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, 14^th and 21^st 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.)

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)

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)

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 21^st 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.

References

1. Abouzeid NZ, Selim AM and El-Hady KM. 2010. Prevalence of gastrointestinal parasites infections in sheep in the Zoo garden and Sinai district and study the efficacy of anthelmintic drugs in the treatment of these parasites. Journal of American Science, 6 (11): 12-14.
2. Al-Saffar TM. 2008. Some haematological changes in sheep with chronic fascioliasis in Mosul. AL-Qadisiya Journal of Veterinary Medicine Science, 7 (1): 34-36.
3. Brennan GP, Fairweather I, Trudgett A, Hoey E, Mccoy M, Mcconville M, Meaney M, Robinson M, Mcferran N, Ryan L, Lanusse C, Mottier L, Alvarez L, Solana H, Virkel G, Brophy PM. 2007. Understanding triclabendazole resistance. Experimental and Molecular Pathology. 82:104-109.
4. Chakraborty D. 1999. Haemato-biochemical changes in Fasciola gigantica infected calves. Indian Journal of Animal Health, 36(1): 87-88.
5. Edith R, Sharma RL, Godara R and Thilagar MB. 2012. Experimental studies on anaemia in Riverine buffaloes (Bubalus bubalis) infected with Fasciola gigantica. Comparative Clinical Pathology, 21 (4): 415-419.
6. El-Khadrawy HH, Faragalla M, Moghazy El, Aziz, MM and Ahmed WM. 2008. Field investigation on the correlation between ovarian activity and fascioliosis in buffalo-cows. American-Eurasian Journal of Agriculture & Environment Sciences, 3(4): 539-546.
7. Lotfollahzadeh S, Mohri M, Bahadori SR, Dezfouly MR and Tajik P. 2008. The relationship between normocytic, hypochromic anaemia together with hepatic enzyme activities in cattle affected with Fasciola hepatica. Journal of Helminthology, 82: 85-88.
8. Pal S and Dasgupta C. K. 2006. Haemato-Biochemical Profiles of Buffalo In Anthelmintics Treatment Against Fasciola gigantica Infection. Buffalo Bulletin 25(2): 40.
9. Radostits OM, Gay CC, Blood DC and Hinchcliff KW. 2007. A Textbook of the Diseases of Cattle, Sheep, Pigs, Goats and Horses. 9^th ed. pp 1378-1382. W. B. Saunders Company, Philadelphia.
10. Rondelaud D, Vignoles P, Abrous M and Dreyfuss G. 2001. The definitive and intermediate hosts of Fasciola hepatica in the natural watercress beds in central France. Parasitology Research, 87: 475-478
11. Schweizer G, Braun U, Deplazes P and Torgerson PR. 2005. Estimating the financial losses due to bovine fasciolosis in Switzerland. Veterinary Record, 157: 188–193.
12. Sheikh GN, Qadri SGJ, Willayat MM and Gunjan D. 2006. Biochemical Profile of cattle naturally infected with Fasciola gigantica and F. hepatica. Journal of Veterinary Parasitology, 20(1): 41-43.
13. Snedecor GW. and Cochran WG. 1994. Statistical Methods. Viii edn. Oxford and IBH Publishing Company, New Delhi.
14. Soulsby EJL. 1987. Parasitologiay Enfermedades Parasitarias, seventh edition. Intermericana, Mexico D. F., Mexico, 40: 235–236.
15. Torgerson P and Claxton J. 1999. Epidemiology and Control, Dalton, J. P. Editor, Fasciolosis, CAB International, Wallingford, 113–149.