Moderate Perinatal Choline Deficiency Elicits Altered Physiology and Metabolomic Profiles in the Piglet

<div><p>Few studies have evaluated the impact of dietary choline on the health and well-being of swine, and those pivotal papers were aimed at determining dietary requirements for sows and growing pigs. This is of importance as the piglet is becoming a widely accepted model for human infant nutrition, but little is known about the impacts of perinatal choline status on overall health and metabolism of the growing piglet. In the present study, sows were provided either a choline deficient (CD, 625 mg choline/kg dry matter) or choline sufficient (CS, 1306 mg choline/kg dry matter) diet for the last 65 d of gestation (prenatal intervention). Piglets were weaned from the sow 48 h after farrowing and provided either a CD (477 mg choline/kg dry matter) or CS (1528 mg choline/kg dry matter) milk replacer (postnatal intervention) for 29 ± 2 d, resulting in a factorial arrangement of 4 treatment (prenatal/postnatal) groups: CS/CS, CS/CD, CD/CS, and CD/CD. Piglet growth was normal for artificially-reared piglets, and was not impacted by perinatal choline status. Piglets receiving the postnatal CD treatment had lower (<i>P</i> < 0.01) plasma choline and choline-containing phospholipid concentrations and higher (<i>P</i> < 0.05) liver enzyme (alkaline phosphatase and gamma-glutamyl transferase) values compared with piglets receiving the postnatal CS treatment. Hepatic lipid content of piglets receiving the postnatal CD treatment was higher (<i>P</i> < 0.01) compared with piglets receiving the postnatal CS treatment. Additionally, postnatally CD piglets had lower (<i>P</i> = 0.01) plasma cholesterol than postnatally CS piglets. Brain development was also impacted by perinatal choline status, with brains of piglets exposed to prenatal CD being smaller (<i>P</i> = 0.01) than those of prenatally CS piglets. These findings support the hypothesis that the piglet is a sensitive model for choline deficiency during the perinatal period. In the present study, piglets exhibited similarities in health markers and metabolomic profiles to rodents and humans when exposed to moderate choline deficiency.</p></div

Effects of perinatal choline status on metabolomic profiles of 4-wk-old piglets¹.

<p>^abcMeans within a row and without a common superscript differ (<i>P</i> < 0.05).</p><p>¹Values are means of 8 replicate pigs exposed to prenatal and postnatal choline treatments (e.g., CS/CS as the control group) with blood collected from piglets at 27–30 d of age. Data presented as fold-change relative CS/CS treatment group. CD, choline deficient; CS, choline sufficient.</p><p>²Pre, main effect of prenatal choline status; Post, main effect of postnatal choline status; Pre x Post, interactive effect of prenatal and postnatal choline statuses.</p><p>Effects of perinatal choline status on metabolomic profiles of 4-wk-old piglets<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133500#t005fn002" target="_blank">¹</a>.</p

Effects of perinatal choline status on final body weights and brain size of piglets.

<p>Values are means of 8 replicate pigs exposed to prenatal and postnatal choline treatments (e.g., CS/CS as the control group). A) Piglet weights at 27 d of age did not differ between treatments. B) Regardless of postnatal choline status, pigs exposed to prenatal CD environment had smaller (<i>P</i> = 0.01) brains than those piglets exposed to a prenatal CS environment. Abbreviations: CD, choline deficient; CS, choline sufficient</p

Composition of gestation diet (choline-deficient)¹.

<p>¹Choline sufficient diet was produced by replacing 1.8 g of cornstarch per kg of final diet with choline chloride (containing 60% choline). Choline deficient and sufficient gestation diets were analyzed to contain 625 and 1,306 mg of choline per kg diet, respectively.</p><p>²Ardex F, Archer Daniels Midland, Decatur, IL.</p><p>³Provided per kg of complete diet: Ca, 0.6 mg (CaCO₃); P, 298.8 mg; Mg, 1.8 mg; K, 2.4 mg; Na, 0.3 mg; S, 172.2 mg; Zn, 125.1 mg; Fe, 129.1 mg (FeSO₄); Mn, 60.3 mg (MnSO₄); Cu, 10.2 mg (CuCl₂, CuSO₄); I, 1.3 mg; Se, 0.3 mg (Na₂SeO₃); Cl, 2.1 mg; Vit A, 11.1 kIU; Vit D, 2.2 kIU; Vit E, 66.1 kIU; Vit K, 1.4 mg (menadione dimethylpyrimidinol bisulfite); thiamin, 0.2 mg (thiamine mononitrate); riboflavin, 6.6 mg; niacin, 44.1 mg; pantothenic acid, 23.5 mg (D-calcium pantothenate); pyridoxine, 0.2 mg (pyridoxine-HCl); biotin, 0.4 mg; folacin, 1.6 mg; Vit B12, 0.03 mg.</p><p>Composition of gestation diet (choline-deficient)<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133500#t001fn001" target="_blank">¹</a>.</p

Composition of milk replacer formulation (choline deficient)¹.

<p>¹Choline sufficient milk replacer was produced by replacing 2.2 g lactose and 0.22g calcium chloride with choline chloride (containing 70% choline). Choline deficient and sufficient milk replacer formulations were analyzed to contain 477 and 1,528 mg of choline per kg diet, respectively.</p><p>²Ho-Milc 7–60, Merrick’s, Inc., Middleton, WI.</p><p>³Ardex F, Archer Daniels Midland, Decatur, IL.</p><p>⁴Provided per kg of complete diet: Cu, 23.5 mg (CuSO₄); Zn, 257.2 mg (ZnSO₄); Se, 1.0 mg (Na₂SeO₃); Mg, 2.88 g (MgCO₃); NaPO₄, 5.2 g; menadione, 5.1 mg; Vit C, 51.0 mg; Vit A, 47.0 mg; Vit D, 918.6 mg; Vit E, 673.6 mg; thiamin, 29.6 mg (10%); riboflavin, 102.1 mg; niacin, 62.3 mg (98%); biotin, 316.4 mg; Ca-pantothenate, 188.8 mg; pyridoxine, 306.2 mg; folate, 47.0 mg; Vit B12, 170.4 mg.</p><p>⁵Solka Floc, International Fiber Corporation, North Tonawanda, NY.</p><p>⁶Luctarom Milky Vanilla, Lucta USA, Inc., Northbrook, IL.</p><p>Composition of milk replacer formulation (choline deficient)<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133500#t002fn001" target="_blank">¹</a>.</p

Effects of perinatal choline status on relative concentrations of metabolites related to methyl metabolism in 4-wk-old piglets.

<p>Values are means of 8 replicate pigs exposed to prenatal and postnatal choline treatments (e.g., CS/CS as the control group) with blood samples collected at 27–30 d of age. Regardless of prenatal choline status, pigs fed a CD milk replacer exhibited lower (<i>P</i> < 0.01) plasma concentrations of: A) total choline + choline-containing phospholipids, B) choline, and C) betaine, compared with pigs fed CS milk replacer. D) Plasma methionine in piglets exposed to prenatal CD environment was lower (<i>P</i> = 0.05) compared with piglets exposed to a prenatal CS environment, regardless of postnatal choline status. Abbreviations: CD, choline deficient; CS, choline sufficient</p

Effects of perinatal choline status on clinical blood chemistry profiles of 4-wk-old piglets¹.

<p>^abMeans within a row and without a common superscript differ (<i>P</i> < 0.05).</p><p>¹Values are means of 8 replicate pigs exposed to prenatal and postnatal choline treatments (e.g., CS/CS as the control group) with blood collected from piglets at 27–30 d of age. ALP, alkaline phosphatase; AST, aspartate aminotransferase; BUN, blood urea nitrogen; CD, choline deficient; CS, choline sufficient; GLDH, glutamate dehydrogenase; GGT, gamma-glutamyl transferase.</p><p>²Pre, main effect of prenatal choline status; Post, main effect of postnatal choline status; Pre x Post, interactive effect of prenatal and postnatal choline statuses.</p><p>Effects of perinatal choline status on clinical blood chemistry profiles of 4-wk-old piglets<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133500#t004fn002" target="_blank">¹</a>.</p

Landmark Variation.

<p>The distance between the atlas and individual brains at the anterior commissure, posterior commissure, and left and right anterior aspect of the caudate. All values are in millimeters.</p><p>Landmark Variation.</p

Location of origin for the atlas. Sagittal, coronal, and axial views of the origin location for the atlas.

<p>Location of origin for the atlas. Sagittal, coronal, and axial views of the origin location for the atlas.</p

Regions of interest.

<p>Sagittal, coronal, and axial slices through the origin showing manually drawn regions of interest.</p