

I. 

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REACTION OF THE URINE

It varies in different species and individuals, depending u pon the diet and metabolism. In herbivores, the reaction is alkaline, where as in c arnivores and omnivores, it is acidic. Horse - alkaline (pH 8) o Cattle - alkaline (pH 7.4 to 8.4) o Sheep - alkaline o Pig - acidic or alkaline o Dog - acidic (pH 6 to 7) o Cat - acidic (pH 6 to 7) o Man - usually acidic (pH 4.8 to 7.5) o Reaction is detected using a litmus paper. Acid urine turns blue litmus red. Alkaline urine turns red litmus blue.

Interpretation 

Acidic urine Normal in carnivorous animals o Nursing calves and foals that are on milk o Diet with an excess of protein o Starvation – Starvation – catabolism catabolism of body proteins o Fever o Acidosis – Acidosis – both both metabolic and respiratory o Diabetes mellitus  Uraemia  Prolonged muscular activity o Administration of acid salts o Sodium acid phosphate  Ammonium chloride  Sodium chloride 

















Urine becomes alkaline when the sample is ke pt at room temperature because of ammonia formation as a result of decomposition of urea.

o

II.

PROTEIN

Normal urine does not contain any protein Interpretation

Physiological proteinuria 

It is transient and believed to be due to a temporary increased glomerular permeability as a result of capillary congestion. Excessive muscular exertion o Convulsion o Emotional stress o Ingestion of an excessive amount of protein o

Pathological proteinuria 

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Pre-renal: The protein originates from non-renal conditions and its loss in the urine is not due to primary renal disease. Bence – Bence – Jone’s Jone’s protein o Haemoglobinuria o Myoglobinuria o Renal Increased permeability of glomerulus o Impaired reabsorption of protein normally present in glomerular filtrate due to o tubular disease

















Neoplasms and emboli III.

GLUCOSE

Normal urine does not contain glucose. Although glucose passes through the glomerulus, it should be reabsorbed by the proximal convoluted tubules. Interpretation Interpretation - Glycosuria Emotional stress Diabetes mellitus o Hyperthyroidism o Acute and chronic pancreatic necrosis o Hyperpituitarism o Overactivity of adrenal cortex o Shock o Chronic liver disease o Enterotoxaemia in sheep o Brain tumors o Rabies o False positive reaction for glucose can occur if oth er reducing agents like antibiotics, lactose, ascorbic acid, salicylates, morphine, formaldehyde and uric acid are present. o

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IV.

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KETONE BODIES

Ketone bodies are acetoacetic acid, beta-hydroxy butyric acid and acetone. They are not normally present in the urine. But, when fat metabolism is impaired, ketone bodies accumulate in the body, giving rise to ketonaemia and then ketonuria results. Interpretation

















Haemoglobin contains a peroxidase which can liberate oxygen from hydrogen peroxide. This oxygen produces blue colour in the presence of benzidine Interpretation Haematuria Acute nephritis Nephrosis – Nephrosis – marked marked degeneration o Renal infarction o Passive congestion of kidneys o Neoplasms of kidney, bladder or prostate o Urolithiasis – Urolithiasis – renal, renal, cystic or urethral o Abscess of kidney o Pyelonephritis o Ureteritis o Cystitis o Trauma to urethra – urethra – usually usually from improper catheterization o During estrus or postpartum in female due to contamination b y uterine or vaginal o discharge Severe infection – infection – anthrax, anthrax, leptospirosis, infectious canine hepatitis o Chemical poisonings – poisonings – Copper, Copper, Mercury, Sulfonamides and Phenol. o Thrombocytopaenia o Sweet clover poisoning o Parasites - Dioctophymarenale - Dioctophymarenale and Dirofilariaimmitis and Dirofilariaimmitis in in canines o Acute vegetative endocarditis and congestive heart failure in canine o If the blood is seen in the last drop of urine, then the source is bladder. blad der. If the urine is red throughout, the source of blood is kidney. kidne y. If the first portion of urine is red, then the source of blood is some urethral lesions. o

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Haemoglobinuria

















VI.

BILE PIGMENTS

Normally, bilirubin is present in freshly freshly voided urine. On standing, it is converted into biliverdin (green colour), bilicyanine (blue) and cholestelin (violet, red, reddish yellow). Interpretation Moderate to severe hepatocellular damage Obstruction of bile ducts, extrahepatic or intrahepatic o In early hepatocellular damage and haemolysis urine bilirubin may be negative. o

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VII.

UROBILINOGEN

Urobilinogen is normally present in urine. But, it will not be present in the case of obstructive jaundice. Interpretation 

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Decreased amount or absence of urine urobilinogen Obstruction of biliary passages o Decreased destruction of erythrocytes o Impaired intestinal absorption o Nephritis o Increased amount of urine urobilinogen Hepatitis o Cirrhosis of liver o Haemolytic jaundice o



















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Laboratory evaluation evaluation & diagnosis of samples for parasitic diseases

Practical no.1 

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PARASITES / SEGMENTS OF HELMINTHS

Collection

Collect the helminths during postmortem examination, sometimes immature amphistomes and segments of tape worms can be collected from dung or faeces. 

Preservation

Trematodes: Fresh Trematodes: Fresh and live flukes are collected in normal saline. 70% or 90% ethyl alcohol used as a preservative and fixative. Nematodes: Nematodes Nematodes: Nematodes are fixed in 5-10% hot formalin or 70-90% ethyl alcohol. Once the nematodes are fixed they can be stored indefinitely in glycerin alcohol. Cestodes: Tapeworms Cestodes: Tapeworms including the scolices (heads) should be placed in water at about 37ºC for about one hour and then stored in a mixture of 5% glycerine and 70% alcohol or 5-10% formalin.

A. DIRECT EXAMINATION



















Smear the solution over the slide into a thin film Place a cover slip over the smear Examine the area of the slide under cover slip with the compound microscope. The slide is investigated using a magnification of 10x and 40x. Me M eri ts

This method is quick. Minimal equipment is needed. D emer i ts

It is effective only when the concentration o f parasite stages is high. It is frequently difficult to identify them since they are partially covered by debris. Quantitative results cannot be obtained. II.

Stained smear method

a. Iodine violet mount b.Lactophenol cotton blue method 

Parasites found with the direct smear method :

Coccidia and helminth eggs (only when wh en high numbers are present); Cestodes and trematode e ggs



















Be sure the density of the mount moun t is correct. You should be able to read small print through it (but not too clearly). If it is too thick or too thin, observation of the elements in the mount may be difficult. B.SEDIMENTATION TECHNIQUE Procedure: -

Place a lump of faeces (1/2-1 (1/2 -1 tsp / 5-10 g/ 5-10 faecal pellets p ellets (of sheep or goats) in a cup or o r glass container. Add enough tap water and an d mix thoroughly with a spatula / glass rod until all the faecal material is broken down. The mixture is poured through a wire mesh sieve to remove coarse large lumps. Common tea strainer will do. The strained fluid is collected in a bowl. The sieve is rinsed with water and the debris left on the sieve is discarded. Transfer the suspension to centrifuge tubes and ce ntrifuge at 2000 rpm for 2 min. Discard the supernatant. Mix the sediment well and take a small quantity of it and mix it with a drop of water on a clean



















Eimeria oocysts Merits

This is one technique by which the trematode eggs can be recovered. Operculated eggs like that of Diphyllobothrium of Diphyllobothrium latum also latum also will be sedimented. Demerits

Usually the lighter eggs of nematodes and oocysts o ocysts of protozoa are missed while employing this technique. Hence it is not considered by many people as suitable for these types of parasites. The presence of debris which may pose a problem whie doing examination.

S.

Species

Host

No

Description of eggs

. 1

Catttle,sheep,goat,Hors Oval shape, e,

thick walled, presence of



















3 Trichuris sp

sheep,

Barrel shaped

goat, cattle,pig,dog

with a conspicuous plug at both ends,yellow or brown colour

4

Similar to Trichuris sp Trichuris sp but with truncated plugs

Capillaria sp Capillaria sp

Interpretations





















The top film can then be removed and examined. Many nematode and a few cestode eggs float in a liquid with a specific gravity of between 1.10 and 1.20. Trematode eggs, which are much heavier, require a specific gravity of 1.30-1.35. I.

SIMPLE FLOATATION TECHNIQUE

Simple floatation / Will’s technique Small floatation tube with emulsion of faeces in the above solutions is filled to its full capacity till a convex surface is formed and is allowed to stand for 20 to 30 minutes, by which time eggs would have floated up. Apply a coverslip or slide to the surface to remove the first drop of fluid, containing e ggs and then examine that drop on the slide.





















Practical no.2 QUANTITATIVE FAECAL EXAMINATION 

The estimation of the worm burden or oocyst load can be done by quantitative methods of faecal examination. This is useful in knowing the worm burden, epidemiology of an infection and to note the effect of treatment.

Advantage 

Quantitative FE (faecal egg count) is essential for comparing the e fficacy





















MC MASTER METHOD 

This technique is used for counting of eggs, e ggs, larvae or oocysts, which float in a saturated solution of sodium chloride.

Procedure 

Weigh 1 g of faeces and add 14 ml tap water.

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Homogenize and pour suspension through a 250 µm aperture sieve, collecting the filtrate.

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Collect the filtrate, agitate and fill a test tube of 15 ml volume. Centrifuge at 2000 rpm for 2 min.





















Bunostomum sp.

300-700

50-100

Haemonchus sp.

2500-8000

200-700

Ostertagia sp., Trichostrongylus sp.

250-2000

100-500

Nematodirus sp.

100-600

50-100

Fasciola sp.

200-500

10-25

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The data is circumstantial and subject to factors such as a ge, sex and breed of animals, immunological status, feeding conditions, etc.





















Practical no.3 PASTURE LARVAL COUNTS&SKIN SCRAPPING EXAMINATION

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Introduction

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It is a useful technique for detecting the level of infective larvae (L3) of gastrointestinal nematodes on pastures. It is not a clinical laboratory technique.

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Collection of materials

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Collect a reasonable number of samples of grass from the pasture and place in polythene























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The fluid is then centrifuged at 1000 rpm for 5 minutes to concentrate mites, which can be collected from the sediment.

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Discard the supernatent, collect a drop of the sediment, transfer to a slide and examine for























3

Sarcoptes

Dog, cat rabbit,

Body is globose and striate with scaly and spinose areas,

spp

pig, cattle,

posterior pairs of legs short, not extending beyond body

sheep, goat

margin,pedicels bear sucker or bristle and not segmented,























Practical no.4 EXAMINATION OF BLOOD FOR PARASITES























Chylomicrons and superimposed platelets appear as bright, translucent objects lodged above or below RBC’s. They tend to get separated on constant observation due to Brownian movement in wet medium and are extra cellular.























While Leucocytozoon While Leucocytozoon,, microfilariae and sometimes Trypanosoma can Trypanosoma can be found with the lower powers of the microscope, the stained smears should be examined with the oil immersion objective for other protozoa.











































































































































































































































































































































































































































































































































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