Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

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Effect of Dietary Supplementation of Seaweed Formulations on Nutrient Utilization, Growth Performance, Serum Antioxidant Activity and Immunity in Crossbred Calves

Bornalee Handique1,*, P. Singh1, A.K. Verma1
1Division of Animal Nutrition, ICAR-Indian Veterinary Research Institute Izatnagar, Bareilly-243 001, Uttar Pradesh, India.
Background: Seaweeds are rich in complex carbohydrates, proteins and low molecular weight nitrogenous compounds, lipids, organic minerals, vitamins and pigments. The aim of the study was to evaluate the effect of seaweed formulations on nutrient utilization, growth performance, serum antioxidant activity and cell mediated immune response of crossbred calves. 

Methods: Eighteen male crossbred calves (130±7.56 kg body weight) were divided into 3 groups of 6 each based on complete randomized design. Concentrate mixture was formulated with crushed maize, de-oiled soybean meal, wheat bran, mineral mixture and common salt for feeding of calves in control (T0) group, while treatment groups T1 and T2 were supplemented with AF-KWP or AFRD-5 at the rate of 4% of concentrate mixture with 1% calcium carbonate and 1% dicalcium phosphate replacing mineral mixture and common salt offered in T0. Experiment was conducted for a period of 240 days. Blood was collected from each calve at 0, 100 and 200 days of experiment. At the end of the experimental feeding a metabolism trial was conducted and cell mediated immune response was asassed. 

Result: There was no significant difference between groups regarding nutrient utilization, total dry matter intake, gain in body weight and feed conversion ratio. However, significant (P<0.001) differences were observed in serum antioxidants status. Moreover, the mean values for skin thickness differed significantly (P<0.001) among different groups and values were 14.03, 15.54 and 16.62 in T0, T1 and T2 group, respectively. Hence seaweed formulations might improve antioxidant status and cell mediated immune response without affecting the nutrient utilization and growth performance of crossbred calves.
Seaweeds are rich in proteins, lipids, polysaccharides, mineral, vitamins and enzyme (Rimber 2007). In contrast to conventional mineral supplements, seaweed is unique in being of plant origin containing a wide range of naturally balanced chelated minerals, trace elements, amino acids and vitamins. Being totally natural and of vegetable origin seaweed is easily digested and is safely fed to animals of all ages (Sykes 2009). There are 3 different groups of seaweed on the basis of thallus colour brown, red and green seaweed. They are also different in many ultra structural and bio-chemical features including photosynthetic pigments, storage compounds, composition of cell walls and presence or absence of flagella.

Seaweeds are rich source of Na, K, Mg, Cl, S, P, I, Fe, Zn, Cu, Se and Mo (Okab et al., 2013). The use of seaweeds in livestock feed increased growth rate and feed conversion efficiency in ruminants (Chowdhury et al., 1995). Many species of macro-algae have anti-bacterial, anti-viral, anti-oxidant and anti-inflammatory properties that improve animal health and function (Bach et al., 2008). Many researchers reported that phlorotannins which is derived from the marine brown algae have strong antioxidant properties (Heo et al., 2005; Shibata et al., 2008). Allen et al., (2001) reported that supplementation of seaweed meal enhanced immunity in beef cattle.

Limited information is available about feeding and evaluation of seaweed in crossbred calves. Therefore, present experiment was designed to assess the effect of seaweed formulations on nutrient utilization, growth performance, serum antioxidant activity and immunity of crossbred calves.
Eighteen male crossbred (Holstein Friesian ´ Indigenous) calves (130±7.56 kg live weight) were divided into 3 groups of 6 each based on CRD. The concentrate mixture was formulated with crushed maize, de-oiled soybean meal, wheat bran, mineral mixture and common salt for feeding of calves in control (T0) group. While the concentrate mixture of calves in groups T1 and T2 were supplemented with AF-KWP and AFRD-5 at 4% with 1% calcium carbonate and 1% dicalcium phosphate, respectively replacing mineral mixture and common salt (Table 1). AF-KWP contains Kapaphycus alvarezi (thrashed weed), Gracilaria salicornia (washed weed) and Kapaphycus water extract at a ratio of 1:1:1 while AFRD-5 comprise of Kapaphycus alvarezi (thrashed weed), Gracilaria salicornia (washed weed) and Turbinaria conoides at a ratio of 2: 2: 1. The experimental calves were offered feed to meet out nutrient requirements as per Icar (2013) feeding standard recommendation for a body weight gain of 500 g/day. All the animals were provided fresh and clean drinking water twice ad lib. The feeding trial was conducted for 240 days. Roughage to concentrate ratio was 50:50. Available green fodder was given to experimental calves to meet out the Vitamin A or carotene requirement.

Table 1: Ingredient compositions of concentrate mixture.



A record of feed intake and live weight was maintained for assessing the growth performance of calves during the entire experimental periods. A metabolism trial of 9 days was conducted at the end of the experimental feeding including 3 days of adaptation and 6 days of collection. Faeces and urine voided in 24 hour by each animal were collected and processed for storage and analysis. A separate aliquot of faecal samples was preserved with dilute (25% v/v) sulphuric acid for nitrogen (N) estimation. Similarly, urine sample was collected under acidic (dilute sulphuric acid) condition and sampled for estimation of N. Feeds, faeces and seaweed samples were also analyzed for the proximate principles (Aoac 2012) and fibre fractions viz. NDF and ADF (Van Soest et al., 1991). 

Blood was collected on 0, 100 and 200 days of the experimental period from each calf by puncturing jugular vein in the morning (before watering and feeding). Blood samples were brought to laboratory and centrifuged at 3000 rpm for 15 minutes to separate serum and stored at -20°C until further analysis. SOD and Catalase concentration was determined by using Cayman’s Superoxide Dismutase Biont Assay ELISA kits based on the biotin double antibody sandwich technology. Glutathione peroxidase was determined by cellular activity assay

The cell mediated immune response was estimated in crossbred calves through in vivo delayed type hypersensitivity (DTH) reaction against phytohaemagglutinin- P (PHA-P). All the calves were injected intra-dermally with 125 μl of either saline solution (8.5 g NaCl) as a negative control on one side and PHA-P (20 μg 125 μ1-1 PHA-P solution) on the other side of neck region. The thickness of the skin was subsequently measured at 0, 24, 48, 72 and 96 hrs.

The statistical analyses were performed by using SPSS computer package (SPSS version 20.0, SPSS Inc., Chicago, USA). The data were statistically analyzed by using ANOVA with Duncan’s post hoc testing to compare experimental groups. For all statistical analyses, probability values less than 0.05 were considered as significant.
Seaweed formulations (AF-KWP and AFRD-5) were higher in total ash and insoluble ash content (Table 2). Total ash, which mainly represents mineral content, in the present experiment was higher than the values reported by Cabrita et al. (2016) with Ulva sp. and G. vermiculophylla. EE content was in agreement with many studies (Khairy and El-Shafay 2013; Mouritsen et al., 2013). The CP content of concentrate mixture was almost similar. The chemical composition of maize fodder and wheat straw was within the normal range reported by Ranjhan, 1988. Our results are also similar to Abdoun et al., (2013) who reported that NDF and ADF were less in seaweed (Ulva lactuca) supplemented group than control.

Table 2: Chemical composition (%) of seaweed formulations, concentrate mixture and roughage.



The intake and digestibility of DM, OM, CP, EE, total carbohydrate, ADF and NDF was comparable in all three groups (Table 3). Therefore it is evident that supplementation of seaweed formulations has no adverse impact on overall rumen microbial and digestive efficiency of the animals. This corroborates well with the findings of Berry and Turk (1944) who have reported no influence of brown seaweed supplementation on DMI. Our findings were in contrary to Antaya et al., (2016) who have reported increased digestibility of OM and NDF due to supplementation of Ascophyllum nodosum.

Table 3: Intake and digestibility of nutrients in different groups.



The results showed that all the experimental calves were in positive Ca, P and N balance (Table 4). The Ca intake (g/d) was significatly (P<0.001) higher in the seaweed supplemental group than control. However, P intake (g/d) was significatly lower (P<0.001) in the seaweed supplemental group than control. The N intake was similar in all 3 groups. The retention (% intake) of Ca and P was significant (P<0.05) among the different groups. Munde (2018) reported that intake (g/d), retention (% intake) of Ca and P were similar among the groups by feeding Kappaphycus and Gracilaria. Munde (2018) also reported that retention of N (% intake) did not differ significantly among the group by feeding Kappaphycus and Gracilaria. Similarly, Venketeswaran (2018) reported that daily intake and retention of N were statistically similar in 3 groups by feeding 2% and 4% Turbinaria conoides.

Table 4: Balances of calcium, phosphorus and nitrogen in different groups.



Non-significant effect was observed in total gain in body weight, ADG, total DMI and FCR among the 3 treatment groups (Table 5). Our results were in agreement with Anderson et al., (2006) who did not observed any influence of Ascophyllum nodosum supplementation on growth performance of crossbred cattle. Our results are in contarary with many studies who observed that seaweed supplementation improved body weight gain (Al-Shorepy et al., 2001; Turner et al., 2002).

Table 5: Growth performance of crossbred calves in different groups.



The level of different antioxidant parameters were presented in Table 6. In the present study, seaweed formulations supplementation increased the activity of SOD, catalase, MDA and GPx. Our findings were similar to Allen et al., (2001) and Saker et al., (2004) who reported that inclusion of Ascophyllum nodosum improved antioxidant status in cattle. The seaweed formulation AFRD-5 used in the present experiment contained a brown seaweed thrashed powder (Turbinaria conoides) which contain phlorotannin, has antioxidant activities. Moreove extracts of Kappaphycus sp. showed higher reducing power and- OH radical scavenging activity than other antioxidants (Holdt and Kraan 2011). Sulfated polysaccharide from Gracilaria sp. exhibited moderate effect in inhibits the formation of free radicals. Kapaphycus alvarezii contains β-carotene, vitamin C and vitamin E which have antioxidant activity (Mantajun et al., 2010). Our result is in contradiction with Abdoun et al., (2013) who reported that seaweed inclusion did not alter the blood antioxidant activity. Similarly, Venketeswaran (2018) reported non-significant effect in serum SOD and GPx concentrations by supplemementation of Turbinaria conoides in crossbred calves.

Table 6: Serum antioxidant status of crossbred calves in different groups.



The mean values for skin thickness differed significantly (P<0.001) among different groups and significantly (P<0.001) maximum skin thickness was observed in group T2 followed by T1 and T0 groups (Fig 1). Maximum average thickness was observed after 24 h and then gradually reduced. Our results are in agreement with Allen et al., (2001) who have observed that Ascophyllum nodosum improved immune response in mammals. Kuznetsova et al., (2015) also observed that fucoidans are agonists for receptors of innate immunity and potent inducers of the cell-mediated immune response. In the present experiment, increased GPx and SOD activity was observed which might contributed to increased cellular immunity. This is similar with the finding of Fike et al., (2001) who have reported increased SOD in ruminants with supplementation of brown seaweed. Kappaphycus and Gracilaria are rich in Se and vitamin E which were positively linked to cellular immunity (Fike et al. 2001). Our results are in agreement with Venketeswaran (2018) and Munde (2018) who reported that cell mediated immune response was significantly higher seaweed supplemented groups.

Fig 1: Absolute skin thickness (mm) after injection of PHA-p at different hours in different groups.

Present study concluded that calcium and phoshporus added seaweed formulations improves antioxidant status and cell mediated immune response without affecting growth performance and nutrient utilization in crossbred calves.
None

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