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LIVER AND KIDNEY FUNCTION PARAMETERS IN AVIAN SPECIES. A REVIEW Ebeid,T. A.; Eid,Y. Z. And El-Habbak, M. M. Dept. of poult. Prod. , Kafr El-sheikh Fac. of Agri., Tanta Univ. , 33516 Kafr El-sheikh, EGYPT. Abstract: There is a great deal of research and literature about liver and kidney function including the clinical chemistry of different avian species. This paper provides a review of different parameters used for evaluating the functional status of liver and kidney. Moreover, interpretation of the results of standard biochemical analyses, including alanine aminotransferase, aspartate aminotransferase, gamma glutamyltransferase, bilirubin, ammonia, alkaline phosphatase, lactate dehydrogenase, cholesterol, glucose, total protein, albumin, globulins, calcium, phosphorus, antioxidative enzymes and lipid peroxidation is reviewed and discussed in relation to these physiological differences. The use and interpretation of alternative analyses appropriate for avian species, such as uric acid, biliverdin, creatinine, creatinine clearance, inulin clearance, uric acid clearance and urea clearance also are reviewed. This literature review could answer some questions for researchers, and hopefully, it is used as a guide for interpretation of their results. Key words: alanine aminotransferase, aspartate aminotransferase, gamma glutamyltransferase, bilirubin, ammonia, alkaline phosphatase, lactate dehydrogenase, cholesterol, glucose, total protein, albumin, globulins, calcium, phosphorus, antioxidative enzymes.

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INTRODUCTION The liver plays a major role in metabolism and has a number of functions in the body including glycogen storage, plasma protein synthesis and the formation of uric acid. It is a hematopietic organ during the embryonic and immediate postembryonic periods. (Sturkie, 1976; Ziswiler and Ferner, 1972). Also, the liver is involved in the detoxication of metabolities, and it extracts sodium bromsulphthalein (BSP) from the blood and excretes it in the bile (Sturkie, 1976). The kidney performs many physiological and excretory functions. It performs three main functions: filtration, excretion or secretion, and absorption. It filters water and some substances normally used by the body from the blood, along with waste products of metabolism, which are voided in the urine. It conserves needed body water, glucose and other substances by reabsorption. These processes make the kidney an important homeostatic mechanism whereby the body water and solutes are maintained at fairly constant levels. Filtration takes place in the glomeruli whereas secretion and reabsorption take place in renal tubules (Sturkie, 1976; Kaplan and Pesce, 1989; Ganong, 1997). The avian kidney filters a large volume of fluid (approximately 11 times the entire body water each day for a 100-g bird) and then reclaims most filtered water by tubular reabsorption (Goldstein and Skadhauge, 2000). The main objective of this article is to consider the different parameters used for evaluating the functional status of liver and kidney and interpretation of the results of standard biochemical analyses. LIVER FUNCTION TESTS The liver is an organ of many diverse metabolic activities, and any assessment of its functional status is dependent upon its ability to perform a specific metabolic function. A number of tests have been devised for the detection of alterations in liver function. All liver function tests may be classified according to the type of hepatic function examined. According to Coles (1986a) liver function tests may be categorized as follows: 1. Tests dependent primarily on hepatic secretion and excretion. A. Bile pigments. B. Clearance of foreign substances.

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2. Tests dependent upon specific biochemical functions. A. Protein metabolism tests. B. Carbohydrate metabolism tests. C. Lipid metabolism tests. 3. Tests dependent upon the measurement of serum enzyme activity. A. Transaminases. B. Alkaline phosphatase. C. Other enzymes. First: Tests Based on Hepatic Secretions and Excretions Bile Pigments Serum Bilirubin and Biliverdin: It is generally considered that biliverdin is the primary end product of heme catabolism and the principal bile pigment of the domestic fowl (Sturkie, 1976; Cornelius, 1981). Biliverdin is excreted into the bile as a bile acid complex of sodium biliverdinate in chickens and turkeys. Birds lack the enzyme biliverdin reductase needed to reduce biliverdin to bilirubin. Therefore, bilirubin accounts for only a small percentage of the total bile pigment in birds that have been studied (Lin et al., 1974). Biliverdin (the tetrapyrrole dehydrobilirubin) can be measured in research laboratories by high-performance liquid chromatography (HPLC) (Hornbuckle and Tennant, 1997). Little, if any, biliverdin or bilirubin is detectable in the plasma of normal birds. Bile obtained from the hepatoenteric duct had 70 percent more bilirubin than biliverdin. Ligation of one bile duct was followed by no hyperbiliverdinemia and slight hyperbilirubinemia. Legation of both ducts resulted in a trace hyperbiliverdinemia and a distinct hyperbilirubinemia. Therefore it is suggested that either biliverdin formation occurs extrahepatically or biliverdin from the liver bile canaliculi is reduced to bilirubin in the extrahepatic biliary system in the chicken (Campbell and Coles 1986). In embryos, Vajro et al. (1995) reported that bilirubin accounted for 2.5-11.5% of total bile pigments, with the higher percentages in the early embryo. In serum, bilirubin and biliverdin were undetectable at all embryonic stages and after hatching. Elevation of serum bilirubin may occur with biliary obstruction or intravascular hemolysis. Clinical icterus is rare in psittacine birds, and

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plasma bilirubin determinations have been useful in detecting liver disease in these birds. Psittacine birds with severe liver damage or suffering from starvation frequently have a greenish discoloration to their serum and urates. This is considered to be a consequence of hyperbiliverdinemia and biliverdinuria, as biliverdin is green. Occasionally, psittacine birds with chronic liver disease will have icteric-appearing tissues, this may be caused by nonspecific reduction of bililverdin to bilirubin (Campbell and Coles 1986). In a diseased animal, serum bilirubin determinations are of value in classifying icterus and may also be used to measure the response of the liver to therapy and thus assist in making an accurate prognosis. Elevation of serum bilirubin level in the horse (25-75 mg/dl) has been reported in cardiac insufficiency, hemolytic diseases, and primary hepatic disorders (Coles, 1986a). Clearance of Foreign Dyes from The Serum: The clearance of a foreign dye from the serum following parenteral injection is a measure of both biochemical integrity and blood flow in the liver. Delay in removal of such a dye from the blood may be an indication of hepatic necrosis or fibrosis. Dye that has been widely used is BSP. When BSP is injected intravenously, it is taken up rapidly, concentrated by the liver, and excreted into the bile. BSP excretion involves the following steps:(1) transfer of the dye from blood to hepatic parenchymal cells, (2) brief storage bound to ligand and Z protein, (3) conjugation with glutathione, and (4) active excretion of conjugated dye into bile. A small fraction may be excreted in an unconjugated form. (Coles, 1986a). Use of this test in birds has been limited but it is clear that the test is applicable. The simplest method is to give a standard intravenous dose (generally 5 mg/kg body weight) and take a first plasma sample after 5 min and a second after 30 or 45 min; the quantity of BSP in the 2nd example is expressed as a percentage of that in the first sample thus giving a simple percentage retention value. The avian liver is extremely efficient in abstracting BSP from the plasma and 10 min after a standard dose of 5 mg/kg BW, the plasma BSP level is too low for accurate determination. Campbell and Coles (1986) increased the dose to 20 mg/kg BW so that the plasma concentration would be more readily measured. There is, however, the risk of exceeding the excretory maximum capacity with high doses, especially where the hepatic BSP excretion is reduced by drugs or disease.

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The BSP removal in turkeys and chickens is more rapid than in dog or man and analysis is somewhat more difficult (Clarkson and Richards 1971). In the dog, less than 5% BSP retention at 30 min has been accepted as normal. However, up to 10% retention 30 min following injection can occasionally be demonstrated in dogs having no apparent hepatic damage. The conditions capable of causing prolongation of BSP retention in dog are hepatic lipidosis, focal hepatitis, carbon tetrachloride poisoning, and infectious hepatities. Also, delayed BSP excretion in horse occurs in hepatic hemosiderosis, extensive lipidosis, and carbon tetrachloride intoxication (Coles, 1986a). Second: Tests Based on Specific Biochemical Functions Plasma Proteins: Plasma proteins represent a heterogeneous group of chemical compounds, albumin, globulins, fibrinogen, glycoprotein, and lipoproteins. Gamma globulins are synthesized by lymphoid cells of the lymph nodes, spleen, and bone marrow, whereas albumin, fibrinogen, and prothrombin are thought to be formed solely in the liver, which is also the primary site of formation of the alpha and beta globulins. Plasma gel electrophoresis can be used to accurately determine albumin concentration and globulin distribution. Electrophoresis is useful for staging acute and chronic inflammatory conditions and for monitoring therapeutic response in birds, which frequently show few overt clinical signs (Cray and Tatum, 1998). Plasma proteins identified in the classic banding pattern of avian species include transthyretin (prealbumin fraction), albumin, -1-antitrypsin (-1-globulin fraction), -2-macroglobulin (-2- lobulin fraction), fibrinogen, -lipoprotein, transferrin, complement, and vitellogenin (-lobulin fraction), and immunoglobulins and complement degradation products (-globulin fraction) (Cray and Tatum, 1998; Chang et al., 1999). Plasma total protein: Concentration of total protein may be of significance both nostically and prognostically. Any abnormality in plasma proteins indicates that some pathologic, or other induced factor, (e.g. water balance, and nutritional state), is responsible. Also, alterations in plasma protein values may be observed in association with both kidney and liver diseases (Coles, 1986a). The plasma protein concentration of avian blood is lower than that of mammals. Most normal birds have plasma total protein values between 3 and 8 gm/dl. A reading less than 3.0 gm/dl usually indicates hypoalbuminemia, because albumin is the greatest individual protein fraction in avian plasma. Total protein values less than 2.5 gm/dl indicate a grave prognosis; birds with severe hypoproteinemia rarely survive

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(Altman, 1979). Hypoproteinemia can occur with chronic renal or hepatic disease, malnutrition, malabsorption (e.g., intestinal parasitism), or chronic blood loss. Elevated total protein (hyperproteinemia) greater than 6.0 mg/dl occurs with dehydration, shock, or if there is an increase in total globulins (Altman, 1979; Campbell and Coles 1986). Plasma Albumin: Albumin is the largest protein fraction in normal avian serum. Avian albumin is similar in structure to mammalian albumin. Albumin binds and transports anions, cations, fatty acids, and thyroid hormones (Ivins et al. 1978). Seventy-five percent of the plasma T4 is attached to albumin and 50 percent of plasma T3 is associated with albumin in chickens (Butler, 1983). Therefore, hypoalbuminemia will affect the blood concentration of the albumin-transported compounds. Hypoalbuminemia are presented in starvation and malnutrition and in chronic gastrointestinal disease in which there is interference with protein digestion and absorption. Deficient synthesis of albumin occurs commonly in association with chronic hepatic disease like hepatitis and cirrhosis in the dog (Coles, 1986a). Decreased albumin concentration has been observed in birds with maldigestion, malabsorption, and protein-losing enteropathy. Other differential diagnoses for hypoalbuminemia include protein-losing nephropathy and liver failure (Stone and Redig, 1994;Wilson et al. 1999; Harr, 2002). Hyperalbuminemia is rarelly seen except in the presence of acute dehydration and shock. Increases in albumin are usually masked by increases in total plasma volume. Hyperalbuminemia has been associated with pituitary neoplasms that produce an increased level of growth hormone (Campbell and Coles 1986). Plasma Globulins: The globulin component of avian serum protein is composed of separate alpha, beta, and gamma fractions. The primary function of alpha and beta globulins is to serve as carriers. The alpha globulins include glycoproteins that has been termed ceruloplasmin, hepatoglobin which binds hemoglobin, and alph2 macroglobulin. Transcortin, an alpha globulin, is the primary transport protein for corticosterone in chicken plasma. Infections produce marked increases in ceruloplasmin, fibrinogen, and hepatoglobin. Therefore, the alpha globulins increase with infections. Alpha globulins increase with tissue destruction such as following surgery or in birds with osteomyelitis and decrease with liver disease, malabsorption, or malnutrition (Ivins et al., 1978). In domestic fowl, cholecalciferol and 25-hydroxycholecalciferol are carried by the beta globulins. The beta globulins are usually elevated when there is an increase in beta-lipoprotein in chronic infectious disease (Ivins,

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1978). Transportation of iron may be related to the beta globulins. The glycoprotein responsible for this binding of iron has been called transferrin or siderophilin (Coles, 1986a). The gamma globulins (immunoglobulins) include the circulating antibodies, which elevated with chronic inflammation. In general, an increase in concentration of gamma globulin accompanies with a rise in antibody titer. Phosvitin and other phospholipoproteins that transport iron during ovulation also migrate with gamma globulin fraction (Ivins et al., 1978). Antibodies may migrate in both beta and gamma globulin ranges, IgG migrates in gamma globulin whereas IgM migrates in beta globulin. In the domestic fowl IgA is structurally different from mammalian IgA but has a similar faction. Avian IgA is primarily found in external secretions, with only about 4 percent located in serum (Butler, 1983). Avian IgG and IgM are comparable to mammalian IgG and IgM (Campbell and Coles, 1986). Elevations occur in gamma globulins in bacterial infections, viral infections, parasitism, and liver diseases. Increases have also been associated with certain types of neoplasms, notably lymphosarcoma, and plasmacytoma (Coles, 1986a). In conclusion, the absolute fall in serum albumin concentration resulting from a disturbance of normal synthesis by the liver is not an early biochemical alteration. Such a fall is found more commonly in chronic liver diseases such as sub-acute hepatitis or diffuse fibrosis. In acute hepatitis, changes in albumin are less significant, but an elevation in gamma globulin occurs rather consistently. Albumin/globulin (A/G) ratio may be decreased in inflammation, protein-losing nephropathy, and liver failure (Jones, 1999). Females of oviparous species may have a physiological decrease in A/G ratio concurrent with an estrogen- induced hyperproteinemia composed of proteins involved in egg formation (Simkiss, 1967; Lumeij, 1997). The majority of yolk proteins and chalazae band in the globulin region and cause a marked increase in globulin fractions. Albumin concentration may be mildly increased during egg formation. Egg formation therefore results in a decreased A/G ratio that is not indicative of disease. Carbohydrate Metabolism Test Serum Glucose: Because of the ability of the liver to participate in carbohydrate metabolism and its inherent ability to metabolize increased quantities of carbohydrates, several tests have been developed for the estimation of this metabolic function. The normal blood glucose level for most birds is 200-450 mg/dl, which is much higher than for any mammalian species (Campbell and Coles 1986). Moreover, Diamond et al. (1986)

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confirmed that healthy birds maintain blood glucose concentrations of >150 mg/dl with levels up to 800 mg/dl in hummingbirds. Similarly, Clarenburg (1992) demonstrated that the normal range of plasma glucose in chicken is (130-270 mg/dl). The serum glucose values less than 70 mg/dl are a grave sign (Altman, 1979). Hypoglycemic convulsions may occur in birds of prey having glucose values less than 80 mg/dl (Wallner-Pendleton et al., 1993). Hypoglycemia in birds may occur with starvation, malnutrition (such as hypovitaminosis A), high protein diets and urea-containing diets, hepatopathies (e.g. acute hepatities, Pacheco’s parrot disease, and chronic liver disease),septicemias, and enocrinopathies. Hypovitaminosis A, high protein diets, and urea-containing diets result in hypoglycemia due to malabsorption of glucose from degenerated intestinal brush borders, malreabsorption due to degenerating renal tubules or low glucose-6phosphatase activity. Low glucose levels were attributed to flight training after restricted food intake. Hyperglycemia in birds occurs with stress, iatrogenic glucocorticoid excess, hyperthermia, and diabetes mellitus. Hazelwood (1986) postulated that hyperglycemia is induced in birds by high levels of endogenous or exogenous glucocorticoids. Geese with lead poisoning have a slight hyperglycemia. Diabetes mellitus in birds is characterized by glucose levels greater than 700 mg/dl (Driver, 1981; Harr, 2002). Lipid Metabolism Tests Serum Cholesterol and Fatty Acids: The liver is involved in many phases of lipid metabolism including synthesis, esterification, and excretion of cholesterol. Lipids in avian blood are similar in quantity and quality to those of mammals (Griminger, 1976). Circulating lipids are derived from intestinal absorption of dietary lipids, hepatic synthesis, or mobilization from fat deposits. Dietary lipids absorbed from the intestines enter the systematic circulations via the portal vein as very low density lipoprotein (VLDL), whereas mammals use a more highly developed lymphatic system and transport dietary lipid in larger particles (Butler, 1983). Plasma lipids are classified as neutral fats (triglycerides), phospholipids, cholesterol ester, free fatty acids and fat-soluble compounds such as the fat-soluble vitamins. Cholesterol metabolism in companion avian species is similar to that of mammals, but there are differences in the clinical presentation of birds with abnormal cholesterol values. In oviparous species, such as birds, a marked increase in plasma cholesterol concentration can be seen during

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vitellogenesis and egg formation (Johnson 2000). Cholesterol values may be increased before the egg(s) can be visualized on radiographs (Harr et al., 2001). Cholesterol levels in avian blood are affected by age, heredity, nutrition and various diseases. The normal serum cholesterol value for most birds is 100 to 200 mg/dl (Rivetz et al., 1977; Christie et al., 1979). Similarly, Clarenburg (1992) noted that the cholesterol value in chicken ranged between 125 and 200 mg/dl. Hypercholesterolemia has been associated with starvation, high levels of dietary fat, hypothyroidism, and liver disease (Griminger, 1976). Low serum cholesterol has been associated with bacterial septicemias and liver disease (Christie et al., 1979). A decrease in serum cholesterol and total lipids occurs in domestic fowl with Borrelia anserine infections. This occurs because of decreased intestinal absorption due to enteritis or decreased hepatogenic lipogenic activity (Rivetz et al., 1977). Free fatty acid levels in the blood of young chicks will be elevated following starvation (Langslow et al., 1970). Lead poisoning in geese causes a decrease in plasma free fatty acids (March et al., 1976). Iron deficiency anemia in chickens is accompanied by hyperlipidmia due to reduction of lipoprotein lipase activity; which is required for lipid deposition in adipose tissue. Third: Tests Based on Serum Enzyme Activity Enzyme activities vary greatly among tissues and species of birds. It is important to realize that the activity of a particular enzyme may be high in one organ or tissue, or even specific for that tissue. Alterations in serum enzyme activity due to malfuctioning of the liver occur as a result of three processes: 1. An elevation of enzyme due to disruption of hepatic cells as a result of necrosis or as a consequence of altered membrane permeability. This enzymes are GPT, GOT, arginase, glutamic dehydrogenase(GD), sorbitol dehydrogenase (SD), and lactic dehydrogenase(LDH). 2. A decrease in concentration in the serum resulting from impaired synthesis by the liver (cholin esterase). 3. An elevation in enzyme levels due to cholestasis (alkaline phosphatase,and γ glutamyl transferase). Transaminases (Amino Transferases)

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Transaminases function to catalyze the transfer of an amino group from an amino acid to a keto acid. The two clinically important amino transferases are alanine amino transferase (ALT) [formerly known as glutamic pyruvic transaminase (GPT)] and aspartate amino transferase (AST) [formerly called glutamic oxaloacetic transaminase (GOT)]. Aspartate Amino Transferase (AST) (Formerly GOT): The distribution of AST in avian tissue varies among the species. The highest AST activity in the chicken and goose occurs in heart muscle followed by liver and skeletal muscle (Bogin and Israeli 1976). The highest AST activity in tissue of the turkey is in heart muscle, followed by liver, kidney, brain and skeletal muscle. The distribution in duck tissues is skeletal muscle, heart muscle, kidney, brain and liver. AST activity in the blood of ducks is higher in erythrocytes than in plasma or serum, suggesting that hemolysis will elevate serum activity (Rozman et al., 1974). Serum AST is not liver-specific in birds; however, increased activity has been associated with hepatocellular damage in chickens, turkeys, caged birds, and ducks. The most common cause of elevated serum AST activity in caged birds is hepatic disease. Birds with serum AST activity greater than 230 IU/L are considered abnormal (Altman, 1979; Martin et al., 1983; Kaplan and Pesce 1989; Calenburg 1992). A moderate increase in serum AST activity (2 to 4 fold increase) is seen with soft tissue injury, whereas liver necrosis causes a more marked elevation (Ivins et al., 1978). Moderate increases in serum AST activity occur following intramuscular injections. Plasma AST activity returned to normal reference values within 100 hours after doxycycline-induced muscle trauma in pigeons. AST activity currently is considered to be a very sensitive but nonspecific indicator of hepatocellular disease in avian species, and is frequently used with the muscle-specific enzyme, creatine kinase (CK) to differentiate between liver and muscle damage (Dabbert and Powell,1993; Jaensch et al., 2000). Slight elevations in serum AST may be associated with glucocorticoid excess. Stress induced increased serum AST activity would be accompanied by stress leucotomy (Curtis et al., 1980). Therefore, care must be taken in interpreting the results of serum AST activity. Alanine Amino Transferase (ALT) (Formerly GPT): The ALT activity of various avian tissues varies with the species. In turkeys ALT activity is highest in skeletal muscle and low in the liver and heart. Heart, skeletal muscle, liver, and lung tissues have low ALT activity in chickens and geese (Curtis, 1980). There is little ALT activity in normal chicken plasma (Rozman, et al., 1974). Some authors report elevations in serum

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ALT activity in raptors, chickens, and ducks with hepatic insult. Others believe that ALT is not a useful diagnostic test for liver disease in birds. Intramuscular injection of doxycycline in pigeons caused plasma ALT levels to increase above reference values for more than 200 hours. Becase of the effect of injections, ALT has poor specificity for liver disease, and the clinical relevance of an increased ALT value is decreased. For this reason, ALT frequently is omitted from avian clinical chemistry panels (Lumeij, 1997). On the other side, ALT is present in large quantities in the hepatocyte cytoplasm of dog, cat and human and it is increased in serum when cellular degeneration or destruction occurs in this organ. Therefore, ALT is a specific indicator of liver damage in these species (Martin et al. 1983; Campbell and Coles 1986; Kaplan and Pesce 1989; Calenburg 1992). But, in horses, sheep, pigs and cattle their livers don’t contain a significant level of ALT (Coles, 1986a). Lactate Dehydrogenase (LDH): Lactate dehydrogenase catalyzes the readily reversible reaction involving the oxidation of lactate to pyruvate with NAD serving as coenzyme (Martin et al 1983; Calbreath, 1992). This enzyme has wide distribution in animal tissues. High concentrations of LDH are found in liver, cardiac and skeletal muscle, erythrocytes, gut and renal cortices (Martin et al 1983; Calbreath, 1992). The highest LDH activity in chickens and geese occurs in skeletal muscle followed by heart muscle, liver, and lung. In turkeys LDH activity is highest in heart muscle, followed by skeletal muscle, liver, spleen, and lung. In the duck the highest LDH activity is in the liver. The five LDH isozymes found in mammalian tissues also occur in birds but with different distribution, for example, pigeon LDH1 and LDH2 are highest in heart muscle and LDH2, 3 and 4 are highest in the liver. Serum LDH activity will increase with hemolysis in avian species. Lactate dehydrogenase (LDH) isoenzymes are found in most avian tissues such that increased plasma LDH activity has low specificity for liver disease (Lumeij and Westerhof, 1987). Currently, LDH isoenzymes are not commonly measured for the clinical evaluation of birds. Contrary to prior statements in the literature, LDH has not proven to be useful in assessing fitness in raptors (Joseph, 1999; Harr, 2002). Elevated serum LDH activity will usually indicate hepatic disease in psittacine birds, but it is not specific for liver disease. Soft tissue injury results in moderate elevations of serum LDH (Campbell and Coles, 1986).

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In human, total LDH is increased in viral or toxic hepatitis, extrahepatic biliary obstruction, acute necrosis of the liver, and cirrhosis of liver (Kaplan and Pesce 1989; Calbreath 1992). Alkaline Phosphatase (ALP): Alkaline Phosphatase catalyzes the hydrolysis of various phosphate esters, transferring the phosphate group to an acceptor molecule. ALP has a pH optimum between 9 and 10. This enzyme is widely distributed in the body, and is found in high concentrations in bone (in the osteoblasts), intestinal mucosa, renal tubule cells and liver (Martin et al 1983; Kaplan and Pesce 1989; Calbreath 1992). Serum ALP does not appear to be an important test for hepatic disease in noncarnivorous birds. However, it does appear to be associated with intestinal and bone activity. In the domestic fowl, serum ALP activity does not become elevated with severe cholestatic liver disease. Serum ALP also appears to be associated with bone activity, because increased levels occur with bone fractures, osteomyelitis, primary and secondary hyperparathyrodism, and somatic growth (Ivins et al., 1978; Campbell and Coles, 1986). Very low levels of alkaline phosphatase (ALP) activity have been found in the liver of avian species studied (Lumeij and Westerhof 1987). Normal serum ALP activity is less than 10 IU/L in noncarnivorous birds. If serum ALP activity is greater than 40 IU/L, bone involvement should be considered (Altman et al., 1975). Marked increases in plasma ALP activity appear to be specific for osteoblastic activity and bony change associated with growth, trauma, repair, osteomyelitis, neoplasia, nutritional secondary hyperparathyroidism, and egg-shell deposition (Lumeij and Westerhof 1987). Serum ALP may be a useful test for liver disease in carnivorous birds. Increases in serum ALP have been reported in raptors with severe liver disease such as herpes inclusion-body hepatitis and cholestasis. The serum ALP activity was 5 to 6 times normal with hepatic insult compared to a 2 to 3 fold increase with osteoblastic activity in raptors. In chickens, serum AP activity is reduced with magnesium and zinc deficiencies, hypovitaminosis D3, and coccidiosis (Rozman, et al., 1974). Ducks with lead poisoning have lowered serum ALP values because lead inhibits alkaline phosphatase activity (Rozman, et al., 1974). In human, cattle, sheep, and cat, serum ALP is useful in the diagnosis of hepatobiliary disease and extrahepatobiliary obstruction such as gallstones (Kaplan and Pesce 1989; Calbreath 1992).

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Gamma- Glutamyl Transferase (GGT): The enzyme GGT is a membrane-localized enzyme that plays a major role in glutathione metabolism and resorption of amino acids from the glomerular filtrate. It is also important in the absorption of amino acids from the intestinal lumen (Kaplan and Pesce 1989; Calbreath 1992). The usefulness of GGT as a diagnostic test has not been well investigated in birds. In one study, young chickens with damage to the hepatobiliary system and pancreas showed elevated serum GGT activity (Pearson et al., 1979). Elevated serum GGT values have also been seen in caged birds, primarily psittacine, with liver disease (Campbell and Coles, 1986). Gamma glutamyltransferase (GGT) is probably specific to biliary and renal epithelium in birds, similar to dogs and cats (Lumeij, 1987; Kramer and Hoffmann (1998). Although GGT is considered “insensitive and inconsistent in the diagnosis of liver disease” in birds, Lumeij (1997) found increased plasma GGT activity in the majority of pigeons with experimentally-induced liver disease. Marked increases in GGT activity in birds with bile duct carcinoma also have been reported (Phalen et al., 1997). Although reference intervals have not been established, GGT values of 0-10 U/L are considered “normal” at the Schubot Exotic Bird Health Center (College Station, Texas, USA). GGT values appear to be slightly higher in older Amazon parrots, which may have up to 16 U/L without other evidence of liver disease. These GGT values are higher than the reference intervals for GGT reported by Lumeij (1997) of < 3 or 4 U/L in most species except Amazon parrots, which had a high normal value of 10 U/L. Differences in methodologies for measuring GGT may account for differences in reference values. There are numerous reports of birds with bile duct carcinoma or cholangiocarcinoma, in which no concurrent increase in GGT activity was reported (Coleman,1991; Munson et al., 1996). In human, serum GGT is generally elevated as a result of liver disease. Serum GGT is elevated earlier than other liver enzymes in diseases such as acute cholecystitis, acute and subacute liver necrosis and neoplasms of multiple sites at which liver metastases are present (Kaplan and Pesce 1989; Calbreath 1992). In the dog, GGT might be useful in the detection of cholestasis. In a study of the effect of common bile duct ligation on GGT, it was found to cause increases in serum GGT activity (Coles, 1986a).

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Iditol Dehydrogenase (ID): Iditol dehydrogenase (formerly sorbitol dehydrogenase SDH) is a liver specific enzyme present in the liver in all animal species. Consequently serum elevations of ID occur when the liver is damaged. In ruminants serum ID activity increased in facioliasis, hepatic lipidosis, and on the conditions with associated hepatocyte damage (Coles, 1986a). In the fowl, serum ID is unstable and must be assayed immediately after sample collection. Normal domestic fowl have low serum ID activity (Campbell and Coles, 1986). KIDNEY FUNCTION TESTS The methods of study that are employed most widely and are of the greatest value fall into several categories: (1) Urine specific gravity and effect of water deprivation. (2) Estimation of non-protein nitrogen levels in the blood. (3) Tests based on the clearance concept. First: Urine Specific Gravity This determination measures the ability of the kidney to concentrate dilute urine. Specific gravity determinations are of value in detecting functional changes that occur during the course of a renal disease. If urine specific gravity is 1.030 or greater, it must be assumed that the kidneys are capable of performing their (Coles, 1986b). Fixation of specific gravity (isosthenuria) at or near that of glomerular filtration (1.008 to 1.012) is a consistent finding in chronic and acute renal disease in which a great percentage of the functional tubules have been damaged. It has been estimated that impairment of ability of the kidneys to concentrate or dilute urine is usually not detectable by means of specific gravity determinations until at least two thirds of the total functional parenchyma has been incapacitated. In the uremic patient, a specific gravity greater than 1.030 may be a favorable prognostic sign. Such a specific gravity is an indication that there are enough functional nephrons present to concentrate urine and may suggest that the uremia is prerenal in origin (Coles, 1986b). Although a specific gravity of 1.001 to 1.006 is extremely low and may indicate a lack of ADH or a kidney that is incapable of responding to ADH, it is also a reflection of functional kidneys, since kidneys must be working in order for solute to be removed from the glomerular filtrate (Coles, 1986b).

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Urine Osmolality: Determination of osmolality is another technique for

measuring the ability to concentrate urine. This is a measurement of the number of particles of solute per unit of solvent. The osmolality of a liquid is most readily determined by evaluating the free point. As the number of solute particles increases, the freezing point becomes progressively lower and can be measured utilizing an osmometer (Coles, 1986b). One osmole is that quantity of an ideal solute in 1 kg of water that have a freezing point of - 1.08 °C as compared with the freezing point of pure water. The unit for recording the osmotic concentration of urine is that milliosmole (mOsm), which is equal to 0.001osmole. The normal osmotic concentration of the body fluids ( transcellular, intracellular, and interstitial) is relatively constant at approximately 300 mOsm. Osmotic concentration of normal urine is variable and is dependent upon the electrolyte and fluid balance of the body as well as the nitrogen content of the diet. Normal values for urine osmolality have been reported to be between 860- 1920 mOsm / kg BW in the cow, 200- 2000 mOsm/ kgBW in the dog and 5001200 mOsm/ kgBW in the cat (Coles, 1986b). The relationship between urine osmolality (total solute) and apecific gravity is only approximate, as urine specific gravity is altered by abnormal solutes, such as protein and glucose. This change in specific gravity is dependent upon molecular size and weight of the solutes as well as the number of molecules of solute. As a consequence, equal numbers of molecules of albumin, globulin, fibrinogen, glucose, sodium, chloride and urea each have different quantitative affects on specific gravity (Coles, 1986b). The ratio of urine osmolality (Uosm) to plasma osmolality (Posm) may be a good index of renal function. The normal osmotic concentration of plasma is approximately 300 mOsm/ kg of water. A ratio of U/Posm greater than 1 indicates that the kidneys are capable of producing urine that is more concentrated than is plasma. A U/Posm ratio of less than 1 indicates that kidneys are capable of absorbing solute in excess of water. A U/Posm ratio of 1 indicates that water and solute are being eliminated in a state which is osmotic with plasma. Osmoregulation in birds is accomplished by contributions from the kidneys, intestinal tract, salt glands, and, to some extent, the skin and respiratory tract (Goldstein and Skadhauge, 2000). Urine can be actively retropulsed from the urodeum to the coprodeum of the cloaca and then to the rectum and potentially the large intestine, where water can be reabsorbed and electrolytes can be modified (Goldstein and Braun, 1988). This affects the specific gravity, electrolyte concentrations, and bacterial

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contamination of urine. Urine specific gravity of noncarnivorous birds is 1.002 to 1.033 and will vary depending upon the sate of hydration and osmolality. It is difficult to obtain a specific gravity reading on semisolid urine, but this can be done on watery urine using a refractometer (Campbell and Coles, 1986). Moreover, Braun (1998) confirmed that specific gravity in most clinically normal birds has been reported as 1.005-1.020, and avian urine is usually acidic. Second: Nonprotein Nitrogen Levels In Blood The term nonprotein nitrogen (NPN) is used to identify nitrogen containing components of serum or plasma that are not associated with protein. Nonprotein nitrogens include urea, creatinine, creatine. uric acid, ammonia, and amino acids. Uric Acid: Uric acid is the primary catabolic product of protein, nonprotein nitrogen, and purines in birds. The avian kidney excretes uric acid primarily by tubular excretion, unlike the mammalian system that excretes urea entirely by filtration. The clearance of uric acid by tubular secretion surpasses the glomerular filtration by a factor of 8 or higher, representing 80 to 90 percent of the total excretion (Skadhauge, 1983). The rate of uric acid excretion is largely independent of the hydration status and rate of urine flow in birds. The rate of uric acid excretion is primarily influenced by the plasma uric acid concentration and renal portal blood flow. A bird in normal nitrogen and acid-base balance will excrete approximately 80% of the total nitrogen as uric acid, 15% as ammonia, and 1 to 10 % as urea (Skadhauge, 1983; Goldstein and Skadhauge, 2000). The normal blood uric acid value for most bids is 2- 15 mg/dl. Uric acid values (greater than 20 mg/dl) are considered elevated. Hyperuricemia in birds occurs with starvation, gout (visceral and articular), massive tissue destruction, and renal disease (Rivetz et al., 1977; Ivins et al., 1978). Nephrocalcinosis due to high levels of dietary calcium or hypervitaminosis D3 will result in an elevated blood uric acid level. Plasma uric acid increases with loss of tow third of the functional renal mass in birds. Hyperuricemia due to renal disease is the result of decreased rate of tubular excretion plus the poor nutritional status, which increases uric acid production as body proteins are degraded. Hypovitaminosis A causes impaired function and is accompanied by hyperuricemia. Diets high in protein and urea will elevate serum uric acid due to an increase in uric acid biosynthesis. Excess ammonia absorption from the large intestine can elevate serum uric acid. Renal neoplasms have also been associated with

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Hyperuricemia (Altman, 1979). Serum uric acid may be art factually increased if the toenail clip method of blood collection is used and the nail is not properly cleaned of urates from the bird’s droppings (Campbell and Coles, 1986). Urea Nitrogen : Birds are uricotelic and produce uric acid as the major nitrogenous end product of metabolism, whereas mammales are ureotelic and produce primarily urea as the end product of nitrogen metabolism. Therefore, blood urea nitrogen is not a useful test of renal function in birds. Ammonia may comprise up to 25% of the total nitrogen in urine. Blood ammonia concentration has not been evaluated for diagnostic relevance in companion avian species. Acid-base alterations may result in increased ammonia concentration in birds (Long and Skadhauge, 1983). Studies of birds of prey indicate that the blood urea nitrogen level will become elevated only after major kidney damage. These birds are probably exposed to higher levels of dietary urea than are noncarnivorous birds and excrete absorbed urea through their kidneys. Poultry fed high ureacontaining diets will show increased levels of blood urea nitrogen (Campbell and Coles, 1986). In animals, the nitrogen concentration may be increased as much as 10mg/dl if the animal is on a diet high in meat. Plasma urea nitrogen levels in dogs on a dry food diet didn’t exceed 25mg/dl and the highest recorded was 43mg/dl in one dog fed canned food. Catabolic breakdown of the tissues as a consequence of fever, trauma, infecton, or toxemia may result in a moderate increase in urea nitrogen concentartion. A similar increase may be seen in association with hemorrhage into the gastrointestinal tract. Also, kidney diseases are accompanied by changes in urea nitrogen levels, and there may be destruction of a large quantity of renal parenchyma (Coles, 1986b). Creatinine: Creatinine is not a major nonprotein nitrogencomponent of avian blood .Avian urine contains very little creatinine but much more creatine. Therefore serum creatinine has questionable value in the evaluation of renal function in birds. Many investigators think that serum creatinine may become elevated in birds with renal failure, but less reliably than uric acid. The normal serum creatinine for most birds 0.5- 1.5 mg/dl. High serum creatinine values may be seen in psittacine birds fed high levels of animal protein (Campbell and Coles, 1986). Sykes (1971) showed that the tubular secretion of creatinine can be selectively inhibited by drugs which inhibit the transport of either organic bases (e.g. priscoline) or organic acid (e.g. benemid).

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Creatine: While healthy humans excrete creatinine and little if any creatine, the reverse is the case in the fowl which presumably lacks the creatine -dehydrogenase mechanism. Plasma creatine level is between 1.58 and 0.5 (mean 0.97) mg/dl in 5-week-old cockerels and between 1.83 and 0.74 (mean 1.7) mg/dl in laying hens (Harms et al., 1995). Other Blood Chemistry Determinations As the kidneys play an important role in elimination and conservation of several chemical components of blood, renal disease may alter these blood chemical values. Calcium: Avian total calcium values can be much higher under normal physiological circumstances than would be tolerated by a mammal. Dramatic increases in plasma total calcium concentration are seen in reproductive, oviparous females due to estrogen-induced transport of calcium-bound yolk proteins to the ovary. Therefore, sex- and possibly season-specific reference values are required for accurate clinical evaluation of calcium values, although few have been published. Reproductive pathology such as egg binding and egg yolk coelomitis also can result in marked total hypercalcaemia (Jones, 1999). Serum calcium levels for most normal birds are 8-18 mg/dl. Hypercalcemia occurs with hypervitaminosis D3 or as a normal physiologic occurrence in egg-laying hens. Hypocalcemiais is associated with advanced nutritional secondary hyperparathyrodism, renal failure, hypoalbuminemia and excessive fat necrosis (Campbell and Coles, 1986). Clarenburg (1992) showed that the normal level of serum calcium in chicken is 4.5 - 6.0 mEq/L and in lying hens is 8.5 - 19.5 mEq/L. Phosphorus: Serum phosphorus levels for most normal birds is 2.0 4.5 mg/dl. Elevated serum phosphorus can be associated with renal disease in which the phosphorus level can be 9.5 mg/dl or greater. Avian renal disease is often associated with a hyperuricemia and hyperphosphatemia. Hypervitaminosis D3 will increase serum phosphorus values. Low serum phosphorus levels are seen in enteric diseases when impaired intestinal absorption of phosphorus has occured. Starvation and anorexia will produce hypophosphatemia (Campbell and Coles, 1986). Moreover, Clarenburg (1992) showed that the normal level of serum phosphorus is 3 - 6 mEq/L in chicken.

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Third: Renal Clearance Tests As a part of its normal functioning, the kidney is said to clear blood of certain of its constituents. Renal clearance is defined as the volume of blood that can be cleared of a given substance by excretion of urine for 1 min. The concept of renal clearance has contributed a great deay to the present understanding of renal function in disease and health. In theory, a substance may be excreted by (1) glomerular filtration alone, (2) filtration plus tubular excretion, or (3) filtration plus tubular reabsorption. If a substance is completely filtered at glomerulus and completely reabsorbed by the tubules, its clearance value is zero (e.g. glucose). As the degree of tubular reabsorption diminishes, the substance may appear in the urine. Its clearance then increases (e.g. urea) until if there is no reabsorption of a substance, its clearance will be equivalent to the rate of glomerular filtration (e.g. inulin and creatinine). Since the kidneys cannot excrete more of a substance in given period of time than is brought to them in the blood, the maximum limit of renal clearance is determined by renal flow (Coles, 1986b; Sturkie, 1976). Endogenous Creatinine Clearance: Endogenous creatinine clearance is based on the same concept of renal clearance but doesn’t require administration of creatinine. In dogs the test is completed as follows: 1-Remove residual urine by catheterization. 2-Rinse bladder several times with sterile physiologic saline. Discard urine and saline. 3-Begine timing urine formation upon completion of rinsing procedure. 4-Collect a serum sample approximately half way through the test period. 5-At the end of timed urine collection ( usually 20 min.), collect all of the urine by catheterization and rinse the bladder with several milliliters of sterile saline. Urine and saline are mixed and the total volume is measured. 6-Analyze the serum sample and the final urine plus saline collection for the quantity of creatinine per dl. 7-Accurately determine the dog’s weight. 8-Caculate creatinine clearance suing the following formula: Ccr = UcV/ Sc X T X BW

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Where: Ccr = ml/min/kg. Uc and Sc =mg/dl of creatinine in urine and serum respectively. V= Urine volume in milliters. T= Time in minutes. BW= Body weight in kilograms. The normal Ccr for dogs is 2.8±0.96 ml/min/kg , 3.48±0.12 ml/min/kg for sheep, 1.47 ml/min/kg for the horse and 1.32 to 2.23 ml/min/kg in cattle (Coles, 1986b). In interpreting the endogenous clearance one must remember that creatinine clearance is influenced by prerenal as well as renal factors. This necessitates a careful consideration of prerenal reduction in glomerular filtration such as that associated with dehydration, cardiac insufficiency, or other factors that would reduce renal blood flow (Coles, 1986b). Using Creatinine Clearance to Measure Mineral Balance: Creatinine, a breakdown product of phosphocreatine in muscles, enters the bloodstream at the same rate at which the kidneys remove it. so no infusion is needed to maintain constant CB ( Compound X in blood). Creatinine is cleared almost entirely by glomerular filtration and at a fairly constant rate. The constancy of creatinine clearance is used as an internal standard against which the clearances of other plasma constituents can be tested. The disadvantage of determining the clearance of a single substance is that blood and urine samples must be collected over a specified period of time and the volumes recorded. An acceptable alternative to this problem is to compute a ratio of the clearance of a substance to the clearance of creatinine, using a single collection of serum and urine : Percent clearance ratio = creat. s/ceart. u XCu / CsX100 Where: CuandCs = the concentration of compound χ in urine and serum (mg/dl) respectively. creat. s and ceart. u = the concentration of creatinine in urine and serum ( mg/dl ) respectively. Using creatinine clearance as an internal standard cancels the impractical item (urine volume). Consequently, timed volumetric urine

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collections are not required. By determining the percent clearance ratio of a substance relative to creatinine, the actual loss of the substance from the body through the kidneys, can be more accurately evaluated as variability caused by evaporation of the urine sample or by differences in water balance in the patiant that cancel out ( Clarenburg, 1992). It has already been pointed out that serum concentration of minerals ( e.g. Ca++ and K+) are homeostatically controlled and ,thus, are unreliable as indicators of mineral balance. Therefore, the percent clearance ratio, relative to creatinine, is a convenient diagnostic tool to aid the clinician (Clarenburg, 1992). Sturkie (1972) demonstrated that the amount formed of creatinine in birds is negligible in relation to the amount of creatinine. Clearance studies on birds ,man, apes and certain fishes indicate that the tubules of these species secrete creatinine. The clearance of endogenous creatinine averaged 2.25 ml/min at normal or endogenous plasma levels (0.2 - 0.5 %), therefore, creatinine is not secreted but is reabsorbed. The ability of the kidney to clear the plasma of creatinine is therefore considerably lower than it is for uric acid, PTH and phenol Red (Sturkie, 1972). Endogenous Inulin Clearance: Inulin is neither reabsorbed nor excreted by the kidney tubules of mammals, amphibians, or birds and varies directly with the concentartion of inulin in the plasma. C = UxV/Px Where: C =clearance of a given substance χ in the plasma. Uχ =concentration of χ in each ml of urine. V =rate of urine formation (ml/min) Pχ = concentration of χ in each ml of plasma. The substance to be tested is usually infused at a constant rate, and urine samples are collected at regular and frequent intervals (2-7 min in chicken). Blood samples are usually taken near the middle of the collection periods. Figures for inulin clearance on a number of avian species are shown in table (1). An inulin clearance of 1.8 ml/kg/min means that for a chicken weighing 2kg, the fluid filtered through the kidneys in 1 min amounts to 3.6 ml and that in 1 hr to 216 ml or 5.18 liters in 14 hr (Sturkie, 1976).

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Endogenous Urea Clearance: Urea is the end product of purine metabolism in birds, yet it is handled by the avian kidney. It is completely filterable but it is partially reabsorbed by the kidney tubules, independently of the plasma concentration. The average urea clearance in the chicken is 1.5ml/kg/min. During hydration almost all of the filtered urea is excreted, and during dehydration nearly all (99%) is resorbed (Sturkie, 1976). Endogenous Uric Acid Clearance: Uric acid is synthesized in the liver and it is filtered by glomeruli and secreted by the kidney tubules. It is highly concentrated in the urine of birds and constitutes from 60 to 80% of the total nitrogen. Studies of uric acid clearance showed that 87-93% of the chicken’s uric acid is excreted by the tubules (Sturkie, 1976). The absolute clearance of uric at plasma levels of 6-9 mg% is approximately 30 ml/kg/min. As the plasma levels of uric acid increases the amount filtered continues trends to increase, however, at very high plasma levels the ability of the tubules to secrete uric acid declines. Clearance of uric acid in doves is about 25 ml/kg/min and of about of the same magnitude in ducks (Sturkie, 1976). Impaired renal clearance of uric acid was associated with a high level of uric acid in the blood. As protein level increased in the diet the plasma level of uric acid increased to as high as 55 mg/dl and then the tubular secretion rate of high-uric acid level was only 40% of that of the normal level (Sturkie, 1976). ANTIOXIDATIVE ENZYMES AND LIPID PEROXIDATION Antioxidative Enzymes Reactive Oxygen Species (ROS) are produced during normal cellular function. ROS include hydroxyl radicals (.OH), superoxide anion (O.2-), hydrogen peroxide (H2O2) and nitric oxide (NO). They are very transient species due to their high chemical reactivity that leads to lipid peroxidation and oxidation of some enzymes, and a massive protein oxidation and degradation (Mate´s et al.,1999). The role of oxygen derived species in causing cell injury or death is increasingly recognized: superoxide and hydroxil radicals are involved in a large number of degenerative changes, often associated with an increase in peroxidative processes and linked to low antioxidant concentration (Tamagno et al., 1998). The prevention of lipid peroxidation is an essential process in all the aerobic organisms, as lipid peroxidation products can cause DNA damage

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There are cellular defense mechanisms which are responsible for converting ROS into less harmful or harmless compounds, formed within the cells. These enzymes are antioxidative enzymes which are involving in this mechanism. These enzymes play the key roles in the detoxification processes of extremely reactive free radicals such as hydrogen peroxide (H2O2), superoxide radicals (.O2-), singlet oxygen (1O2-) and hydroxyl radical (-OH), formed within the cell. This mechanism protects the cell membrane which is sensitive to peroxidation because of polyunsaturated fatty acids content (Gokhan et al 2004). The liver and kidney are major metabolic centers inside the body, according to there multifunction and the significant role of liver in lipid metabolism they are a good target for ROS attacks, measuring such parameters may be useful in evaluating the whole body defense system. Superoxide dismutase (SOD): (EC 1.15.1.1) destroys the free radical superoxide by converting it to peroxide that can in turn be destroyed by catalase or GPX reactions. Superoxide reduces Fe(III) to Fe(II), releasing the iron from storage sites so that it can react with hydrogen peroxide and produce hydroxyl radicals. SOD converts superoxide to hydrogen peroxide and molecular oxygen.

Another function of superoxide dismutase is to protect dehydratases (dihydroxy acid dehydratase, aconitase, 6-phosphogluconate dehydratase and fumarases A and B) against inactivation by the free radical superoxide (Benov and Fridovich,1998). Four classes of SOD have been identified, containing either a dinuclear Cu, Zn or mononuclear Fe, Mn or Ni cofactors (Whittaker and Whittaker,1998). Fe-SODs and Mn-SODs show homology and posses identical metal chelating residues at the active site, sharing substantial sequence and three dimensional structural homology, while the other superoxide dismutases are structurally unrelated. The other three forms of SOD are: cytosolic Cu, Zn-SOD, mitochondrial Mn-SOD, and extracellular-SOD (EC-SOD) (Majima et al., 1998, Mate´s, 2000) Catalase (EC 1.11.1.6) is a tetrameric haeminenzyme consisting of four identical tetrahedrally arranged subunits of 60 kDa. Therefore, it contains four ferriprotoporphyrin groups per molecule, and its molecular mass is about 240 kDa. Catalase is one of the most efficient enzymes known. It is so efficient that it cannot be saturated by H2O2 at any concentration (Lledı´as et al., 1998). Catalase reacts with H2O2 to form

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water and molecular oxygen; and with H donors (methanol, ethanol, formic acid, phenol…) using 1 mole of peroxide in a kind of peroxidase activity:

H2O2 is enzymically catabolized in aerobic organism by catalase and several peroxidases. In animals, catalase and GPX detoxify H2O2. Catalase protects cells from hydrogen peroxide generated within them. Even though catalase is not essential for some cells type under normal conditions, it plays an important role in the acquisition of tolerance to oxidative stress in the adaptive response of cells (Hunt et al., 1998). The increased sensitivity of transfected enriched catalase cells to adriamycin, bleomycin and paraquat is attributed to the ability of catalase in cells to prevent the drug-induced consumption of O2. Thus, capturing H2O2 before it can escape the cell and converting it to O2. In this way, catalase can maintain the concentration of O2 either for repeated rounds of chemical reduction or for direct interaction with the toxin (Speranza et al., 1993). Glutathione peroxidase (GPX): (EC 1.11.1.19) the more important example for selenium-containing peroxidases. They catalyze the reduction of a variety of hydroperoxides (ROOH and H2O2) using GSH, thereby protecting cells against oxidative damage.

They are present in almost every cell of animals, but the tissue distribution of the isoforms shows high variation. There are several factors abrogating the activity of the enzyme. Some of these are internal, individual factors, resulting in significant variation in the enzyme activity of different organs, age groups and sex. Endocrine regulation can also control enzyme activity. However, environmental factors have also definite effect on enzyme action. Nutrition is one of the most essential factors as fat content and fatty acid composition of feed, or trace element intake as well as vitamin status of the animal play crucial role in normal enzyme activity (Mézes, et al., 2003).

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There are at least five GPX isoenzymes found in mammals. Although their expression is ubiquitous, the levels of each isoform vary depending on the tissue type. Cytosolic and mitochondrial glutathione peroxidase (cGPX or GPX1) reduces fatty acid hydroperoxides and H2O2 at the expense of glutathione. GPX1 and the phospholipids hydroperoxide glutathione peroxidase GPX4 (or PHGPX) are found in most tissues. GPX4 is located in both the cytosol and the membrane fraction. PHGPX can directly reduce the phospholipids hydroperoxides, fatty acid hydroperoxides, and cholesterol hydroperoxides that are produced in peroxidized membranes and oxidized lipoproteins (Imai et al., 1998). GPX1 is predominantly present in erythrocytes, kidney, and liver, and GPX4 is highly expressed in renal epithelial cells and testes. Cytosolic GPX2 (or GPX-G1) and extracellular GPX3 (or GPX-P) are poorly detected in most tissues except for the gastrointestinal tract and kidney, respectively. Recently, a new member, GPX5, expressed specifically in mouse epididymis, is interestingly selenium-independent (De Haan et al., 1998). GPX1 (80 kDa) contains one selenocysteine (Sec) residue in each of the four identical subunits, which is essential for enzyme activity (Ding et al., 1998). Although GPX shares the substrate, H2O2, with catalase, it alone can react effectively with lipid and other organic hydroperoxides. The glutathione redox cycle is a major source of protection against low levels of oxidant stress, whereas catalase becomes more significant in protecting against severe oxidant stress (Yan and Harding, 1997). Glucose-6- phosphate dehydrogenase (G6PD): (EC 1.1.1.49) is the key enzyme, catalyzing the first step of pentose phosphate metabolic pathway. The pentose phosphate metabolic pathway is a unique source of NADPH. (Faix, et al., 2003). G6PD is important for its role in the regeneration of the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) and the production of ribose. During cellular oxidative stress, NADPH is critical for maintaining glutathione (GSH) in its reduced form, which is essential for detoxification of reactive free radicals and lipid hydroperoxides. Another important role for NADPH is the maintenance of the catalytic activity of catalase hence, NADPH also is important for its role in the detoxification of hydrogen peroxide (Christopher, et al 2000).

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Lipid peroxidation Among the cellular molecules, lipids that contain unsaturated fatty acids with more than one double bond are particularly susceptible to action of free radicals. The resulting reaction, known as lipid peroxidation, disrupts biological membranes and is thereby highly deleterious to their structure and function. Lipid peroxidation is being studied extensively in relation to disease, modulation by antioxidants and other contexts. A large number of by-products are formed during this process. These can be measured by different assays. The most common method used is the estimation of aldehydic products by their ability to react with thiobarbituric acid (TBA) that yield ‘thiobarbituric acid reactive substances’ (TBARS), which can be easily measured by spectrophotometry (absorbs light at 532 nm and fluoresces at 553 nm). Though this assay is sensitive and widely used (Devasagayam, et al., 2003 and Eid, et al., 2003).

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REFERENCES Altman, R. B., 1979. Avian clinical pathology, radiology, parasitic and infectious disease. Proceedings of theAmerican Animal Hospital Association, A. A. H. A., South Bend. IN Altman, R. R. Montali, G. Kollias and G. J. Harrison, 1975. Avian clinical pathology evaluation panel. Annual Proceedings of the American Association of Zoo Vetrinarians. Bakalli I. B., G. M. Pesti, W.L. Ragland and V. Konjufca, 1995. Dietary copper in excess of nutrition requirement reduces plasma and breast muscle cholesterol of chicken. Poultry Sci. 74: 360 – 365. Basudde. C. D., 1982. The effect of solanum malacoxylon on serum enzyme activities, blood glucose, and cholesterol levels in chicks. Poultry Sci. 61: 1001–1002. Benov, L., Fridovich, I., 1998. Growth in iron-enriched medium partially compensates E. coli for the lack of Mn and Fe SOD. J. Biol. Chem. 273: 10313–10316. Bogin, E. and B. Israeli, 1976. Enzyme profile of heart and sketletal muscle, liver and lung of roosters and geese. Zbl. Vet. Med. A. 23: 152. Braun, E. J., 1998. Comparative renal function in birds, reptiles, and mammals. Sem Avian Exotic Pet Med.;7:62-71. Butler, E. J., 1983. Plasma proteins. In: Physiology and Biochemistry of the Domestic Fowl, edited by Freeman, B. M., Academic Press. London. Calbreath, D. F., 1992. Hepatic enzymes in health and disease. In: Clinical Chemistry, A Fundamental Txetbook. Eds., Donald F. Calbreath, W.B. Saunders Company Pp: 182 – 233. Campbell T. W., and E. H. Coles, 1986. Avian clinical pathology. In: Veterinary Clinical Pathology. Edited by Coles, E. H. pp: 279 – 301. Chang, L., S. L. Munro, S. J. Richardson and G. Schreiber, 1999. Evolution of thyroid hormone binding by transthyretins in birds and mammals. Eur J Biochem.259:534-542. Christie, G. and W. G. Halliday, 1979. Haematological and biochemical aspects of an E. Coli septicemia in Brown Leghorn chickens. Avian Pathol. 8: 45.

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Clarenburg. R., 1992. Physiological Chemistry of Animals. Pp: 127. Clarkson M. J. and T. G. Richards, 1971. The liver with special reference to bile formation, In Physiology and Biochemistry of the Domestic Fowl, edited by Bell D. J. and B. M. Freeman, Academic press, London. pp: 1104 – 1111. Coleman, C. W., 1991. Bile duct carcinoma and cloacal prolapse in an orange-winged Amazon parrot (Amazona amazonica). J Assoc Avian Vet.5:87-89. Coles E. H., 1986a. Liver function. In: Veterinary Clinical Pathology. Edited by Coles, E. H. 3rd ed. WB Saunders, pp: 129–151. Coles E. H., 1986b. Kidney function. In: Veterinary Clinical Pathology. Edited by Coles, E. H. 3rd ed. WB Saunders, pp: 189-203. Cornelius, C. E., 1981. Hepatic bilirubin 1X-alpha-glycosyltransferase activities in animals excreting primarily biliverdin into bile. Vet. Clin. Pathol. 10: 27. Cray, C. and L. Tatum, 1998. Applications of protein electrophoresis in avian diagnostics. J. Avian. Med. Surg. 12: 4-10. Curtis, M. J., H. G. Jenkins and E. J. Butler, 1980. The effect of Escherichia coli endotoxins and adrenocortical hormones on plasma enzyme activities in the domestic fowl. Res. Vet. Sci. 28: 44. Dabbert, C. B. and K. C. Powell, 1993. Serum enzymes as indicators of capture myopathy in mallards (Anas platyrhynchos). J Wildlife Dis.29: 304-309. De Haan, J., Bladier, C., Griffiths, P., Kelner, M., O’Shea, R.P., Cheung, N.S., Bronson, R.T., Silvestro, M.J., Wild, S., Zheng, S.S., Beart, P.M., Herzog, P.J., Kola, I., 1998. Mice with a homozygous null mutation for the most abundant glutathione peroxidase. GPX1, show increased susceptibility to the oxidative stress-inducing agents paraquat and hydrogen peroxide. J. Biol. Chem. 273: 22528–22536. Devasagayam T., Boloor K.and Ramasarma T., 2003. Methods for estimating lipid peroxidation: An analysis of merits and demerits. Indian J. Biochemistry & Biophysics. 40 : 300-308.

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Diamond, J. M., W. H. Karasov, D. Phan and F. L. Carpenter, 1986. Digestive physiology is a determinant of foraging bout frequency in hummingbirds. Nature.320: 62-63. Ding, L., Liu, Z., Zhu, Z., Luo, G., Zhao, D., Ni, J, 1998. Biochemical characterization of selenium-containing catalytic antibody as a cytosolic glutathione peroxidase mimic. Biochem. J. 332: 251–255. Driver, E. A., 1981. Hematological and blood chemical values of Mallard, Anasp. Platyrhynchos, darkes before, during and after remige molt. J. Wildlife Des. 17: 423. Eid, Y. Z., A. Ohtsuka and K. Hayashi, 2003. Tea polyphenols reduce glucocorticoid-induced growth inhibition and oxidative stress in broiler chickens. Br. Poultry. Sci. 44(1):127-32. Eraslan G., Akdogan M., Yarsan E., Essiz D., Sahindokuyucu Fatma, Hismiogullari S. E. and Altintas L., 2004. Effects of aflatoxin and sodium bentonite administered in feed alone or combined on lipid peroxidation in the liver and kidneys of broilers. Bull. Vet. Inst. Pulawy. 48: 301-304. Faix, Z. Faixova, E. Michnova, Varady J., 2003. Effect of Per Os Administration of Mercuric Chloride on Peroxidation Processes in Japanese Quail. Acta Vet. Brno.72: 23–26. Goldstein D. L. and E. J. Braun, 1988. Contributions of the kidneys and intestines to water conservation, and plasma levels of antidiuretic hormone, during dehydration in house sparrows (Passer domesticus). J Comp Physiol. 158B: 353-361. Goldstein, D. L., and E. Skadhauge, 2000. Renal and extrarenal regulation of body fluid composition. In: Whittow GC, ed. Sturkie’s Avian Physiology. San Diego, Calif: Academic Press; pp: 265-298. Griminger, P., 1976. Lipid metabolism. In: Avian Physiology, edited by Sturkie, P. D., Springer-Verlag, New York. Harms R. H., G.B. Russell, F. Robbins and J. Cerda, 1995. Correlation of plasma calcium with experimentally elevated cholesterol and triglycerides in laying hens. Poultry Sci. 74: 1708 –1711. Harr, K. E., 2002. Clinical Chemistry of Companion Avian Species: A Review. Vet. Clin. Pathol. 31:140-151

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Harr, K. E., A. R. Alleman, P. M. Dennis, L. K. Maxwell, B. A. Lock and R. A. Bennett, 2001. Morphologic and cytochemical characteristics of blood cells and hematologic and plasma biochemical reference ranges in green iguanas. J Am Vet Med Assoc. 218: 915-921. Hazelwood, R. L., 1986. Carbohydrate metabolism. In: Sturkie PD, ed. Avian Physiology. San Diego, Calif: Academic Press; pp: 303-325. Hornbuckle, W. E. and B. C., 1997. Tennant. Gastrointestinal function. In: Kaneko JJ, Harvey JW, Bruss ML, eds. Clinical Biochemistry of Domestic Animals. San Diego, Calif: Academic Press; pp:367- 406. Hunt, C., Sim, J.E., Sullivan, S.J., Featherstone, T., Golden, W., KappHerr, C.V., Hock, R.A., Gomez, R.A., Parsian, A.J., Spitz, D.R., 1998. Genomic instability and catalase gene amplification induced by chronic exposure to oxidative stress. Cancer Res. 58: 3986–3992. Imai, H., Narashima, K., Arai, M., Sakamoto, H., Chiba, N., Nakagawa, Y., 1998. Suppression of leukotriene formation in RBL-2H3 cells that overexpressed phospholipid hydroperoxide glutathione peroxidase. J. Biol. Chem. 273:, 1990–1997. Iqbal M., D. Cawthon, K. Beers, R. F. Wideman, and W. G. Bottje, 2002. Antioxidant enzyme activities and mitochondrial fatty acids in pulmonary hypertension syndrome (phs) in broilers, Poultry Science 81:252–260 Ivins, G. K., G. D. Weddle and W. H. Halliwell, 1978. Hematology and serum chemistries in birds of pery. IN: Zoo and wild Animal Medicine, edited by Fowler, M. E., and W. B. Saunders, Philadelphia. Jaensch, M. J., Cullen L. and S. R. Raidal, 2000. Assessment of liver function in galahs/cockatoos (Eolophus roseicapillus) after partial hepatectomy: a comparison of plasma enzyme concentrations, serum bile acid levels, and galactose clearance tests. J Avian Med Surg.14:164-171. Johnson, A. L., 2000. Reproduction in the female. In: Whittow GC, ed. Sturkie’s Avian Physiology. San Diego, Calif: Academic Press; pp:569-596. Jones, M. P., 1999. Avian clinical pathology. Vet. Clin. North. Am. Exotic. Anim. Pract.2:663-87.

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Joseph, V. 1999. Raptor hematology and chemistry evaluation. Vet. Clin. North Am. Exotic. Anim. Pract.2:689-699. Kaplan L. A. and A. J. Pesce, 1989. Liver function testes. In: Clinical Chemistry, Theory, Analysis and correlation. 3rd ed. Lawrence A. Kaplan and Amadeo J. Pesce. St. Louis, MO: Mosby, pp: 368 – 371. Kramer, J. W. and W. E. Hoffmann, 1998 Clinical enzymology. In: Kaneko JJ, Harvey J, Bruss ML, eds. Clinical Biochemistry of Domestic Animals. San Diego, Calif: Academic Press; pp: 303-325. Kubena. L. E.,R. B. Harvey, W. E. Huff, and D.E. Correir, 1990a. Efficacy of a hydrated sodium calcium aluminosilicate to reduce the toxicity of aflatoxin and T.2 Toxin. Poultry Sci. 69: 1078–1086. Kubena. L. E., R. B. Harvey, T.D. Philips, D.E. Correir and W. E. Huff, 1990b. Diminution of aflatoxicosis growing chickens by the dietary addition of a hydrated, sodium calcium aluminosilicate. Poultry Sci. 69: 727 – 735. Kubena. L. E., W. E. Huff, R. B. Harvey, A. G. Yersin, M. H. Elissalde, D. A. Wttzel, , D.E. Correir,T.D. Philips, and H. D. Petersen 1991. Effect of a hydrated sodium calcium aluminosilicate on growing turkey poults during aflatoxicosis. Poultry Sci. 70: 1823 – 1830. Langslow, D. R., E. J. Butler, C. N. Hales and A. W. Pearson, 1970. The response of plasma insulin, glucose and non-esterified fatty acids to various hormones, nutreients and drugs in the domestic fowl. J. Endocrinol., 46: 243. Latour. A. M., S. A. Laiche, J. R. Thompson, A. L. Pond, and E. D. Peebles, 1996. Continuous infusion of adrenocorticotropin elevates circulating lipoprotein cholesterol and corticosterone concentrations in chickens. Poultry Sci. 75: 1428 – 1432. Lin, G. L., J. A. Himes and C. E. Cornelius, 1974. Bilirubin and biliverdin excretion by the chicken. Am. J. Physiol. 226: 881. Lledı´as, F., Rangel, P., Hansberg, W., 1998. Oxidation of catalase by singlet oxygen. J. Biol. Chem. 273: 10630–10637. Long, S., E. Skadhauge, 1983. Renal acid excretion in the domestic fowl. J Exp Biol.104: 51-58.

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Lumeij, J. T., 1987. A contribution to clinical investigative methods for birds, with special reference to the racing pigeon, Columba livia domestica. Utrecht: Rijksuniversiteit Utrecht. Lumeij, J. T., 1997. Avian clinical biochemistry. In: Kaneko JJ, Harvey JW, Bruss ML, eds. Clinical Biochemistry of Domestic Animals. San Diego, Calif: Academic Press; pp: 857-884. Lumeij, J.T. and I. Westerhof, 1987. Blood chemistry for the diagnosis of hepatobiliary disease in birds. A review. Vet Q.;9:255-261. Majima, H., Oberley, T.D., Furukawa, K., Mattson, M.P., Yen, H.C., Szweda, L.I., St. Clair, D.K., 1998. Prevention of mitochondrial injury by Mn-SOD reveals a primary mechanism for alkalineinduced cell death. J. Biol. Chem. 273: 8217–8224. March, G. L., T. M. Holn, B. A. Mckeon, L. Sileo and J. G. George, 1976. The effects of lead poisoning on various plasma constituents in the Canada goose. J. Wildlife Des.12: 14. Martin, D. W., P. A. Mayes, and V. W. Rodwell (1983). General properties of enzymes. In: Harper’s Review of Biochemistry. Lange Medical Publications. Los Altos, California, pp: 60 – 61. Mate´s J. M., 2000. Effects of antioxidant enzymes in the molecular control of reactive oxygen species toxicology. Toxicology 153: 83–104. Mate´s, J.M., Pe´rez-Go´mez, C., Nu´n˜ez de Castro, I., 1999. Antioxidant enzymes and human diseases. Clin. Biochem. 3: 595– 603. McDaniel L. S. and H. A. Dempsey, 1961. The effect of fasting upon plasma enzyme levels in chickens. Poultry Sci. 41: 994 – 998. Mézes M., Erdélyi M., Shaaban G., Vir G., Balogh K., and Wéber M., 2003. Genetics of glutathione peroxidase. Acta Biol Szeged 47:135138. Munson, L., V. L. Clyde and S. E. Orosz, 1996. Severe hepatic fibrosis and bile duct hyperplasia in four Amazon parrots. J Avian Med Surg. 10: 252-257. Nicol C. J., Zielenski J., Tsui L-C, and Wells P G., 2000. An embryoprotective role for glucose-6-phosphate dehydrogenase in developmental oxidative stress and chemical teratogenesis. FASEB J. 14: 111–127.

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Pearson, A. W., E. J. Butler and G. R. Fenwick, 1979. Rapeseed meal and liver damage: Effect on plasma enzyme activities in chicks. Vet. Rec. 105: 200. Phalen, D. N., L. Homco and L. Jaeger, 1997. Investigations into the etiologic agent of internal papillomatosis of parrots and ultrasonographic and serum chemical changes in Amazon parrots with bile duct carcinomas. Proc Assoc Avian Vet. Lake Worth, Fla: Assoc Avian Vet Publications; pp: 53-56. Rivetz, B., E. Bogin, Y. Weisman, J. Avidar and A. Hadani, 1977. Changes in the biochemical composition of blood in chicken infected with Borrelia anserins. Avian Pathol., 6: 343. Roland. D. A. , D. N. Marple, and R. N. Brewer, 1983. Serum progesterone, enzymes, and electrolytes of hens laying a low or high incidence of shell-less eggs. . Poultry Sci. 62: 917 – 922. Rozman, R. S. L. N. Locke and S. F. McClure, 1974. Enzyme changes in mallard ducks fed iron or lead shot. Avian Dis. 18: 435. Simkiss K., 1967. Calcium in Reproductive Physiology: A Comparative Study of Vertebrates. New York, NY: Reinhold Publishing Corp, pp: 155-197. Skadhauge, E., 1983. Formation and composition of urine. In: Physiology and Biochemistry of the Domestic Fowl, edited by Freeman, B. M., Accademic Press, London. Speranza, M., Bagley, A.C., Lynch, R.E., 1993. Cells enriched for catalase are sensitized to the toxicities of bleomycin, adriamycin, and paraquat. J. Biol. Chem. 268:19039– 19043. Stone, E. G. and P. T. Redig, 1994. Preliminary evaluation of hetastarch for the management of hypoproteinemia and hypovolemia. Proc Assoc Avian Vet. Lake Worth, Fla: Assoc Avian Vet Publications; pp: 197-199. Sturkie P. D., 1976. Secretion of gastric and pancreatic juice, pH of tract, digestion in alimentary canal, liver and bile, and absorption. In: Avian Physiology edited by Sturkie P. D. New York, SpringerVerlag. Sykes A. H., 1971. Formation and composition of urine, In: In Physiology and Biochemistry of the Domestic Fowl, edited by Bell D. J. and B. M. Freeman, Academic press, London.

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Tamagno, E., Aragno, M., Boccuzzi, G., Gallo, M., Parola, S., Fubini, B., Poli, G., Danni, O., 1998. Oxygen free radical scavenger properties of dehydroepiandrosterone. Cell Biochem. Funct. 1: 57– 63. Vajro,

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3rd International Poultry Conference 4 – 7 Apr. 2005 Hurghada - Egypt Table (1) Figers for inulin clearance in Aves. Species Mean inulin clearance (ml/kg/min) 1.7-1.84 Chicken

Duck

N2

Condition

Hydration

1.37

No water given

2.45

Bird very hydrated

3.00

Laying hens not fasted

1.96 2.1

Not stated Sea water

2.5

Fresh water

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3rd International Poultry Conference 4 – 7 Apr. 2005 Hurghada - Egypt

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Reference values of some chemical constituents and enzymes in avian species. Constituents

Range

Species

Authors

Total Protein (gm/dl)

3.0- 8.0 6.1-6.4 2.95-2.99 2.54 2.68

Chicken Chicken Turkey Chicken Chicken

Campbell and Coles, 1986 Waldroup et.al, 1965 Kubena et al., 1991 Kubena, 1990a Kubena, 1990b

Albumin (gm/dl)

1.10-1.21 1.2 1.66-1.97 1.45-2.69 2.7-4.6 1.7-2.2 l 1.3-2.2

Turkey Chicken Chicken Guinea Fowl Rabbit Duck (Anas sp.) Pigeon

Kubena et al,1991 Kubena et al.,1990b Sturkie, 1976 Sturkie, 1976 Okerman, 1994 Harr, 2002 Harr, 2002

Globulin (gm/dl)

2.33-3.27 1.35- 1.98 1.5- 2.8 3.5-6.0

Chicken Guinea Fowl Rabbit Duck (Anas sp.)

Sturkie, 1976 Sturkie, 1976 Okerman, 1994 Harr, 2002

Glucose

200-450 mg/dl 130-270 mg/dl 305-312 mg/dl 13.95 mmole/L 75-150 mg/dl 127-319 mg/dl 232-369 mg/dl

Chicken Chicken Turkey Chicken Rabbit Duck (Anas sp.) Pigeon

Campbell and Coles, 1986 Clarenburg, 1992 Kubena et al., 1991 Basudde, 1982 Okerman, 1994 Harr, 2002 Harr, 2002

Cholesterol (mg/dl)

100-200 125-200 132.1-154.4 96.0 150.0 113-155 169.0 133.0 104-244

Chicken Chicken Chicken Chicken Chicken Turkey Chicken Chicken Duck (Anas sp.)

Campbell and Coles, 1986 Clarenburg, 1992 Bakalli et al., 1995 Harms et al., 1995 Latour et al., 1996 Kubena et al., 1991 Kubena et al., 1990a Kubena et al., 1990b Harr, 2002

AST (GOT) (IU/L)

230 112-122 104-124

Chicken Chicken Chicken

35-60 12-73 45-123

Rabbit Duck (Anas sp.) Pigeon

Campbell and Coles, 1986 Walzeml et al., 1993 McDaniel and Dempsey, 1962 Okerman, 1994 Harr, 2002 Harr, 2002

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3rd International Poultry Conference 4 – 7 Apr. 2005 Hurghada - Egypt

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Continue: Reference values of some chemical constituents and enzymes in avian species. Constituents

Range

Species

Authors

ALT (GPT) (IU/L)

1.63 7.47 20-50

Chicken Chicken Rabbit

Basudde, 1982 Roland et al., 1983 Okerman, 1994

LDH (IU/L)

393-442 568 452-1777 94.3-104.7 120-246 30-205

Turkey Chicken Chicken Rabbit Duck (Anas sp.) Pigeon

Kubena et al., 1991 Roland et al., 1990b McDaniel and Dempsey, 1962 Okerman, 1994 Harr, 2002 Harr, 2002

ALP(IU/L)

1343-1409

Turkey

Kubena et al., 1991

5332

Chicken

Kubena et al., 1990b

186

Chicken

Roland, 1983

552.12

Chicken

Basudde, 1982

59-223

Chicken

McDaniel and Dempsey, 1962

18-192

Rabbit

Okerman, 1994

GGT(IU/L)

80.6-32.8 9.0 12.69 0.44 0.0-7

Chicken Chicken Chicken Turkey Rabbit

Walzeml et al., 1993 Kubena et al., 1990b Kubena et al., 1990a Kubena et al., 1991 Okerman, 1994

Creatinine (gm/dl)

1.0-2.0 0.5-1.5 0.9 7.0-15 0.26-0.40

Chicken Most birds Chicken Rabbit pigeon

Clarenburg, 1992 Campbell and Coles, 1986 Abdelaziz et al., 1995 Okerman, 1994 Harr, 2002

Uric Acid (gm/dl)

1.0-2.0 1.0-7.0 1.25 6.2 2.0-15 2.0-12.0 2.5-12.6

Chicken Laying hens Turkey Chicken Most birds Duck (Anas sp.) pigeon

Clarenburg, 1992 Clarenburg, 1992 Kubena et al., 1991 Kubena, 1990b Campbell and Coles, 1986 Harr, 2002 Harr, 2002

Urea(gm/dl)

0.4 – 1.0 12.42

Chicken Chicken

Clarenburg, 1992 Abdelaziz et al., 1995

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3rd International Poultry Conference 4 – 7 Apr. 2005 Hurghada - Egypt

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Continue: Reference values of some chemical constituents and enzymes in avian species. Constituents

Range

Species

Authors

Calcium (mEq/L)

4.5 – 6.0 8.5 – 19.5 8.7-12.7 7.6-10.4

Chicken Laying hens Duck (Anas sp.) pigeon

Clarenburg, 1992 Clarenburg, 1992 Harr, 2002 Harr,2002

Phosphorus (mEq/L)

3.0 – 6.0 4.0-6.2

Chicken Rabbit

Clarenburg, 1992 Okerman, 1994

SOD

1361 ± 48.37 ( U.g-1 Hb) 7.41 ± 1.18 (U/ mg protein) 6.53 ± 1.04 (U/ mg protein) 1.26 (U/ mg protein) 0.31 ± 0.40 (U/ mg protein)

Japanese Quail (liver) Broiler (liver) Broiler (kidney) Broiler (liver) Broiler (liver)

Faix, et. al., 2003 Eraslan, et. al., 2004 Eraslan, et. al., 2004 Iqbal, et. al., 2002 Young, et. al., 2003

Catalase

64.23 ± 4.86 (µmoles H2O2 decomposed/ min/mg protein ) 23.69 ± 4.52 (µmoles H2O2 decomposed/ min/mg protein ) 9.6 ± 3.9 (U/ mg protein)

Broiler (liver)

Eraslan, et. al., 2004

Broiler (kidney)

Eraslan, et. al., 2004

Broiler (liver)

Young, et. al., 2003

GPX (U/ mg protein)

1.99 ± 0.21 39.09 ± 4.19 0.17 0.049 ± 0.016

Broiler (liver) Broiler (kidney) Broiler (liver) Broiler (liver)

Eraslan, et. al., 2004 Eraslan, et. al., 2004 Iqbal, et. al., 2002 Young, et. al., 2003

G-6-PD

59.98 ± 1.22 mU.10-9 Ec

Japanese Quail (liver)

Faix, et. al., 2003

Japanese Quail (liver) Japanese Quail (kidney) Broiler (liver) Broiler (liver)

Faix, et. al., 2003 Faix, et. al., 2003

TBARS

-1

5.3 ± 0.22 µmol.g 7.6 ± 0.21 µmol.g-1 5.3 (nmol MDA/ 100 mg) 79.23 ± 10.1 nmol MDA/g

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Young, et. al., 2003 Eid, et. al., 2003