TY - JOUR
AU - Lindemann, M. D.
AB - ABSTRACT Piglets are born with purportedly low plasma vitamin D levels. The objective of this study was to investigate the effect of fat-soluble vitamin administration, primarily vitamin D, by different administration routes on plasma vitamin concentrations in suckling pigs. A total of 45 pigs from 5 litters were allotted at birth to 3 treatments within each litter. Pigs were administered 400 IU of α-tocopherol, 40,000 IU of retinyl palmitate, and 40,000 IU of vitamin D3 at d 1 of age either orally or by i.m. injection and compared with control pigs with no supplemental vitamin administration. Blood samples were collected at d 0 (initial), 1, 2, 3, 4, 6, 9, 14, and 20 after administration. Plasma 25-hydroxycholecalciferol (25OHD3), α-tocopherol, retinyl palmitate, and retinol concentrations were analyzed. Except for retinol, the effects of treatment, day, and day × treatment interaction (P < 0.01) were observed on plasma vitamin concentrations. Plasma concentrations of 25OHD3 and α-tocopherol increased immediately regardless of administration routes to peak at d 2 and 1 after administration, respectively. Plasma retinyl palmitate concentrations increased only with the injection treatment, with the peak at d 1 after administration. Plasma concentrations of 25OHD3 in both administration treatments and α-tocopherol in the injection treatment were maintained at greater levels than those in the control treatment until d 20 after administration. With regard to the pharmacokinetic parameters for plasma 25OHD3 concentrations, the injection treatment had greater elimination half-life (P < 0.01), maximum plasma concentrations (P < 0.05), and all area under the curve parameters (P < 0.01) but a lower elimination rate constant (P < 0.01) than the oral treatment. Relative bioavailability of oral administration compared with injection administration was 55.26%. These results indicate that plasma status of 25OHD3,α-tocopherol, and retinyl palmitate are differentially changed between types of vitamins administered and between administration routes and that the injection route had a greater increase and slower disappearance of plasma vitamin levels than the oral route during the suckling period. INTRODUCTION Fat-soluble vitamins are important micronutrients in pigs for normal physiological functions related with vision, immunity, bone integrity, and antioxidant properties (McDowell, 2000). Piglets are born with purportedly low levels of vitamin A, D, and E in plasma due to insufficient placental transfer (Horst and Littledike, 1982; Håkansson et al., 2001). Even though colostrum contributes to slight increases in plasma vitamin concentrations of newborn pigs, plasma levels are reported to either decline or be relatively constant through weaning (Håkansson et al., 2001; Madson et al., 2012). Our previous research (Jang et al., 2014) demonstrated that vitamin D3 administration either orally or by i.m. injection (in a complex with variable vitamin A and E concentrations) to newborn pigs increased plasma 25-hydroxycholecalciferol (25OHD3) concentrations at d 10 after administration regardless of administration routes and that pigs in the i.m. injection route had greater plasma 25OHD3 concentrations than those in the oral route. Additionally, it has been reported that only 50% of an oral dose of vitamin D is absorbed (McDowell, 2000). Several previous studies in pigs have reported increased plasma levels of retinol (Ching et al., 2002), 25OHD3 (Flohr et al., 2014), and α-tocopherol (Hill et al., 1999) by vitamin administration/supplementation, respectively. However, there is no information available that investigates whether there are different responses in plasma vitamin status between types of fat-soluble vitamins administered and between administration routes on temporal plasma vitamin profiles. In this study, we hypothesized, primarily for vitamin D3 administration, that 1) administration routes between oral and injection have different responses and efficiencies to enhance plasma 25OHD3 concentrations and 2) plasma vitamin D status is changed after fat-soluble vitamin administration differentially from plasma vitamin A and E status. Therefore, the objective of this study was to investigate the effects of vitamin D3 administration on temporal plasma vitamin status in suckling pigs combined with vitamin A and E administration. MATERIALS AND METHODS Experiments were conducted at the University of Kentucky, Lexington, under protocols approved by the University of Kentucky Institutional Animal Care and Use Committee. Animals and Experimental Design A total of 45 crossbred (Yorkshire × Landrace × Duroc) pigs were used from 5 litters. At birth, 9 pigs within each litter were assigned to 3 treatments based on gender and BW in a randomized complete block design. Treatments were 1) no supplemental vitamins (control), 2) i.m. injection of 0.8 mL of a vitamin complex (VITAL E-NEWBORN, containing 500 IU of vitamin E as D-α-tocopherol, 50,000 IU of vitamin A as retinyl palmitate, and 50,000 IU of vitamin D3 per mL), and 3) oral administration of 0.8 mL of a vitamin complex (EMCELLE NEWBORN EAD, containing 500 IU of vitamin E as D-α-tocopherol, 50,000 IU of vitamin A as retinyl palmitate, and 50,000 IU of vitamin D3 per mL). Vitamin administration was conducted at d 1 of age. The commercial products used were from Stuart Products Inc. (Bedford, TX). Housing, Diets, and Vitamin Administration Housing for sows and piglets, processing of piglets, and the method for vitamin administration followed were as described by Jang et al. (2014). All sows with pigs were kept in individual farrowing crates in an environmentally controlled farrowing facility without windows. A lactation diet (Table 1) that contained 1,320 IU of vitamin D3 was provided to all sows ad libitum and water was freely available from a water nipple throughout the entire experimental period. All pigs were processed at birth (within 15 h) and assigned to a treatment. Processing of the piglets involved weighing, ear notching, needle teeth clipping, and iron injection with 100 mg as iron dextran. With regard to administration of treatments, pigs were administered vitamin products by either i.m. injection or oral placement in the back of the mouth. The injectable product was provided to each pig in the trapezius muscle on the opposite side of the neck from where the iron injection was given. The orally administered treatment was provided through a plastic tube attached to a 3-mL syringe into which the proper dosage had been drawn. Body weight of the pigs was recorded at d 0, 11, and 20 (average d 19.6) after administration to calculate growth performance. Table 1. Formulation and chemical composition of the lactation diet for sows (as-fed basis) Item  Lactation diet  Ingredient, %      Corn, ground  67.56      Dehulled soybean meal, 48% CP  25.60      Alfalfa meal  2.50      Choice white grease  1.00      Dicalcium phosphate  1.21      Limestone, ground  0.89      Salt  0.50      Vitamin1 and trace mineral2 premix  0.15      Choline chloride, 50%  0.10      Dynamate3  0.50      Total  100.00  Calculated composition      ME, kcal/kg  3,290      CP, %  18.19      Total Lys, %  0.97      Total Ca, %  0.75      Total P, %  0.60  Item  Lactation diet  Ingredient, %      Corn, ground  67.56      Dehulled soybean meal, 48% CP  25.60      Alfalfa meal  2.50      Choice white grease  1.00      Dicalcium phosphate  1.21      Limestone, ground  0.89      Salt  0.50      Vitamin1 and trace mineral2 premix  0.15      Choline chloride, 50%  0.10      Dynamate3  0.50      Total  100.00  Calculated composition      ME, kcal/kg  3,290      CP, %  18.19      Total Lys, %  0.97      Total Ca, %  0.75      Total P, %  0.60  1Supplied per kilogram of diet: 6,600 IU vitamin A, 1,320 IU vitamin D3, 66 IU vitamin E, 6.6 mg vitamin K (menadione sodium bisulfate complex), 8.8 mg riboflavin, 22 mg D-pantothenic acid, 88 mg niacin, 6.6 mg vitamin B6, 33 μg vitamin B12, 220 μg d-biotin, and 1,320 μg folic acid. 2Supplied per kilogram of diet: 100 mg Zn as ZnO, 120 mg Fe as FeSO4·H2O, 45 mg Mn as MnO, 12 mg Cu as CuSO4·5H2O, 1.5 mg I as CaI2O6, and 0.30 mg Se as NaSeO3. 3The product contained 180 g of K, 110 g of Mg, and 220 g of S per kilogram (Mosaic Feed Ingredients, South Riverview, FL). View Large Table 1. Formulation and chemical composition of the lactation diet for sows (as-fed basis) Item  Lactation diet  Ingredient, %      Corn, ground  67.56      Dehulled soybean meal, 48% CP  25.60      Alfalfa meal  2.50      Choice white grease  1.00      Dicalcium phosphate  1.21      Limestone, ground  0.89      Salt  0.50      Vitamin1 and trace mineral2 premix  0.15      Choline chloride, 50%  0.10      Dynamate3  0.50      Total  100.00  Calculated composition      ME, kcal/kg  3,290      CP, %  18.19      Total Lys, %  0.97      Total Ca, %  0.75      Total P, %  0.60  Item  Lactation diet  Ingredient, %      Corn, ground  67.56      Dehulled soybean meal, 48% CP  25.60      Alfalfa meal  2.50      Choice white grease  1.00      Dicalcium phosphate  1.21      Limestone, ground  0.89      Salt  0.50      Vitamin1 and trace mineral2 premix  0.15      Choline chloride, 50%  0.10      Dynamate3  0.50      Total  100.00  Calculated composition      ME, kcal/kg  3,290      CP, %  18.19      Total Lys, %  0.97      Total Ca, %  0.75      Total P, %  0.60  1Supplied per kilogram of diet: 6,600 IU vitamin A, 1,320 IU vitamin D3, 66 IU vitamin E, 6.6 mg vitamin K (menadione sodium bisulfate complex), 8.8 mg riboflavin, 22 mg D-pantothenic acid, 88 mg niacin, 6.6 mg vitamin B6, 33 μg vitamin B12, 220 μg d-biotin, and 1,320 μg folic acid. 2Supplied per kilogram of diet: 100 mg Zn as ZnO, 120 mg Fe as FeSO4·H2O, 45 mg Mn as MnO, 12 mg Cu as CuSO4·5H2O, 1.5 mg I as CaI2O6, and 0.30 mg Se as NaSeO3. 3The product contained 180 g of K, 110 g of Mg, and 220 g of S per kilogram (Mosaic Feed Ingredients, South Riverview, FL). View Large Blood Sampling and Chemical Analysis Blood samples, relative to the treatment administration, were collected from the jugular vein of piglets at d 0 (before administration of any treatments), 1, 2, 3, 4, 6, 9, 14, and 20 after administration. Blood samples were centrifuged at 1,700 × g at 4°C for 20 min; plasma samples were then aliquoted into microtubes and stored at –20°C until analysis. Before analysis, plasma samples were pooled by treatment within a litter. Plasma samples were then sent to the Iowa State University Veterinary Diagnostic Laboratory (Ames, IA) and then analyzed for α-tocopherol, retinyl palmitate, and retinol concentrations; samples were subsequently sent to Heartland Assays (Ames, IA) to be analyzed for 25OHD3 concentrations. Detection limits for 25OHD3, α-tocopherol, and retinyl palmitate were 2.5 ng/mL, 0.5 μg/mL, and 0.01 μg/mL, respectively. Pharmacokinetics for Plasma 25-Hydroxycholecalciferol Concentrations Pharmacokinetic parameters for the oral and injection routes were obtained with GraphPad Prism version 5.00 for Windows (GraphPad Software, San Diego, CA) and Microsoft Office Excel (Microsoft, Redmond, WA). Time-dependent plasma concentration profiles of 25OHD3 were analyzed for each litter with a 1-compartment open model with first-order absorption and elimination to determine the absorption rate constant (ka; d–1) and the elimination rate constant (kel; d–1). The half-life of absorption (ta1/2; d) and the half-life of elimination (tel1/2; d) were determined by 0.693/ka and 0.693/kel, respectively. The time to reach maximum concentration (Tmax; d) was determined as [ln(ka) – ln(kel)]/(ka – kel), and the maximum concentration (Cmax; ng/mL) was determined by the concentration at Tmax. Area under the curve (AUC) from d 0 to the last sampling day (AUC0–20; ng/mL∙d) was calculated using the trapezoidal rule and the extrapolated AUC from the last sampling day to infinity (AUC20–∞; ng/mL∙d) was determined as the concentration at the last sampling day divided by kel. The total AUC from d 0 to infinity (AUC0–∞; ng/mL∙d) was calculated by summing AUC0–20 and AUC20–∞. Relative bioavailability between the oral and injection routes of administration was determined as total AUC0–∞ for oral/total AUC0–∞ for injection. Statistical Analysis Growth data were analyzed by ANOVA for a randomized complete block design using PROC MIXED of SAS (version 9.2; SAS Inst. Inc., Cary, NC). Models included the treatment as a fixed effect and the litter as a random effect. All plasma data was subjected to repeated measures ANOVA to detect the effect of treatment, day, and day × treatment interaction using PROC MIXED of SAS with a heterogeneous autoregressive covariance structure. The experimental unit was the litter. Assay values below the limit of detection were treated as missing values. Statistical outliers within each treatment and day were identified using the Grubb's test outlier calculator (GraphPad Software) and excluded from the data analysis. Least squares means were separated using the PDIFF option of SAS when there was an interaction between day and treatment. The pharmacokinetic parameters of plasma 25OHD3 concentrations obtained from each individual litter for oral and injection routes were analyzed by ANOVA for a randomized complete block design using PROC MIXED of SAS and separated using the PDIFF option of SAS. Models included the treatment as a fixed effect and the litter as a random effect. Statistical differences were considered significant at P < 0.05 and tendency at P < 0.10. RESULTS There was no difference on growth performance among administration treatments (Table 2). For plasma 25OHD3 concentrations, a treatment effect was observed along with effects of day and day × treatment interaction (P < 0.01; Table 3) in which plasma 25OHD3 concentrations in the vitamin-administered pigs increased immediately, peaked at d 2 after administration, and then decreased continuously until d 20 after administration regardless of administration routes. The injection route had the greatest values compared with the control and oral treatments, and the oral treatment had greater values than the control treatment for all days monitored (P < 0.05). Table 2. Effect of administration routes of fat-soluble vitamins on growth performance for suckling pigs1   Treatment3    Day2  Control  Oral  Injection  SEM  BW, kg      d 0  1.87  1.85  1.88  0.07      d 11  4.52  4.50  4.49  0.24      d 204  7.03  7.09  7.06  0.46  ADG, kg/d      d 0 to 11  0.24  0.24  0.24  0.02      d 11 to 20  0.29  0.30  0.30  0.03      d 0 to 20  0.26  0.27  0.26  0.02    Treatment3    Day2  Control  Oral  Injection  SEM  BW, kg      d 0  1.87  1.85  1.88  0.07      d 11  4.52  4.50  4.49  0.24      d 204  7.03  7.09  7.06  0.46  ADG, kg/d      d 0 to 11  0.24  0.24  0.24  0.02      d 11 to 20  0.29  0.30  0.30  0.03      d 0 to 20  0.26  0.27  0.26  0.02  1Values are least squares means. 2Day after vitamin administration except d 0 is before administration. 3Control = no vitamin administration; Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. 4Actual mean day was 19.6 d after administration. View Large Table 2. Effect of administration routes of fat-soluble vitamins on growth performance for suckling pigs1   Treatment3    Day2  Control  Oral  Injection  SEM  BW, kg      d 0  1.87  1.85  1.88  0.07      d 11  4.52  4.50  4.49  0.24      d 204  7.03  7.09  7.06  0.46  ADG, kg/d      d 0 to 11  0.24  0.24  0.24  0.02      d 11 to 20  0.29  0.30  0.30  0.03      d 0 to 20  0.26  0.27  0.26  0.02    Treatment3    Day2  Control  Oral  Injection  SEM  BW, kg      d 0  1.87  1.85  1.88  0.07      d 11  4.52  4.50  4.49  0.24      d 204  7.03  7.09  7.06  0.46  ADG, kg/d      d 0 to 11  0.24  0.24  0.24  0.02      d 11 to 20  0.29  0.30  0.30  0.03      d 0 to 20  0.26  0.27  0.26  0.02  1Values are least squares means. 2Day after vitamin administration except d 0 is before administration. 3Control = no vitamin administration; Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. 4Actual mean day was 19.6 d after administration. View Large Table 3. Effect of administration routes of fat-soluble vitamins on 25-hydroxycholecalciferol concentrations (ng/mL) for suckling pigs1,2,3   Treatment5    Day4  Control  Oral  Injection  SEM  0  3.0  2.9  3.1  0.20  1  4.0c  138.8b  212.5a  18.33  2  4.6c  155.4b  235.4a  10.37  3  5.3c  145.9b  222.8a  8.55  4  5.5c  145.2b  203.6a  9.60  6  5.8c  108.2b  174.9a  7.68  9  5.8c  64.3b  115.4a  8.73  14  5.4c  33.2b  67.4a  5.40  20  5.7c  16.3b  39.2a  3.18    Treatment5    Day4  Control  Oral  Injection  SEM  0  3.0  2.9  3.1  0.20  1  4.0c  138.8b  212.5a  18.33  2  4.6c  155.4b  235.4a  10.37  3  5.3c  145.9b  222.8a  8.55  4  5.5c  145.2b  203.6a  9.60  6  5.8c  108.2b  174.9a  7.68  9  5.8c  64.3b  115.4a  8.73  14  5.4c  33.2b  67.4a  5.40  20  5.7c  16.3b  39.2a  3.18  a–cMeans within the same row without a common superscript differ (P < 0.05). 1Values are least squares means that represent data from 5 observations per treatment for each day with each observation a value for a pooled sample from 3 pigs. There was 1 value below the detection limit (on d 0) and 1 outlier (high, on d 20) in the control treatment. 2Repeated measures ANOVA (the treatment and day effects and day × treatment interaction, P < 0.01). 3To convert nanograms per milliliter to nanomoles per liter, multiply by 2.496. 4Day after vitamin administration except d 0 is before administration. 5Control = no vitamin administration; Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. View Large Table 3. Effect of administration routes of fat-soluble vitamins on 25-hydroxycholecalciferol concentrations (ng/mL) for suckling pigs1,2,3   Treatment5    Day4  Control  Oral  Injection  SEM  0  3.0  2.9  3.1  0.20  1  4.0c  138.8b  212.5a  18.33  2  4.6c  155.4b  235.4a  10.37  3  5.3c  145.9b  222.8a  8.55  4  5.5c  145.2b  203.6a  9.60  6  5.8c  108.2b  174.9a  7.68  9  5.8c  64.3b  115.4a  8.73  14  5.4c  33.2b  67.4a  5.40  20  5.7c  16.3b  39.2a  3.18    Treatment5    Day4  Control  Oral  Injection  SEM  0  3.0  2.9  3.1  0.20  1  4.0c  138.8b  212.5a  18.33  2  4.6c  155.4b  235.4a  10.37  3  5.3c  145.9b  222.8a  8.55  4  5.5c  145.2b  203.6a  9.60  6  5.8c  108.2b  174.9a  7.68  9  5.8c  64.3b  115.4a  8.73  14  5.4c  33.2b  67.4a  5.40  20  5.7c  16.3b  39.2a  3.18  a–cMeans within the same row without a common superscript differ (P < 0.05). 1Values are least squares means that represent data from 5 observations per treatment for each day with each observation a value for a pooled sample from 3 pigs. There was 1 value below the detection limit (on d 0) and 1 outlier (high, on d 20) in the control treatment. 2Repeated measures ANOVA (the treatment and day effects and day × treatment interaction, P < 0.01). 3To convert nanograms per milliliter to nanomoles per liter, multiply by 2.496. 4Day after vitamin administration except d 0 is before administration. 5Control = no vitamin administration; Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. View Large For plasma α-tocopherol concentrations, the effects of treatment, day, and day × treatment interaction were observed (P < 0.01; Table 4). Plasma α-tocopherol concentrations increased after administration to peak at d 1 after administration and then decreased thereafter. Pigs in the injection treatment had greater plasma α-tocopherol concentrations compared with those in the control and oral treatments from d 1 to 20 after administration (P < 0.05) except d 6 and 9 after administration. On d 6 after administration, the injection treatment was greater for plasma α-tocopherol concentrations than the control treatment but similar to the oral treatment (P < 0.05). Plasma α-tocopherol concentrations in the oral treatment were greater than those in the control treatment on d 1 (P < 0.05), with tendencies (P < 0.10) d 2, 3, and 6 after administration. Table 4. Effect of administration routes of fat-soluble vitamins on α-tocopherol concentrations (μg/mL) for suckling pigs1,2,3   Treatment5    Day4  Control  Oral  Injection  SEM  0  1.8  2.1  1.6  0.48  1  3.9c  14.3b  26.1a  2.30  2  6.7b,y  12.4b,x  23.6a,z  1.95  3  4.7b,y  10.0b,x  21.3a,z  2.05  4  4.7b  6.2b  17.9a  1.36  6  2.8b,y  7.4ab,x  10.4a,x  1.68  9  2.4  2.5  1.5  0.88  14  3.5b  3.3b  4.9a  0.39  20  4.1b  3.8b  5.7a  0.41    Treatment5    Day4  Control  Oral  Injection  SEM  0  1.8  2.1  1.6  0.48  1  3.9c  14.3b  26.1a  2.30  2  6.7b,y  12.4b,x  23.6a,z  1.95  3  4.7b,y  10.0b,x  21.3a,z  2.05  4  4.7b  6.2b  17.9a  1.36  6  2.8b,y  7.4ab,x  10.4a,x  1.68  9  2.4  2.5  1.5  0.88  14  3.5b  3.3b  4.9a  0.39  20  4.1b  3.8b  5.7a  0.41  a–cMeans within the same row without a common superscript differ (P < 0.05). x–zMeans within the same row without a common superscript differ (P < 0.10). 1Values are least squares means that represent data from 5 observations per treatment for each day with each observation a value for a pooled sample from 3 pigs. There are 6 (1 on d 0, 2 on d 2, 2 on d 4, and 1 on d 20), 2 (2 on d 6), and 3 (1 on d 0 and 2 on d 9) values below the detection limit in the control, oral, and injection treatments, respectively, and 2 outliers (1 on d 1, high; and 1 on d 4, low) in the injection treatment. 2Repeated measures ANOVA (the treatment and day effects and day × treatment interaction, P < 0.01). 3To convert micrograms per milliliter to micromoles per liter, multiply by 2.322. 4Day after vitamin administration except d 0 is before administration. 5Control = no vitamin administration; Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. View Large Table 4. Effect of administration routes of fat-soluble vitamins on α-tocopherol concentrations (μg/mL) for suckling pigs1,2,3   Treatment5    Day4  Control  Oral  Injection  SEM  0  1.8  2.1  1.6  0.48  1  3.9c  14.3b  26.1a  2.30  2  6.7b,y  12.4b,x  23.6a,z  1.95  3  4.7b,y  10.0b,x  21.3a,z  2.05  4  4.7b  6.2b  17.9a  1.36  6  2.8b,y  7.4ab,x  10.4a,x  1.68  9  2.4  2.5  1.5  0.88  14  3.5b  3.3b  4.9a  0.39  20  4.1b  3.8b  5.7a  0.41    Treatment5    Day4  Control  Oral  Injection  SEM  0  1.8  2.1  1.6  0.48  1  3.9c  14.3b  26.1a  2.30  2  6.7b,y  12.4b,x  23.6a,z  1.95  3  4.7b,y  10.0b,x  21.3a,z  2.05  4  4.7b  6.2b  17.9a  1.36  6  2.8b,y  7.4ab,x  10.4a,x  1.68  9  2.4  2.5  1.5  0.88  14  3.5b  3.3b  4.9a  0.39  20  4.1b  3.8b  5.7a  0.41  a–cMeans within the same row without a common superscript differ (P < 0.05). x–zMeans within the same row without a common superscript differ (P < 0.10). 1Values are least squares means that represent data from 5 observations per treatment for each day with each observation a value for a pooled sample from 3 pigs. There are 6 (1 on d 0, 2 on d 2, 2 on d 4, and 1 on d 20), 2 (2 on d 6), and 3 (1 on d 0 and 2 on d 9) values below the detection limit in the control, oral, and injection treatments, respectively, and 2 outliers (1 on d 1, high; and 1 on d 4, low) in the injection treatment. 2Repeated measures ANOVA (the treatment and day effects and day × treatment interaction, P < 0.01). 3To convert micrograms per milliliter to micromoles per liter, multiply by 2.322. 4Day after vitamin administration except d 0 is before administration. 5Control = no vitamin administration; Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. View Large For plasma concentrations of retinyl palmitate (the form of vitamin A administered), there were also the effects of treatment, day, and day × treatment interaction (P < 0.01; Table 5). After administration, plasma retinyl palmitate concentrations in the injection treatment increased to reach peak at d 1 of after administration and then decreased thereafter. Pigs injected with retinyl palmitate had greater plasma values than those in the control and oral treatments on d 1 to 4 after administration (P < 0.05) whereas plasma values in the oral treatment did not differ from those in the control treatment, even though plasma value almost doubled at d 1 after administration. From d 6 after administration, there was no significant difference in plasma retinyl palmitate concentrations among the treatments. Table 5. Effect of administration routes of fat-soluble vitamins on retinyl palmitate concentrations (μg/mL) for suckling pigs1,2,3   Treatment5    Day4  Control  Oral  Injection  SEM  0  0.12  0.14  0.15  0.015  1  0.13b  0.25b  2.17a  0.208  2  0.12b  0.12b  0.60a  0.049  3  0.07b  0.08b  0.37a  0.043  4  0.05b  0.06b  0.14a  0.023  6  0.05  0.04  0.12  0.042  9  0.08  0.04  0.02  0.020  14  0.03  0.03  0.03  0.002  20  0.03  0.03  0.03  0.003    Treatment5    Day4  Control  Oral  Injection  SEM  0  0.12  0.14  0.15  0.015  1  0.13b  0.25b  2.17a  0.208  2  0.12b  0.12b  0.60a  0.049  3  0.07b  0.08b  0.37a  0.043  4  0.05b  0.06b  0.14a  0.023  6  0.05  0.04  0.12  0.042  9  0.08  0.04  0.02  0.020  14  0.03  0.03  0.03  0.002  20  0.03  0.03  0.03  0.003  a,bMeans within the same row without a common superscript differ (P < 0.05). 1Values are least squares means that represent data from 5 observations per treatment for each day with each observation a value for a pooled sample from 3 pigs. There are 9 (2 on d 3, 2 on d 4, 2 on d 6, 2 on d 9, and 1 on d 14) and 6 (1 on d 2, 1 on d 4, 3 on d 9, and 1 on d 14) values below the detection limit in the control and oral treatments, respectively, and 1 (on d 1, high) and 3 (1 on d 0, low; 1 on d 1, high; and 1 on d 2, high) outliers in the oral and injection treatments, respectively. 2Repeated measures ANOVA (the treatment and day effects and day × treatment interaction, P < 0.01). 3To convert micrograms per milliliter to micromoles per liter, multiply by 3.491. 4Day after vitamin administration except d 0 is before administration. 5Control = no vitamin administration; Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. View Large Table 5. Effect of administration routes of fat-soluble vitamins on retinyl palmitate concentrations (μg/mL) for suckling pigs1,2,3   Treatment5    Day4  Control  Oral  Injection  SEM  0  0.12  0.14  0.15  0.015  1  0.13b  0.25b  2.17a  0.208  2  0.12b  0.12b  0.60a  0.049  3  0.07b  0.08b  0.37a  0.043  4  0.05b  0.06b  0.14a  0.023  6  0.05  0.04  0.12  0.042  9  0.08  0.04  0.02  0.020  14  0.03  0.03  0.03  0.002  20  0.03  0.03  0.03  0.003    Treatment5    Day4  Control  Oral  Injection  SEM  0  0.12  0.14  0.15  0.015  1  0.13b  0.25b  2.17a  0.208  2  0.12b  0.12b  0.60a  0.049  3  0.07b  0.08b  0.37a  0.043  4  0.05b  0.06b  0.14a  0.023  6  0.05  0.04  0.12  0.042  9  0.08  0.04  0.02  0.020  14  0.03  0.03  0.03  0.002  20  0.03  0.03  0.03  0.003  a,bMeans within the same row without a common superscript differ (P < 0.05). 1Values are least squares means that represent data from 5 observations per treatment for each day with each observation a value for a pooled sample from 3 pigs. There are 9 (2 on d 3, 2 on d 4, 2 on d 6, 2 on d 9, and 1 on d 14) and 6 (1 on d 2, 1 on d 4, 3 on d 9, and 1 on d 14) values below the detection limit in the control and oral treatments, respectively, and 1 (on d 1, high) and 3 (1 on d 0, low; 1 on d 1, high; and 1 on d 2, high) outliers in the oral and injection treatments, respectively. 2Repeated measures ANOVA (the treatment and day effects and day × treatment interaction, P < 0.01). 3To convert micrograms per milliliter to micromoles per liter, multiply by 3.491. 4Day after vitamin administration except d 0 is before administration. 5Control = no vitamin administration; Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. View Large Unlike the results of plasma retinyl palmitate concentrations, plasma retinol concentrations did not differ by retinyl palmitate administration (Table 6). There was only the day effect, in which plasma retinol concentrations on d 0 were lower than d 1 after administration and later (P < 0.05; d 2 after administration, P = 0.06) and that on d 2 was lower than d 6 after administration and later (P < 0.05). Table 6. Effect of administration routes of fat-soluble vitamins on retinol concentrations (μg/mL) for suckling pigs1,2,3   Treatment5    Day4  Control  Oral  Injection  SEM  0  0.07  0.09  0.09  0.005  1  0.12  0.12  0.14  0.008  2  0.09  0.08  0.12  0.015  3  0.12  0.12  0.14  0.020  4  0.11  0.10  0.14  0.015  6  0.13  0.18  0.14  0.018  9  0.13  0.16  0.14  0.022  14  0.12  0.12  0.13  0.009  20  0.14  0.15  0.15  0.013    Treatment5    Day4  Control  Oral  Injection  SEM  0  0.07  0.09  0.09  0.005  1  0.12  0.12  0.14  0.008  2  0.09  0.08  0.12  0.015  3  0.12  0.12  0.14  0.020  4  0.11  0.10  0.14  0.015  6  0.13  0.18  0.14  0.018  9  0.13  0.16  0.14  0.022  14  0.12  0.12  0.13  0.009  20  0.14  0.15  0.15  0.013  1Values are least squares means that represent data from 5 observations per treatment for each day with each observation a value for a pooled sample from 3 pigs with no missing values. 2Repeated measures ANOVA (the day effect, P < 0.01; no effects of treatment and day × treatment interaction). 3To convert micrograms per milliliter to micromoles per liter, multiply by 3.491. 4Day after vitamin administration except d 0 is before administration. 5Control = no vitamin administration; Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. View Large Table 6. Effect of administration routes of fat-soluble vitamins on retinol concentrations (μg/mL) for suckling pigs1,2,3   Treatment5    Day4  Control  Oral  Injection  SEM  0  0.07  0.09  0.09  0.005  1  0.12  0.12  0.14  0.008  2  0.09  0.08  0.12  0.015  3  0.12  0.12  0.14  0.020  4  0.11  0.10  0.14  0.015  6  0.13  0.18  0.14  0.018  9  0.13  0.16  0.14  0.022  14  0.12  0.12  0.13  0.009  20  0.14  0.15  0.15  0.013    Treatment5    Day4  Control  Oral  Injection  SEM  0  0.07  0.09  0.09  0.005  1  0.12  0.12  0.14  0.008  2  0.09  0.08  0.12  0.015  3  0.12  0.12  0.14  0.020  4  0.11  0.10  0.14  0.015  6  0.13  0.18  0.14  0.018  9  0.13  0.16  0.14  0.022  14  0.12  0.12  0.13  0.009  20  0.14  0.15  0.15  0.013  1Values are least squares means that represent data from 5 observations per treatment for each day with each observation a value for a pooled sample from 3 pigs with no missing values. 2Repeated measures ANOVA (the day effect, P < 0.01; no effects of treatment and day × treatment interaction). 3To convert micrograms per milliliter to micromoles per liter, multiply by 3.491. 4Day after vitamin administration except d 0 is before administration. 5Control = no vitamin administration; Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. View Large In the plasma vitamin concentrations of the control pigs, plasma 25OHD3 (Fig. 1a) and retinol (Fig. 1c) concentrations increased within the first week of birth and were maintained constantly until weaning whereas plasma α-tocopherol (Fig. 1b) and retinyl palmitate (Fig. 1c) concentrations increased from birth, peaked at d 3 and 2 of age, respectively, and then decreased afterward until weaning. Figure 1. View largeDownload slide Time-dependent plasma 25-hydroxycholecalciferol (25OHD3; a), α-tocopherol (b), and retinyl palmitate and retinol (c) concentrations for control pigs. Values are means ± SEM. To convert nanograms per milliliter to nanomoles per liter for 25OHD3, multiply by 2.496. To convert micrograms per milliliter to micromoles per liter for α-tocopherol, multiply by 2.322. To convert micrograms per milliliter to micromoles per liter for retinyl palmitate and retinol, multiply by 3.491. Figure 1. View largeDownload slide Time-dependent plasma 25-hydroxycholecalciferol (25OHD3; a), α-tocopherol (b), and retinyl palmitate and retinol (c) concentrations for control pigs. Values are means ± SEM. To convert nanograms per milliliter to nanomoles per liter for 25OHD3, multiply by 2.496. To convert micrograms per milliliter to micromoles per liter for α-tocopherol, multiply by 2.322. To convert micrograms per milliliter to micromoles per liter for retinyl palmitate and retinol, multiply by 3.491. Based on time-dependent plasma concentration profiles for 25OHD3 (Fig. 2), pharmacokinetic parameters were obtained (Table 7). In the results of pharmacokinetic parameters for plasma 25OHD3 concentrations, the injection treatment had greater tel1/2 (P < 0.01), Cmax (P < 0.05), and all of the AUC parameters (P < 0.01) but lower kel (P < 0.01) than the oral treatment. The values of ka, ta1/2, and Tmax were not different between the injection and oral treatments. The relative bioavailability of oral administration compared with injection administration had a mean of 55.26% (SEM 4.28). The pharmacokinetic parameters for α-tocopherol, retinyl palmitate, and retinol could not be obtained by the pharmacokinetic model used for plasma 25OHD3 concentrations due to the presence of missing values for some litters for certain sampling time points monitored in conjunction with different shapes of the response curves. It was not possible to obtain the AUC0–∞ for plasma α-tocopherol concentrations that is required for the pharmacokinetic model used for plasma 25OHD3 concentrations in the current study. However, in the simple calculation of the AUC for days 0 through 20 after administration for α-tocopherol using the overall treatment mean values, a relative bioavailability estimate of 60.27% for oral administration compared with injection administration is obtained. Figure 2. View largeDownload slide The plasma concentration–time profile of 25-hydroxycholecalciferol (25OHD3) concentrations in pigs. Values are means ± SEM. Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. To convert nanograms per milliliter to nanomoles per liter, multiply by 2.496. Figure 2. View largeDownload slide The plasma concentration–time profile of 25-hydroxycholecalciferol (25OHD3) concentrations in pigs. Values are means ± SEM. Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. To convert nanograms per milliliter to nanomoles per liter, multiply by 2.496. Table 7. Pharmacokinetics of plasma 25-hydroxycholecalciferol concentrations in pigs1   Treatment3      Item2  Oral  Injection  SEM  P-value  ka, d–1  1.154  1.608  0.2417  0.255  kel, d–1  0.148  0.112  0.0075  0.003  ta1/2, d  0.659  0.498  0.0991  0.316  tel1/2, d  4.76  6.33  0.389  0.006  Tmax, d  2.137  1.934  0.2251  0.557  Cmax, ng/mL4  161.0  240.7  13.76  0.010  AUC0–20, ng/mL∙d  1,418.5  2,364.8  166.62  0.001  AUC20–∞, ng/mL∙d  115.7  369.4  59.33  0.009  AUC0–∞, ng/mL∙d  1,534.3  2,734.2  214.36  <0.001    Treatment3      Item2  Oral  Injection  SEM  P-value  ka, d–1  1.154  1.608  0.2417  0.255  kel, d–1  0.148  0.112  0.0075  0.003  ta1/2, d  0.659  0.498  0.0991  0.316  tel1/2, d  4.76  6.33  0.389  0.006  Tmax, d  2.137  1.934  0.2251  0.557  Cmax, ng/mL4  161.0  240.7  13.76  0.010  AUC0–20, ng/mL∙d  1,418.5  2,364.8  166.62  0.001  AUC20–∞, ng/mL∙d  115.7  369.4  59.33  0.009  AUC0–∞, ng/mL∙d  1,534.3  2,734.2  214.36  <0.001  1Values are least squares means that represent data from 5 independently computed curves per treatment. 2ka = the absorption rate constant; kel = the elimination rate constant; ta1/2 = the half-life of absorption; tel1/2 = the half-life of elimination; Tmax = the time to reach maximum concentration; Cmax = the maximum concentration; AUC0–20 = the area under the curve (AUC) from d 0 to the last sampling day; AUC20–∞ = the extrapolated AUC from the last sampling day to infinity; AUC0–∞ = the AUC from d 0 to infinity. 3Control = no vitamin administration; Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. 4To convert nanograms per milliliter to nanomoles per liter, multiply by 2.496. View Large Table 7. Pharmacokinetics of plasma 25-hydroxycholecalciferol concentrations in pigs1   Treatment3      Item2  Oral  Injection  SEM  P-value  ka, d–1  1.154  1.608  0.2417  0.255  kel, d–1  0.148  0.112  0.0075  0.003  ta1/2, d  0.659  0.498  0.0991  0.316  tel1/2, d  4.76  6.33  0.389  0.006  Tmax, d  2.137  1.934  0.2251  0.557  Cmax, ng/mL4  161.0  240.7  13.76  0.010  AUC0–20, ng/mL∙d  1,418.5  2,364.8  166.62  0.001  AUC20–∞, ng/mL∙d  115.7  369.4  59.33  0.009  AUC0–∞, ng/mL∙d  1,534.3  2,734.2  214.36  <0.001    Treatment3      Item2  Oral  Injection  SEM  P-value  ka, d–1  1.154  1.608  0.2417  0.255  kel, d–1  0.148  0.112  0.0075  0.003  ta1/2, d  0.659  0.498  0.0991  0.316  tel1/2, d  4.76  6.33  0.389  0.006  Tmax, d  2.137  1.934  0.2251  0.557  Cmax, ng/mL4  161.0  240.7  13.76  0.010  AUC0–20, ng/mL∙d  1,418.5  2,364.8  166.62  0.001  AUC20–∞, ng/mL∙d  115.7  369.4  59.33  0.009  AUC0–∞, ng/mL∙d  1,534.3  2,734.2  214.36  <0.001  1Values are least squares means that represent data from 5 independently computed curves per treatment. 2ka = the absorption rate constant; kel = the elimination rate constant; ta1/2 = the half-life of absorption; tel1/2 = the half-life of elimination; Tmax = the time to reach maximum concentration; Cmax = the maximum concentration; AUC0–20 = the area under the curve (AUC) from d 0 to the last sampling day; AUC20–∞ = the extrapolated AUC from the last sampling day to infinity; AUC0–∞ = the AUC from d 0 to infinity. 3Control = no vitamin administration; Oral = oral administration with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol; Injection = i.m. injection with 40,000 IU of vitamin D3, 40,000 IU of vitamin A as retinyl palmitate, and 400 IU of vitamin E as D-α-tocopherol. 4To convert nanograms per milliliter to nanomoles per liter, multiply by 2.496. View Large DISCUSSION The current study evaluated whether fat-soluble vitamin administration to newborn pigs via different administration routes affected the temporal profile of plasma vitamin status during the suckling period. Because of the physiological similarity of pigs and humans (Swindle et al., 2012), the current study can represent nutritional aspects of fat-soluble vitamins for humans. Recent human studies have reported that vitamin D status influences not only Ca homeostasis and bone health but also diseases such as inflammatory bowel disease, respiratory infection, anemia, and autism (Kong et al., 2008; Camargo et al., 2011; Atkinson et al., 2014; Patrick and Ames, 2014). Similarly to pigs, human breast milk reportedly does not contain sufficient amounts of vitamin A, D, and E (Greer, 2001), so it is not surprising that newborn and nursing infants are susceptible to vitamin deficiency. Specifically regarding vitamin D status, because of a low amount of vitamin D in breast milk and lack of exposure to sunlight, breastfed infants are vulnerable to vitamin D deficiency (Kaludjerovic and Vieth, 2010; Kovacs, 2012). Additionally, it is suggested that a low serum 25OHD3 concentration for infants could impair their development and subsequently affect adult health (Kaludjerovic and Vieth, 2010). Therefore, vitamin D supplementation for infants is suggested to start at birth (Ziegler et al., 2014) and may need to be increased to levels some have previously thought unsafe (Gallo et al., 2013). Many efforts have been conducted to improve fat-soluble vitamin status for nursing infants (Henriksen et al., 2006; Wagner et al., 2010; Shneider et al., 2012; Gallo et al., 2013) and demonstrated that daily oral supplementation of these vitamins could increase plasma vitamin levels. However, there is limited information on the temporal change of plasma vitamin status in human infant after fat-soluble vitamin supplementation/administration. Therefore, through the investigation on temporal profile of plasma vitamin status in suckling pigs, this study can contribute meaningful data for potential use in a nutritional area of intense current interest in human health. In the current study, the fat-soluble vitamin administration did not affect growth performance of the suckling piglets, which agrees with our previous study (Jang et al., 2014). The effects of fat-soluble vitamin administration/supplementation to pigs have been reported to improve vitamin status and prevent vitamin deficiency (Ching et al., 2002; Surles et al., 2007; Flohr et al., 2014; Jang et al., 2014). In the current study, the administration of vitamin D3, α-tocopherol, and retinyl palmitate to pigs at birth changed plasma status of 25OHD3, α-tocopherol, and retinyl palmitate but not retinol and the interaction between day and treatment demonstrated different patterns in the change of plasma vitamin concentrations by day between the types of vitamin administered and between administration routes. The peak day of plasma concentrations of α-tocopherol in both administration routes and retinyl palmitate in the injection route was 1 d earlier compared with that of plasma 25OHD3 concentrations. After the peak, plasma levels of these vitamins decreased progressively. However, there were differences in rates of reductions between types of vitamins, in which plasma retinyl palmitate concentrations decreased to be similar with the control treatment at d 6 after administration, which was faster than the reductions of plasma 25OHD3 and α-tocopherol concentrations. There is no study available that has investigated the time-dependent plasma vitamin profiles for vitamin A, D3, and E in pigs. In other species, plasma 25OHD3 concentrations peaked at 48 to 96 h after vitamin D3 administration by a bolus for calves with average 100 kg BW (Sommerfeldt et al., 1983). Ilahi et al. (2008) reported from a human study with healthy adults (average 71 kg BW) that plasma 25OHD3 concentrations reached the maximum level at d 7 after a single oral dose of 100,000 IU vitamin D3 and then decreased to reach the baseline at d 84 after administration. Regarding vitamin E administration, plasma α-tocopherol concentrations were maximized at 12 to 24 h after administration and then decreased to reach the baseline at d 3 after i.m. injection of 500 IU DL-α-tocopherol for sheep weighing 32 to 47 kg (Njeru et al., 1994). Kaseki et al. (1986) also reported that plasma α-tocopherol concentrations were maximized at 12 h after a single oral administration of 3 to 25 IU D-α-tocopherol for vitamin E–deficient rats. In the case of vitamin A, when 10,000 IU of retinyl palmitate was administered to dogs weighing 10 kg by a single oral dose, the peak of plasma retinyl palmitate concentrations were observed at 8 h after administration and then decreased to the baseline by 72 h after administration (Raila et al., 2002). In humans, plasma retinyl palmitate concentrations peaked at 3.5 to 12 h after a single oral dose of 100,000 IU retinyl palmitate to healthy male adults and then decreased to the baseline by 24 h after administration (Reinersdorff et al., 1996). The response to vitamin administration varies by vitamin dose administered and body size, and changes in plasma values after administration may be associated with the relative growth rate. Because preweaning relative growth rate is faster in pigs than lambs, calves, and infants (Miller and Ullrey, 1997), it was suggested (Jang et al., 2014) that the response of newborn pigs to fat-soluble vitamin administration may be a more rapid “dilution” related to the increase in blood volume associated with the relative increase in body mass. Collectively, in those previous studies, the maximum plasma concentrations of α-tocopherol and retinyl palmitate were observed at 12 to 24 h and 3 to 12 h, respectively. Unlike α-tocopherol and retinyl palmitate, plasma 25OHD3 concentrations peaked at d 2 to 7 after administration of vitamin D3. This differential response between the types of vitamins administered may be attributed to the metabolism (absorption/clearance) of each vitamin in the animal body and the vitamin forms analyzed. For vitamin D3, 25OHD3 is a circulating form in blood converted from vitamin D3 in the liver (Dittmer and Thompson, 2011). This could be a possible explanation why plasma 25OHD3 concentrations were maximized with a further increase that occurred from d 1 to 2 after administration. Sommerfeldt et al. (1983) reported when the radioactive-labeled vitamin D3 was orally administered to dairy calves, plasma levels of labeled vitamin D3 were predominant, peaked between 24 and 48 h, and then disappeared within 7.5 d, whereas plasma levels of labeled 25OHD3 became predominant, reached their maximum concentrations between 48 and 96 h, and then decreased afterward but still maintained greater levels than those at the initial point until d 24 after administration. Conversely, α-tocopherol is absorbed as a free alcohol form, transported to the liver complexed in a chylomicron, and released into blood as α-tocopherol–lipoprotein complex (Bjørneboe et al., 1990). Traber et al. (1990) reported, from the human study that analyzed labeled α-tocopherol concentrations in chylomicron, lipoprotein, and plasma after a single oral dose of labeled α-tocopheryl acetate, that labeled α-tocopherol concentrations in the plasma of control subjects peaked after being maximized in the chylomicrons but before being maximized in the lipoproteins. In case of retinyl palmitate, it is transported to the liver complexed into chylomicron, stored in the liver as an ester form, and released into blood as an alcohol form (retinol) combined with retinol-binding protein (Harrison and Hussain, 2001). These indicate that plasma retinyl palmitate concentrations analyzed herein may represent the appearance and clearance of retinyl palmitate administered itself whereas plasma retinol concentrations analyzed herein may represent the metabolized and released form from the liver. Therefore, it could be suggested that the disappearance of plasma retinyl palmitate was faster than that of plasma α-tocopherol and that the disappearance of plasma 25OHD3 was slower than those of plasma α-tocopherol and retinyl palmitate. Additionally, it may be noted that the plasma values of α-tocopherol and retinyl palmitate observed as maximum values might not be the peak values that fully responded to the administration because only 1 sampling point per day existed for absorption phase by vitamin administration of α-tocopherol and retinyl palmitate. Regarding plasma retinol concentrations, the plasma level was not affected by retinyl palmitate administration. Because the liver stores a large amount of vitamin A as retinyl palmitate and releases free retinol when needed, it regulates the release of retinol into the blood and there is low correlation between liver and blood retinol concentrations (Underwood et al., 1979; Harrison and Hussain, 2001). Therefore, even though plasma retinyl palmitate concentrations increased after administration, plasma retinol concentrations may be under the regulation by the liver resulting in no effect of retinyl palmitate administration in plasma retinol concentrations regardless of administration routes. However, there was a day effect in which plasma retinol concentrations were greater on d 1 after administration and later compared with the initial value and increased within the first week of birth. This result may be attributed to continuous vitamin A ingestion of suckling pigs via colostrum and milk. Håkansson et al. (2001) reported that sow colostrum and milk contained 2.5 and 0.6 to 0.7 μg/mL of retinol, respectively, and that plasma retinol concentrations increased continuously from birth. In the comparison of administration routes, the plasma levels of fat-soluble vitamins increased more efficiently and decreased more slowly by the injection administration than the oral administration. These results may be attributed to the process of absorption after vitamin administration. Because oral administration of vitamins must undergo the processes of the digestive tract, all vitamins administered may not be entirely absorbed (McDowell, 2000). However, the injected vitamins could not be excreted unless fully metabolized so that the injected amount should be circulated or stored in the body until cleared. Nevertheless, after peaking, plasma vitamin concentrations were continuously reduced until weaning. Therefore, continuous administration/supplementation of these vitamins might be needed to sustain high plasma values. Data from the control treatment showed the normal change of plasma vitamin concentrations in pigs that consumed only milk from sows. Plasma 25OHD3, α-tocopherol, retinyl palmitate, and retinol concentrations increased within the first week of birth and peaked at d 7, 3, 2, and 7 of age, respectively. However, plasma 25OHD3 and retinol concentrations seemed to be maintained constantly thereafter until weaning whereas plasma α-tocopherol and retinyl palmitate concentrations decreased thereafter. These results of the control pigs closely agree with previous studies (Håkansson et al., 2001; Jang et al., 2014). As mentioned above, newborn pigs grow rapidly, which results in approximately 4 times greater BW at weaning at about d 21 of age compared with that at birth in this study. Because blood volume is linearly increased with increasing BW (Doornenbal and Martin, 1965), an increase or maintenance of a plasma constituent can occur only if milk contains an appreciable amount of the constituent. A reduction of plasma vitamin concentrations in the control pigs after the first week of age might be attributed to both a dilution effect by the increase in blood volume and by normal declines associated with the metabolic use of the vitamin. Additionally, the greatest increases appeared between d 1 and 2 of age in plasma 25OHD3 and retinol concentrations and between d 2 and 3 of age in plasma α-tocopherol concentrations. This result reflects the contribution of colostrum that contains high amount of these vitamins (Goff et al., 1984; Håkansson et al., 2001). Therefore, these results mean that colostrum contributes plasma vitamin concentrations for suckling piglets for early lactation period but it does not lead to continuous increase of plasma vitamin levels. This is the first study that investigated the effect of administration routes of vitamin D3 on pharmacokinetics in plasma 25OHD3 concentrations for pigs to demonstrate the different responses after administration between oral and injection treatments. The plasma 25OHD3 concentrations were changed biexponentially over time after administration with an increase immediately after administration until a peak and a decrease afterward. This biexponential change agrees with previous studies conducted in humans (Ilahi et al., 2008; Jones et al., 2012), rats (Brouwer et al., 1998), and calves (Sommerfeldt et al., 1983). Pharmacokinetic parameters are affected by several factors such as species (human, rat, pig, sheep, etc.), vitamin sources, administration routes, dose, and duration (Whyte et al., 1979; Horst et al., 1982; Sommerfeldt et al., 1983, Toutain et al., 1992). In this study, Tmax in both oral and injection treatments occurred at approximately d 2 and agrees with the results from calf (Sommerfeldt et al., 1983) and rat (Brouwer et al., 1998) studies whereas it was largely different from the human (Ilahi et al., 2008). Pharmacokinetic analysis for oral and injection treatments demonstrated that the parameters for distribution of 25OHD3 (i.e., ka, ta1/2, and Tmax) in the oral treatment did not differ from the injection treatment, indicating that the distribution responses of pigs for 25OHD3 were similar between the 2 administration routes. However, in the elimination phase, the reduction of plasma 25OHD3 concentrations of pigs were slower in the injection treatment than the oral treatment, as shown by lower kel and longer tel1/2 in the injection treatment than the oral treatment. Also, the injection treatment had greater Cmax and AUC parameters (AUC0–20, AUC20–∞, and AUC0–∞) than the oral treatment, resulting in a relative bioavailability estimate of 55.26% for oral administration compared with injection administration. Whyte et al. (1979) reported that vitamin D bioavailability varied by administration routes (i.e., oral, subcutaneous, i.m., and i.v.) of vitamin D2 or D3 and that i.m. injection increased plasma 25OHD3 concentrations greater than oral administration when 240,000 IU of vitamin D3 was administered to normal adults. Vieth (1999) reported that a single i.m. dose of vitamin D had a longer-lasting response than an oral dose on serum 25OHD3 concentrations in human. Therefore, these results mean that the injectable route of vitamin D3 lead to a greater increase and a slower reduction in plasma 25OHD3 concentrations than the oral route for suckling piglets, which agrees with Jang et al. (2014) even though plasma 25OHD3 distribution rate after vitamin D3 administration were similar regardless of administration routes. In conclusion, fat-soluble vitamin administration to newborn pigs changed the plasma status of 25OHD3, α-tocopherol, and retinyl palmitate immediately after administration and the plasma profile changes of these vitamins were different between administration routes. The response to vitamin administration was faster in plasma retinyl palmitate than in plasma 25OHD3 and α-tocopherol. The injection administration resulted in the greater increase and slower disappearance of plasma vitamin levels than the oral administration during the suckling period. Additionally, the injection administration increased plasma 25OHD3, α-tocopherol, and retinyl palmitate concentrations whereas the oral administration increased only plasma 25OHD3 and α-tocopherol concentrations but not plasma retinyl palmitate concentrations. The greatest increases of plasma 25OHD3, α-tocopherol, and retinol concentrations in the control pigs occurred between d 1 and 3 of age. 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TI - Temporal plasma vitamin concentrations are altered by fat-soluble vitamin administration in suckling pigs
JF - Journal of Animal Science
DO - 10.2527/jas.2015-9221
DA - 2015-11-01
UR - https://www.deepdyve.com/lp/oxford-university-press/temporal-plasma-vitamin-concentrations-are-altered-by-fat-soluble-YzIP5XOa6t
SP - 5273
EP - 5282
VL - 93
IS - 11
DP - DeepDyve
ER -