Effects of oral supplementation with Spirulina and Chlorella on growth and digestive health in piglets around weaning.

Weaning of piglets is associated with important changes in gut structure and function resulting from stressful events such as separation from the sow, moving to a new facility and dietary transition from a liquid to a solid feed. This may result in post-weaning diarrhoea and a decrease in feed intake and growth. In humans, the cyanobacterium Spirulina platensis (SP) and the freshwater microalga Chlorella vulgaris (CV) are known for their beneficial health effects. This study aimed to determine the effects of early oral administration of Spirulina and Chlorella in piglets on mucosal architecture and cytokine expression in the intestine around weaning, and consequences on growth performance and diarrhoea incidence. The experiment was conducted on 108 suckling piglets of 14 days of age (initial BW=4.9±0.7 kg) and weaned at 28 days of age (day 0). Animals received orally 385 mg/kg BW per day of SP or CV, or water (negative control (NC)) during 4 weeks from day -14 to day 14 and their growth performance was measured daily. After weaning, growth, feed intake and diarrhoea incidence were measured daily. Intestinal morphology and functionality were assessed at day -1, day 2, and day 14. During the suckling period, average daily gain (ADG) in SP piglets was higher, resulting in a higher weaning BW compared to NC and CV piglets (P0.10). Shorter ileal villi were measured in SP and CV piglets than in NC piglets (P<0.05). Cytokine expression did not differ between treatments in response to weaning. At day 14, IL-8 expression in the ileum was higher in SP piglets, while IL-1ß expression in the jejunum was higher in CV piglets (P<0.05). This study shows that Spirulina administration around weaning alleviates diarrhoea in weaned piglets, without marked modulation of local inflammation.

Effects of dietary supplementation with freshwater microalgae on growth performance, nutrient digestibility and gut health in weaned piglets.

In pigs, digestive disorders associated with weaning lead to antibiotic use to maintain intestinal health. Microalgae have been studied in humans and rodents for their beneficial effects on health. The nutritional value of microalgae in animal diets has been assessed, but results were not conclusive. Dietary supplementation with microalgae as an alternative to antibiotic use was studied in two trials (72 piglets with initial BW=9.1±1.1 kg in trial 1 and 24 piglets with initial BW=9.1±0.9 kg in trial 2). All piglets were weaned at 28 days of age and then housed in individual cages. Piglets were randomly allocated to one of the four diets during 2 weeks after weaning: a standard diet with no supplementation (NC) or the standard diet supplemented with 1% Spirulina (SP), with 1% Chlorella (CV), or with 0.2% of colistin as positive control (PC). Trial 1 was performed to determine the effect of microalgae supplementation from 28 to 42 days on performance and incidence of diarrhoea. Animals received then a standard diet from 42 to 56 days of age. Trial 2 was performed from 28 to 42 days of age to assess nutrient digestibility of the experimental diets and to determine inflammatory status and intestinal morphology at 42 days of age. In trial 1, 94% of the pigs had diarrhoea in the 1st week after weaning with no beneficial effect of colistin on diarrhoea incidence, average daily feed intake (ADFI), average daily gain (ADG), and gain : feed (G : F) ratio. This suggests that the diarrhoea was due to digestive disorders that did not result from enterotoxigenic Escherichia coli infection. Supplementation with either Spirulina or Chlorella did not affect ADFI, ADG and G : F in trials 1 and 2 (P>0.10). Diarrhoea incidence was reduced in CV pigs compared with NC, SP and PC pigs (P<0.05). Total tract digestibility in pig receiving microalgae was greater for gross energy (P<0.05), and tended to be greater for dry matter, organic matter and NDF (P<0.10) compared with NC and PC pigs. Villus height at the jejunum was greater in SP and CV pigs compared with NC and PC pigs (P<0.05). This study shows a potential effect of both Spirulina and Chlorella supplementation on intestinal development and a potential of Chlorella supplementation to manage mild digestive disorders. Further investigation is necessary to determine the mechanism action of Spirulina and Chlorella on gut health and physiology.

Performance of piglets in response to the standardized ileal digestible phenylalanine and tyrosine supply in low-protein diets.

Reducing the CP level of the diet allows for a reduction in N excretion without limiting performance as long as the amino acid (AA) requirements are covered. The availability of crystalline AA has permitted for a considerable reduction in the CP level of diets, practically used in pig nutrition. The adoption of low CP diets and the extent to which the CP content can be reduced further depends on the knowledge of the minimum levels of indispensable AA that maximize growth. The standardized ileal digestible (SID) Phe : Lys and Tyr : Lys requirements and the possibility to substitute Tyr by Phe have never been studied in piglets. The objectives of this study were to estimate these requirements in 10 to 20 kg pig as well as to determine the extent to which Phe can be used to cover the Tyr requirement. In three dose-response studies, six pigs within each of 14 blocks were assigned to six low CP diets (14.5% CP) sub-limiting in Lys at 1.00% SID. In experiment 1, the SID Phe : Lys requirement estimate was assessed by supplementing a Phe-deficient diet with different levels of l-Phe to attain 33%, 39%, 46%, 52%, 58%, and 65% SID Phe : Lys. Because Phe can be used for Tyr synthesis, the diets provided a sufficient Tyr supply. A similar approach was used in experiment 2 with six levels of l-Tyr supplementation to attain 21%, 27%, 33%, 39%, 45% and 52% SID Tyr : Lys. Phenylalanine was supplied at a level sufficient to sustain maximum growth (estimated in experiment 1). The SID Phe : Lys and SID Tyr : Lys requirements for maximizing daily gain were 54% and 40% using a curvilinear-plateau model, respectively. A 10% deficiency in Phe and Tyr reduced daily gain by 3.0% and 0.7%, respectively. In experiment 3, the effect of the equimolar substitution of dietary SID Tyr by Phe to obtain 50%, 57%, and 64% SID Phe : (Phe+Tyr) was studied at two limiting levels of Phe+Tyr. From 57% to 64% SID Phe : (Phe+Tyr), performance was slightly reduced. In conclusion, it is recommended not to use a Phe+Tyr requirement in the ideal AA profile but rather use a SID Phe : Lys of 54% and a SID Tyr : Lys of 40% to support maximal growth.

The use of free amino acids allows formulating very low crude protein diets for piglets.

Reducing the dietary CP level with free AA supplementation allows reduction of N excretion and the risk of gut disorders while maintaining performance of pigs. We have recently reevaluated the Val, Ile, His, and Leu requirements in pigs, which enables formulating very low CP diets. However, requirements for undifferentiated N, and thus the extent to which the dietary CP content can be reduced, are unknown. Two experiments were conducted to test the effect of reducing the dietary CP content to meet an ideal AA profile on N retention and performance in 10- to 20-kg pigs. In the first experiment, we measured N retention in 6 blocks of 4 pigs each, all receiving diets with 1.15% standardized ileal digestible (SID) Lys. Each pig within a block was allotted to different levels of dietary CP (19.7%, 16.8%, 14.0%, and 12.7%). The reduction of CP content from 19.7% to 16.8% had no impact on N retention and decreased N excretion by 29%. A further decrease in CP content to 14.0% and 12.7% resulted in a reduced N retention (P < 0.01). In the second experiment, we measured performance in 14 groups of 6 pigs each, allotted to 6 levels of dietary CP with 1.00% SID Lys with soybean meal (17.6%, 15.6%, 13.5%, and 11.8%) or without soybean meal (14.0% and 13.0%). Decreasing the dietary CP content from 17.6% to 13.5% had no effect on performance. Daily gain was reduced in pigs receiving the 11.8% CP diet (P < 0.01). Feeding the diet formulated without soybean meal with 13.0% CP content resulted in reduced feed efficiency, but the addition of Glu to increase the CP content from 13.0% to 14.0% restored performance (P < 0.01). In conclusion, the use of L-Val, L-Leu, L-Ile, L-His, and L-Phe enables a 4% unit reduction in the CP content relative to SID Lys in diets for pigs. Soybean meal can be totally replaced using cereals and free AA. However, a further reduction in dietary CP:Lys level below 13.5% reduces feed efficiency, indicating that the supply of N for the synthesis of dispensable AA may be a limiting factor for the performance of pigs.

Meal patterns in relation to the supply of branched-chain amino acids in pigs.

When a diet deficient in Val with excessive Leu is offered to pigs, feed intake is reduced. The physiological mechanisms underlying the rejection of branched-chain AA (BCAA) unbalance are unknown. A meal pattern analysis was performed to determine how pigs respond to the ingestion of a diet unbalanced in BCAA to identify the mechanisms responsible for the aversive response of such a diet. Eight female pigs (initial BW of 30.7±1.7 kg at 9 wk of age) received alternatively a diet either unbalanced in BCAA supply (i.e., a diet deficient in Val with an excess supply of Leu) or a control diet (i.e., a diet with a sufficient supply of Val with an excess supply of Leu) during 4 periods of 5 consecutive days. Feed consumption was monitored continuously to determine feeding behavior. Average daily feed intake of pigs receiving the diet unbalanced in BCAA was l3% less than that in pigs offered the control diet (P<0.01). Offering the unbalanced diet had no effect on meal size but reduced the number of meals from 9.9 to 8.5 per day (P<0.01). There was no difference between diets in intermeal interval but a slight trend for an increased meal duration from 14.5 to 16.3 min in pigs receiving the unbalanced diet (P=0.13). The rejection of feed appeared within the first 2 d after offering the unbalanced diet. The results of this experiment indicate that the BCAA unbalanced diet had no effect on satiety and satiation. After initiation of a meal, pigs ate a normal-sized meal and were hungry, but the increase in meal duration and a decrease in the number of meals inferred a negative alliesthesia (less pleasant) response to the BCAA unbalanced diet. On the basis of meal patterns, the rejection of a diet unbalanced in BCAA may not involve short-term mechanisms related to oropharyngeal and gastrointestinal receptors but may be triggered by postabsorptive signals.

Response of piglets to the standardized ileal digestible isoleucine, histidine and leucine supply in cereal-soybean meal-based diets.

Improving the amino acid (AA) profile of the diet by using l-Lys, l-Thr, dl-Met, l-Trp and l-Val helps to reduce the dietary CP content, thereby reducing nitrogen excretion while maintaining the performance of pigs. Valine is the fifth limiting AA in cereal-soybean meal-based diets. The extent to which the CP content in the diet can be reduced further without compromising performance depends on the requirement of the next limiting AA. In cereal-soybean meal-based diets, Ile, His and Leu may be the limiting AAs after Val, although information on the requirements for these AAs is scarce. Six experiments were conducted to determine the effect of supplementing a low-CP diet with l-Ile, l-His and l-Leu on the performance of pigs weighing 10 to 20 kg. Experiment 1 was designed to determine the most limiting AA with respect to performance among Ile, His and Leu. A diet 10% deficient in Ile, Leu and His relative to the National Research Council (NRC, 1998) requirement estimates tended to decrease daily feed intake and daily gain by 6% and 8%, respectively. A 10% deficiency in His alone had no effect, whereas a 10% deficiency in Ile or Leu slightly reduced daily feed intake and gain. In the remaining experiments, the standardized ileal digestible (SID) Ile : Lys, His : Lys and Leu : Lys requirements were estimated. In Experiments 2, 3, 4, 5 and 6, 14 blocks of six pigs each were assigned to six levels of SID Ile : Lys (40%, 43%, 46%, 49%, 52% and 55%), His : Lys (20%, 24%, 28%, 32%, 36% and 40%), His : Lys (21%, 24%, 27%, 30%, 33% and 36%), Leu : Lys (70%, 78%, 86%, 94%, 102% and 110%) and Leu : Lys (80%, 90%, 100%, 110%, 120% and 130%), respectively. Across experiments, the estimated SID Ile : Lys, His : Lys and Leu : Lys requirements for maximizing daily gain were 49%, 32% and 102%, respectively, using a curvilinear plateau model. When Ile, His and Leu levels were 10% below the requirement estimate, daily gain was reduced by 9%, 3% and 3%, respectively. The results of this study indicate that the Ile requirement estimate is lower than the current NRC requirement estimate, whereas the Leu and His requirements correspond to those proposed by the NRC.

Meta-analysis of the response of growing pigs to the isoleucine concentration in the diet.

The efficiency of nitrogen utilization will be highest when the amino acid (AA) supply approaches the requirement of the animal. With the availability of different crystalline AA, it is theoretically possible to formulate low-protein diets for growing pigs in which seven AA are co-limiting for performance. In such a diet, the concentration of Lys, Met, Met + Cys, Thr, Trp and Val and a seventh AA would exactly match the requirement. To determine the extent to which low-protein diets can be used, it is important to have reliable information about the requirements for these AA. Isoleucine is often considered the seventh-limiting AA in diets for growing pigs; however, information about the Ile requirement is limited and sometimes conflicting. The purpose of this study was to carry out a meta-analysis of the available literature information to determine the Ile requirement in growing pigs. A total of 46 Ile dose-response experiments were identified that used at least four concentrations of Ile in the diet. Because of differences in experimental design, both the Ile concentration and the response criteria were standardized. In 13 dose-response experiments, there was no indication of a response to an increasing Ile concentration. For the other 33 experiments, a response to the increasing Ile concentration was observed and the Ile requirement estimates ranged from 53% to 114% of that of the National Research Council (1998). An Ile concentration below the requirement resulted in important reductions in both feed intake and growth. A 10% reduction in the Ile concentration (below the requirement) resulted in a 15% reduction in feed intake and a 21% reduction in daily gain. The use of blood products in the diet was the main factor determining whether a response to the Ile concentration was observed or not. Blood meal and blood cells are protein sources with a very low Ile concentration, but with high or very high concentrations of Leu, Val, Phe and His. Some of these AA compete with Ile for catabolic pathways or transport across the blood-brain barrier, thereby potentially increasing the requirement for Ile. In diets without blood products, the Ile requirement appears to be lower than the currently recommended requirement. On the basis of the outcome of this study, we recommend a Ile : Lys requirement ratio of at least 50% on a standardized ileal digestible basis.

Providing a diet deficient in valine but with excess leucine results in a rapid decrease in feed intake and modifies the postprandial plasma amino acid and α-keto acid concentrations in pigs.

Indispensable AA are involved in the control of feed intake. When a diet deficient in Val is offered to pigs, feed intake is typically reduced. This effect is aggravated when dietary Leu is supplied in excess of the requirement. If an unbalanced supply of branched-chain AA (BCAA) is harmful, an anorectic response may serve as a mechanism to prevent this situation. We verified this hypothesis by measuring the voluntary feed intake of a balanced diet offered during the 30-min period 1 h after ingestion of a test meal deficient or not in Val (Val- and Val+) with an excess of Leu. Twelve and four 6-wk-old crossbred female pigs were used in Exp. 1 and 2, respectively. Prior ingestion of the Val- test meal resulted in a 14% reduction in feed intake compared with that observed after ingestion of the Val+ test meal (P = 0.06) in Exp. 1, indicating that the signal to reduce feed intake occurred within 1 h. It is possible that the plasma concentration of the limiting AA serves as a signal for the dietary AA deficiency. We therefore determined the postprandial plasma concentrations of BCAA and their α-keto acids after ingestion of Val- and Val+ in 4 pigs in Exp. 2. After ingestion of the Val- diet, plasma concentrations of Val and its keto acid were reduced compared with values observed after ingestion of the Val+ diet. The peak concentration occurred earlier after ingestion of the Val- diet compared with that of the Val+ diet. Although the plasma concentration increased after the meal, it declined rapidly in pigs offered Val-, and the Val concentration 4 h after ingestion of the meal was even less than that observed in the fasted state. In conclusion, it appears that the pig is able to detect a deficient supply of Val within 1 h after ingestion. The plasma concentration of Val or its concentration relative to the other BCAA during the postprandial period may act as a signal indicating the AA deficiency.

Linoleic and α-linolenic acid as precursor and inhibitor for the synthesis of long-chain polyunsaturated fatty acids in liver and brain of growing pigs.

Studies suggested that in human adults, linoleic acid (LA) inhibits the biosynthesis of n-3 long-chain polyunsaturated fatty acids (LC-PUFA), but their effects in growing subjects are largely unknown. We used growing pigs as a model to investigate whether high LA intake affects the conversion of n-3 LC-PUFA by determining fatty acid composition and mRNA levels of Δ5- and Δ6 desaturase and elongase 2 and -5 in liver and brain. In a 2 × 2 factorial arrangement, 32 gilts from eight litters were assigned to one of the four dietary treatments, varying in LA and α-linolenic acid (ALA) intakes. Low ALA and LA intakes were 0.15 and 1.31, and high ALA and LA intakes were 1.48 and 2.65 g/kg BW0.75 per day, respectively. LA intake increased arachidonic acid (ARA) in liver. ALA intake increased eicosapentaenoic acid (EPA) concentrations, but decreased docosahexaenoic acid (DHA) (all P < 0.01) in liver. Competition between the n-3 and n-6 LC-PUFA biosynthetic pathways was evidenced by reductions of ARA (>40%) at high ALA intakes. Concentration of EPA (>35%) and DHA (>20%) was decreased by high LA intake (all P < 0.001). Liver mRNA levels of Δ5- and Δ6 desaturase were increased by LA, and that of elongase 2 by both ALA and LA intakes. In contrast, brain DHA was virtually unaffected by dietary LA and ALA. Generally, dietary LA inhibited the biosynthesis of n-3 LC-PUFA in liver. ALA strongly affects the conversion of both hepatic n-3 and n-6 LC-PUFA. DHA levels in brain were irresponsive to these diets. Apart from Δ6 desaturase, elongase 2 may be a rate-limiting enzyme in the formation of DHA.

Response of piglets to the valine content in diet in combination with the supply of other branched-chain amino acids.

The branched-chain amino acids (BCAA) valine (Val) and isoleucine (Ile) are considered to be among the next-limiting amino acids for growth in piglets. In earlier studies, we estimated the standardized ileal digestible (SID) Val : Lys (lysine) requirement to be at least 70%, whereas the Ile : Lys requirement may be as low as 50%. Because the BCAA partially share a common route of catabolism, the supply of one BCAA may affect the availability of the other BCAA. Four experiments were conducted to determine the response of 6-week-old piglets to the Val supply in relation to the other BCAA. A deficient supply of Val or Ile typically results in a reduction in average daily feed intake (ADFI). Experiment 1 was designed to determine the effect of a limiting Val supply, independent of the effect on feed intake. In a dose-response study using restrictively fed piglets, nitrogen retention did not increase for an SID Val : Lys supply greater than 64%. In the remaining experiments, piglets were offered feed ad libitum using ADFI, average daily gain (ADG) and gain-to-feed ratio as response criteria. The interaction between the Val and leucine (Leu) was studied in Experiment 2 in a 2 × 2 factorial design (60% and 70% SID Val : Lys, and 111% and 165% SID Leu : Lys). Performance was considerably lower in piglets receiving 60% Val : Lys compared with those receiving 70% Val : Lys and was lowest in piglets receiving the diet with low Val and high Leu content. To further evaluate the interaction between Val and Leu, a dose-response study was carried out in which the response to Val supply was studied in combination with high Leu supply (165% Leu : Lys). Using a curvilinear-plateau model, the average SID Val : Lys requirement was 72%. However, low Val supply (60% SID Val : Lys) reduced performance by 13% to 38%, which was much greater than what we observed in earlier studies. Experiment 4 was carried out to test the hypothesis that the Val requirement is not affected by low Ile supply (50% SID Ile : Lys). Performance was not improved for Val : Lys supplies greater than 65%, which may indicate that Ile (and not Lys) was second-limiting in this study. In conclusion, the first response of piglets to deficient Val supply appears to be a reduction in ADFI, rather than a reduction in ADG or nitrogen retention. A large supply of Leu may not affect the Val requirement per se, but may aggravate the consequences of Val deficiency.