group as compared to MNB supplemented (T 2

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1 Indian J. Anim. Res., 50 (5) 2016 : Print ISSN: / Online ISSN: AGRICULTURAL RESEARCH COMMUNICATION CENTRE Effect of tanniferous leaf meal mixture based multi nutrient blocks on nutrient utilization and biochemical profile of Haemonchus contortus infected goats Surender Singh, A.K. Pathak*, Muzaffer Khan and R.K. Sharma Division of Animal Nutrition, Faculty of Veterinary Science & AH, SKUAST-Jammu , India. Received: Accepted: DOI: /ijar.5716 ABSTRACT The experiment was carried out in 12 local adult male goats of average body weights (26.49±0.87, kg) and they were allocated in 3 groups (4 goats in each). Multi nutrient blocks (MNB) were supplemented in first two groups i.e. in T 1 (no infection) and T 2 (Haemonchus contortus 1500 larvae) group, while, condensed tannins (CT) containing multi nutrient blocks (MNB-CT) were supplemented in T 3 (H. contortus 1500 larvae) group in completely randomized block design during a feeding trial of 75 days. All the animals were fed concentrate 100g/ day, adlib wheat straw and MNB or MNB-CT to meet their maintenance requirement. Daily intakes (kg W 0.75 ) of dry matter (DM), organic matter (OM), concentrate mixture, MNB, MNB-CT, wheat straw and digestibility of DM, OM, crude protein (CP) and acid detergent fibre (ADF) differed significantly (P<0.05) irrespective of groups. MNB-CT supplementation significantly (P<0.001) reduced serum urea (mg/dl) level and faecal egg counts (FECs) in T 3 group as compared to MNB supplemented (T 2 ) group. Serum proteins differed significantly (P<0.001) among groups. It may be concluded that MNB-CT supplementation indicates a positive impact on protein bioavailability and encouraging impact on nutrient utilization, biochemical constituents and health status of goats by lowering serum urea and FECs. Key words: Condensed tannins, Goats, Haemonchus contortus, Leaf meal mixture, Multi nutrient blocks, Nutrient utilization. INTRODUCTION Goats contribute greatly to the agrarian economy, especially in areas where crop and dairy farming are not economical. Goat farming is crucially important for provision of milk, meat, hides and manures for human need and also as a source of income for many poor farmers and landless labours at village levels in developing countries. Gastrointestinal nematodes (GINs) remain one of the major threats to sheep and goats in all over the world. Among the GINs, Haemonchus contortus is one of the most pathogenic GINs, which inhabits the abomasum, suck blood from the host and results in anaemia (Rowe et al., 1988). The adverse effect on productive performance of sheep and goats is manifested in a variety of ways. Changes in body weight vary with the level of infection, the species of GINs, and age of the animal, nutritional and immunological status of the host are the most common features of GIN infections (Anderson, 1982; Pathak, 2011, 2013; Pathak et al., 2013a). They impair animal productivity through reductions in voluntary feed intake and / or reductions in the efficiency of feed utilization, particularly by inefficient absorption of nutrients in the gastrointestinal tract (Coop and Kyriazakis, 2001; Pathak and Tiwari, 2012). This is most pronounced for protein, for which a reduced absorption and / or retention in parasitized animals has repeatedly been shown (Pathak and Tiwari, 2013a; Pathak et al., 2013a). The magnitude of these effects is influenced by the size of larval challenge, number and species of GIN present and the nutritional status of affected animals (Pathak et al., 2013a). Control of these GINs has been based on the repeated use of synthetic anthelmintics. However, the development of GINs resistant to synthetic anthelmintics currently available (Torres-Acosta and Hoste, 2008) and concerns regarding anthelmintic residues in food chain and in the environment have stimulated the search for alternative sustainable non-chemical GINs control strategies (Athanasiadou et al., 2008; Pathak, 2013; Pathak et al., 2013a). An increasing number of recent studies indicates that nutrition could affect parasitism not only through quantitative variations of different diet components but also by the presence of some qualitative compounds in plants consumed by ruminant, and particularly condensed tannins (Athanasiadou et al., 2003; Pathak et al., 2014). Two main hypotheses have been proposed to explain the mechanism of action of CT on the GINs. The CT might interfere directly with the biology of various developmental stages of GINs *Corresponding author s dranandpathak1974@gmail.com

2 726 INDIAN JOURNAL OF ANIMAL RESEARCH (Pathak et al., 2013 b,c). On the other hand, indirectly, CT could also improve the host nutrition by protecting the dietary proteins from ruminal degradations which could in turn worm biology. The information on the supplementation of CT through LMM containing multi-nutrient blocks (MNB-CT) in animals infected with GINs is scanty. Therefore, the CT source from LMM to be used in MNB-CT to reduce H. contortus in goats warrants investigation. Keeping all these points in view the present study has been taken to investigate the effect of MNB-CT supplementation on nutrient intake and utilization, biochemical profile, health status and performance of H. contortus infected goats. MATERIALS AND METHODS The experimental study was carried out at Division of Animal Nutrition, Faculty of Veterinary Sciences and Animal Husbandry, Sher-e-Kashmir University of Agricultural Sciences and Technology R. S. Pura, Jammu, India. Experimental animals, feeds and design: Twelve local adult male goats (26.49 ± 0.87 kg average body weight) were divided randomly into three equal groups (4 goats in each) in a completely randomized block design (CRD). Basal diet of wheat straw Ad libitum and concentrate mixture (100g / goat) were offered to goats of all three groups. Concentrate mixture was formulated from locally available feed ingredients (maize: 30, wheat bran: 22, mustardoil cake: 45, mineral mixture: 02 and common salt: 01, percent) for feeding of goats throughout the experiment. All experimental goats were fed as per NRC (2007) for maintenance. The MNB was offered ad libitum in T 1 (no infection) and in T 2 (H. contortus infection) groups, while, MNB-CT was offered ad libitum in T 3 (H. contortus infection) group. All goats were kept in a hygienic well-ventilated goat shed having concrete floor with the provision of individual feeding and watering. Before commencement of the experiment all goats were treated with different anthelmintics viz., tab. distodine 15 mg/kg body weight, orally and tab. panacur 5 mg/kg body weight orally alternated over a two weeks. The elimination of parasitic infection was confirmed by faecal examination. A feeding trial of 75 days duration was undertaken and a metabolism trial of 6 days collection period was conducted on all goats towards the end of feeding trial. Faeces voided and volume of urine excreted was recorded to determine the nutrient intake, digestibility and nitrogen (N) balance in goats. Samples and sampling procedures: The locally available tanniferous tree leaves were collected from faculty premises and transported to the divisional shed in fresh state. The tree leaves were shed dried and were processed for preparation of leaf meal mixture (LMM). The leaf meals were ground with electric grinder. The leaves of Eugenia jambolana and Psidium guajava were mixed in the ratio of 70: 30 for formulation of LMM. The infective 3 rd stage larvae (L 3 ) of H. contortus of caprine origin were produced by petridish method of faecal culture technique. The infective doses of L 3 larvae of H. contortus were prepared and administered 1500 L 3 per goat of T 2 groups. Blood samples were collected from all experimental animals at the start of the experiment (0 day) and thereafter at fortnightly intervals. About 4 ml of blood was collected aseptically in wide bore test tubes without anticoagulant from jugular vein of each goat for serum separation. The harvested serum was separated by centrifugation at 3500 x g for 5 minute and kept frozen at C. The faecal samples were collected at fortnightly intervals from all the goats directly from the rectum early in the morning before feeding and watering. The samples were processed and examined using Stoll s egg counting method. The representative samples of feeds, residue left and faeces voided were processed and ground to pass through 1 mm screen in Willey grinder. The samples of urine and faeces were collected in separate sample collection bags and the samples were processed and analysed as standard procedure. Preparation of multi nutrient blocks: The MNB and MNB- CT were prepared from locally available feed ingredients. The LMM was incorporated as CT source for preparation of MNB-CT blocks (Table 1). Both types of blocks were prepared by cold process as per standard method. The weighed amount of semi-solid mixture of 2.5 kg was put in manually operated multi nutrient block making machine and blocks were prepared by applying suitable pressure. Before feeding, the blocks were air dried in the shed for days, so as to be hard enough for handling, hanging and for licking purposes. Table 1: Ingredient composition (% DM) of MNB and MNB-CT Ingredient MNB MNB-CT Mustard oil cake Leaf meal mixture Molasses Urea Lime stone powder Di calcium phosphate Mineral mixture Wheat bran Common salt MNB: Multi nutrient block; MNB-CT: Condensed tannins containing multi nutrient block

3 Volume 50 Issue 5 (2016) 727 Chemical analysis: The CT was estimated by butanol-hcl method (Makkar, 2000). The processed samples of feed, residues and faeces were analyzed for proximate composition as per AOAC (1995) and fibre fractions (NDF and ADF) as per Van Soest et al. (1991). The nitrogen content in feed, residues, faeces, and urine was analyzed following Kjeldahl method (AOAC, 1995). Biochemical constituents were estimated by using readymade diagnostic kits. Glucose in serum was estimated by the glucose-oxidase peroxidise (GOD-POD) end-point assay as per Teitz (1976). Total serum protein (Tietz, 1986) and albumin (g/ dl) was done as per Doumas et al. (1972). Globulin was determined as the difference between total protein and albumin concentration in the serum. Serum urea (mg/ dl) level was estimated as per Tiffany et al. (1972). Aspartate amino transferase (AST) and alanine amino transferase (ALT) activities (IU/L) were determined by IFCC methods of Bergmeyer et al. (1986 a,b). Serum alkaline phosphatase (ALP; IU/L) was estimated as per the method described by Teitz (1976). Lactate dehydrogenase (LDH; IU/L) activity was determined by kinetic assay method as per Henry et al. (1960). Faecal egg counts (FECs) per gram of faeces were made using Stoll s egg counting method (Anonymous, 1986). Statistical analysis: The data obtained were subjected to analysis of variance and treatment means were ranked using Duncan s multiple range test (Snedecor and Cochran, 1994). The periodic alterations in biochemical profiles and parasitological parameters (FECs) were analyzed using repeated measures design (General linear model; GLM, Multivariate) by SPSS version 10.0 for windows. Significance was declared at P<0.05 unless otherwise stated. The FECs were zero in goats of T 1 group throughout the experiment, so they were not included in the statistical analysis. RESULTS AND DISCUSSION The ingredients and chemical composition (% DM) of feed (CM, MNB, MNB-CT and WS) offered to goats during experimental period are presented in Tables 1 and 2. The CT content of MNB-CT was 1.76 percent. The proportions and ingredient composition of multi nutrient blocks in the present study were contradictory to those used by earlier workers (Aye and Adegun, 2010; Mohammed et al., 2007; Somasiri et al., 2010). It depends upon the easy accessibility, availability and quality of local feed ingredients. However, the chemical composition of CM and WS used in the experiment was comparable with the values reported by many workers (Patra et al., 2006; Dey et al., 2008; Pathak et al., 2013a). Intake and digestibility of nutrients: Average body weight (kg) and metabolic body size (kg W 0.75 ) of goats differed significantly (P<0.05) among all three groups during metabolism trial. Daily nutrient intake by goats during the metabolism trial under three different groups is given in the Table 3. Daily intakes (g/kg W 0.75 ) of CM, WS, DM and OM were significantly higher (P<0.05) in T 2 group as compared to T 1 groups. However, multi nutrient block intake was significantly lower (P<0.05) in T 2 group as compared to T 1 groups. The intake (g/kg W 0.75 ) of multi nutrient block when compared between both infected groups (T 2 and T 3 ), significantly higher MNB-CT intake was recorded in T 3 than that of in T 2 group which might be attributable to the presence of CT. The present results are agreement with the findings of Kabasa et al. (2000) who reported that CT played a significant role in reducing the negative effects of GINs and suggested that goats prefer tanniferous diet to get rid of GIN load up to some extent. Daily nutrient intakes (g/kg W 0.75 ) in terms of DCP, DOM and TDN (g/kg W 0.75 ) were significantly higher in T 2 group as compared to T 1 groups which might be due to high intensity of H. contortus load in T 2 group. The findings suggested that nutrient intake and utilization was not affected adversely with CT supplementation in conformity with the previous reports (Dey et al., 2008; Pathak et al., 2013a).The DMI by goats was within the normal range (NRC, 2007) and this clearly indicates that MNB-CT blocks for experimental goats were palatable. Table 2: Chemical composition of feedstuffs (% DM) Attributes CM MNB MNB-CT WS Dry matter Organic matter Crude protein Crude fibre Ether extract Total Ash Acid insoluble ash Calcium Phosphorus Neutral detergent fibre Acid detergent fibre Condensed tannins CM: Concentrate mixture; WS: Wheat straw; MNB: Multi nutrient block; MNB-CT: Condensed tannins containing multi nutrient block

4 728 INDIAN JOURNAL OF ANIMAL RESEARCH Table 3: Effect of MNB and MNB-CT supplementation on intake and digestibility of nutrients and nitrogen balance in H. contortus infected goats Attributes Group* SEM P values T 1 T 2 T 3 Body weight (kg) b ± a ± ab ± Metabolic body size (kg w 0.75 ) b ± a ± b ± Feed intake (g/kg w 0.75 ) DM a ± b ± a ± OM a ± b ± a ± CM 7.76 a ± b ± a ± WS a ± b ± a ± MNB/ MNB-CT b ± a ± b ± CT intake (g/d) 0.00 ± ± ± Nutrient intake (g/kg w 0.75 ) DCP 3.25 a ± b ± a ± DOM a ± b ± ab ± TDN a ± b ± ab ± Nutrient digestibility (%) DM b ± a ± ab ± OM b ± a ± ab ± CP b ± a ± a ± EE ± ± ± NDF ± ± ± ADF b ± a ± ab ± Nitrogen balance (g/d) Intake 9.31 ± ± ± Faecal excretion 3.20 a ± b ± b ± Urinary excretion 3.55 b ± c ± a ± Retention 2.57 b ± a ± b ± ab Means with different superscripts within a row differ significantly; *T 1 : Negative control; T 2 : Infected control; T 3 : Infected Treatment, g/d: gram per day The palatability of blocks was closely related to be a threshold of CT content (low to moderate level) below which no adverse effect was evident on nutrient intake and utilization. The digestibility (%) of DM, OM, CP and ADF was significantly higher (P<0.05) in negative control (T 1 ) as compared to both infected groups (T 2 ), however, digestibilities of EE and NDF did not differ significantly (P<0.05) irrespective of groups. Present results are in concurrence with earlier reports, who have reported that CT are beneficial to ruminants at low concentration because they protect plant proteins from degradation in the rumen (Waghorn and Shelton, 1992; Wang et al., 1994; Dey et al., 2008; Pathak et al, 2013a). Higher CT level (more than 5%) in the diet of ruminants adversely affect the intake and digestibility of forage (Perevolotsky et al., 1993). Rowe et al. (1988) reported a reduction in apparent digestibility of OM across the whole digestive tract, but particularly in the abomasum of H. contortus infected sheep. However, the extent of reduction of course depends upon the worm burden (Pathak and Tiwari, 2012a) and the reduction is sometime small and often transitory in its nature. In the present study decreased digestibilities of DM, OM, CP and ADF in H. contortus infected goats are similar to earlier reports (Entrocasso et al., 1986; Pathak and Tiwari, 2012a; Pathak et al., 2013a) in infected kids, lambs and sheep (H. contortus) and calves (O. ostertagi). Present study revealed DCP and TDN intake was higher in T 3 group. The CT from MNB-CT improved the bioavailability of protein and reduced the detrimental effect caused by H. contortus by reducing the intensity of H. contortus infection in T 3 group. Nitrogen balance: The intake of N (g/d) did not differ significantly (P<0.05) among all three groups, however, N excreted (g/d) through faeces was significantly (P<0.001) lower in T 1 as compared to both infected groups (T 2 ) whereas urinary-n excretion (g/d) was found to be highest (P<0.004) in T 2 followed by T 1 and lowest urinary-n excretion in T 3 group. Nitrogen retained (g/d) were significantly (P<0.002) lower in T 2 as compared to T 3 and T 1 groups. Waghorn et al. (1994 a, b) also reported that a low to moderate level of CT in forage given to sheep has increased the nonammonia nitrogen flux to the small intestine, to increase the absorption of essential amino acids. Similar to the present study, increased N retention in sheep and goats given tanniferous feeds at low to moderate levels, lowered N excretion through urine has been reported by previous studies (Waghorn et al., 1994b; Scharenberg et al., 2008; Dey et al., 2008; Pathak et al., 2013a).

5 Volume 50 Issue 5 (2016) 729 Biochemical parameters: Various biochemical constituents and serum enzymes activity among all three groups are presented in Table 4. Significantly (P<0.05) lower glucose level was obtained in T 2 as compared to T 3 and T 1 group. In the present study, among both infected (T 2 ) groups the glucose level was significantly higher (P<0.015) in T 3 as compared to T 2, which might be due to high propionic acid concentration. Although, it was not measured in the present study but assumed to be higher due to MNB-CT supplementation because CT may act as propionate precursor. In the rumen, CT may decrease acetate to propionate ratio, resulting from an increased transfer of hydrogen to propionate (Dschaak et al., 2011). Since propionic acid is glucogenic in nature it might have converted in to glucose through the process of gluconeogenesis (Lehninger et al., 1993) and significantly higher glucose level was obtained in T 3 than that of T 2 group. As the period of H. contortus infection increased, serum glucose level declined significantly (P<0.001) in T 2 with no effect in T 3 group. This might be due to the effect of CT on diet and detrimental effect on H. contortus parasites. In the present study, significantly (P<0.001) lower serum urea was recorded in T 3 relative to T 1 and T 2 groups. Effects of CT supplementation were associated with low rumen ammonia concentration and rapid turnover of the plasma pool (Waghorn et al., 1994b). Serum urea level is an indicator of protein degradation in rumen. Significantly lower level of serum urea in T 3 group may be attributed to the reduced rumen protein breakdown and increased essential amino acid absorption (Waghorn et al., 1987; 1990). Similarly, lower serum urea concentration was Table 4: Effect of MNB-CT supplementation on biochemical profile and serum enzyme activity in H. contortus infected goats Attributes Period SEM P value 0 d 15 d 30 d 45 d 60 d 75 d G P G x P Glucose (mg/dl) T ca 56.5 ca bb abb abc ab T cb cb ba aba aba aa 1.81 T ca ca bb abb abb ab 1.28 Urea (mg/dl) T cb bcb abb abb ab abb T ca bcc abc abc ac abc 0.73 T cb bca aba aba aa aba 1.29 Total protein (g/dl) T aa 6.82 ab 7.11 bb 6.83 ab 7.28 cc 7.04 abc T eb 6.62 da 6.73 da 6.29 ca 5.62 ba 5.13 aa 0.17 T ba 6.92 cc 7.31 ec 7.11 db 6.76 bb 6.60 ab 0.15 Albumin (g/dl) T ba 4.18 bc 4.56 dc 3.65 ab 4.34 cc 4.32 cc T db 3.84 ca 3.93 ca 3.30 ba 3.30 bb 2.67 aa 0.14 T ca 4.09 cb 4.25 cb 3.35 ba 2.99 aa 3.06 ab 0.13 Globulin (g/dl) T ab 2.64 aa 2.55 aa 3.18 ca 2.94 bb 2.73 abb T b 2.78 cb 2.81 cb 2.99 da 2.32 aa 2.47 ba 0.12 T a 2.83 bb 3.07 cc 3.76 eb 3.77 ec 3.55 dc 0.17 ALT (IU/L) T T T AST (IU/L) T T T ALP (IU/L) T T T LDH (IU/L) T T T abcde&abc Mean with different superscript with in a row and column, respectively differ significantly (P<0.05) T 1 : Negative control; T 2 : Infected Control; T 3 : Infected Treatment

6 730 INDIAN JOURNAL OF ANIMAL RESEARCH reported in lambs given diets containing CT supplied through Ficus infectoria leaves at 1.5-2% (Dey et al., 2008) and CT supplemented diets through LMM of F. infectoria and P. guajava at 1.5% in H. contortus infected sheep (Pathak et al., 2013a). Similar variations in serum urea level in GIN infected sheep with and without CT supplemented diets from Acacia mearnsii have been reported by Cenci et al. (2007). Total protein, albumin and globulin among three different groups differed significantly (P<0.001, P<0.05). The globulin (g/dl) level was significantly (P<0.004) higher in T 3 group as compared to both T 1 and T 2 groups. The reduction in total protein and albumin in T 2 group and elevated level of globulin in T 3 group might be attributed to the diversion of amino nitrogen and energy from muscle, bone and collagen fibre. This in turn increases the endogenous losses of protein through urinary and faecal excretion. Some of it might be required for the synthesis of specific proteins for tissue repair and for immunological reaction to infection (MacRae, 1993). Present results are in conformity with the findings of many workers (Coop and Holmes, 1996; Van Houtert and Sykes, 1996; Pathak and Tiwari 2013b), who reported that sheep and kids offered a higher plane of nutrition resulting into better withstand of pathological effects of GINs. The activities of ALT, AST, ALP and LDH did not differ significantly (P<0.05) irrespective of groups and were within the normal physiological range (Kaneko, 1997). The CT are not absorbed into the blood stream, therefore, under normal physiological conditions; CT are not likely to damage vital organs such as liver, kidney and spleen. Other studies (Dey et al, 2008; Pathak et al., 2013a) observed similar trends in enzyme activity in lambs and sheep fed tanniferous tree leaves and LMM based diets. FECs: The FECs differed significantly (P<0.05) among groups, periods and interaction between groups and period (Fig. 1). The FECs in T 2 group were increased significantly (P<0.05) throughout the study periods relative to their counterparts in T 3 group. Reduced FECs could be attributed to direct effect of CT on fecundity (reduced), death of adult worms and indirectly by improved immune function against GIN through enhanced in tissue protein supply (Niezen et al., 2002; Shaik et al., 2006;). Alternatively the CT could form a complex with nutrients and inhibit nutrients availability for larval growth or decrease GINs metabolism directly Fig 1: Effect of MNB and MNB-CT supplementation on FECs (per gram) in H. contortus infected goats through inhibition of oxidative phosphorylation (Scalbert, 1991), causing larval death (Athanasiadou et al., 2001). Pathak et al. (2013 b,c) have reported that CT extracts from tree leaves can disrupt the life cycle of GIN by preventing their eggs from hatching and by preventing larval development to the infective stage. Both direct and indirect effects against H. contortus infection appear to be occurring in the present study. The increased level of globulin in T 3 probably reflects an improved rate of immunoglobulin synthesis associated with the development of immunity (Kyriazakis et al., 1994) and an adequate supply of dietary protein or more availability of amino acids in the lower gastrointestinal tract enabled infected animal to improve the capacity to mount an effective immunological response, which enhanced the onset of parasite rejection. CONCLUSION It may be concluded that supplementation of MNB- CT blocks in goats indicates a discernible positive impact on protein bioavailability and decreased H. contortus load. A noticeable encouraging impact was apparent on nutrient intake and utilization and biochemical profile in H. contortus infected goats on MNB-CT supplementation. Incorporation of tanniferous LMM in MNB-CT may be used as alternative feed resource and natural dewormer for the control of H. contortus infection in goats. ACKNOWLEDGEMENT The authors are grateful to Dean, FVSc & AH, SKUAST-J, R. S. Pura, Jammu for providing necessary facilities to carry out the research work. REFERENCES Anderson, N. (1982). Internal parasites of sheep and goats. In: Coop, L.E., ed. Sheep and Goat Production. Elsevier Scientific Publishing Company, pp Anonymous (1986). Manual of Veterinary Parasitological Laboratory Techniques. Bulletin No. 418, Ministry of Agriculture, Fisheries and Food. London. 1986, pp AOAC (1995). Official Methods of Analysis (16 th ed. Vol.I). Association of Official Analytical Chemists. Washington, DC. Athanasiadou, S. Houdijk, J. and Kyriazakis, I. (2008). Exploiting synergisms and interactions in nutritional challenge on appatite, digestibility, rate of passage. Small Rum. Res. 76: 2 11.

7 Volume 50 Issue 5 (2016) 731 Athanasiadou, S. Kyriazakis, I. and Jackson, F. (2003). Can plant secondary metabolites have a role in controlling gastro intestinal nematode parasitism in small ruminants? Proc. VI Int. Symp. Nutr.Herb., October 2003, Merida, Mexico. Athanasiadou, S. Kyriazakis, I. Jackson, F. and Coop, R. L. (2001). Direct anthelmintic effects of condensed tannins towards different gastrointestinal nematodes of sheep: in vitro and in vivo studies. Vet. Parasitol. 99: Aye, P. A. and Adegun, M. K. (2010). Digestibility and growth in West African Dwarf sheep fed gliricidia-based multinutrient block supplements. Agri. Biol. J. N. America. Bergmeyer, H. U. Horder, M. and Rej, R. (1986a). International Federation of Clinical Chemistry (IFCC) Scientific Committee, Analytical Section: Part-2. J. Clin. Chem. Clin. Bioche. 24: Bergmeyer, H.U. Horder, M. and Rej R. (1986b). International Federation of Clinical Chemistry (IFCC) Scientific Committee, Analytical Section: Part-3. J. Clin. Chem. Clin. Bioche. 24: Cenci, F. B. Louvandini, H. McManus, C. M. Dell Porto, A. Costa, D. M. Araujo, S. C. Minho, A. P. and Abdalla, A. L. (2007). Effects of condensed tannin from Acacia mearnsii on sheep infected naturally with gastrointestinal helminthes. Vet. Parasitol. 144: Coop, R. L. and Holmes, P. H. (1996). Nutrition and Parasitic Interaction. Int. J. Parasitol. 26: Coop, R. L. and Kyriazakis, I. (2001). Influence of host nutrition on the development and consequences of nematode parasitism in ruminants. Trends Parasitol. 17: Dey, A. Dutta, N. Sharma, K. and Pattanaik, A. K. (2008). Effect of dietary inclusion of Ficus infectoria leaves as a protectant of proteins on the performance of lambs. Small Rum. Res.75: Doumas, B. T. Arends, R. L. and Pinto, P. C. (1972). Standard Methods of Clinical Chemistry. Academic Press, Chicago. 7: Dschaak, C. M. Williams, C. M. Holt, M. S. Eun, J. S. Young, A.J. and Min, B. R. (2011). Effects of supplementing condensed tannin extract on intake, digestion, ruminal fermentation, and milk production of lactating dairy cows. J. Dairy Sci. 94: Entrocasso, C. M. Parkins, J. J. Armour, J. Bairden, K. and McWilliam, P. N. (1986). Metabolism and growth studies in housed calves given a morantel sustained release bolus and exposed to natural trichostrongyle infection. Res. Vet. Sci. 40: Henry, R. T. Chiamori, N. Goiub, O. J. and Berkman, S. (1960). Revised spectrophotometric methods for the determination of glutamine-oxaloacitic transaminase. Am. J. Clin. Pathol. 34: 381. Kabasa, J. D. Opuda-Asibo, J. and Ter Meulen, U. (2000). The effect of oral administration of polyethylene glycol on faecal helminth egg counts in pregnant goats grazed on browse containing condensed tannins. Trop. Anim. Health Prod. 32: Kaneko, J. J. (1997). Clinical Biochemistry of Domestic Animals. Fifth ed., Academic Press, New York, USA, pp Kyriazakis, I. Olham, J. D. Coop, R. L. and Jackson, F. (1994). The effect of sub clinical intestinal nematode infection on the diet selection of growing sheep. Br. J. Nutr. 72: Lehninger, A. L. Nelson, D. L. and Cox, M. M. (1993). Principles of Biochemistry, 2 nd Edn, CBS Publ. New Delhi. Mohammed, I. D. Baulube, M. and Adeyinka, I. A. (2007). Multinutrient blocks 1: Formulation and production under a semi-arid environment of North East Nigeria. J. Biol. Sci. 7: Niezen, J. H. Charleston, W. A. G. Robertson, H. A. Shelton, D. Waghorn, G. C. and Green, R. (2002). The effect of feeding sulla (Hedysarum coronarium) or lucerne (Medicago sativa) on lambs parasite burdens and immunity to gastrointestinal nematodes. Vet. Parasitol. 105: NRC (2007). Nutrient Requirements of Small Ruminants; Sheep, Goats, Servids and New World Camelids. National Academic Press, Washington DC. Pathak, A. K. (2011). Nutritional status and performance of ruminants as influenced by gastrointestinal nematodes - an overview. N.E. Vet. XIII (3): Pathak, A. K. (2013). Potential of using condensed tannins to control gastrointestinal nematodes and improve small ruminant performance. Int. J. Mol. Vet. Res. 3: MacRae, J. C. (1993). Metabolic consequence of intestinal parasitism. Proc. Nutr. Soc. 52: Makkar, H.P.S. (2000). Quantification of tannins in tree foliage. A laboratory manual for the Joint FAO/IAEA working document, IAEA, Viena. pp 1-26.

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