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Evaluation of Antifasciola Efficacy of Artemisia Maritima in Relation to Closantal and Triclabendazole in Bovines Affected With Fasciolosis

Imtiyaz Ahmad Reshi Rajiv Singh A. H. Bhat S. R. Upadhyay I. A. Shah
Vol 7(5), 134-139
DOI- http://dx.doi.org/10.5455/ijlr.20170330052752

Parasitism in livestock is a cause of concern for animal health and production. Among helminth infections, fasciolosis is of paramount importance due to its wide host range. Resistance of flukes against commonly available flukicides has been reported from various parts of the world. To overcome this limitation and dependency over the limited anthelmintics, herbal preparation, Artemisia maritima was used to treat this infection. The in- vivo trial was conducted on 18 bovines (divided in 3 groups, G1, G2 and G3) naturally infected with Fasciola spp. G1 animals were treated with Artemisia maritima powder @750 mg/kg BW orally, G2 and G3 animals received a single oral dose of closantel @7.5 mg/kg BW and triclabendazole @12 mg/kg BW orally, respectively. Each group was monitored for a period of 21 days post treatment. Six infected animals were kept as infected untreated (positive) control (G-4) and same number of apparently healthy animals free from endoparasitic infestation was kept as uninfected and untreated (negative) control. Pre and post-treatment values of biochemical and mineral parameters in Fasciola infected bovine was estimated. Triclabendazole and closantel showed significant (p<0.05) increase in TPP, albumin, A: G ratio, AST, Ca and P levels 21 days after treatment whereas, Artemisia maritima treated group showed non-significant improvement.


Keywords : Fasciolosis Artremisia Closental Triclabendazole

Introduction

Parasitism in livestock is a cause of concern for animal health and production. Huge economic losses occur due to reduced fertility, lowered milk yield and decreased meat production (Torgerson and Claxton, 1999; Schweizer et al., 2005). Among helminth infections, fasciolosis is of paramount importance due to its wide host range (Rondelaud et al., 2001). In bovines, fasciolosis occurs in both acute and chronic form, chronic being the primary form. Acute fasciolosis, caused by immature flukes leads to anaemia, hypoproteinemia and high mortality due to impaired hepatic functions (Soulsby, 1987). Chemotherapy is the only tool available to control this condition, but their effectiveness needs to be evaluated under field conditions. Resistance of flukes against commonly available flukicides has been reported from various parts of the world (Brennan et al., 2007). Therefore, to overcome this limitation and dependency over the limited anthelmintics, some herbal preparations seem to provide an alternative for effective and cheaper remedy. Keeping this mind, Artemisia maritima was used to treat this disease. Artemisia maritima grows in high altitudes of Kishtwar area of Jammu region and is a promising anthelmintic in human use. Accordingly, the present study was undertaken to evaluate the therapeutic efficacy of closantel and triclabendazole in comparison to Artemisia maritima in bovines naturally infected with Fasciolia.

Materials and Methods

The comparative anti-Fasciola efficacy of closantel, triclabendazole and Artemisia maritima L. Ex Hookf. was tested in bovines naturally suffering with Fasciola spp. The in- vivo trial was conducted on 18 bovines naturally infected with Fasciola spp. The infected animals were randomly divided into three groups (G 1, G 2 and G 3). The six infected animals were kept as infected untreated (positive) control (G-4) whereas, same number of apparently healthy animals free from endoparasitic infestation were kept as uninfected and untreated (negative) control. G 1 animals were treated with Artemisia maritima powder at rate of 750 mg/kg BW orally; G 2 and G 3 animals received a single oral dose of closantel at rate of 7.5 mg/kg BW and triclabendazole at rate of 12 mg/kg BW orally, respectively. Each group was monitored for a period of 21 days post treatment (DPTs). The therapeutic efficacy of plant extract and drugs was assessed on the basis of reduction in eggs per gram of faeces (EPG) count and improvement in biochemical parameters.

The faecal samples were collected in morning hours on weekly basis directly from the rectum of animals over the period of 21 days (the period of experiment). The samples were immediately analysed by direct smear and Stoll’s technique to estimate the number of EPG. Approximately 30 ml of blood sample was collected from each selected animal in mineral free glass vials (dipped overnight in 2 N HCl) on 0, 14th and 21st day of trail. Plasma was separated and total plasma protein (TPP) and albumin levels were estimated using Biuret Method (ERBA® kits); aspartate aminotransferase (AST) and Alanine aminotransferase (ALT) were estimated by IFCC method (ERBA® kits). The calcium and inorganic phosphorous (Pi) were analysed from plasma samples employing OCPC method (ERBA® kits). Trace minerals i.e., iron and copper were analysed using Zeeman Atomic Absorption Spectrophotometer (Z-2300, HITACHI). Statistical analysis was done using Duncan multirange test as per method of Snedecor and Cochran, 1994.

Results and Discussion

Analysis of plasma samples from naturally Fasciola infected animals revealed significant (p< 0.05) increase in level of hepatic enzymes i.e., aspartate aminotransferase (54.11± 0.43 IU/L) and alanine aminotransferase (21.98± 0.58 IU/L) compared with healthy control group having average values 29.17± 0.19 IU/L and 17.68± 0.67 IU/L, respectively (Table 1).

Table1: Plasma biochemical and mineral profile of healthy and Fasciola infected bovines (Mean ± S.E.)

Parameters Healthy Control

(n=6)

Infected Group

(n=18)

T. Protein (g/dl) 5.88± 0.06 a 4.43± 0.07b
Albumin (g/dl) 3.41± 0.06 a 1.95± 0.04b
A: G 1.29± 0.04 a 0.83± 0.03b
AST (IU/L) 29.17± 0.19 a 54.11± 0.43b
ALT (IU/L) 17.68± 0.67 a 21.98± 0.58b
Ca (mg/dl) 8.30±0.03a 6.63±0.09b
Pi (mg/dl) 4.96±0.08a 3.45±0.04b
Fe (µmol/l) 31.84±0.48a 21.73±1.01b
Cu (µmol/l) 12.35±0.10a 9.80±1.70a

Values with different superscript in rows differ significantly (p< 0.05)

Disturbance in hepatic enzyme level can be attributed to the liver damage caused by immature and mature liver flukes which is in agreement with the earlier findings of Edith et al. (2012); Sheikh et al. (2006) and Chakraborty (1999). Fasciola infected animals showed significant (p<0.05) decrease in TPP (4.43 g/dl), albumin (1.95 g/dl) and A: G values (0.83± 0.03) compared to healthy control animals (average values 5.88 g/dl, 3.41 g/dl and 1.29± 0.04, respectively). The decrease in these biochemical constituents might be due to the reduced synthesis in damaged liver and increased plasma leakage into the gut (Radostits et al., 2007). Similar findings have been reported by Sheikh et al. (2006) and Al-Saffar (2008) in cattle and sheep, respectively infected with fasciolosis. Ca and Pi level of Fasciola infected animals showed significant decline from 8.30± 0.03 mg/dl and 4.96± 0.08 mg/dl among healthy control group to 6.63± 0.09 mg/dl and 3.45± 0.04 mg/dl among Fasciola infected animals, respectively. Decreased absorption from gut, diarrhoea and decreased food intake in the infected animals could be responsible for lowered levels. Finding corroborates with the observation of Sheikh et al. (2006) and Abouzeid et al. (2010). Significant (p<0.05) decrease in plasma Fe level was recorded which corroborates with the findings of El-Khadrawy et al., 2008; Lotfollahzadeh et al., 2008 and Abouzeid et al., 2010 in humans, buffalo-cows and cattle and sheep, respectively affected with fasciolosis. Decrease in iron values may be attributed to blood sucking activity of flukes. A non-significant (p>0.05) decline in plasma copper level of Fasciola infected animals was observed compared to healthy control group. Contrary to present findings El-Khadrawy et al. (2008) and Abouzeid et al. (2010) reported a significant decrease in copper level of Fasciola infected buffaloes, cows and sheep and attributed it to liver damage caused by flukes as it is the major storage organ. Following treatment with triclabendazole and closantel significant (p<0.05) increase in TPP, albumin, A:G ratio, AST, Ca and Pi levels was observed 21 days after treatment whereas, Artemisia maritima treated group showed non-significant improvement. Marked improvement in biochemical parameters have also been recorded by Sheikh et al. (2006), El-khadrawy et al. (2008) and Pal and Dasgupta (2006).

Table 2: Pre and post-treatment values of biochemical parameters in Fasciola infected bovine

(Mean ±SE)

S. No. Parameters Group Time Interval Following Treatment
Day 0 Day 14 Day 21
1. TPP (g/dl) 1 4.37± 0.05aA 4.24± 0.05aA 4.08± 0.03bA
2 4.53± 0.21aA 4.63± 0.03aB 5.65± 0.04bB
3 4.38± 0.07aA 4.55± 0.04aB 5.72± 0.06bB
4 4.53± 0.13aA 4.11± 0.01aA 3.68± 0.02bA
5 5.88± 0.06aB 5.98± 0.23aC 5.94± 0.23aC
2. Albumin (g/dl) 1 1.92± 0.07aA 1.81± 0.06abA 1.70± 0.06bA
2 1.98± 0.07aA 2.19± 0.04aB 2.89± 0.09bB
3 1.94± 0.07aA 2.11± 0.04aB 2.92± 0.06bB
4 1.95± 0.06aA 1.76± 0.05aA 1.42± 0.03bA
5 3.41± 0.06aB 3.43± 0.01aC 3.48± 0.02aC
3. A:G ratio 1 0.76± 0.04aA 0.76± 0.07aA 0.75± 0.14aA
2 0.82± 0.07aA 0.89± 0.06aA 0.94± 0.05bB
3 0.90± 0.06aA 0.92± 0.06aB 1.03± 0.08bBC
4 0.75± 0.05aA 0.74± 0.06aA 0.61± 0.04bA
5 1.28± 0.04aB 1.34± 0.06aC 1.41± 0.01aC
4. AST (IU/L) 1 53.33± 0.67aA 53.17±0.40aBC 53.67± 0.42aC
2 55.17± 0.60aA 54.67± 0.49aC 49.19± 0.60bB
3 53.83± 0.83aA 52.17± 0.48aB 48.17± 0.94bB
4 54.20± 0.71aA 60.81±0.8aBC 60.62±0.93bD
5 29.17± 0.19aB 29.43±0.14aA 28.67±0.12aA
5. ALT (IU/L) 1 20.72± 0.79aA 20.72± 0.93aB 20.65± 0.75aB
2 22.15± 0.98aA 20.88± 0.88aB 19.65± 0.90aB
3 23.08± 1.15aA 21.47± 1.01aB 19.85± 0.97aB
4 21.94± 0.13aA 22.27± 0.16aB 23.12± 0.21aC
5 17.68± 0.67aB 17.63± 0.57aA 17.54± 0.43aA

Values with different superscript (small letters in rows and capital letters in columns) differ significantly (p< 0.05)

Table 3: Pre and post-treatment values of mineral parameters in Fasciola infected bovine (Mean ±SE)

S. No. Parameters Group Time Interval Following Treatment
Day 0 Day 14 Day 21
1. Ca (mg/dl) 1 6.57±0.16aA 6.47±0.15aA 6.25±0.15aA
2 6.62±0.13aA 6.95±0.16aB 7.73±0.12bB
3 6.70±0.17aA 7.09±0.16aB 8.02±0.10bBC
4 6.61± 0.17aA 6.54± 0.12aA 6.47± 0.11aA
5 8.30± 0.03aB 8.29± 0.01aC 8.31± 0.03aC
2. Pi (mg/dl) 1 3.43±0.10aA 3.39±0.08aA 3.34±0.12aA
2 3.42±0.04aA 3.54±0.02aB 4.91±0.04bB
3 3.50±0.01aA 3.62±0.08aB 4.93±0.02bB
4 3.47± 0.05aA 3.43± 0.04aA 3.36± 0.11aA
5 4.96± 0.08aB 4.93± 0.03aC 4.94± 0.01aB
3. Fe (µmol/l) 1 21.56± 2.2aA 20.31± 1.14aC 19.05± 1.08aC
2 22.32± 0.4aA 24.32± 2.40aA 26.08± 1.68aB
3 21.31± 0.4aA 24.82± 0.48bA 27.83± 0.48cAB
4 21.76± 0.5aA 20.63± 0.59aC 19.87± 0.44aC
5 31.84± 0.4aB 31.89± 0.51aB 31.85± 0.53aA
4. Cu (µmol/l) 1 9.46± 4.13aA 9.71± 2.90aA 9.56± 1.73aA
2 10.51± 3.6aA 11.81±1.97aA 12.00± 1.38aA
3 9.44± 1.35aA 10.43± 1.57aA 10.70±3.12aA
4 9.78± 2.29aA 9.67± 1.92aA 9.58± 2.13aA
5 12.35± 0.1aA 12.38± 0.13aA 12.40± 0.09aA

Values with different superscript (small letters in rows and capital letters in columns) differ significantly (p< 0.05)

Conclusion

Efficacy of treatment of fasciolosis in bovine treated with triclabendazole and closantel showed significant (p<0.05) increase in TPP, albumin, A: G ratio, AST, Ca and Pi levels at 21st day of treatment whereas, Artemisia maritima treated group showed non-significant improvement. So the present study concluded that antifasciola efficacy of closantal and triclabendazole based on biochemical and mineral profile in bovines affected with fasciolosis is higher when compared with Artemisia maritima.

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