Parasitic diseases remain a major threat to livestock production around the world, particularly in India. Paramphistomosis caused by paramphistomes are one of the most common and economically important diseases of livestock. The high incidence of resistance to chemotherapeutics, toxicity and side effects has urged the necessity of finding alternative plant-based anthelmintics against helminth parasites. Therefore, the present investigation was aimed to assess the anthelmintic effect of the rind of Punica granatum Ethanol Extract (PgEE) against paramphistomes in infected sheep. Infected sheep were treated orally with 30 and 50 mg/ml concentrations of PgEE. Eggs Per Gram (EPG) count on faeces, haematological and biochemical parameters of sheep were investigated. In PgEE-treated sheep, the egg count reduced significantly in the faeces and the reduction was proportional to dosage and duration after treatment. The maximum reduction (97.95 %) was observed on day 21 post-treatment with 50 mg/ml concentration of PgEE. In infected sheep, the haemoglobin and protein content were below standard physiological values. Improvement of haematobiochemical profile was observed in sheep after treatment with PgEE.
Article Citation:
Lalhmingchhuanmawii K, Veerakumari L and Raman M.
Anthelmintic activity of Punica granatum ethanol extract against paramphistomes in infected sheep.
Journal of Research in Animal Sciences (2014) 2(1): 079-086.
Full Text:
http://janimalsciences.com/documents/AS0025.pdf
Título original
Anthelmintic Activity of Punica Granatum Ethanol Extract Against paramphistomes in infected sheep
Parasitic diseases remain a major threat to livestock production around the world, particularly in India. Paramphistomosis caused by paramphistomes are one of the most common and economically important diseases of livestock. The high incidence of resistance to chemotherapeutics, toxicity and side effects has urged the necessity of finding alternative plant-based anthelmintics against helminth parasites. Therefore, the present investigation was aimed to assess the anthelmintic effect of the rind of Punica granatum Ethanol Extract (PgEE) against paramphistomes in infected sheep. Infected sheep were treated orally with 30 and 50 mg/ml concentrations of PgEE. Eggs Per Gram (EPG) count on faeces, haematological and biochemical parameters of sheep were investigated. In PgEE-treated sheep, the egg count reduced significantly in the faeces and the reduction was proportional to dosage and duration after treatment. The maximum reduction (97.95 %) was observed on day 21 post-treatment with 50 mg/ml concentration of PgEE. In infected sheep, the haemoglobin and protein content were below standard physiological values. Improvement of haematobiochemical profile was observed in sheep after treatment with PgEE.
Article Citation:
Lalhmingchhuanmawii K, Veerakumari L and Raman M.
Anthelmintic activity of Punica granatum ethanol extract against paramphistomes in infected sheep.
Journal of Research in Animal Sciences (2014) 2(1): 079-086.
Full Text:
http://janimalsciences.com/documents/AS0025.pdf
Parasitic diseases remain a major threat to livestock production around the world, particularly in India. Paramphistomosis caused by paramphistomes are one of the most common and economically important diseases of livestock. The high incidence of resistance to chemotherapeutics, toxicity and side effects has urged the necessity of finding alternative plant-based anthelmintics against helminth parasites. Therefore, the present investigation was aimed to assess the anthelmintic effect of the rind of Punica granatum Ethanol Extract (PgEE) against paramphistomes in infected sheep. Infected sheep were treated orally with 30 and 50 mg/ml concentrations of PgEE. Eggs Per Gram (EPG) count on faeces, haematological and biochemical parameters of sheep were investigated. In PgEE-treated sheep, the egg count reduced significantly in the faeces and the reduction was proportional to dosage and duration after treatment. The maximum reduction (97.95 %) was observed on day 21 post-treatment with 50 mg/ml concentration of PgEE. In infected sheep, the haemoglobin and protein content were below standard physiological values. Improvement of haematobiochemical profile was observed in sheep after treatment with PgEE.
Article Citation:
Lalhmingchhuanmawii K, Veerakumari L and Raman M.
Anthelmintic activity of Punica granatum ethanol extract against paramphistomes in infected sheep.
Journal of Research in Animal Sciences (2014) 2(1): 079-086.
Full Text:
http://janimalsciences.com/documents/AS0025.pdf
Anthelmintic activity of Punica granatum ethanol extract against
paramphistomes in infected sheep
Keywords: Punica granatum, paramphistomosis, EPG, FECR % 079-086 | JRB | 2014 | Vol 2 | No 1 This article is governed by the Creative Commons Attribution License (http://creativecommons.org/ licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited. www.janimalsciences.com Journal of Research in Animal Sciences An International Scientific Research Journal Authors: Lalhmingchhuanmawii K 1 , Veerakumari L 1* and Raman M 2 .
Institution: 1. PG and Research Department of Zoology, Pachaiyappas College, Chennai - 600 030, Tamil Nadu, India.
2. Programme Director (Vaccine), Translational Research Platform for Veterinary Biologicals, TANUVAS, Chennai 600 007, Tamil Nadu, India.
Corresponding author: Veerakumari L.
Email:
Web Address: http://janimalsciences.com/ documents/AS0025.pdf. Dates: Received: 06 Feb 2014 Accepted: 25 Feb 2014 Published: 14 Mar 2014 Article Citation: Lalhmingchhuanmawii K, Veerakumari L and Raman M. Anthelmintic activity of Punica granatum ethanol extract against paramphistomes in infected sheep. Journal of Research in Animal Sciences (2014) 2(1): 079-086 An International Scientific Research Journal Original Research Journal of Research in Animal Sciences J o u r n a l
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ABSTRACT:
Parasitic diseases remain a major threat to livestock production around the world, particularly in India. Paramphistomosis caused by paramphistomes are one of the most common and economically important diseases of livestock. The high incidence of resistance to chemotherapeutics, toxicity and side effects has urged the necessity of finding alternative plant-based anthelmintics against helminth parasites. Therefore, the present investigation was aimed to assess the anthelmintic effect of the rind of Punica granatum Ethanol Extract (PgEE) against paramphistomes in infected sheep. Infected sheep were treated orally with 30 and 50 mg/ml concentrations of PgEE. Eggs Per Gram (EPG) count on faeces, haematological and biochemical parameters of sheep were investigated. In PgEE-treated sheep, the egg count reduced significantly in the faeces and the reduction was proportional to dosage and duration after treatment. The maximum reduction (97.95 %) was observed on day 21 post-treatment with 50 mg/ml concentration of PgEE. In infected sheep, the haemoglobin and protein content were below standard physiological values. Improvement of haematobiochemical profile was observed in sheep after treatment with PgEE.
INTRODUCTION Gastrointestinal helminth infection is one of the common and economically important diseases of grazing livestock (Perry et al., 2002). The clinical symptoms of GI helminth infections are decreased appetite, weight loss, reduction of meat and milk production, foul- smelling diarrhoea and dehydration (Githigia et al., 2005). Constant competition for the nutrients and tissue damage caused by the immature flukes during feeding and migration leads to oedema, anorexia and anaemia which ultimately results in the morbidity and mortality of host animals particularly in young, aged and immunosuppressed animals (Eysker and Pleoger, 2003). Among helminth parasites, paramphistomes are the most common and pathogenic found in the rumen and reticulum of sheep, goats, cattle and buffaloes (Manna et al., 1994). In domestic ruminants, high mortality rate is due to immature amphistomes (Hassan et al., 2005). Synthetic anthelmintics are used to control helminth infections in livestock. However, all sheep and goat farming countries reported anthelmintic resistance (Jackson and Coop, 2000; Sangster and Dobson, 2002; Kaplan, 2004). The resistance to commercially available anthelmintic drugs and the entry of drug residues in the food chain have instigated the search for alternative drugs such as medicinal plants. Medicinal plants have its origin in ethnoveterinary medicine and have been used for the prevention and treatment of gastrointestinal parasitism in many parts of the world (Athanasiadou et al., 2007). Punica granatum commonly known as pomegranate has been the focus of classical reviews for more than 100 years (Li et al., 2002) and various parts of the bark, leaves, immature fruits, and fruit rind have some therapeutic properties. In Ayurvedic medicine, the bark and root have shown to possess anthelmintic and vermifuge properties (Wang et al., 2010). Active constituents isolated from the stem bark are the alkaloids viz. pelletierine, isopelletierine, pseudopelletierine and methyl isopelletierine (Neuhofer et al., 1993). An experiment on a living subject is more preferable because it provides more insight in the aspects of the studies, therefore, in vivo studies is frequently employed over in vitro studies. The in vivo assay such as the faecal egg count reduction test is suitable for the evaluation of all types of anthelmintics (Verma et al., 2006). Furthermore, in vivo observations on the haematological and biochemical profiles of the animals can offer more insights into the safety and protective efficacy of the plant extracts. Haematological parameters such as haemoglobin (Hb), packed cell volume (PCV), total erythrocyte count (TEC), total leucocyte count (TLC) and differential count (DC) are important diagnostic tools for assessing healthiness of host animals. Biochemical profiles such as serum glucose, total serum protein (TSP), albumin (A), globulin (G), aspartate aminotransferase (AST) and alanine amino transferase (ALT) are likely to be affected because of infection and therefore may exhibit quantitative changes prior to and after the treatment (Priya et al., 2013). In view of the above, the present study was undertaken to assess the efficacy of Punica granatum Ethanol Extract against paramphistomes in naturally infected sheep based on the changes in the number of paramphistome eggs in the faeces, haematological and biochemical profiles.
MATERIALS AND METHODS The present study was undertaken on a sheep farm in Thiruthavalli, 30 km south of Chennai, Tamil Nadu, India. A total of 200 sheep (Madras Red) were randomly selected for the study. The study animals were put out to graze in a fenced area within the farm premises. Preparation of Punica granatum ethanol extract About 2 kg of the rind of Punica granatum were cleaned, shade dried and coarsely powdered. Successive solvent extraction was done by cold percolation method Lalhmingchhuanmawii et al., 2014 080 Journal of Research in Animal Sciences (2014) 2(1): 079-086 by soaking in hexane, chloroform, ethyl acetate and ethanol successively in an aspirator bottle for 48 h. After 48 h, they were filtered by Whatman Filter paper No.1. The solvent was removed by distillation using Evator Rotary Evaporator and the extracts were concentrated and dried in Lyodel Freeze Dryer. The stock solution (100 mg/ml) of PgEE was then serially diluted with water to obtain 30 and 50 mg/ ml concentrations. Experimental animals The experimental trial was conducted in naturally infected sheep. Out of 200 sheep, twenty four sheep of either sex aged six months and above naturally infected with paramphistomes were selected based on egg counts. Animals having faecal egg count more than 500 eggs per gram were selected and divided into four groups of six animals each. Experimental design Twenty four naturally paramphistome infected sheep were divided into four groups - group I, group II, group III and group IV. Group I animals were dewormed prior to the experimental period with albendazole (at the manufacturer-recommended dose) served as uninfected (healthy) control. Group II served as infected-untreated control. Group III and IV were treated orally with PgEE at two different concentrations, 30 and 50 mg/ml respectively on day 0. Faecal examination Faecal samples for examination were collected per rectum at day 0 (before treatment) and days 7, 14 and 21 post treatment. The McMasters method was employed for the quantitative estimation of eggs per gram (Soulsby, 1982). The percentage reduction in faecal egg count (FECR %) in groups III and IV were calculated. Haematological and biochemical analysis Blood samples were drawn from the jugular veins with the help of a sterilized dry syringe and transferred to two disposable blood collecting tubes, BD Vacutainer Sodium Heparin 68 USP units and BD Vacutainer Serum on day 0 before treatment and on days 7 and 21 post treatment from all the four groups. Blood from BD Vacutainer Sodium Heparin 68 USP unit was used for the evaluation of haematological studies such as Hb, PCV, TEC, TLC, neutrophils (N), lymphocytes (L) and eosinophils (E) (Jain, 1986). Blood from BD Vacutainer Serum was allowed to coagulate at 37 C for 30 min. The serum was separated by centrifugation at 3000 rpm for 10 min and was used for the biochemical assay. Serum samples were analysed for TSP, albumin, globulin, A/G ratio, glucose, AST and ALT. Statistical analysis All the data obtained in the present study were statistically analysed using the statistical software SPSS version 16.0. One-way Anova using Bonferroni test was applied to find out the significant difference between healthy, infected-untreated and PgEE-treated sheep.
RESULTS AND DISCUSSION The mean EPG and FECR % in healthy, infected -untreated and PgEE-treated sheep were given in Table 1. Throughout the study period, the faeces of healthy sheep (Group I) contain no eggs of paramphistomes. In contrast, faeces of infected-untreated (Group II) showed absolutely high counts during the whole period of observation. In PgEE-treated sheep (Group III and IV), the egg counts reduced significantly on day 7 post- treatment and continued to decline further till day 21 post -treatment. The maximum reduction was recorded as 97.95 % happening on day 21 after treatment with 50 mg/ml of PgEE. The EPG counts and FECR % on days 0, 7, 14 and 21 are significantly different (P < 0.01). The statistical analysis (Bonferroni test) of Table 1 showed that the values of all the four groups (I-IV) differ significantly on days 0, 7, 14 and 21 (P <0.01). The values of group II on the different days of observations (0, 7, 14 and 21) do not differ significantly (P > 0.05). There is significant difference (P <0.05) Lalhmingchhuanmawii et al., 2014 Journal of Research in Animal Sciences (2014) 2(1): 079-086 081
between values of group III and IV on all days of observations. There is no significant difference between the egg counts of all the three groups (II-IV) on day 0. The results of the haematological indices were given in Table 2. The Hb, PCV, TEC and TLC in healthy sheep (Group I) were in a range of 12-13 g %, 33-34 %, 910 5 /cmm and 9-1110 3 /cmm respectively throughout the period of investigation whereas the Hb, PCV and TEC level of the infected sheep was lesser than the healthy sheep. High level of TLC was observed in infected sheep. Treatment with 30 and 50 mg/ml concentrations of PgEE gradually improved the Hb, PCV and TEC to normal range and the improvement level was dose and time dependent. The TLC level was also brought down to a count comparable to normal range after treatment with 30 and 50 mg/ml concentrations of PgEE on 21 days PT. Significant reduction of neutrophil and eosinophil was observed after treatment with PgEE. Infected sheep treated with 30 and 50 mg/ml concentrations of PgEE increased the lymphocyte level in groups III and IV gradually. The statistical analysis (Bonferroni test) of Table 2 revealed that the values of all the four groups (I-IV) differ significantly on days 0, 7 and 21 (P < 0.01). There is significant difference (P < 0.05) between values of group III and group IV on all days of observations. The values of group II on the different days of observations (0, 7 and 21) do not differ significantly (P >0.05). There is no significant difference (ns) from the respective group I values (P > 0.05). The observations on the biochemical profiles of paramphistomes infected sheep before and after treatment with 30 and 50 mg/ml concentrations of PgEE are presented in Table 3. The TSP level of healthy sheep (Group I) was approximately 7 g/dl throughout the period of investigation whereas the TSP level of the infected-untreated sheep (Group II) was significantly less, about 4 g/dl. The TSP level of the infected sheep significantly increases following treatment with PgEE. The albumin, globulin and A/G ratio of infected sheep was found to be lower than the healthy sheep, but showed a similar trend of increase following treatment with PgEE. Treatment of infected sheep with 30 and 50 mg/ml concentrations of PgEE had no significant effect on the glucose level. High levels of AST and ALT were observed in the infected sheep but the levels returned to normal after treatment with PgEE. Lalhmingchhuanmawii et al., 2014 082 Journal of Research in Animal Sciences (2014) 2(1): 079-086 Groups Pre-treatment (0 day) Days post-treatment EPG count FECR % 7 14 21 0 7 14 21 I 0 0.00 0 0.00 0 0.00 0 0.00 - - - - II 564 34.29 552 32.79 564 30.75 576 25.17 - - - - III 576 48.00 144 15.17 120 09.79 72 05.36 - 75.00 79.16 87.50 IV 588 27.79 48 09.79 24 06.19 12 04.89 - 91.83 95.91 97.95 Table 1 EPG count and FECR % in healthy (Group I), infected-untreated (Group II) and PgEE-treated (Groups III and IV) sheep Each value represent mean SD of n = 6 Group I - Healthy sheep Group II - Infected-untreated sheep Group III - Infected sheep treated with 30 mg/ml PgEE Group IV - Infected sheep treated with 50 mg/ml PgEE The statistical analysis (Bonferroni test) of Table 3 exposed that the values of all the four groups (I-IV) differ significantly on days 0, 7 and 21 (P < 0.01) except for glucose content which does not show any significant difference (P > 0.05). There is significant difference (P < 0.05) between values of group III and group IV on all days of observations. The values of group II on the different days of observations (0, 7 and 21) do not differ significantly (P > 0.05). There is no significant difference (ns) from the respective group I values (P > 0.05). In the present investigation, the EPG count of paramphistome eggs were significantly reduced following oral administration of PgEE. Priya et al., Lalhmingchhuanmawii et al., 2014 Parameters Days Groups I II III IV Hb (g%) 0 13.02 0.81 07.48 0.33 07.78 0.54 07.12 0.39 7 12.59 0.47 07.15 1.03 08.41 1.19 ns 10.22 0.43 21 12.68 0.50 07.41 0.75 09.72 1.21 ns 12.69 0.93 ns
21 02.30 0.35 05.90 0.25 02.80 0.20 01.70 0.17 Table 2 Haematological parameters of healthy (Group I), infected-untreated (Group II) and PgEE-treated (Groups III and IV) sheep Each value represent mean SD of n = 6 Group I - Healthy sheep Group II - Infected-untreated sheep Group III - Infected sheep treated with 30 mg/ml PgEE Group IV - Infected sheep treated with 50 mg/ml PgEE
Standard physiological values Hb (8-16 g%); PCV (24-40 %); TEC (8-15 10 5 /cmm); TLC (4-12 10 5 /cmm); N (25-35 %); L (60-70 %); E (1-3 %) Journal of Research in Animal Sciences (2014) 2(1): 079-086 083
(2013) reported a similar reduction in faecal egg count of sheep infected with paramphistomes after treatment with aqueous extract of pods of Acacia concinna. Faecal egg count reduction of gastrointestinal nematodosis in Garole sheep treated with ivermectin, levamisole and albendazole was also reported by Pandit et al., (2009). The present findings are further supported by the studies of Verma et al., (2006) and Diaz et al., (2006), who Lalhmingchhuanmawii et al., 2014 Parameters Days Groups I II III IV Total serum protein (g/dl) 0 07.12 00.55 04.87 00.39 05.14 00.21 04.74 00.40 7 07.25 00.53 04.49 00.24 06.39 00.34 06.84 00.32 21 07.54 00.25 04.84 00.32 07.27 00.31 07.65 00.27 Albumin (g/dl) 0 03.03 00.62 01.03 00.30 01.23 00.28 01.17 00.38 7 03.16 00.32 01.28 00.31 03.16 00.43 ns 03.25 00.40 ns
Table 3 Biochemical profile of healthy (Group I), infected-untreated (Group II) and PgEE-treated (Groups III and IV) sheep Each value represent mean SD of n = 6 Group I - Healthy sheep Group II - Infected-untreated sheep Group III - Infected sheep treated with 30 mg/ml PgEE Group IV - Infected sheep treated with 50 mg/ml PgEE
Standard physiological values Total serum protein (6-7.9 g/dl); Albumin (2.4-3.9 g/dl); Globulin (3.5-5.7 g/dl); A/G ratio (1-1.2); Glucose (40-80 g/dl); AST (98-278 U/L); ALT (24-83 U/L) 084 Journal of Research in Animal Sciences (2014) 2(1): 079-086 reported a significant reduction in egg count in buffaloes and cattle naturally infected with paramphistomes after treatment with chemical anthelmintics. A decrease in Hb, PCV, TEC, lymphocytes, TSP, albumin, globulin, A/G ratio and increase in TLC, neutrophils, eosinophils, AST and ALT levels were observed in the infected sheep prior treatment. The fall in Hb, PCV and TEC probably implies that the animals were suffering from anaemia, due to the haemorrhage caused by the immature parasites. Verma et al., (2006) and Diaz et al., (2006) also reported a decrease in Hb, PCV and TEC in cattle naturally infected with paramphistomes. A lower value of TSP, albumin and globulin and high values of AST and ALT in cattle and buffaloes infected with paramphistomes were also reported by Bharti and Prasad (2001). The fall in the protein level observed in the present study might be due to haemodilution, a compensatory mechanism for intestinal haemorrhage caused by migrating flukes and later on due to loss of large quantities of serum protein in the gut through exudation. Hypoproteinemia in helminthosis is primarily the result of hypoalbuminaemia. Hypoproteinemia, coupled with loss of appetite, seems to be the most important pathophysiological consequence of paramphistomosis (Priya et al., 2013). The changes in serum biochemical constituents during Fasciola/ Paramphistomum infection reflect disturbances in liver function caused due to tissue damage and fluid loss caused by the parasites in situ. Recovery from the damages of this vital organ following treatment of the infected animals may result in the resumption of liver functions and restoring the body fluid balance (Bharti and Prasad, 2001). The levels of AST and ALT reduced after treatment with 30 and 50 mg/ml concentrations of PgEE and the decline in AST and ALT level was proportional to dosage and duration. A similar result of the decline in AST and ALT was reported by Bharti and Prasad (2001) in cattle following treatment with oxyclozanide. Due to PgEE treatment, the levels of Hb, PCV, TEC, TLC, neutrophils, lymphocytes, eosinophils, TSP, albumin, globulin, A/G ratio, AST and ALT approached normal values in paramphistome infected sheep. This study clearly revealed the anthelmintic efficacy of PgEE against paramphistomes. Similar findings on the improvement in haematological and biochemical profiles of treated animals using synthetic anthelmintic drugs have been reported by several researchers (Bharti and Prasad, 2001; Diaz et al., 2006; Verma et al., 2006). Priya et al., (2013) also reported a similar outcome on the haematological and biochemical profiles of the sheep infected with paramphistomes following treatment with aqueous extract of pods of Acacia concinna.
CONCLUSION Observations, in the present study, on EPG count and haemato-biochemical parameters among healthy, infected-untreated and PgEE-treated sheep revealed that PgEE is highly effective against paramphistomes causing an improvement in the health status of the infected host. Therefore, PgEE could be successfully used as an anthelmintic to treat paramphistomosis in livestock.
REFERENCES Athanasiadou S, Githiori J and Kyriazakis I. 2007. Medicinal plants for helminth parasite control: facts and fiction. Animal. 1(9): 1392-1400.
Bharti P and Prasad KD. 2001. Biochemical profiles of cattle and buffalo infected with Paramphistomum spp. and Fasciola gigantica. J. Vet. Parasitol., 15(2):149-151.
Diaz P, Lomba C, Pedreira J, Arias M, Snchez- Andrade R, Surez JL, Dez-Baos P, Morrondo P and Paz-Silva A. 2006. Analysis of the IgG antibody response against Paramphistomidae trematoda in naturally infected cattle: Application to serological Journal of Research in Animal Sciences (2014) 2(1): 079-086 085 Lalhmingchhuanmawii et al., 2014 surveys. Vet. Parasitol., 140(3-4): 281-288.
Eysker M and Ploeger HW. 2003. Value of present diagnostic methods for gastrointestinal tract nematodes infection in ruminant. In: Symposia of the British Society for Parasitology, Cambridge University Press, UK. 37: 109-119.
Githigia SM, Thamsborg SM, Maingi N and Munyua WK. 2005. The epidemiology of gastrointestinal nematodes in goats in the low potential areas of Thika district, Kenya. Bull. Anim. Health Prod Afr., 53(1):5-12.
Hassan SS, Kaur K, Joshi K and Juyal PD. 2005. Epidemiology of paramphistomosis in domestic ruminants in different districts of Punjab and other adjoining areas. J. Vet. Parasitol., 19(1): 43-46.
Jackson F and Coop RL. 2000. Development of anthelmintic resistance in sheep nematodes. Parasitol., 120(7): 95-107.
Jain NC. 1986. Hematological techniques. In: Schalms Veterinary Hematology. Lea and Febijer Publishers, Philadelphia, USA. 20-86.
Kaplan RM. 2004. Drug resistance in nematodes of veterinary importance: a status report. Trends in Parasitology. 20(10): 477-481.
Li HX, Wang Z, Liu YZ. 2002. Progress in studies on chemical constituents and pharmacological effects of Punicaceae. Chinese Traditional and Herbal Drugs. 33: 765-769.
Manna AK, Pramanik S and Mukherjee GS. 1994. Incidence of paramphistomiasis in West Bengal. Indian J. Anim. Hlth., 33(2): 87-89.
Neuhofer H, Witte L, Gorunovic M and Czygan FC. 1993. Alkaloids in the bark of Punica granatum L. (pomegranate) from Yugoslavia. Pharmazie. 48(5): 389- 391. Pandit S, Ghosh JD, Chinya A, Mandal R, Jas R and Moi S. 2009. Evaluation of anthelmintic efficacy of ivermectin, levamisole and albendazole against naturally occurring gastrointestinal nematodosis in Garole sheep. J. Vet. Parasitol., 23(2): 121-125.
Perry BD, Randolph TF, Mc Dermott JJ, Sones KR and Thornton PK. 2002. Investing in animal health research to alleviate poverty. ILRI (International Livestock Research Institute), Nairobi, Kenya. 148.
Priya P, Veerakumari L and Raman M. 2013. Anthelmintic efficacy of Acacia concinna against paramphistomes in naturally infected sheep. J. App. Anim. Res., 41(2): 183-188.
Sangster NC and Dobson RJ. 2002. Anthelmintic resistance. In: The biology of nematodes. Editor D. Lee, Taylor and Francis. 531-567.
Soulsby EJL. 1982. Helminths, Arthropods and Protozoa of domesticated Animals. 7th edition, London, UK: Blackwell Scientific Publications.
Verma SK, Gupta MP and Juyal PD. 2006. Efficacy of oxyclozanide and triclabendazole in buffaloes infected with paramphistomosis. J. Vet. Parasitol., 20(1): 61-64.
Wang R, Ding Y, Liu R, Xiang L and Du L. 2010. Pomegranate: Constituents, bioactivities and pharmacokinetics. Fruit, vegetable and cereal science and biotechnology. 4(2): 77-87. 086 Journal of Research in Animal Sciences (2014) 2(1): 079-086 Lalhmingchhuanmawii et al., 2014 Submit your articles online at janimalsciences.com
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