Introduction Gout is an important metabolic disease which results due to persistent hyperuricemia and characterized by deposition of chalky urates in kidney (renal gout), joints (articular gout) or on generalized urate deposits (visceral gout)1. The disease has been widely reported in India. The principal patho-physiological change in the background of gout is considered to be either overproduction or decreased utilization of uric acid or reduced renal clearance of uric acid which leads to hyperuricemia. Uric acid constitutes major end product (70-80%) of nitrogenous catabolism in poultry. It’s less aqueous solubility makes it prone to precipitating nature. Kidney being the major excretory organ of uric acid play crucial role. |
Therefore, controlling gout is a major challenge for veterinary professionals. Poultry professionals are arbitrarily using allopurinol and febuxostat as over the counter medication to counter the problem of gout in field conditions. Allopurinol is 4-hydroxypyrazolo (3,4-d} pyrimidine. It is a purine analog, which is a structural isomer of hypoxanthine (a naturally occurring purine product in body) and an inhibitor of enzyme xanthine oxidase. It therefore, decreases uric acid formation and inhibits purine synthesis 2. Febuxostat [2- [3-cyano-4-(2-methylpropoxy) phenyl)-4-methylthiazole-5-carboxylic acid] is a non-purine selective inhibitor of xanthine oxidase. It works by non-competitively blocking the molybdenum pterincenter which is the active site on xanthine oxidase. Febuxostat inhibits both oxidized as well as reduced form of xanthine oxidase due to which febuxostat is not easily displaced from the molybdenum pterin site3. |
Though ameliorative efficacy of febuxostat and allopurinol has been assessed previously on gout induced model in broiler chicks by Patel et. al., 20174. But work on toxico-pathological study has not yet been tried in poultry. Since as an over the counter medication, most of the time they are used in higher dosage or for longer time, therefore, trial was imperative to find out whether the two anti-gout medicines which are not a part of common poultry therapeutic regimen have any toxic effect on the body system of grower broiler birds. In the present study a subacute toxicity of allopurinol and febuxostat was induced by administering 250 mg/kg b. wt. of allopurinol and 50 mg/kg b. wt. of febuxostat over a period of 7 days. The trial was imperative to find out whether these two antigout medicines which are not a part of common poultry therapeutic regimen have any toxic effect on the body system of grower broiler birds. For toxicopathological effect, that allopurinol and febuxostat could potentially cause, if any, through haematological, patho-chemical, oxidative stress enzymes, gross pathology, histopathology and ultra-structural studies. |
Top Materials and Methods Preparation of Housing Facility The trial was conducted in small animal house experimentation facility of the institute. The experimental birds of different groups were kept in different cages. Two days before the commencement of trial the entire set up was fumigated using 60 gm KMnO4 (Potassium permanganate) and 120 ml formalin per 100 cubic feet area in a closed state. The waterer and feeder were washed daily with 1% KMnO4. Management of Birds Day old, apparently healthy, eighteen Ven-cobb broiler chicks of mixed sex after acclimatization for 25 days was randomly divided into different experimental groups, namely, C group (no treatment as control), T1 group (administered allopurinol @ 250 mg/kg b. wt.) and T2 group (administered febuxostat @ 50 mg/kg b. wt.) for 7 days. The birds were fed with standard balanced ration of Uttara Food and Feeds, Venkateswara Hatcheries Pvt. Ltd. during the trial. Birds which died or sacrificed at the end of the experiment were included for the study. The details of treatment given were presented in Table 1. Collection of Blood Blood was collected from jugular vein of the experimental birds at end of the experiment period to carry out different haematological parameters. From the collected blood, two thin blood smears were prepared, dried and properly labelled with identification number of the experimental birds. 2 ml blood was transferred in BD EDTA (Ethylene Diamine Tetra acetic acid) vial containing EDTA @ 1mg/ml blood and gently mixed for haematological studies. Remaining 3 ml blood was transferred to wide mouthed BD plain vial for separation of serum for various biochemical estimations. Haematological Parameters Collected blood was used to carry out white blood cell count and total erythrocyte count using Natt-Herrick solution, as described by Natt and Herrick, 19525 as well as haemoglobin and Packed cell volume as per the method described by Schalm et. al. 19756. Haemoglobin was estimated with the help of haemoglobinometer using 450 nm filter while PCV was determined with the help of microhaematocrit centrifuge and card reader device. Erythrocyte indices were calculated as per the formulae given by Thrall, 20047. The blood smear prepared in duplicate from fresh blood samples were stained with May Grunwald-Giemsa stain following the method described by Lucas and Jamros, 1974 8 with slight modification for differential leucocyte count (DLC). Absolute count for each type of leucocyte of experimental birds was calculated as per the following formulae: Absolute count = Percentage of particular leucocyte x TLC/100 Biochemical Parameters Biochemical estimations were done from unhaemolysed serum at the end of each study using commercial test kits of M/S Tulip Diagnostics Pvt. Ltd., Ahmedabad, India. The details of procedures employed are given in Table 2. Anti-Oxidative Enzymes Serum superoxide dismutase (SOD) activity was measured spectrophotometrically using the method as described by Madesh and Balasubramanian, 19989, while serum catalase activity was estimated spectrophotometrically as per the method given by Cohen et al., 197010 with some modifications. Pathomorphological Studies Clinical Signs All the birds of different experimental groups were critically examined for the manifestation of any clinical signs in response to different treatments. Daily body weight of chicks was recorded and formulated the drug dosage accordingly. Gross and Histopathology All the organs of the necropsied experimental bird were critically examined and any gross pathological lesions observed were recorded. Tissue pieces 3-4 mm thickness were taken from kidneys, liver, heart, spleen, intestine and brain and were fixed in 10% buffered neutral formalin for histopathological studies. Sections of 4-5 µm thickness were prepared with the help of rotary microtome after they were subjected to washing, dehydration, cleaning, embedding in wax and block formation in paraffin wax. The sections were then routinely stained with Haematoxylin and Eosin and permanently mounted with DPX and cover slip for histopathological studies. Wherever necessary, special staining was done using De-Galantha stain for demonstration of urates. Ultra-Structural Pathology Representative tissue pieces of kidney collected from birds of each treated or control group were processed for transmission electron microscopy (TEM) by Technai method. Kidney tissue samples of 1.0-1.5 mm3 thickness were washed and fixed in 2.5% gluteraldehyde and 2% paraformaldehyde made in 0.1M Sodium phosphate buffer (pH 7.4) for 6 hrs. at 4°C in refrigerator. These tissue samples were then transported in 0.1M Sodium phosphate buffer for transmission electron microscopy under the aegis of electron microscopic facility (Sophisticated analytical instrument facility [SAIF], New Delhi), Department of Anatomy, All India Institute of Medical Sciences (AIIMS), New Delhi. Statistical Analysis The quantitative data of haematological observations, biochemical observations and anti-oxidative enzymes were analysed by using one-way ANOVA and Duncan’s Multiple Range Test with the help of software IBM SPSS version 20-bit 32 for showing significance between different groups as per method of Snedecor and Cochran, 196711. Ethics Approval The whole research programme was conducted after due approval of CPCSEA and institutional ethical approval. Top Results Haematological Findings The values of different haematological parameters in birds of all the three experimental groups of toxicopathological study are presented in Table 2. No significant variation in TEC was observed between different experimental groups and also, the values of TEC were found to be within normal range. A significant lowering of PCV was observed in birds of group T1 as compared to C and T2 group. The variation between C and T2 group, however was non-significant. There was no significant variation in the level of haemoglobin between different experimental groups, however, value of this parameter was found to be higher in T2 as compared to C and T1group. Highest MCV was recorded in group T2 followed by C and T1 group, though the variation between all the three experimental groups was non-significant in nature. Significantly high leucocytic count was observed in group T2 as compared to C and T1group. The TLC was also found to be higher in T1 as compared to C but the variation was non-significant. The value of absolute leucocyte count in birds of different experimental groups of toxico-pathological study is presented in Table 3. Absolute heterophil count was found to be highest in birds of group T2 followed by group C and T1. The variation between these three groups was found to be significant. Absolute lymphocyte counts of experimental birds showed significant difference in both T1 and T2 group as compared to C. The ALC of T2 was found to be lowest, which was significantly lower than T1 as well. Absolute monocyte count showed no significant variation between the three experimental groups though it was found to be highest in T2 group. A significantly high AEC was registered in the birds of group T2 as compared to group C and T1. The difference of AEC between C and T1 however, was found to be non-significant. Biochemical Findings The values of different in birds of all the three experimental groups of toxico-pathological study are presented in Table 4. There was significant reduction in uric acid level in both T1 and T2 groups as compared to C. Lowest level of uric acid was recorded in the birds of T2, though it did not differ significantly with T1. No significant variation in the level of urea was recorded between the different experimental groups. The level of serum creatinine was found to be significantly lower in T2 as compared to control group, however, it did not differ significantly from that of T1 group. The variation in serum creatinine level between groups C and T1 was non-significant. Blood glucose level did not reveal any significant variation between any of the experimental groups. However, highest glucose level was observed in group T1 and lowest in group T2. Significantly lower level of total protein was found in group T2 in comparison to control group of birds. The difference of levels of total protein in birds of group T1 with control and T2 was non-significant. The level of serum albumin was found to be significantly lower in group T2 as compared to C and T1. No significant variation was observed in serum albumin level between groups C and T1. No significant difference was recorded in the values of serum globulin between the three experimental groups, however, the value was maximum in group T2 followed by control and T1. No significant difference was recorded in the values of serum A:G ratio between the three experimental groups, however the value was more in group T2 than group T1, however, both these values were less than control group. The level of ALT was found to be significantly higher in T2 group as compared to C and T1. The level of ALT was found to be lowest in T1 though it did not differ significantly from control group. Significantly higher values of AST were recorded in group T2 as compared to control and T1 group. The difference between T1 and control group of birds was non-significant. The value of different anti oxidative enzymes in the different experimental groups of toxico-pathological study is presented in Table 5. The serum catalase and SOD activity was found to be significantly higher in both the birds of T1 and T2 group as compared to control. No significant variation of these enzymatic activities was found between group T1 and T2. Clinical Signs Birds of group T1 which received higher doses of allopurinol did not show any obvious clinical signs but one bird surprisingly showed nervous manifestation characterized by jerky movement of head and wry neck one day before completion of experiment and died suddenly. No significant clinical signs or mortality was observed in birds of T2 group which received higher doses of febuxostat. Pathomorphological Study Gross Pathology In experimentation on sub-acute allopurinol toxicity @ 250 mg/kg b.wt., the bird’s revealed nodular and uneven thickening of the wall of crop. The liver of these birds were mildly swollen and congested (Fig. 1). Mild congestion was observed in lungs also. No significant pathology or nephrosis was observed in kidneys. Brain was found to be normal. The pancreas of the affected birds gave bleached appearance. However, one bird that died during the experiment showed lesions of small ulcerative areas in the mucosa of gizzard (Fig. 2) and necrotic enteritis. Its duodenum was markedly thickened with rough irregular and uneven congested mucosal surface. Birds receiving febuxostat @ 50 mg/kg b.wt. for 7 days revealed most remarkable change in their liver which appeared yellowish brown and mildly swollen (Fig. 3). The gall bladder was distended. Cut surface of liver showed oozing of blood. Kidneys of all the experimental birds appeared grossly normal. Lungs were unaffected. The brain of few birds was mildly congested. No remarkable pathology was observed in spleen, bursa or thymus. The intestinal wall was almost normal except the wall of oesophagus which showed mild thickening. In few birds, the wall of crop was markedly thickened and oedematous. In one bird large ulcer crater was observed in the crop mucosa (Fig. 4). In the birds of control group, no significant gross pathology was observed in any of the organs. Histopathology Normal histological features were observed in most of the kidney of allopurinol subacute toxicity group (T1). At few places there were hypercellularity of mesangial cells in glomerulus and dilatation of capillaries (Fig. 5). At some places interstitial nephritis was observed characterized by infiltration of inflammatory cells particularly mononuclear cells in the interstitial space of medullary region. In group T2, major lesions were observed in glomerular tuft instead of tubules. Glomerular tufts were enlarged and mildly hypercellular. The capillaries were dilated. Often there was obliteration of the bowman’s space and bridging between parietal and visceral layer of Bowman’s capsule (Fig. 6). Proteinaceous exudates were found in the tubular lumen of cortical areas. No remarkable histological change was seen in liver due to allopurinol subacute toxicity except mild portal fibrosis. The portal blood vessels were congested. The hepatocytes appeared normal. The birds of group T2 showed significant dilatation of sinusoidal space and atrophy of hepatocytes with presence of pyknotic nuclei in few hepatocytes. The most remarkable change was significant portal fibrosis and was of increased severity as compared to birds of group T1 (Fig. 7). The fibrous connective tissue in the portal area of the birds of group T2 showed tendency to infiltrate into the lobules of the liver. Congestion and few areas of focal necrosis were also observed. No major histopathological alteration was observed in the brain of the birds of T1 group except mild congestion of blood vessels. Meningitis with infiltration of mononuclear cells was observed in birds of T2 group (Fig. 8). Mild oedematous and congestive changes were also observed in sub-meningeal region. Significant pathology was observed in the villi of intestinal mucosa in the birds of T1 group. There was marked hyperplasia of goblet cells. Thickening and club shaped blunting of the tip of villi was also observed (Fig. 9). Marked fusion of the villi and thick coating of mucous over the tip was also recorded. Apical part of villi showed degeneration and necrosis. Intracellular oedema was seen in the cells at tip of the villi. At places, there was fusion of intestinal villi into a clumped mass. The degenerated enterocytes were pooled into a lobular pattern. The villi of birds in T2 group appeared normal with mild hyperplasia of goblet cells and mild degenerative changes in the epithelial cells at tip of villi (Fig. 10). No abnormal histological alteration of significance was observed in birds of any of the experimental groups. Lungs in all the experimental groups appeared histologically normal. Ultrastructural Pathology In group T1 no significant ultrastructural changes were seen in the renal tubules of birds administered with higher dose of allopurinol to produce sub-acute toxicity (Fig. 11, 12). In group T2, the kidneys of birds in this group showed huge number of small mitochondria in some places almost completely packing the cytoplasm (Fig. 13). Nuclear membrane was intact (Fig. 14). Electron dense bodies were absent in the tubular cells. Top Discussion Toxicity study is also carried out to determine an adverse effect level. Thus, the present study on toxic effects of allopurinol and febuxostat is aimed at providing preliminary toxico-pathological effect of these agents which could be integrated with detailed pharmokinetic and toxicokinetic effects of these agents in further research work to develop an ideal therapeutic dose for poultry grower birds of allopurinol and febuxostat as an antigout agent. |
A comparative haematological analysis of allopurinol and febuxostat toxicity clearly showed that except for development of significant yet mild heterophilic leucocytosis in the febuxostat intoxicated birds (T2) these agents had absolutely no effect on haematopoiesis. |
It was interesting to note that though ten times of the therapeutic dose of both allopurinol and febuxostat has been used to induce subacute toxicity no significant alteration in the level of serum uric acid, urea, creatinine, total protein, albumin, globulin and A:G ratio was observed in both T1 and T2 group as compared to control birds. However, the liver function test parameter of ALT and AST showed higher enzymic level in T2 group as compared to T1 and control. Both of T1 group however showed normal levels for all biochemical parameters. This clearly indicates that allopurinol is a much safer and less toxic medicine for poultry. Febuxostat toxicity on the other hand, showed significant rise in the enzymic levels of ALT and AST i.e. the parameters for liver function. This brings to the fore hepatotoxic potential of febuxostat when it is used in over dose. |
Another interesting finding was the observations of serum uric acid which showed significant reduction in its level in both T1 and T2 birds. This lowering of serum uric acid level in treatment groups T1 and T2, however may have a negative impact on the health of the birds. This assumption is based on the fact that uric acid has proven antioxidant properties and lowering in the level of uric acid beyond permissible level might have made experimental birds of T1 and T2 susceptible to injury by oxygen derived free radicals, the generation of which is a common phenomenon in inflammatory reaction. |
The level of catalase and SOD enzymes were significantly increased in both the toxicity groups indicating oxidative stress due to their deleterious effect on various body organs. |
Detailed pathological examination of the birds of both the toxicity group at the end of seven-day intoxication reveal significant changes. Allopurinol toxicity lead to ingluvitis characterized by thickening of the wall of crop and mild degree of ulcerative ventriculitis characterized by formation of small ulcerative areas on the mucosa of gizzard. Duodenal wall also showed marked thickening with rough irregular mucosal surface. Thickening and oedematous changes in wall of oesophagus, crop and development of ulcerative ingluvitis in one bird in T2 group suggests that both allopurinol and febuxostat had corrosive effect on the upper digestive tract of broiler grower birds. |
Febuxostat toxicity showed significant pathology in liver, however kidney mostly remain unaffected and did not reveal any significant degenerative changes. The toxic effect of febuxostat or its metabolites must have influenced water and sodium ion influx which expands the membranous compartments of mitochondria, lysososmes and endoplasmic reticulum to produce hydropic degeneration of hepatocytes. However, these retrogressive changes were mild in nature as we evidenced that only mild increase in the level of liver specific enzyme ALT and AST was recorded in the present study. The liver specific enzyme level in allopurinol intoxicated birds was well within the normal range. Hence, we can conclude that allopurinol is nontoxic to liver and febuxostat is mildly toxic to the liver of broiler grower birds. |
The other major histopathological finding of bile duct hyperplasia and portal fibrosis was observed only in febuxostat intoxication only. Since there was no cholestasis in the present study, hepatocytes showed hydropic degeneration and no inflammatory cell infiltration was observed in portal triad, the bile duct proliferation may have developed in response to portal fibrosis which was clearly observed in febuxostat intoxicated birds. Bile duct proliferation may occur quite independently of changes in the parenchyma, particularly when an irritant stimulus is considered typical response to toxins such as alphanaphthylisothiocyanate12. |
In the present studies on febuxostat toxicity major fibrosis was observed in the portal area where collagen type I and III predominates. This is suggestive of the fact that primarily there was development of portal/biliary fibrosis, most probably due to direct toxic effect of febuxostat reaching the site by entero-hepatic circulation or through systemic circulation to induce excessive EMC deposition in the form of fibrous connective tissue. Other possible route of portal fibrosis may be following metabolism of febuxostat in the hepatocytes and access of toxic/irritant metabolite of febuxostat getting excreted through bile and reaching portal area to initiate biliary fibrosis. The second pathway appears less likely as no cholestatic changes were observed in the present study. The proliferating nature of fibrous connective tissue into the hepatic lobules along with increased connective tissue in the sinusoidal space and proliferation and rounding of kupffer cells is strongly suggestive of the fact that there is secondary involvement of sinusoidal space from interstitial collagen production. It also gives strong indication of the fact that if febuxostat toxicity is persisted for longer duration then liver may show increased susceptibility for frank cirrhosis and hepatic failure. |
These findings provide ample opportunity to explain in greater details the true nature of hepatotoxic potential of febuxostat. Whether continuation of intoxication can lead to progressive fibrosis and cirrhosis or these fibrotic changes are reversible in nature, if the toxicity with febuxostat is withdrawn. It is important to mention here that hepatic fibrosis is considered potentially reversible, if the underlying insult is effectively treated or stopped, apoptosis of hepatic stellate cells and reduced expression of metaloproteinase inhibition slow down the production of ECM and allow degradation of immature collagen by matrix metaloproteinases to proceed. The outcomes are usually considered favourable for progressive fibrosis when insult continues to act and collagen maturation and cross linking potentially reduce its susceptibility to enzymic degradation. |
Thus, we can conclude from toxicity study that allopurinol has greater safety margin in broiler grower birds whereas febuxostat toxicity carry inherent capability of causing hepatic necrosis, progressive portal fibrosis, cirrhosis and end stage hepatic failure if the broiler grower birds are subjected to long term febuxostat toxicity. Similar observations in human study were made by some previous workers13,14. |
How to cite this article : Kumar, S., Gupta, M.K. and Kumar, S. 2022. Comparative toxicopathological studies on sub-acute toxicity of allopurinol and febuxostat against induced hyperuricemia in broiler model. Indian J. Vet. Pathol., 46(4) : 303-310. Top Figures | Fig. 1.: Photograph showing swollen liver in bird of T1 group
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| | Fig. 2.: Photograph showing small ulcers in gizzard of a bird of T1 group
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| | Fig. 3.: Photograph showing yellowish brown liver in a bird of T2 group
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| | Fig. 4.: Photograph showing crater of large ulcer in the crop of bird of T2 group.
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| | Fig. 5.: Photomicrograph of kidney showing hyper-cellularity of mesangial cells and dilatation of glomerular capillaries in birds of T1 group. (H&E ×1000)
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| | Fig. 6.: Photomicrograph of kidney showing bridging between parietal and visceral layer of Bowman’s capsule in birds of T2 group. (H&E ×1000)
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| | Fig. 7.: Photomicrograph of liver showing portal fibrosis in birds of T2 group. (H&E ×100)
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| | Fig. 8.: Photomicrograph of brain showing mild infiltration of inflammatory cells in the meninges of birds of T2 group. (H&E ×100)
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| | Fig. 9.: Photomicrograph of intestine showing thickening and club shaped blunting of the tip of villi in a bird of T1 group. (H&E ×400)
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| | Fig. 10.: Photomicrograph of intestine showing degenerative changes in the epithelial cells at tip of villi in birds of T2 group. (H&E ×400)
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| | Fig. 11.: Photomicrograph showing TEM view of kidney showing almost normal appearance in a bird of T1 group. (×2550)
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| | Fig. 12.: Photomicrograph showing TEM view of kidney showing almost normal appearance of tubular epithelial cell in a bird of T1 group. (×2550)
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| | Fig. 13.: Photomicrograph showing TEM view of huge number of mitochondrial presence in the renal epithelial cells of a bird of T2 group. (×2550)
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| | Fig. 14.: Photomicrograph showing TEM view of intact nuclear membrane in the renal tubular epithelial cells of a bird of T2 group. (×2550)
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Tables | Table 1.: Experimental groups and treatment schedule for toxico-pathological study.
| | Groups | No. of birds and age | Treatment | Duration | Route | | C | 6 (25 day old) | No Treatment | 7 days | per os | | T1 | 6 (25 day old) | Allopurinol @ 250 mg/kg b.wt. | 7 days | per os | | T2 | 6 (25 day old) | Febuxostat @ 50 mg/kg b.wt. | 7 days | per os |
| | | Table 2.: Haematological parameters (Mean ± S.E.) in the different experimental groups of toxico-pathological study.
| | Parameters/Groups | C | T1 | T2 | | TEC (m/mm3) | 3.18±0.05a | 3.15±0.09a | 3.22±0.14a | | PCV (%) | 34.50±0.42b | 32.66±0.61a | 34.83±0.97b | | Hb (g%) | 10.90±0.21a | 10.88±0.25a | 11.42±0.16a | | MCV (fl) | 108.54±2.31a | 104.14±3.71a | 109.45±5.47a | | MCH (pg) | 34.28±0.78a | 34.69±1.33a | 35.86±1.78a | | MCHC (gm/dl) | 31.61±0.68a | 33.30±0.20b | 32.77±0.19ab | | TLC (/mm3) | 10383.33±218.20a | 11016.67±175.91a | 12716.67±298.24b | | Differential Leucocyte Count | | Heterophil | 37.83±2.06a | 47.16±1.72b | 56.50±0.67c | | Lymphocyte | 54.17±1.74c | 45.00±2.08b | 34.50±0.43a | | Monocyte | 3.00±0.37a | 2.84±0.31a | 3.17±0.48a | | Eosinophil | 5.00±0.45a | 5.00±0.52a | 5.83±0.48a |
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| Means bearing different superscript in a column differ significantly. | | | Table 3.: Absolute leucocyte count (Mean ± S.E.) in the different experimental groups of toxico-pathological study.
| | Parameters/Groups | C | T1 | T2 | | AHC | 3928.01±134.31a | 5195.46±217.87b | 7184.91±201.76c | | ALC | 5624.64±134.31c | 4957.50±209.46b | 4387.25±98.21a | | AMC | 311.49±35.56a | 312.87±35.34a | 403.11±59.70a | | AEC | 519.16±38.61a | 550.83±62.24a | 741.38±67.93b |
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| Means bearing different superscript in a column differ significantly. | | | Table 4.: Biochemical parameters (Mean ± S.E.) in the different experimental groups of toxico-pathological study.
| | Parameters/Groups | C | T1 | T2 | | Uric acid (mg/dl) | 5.48±0.12b | 4.00±0.06a | 3.82±0.15a | | Urea (mg/dl) | 13.63±1.08a | 14.09±0.49a | 12.66±0.44a | | Creatinine (mg/dl) | 0.35±0.02b | 0.32±0.02ab | 0.27±0.02a | | Glucose (mg/dl) | 205.35±2.34a | 210.45±3.16a | 201.68±2.74a | | Total protein (mg/dl) | 4.40±0.09a | 4.27±0.10a | 4.22±0.08a | | Albumin (mg/dl) | 1.68±0.05b | 1.68±0.04b | 1.53±0.02a | | Globulin (mg/dl) | 2.71±0.12a | 2.58±0.35a | 2.68±0.09a | | A:G | 0.62±0.04a | 0.66±0.49a | 0.57±0.02a | | ALT (IU) | 24.17±2.71a | 20.00±2.58a | 39.17±3.27b | | AST (IU) | 28.33±3.33a | 26.67±3.07a | 56.67±4.94b |
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| Means bearing different superscript in a column differ significantly. | | | Table 5.: Anti-oxidative enzymes count (Mean ± S.E.) in the different experimental groups of toxico-pathological study.
| | Parameters/Groups | C | T1 | T2 | | Catalase (IU) | 3.85±0.04a | 6.13±0.11b | 6.28±0.07b | | SOD (IU) | 150.67±0.99a | 180.83±1.42b | 182.67±0.95b |
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| Means bearing different superscript in a column differ significantly. | |
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