Midterm II Flashcards
What faeces tests are useful for EPI determination/diagnosis?
Dyed agar-gel digestion and Schwachmann-filmtest- uncharged x-ray film contains gelatin + pH increaser + feces and incubate. After 30-60 minutes if x-ray film becomes transparent there is digestive enzymes in the feces. You can also stain agar and be prepared so it has a stable pH value of 8, pour into petri dish. Place piece of fecal sample into middle of dish after 30-60minutes we can see a wide digested circle around sample if feces contains enough digestive enzymes.
Smears to detect nutrients. Giemsa- undigested fibers.
Lugol- undigested starch particles
Sudan III- undigested lipid particles
How can urine specific gravity be measured? When is the urine hypo normo and hyperstenuric?
refers to urine osmolality
indicator of concentrating ability (tubular function). SG is the ratio of the weight of the liquid to an equal volume of distilled water. SG of urine is always greater than distilled water. SG increases with increasing concentration of dissolved ions (osmolality), glucose, proteins, lipids and contrast materials.
urinometer – best method
refractometer – problem if lot of glucose or if urine is not homogenous
test strip – not so good test
-Hyposthenuria: SG<1.008 Temporary hyposthenuria can be normal with an increase in water uptake but if it is persistent then it can be pathological. causes: Hyperadrenocorticism, liver disease, prolonged fluid therapy.
-Isothenuria: SG 1.008-1.012. Temporary can be normal due to increased water uptake, Persistent can be pathological. Indicator for severe tubular damage.
-Hypersthenuria: SG >1.012 (Normal). as major function of tubules is to concentrate the primary glomerular filtrate. Can be pathological if 1) decreased water intake 2) substantial water loss 3) acute kidney failure. SG may be increased in diabetes mellitus due to increased glucose concentration.
SG is important in polyuria and polydipsia.
What is ALT?
Alanine Aminotransferase. In cytoplasm of liver and red blood cells. better sensitivity and specific than AST . Converts alpha-keto-glutaric acid to L-glutamic acid and L-alanine to pyruvic acid. Carnivore liver specific. normal ca >60 IU/l Increase: liver cell damage chronic active hepatitis cirrhosis bile duct obstruction liver neoplasm pancreatitis septicaemia copper storage disorder drugs – barbiturates, glycocorticoids, salicylates
Proximal and distal ileus
PROXIMAL ILEUS foreign body in stomach or prox. duodenal part. vomit that contains HCl early on metabolic alkalosis later metabolic acidosis dehydration anaerobic glycolysis
DISTAL ILEUS foreign body in jejunum and more distal parts animal don’t eat/empty stomach dehydration anaerobic glycolysis lactic acid formation metabolic acidosis alkaline vomit
List and group the liver enzymes. What information is gained about the liver in case of enzyme deviations
PARENCHYMAL ENZYMES aspartate aminotransferase AST alanine aminotransferase ALT glutamate dehydrogenase GLDH sorbite dehydrogenase SDH
BILE DUCT OBSTRUCTION ENZYMES
cholestatic: ap-alkaline phosphatase ALKP
gamma glutamyl-transferase GGT
measured by kinetic spectrophotometry
not parameters for liver function but damage
What is AST, where is it found? Where can we see it‘s elevation in the plasma? In what species is it useful?
AST – ASPARTATE AMINOTRANSFERASE in mitochondria of liver, muscle, red blood cell converts alpha-keto-glutaric acid to L-glutamic acid & L-aspartate to oxalic acetic acid herbivore liver specific normal dog >30 IU/l Increase: hepatitis fatty liver neoplasm hemolysis muscular injury training myocarditis herbivores: hepatopathy ca: severe parenchymal damage
What is AP?
ALKP – AP-ALKALINE PHOSPHATASE
in cytoplasm, produced by different organs (biliary epithelial cells, hepatocytes, bone)
only hepatic (heat-atable) & bone (heat-labile) appears in blood
optimal pH 10
phosphotransferases – phosphate-esters are translocated to alcohol or phenol
hydroxyl group to another
normal ca/ru >200 IU/l (not used in cat because of its short half-life)
Increase: hepatitis
cholestasis
acute hepatic necrosis
cirrhosis
neoplasm
internal/external biliary obstruction
steroid induced hyperadrenocorticism (SIAP)
drugs – barbiturates, glycocorticoids, salicylates
What is GGT?
GGT – GAMMA GLUTAMYL-TRANSFERASE
produced by different organs, liver origin appears in blood
L-alpha-AA can be better attached to reduced glutathione & transported through
membranes
normal ca >10 IU/l (good parameter for cat & eq)
Increase: hepatitis
cholestasis
acute hepatic necrosis
cirrhosis
neoplasm
internal/external biliary obstruction
steroid induced hyperadrenocorticism (SIAP)
drugs – barbiturates, glycocorticoids, salicylates
Detecting protein from urine?
PROTEINURIA
very low, measured by ultrasensitive protein measurement
proteinuria (mainly albuminuria) is an essential indicator for glomerular dysfunction
increased cc – proteinuria febrilis
proteinuria/day <20 mg/kg bw/day
test strip – AA of proteins cause a colour change on test strip blue-greenish. Most sensitive to albumin.
sulfosalicylic test – 4-5 drops of acid to 1-2ml of sample. Sample becomes more opaque if more coagulated precipitates are formed due to higher protein cc. More sensitive than tests strips, but not specific.
gmellin (heller) test – Showing bile pigment metabolites and protein content of the urine. ccHNO3 layered underneath the urine in a TT. white ring of denaturated proteins.
Spectrophotometric method- methods can be grouped 1) colour intensity of a complex formed by direct reaction of urinary proteins with dye-reagent. 2) have to concentrate the protein(less practical) .
Urine protein: Creatinine protein
Determination of Bence-Jones proteins
List the laboratory tests which allow us to evaluate the impairment of glomerulus-function.
1) Blood Urea (BUN=blood urea nitrogen) concentration in blood plasma
2) Creatinine concentration in blood plasma
3) Plasma urea (mmol/L) / Plasma creatinine (μmol/L)
4) Creatinine clearance
5) Urinary Total Protein conc.
6) Urinary Total Protein / Urinary creatinine ratio
7) others: Radioisotopic methods, C-inulin clearance, H-tetraethyl-ammonium-chloride clearance.
What are the causes of increased blood urea concentration?
-PRERENAL decreased perfusion (shock, dehydration) increased nitrogen/protein intake energy deficiency in rumen- ruminal microorganisms can not produce enough protein so catabolised protein (NH3) is absorbed intestinal protein catabolism intestinal or gastric bleeding hemolysis fever liver function Decreased blood perfusion in kidneys (Addisons) -RENAL reabsorption uremia (tubular obstruction) retention uremia (glomeruli problem) -POSTRENAL inhibition of urine flow reabsorption uremia (lower urinary tract rapture, obstruction)
What are the causes of increased blood creatinine concentration?
state of muscles (acute trama/injury, necrosis, cachexia) kidney glomerulus function: -prerenal decreased perfusion nitrogen/protein intake energy deficiency intestinal protein catabolis intestinal or gastric bleeding hemolysis fever liver function -renal neoplastic process retention uremia -Post renal inhibition of urine flow reabsorption uremia (lower urinary tract rupture)
What is proteinuria? How can it be evaluated?
Normally, primary ultrafiltrate contains 20-30% proteins and it is reabsorbed by tubules almost completely, thus protein content of normal urine is very low. measured by ultrasensitive protein measurement
proteinuria (mainly albuminuria) is an essential indicator for glomerular dysfunction
increased cc – proteinuria febrilis
proteinuria/day <20 mg/kg bw/day
test strip – not good, react mainly with albumin
sulfo test – denaturation of proteins, becomes whitish
gmellin (heller) test – white ring of denaturated proteins
What info is provided by the urine TP/creatinine ratio? Where can it be used
When high total protein cc is measured in urine, we measure this ratio as it is not influenced by SG of urine
physiological value <0.8, if 1-5 the proteinuria is prerenal cause, if >5 renal disease
Water deprivation test - What is the goal of this test, its method and interpretation.
preformed very rarely! used to evaluate the causes of polyuria and polydipsia.
used to detect diabetes insipidus.
needed parameters – bw, Ht, creatinine, urea, TP, osmolality
empty bladder, weigh patient and urine sampling repeat every hour
water is withheld until bw is decreased by 5% from basal weight-usually 6 hour deprivation is needed. Urine specific gravity should be measured at this time.
Results: 1) 1050-1080g/l: psychogenous polydysia
2) 1001-1007g/l: tubular cell damage or decreased ADH function. Give ADH IM or Intranasally and check SG after 60 minutes, a) 1050-1080g/l: Central diabetes insipidus (decreased ADH production). b) 1001-1007g/l: peripheral (renal) diabetes insipidus (decreased effect of ADH on damaged tubular cells)
What is the goal of Enzymuria Evaluation? Goal and how is it performed/tested?
for tubular function tubular cells contain ALKP & GGT Their release into URUNE is increased in acute/peracute tubular damage. Values must be referred to the creatinine levels in order to exclude misdiagnosis caused by high enzyme level in concentrated urine. Alkaline phosphatase (U/l)/ Creatinine (umol/L): normal=0.02 Gamma-Glutamyl transferase (U/I)/Creatinine (umol/L): normal=0.01
List the causes of abnormal urine pH (increased, decreased).
ca – acidic (65.5-7.5)
herbivores – alkaline (7-8.5)
postprandial alkalization (kidney compensation)
food changes pH slightly within physiological range
test strip – only whole numbers
test paper
Increase-Alkaline: 1) Feeding in carnivores results in alkalisation of urine. highly acidic gastric juice is released in the stomach after feeding resulting in slight metabolic alkalosis.
2)Urinary tract infection (UTI) caused by urease producing bacteria. ureases enzyme breaks down urea to ammonia-increasing pH.
3) Metabolic and respiratory alkalosis result in decreased H+ excretion (increased alkaline intake, increased loss of C02)
4) proximal renal tubular acidosis causes an increased HCO3- excretion to urine
5) long storage tome causes urea decompensation to ammonia (pH increase), can also occur in urinary tract obstruction.
Decrease-Acidic: 1) Metabolic and respiratory acidosis- increase of H+ excretion in the kidney (HCO3 loss due to diarrhoea, ileus, muscle weakness)
2) vomiting, Na+ is reabsorbed with HCO3-, due to decreased HCO3- excretion in urine pH becomes more acidic
3) hypokalaemia there is increased H+ excretion in exchange to reabsorbed K+
4) treatment with acidic drugs
5) distalis renalis tubularis acidosis
6) abomasal displacement
7) toxicosis with acidifying substances (metaldehyde)
What can be seen in the physiological urine sediment? List the pathological abnormalities in the urine sediment.
Organic sediment: Blood cells(WBCs and RBCs) abnormal in haematuria: renal or urinary tact disease, obstruction, trauma
Cells from the lower urogenital tract: abnormal- inflammation or infection of urinary tract
Viral inclusion bodies: distemper
Microbes : bacteria
Mucin
Casts(cylinders): Abnormal- increase- acute tubular disease or renal tubular damage, haemorrhage(RBC cast), inflammation(WBC cast)
Inorganic sediment: small amount of struvite or different calcium- crystals may be found in normal urine. Larger number lead to stone formation.
Mostly in alkalic urine:
Struvite: UTI
Calcium Carbonate: hypercaluria
Calcium-Phosphate: hypercalcuria
Amorphous Phosphate: Meat and grain diet
Ammonium-ureate/biurate: severe impairment of hepatic function in Dalmatian dogs
Mostly in acidic urine:
Calcium oxalate: toxic plant consumption
Uric acid: Dalmations
Cystine, Tyrosin, Leucine: metabolic disease
Bilirubin: prehepatic and hepatic jaundice
Sulphonamides: sulphonamide therapy
How can the presence of blood or haemoglobin be shown in the urine? How can the two be distinguished from each other
Haematuria is the presence of blood(intact RBC) in urine. Haemoglobinuria and Myoglobinuria is presence of free dissolved haemoglobin and myoglobin in urine. If presence is seen by naked eye colour of urine- pink, red, brownish.
benzidine test – sample into reagent, check pseudoperoxidase activity. Not specific test.
test strips: speckled appearance of test pad-haematuria, diffuse colour- haemogloburia or myoglobinuria.
Urine sediment analysis: RBCs can be seen microscopically.
Haematuria can be differentiated by centrifuging the sample, since RBCs sediment, the supernant clears up, in haemoglobinuria and myoglobinuria the urine supernant remains reddish. While in haemoglobinuria usually concurrent reddish discolouration of plasma and anaemia are also observed, in myoglobinuria the plasma is clear and muscle damage indicator enzyme activities are elevated.
Causes of haematuria:
lower urinary tract infection, trauma, genital tract injury, tumor or inflammation
Causes of hemoglobinuria:
intravascular haemolysis
long term stasis of blood in urinary bladder
Causes of myoglobinuria
excessive muscle trauma or exercise, burns
DISTINGUISH HEMOGLOBULINURIA FROM MYOGLOBULINURIA
creatine-kinase test
lactate dehydrogenase test
aspartate aminotransferase test
How can the presence of pus be shown in the urine? What are the causes?
pus: an accumulation of neutrophil granulocytes, some tissue cells and microbes. Pus in urine-pyuria. microscope – sediment evaluation donne test – reagent makes sample more viscous if pus is present (and so bubbles elevate slower). (-, +, ++, +++) Positive result may occur in males if sperm is present. Causes: Kidney pelvis inflammation cystitis (inflammation in urine bladder) inflammation of penis/vagina: endometritis (females) prostatitis (males)
Hapatocellular enzymes in dogs
ALT, GLDH
What is TLI? Describe its usefulness in the diagnosis of EPI
TRYPSIN LIKE IMMUNOREACTIVITY
Tests for pancreatitis and EPI
most specific method.
Determined by radioimmunoassay (RIA) and ELISA
antibodies are produced against one part of trypsinogen. Antibodies marked by radioisoptopes are bound to trypsinogen in sample. marked trypsinogen cause increased radioactivity that can be measured.
trypsinogen goes to duodenum & is activated by enterokinase to trypsin this can get into blood stream and the cc will be 15 times more than normal.
If TLI-cc is less than 2.5 ug/L the probability of EPI of decreased enzyme synthesis origin is reliable. if EPI is caused by obstruction of the pancreatic duct, TLI level is normal or high.
Describe the Gmelin-Test.
C.c. HNO3 has to be carefully layered under urine in a test tube, and the width of the differently coloured layers at the meeting of the two fluid phases has to be evaluated. Differently coloured layers from the highest layer: condensed material on the surface of the glass tube - acidic urea yellow - urine itself white (opaque) – protein purple - indicane (indol-sulphate) green – biliverdin brown - urobilinogen (UBG) HNO3
List the most important findings in the blood and urine for prehepatic icterus.
Blood: Br I ↑↑ Br II ↑ UBG ↑↑ Free Haemoglobin ↑ Haptoglobin cc. ↓ Ht ↓ Reticulocyte count↑↑↑ Coag. parameters (PT, APTT, TT) ↔↑(DIC) enzyme activities: ALT, LDH↑ substrate cc: Urea↑ Urine: UBG↑↑ Br↑ Hb↑↑↑
List the most important findings in the blood and urine for hepatic icterus.
Blood: Br I ↑ Br II ↑↑ UBG ↑ Free Haemoglobin↔ ↑ Haptoglobin cc. ↔ Ht ↔↓ Reticulocyte count ↔↓ Coag. parameters (PT, APTT, TT) (no change after vitamin K administration) enzyme activities: ALT, AST, GLDH↑ substrate cc: Urea↓ , NH3, Bile Acids↑ Urine: UBG↑ Br↑↑
List the most important findings in the blood and urine for posthepatic icterus.
Blood: Br I ↔↑ Br II ↑↑↑ Free Haemoglobin↔ Haptoglobin cc. ↔ Ht ↔ Reticulocyte count ↔ Coag. parameters (PT, APTT, TT) (improves after vitamin K administration) enzyme activities: ALKP, GGT↑ substrate cc: Bile acids↑ Urine: UBG↓ Br↑↑↑
Bile acids. How to measure them and when is their concentration elevated in the serum/blood?
Bile acids (cholic acid, chenodeoxycholic acid) are synthesized in the liver from cholesterol. Measurement: HPLC, RIA or total bile acids (TBA) There are two ways of sampling: 1) after 12 hours starvation, 2) after eating, postprandial value - postprandial value can be 10 times (dog), 5-6 times (cat) more than the value measured after starvation. Na-EDTA or -citrate can be used as an anticoagulant, because heparin disturbs the measurement. Normal value (fasting): 6 µmol/l (carnivores) 20-30 µmol/l (other animals).
Increased:Liver injury, hepatic cell damage - increased outflow of bile acids from the damaged hepatocytes to the blood, bile duct obstruction or bile endothelial cell damage - decreased secretion of bile acids to the bile, increased outflow to the plasma instead, decrease in liver function, therefore decreased uptake of the absorbed bile acids (note: increased urobilinogen level !) biliary stasis (cholangiohepatitis cirrhosis, hepatic or pancreatic neoplasm, pancreatitis) portosystemic shunt (absorbed bile acids bypass liver tissue)
What method is used in blood ammonia concentration measurement? In which diseases is blood ammonia concentration increased?
produced in intestine by bacteria, absorbed & detoxified in liver by urea cycle
cc measurement = liver function test
starve animal 24 hours, avoid air contamination when sampling,
spectrophotometry or refractometry
normal dog 26.4-70.5 micromol/l
Ammonia checkers
Ammonia tolerance test: after 24 hours of starvation and pretreatmen tof neomycin, take basal blood sample and give NH4CL in 5% water solution, blood samples taken every 30-40 minutes. if liver function is normal, ammonia cc is <120umol/L in dogs and <175 umol /L in cats.
INCREASED AMMONIA CONCENTRATION
impaired liver function – decreased urea production, liver cirrhosis, portosystemic
shunt, liver lipidosis, hepatitis, toxic damage
ruminal alkalosis/ammonia toxicosis
ru/eq/rabbit – intestinal overgrowth of NH 3 -producing bacteria
congenital enzymopathies: decreased activity
What are the causes of proteinuria vera and proteinuria spuria?
True proteinuria (proteinuria vera): proteins from or through kidney kidney disease sudden protein catabolism (fever, hemolysis, exercise, starvation, onset of glucocorticoid treatment)
False proteinuria (proteinuria spuria) proteins from lower urinary tract or genital tract lower urinary tract infection tumors prostatic testicular inflammation
Rivalta test- goal, how to perform, interpretation
Prepare 3% acetic-acid solution. One-two drops of native samples (NOT CENTRIFUGED) dripped into the solution. If there is coagulation(labile proteins-fibrinogen, globulins) there will be a smoky appearance, this result is exudative. (+/++/+++)
If coagulation is not observed but sample is dissolved in the solution the result is transudate(stabile proteins-albumin) stabile proteins do not show coagulation with weak acids (-).
If sample contains a lot of globulins the sample goes honey like in case of FIP after adding some drops of sample into solution the smoky appearance of coagulation is not visible., but sample does not dissolve in solution but remains forming one drop. In this case globulins are coagulated on the surface.
When does alpha-amylase activity increase?
acute pancreatitis acute, subacute kidney failure FIP & other immune-mediated diseases lymphoma, myeloma diabetes mellitus ileus gastric or intestinal perforation parotitis chronic enteritis
EPI. What is it, how to test it? (Laboratory diagnosis of EPI)
Exocrine pancreatic insufficiency Develop due to chronic necrotic or atrophic damage to the pancreas., or inherited disease. Features: - decreased production of digestive enzymes or enzymes do not get out from organ to intestines. - Maldigestion - pancreatogenous dyspepsia - weightloss - overeating & vomit - coprophagia - allotriophagia - Large amount undigested food in intestines cause bacterial overgrowth - Intermittent chronic diarrhea - bacterial enteritis - frequent flatulence - undigested fat on surface of feces - vitamin deficiency - skin an fur problems - mild anemia. Laboratory examinations: - TLI concentration - BT-PABA test - Dyes agar-gel digestion and schwamann filmtest - Lipid absorption test - Fecal elastase test
Biochemical parameters for examination of free fluids.
- outlook physical parameters (colour, odour, consistency)
- Rivalta-test
- coagulation ability
- specific gravity
- pH
- nucleated cell count
- total protein concentration
- albumin/globulin ratio
- creatinine, urea concentration
- alpha-amylase, lipase activity
- LDH activity
- triglycerol, cholesterol concentration
- cytological analysis
What is the lipid absorption test, and which disorders can it confirm or eliminate
perform this test in order to determine whether there is existing lipid malabsorption, maldigestion , especially in EPI or chronic small bowel disease . When there is fast lipid intake in normal conditions, plasma triglycerol (TG) concentration rises to twice as the normal value (normal for dogs: 1 mmol/l).
If the result is increased TG concentration and lipaemia, we can suspect that the original problem is exocrine pancreatic insufficiency (EPI). If the result is no change in TG concentration and lipaemia is not seen, we can suspect intestinal absorption defect
Laboratory diagnosis of pancreatitis
urine amylase: ↑ urine creatinin: ↑ pancreatitis plasma amylase:↑
plasma creatinin:↔(↑)
ALT-level in blood
normal ca >60 IU/l
Causes for high or low blood-creatinine concentration.
high: inflammation of muscles muscle trauma Increase meat intake Low/normal: cachexia chronic muscle atrophy
Urea conc. in blood
from liver urea cycle (detoxified NH 3 absorbed from intestines) nontoxic but osmotically active filtrated through glomeruli reabsorbed from tubules normal 8-10 mmol/l
Urinalysis- Main test
Urinalysis is a basic diagnostic tool like haematology and biochemical profile of the blood. Urinalysis is the physical, chemical, and microscopic examination of urine.
Free catch sample - use a clean or sterile container and avoid contamination of the sample with hair, dust or mud etc, catch urine middle stream for most reliable results.
Liver function test
PT, APTT, TT (not specific)
Albumin (not very sensitive)
Bilirubin (not specific)
Bile acids (best liver test, sensitive but not very specific) Ammonia (conversion of ammonia to urea occur in liver)
Proteinuria: causes and types
Causes of proteinuria Proteinuria may be caused by glomerulonephropathy, tubular transport defect, inflammation, infection or marked haematuria within the urogenital tract. Considering the causes of proteinuria it is important to distinguish true or real proteinuria and false proteinuria.
- Real Proteinuria i.i. nephrongenic (proteinuria vera)
a. Selective proteinuria
b. Non-selective proteinuria - Pseudo-Proteinuria (spuria)
Local and general consequence of distal ileus
local anaerobic glycolysis & lactic acidosis →tissue necrosis & inflammatory processes
fluid accumulation prox to intestinal block
bacterial overgrowth due to content accumulation
gram – release endotoxins (absorbed → endotoxaemia, shock)
gram + release exotoxins (absorbed→ bacteraemia, sepsis)
Ht (PCV) increased
glomerular filtration rate & renal function decreased
hypokalaemia
hyperkapnia, hypoxaemia cause respiratory acidosis
painful abdominal cavity→ stress & adrenaline effect
bacteria & intestinal hypoxia → can cause pancreatitis & liver damage
Causes of increased ALKP in the blood
hepatitis cholestasis acute hepatic necrosis cirrhosis neoplasm internal/external biliary obstruction steroid induced hyperadrenocorticism (SIAP) drugs – barbiturates, glycocorticoids, salicylates
Causes of decreased urine pH
metabolic/respiratory acidosis
distal tubular defect (decreased HCO 3 excretion)
fanconi syndrome
hyperkalaemia + metabolic alkalosis (paradox aciduria)
toxicosis
acidotic substances (eg. ammoniumchloride)
abomasal displacement
Inorganic components of urine and one example of why they would be found in urine.
a small amount of struvite and calcium crystals can be found in urine but are increased when there are a large number it can lead to stone formation. Urinary Tract Infection (UTI) can also cause an increase of these in the urine.
