Clinico-pathological studies and therapeutic effects of febuxostat on visceral gout in vanaraja chicken

Introduction

India's poultry industry is one of the fastest-growing sectors within agriculture, significantly contributing to the national economy. Among various poultry breeds, Vanaraja is a dualpurpose bird, suitable for both egg and meat production, particularly under rural and backyard farming conditions. However, with advancements in genetic selection for higher productivity, poultry birds are increasingly prone to metabolic disorders, including visceral gout-a major cause of economic losses in the industry. Visceral gout is a metabolic disorder characterized by urate deposits on visceral organs due to elevated blood uric acid levels. It is often linked to nutritional imbalances, management issues, nephrotoxic drugs, and infectious agents. Studies suggest that veterinary use of nonsteroidal anti-inflammatory drugs (NSAIDs), particularly aceclofenac, may contribute to visceral gout, similar to diclofenac's known toxicity in vultures.

Febuxostat, a selective xanthine oxidase inhibitor, has shown promise in lowering serum uric acid levels by inhibiting its synthesis, offering a potential therapeutic approach for managing visceral gout in poultry. Additionally, natural remedies like bottle gourd (Lagenaria siceraria) possess medicinal properties that may support kidney function and metabolic health. Despite the economic significance of visceral gout in poultry, limited research has been conducted on its prevalence and management in Vanaraja chickens, especially in Bihar. This study aims to investigate the role of aceclofenac in visceral gout, evaluate the therapeutic potential of Febuxostat and analyze the associated haemato-biochemical.

Materials and Methods

Experimental Design

The study on "Clinico-Pathological Studies and Therapeutic Effect of Febuxostat on Visceral Gout in Vanaraja Chickens" was conducted at ILFC, BVC, Patna, and the Department of

Packed Cell Volume (PCV)

Packed cell volume (PCV) is a critical hematological parameter used to assess dehydration (increased PCV), anemia (decreased PCV), polycythemia (increased PCV), and overall nutritional status. The mean PCV values of Vanaraja chicks across different experimental groups. The increase in TLC in aceclofenac-induced groups may be due to metabolic acidosis leading to uremia, which could stimulate bone marrow activity, resulting in leukocytosis.

The protective group (Group III) did not show significant changes, suggesting febuxostat's role in stabilizing immune responses.

Hematological-Parameters

Hemoglobin levels remained unchanged across groups at day 7. By day 14, Groups II and IV showed significantly elevated Hb levels compared to the control (p<0.05), indicating dehydration and impaired kidney function. However, Groups I and III maintained stable Hb values throughout the experiment, suggesting that febuxostat prevented significant hematological alterations.

The study confirms that aceclofenac toxicity induces visceral gout, weight loss, and hematological alterations in Vanaraja chickens. Both bottle gourd juice and febuxostat exhibited protective effects by reducing clinical signs, improving weight gain, and stabilizing hemoglobin levels. These findings highlight their potential role in mitigating aceclofenac-induced toxicity in poultry.

Differential Leukocyte Count (DLC) Lymphocytes

The Mean ± S.E values of lymphocytes in Vanaraja chicks of Group I, II, III, and IV were recorded at different time intervals. No significant alterations were observed in all groups at the end of 7 days. However, at the end of 14, 21, and 28 days, the aceclofenac-induced Groups II and IV exhibited comparatively higher lymphocyte counts than Groups I and III. The lymphocyte count in Group III (aceclofenac + febuxostat) did not show any significant difference when compared to the control Group I. By the end of 42 days, no significant differences were observed among the groups, indicating a reversal of lymphocyte elevation. The increased lymphocyte count on days 14, 21, and 28 in aceclofenac-induced groups aligns with findings by Christopher (1978) [2] , who also reported elevated lymphocyte counts in gout-affected birds. The observed increase might be due to metabolic acidosis leading to uremia, which could have stimulated the bone marrow to produce more lymphocytes as a compensatory mechanism.

Heterophils

The Mean ± S.E values of heterophils showed no significant alterations in any of the groups throughout the experimental period (7, 14, 21, 28, and 42 days). Normal heterophil counts indicate the absence of any significant infection in Vanaraja chickens. This observation is in agreement with studies conducted by Koutsos et al. (2020) [1] and Christopher (1978) [2] , who also reported no significant changes in heterophil counts.

Eosinophils

The eosinophil counts remained unchanged in all experimental groups at the end of 7, 14, 21, 28, and 42 days. This suggests that the birds did not have any parasitic infections, further supporting findings from Christopher (1978) [2] and Koutsos et al. (2020) [1] , who observed no significant variations in eosinophil counts.

Monocytes

The Mean ± S.E values of monocytes in all experimental groups remained stable across all time points. The normal monocyte count indicates the absence of significant infections or inflammatory responses in the experimental birds. These findings align with the reports of Christopher (1978) [2] , Koutsos et al. (2020) [1] , and Singh et al. (2013) [3] , who also noted no significant changes in monocyte counts.

Basophils

Similar to other leukocyte parameters, no significant alterations were observed in basophil counts across all experimental groups at different time intervals. The normal basophil counts indicate the absence of allergic reactions or infections in Vanaraja chickens, as also reported by Christopher (1978) [2] , Koutsos et al. (2020) [1] , and Singh et al. (2013) [3] .

Serum Biochemical Parameters

Blood samples were collected from all experimental groups at the end of 7, 14, 21, 28, and 42 days. Serum was separated and analyzed for biochemical parameters.

Serum Uric Acid

The Mean ± S.E values of serum uric acid in different groups indicated significant changes in aceclofenac-induced groups. At the end of 7 days, no significant differences were observed among the groups. However, at the end of 14, 21, and 28 days, Groups II and IV (aceclofenac-induced) exhibited significantly higher serum uric acid levels compared to Groups I and III. Notably, Group III (aceclofenac + febuxostat) did not show any significant deviation from the control Group I. The elevation of serum uric acid in Groups II and IV is indicative of impaired renal function due to aceclofenac administration, leading to hyperuricemia and visceral gout. These findings are in agreement with previous studies by Gajera (2006) [4] , Shinde (2007) [5] , Patel et al. (2007) [6] , Sharma et al. (2012) [8] , Ghodasara et al. (2014) [9] , and Ramjan et al. (2015) [10] , who reported increased uric acid levels in diclofenac-treated birds. Similar observations were made by Chandra et al. (1985) [11] , Mert (1991) [12] , Uma et al. (1997) [13] , Oaks et al. (2004) [14] , Irtaza et al. (2008) [15] , Hedaoo et al. (2008) [16] , and Sharma and Vegad (2010) [8] . The increased uric acid levels observed in gout-affected birds could be attributed to aceclofenac-induced renal damage, leading to inefficient uric acid excretion. The accumulation of uric acid resulted in supersaturation of blood and subsequent deposition in visceral organs, causing inflammatory changes and gout. By the end of 42 days, Groups II and IV, which were treated with bottle gourd juice and febuxostat, respectively, exhibited significant reductions in serum uric acid levels.

These findings indicate that both bottle gourd juice and febuxostat played a protective role in mitigating hyperuricemia. The effectiveness of febuxostat in reducing serum uric acid levels has been well-documented by Horiuchi et al. (1999) [17] , Khosravan et al. (2006) [18] , and Komoriya et al. (2004) [19] , who reported a dose-dependent reduction in serum uric acid following febuxostat administration.

Additionally, bottle gourd juice demonstrated a significant reduction in serum uric acid levels, aligning with findings from Kausar and Waris (2016) [20] , who reported that bottle gourd effectively facilitated uric acid excretion in hyperuricemic rats. In the present study, a 20 ml dose of bottle gourd juice was found to effectively reduce serum uric acid levels in Vanaraja birds.

Serum Creatinine

Creatinine analysis is an important biochemical diagnostic tool to assess kidney function. It is primarily eliminated from the blood via glomerular filtration and proximal tubular secretion in the kidneys. The elevated creatinine levels in Groups II and IV at Days 14, 21, and 28 indicate that aceclofenac induces kidney dysfunction. These findings align with previous studies by Feizi et al. (2012) [21] , Irtaza et al. (2008) [15] , Shinde (2007) [5] , Hedaoo et al. (2008) [16] , Jain et al. (2009) [22] and Undhad et al. (2012) [23] . The administration of bottle gourd juice in Group II and febuxostat in Group IV demonstrated a protective effect against aceclofenac-induced nephrotoxicity. A significant reduction in serum creatinine levels was observed after 28 and 42 days. This suggests that both treatments effectively mitigate kidney damage, with bottle gourd juice playing a crucial role in renal protection by eliminating excess uric acid, as supported by Kausar and Waris (2016) [20] .

Serum Total Protein (TSP)

The Mean ± S.E values of serum total protein in Vanaraja chicks of Groups I, II, III, and IV at different time intervals revealed no significant alterations at Days 7, 14, 21, 28, and 42. The normal TSP values indicate the absence of systemic infections in the birds. This finding is consistent with the studies of Singh et al. (2013) [3] and Chandra et al. (1985) [11] , who reported no significant changes in serum total protein under similar conditions.

Serum Albumin

The The increased serum albumin levels in Groups II and IV from Days 14 to 28 suggest that aceclofenac toxicity contributes to elevated albumin levels, possibly due to dehydration. These findings align with the reports of Chandra et al. (1985) [11] and Singh et al. (2013) [3] . The administration of bottle gourd juice in Group II and febuxostat in Group IV resulted in a significant reduction in albumin levels by Days 28 and 42, demonstrating their protective effects against aceclofenac-induced visceral gout.

The results indicate that both treatments helped restore albumin concentrations to near-normal levels, mitigating the impact of aceclofenac-induced nephrotoxicity.

Serum Globulin

The [11] and Singh et al. (2013) [3] , who attributed decreased globulin levels to emaciation and dehydration caused by drug-induced toxicity. Interestingly, by the 42 nd day, serum globulin levels in Group II (treated with bottle gourd juice) and Group IV (treated with febuxostat) showed a marked recovery, reaching levels comparable to the control group. This suggests a potential protective role of bottle gourd juice and febuxostat in mitigating the adverse effects of aceclofenacinduced visceral gout. The findings reinforce the hepatoprotective and nephroprotective properties of these treatments in restoring normal globulin concentrations.

Serum Glutamic Pyruvic Transaminase (SGPT)

Serum SGPT, also known as alanine transaminase (ALT), serves as an essential biomarker for liver function assessment. The mean ± S.E. values of serum SGPT at various time points are presented in Table 2. At the end of 7 days, no significant differences were observed among the groups. However, by the 14th day, aceclofenac-induced Groups II and IV exhibited significantly elevated SGPT levels compared to the control and preventive group. This trend persisted through 21 and 28 days, indicating aceclofenac's detrimental effect on liver function.

The increased SGPT levels in aceclofenac-induced groups suggest hepatocellular damage, likely due to inflammation or oxidative stress. These findings align with previous studies by Chandra et al. (1985) [11] , Garcia (1998) [24] , and Jain et al. (2009) [22] , who reported elevated ALT levels in cases of drug-induced hepatotoxicity. Darbar et al. (2010) [25] and Kausar and Waris (2016) [20] also observed increased ALT levels in experimental models of uric acid-induced gout.

By the end of 42 days, Groups II and IV, which were treated with bottle gourd juice and febuxostat, respectively, demonstrated a significant reduction in SGPT levels, reaching values comparable to the control group. This suggests that both interventions effectively counteracted the hepatotoxic effects of aceclofenac. Patel (2017) [5] similarly reported a significant decrease in serum ALT levels in animals treated with febuxostat, reinforcing its hepatoprotective potential. Additionally, bottle gourd juice demonstrated a notable effect in restoring liver function by significantly reducing SGPT levels after 28 and 42 days of treatment.

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