I. Audet

Effect of dietary n-3 fatty acids (fish oils) on boar reproduction and semen quality

The aim of this study was to evaluate the effects of dietary supplementation with different fish oils (rich in PUFA) vs. hydrogenated animal fat (SFA) on semen production and quality, fatty acid composition, and preservation properties in boars under controlled and commercial conditions. In Exp. 1 (in a research station), 44 boars, allocated to 4 dietary treatments, received daily 2.5 kg of basal diet with a supplement of 1) 62 g of hydrogenated animal fat (AF, n = 12); 2) 60 g of menhaden oil containing 18% docosahexanoic acid (DHA) and 15% eicosapentanoic acid (EPA; MO, n = 11); 3) 60 g of tuna oil containing 33% DHA and 6.5% EPA (TO, n = 11); and 4) 60 g of menhaden oil and 2 mg/kg of biotin (MO+B, n = 10). Biotin is a critical factor in the elongation of PUFA. Semen was collected according to 3 successive phases: phase 1 (twice per week for 4 wk); phase 2 (daily collection for 2 wk); and phase 3 (twice per week for 10 wk). Experiment 2 was conducted in commercial conditions; 222 boars were randomly allocated to AF, MO, and TO treatments. Semen was collected twice weekly over a 6-mo period. All diets were balanced to be iso-energetic and provided an equivalent of 989 mg of vitamin E per day. Classical measurements of sperm quantity and quality were done for both experiments. Experiment 1 showed, after 28 wk of supplementation, a massive transfer of n-3 PUFA into sperm from boars fed fish oil diets (MO and TO). No differences were observed among dietary treatments for libido (P > 0.30), sperm production (P > 0.20), or percentage of motile cell (P > 0.20). Unexpectedly, MO+B diet reduced the percentage of normal sperm compared with the other treatments (P < 0.03). In conclusion, although it modified the fatty acid composition of sperm, supplementation of boars with dietary fish oils, rich in long chain n-3 fatty acids, did not influence semen production or quality postejaculation.

Dietary omega-3 fatty acids (fish oils) have limited effects on boar semen stored at 17 °C or cryopreserved

To evaluate the influence of dietary supplementation of omega-3 polyunsaturated fatty acids (n-3 PUFA) on storage of boar semen, three experiments were conducted: two involved long-term, fresh semen storage (Exp. 1 and Exp. 2), whereas the other involved cryopreservation (Exp. 3). Boars were allocated randomly to three dietary treatments (for 6-7 mo). In addition to a daily allowance of 2.5 kg of a basal diet, they received: 1) 62 g of hydrogenated animal fat (AF); 2) 60 g of menhaden oil (MO), containing 18% docosahexanoic acid (DHA) and 15% eicosapentanoic acid (EPA); or 3) 60 g of tuna oil (TO), containing 33% DHA and 6.5% EPA. In Experiment 1 (n = 26) and Experiment 2 (n = 18), semen was cooled and stored in vitro for several days at 17 °C before assessment, whereas in Experiment 3 (n = 18), viability, motility, acrosomal integrity, susceptibility to peroxidation (LPO), and DNA fragmentation were determined in fresh and frozen-thawed sperm. In Experiment 1, sperm from boars fed TO had better resistance to fresh storage; even after 7 or 9 d of storage at 17 °C, there were more (P = 0.03) motile sperm in boars fed TO (>60%) than in those fed AF or MO. In Experiment 2, fish oil supplementation did not influence any aspect of sperm quality during semen storage (P > 0.10). In Experiment 3, cryopreservation decreased the proportion of motile and viable frozen-thawed sperm as well as acrosomal integrity and increased DNA fragmentation and LPO (P < 0.01) relative to fresh semen, although sperm quality was unaffected by treatments (P > 0.09). In conclusion, although adding fish oil to the diet failed to significantly improve the quality of cryopreserved boar sperm, inconsistent responses of long-term storage of cooled sperm to dietary n-3 PUFA supplementation warrant further investigation.

Fish oil diets alter the phospholipid balance, fatty acid composition, and steroid hormone concentrations in testes of adult pigs

The objective was to determine the effect of long-term dietary supplementation of two types of fish oil on lipid composition and steroidogenesis in adult pig testis. Twenty-four Duroc boars, aged 204.5 ± 9.4 d (body weight 128.1 ± 16.7 kg) received daily 2.5 kg of an iso-caloric basal diet supplemented with: 1) 62 g of hydrogenated animal fat (AF); 2) 60 g of menhaden oil (MO) containing 16% of eicosapentaenoic acid (EPA) and 18% of docosahexaenoic acid (DHA); or 3) 60 g of tuna oil (TO) containing 7% of EPA and 33% of DHA. After these diets were consumed for 7 mo, testicular hormones, phospholipid content, and fatty acid composition of individual phospholipids in testis were determined. Body and reproductive organ weights were not significantly affected by dietary treatments. Testicular tissue from boars fed a TO diet, followed by those receiving MO and AF diets, had the lowest level of phosphatidylethanolamine (TO < MO < AF; P < 0.01) but the highest sphingomyelin (TO > MO > AF; P < 0.01). For each phospholipid, boars fed either the MO or TO diet had increased total omega-3 fatty acids, particularly DHA (P < 0.01), by reciprocal replacement of total omega-6 fatty acids (20:4n-6, 22:5n-6). The MO diet increased EPA more than the other diets. Testicular concentrations of testosterone and estradiol were lower in boars fed a TO diet than a MO diet (P < 0.02). In conclusion, long-term dietary supplementation of fish oil, regardless of the EPA/DHA ratio, modified the fatty acid compositions in testis and affected steroid production of healthy adult boars, which may represent a promising models for future studies on fertility.

Effects of dietary vitamin supplementation and semen collection frequency on reproductive performance and semen quality in boars.

The present study was undertaken to assess the relevance of increasing the daily provision of dietary vitamins on vitamin metabolic status and semen characteristics of boars under controlled and commercial conditions as well as to evaluate the efficiency of this vitamin supplement to allow boars to cope with intensive semen collection frequency. In the first experiment, 39 boars were allocated to 2 dietary treatments, a basal diet (control) and the basal diet supplemented with extra fat- and water-soluble vitamins (Vit). Within each treatment, boars were submitted to 2 regimens of semen collection frequency: 3 times per 2 wk (3/2) and 3 times per week (3/1) over a 12-wk period. Afterwards, all boars were intensively collected (daily) for 2 wk. A resting period of 4 wk followed, and all boars were collected 2 times per week. Thereafter, collection frequencies were reversed, and the same procedure was followed until the end of the intensive collection period. A second experiment was conducted in commercial conditions at a commercial stud, and 252 boars were randomly allocated to the control and Vit dietary treatments. All boars were collected 2 times per week over a 6-mo period. Classical measurements of ejaculate and sperm quality were assessed, and blood samples were collected throughout both experiments to quantify vitamin concentrations. In the first experiment, vitamin concentrations in blood and seminal plasma increased in Vit boars (P < 0.05); however, vitamin concentrations were not affected by collection frequency (P > 0.14). The Vit supplement did not affect sperm production or sperm quality (P > 0.28), although semen volume increased during the 12-wk periods for Vit boars (P < 0.05). The 3/1 boars produced fewer doses per ejaculate than 3/2 boars (P < 0.01); however, the cumulative sperm production for the 12-wk periods increased by 19% in 3/1 boars compared with 3/2 boars. In the second experiment, blood plasma concentrations of vitamin B(9) were greater (P < 0.01) in Vit than control boars. The vitamin supplement did not increase sperm production of boars (P > 0.61). In conclusion, dietary supplements of fat- and water-soluble vitamins increase the amount of vitamins available for the animal, and the collection frequencies had no effect on vitamin status. Moreover, in spite of an effect on the ejaculate volume, the dietary supplement of extra vitamins had no effect on sperm production or quality.

Interaction between vitamin B6 and source of selenium on the response of the selenium-dependent glutathione peroxidase system to oxidative stress induced by oestrus in pubertal pig.

This study aimed to assess the interaction between vitamin B6 and selenium (Se) for the flow of Se towards the Se-dependent glutathione peroxidase (GPX) system in response to oxidative stress naturally induced by oestrus in a pubertal pig model. At first oestrus, forty-five gilts were randomly assigned to the experimental diets (n=9/group): basal diet (CONT); CONT+0.3mg/kg of Na-selenite (MSeB60); MSeB60+10mg/kg of HCl-B6 (MSeB610); CONT+0.3mg/kg of Se-enriched yeast (OSeB60); and OSeB60+10mg/kg of HCl-B6 (OSeB610). Blood samples were collected at each oestrus (long-term profiles), and daily from day -4 to +3 (slaughter) of the fourth oestrus (peri-oestrus profiles) after which liver, kidneys, and ovaries were collected. For long-term profiles, CONT had lower blood Se than Se-supplemented gilts (p<0.01) and OSe was higher than MSe (p<0.01). Lower erythrocyte pyridoxal-5-phosphate was found in B60 than B610 (p<0.01). No treatment effect was observed on GPX activity. For peri-oestrus profiles, treatment effects were similar to long-term profiles. Treatment effects on liver Se were similar to those for long-term blood Se profiles and OSe had higher renal Se concentrations than MSe gilts (p<0.01). Gene expressions of GPX1, GPX3, GPX4, and selenocysteine lyase in liver and kidney were greatest in OSeB610 gilts (p<0.05). These results suggest that dietary B6 modulate the metabolic pathway of OSe towards the GPX system during the peri-oestrus period in pubertal pigs.

Fish oil diets do not improve insulin sensitivity and secretion in healthy adult male pigs

The effects of long-term dietary supplementation of fish oil (n-3 PUFA-rich) in adult male pigs on body condition as well as insulin sensitivity and secretion were examined. Fifteen Duroc boars aged 204.5 (sd 9.4) d (body weight 145.8 (sd 16.8) kg) received daily 2.5 kg basal diet with a supplement of: (1) 62 g hydrogenated animal fat (n 5); (2) 60 g menhaden oil containing 10.8 g DHA and 9.0 g EPA (n 6); (3) 60 g tuna oil containing 19.8 g DHA and 3.9 g EPA (n 4). Rations were balanced to be isoenergetic. After 7 months of treatments, oral glucose and meal tolerance tests were conducted after insertion of a catheter into the jugular vein. Dietary supplementation with n-3 PUFA altered the blood plasma profile: DHA and EPA increased whereas arachidonic acid decreased (P < 0.01). Plasma glucose, insulin and C-peptide responses to oral glucose and the test meal were not affected by treatments (P>0.34). For all animals, total body fat estimated from body weight and back fat thickness was correlated with both beta-cell function (by homeostasis model assessment (HOMA); r+0.63) and insulin sensitivity (index of whole-body insulin sensitivity and by HOMA; r - 0.63 and r+0.66, respectively). In conclusion, long-term supplementation with dietary n-3 PUFA did not affect insulin metabolism in healthy adult male pigs. The relationship between body fat and insulin sensitivity, well documented in human subjects, suggests that the adult male pig could be a promising animal model for studies on insulin metabolism.

Effects of dietary vitamin supplementation and semen collection frequency on hormonal profile during ejaculation in the boar.

To evaluate the effect of dietary and management factors on boar hormonal status during ejaculation, 39 boars were canulated to determine the profiles of luteinizing hormone (LH), follicle-stimulating hormone (FSH), 17beta-estradiol (E2), and testosterone (T) in blood plasma and seminal fluid. Prior to canulation, 18 boars were fed a basal diet (control), whereas the remainder (n=21) were fed a basal diet supplemented with extra vitamins (supplemented). Within each dietary treatment, two regimens of semen collection were used over the 3mo preceding the hormonal evaluation: three times per 2wk (3/2) or three times per wk (3/1). Plasma E2 was lower (P<0.01) before ejaculation (232.5+/-22.6pg/mL) than at the onset of ejaculation (255.2+/-27.1ng/mL). Plasma T increased from 5.14+/-0.72, before ejaculation to 5.87+/-0.86ng/mL at the onset of ejaculation in supplemented boars, whereas it decreased from 5.15+/-0.65 to 4.87+/-0.70ng/mL in controls (diet by time, P<0.05). At the onset of ejaculation, plasma FSH was higher in 3/2 boars (0.436+/-0.06ng/mL) than in 3/1 boars (0.266+/-0.04ng/mL; P<0.05). During ejaculation, plasma LH increased linearly (P<0.01) from 0.59+/-0.07 to 0.97+/-0.10ng/mL, and plasma E2 and T concentrations were correlated (r=0.62, P<0.01). Plasma FSH before and during ejaculation was negatively correlated with sperm production (r=-0.60, P<0.01) and testicular weight (r=-0.50, P<0.01). In conclusion, dietary and management factors had few impacts on hormonal profiles during ejaculation, but homeostasis of some hormones was related to some criteria of reproductive performance in boars.

Homocysteine metabolism, growth performance, and immune responses in suckling and weanling piglets

Homocysteine (Hcy), an intermediary sulfur AA, is recognized as a powerful prooxidant with deleterious effects on physiological and immune functions. In piglets, there is an acute 10-fold increase of plasma concentrations of homocysteine (pHcy) during the first 2 wk of life. This project aimed to maximize pHcy variations within physiological ranges using typical supplies of folates and vitamin B12 (B12) to sows and piglets. Growth, immune response, and Hcy metabolism of piglets were studied until piglets reached 56 d of age. Third-parity sows were randomly assigned to a 2 × 2 split-plot design with 2 dietary treatments during gestation and lactation, S(-) (1 mg/kg folates and 20 µg/kg B12, n = 15) and S(+) (10-fold S(-) levels, n = 16), and 2 treatments to piglets within each half litter, intramuscular injections (150 µg) of B12 (P(+)) at d 1 and 21 (weaning) and saline (P(-)). Within each litter of 12 piglets, 3 P(+) and 3 P(-) piglets were studied for growth and Hcy metabolism, and the others were studied for immune responses. During lactation, plasma B12 decreased and was transiently greater in S(+) vs. S(-) piglets on d 1 and P(+) vs. P(-) piglets on d 7 (sow treatment × age and piglet treatment × age; P < 0.05). From 14 to 21 d of age, pHcy was 33% lower in S(+)P(+) vs. S(-)P(-) piglets (sow treatment × piglet treatment interaction; P < 0.05). At 56 d of age, hepatic B12 was greater and pHcy was lower for P(+) vs. P(-) piglets (P < 0.05). No treatment effect was observed on growth except for a lower postweaning G:F in S(+)P(-) piglets than in others (sow treatment × piglet treatment interaction; P < 0.05). Positive correlations were observed between pHcy and growth (r > 0.29, P < 0.05) before and after weaning. Antibody responses to ovalbumin and serum tumor necrosis factor-α were not affected by treatments, but postweaning serum IL-8 peaked earlier in S(-)P(-) vs. S(+)P(+) piglets (piglet treatment × age; sow treatment × piglet treatment interaction, P < 0.05). Proliferation of lymphocytes in response to the mitogen concanavalin A tended to be lower in culture media supplemented with sera from S(-) vs. S(+) piglets (P = 0.081) and P(-) vs. P(+) piglets (P = 0.098), and the reduction of response was more marked (P < 0.05) with high (>21 µM) compared to medium (17 to 21 µM) or low (<17 µM) pHcy. In conclusion, the present vitamin supplements to sows and/or piglets produced variations of pHcy that were not apparently harmful for growth performance of piglets. The greater pHcy, particularly prevalent in S(-) and/or P(-) piglets, had negative effects on some indicators of immune responses, suggesting that these young animals may be immunologically more fragile.

Bioavailability of Vitamin B12 from Dairy Products Using a Pig Model

The present study compares the bioavailability of vitamin B12 (B12) of dairy products or synthetic B12, using the pig as an experimental model for humans. Eleven pigs were used in a cross-over design to assess the net portal drained viscera (PDV) flux of blood plasma B12 after ingestion of tofu (TF; devoid of B12), Swiss cheese (SC), Cheddar cheese (CC), yogurt (YG), and synthetic B12 (TB12; TF supplemented with cyanocobalamin), providing a total of 25 µg of B12 each. PDV blood plasma flow for SC and CC were higher than for TF and TB12 (p ≤ 0.04) whereas YG was higher than TF (p = 0.05). Porto-arterial difference of blood plasma B12 concentrations were higher for CC and TB12 than for TF and YG (p ≤ 0.04) but not different from SC (p ≥ 0.15). Net PDV flux of B12 was only different from zero for CC. However, the net PDV flux of B12 for CC was not different from SC or TB12. Cumulative net PDV flux of B12 for SC, TB12, and CC were 2.9, 4.4, and 8.3 µg 23 h post-meal, corresponding to a bioavailability of 11.6%, 17.5%, and 33.0%, respectively. In conclusion, CC had the best bioavailability of B12 among the tested dairy products or compared to synthetic B12.