Variations in plasma biotin and mineral concentrations due to zinc-biotin supplementation in lame Karan Fries cows during peri-parturient period

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1 Indian J. Anim. Res., 49 (6) 2015: Print ISSN: / Online ISSN: AGRICULTURAL RESEARCH COMMUNICATION CENTRE Variations in plasma biotin and mineral concentrations due to zinc-biotin supplementation in lame Karan Fries cows during peri-parturient period Pragya Bhadauria*, S.S Lathwal, Y.S Jadoun 1, Rohit Gupta and Indu Devi ICAR-National Dairy Research Institute, Karnal, Haryana India. Recieved: Accepted: DOI: /ijar.7039 ABSTRACT Nutritional programme of pregnant cows have a significant effect on occurrence of lameness in later lactation. Highly productive lactating cows have much greater needs for minerals and vitamins. Blood biotin and zinc, that are important nutrients in the formation and integrity of the keratinized tissues, often decreases around calving and extra supplementation is recommended, particularly in lame cows during peri-parturient period. The present study was carried out on 40 high yielding lame crossbred KF cows maintained at Livestock Research Centre (LRC), NDRI Karnal, which were distributed into control and three treatment groups and supplemented with 10, 20 and 30 mg/d biotin and 2g of ZnSO 4 during periparturient period. The results of the study suggested that, transition diet with biotin and Zn supplementation significantly increased the concentrations of plasma biotin and Zn levels in supplemental groups compare to control. The overall biotin values were ±11.38, ±24.78, ±29.40 and ±22.77 ng/l and Zn values were 0.40±0.01, 1.12±0.05, 1.22±0.05 and 1.18±0.05 ppm in control, T-1, T-2 and T-3 groups respectively. Thus, by feeding supplemental nutrients (biotin and zinc) lameness can be minimize in high yielding dairy cows during peri-parturient period. Key words: Biotin, Lameness, Peri-parturient period, Supplementation, Zinc. INTRODUCTION Nutritional management including trace mineral supplementation has been identified as a key component in the prevention of lameness, especially during peri-parturient period in high yielding cows. Many physiological changes occur in late gestation and early lactation of the dairy cow that affect nutrient uptake and flow. Macro and trace elements plays an important role in minimizing lameness through their roles in the immune function, production of horn tissue, maintenance of epithelial and connective tissue in dairy animals (Kilic et al., 2007). Trace minerals such as calcium, zinc, copper, cobalt and manganese play an important role in normal hoof health in dairy animals. In vitamins, Biotin is essential for the formation and integrity of the keratinized tissues. In a recent retrospective study, a number of significant predictive relationships were found between serum mineral concentrations in the four weeks prior to or following calving and periparturient diseases (Van Saun et al., 2004; 2005; 2006a,b). It seems that in the last stage of pregnancy and around calving, dams lose more Zn than they ingest, their Zn reserves are depleted and Zn plasma concentrations decrease. (Muehlenbein et al., 2001; Kracmar et al., 2003; Pavlata et al., 2004b). Assessment of Ca concentrations around the time of calving is a useful indicator of how well the Ca regulatory system is working and potential for clinical or subclinical hypocalcemia *Corresponding author s pragyacari@gmail.com. 1 Central Avian Research Institute, Izatnagar. problems (Oetzel, 2004). Deficiencies in concentrations of these nutrients would negatively influence normal maturation of keratinocytes and thus affect the integrity of horn produced during this period (Mulling et al., 1999). The present study was aimed to monitor the concentration of different minerals and biotin as a result of zinc-biotin supplementation for preventing lameness in high yielding cows during periparturient period in KF cows. MATERIALS AND METHODS The study was conducted on lame Karan Fries (KF) cows maintained at Livestock Research Centre (LRC), NDRI Karnal, Haryana during their peri-parturient period (two months before and two months after calving). Animals selected were of different parity (1 st to 5 th ), having different stages of lactation and average milk yield per day. Quasirandom distribution was followed for even distribution of experimental animals. Skilled workers at the Veterinary Dispensary and Animal health Section, NDRI, diagnosed the animals for hoof disorders based on locomotion scoring system given by Sprecher et al., (1997). A total of 40 lame high yielding KF cows were selected and distributed into three treatments and a control group. Supplemented cows were identified with paint marking on their backs. Following diet were offered to the experimental animals up to 60 days before their predicted calving date and 60 days postpartum. Control diet with 2mg of ZnSO 4 and 10 mg of biotin was

2 784 INDIAN JOURNAL OF ANIMAL RESEARCH offered to treatment group-1 (T-1), control diet with 2mg of ZnSO 4 and 20 mg of biotin was offered to treatment group-2 (T-2) and control diet with 2mg of ZnSO 4 and 30 mg of biotin was offered to treatment group-3 (T-3) once a day at the time of morning in the form of Laddoos of jaggery. Locomotion scoring were carried out on a 5-point scale (0 to 5: Normal to severely lame) described by Locomotion Scoring Guide (Sprecher et al.,1997) at 60 days pre-partum and 60 days post-partum. Blood samples were collected on 60d, 15d pre-partum, on the day of calving and 60d, 15d post partum early in the morning (7.00 to 8.00 AM) before feeding. Immediately after collection the blood samples were centrifuged at 3000 rpm for 15 to 20 minutes, plasma were separated and stored in cryovials of 2ml capacity at -20 o C until analysis. Plasma minerals viz. calcium (Ca), Zinc (Zn) and Copper (Cu) were estimated with the help of Atomic absorption Spectrophotometer (Model Z-5000, Polarized Zeeman Atomic absorption Spectrophotometer, Hitachi High-Technologies Corporation, Tokyo, Japan). Biotin was quantitatively determined by enzyme immunoassay (Avidinbinding substances) (Cat #M046019). The data obtained was analysed by using sigmaplot version 11.0, Systat software Inc, USA by using paired t-test. RESULTS AND DISCUSSION Effect of biotin and Zn supplementation on plasma concentrations of biotin (ng/l) during peri-parturient period: The effect of supplementation of biotin and Zn on plasma biotin concentration during peri-parturient period is given in the Table 1. The overall biotin values were ±11.38, ±24.78, ±29.40 and ±22.77 ng/l in control, T-1, T-2 and T-3 group, respectively. Supplementing 10, 20 and 30mg biotin/d and 2gm of Zn, linearly increased plasma biotin concentrations throughout the peri-parturient period as compared to the control. In T-1 group, the cows had significantly lower level of biotin concentration as compared to T-3 on the day of calving. In addition, non-significantly lower biotin concentration was observed in T-1 group compared to T-2 and T-3 group during peri-parturient period, but higher than control group. In control group, there is continuous decline in the plasma biotin concentration due to events associated with parturition and lactation which reduces plasma biotin. The overall biotin concentration was observed significantly (P<0.05) highest in T-3 group, whereas lowest in control group. Mean plasma biotin concentration was increased by supplemental biotin. This increase was dramatic within the first week of supplementation in all the treatments. The result showed that the effect was observed to be significant on day of calving and day 15 th post-partum. Upon calving, biotin concentration increased to a greater degree in cows given supplemental biotin with 30mg/d (T3) compared with T1, T2 but control cows experienced a decline upto 15d pre-partum. The concentration of plasma biotin concentration was found to be highest on 60d postpartum in all the supplemental groups as compare to control. Daily excretions of biotin in milk when cows are lactating could reduce biotin reserves and plasma concentrations. Biotin is involved with metabolic pathways that are important for the production of energy. However, in treatment group s supplementation of biotin at different levels will compensate the excretion of biotin in milk and maintain a higher plasma biotin concentration which indirectly affects the hoof health and promote an early recovery. Plasma biotin concentrations in heifers and nonlactating cows fed 20mg of supplemental biotin/d were 3.6 to 4.8 ng/ml (Steinberg et al.,1995). Plasma biotin baseline levels were ng/l in the controls and ng/ l in the supplemented animals (Frigg et al., 1993). In contrast to this, Zimmerly and Weiss (2001), a greater concentration of plasma biotin was observed at two weeks before parturition than during lactation and highest plasma concentration of biotin at 1d postpartum compared with other time periods. In the present study, for the lactating period, concentrations of biotin in plasma for control cows fed 30mg of supplemental biotin/d were slightly higher than 20mg of supplemental biotin/d as reported by Midla et al., (1998) and Zimmerly and Weiss (2001). Researchers suggested that events associated with parturition altered plasma biotin concentrations in biotin-supplemented cows. Effect of biotin and Zn supplementation on plasma concentrations of minerals during peri-parturient period Plasma zinc concentration: The effect of supplementation of biotin and Zn on plasma Zn concentration during TABLE 1: Effect of biotin and zinc supplementation on plasma biotin concentration (ng/l) during peri-parturient period ±21.60 aa ±17.40 aa ±22.62 aa ±10.35 aa ±25.03 aa ±23.05 ab ±34.60 ab ±8.76 ab ±26.16 aa ±24.95 ab ±38.07 abb ±17.69 bc ±26.61 aa ±25.98 abc ±38.82 abc ±18.07 bd ±30.01 aa ±15.95 bd ±37.22 bd ±24.13 be Overall ±11.38 a ±24.78 b ±29.40 bc ±22.77 c

3 experimental period is presented in the Table 2. The overall Zn values were 0.40±0.01, 1.12±0.05, 1.22±0.05 and 1.18±0.05 ppm in control, T-1, T-2 and T-3 group, respectively. The overall plasma Zn conc. was observed significantly (P<0.05) lower in control group as compared to supplemental groups, while its concentration was highest in T-2 group but not significantly. The results showed that the effect of biotin and Zn supplementation was observed to be non-significant during peri-parturient period between T-1, T-2 and T-3 groups, while differ significantly to control group. The plasma zinc levels remained relatively constant until late in pregnancy when a decline occurred. A more marked decline occurred at calving period in all the supplemental and treatment groups as compared to other periods. These finding were in agreement with the finding of Goff and Stabel (1990) and Xin et al. (1993). It was most likely a consequence of colostrum formation which caused loss of Zn. The late-term fetus accumulates Zn at a rate of about 12 mg/d (House and Bell, 1993); whereas 15 kg of colostrum contains about 285 mg Zn (Kincaid and Cronrath, 1992). This rapid need for Zn in synthesis of colostrum may explain why Zn concentration is lower in blood of cows on day of calving. Increased stress e.g. in association with an acute phase response due to inflammatory reactions in the uterus also reduces plasma Zn concentration. Stress induces synthesis of metallothionein, a protein associated with Zn distribution. As a consequence, Zn is redistributed from blood to other tissues, such as the liver. In addition, glucocorticoids reduce Zn absorption and combine with Volume 49 Issue 6 (December 2015) 785 various stressors to stimulate metallothionein synthesis, which pulls Zn into cells. Plasma calcium concentration: The effect of supplementation of biotin and Zn on plasma Ca concentration during peri-parturient period is presented in the Table 3. The overall Ca values were 10.35±0.09, 10.71±0.09, and 11.14±0.15 and 11.08±0.12 mg/dl in control, T-1, T-2 and T-3 groups respectively. The overall Ca concentration was observed significantly (P<0.05) highest in T-2 group, whereas lowest in control group. The result showed that the effect was observed to be significant on the day of calving and day 15 th post-partum. In control and T-1 group, the cows had significantly lower level of Ca concentration compared to T-2 and T-3 on the day of calving. In addition, nonsignificantly lower Ca concentration was observed in T-3 group compared to T-2 group. Although, total plasma Ca is above the critical limit so no sign of hypocalcaemia was observed in any cows. Physiological fluctuations occur immediately before and after calving in the blood levels of Ca, (Dukes 1993). Blood levels of Ca are expected to decrease at calving due to the large demand of colostrum and milk production. The levels of Ca was significantly (p<0.05) lowered at calving, reaching levels just under the reference values mmol/l (Xin et al., 1993). Plasma Ca concentrations are reduced in early postpartum cows because of increased demand of Ca for synthesis of milk coupled with the relatively slow response in up-regulating Ca absorption from the intestinal tract. The postpartum depression in plasma Ca is greater in older cows than in primiparous cows (Szenci et al., 1994). TABLE 2: Effect of biotin and zinc supplementation on plasma Zn concentration (ppm) during peri-parturient period ±0.03 aa 1.22±0.12 ba 1.42±0.09 ba 1.34±0.14 ba ±0.03 aa 1.32±0.07 ba 1.30±0.09 ba 1.34± 0.08 ba ±0.02 aa 0.76±0.08 bb 0.85±0.05 bb 0.77±0.07 bb ±0.02 aa 0.89±0.11 bb 1.06±0.08 bab 0.94±0.10 bb ±0.03 aa 1.45±0.05 ba 1.51±0.04 ba 1.49±0.06 ba Overall 0.40±0.01 aa 1.12±0.05 b 1.22±0.05 b 1.18±0.05 b TABLE 3: Effect of biotin and zinc supplementation on plasma Ca concentration (mg/dl) during peri-parturient period ±0.08 aa 11.31±0.14 aa 12.23±0.29 ba ±0.23 aba ±0.12 aa 10.91±0.16 aba 11.53±0.27 bab ±0.24 bac ±0.16 abc 10.51±0.14 abab 10.78±0.24 bbc ±0.23 bbc ±0.15 ab 10.08±0.18 abb 10.16±0.27 abc ±0.26 bb ±0.11 aac 10.72±0.13 ab 10.99±0.22 abc ±0.28 ab Overall 10.35±0.09 a 10.71±0.09 b 11.14±0.15 d 11.08±0.12 cd

4 786 INDIAN JOURNAL OF ANIMAL RESEARCH TABLE 4: Effect of biotin and zinc supplementation on plasma Cu concentration (ppm) during peri-parturient period ±0.04 aa 1.06±0.01 aa 1.07±0.01 aa 1.08±0.01 aa ±0.04 aba 0.89±0.04 abb 0.94±0.06 aa 0.47±0.11 bb ±0.03 aac 1.11±0.02 aac 1.13±0.02 ab 1.14±0.01 aac ±0.03 aac 1.15±0.01 aac 1.18±0.03 ab 1.16±0.01 aac ±0.02 abc 1.18±0.01 ac 1.21±0.04 ab 1.19±0.01 ac Overall 1.05±0.02 ac 1.07±0.02 bc 1.11±0.02 b 1.01±0.04 a Plasma copper concentration: The effect of supplementation of biotin and Zn on plasma Cu concentration during peri-parturient period is presented in the Table 4. The overall Cu values were 1.05±0.02, 1.07±0.02, 1.11±0.02, 1.01±0.04 ppm in control, T-1, T-2 and T-3, respectively. The overall Cu concentration was observed significantly (P<0.05) highest in T-2 group whereas lowest in T-3 group. The result showed that the effect of supplementation was observed to be non significant on different groups. The plasma Cu was lowest at 60d prior to calving, higher at calving and 2 months after calving (p<0.001) compared with before calving. The blood Cu status undergoes several changes during the periparturient period. The lower value before calving could be due to the drainage by the fetal liver (Xin et al., 1993). Ward and Spears (1999) suggest that cattle undergoing stressful periods have increased blood levels of Cu and ceruloplasmin, as Cu transport protein ceruloplasmin is considered an acute phase protein and its concentration increases in response to injury, infections and inflammation (Conner et al., 1986). This might be one reason for the increased blood level of this nutrient, as calving is considered a stressful period with tissue damages for example in the uterus. Cu may also improve digital integrity and plays an important role in immune function, in the production of horn tissue, and maintenance of the epithelial and connective tissue, so it may also contribute in early recovery from lameness. REFERENCES Conner, J.G., Eckersall, P.D, Doherty, M and Douglas, T.A. (1986). Acute phase response and mastitis in the cow. Res Vet Sci. 41: Dukes, H.H (1993). Physiology of domestic animals. 11 th edition. Edited by Swenson MJ, Reece WO. Cornell University Press. Ithaca and London Frigg, M., Straub, O.C and Hartmann, D. (1993). The bioavailability of supplemental biotin in cattle. Int. J. Vit. Nutr. Res. 63: Goff, J.P and Stabel, J.R. (1990). Decreased plasma retinol, alpha-tocopherol, and zinc concentration during the periparturient period: effect of milk fever. J. Dairy Sci. 73: House, W.A and Bell, A.W. (1993). Mineral accretion in the fetus and adnexa during late gestation in Holstein cows. J. Dairy Sci. 76: Kilic, Nuh., Ceylan. Ahmet., Serin. Ilker and Gokbulut, Cengiz. (2007). Possible interaction between lameness, fertility, some minerals, and vitamin E in dairy cows. Bull Vet Inst Pulawy 51: Kincaid R.L, Cronrath J.D. (1992). Zinc concentration and distribution in mammary secretions of peripartum cows. J. Dairy Sci. 75: Kracmar, S., Gajdusek, S., Jelinek, P and Illek J. (2003). Changes in contents of some macro- and microelements in goat s colostrum within the first 72 h after parturition. Small Ruminant Research. 49: Midla, L.T., Hoblet, K.H., Weiss, W.P and Moeschberger, M.L. (1998). Supplemental dietary biotin for prevention of lesions associated with aseptic subclinical laminitis (pododermatitis aseptica diffusa) in primiparous cows. Am J Vet Res. 59: Muehlenbein, E.L., Brink, D.R., Deutscher, G.H., Carlson, M.P and Johnson A.B. (2001). Effects of inorganic and organic copper supplemented to first-calf cows on cow reproduction and calf health and performance. Journal of Animal Science. 79: Mulling, C., Bragulla, H., Reese, S., Budras, K. D and Steinberg, W. (1999). How structures in bovine hoof epidermis are influenced by nutritional factors. Anat. Hist. Embryol. 28: Oetzel, G.R. (2004). Monitoring and testing dairy herds for metabolic disease. Vet Clinics NA: Food Anim Prac. 20:

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