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Effect of Rumen Protected Methionine and Choline on Blood Biochemical Metabolites, Milk Yield and its Composition during Transition Period in Surti Buffaloes
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First Online 14-10-2021|
Methods: Twenty-seven pregnant multiparous Surti buffaloes in three groups (n=9) from -15 d to 30 d postpartum received supplementation as: T1 (Control: basal diet), T2 (basal diet + RPM@10 gm/animal/day) and T3 (basal diet + RPM@10 gm/animal/day + RPC@ 50 gm/animal/day). Sample was collected at beginning, 1st, 3rd and 6th week for blood and at 1st, 3rd, 6th, 9th and 12th week postpartum for milk. Milk yield was recorded upto 100 days postpartum.
Result: Postpartum TC, HDL and VLDL differed significantly (P<0.05) being highest in T3 and lowest in control (T1) whereas it was reverse for NEFA and BHBA. Supplemented groups had significantly lower TG levels at 1st and 3rd week postpartum. Milk fat upto 9th and SNF, protein, lactose, TAS, Ca, P and Mg upto 6th week were significantly (P<0.05) highest in T3, followed by T2 and T1. It was concluded that RPC along with RPM supplementation is more beneficial than only RPM supplementation in terms of enhancing liver health, reducing negative energy balance and improving milk quality.
Methionine and choline supplementation play metabolic roles especially during transition. They are lipotropic, act as methyl group donor, enhance fat metabolism and minimizes risk of fatty liver and ketosis. Their limited dietary availability due to extensive microbial degradation in rumen (Zhou et al., 2016 and Potts et al., 2020) can be overcome by resistant coating. Therefore supplementing RPM and RPC may meet daily demand of methyl group, improve milk quality and health of dairy animals during transition. Considering the benefits of dietary supplementation of rumen protected forms of methionine and choline (RPM and RPC) and the dearth of studies on these amino acids in buffaloes present study was planned to evaluate blood biochemical, milk yield and its composition on supplementation of rumen protected methionine (RPM) and choline (RPC) in transition Surti buffaloes.
MATERIALS AND METHODS
10 ml blood was collected in K3EDTA and vacutainers for serum at 15 days prepartum and 1st, 3rd and 6th week postpartum. Vacutainers were centrifuged (3000 rpm for 15 minutes) to separate plasma and serum and stored at -20°C until further analysis. Plasma was used to estimate non-esterified fatty acid (NEFA) (Shipe et al., 1980) and serum was used to estimate total cholesterol (TC), high density lipoprotein (HDL), low density lipoprotein (LDL), very low density lipoprotein (VLDL), triglyceride (TG) and β-hydroxy butyric acid (BHBA) by Randox kit.
Daily milk yield was recorded upto 100 days postpartum. 100 ml milk was collected at 1st, 3rd, 6th, 9th and 12th week postpartum for analysing milk composition, total antioxidant status (TAS), macro and micro elements. Milk fat, solid not fat (SNF), lactose, protein, total solids (TS), urea, free fatty acids (FFA) and lactic acid (LA) were estimated using fully automated FOSS made milkoscan FT1 milk analyser (FOSS, Slangerupgade 69, Denmark) at Surat Milk Union Limited (SUMUL), Surat.
Milk serum (whey) was separated (Naser et al., 2014) for estimation of milk macro elements such as calcium (Ca), phosphorus (P), magnesium (Mg), sodium (Na) and potassium (K). Milk Ca, P, Mg were estimated using Randox kits and Milk Na and K were estimated by flame photometer 128 (SYSTRONICS). Milk micro elements were determined by ashing milk samples in muffle furnace, solubilizing with diluted HCL (1:4) and making final volume 100 ml with triple glass distilled water. Milk iron (Fe), manganese (Mn), copper (Cu) and zinc (Zn) were estimated by atomic absorption spectrophotometer (Model AAS141, Electronics Corporation of India Ltd.). Milk TAS was estimated by method of Alyaqoubi et al., (2014). Extraction of antioxidant compounds in fresh milk was carried out according to method of Li et al., (2007).
Data was analysed statistically by ANOVA using DMRT. Means were compared at P<0.05 (Snedecor and Cochran, 1994).
RESULTS AND DISCUSSION
Blood biochemical parameters
Among blood biochemical parameters (Table 2), postpartum TC, HDL and VLDL differed significantly (P<0.05) being highest in T3 and lowest in control (T1) whereas it was reverse for NEFA and BHBA. Supplemented groups had significantly lower TG levels at 1st and 3rd week postpartum. LDL levels differed non-significantly between groups.
TC, HDL, LDL, VLDL and TG are lipid profile parameters. Cholesterol an important inflammatory biomarker (Osorio et al., 2014; Batisel et al., 2017) acts as precursor of steroid hormone that aids revival of postpartum ovarian activity. Increase in HDL might occur due to synthesis of paroxonase in liver that protects HDL from oxidative damage (Turk et al., 2004). Lower TG and higher VLDL in treatment groups might be due to RPM and RPC acting as lipotropic agent thus preventing TG accumulation in liver and stimulating hepatic VLDL formation (Sun et al., 2016).
Plasma NEFA and BHBA are indicators of energy status in dairy animals especially during transition. Adipose tissue releases NEFA into circulation that is transported to liver for metabolism and energy production. TG can either be accumulated in hepatocyte or can be exported out of liver in the form of VLDL (Sun et al., 2016). Hepatic accumulation of TG and increased BHBA in plasma causes fatty liver condition during transition period, which further reduces lactation performance, impairs reproduction and causes various metabolic disorders (Tsiplakou et al., 2017). RPM+RPC causes decrease in NEFA that may be due to better metabolism and its use in energy production. Further, it may also prevent increase in triglyceride in liver and protects liver health during transition.
Milk yield and composition
Among milk parameters (Table 3, 4 and 5), RPM and RPC supplementation had non-significant effect on milk yield. However, Milk fat upto 9th and SNF, protein, lactose, TAS, Ca, P and Mg upto 6th week were significantly (P<0.05) highest in T3, followed by T2 and T1. Remaining parameters did not differ significantly between groups.
MF percentage is reported to increase as acetate to propionate ratio (A:P) ratio increases up to 2.2 (Davis, 1967). Methionine and choline supplementation increases the A:P ratio and enhances MF (Ray et al., 1983). Observed higher MF percentage in T3 and T2 groups might be due to increased ruminal A:P ratio and/or it might have helped mammary fat synthesis. Similar findings for MF were observed by Amrutkar et al., (2015), Potts et al., (2020) and Mavrommatis et al., (2021).
Methionine is limiting amino acid (AA) for milk protein (MP) synthesis (NRC, 2001). Choline supplementation spares methionine for MP synthesis. RPM and RPC supplementation enhances MP content in transition dairy cows and improves efficiency of MP synthesis (Sun et al., 2016). Higher MP percentage in treatment groups might be due to supplemental effect of RPM and RPC in transition Surti buffaloes. Similar results were observed by Osorio et al., (2014), Amrutkar et al., (2015), Sun et al., (2016), Zhou et al., (2016) and Mavrommatis et al., (2021). RPM has been shown to have direct stimulatory effect on lactose biosynthesis in mammary gland (Amrutkar et al., 2015). Higher milk lactose percentage in present study may be attributed to higher blood glucose level in T3 and T2 group that helps in mammary lactose biosynthesis. Milk SNF primarily consists of lactose, caseins, whey protein and minerals. Increased milk SNF at week 2 was consistent with the increased milk yield of protein and lactose (Socha et al., 2005). Observed higher (P<0.05) milk SNF percentage in treatment groups might be due to higher milk protein and lactose percentage. Milk Ca, P and Mg level in T3 and T2 groups might be due to their optimal availability in blood. Methionine and choline plays an important role in maintaining antioxidant level during transition period in dairy animals (Tsiplakou et al., 2017). Hence in present study higher milk TAS levels in treatment groups indicate that methionine and choline elevates TAS of milk that may help in improving keeping quality of milk.
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