Productive and physiological implications of top-dress addition of branched-chain amino acids and arginine on lactating sows and offspring.

BACKGROUND: Branched-chain amino acids (BCAAs), including L-leucine (L-Leu), L-isoleucine (L-Ile), L-valine (L-Val), and L-arginine (L-Arg), play a crucial role in mammary gland development, secretion of milk and regulation of the catabolic state and immune response of lactating sows. Furthermore, it has recently been suggested that free amino acids (AAs) can also act as microbial modulators. This study aimed at evaluating whether the supplementation of lactating sows with BCAAs (9, 4.5 and 9 g/d/sow of L-Val, L-Ile and L-Leu, respectively) and/or L-Arg (22.5 g/d/sow), above the estimated nutritional requirement, could influence the physiological and immunological parameters, microbial profile, colostrum and milk composition and performance of sows and their offspring. RESULTS: At d 41, piglets born from the sows supplemented with the AAs were heavier (P = 0.03). The BCAAs increased glucose and prolactin (P < 0.05) in the sows' serum at d 27, tended to increase immunoglobulin A (IgA) and IgM in the colostrum (P = 0.06), increased the IgA (P = 0.004) in the milk at d 20 and tended to increase lymphocyte% in the sows' blood at d 27 (P = 0.07). Furthermore, the BCAAs tended to reduce the Chao1 and Shannon microbial indices (P < 0.10) in the sows' faeces. The BCAA group was discriminated by Prevotellaceae_UCG-004, Erysipelatoclostridiaceae UCG-004, the Rikenellaceae_RC9_gut_group and Treponema berlinense. Arginine reduced piglet mortality pre- (d 7, d 14) and post-weaning (d 41) (P < 0.05). Furthermore, Arg increased the IgM in the sow serum at d 10 (P = 0.05), glucose and prolactin (P < 0.05) in the sow serum at d 27 and the monocyte percentage in the piglet blood at d 27 (P = 0.025) and their jejunal expression of NFKB2 (P = 0.035) while it reduced the expression of GPX-2 (P = 0.024). The faecal microbiota of the sows in Arg group was discriminated by Bacteroidales. The combination of BCAAs and Arg tended to increase spermine at d 27 (P = 0.099), tended to increase the Igs (IgA and IgG, P < 0.10) at d 20 in the milk, favoured the faecal colonisation of Oscillospiraceae UCG-005 and improved piglet growth. CONCLUSION: Feeding Arg and BCAAs above the estimated requirements for milk production may be a strategy to improve sow productive performance in terms of piglet average daily gain (ADG), immune competence and survivability via modulation of the metabolism, colostrum and milk compositions and intestinal microbiota of the sows. The synergistic effect between these AAs, noticeable by the increase of Igs and spermine in the milk and in the improvement of the performance of the piglets, deserves additional investigation.

Effect of two milk supplements and two ways of administration on growth performance, welfare and fecal microbial ecology of suckling piglets.

Introduction: The aim of this study was to evaluate the effect of two MS formulas, DanMilk™ (AB Neo, Denmark) (MS1) and Neopigg® RescueMilk (Provimi, Netherlands) (MS2) administered manually and to compare two ways of administration (manual vs automatic) of MS1 on growth performance, health, fecal microbial profile, behavior, and skin lesions of piglets during suckling and post-weaning. Methods: Forty litters (528 piglets) were divided into 4 groups: 1) Control group receiving no MS (CON); 2) MS1 administered automatically (A-MS1); 3) MS1 administered manually (Ma-MS1) 4) MS2 administered manually (Ma-MS2). All groups had access to sow milk and creep feed. On day 5 after birth (d0), litters were equalized (13.2 piglets/litter ± 0.8 SD), thereafter no cross-fostering was allowed. Piglets were weighed at day 5 after birth (d0), at the end of milk supplementation (d14), at weaning (d21 of the trial, 26 days of age) and ten days post-weaning (d31). Piglet welfare was assessed using behavioral and lesion measures at d4 and d10. Feces were collected at d14 and d21. Results and discussion: During the suckling period, A-MS1 had lowest mortality (p < 0.05), while Ma-MS1 had lower mortality compared with CON and Ma-MS2 (p < 0.05). Negative social behavior at d4, was more frequent in MS groups (A-MS1, Ma-MS1, Ma-MS2) compared to CON group (p = 0.03). Growth performance and lesion prevalence were not affected by MS provision. During lactation, Ma-MS2 group had a higher percentage of piglets not eating during suckling at d18 compared with Ma-MS1 (p = 0.03). MS1 increased microbial diversity compared with CON at d14 (Chao1, p = 0.02; Shannon, p = 0.03) and compared with CON (Shannon, p < 0.05; InvSimpson, p = 0.01) and Ma-MS2 (Chao1, p < 0.05; Shannon, p = 0.05, InvSimpson p = 0.01) at d21. Groups that received MS1 were characterized by genera producing short-chain fatty acids (SCFAs), i.e., Lachnospiraceae (A-MS1) and Oscillospiraceae (Ma-MS1). MS composition and availability can contribute to reduce piglet's mortality during the suckling phase and can also affect intestinal microbiota by favoring the presence of SCFAs producing bacteria.

Supplementation of mixed doses of glutamate and glutamine can improve the growth and gut health of piglets during the first 2 weeks post-weaning.

The aim of this study was to test the effect of mixing doses of glutamate (Glu) and glutamine (Gln) on the growth, health and gut health of post-weaning piglets. One hundred twenty weaned piglets (24 ± 2 days of age) were assigned to 6 dietary groups: (1) standard diet (CO); (2) CO plus Glu (6 kg/Ton): 100Glu; (3) CO plus 75Glu + 25Gln; (4) CO plus 50Glu + 50Gln; (5) CO plus 25Glu + 75Gln and (6) CO plus 100Gln. At days 8 and 21, blood was collected for haematological and reactive oxygen metabolite analysis, intestinal mucosa for morphological and gene expression analysis, and caecal content for microbial analysis. Data were fitted using a Generalised Linear Model (GLM). Piglet growth increased linearly with an increase in Gln from d7 to d14. The Glu:Gln ratio had a quadratic effect on faecal consistency and days of diarrhoea, neutrophil% and lymphocyte%, and a positive linear effect on monocyte% in the blood at d8. The amino acids (AAs) reduced the intraepithelial lymphocytes in the jejunum, and 100Gln improved intestinal barrier integrity at d8. The caecal microbiota did not differ. Overall, this study suggested a favourable effect of mixing Glu and Gln (25 + 75-50 + 50) as a dietary supplementation in post-weaning piglets to benefit the immune and barrier function of the gut, resulting in an increase in faecal consistency and improvement of growth during the first 2 weeks post-weaning.

Importance of Breed, Parity and Sow Colostrum Components on Litter Performance and Health.

The aims of this study were to investigate the effect of breed and parity on colostrum components, and to associate sow breed, parity, and colostrum components with survival, growth, and the occurrence of diarrhoea of their litters. In Experiment 1, 64 sows (Duroc = 13; Landrace = 17 and Large White = 34) were included. In Experiment 2, 71 sows with different parities (1 = 10; 2 = 16; 3 = 13; 4 = 12; ≥5 = 20) were included. The number (N) of live piglets, litter body weight (Experiment 1), and the occurrence of diarrhoea (Experiment 1) were recorded at farrowing, at 2−3 days of age, and at weaning. Colostrum was analysed for proximate composition, immunoglobulins (Igs), and somatic cell count (SCC). Stepwise regressions and ANOVA models were used to associate breed, parity, and colostrum components with litter performance. The Duroc breed had the highest IgG and IgA (p < 0.005). Gilts had a higher fat% and SCC (p< 0.0001); these compounds were positively correlated (r = 0.45). Increased IgA tended to increase the N of weaned piglets (p = 0.058) and reduce litter diarrhoea (p = 0.021). The SCC increased the N of weaned piglets (p = 0.031). Overall, this study confirmed that breed and parity can influence the colostrum composition and highlighted the key role of Igs and somatic cells in piglet health.

Pattern of Antibiotic Consumption in Two Italian Production Chains Differing by the Endemic Status for Porcine Reproductive and Respiratory Syndrome.

The aim of this case study was to quantify antibiotic (AB) use in Italian weaning (W) and fattening (F) units differentiated for porcine reproductive and respiratory syndrome (PRRS) occurrence. Farms were classified as either PRRS negative (-) or PRRS positive (+) based on the circulation of the virus among the animals. In all the farms, the modified live PRRS virus (PRRSV) vaccine was provided to all the animals. In the PRRS- farms, the level of circulating antibodies was low, and the disease, in its clinical form, did not occur. In the PRRS+ farms, the level of circulating antibodies against the virus was high, and the disease was recurrent. Data regarding AB consumption were collected from 2017 to 2020, and the active compounds (ACs) were expressed as milligrams of AC/total kilogram of body weight (BW) produced. Each AC was classified into one of four categories according to the European Medicines Agency classification of ABs for prudent and responsible use in animals: Avoid, Restrict, Caution, and Prudence. Data regarding the ACs in each category were analyzed using a linear model that included production phase, PRRS status, and their interaction as factors. Performance parameters, average age of the pigs at the end of each phase, daily live weight gain, feed-to-gain ratio, total losses, cost index, and medication costs were significantly influenced by the PRRS chain. The use of class B ABs was not affected by production phase or PRRS status. Conversely, for class C ABs, interaction between the two factors (p = 0.02) was observed; W/PRRS+ and F/PRRS+ showed the greatest AB use for this class (p = 0.003). For class D ABs, the interaction was significant (p = 0.01); class C and D ABs were used more in the weaning (p = 0.07) than in the fattening phase (p = 0.003). For the weaning phase, the use of class C and D ABs was greater in the PRRS+ than in the PRRS- chain (p < 0.01). In conclusion, PRRS status affected the growth of pigs and economic performance. Moreover, PRRS status significantly influenced the use of ABs during all the growing periods with the greatest impact being on the weaning phase.

Weaning differentially affects the maturation of piglet peripheral blood and jejunal Peyer's patches.

The study aimed to assess how the post-weaning condition changes piglet peripheral blood (PB) and jejunal Peyer's patches (JPPs) as compared to the suckling period, and how these changes are associated with intestinal microbiota evolution. Sixteen pigs were slaughtered and sampled for PB, JPPs and jejunal content (JC) at weaning (26 days) or at 12 days fed on a pre-starter diet. The PB and JPP transcriptomes were analysed using mRNA-seq. The Gene Set Enrichment Analysis was used to demonstrate enriched gene clusters, depending on sampling time. Jejunal microbiota was profiled using 16S rRNA gene sequencing. Post-weaning JPPs were enriched for processes related to the activation of IFN-γ and major histocompatibility complex (MHC) class I antigen processing which clustered with the reduced abundance of the Weisella genus and Faecalibacterium prausnitzii in JC. The post-weaning microbiome differed from that seen in just-weaned pigs. For just-weaned PB, the enrichment of genes related to hemoglobin and the iron metabolism indicated the greater presence of reticulocytes and immature erythrocytes. The JPP genes involved in the I MHC and IFN-γ activations were markers of the post-weaning phase. Several genes attributable to reticulocyte and erythrocyte maturation could be interesting for testing the iron nutrition of piglets.

Bacillus spp. Probiotic Strains as a Potential Tool for Limiting the Use of Antibiotics, and Improving the Growth and Health of Pigs and Chickens.

The pressure to increasingly optimize the breeding of livestock monogastric animals resulted in antimicrobials often being misused in an attempt to improve growth performance and counteract diseases in these animals, leading to an increase in the problem of antibiotic resistance. To tackle this problem, the use of probiotics, also known as direct in-feed microbials (DFM), seems to be one of the most promising strategies. Among probiotics, the interest in Bacillus strains has been intensively increased in recent decades in pigs and poultry. The aim of the present review was to evaluate the effectiveness of Bacillus strains as probiotics and as a potential strategy for reducing the misuse of antibiotics in monogastric animals. Thus, the potential modes of action, and the effects on the performance and health of pigs (weaning pigs, lactation and gestation sows) and broilers are discussed. These searches yielded 131 articles (published before January 2021). The present review showed that Bacillus strains could favor growth in terms of the average daily gain (ADG) of post-weaning piglets and broilers, and reduce the incidence of post-weaning diarrhea in pigs by 30% and mortality in broilers by 6-8%. The benefits of Bacillus strains on these parameters showed results comparable to the benefit obtained by the use of antibiotics. Furthermore, the use of Bacillus strains gives promising results in enhancing the local adaptative immune response and in reducing the oxidative stress of broilers. Fewer data were available regarding the effect on sows. Discordant effects have been reported regarding the effect on body weight (BW) and feed intake while a number of studies have supported the hypothesis that feeding probiotics to sows could benefit their reproductive performance, namely the BW and ADG of the litters. Taken all the above-mentioned facts together, this review confirmed the effectiveness of Bacillus strains as probiotics in young pigs and broilers, favoring their health and contributing to a reduction in the misuse of direct in-feed antibiotics. The continuous development and research regarding probiotics will support a decrease in the misuse of antibiotics in livestock production in order to endorse a more sustainable rearing system in the near future.

Investigation of Early Supplementation of Nucleotides on the Intestinal Maturation of Weaned Piglets.

Nucleotides are essential for the development of the gastrointestinal tract and immune function, but their intake with milk by piglets could be insufficient. The effect of nucleotides on growth and health was tested on 98 piglets divided into two groups: NU, orally administrated with 4 mL of a nucleotide-based product (SwineMOD®) at 10, 15, 18, 21, 27 days, or not (CO). Blood and feces were sampled at weaning (26 d, T1), and at 38 d (T2). Per each group and time-point, eight piglets were slaughtered and jejunal Peyer's patches (JPPs) were collected. NU increased hemoglobin content and hematocrit, but not growth. At weaning, the NU fecal microbiota was characterized by the abundance of Campylobacteraceae, more typical of the growing phase, compared to CO, with a greater abundance of Streptococcaceae. For the blood transcriptome, an initial greater inflammatory activation was seen in CO, while at T2, NU enriched gene sets related to erythropoiesis. The activation of gene groups ranging from epigenetic response to transcriptional regulation evidenced an intense proliferative activity in NU JPPs. NU supplementation did not influence the growth performance of piglets but could have expressed a positive effect on pig microbiota anticipating its maturation at weaning. This immunostimulant activity in the JPPs could moderate the inflammation in the immediate pre-weaning.

Functional Amino Acids in Pigs and Chickens: Implication for Gut Health.

In pigs and broiler chickens, the gastrointestinal tract or gut is subjected to many challenges which alter performance, animal health, welfare and livability. Preventive strategies are needed to mitigate the impacts of these challenges on gut health while reducing the need to use antimicrobials. In the first part of the review, we propose a common definition of gut health for pig and chickens relying on four pillars, which correspond to the main functions of the digestive tract: (i) epithelial barrier and digestion, (ii) immune fitness, (iii) microbiota balance and (iv) oxidative stress homeostasis. For each pillar, we describe the most commonly associated indicators. In the second part of the review, we present the potential of functional amino acid supplementation to preserve and improve gut health in piglets and chickens. We highlight that amino acid supplementation strategies, based on their roles as precursors of energy and functional molecules, as signaling molecules and as microbiota modulators can positively contribute to gut health by supporting or restoring its four intertwined pillars. Additional work is still needed in order to determine the effective dose of supplementation and mode of administration that ensure the full benefits of amino acids. For this purpose, synergy between amino acids, effects of amino acid-derived metabolites and differences in the metabolic fate between free and protein-bound amino acids are research topics that need to be furtherly investigated.

The multivariate approach identifies relationships between pre-slaughter factors, body lesions, ham defects and carcass traits in pigs.

Abattoir meat inspection has been proposed for the collection of welfare outcomes. The identification of suitable animal-based measures (ABM) is still a critical point that needs to be implemented to avoid collinearity among measures. The present study aims to benchmark the presence of ABM such as skin and tail lesions and ham defects in carcasses from 79 batches of Italian Heavy pigs and to identify possible relationships between the assessed ABM and pre-slaughter factors such as the season and the overnight lairage. Furthermore, the study also considers the effect of pre-slaughter conditions and ABM on carcass traits parameters (cold carcass weight and lean meat percentage). Skin and tail lesions were recorded at the slaughter line. The presence of abscesses, muscle tears and veining defects were assessed in the hams at trimming, according to the Parma Ham Consortium. A multivariate analysis was performed to identify relationships between ABM and pre-slaughter factor; therefore, a linear model was built to assess the effect on carcass weight and lean meat percentage. Main welfare issues were represented by skin and tail lesions and muscle tears (prevalence above 10%). Multivariate analysis evidenced that skin lesions and veining defects were mostly associated with the warm season. Abscesses and muscle tears in the hams were more likely related to overnight lairage, while tail lesions contributed equally to both season and lairage. Moreover, lairage related factors showed to affect lean meat percentage. The findings of the present study suggest that ham defects might be useful indicators of pre-slaughter stress. The validation of these findings with physiological parameters could be of interest for further studies.

Timely Control of Gastrointestinal Eubiosis: A Strategic Pillar of Pig Health.

The pig gastrointestinal tract (GIT) is an open ecosystem in which microorganisms and their host are mutually involved and continually adapt to different factors and problems which may or may not be host dependent or due to the production system. The aim of the present review is to highlight the factors affecting the GIT microbial balance in young pigs, focusing on the pre- and post-weaning phases, to define a road map for improving pig health and the production efficiency of the food chain. Birth and weaning body weight, physiological maturation, colostrum and milk (composition and intake), genetic background, environmental stressors and management practices, antibiotic use and diet composition are considered. Overall, there is a lack of knowledge regarding the effect that some factors, including weaning age, the use of creep feed, the composition of the colostrum and milk and the use of antibiotics, may have on the gut microbiome of piglets. Furthermore, the information on the gut microbiome of piglets is mainly based on the taxonomy description, while there is a lack of knowledge regarding the functional modification of the microbiota, essential for the exploitation of microbiota potential for modulating pig physiology.

Transfer of Mycotoxins from Lactation Feed to Colostrum of Sows.

Studies regarding the transfer of mycotoxins from sow feed to colostrum are scarce. A sample of in-house produced lactation feed and one of colostrum were collected from two or three sows per farm (total 49) from 19 farms. The feed contents of aflatoxins (AFs), fumonisins (FUs), deoxynivalenol (DON) and zearalenone (ZEA) were assessed using ELISA and confirmed by liquid chromatography-mass spectrometry (LC-MS), The values were very low (10, 12, 17 and 2 positive samples for AFs, FUs, DON and ZEA, respectively), except for two samples (one AF, one DON). Based on feed values, colostrum samples from 13 farms were tested for at least one mycotoxin (Total 35). Aflatoxins were not found in any sample. A signal for FUs was observed in 5 of 11 colostra, despite low feed values; DON was frequently present in the colostrum (10/14). On the farm where the feed exceeded the DON suggested limits, a higher colostrum content was seen, 10.9 µg/kg, approximately 1/69 of the value showing toxicity in young pigs. The absence of reference values for neonate pigs, and the risk of higher and longer ingestion of DON by sows suggested considering routine checks of sow feed; more research on DON transfer and toxicity in piglets is needed.

A Review of the Effect of Formic Acid and Its Salts on the Gastrointestinal Microbiota and Performance of Pigs.

Out of the alternatives to antibiotics and zinc oxide, organic acids, or simply acidifiers, play significant roles, especially in ensuring gut health and the growth performance of pigs. Regarding acidifiers, formic acid and its salts have shown very promising results in weaning, growing and finishing pigs. Although it is known that the main mechanisms by which acidifiers can improve livestock performance and health are related to the regulation of gastrointestinal pH, an improvement in intestinal digestibility and mineral utilization, and their antimicrobial properties against specific pathogens has been observed, while poor consensus remains in relation to the effect of acidifers on bacteria and the complex microbiome. Therefore, the aim of the present review was to critically evaluate the effects of formic acid and its salts on the performance and the gastrointestinal microbiota balance of pigs.

Variations in porcine colostrum oligosaccharide composition between breeds and in association with sow maternal performance.

BACKGROUND: Oligosaccharides (OS) are indigestible carbohydrates naturally found in milk. The composition of porcine colostrum OS may influence the growth and the health of the neonate and consuming optimal concentrations of OS may reduce piglet susceptibility to illness. In this manner, targeted supplementation of animal feed with OS is being explored as a health management tool in the livestock industry. The variation in OS composition between different breeds of pig and its association with the litter performance is currently unknown. The aim of this study was to characterize the colostrum OS composition from sows of different breed and parity and correlate this data with sow maternal traits. METHODS: Eighty-three colostrum samples from parities 1 to 8 were gathered from 3 different breeds of sow: 44 Large White sows, 27 Landrace sows and 12 Duroc sows. Samples were taken between the birth of the first and the last piglet from sows that were not pharmacologically induced to farrow. OS were purified from the samples and analysed by MALDI-ToF mass spectrometry (21 OS compositions detected). The farrowing season and the maternal data were recorded for each sow, including the number of live piglets and the litter body weight at birth, at day (d) 3 and at weaning. RESULTS: Five OS compositions, including isomers of the bifidogenic Sialyllactose, Lacto-N-Tetraose and Lacto-N-Hexaose series, were detected in all the samples. Twelve other OS were identified in at least 50% of samples, and their abundances were affected by breed (P < 0.05; 6 of 12), marginally affected by season (P < 0.10; 3 of 12) and never by parity number. The abundances of each OS component were standardized by Z-score scaling (µ = 0 and SD = 1), transformed by principal component analysis, and four similarity clusters were generated. Cluster membership was associated with litter weight gain within 3 days (P = 0.063) and at weaning (P < 0.05), but not with piglet mortality within 3 days. CONCLUSIONS: OS composition of colostrum may partially explain the variability in maternal performance within and between different breeds of sow. The obtained OS data can provide useful information for the development of novel prebiotic food supplements for suckling and weaning pigs.

Investigation of the Defatted Colostrum 1H-NMR Metabolomics Profile of Gilts and Multiparous Sows and Its Relationship with Litter Performance.

The aim of the study was to characterize the soluble metabolomics profile of defatted colostrum of sows at different parity number (PA) and to correlate the metabolomics profile with the Brix percentage estimate of colostrum immunoglobulin G (IgG) and sow productive traits. A total of 96 Meidam (crossbreed Large White × Meishan) sows of PA from 1-4 (PA1: 28; PA2:26; PA3:12; PA4:26) were included, and their productive traits were recorded at 10 days post-farrowing. Colostrum IgG was quantified using a Brix refractometer, and metabolomics profile was assessed using 1H-NMR spectroscopy. Sows' PA slightly influenced the metabolomics profile of colostrum. lactose and glycine were higher in PA1 compared with PA4 (p 0.05) and N-acetylglucosamine (GlcNAc) tended to be higher in PA2 than PA3 and PA4 (p < 0.10). The Brix percentage of IgG was negatively associated with lactose and positively with creatine, myo-inositol, and O-phosphocholine (p < 0.05). Taurine was positively related to litter weight at birth. GlcNAc and myo-inositol were linked to piglet mortality at day 10 with a negative and positive trend, respectively. In conclusion, colostrum of gilts and multiparous sows had a similar metabolomics profile. Specific metabolites contributed to explanation of the variability in colostrum Brix percentage estimate of IgG concentration and the sows' productive performance.

In ovo Injection of a Galacto-Oligosaccharide Prebiotic in Broiler Chickens Submitted to Heat-Stress: Impact on Transcriptomic Profile and Plasma Immune Parameters.

This study investigated the effects of a galactooligosaccharide (GOS) prebiotic in ovo injected on intestinal transcriptome and plasma immune parameters of broiler chickens kept under thermoneutral (TN) or heat stress (HS) conditions. Fertilized Ross 308 eggs were injected in ovo with 0.2 mL physiological saline without (control, CON) or with 3.5 mg of GOS (GOS). Three-hundred male chicks/injection treatment (25 birds/pen) were kept in TN or HS (30 °C) conditions during the last growing phase, in a 2 × 2 factorial design. At slaughter, from 20 birds/injection group (half from TN and half from HS), jejunum and cecum were collected for transcriptome analysis, and plasma was collected. No differences in plasma parameters (IgA and IgG, serum amyloid) and no interaction between injection treatment and environment condition were found. GOS-enriched gene sets related to energetic metabolism in jejunum, and to lipid metabolism in cecum, were involved in gut barrier maintenance. A homogeneous reaction to heat stress was determined along the gut, which showed downregulation of the genes related to energy and immunity, irrespective of in ovo treatment. GOS efficacy in counteracting heat stress was scarce after ten days of environmental treatment, but the in ovo supplementation modulates group of genes in jejunum and cecum of broiler chickens.

Dose-response of different dietary leucine levels on growth performance and amino acid metabolism in piglets differing for aminoadipate-semialdehyde synthase genotypes.

Dose-response studies of dietary leucine (Leu) in weaners are needed for a proper diet formulation. Dietary Leu effect was assessed in a 3-weeks dose-response trial with a 2 (genotype) x 5 (diets) factorial arrangement on one-hundred weaned pigs (9 to 20 kg body weight (BW)). Pigs differed for a polymorphism at the aminoadipate-semialdehyde synthase (AASS) gene, involved in lysine (Lys) metabolism. Pigs received experimental diets (d7 to d28) differing for the standardized ileal digestible (SID) Leu:Lys: 70%, 85%, 100%, 115%, 130%. Daily feed intake (ADFI), daily gain (ADG) and feed:gain (F:G) in all pigs and ADG and F:G in two classes of BW were analyzed using regression analysis with curvilinear-plateau (CLP) and linear quadratic function (LQ) models. Amino acid (AA) concentrations in plasma, liver, muscle and urine were determined. AASS genotype did not affect the parameters. Dietary Leu affected performance parameters, with a maximum response for ADG and F:G between 100.5% and 110.7% SID Leu:Lys, higher than the usually recommended one, and between 110.5% and 115.4% and between 94.9% and 110.2% SID Leu:Lys for ADG for light and heavy pigs respectively. AA variations in tissues highlighted Leu role in protein synthesis and its influence on the other branched chain AAs.

Bacillus sp. probiotic supplementation diminish the Escherichia coli F4ac infection in susceptible weaned pigs by influencing the intestinal immune response, intestinal microbiota and blood metabolomics.

BACKGROUND: Probiosis is considered a potential strategy to reduce antibiotics use and prevent post-weaning diarrhea (PWD). This study investigated the effect of Bacillus amyloliquefaciens DSM25840 or Bacillus subtilis DSM25841 supplementation on growth, health, immunity, intestinal functionality and microbial profile of post-weaning pigs after enterotoxigenic E. coli (ETEC) F4 challenge. METHODS: Sixty-four post-weaning piglets (7748 g ± 643 g) were randomly allocated to four groups: control basal diet (CO); CO + 1.28 × 106 CFU/g of B. amyloliquefaciens (BAA); CO + 1.28 × 106 CFU/g feed of B. subtilis (BAS); CO + 1 g colistin/kg of feed (AB). At day (d) 7, animals were challenged with 105 CFU/mL of ETEC F4ac O149 and then followed for fecal score and performance until d 21. Blood was collected at d 6, d 12 and d 21 for immunoglobulins, at d 8 for acute phase proteins, at d 8 and d 21 for metabolomics analysis. Jejunum was sampled for morphometry, quantification of apoptosis, cell proliferation, neutral and acid mucine and IgA secretory cells, and microarray analysis at d 21. Jejunum and cecum contents were collected for microbiota at d 21. RESULTS: AB and BAS reduced the fecal score impairment compared to CO (P < 0.05) at d 14. Body weight (BW), average daily weight gain (ADWG), average daily feed intake (ADFI) and gain to feed ratio (G:F) did not differ between Bacillus groups and CO. AB improved BW at d 7, d 14 and d 21, ADWG ADFI and G:F from d 0 to d 7 (P < 0.05). At d 8, CO had higher plasma arginine, lysine, ornithine, glycine, serine and threonine than other groups, and higher haptoglobin than AB (P < 0.05). At d 21, CO had lower blood glycine, glutamine and IgA than BAS. Morphology, cells apoptosis and mucins did not differ. BAS and AB increased the villus mitotic index. Transcriptome profile of BAS and AB were more similar than CO. Gene sets related to adaptive immune response were enriched in BAA, BAS and AB. CO had enriched gene set for nuclear structure and RNA processing. CO had a trend of higher Enterobacteriaceae in cecum than the other groups (P = 0.06). CONCLUSION: Bacillus subtilis DSM25841 treatment may reduce ETEC F4ac infection in weaned piglets, decreasing diarrhea and influencing mucosal transcriptomic profile.

Methodology and application of Escherichia coli F4 and F18 encoding infection models in post-weaning pigs.

The enterotoxigenic Escherichia coli (ETEC) expressing F4 and F18 fimbriae are the two main pathogens associated with post-weaning diarrhea (PWD) in piglets. The growing global concern regarding antimicrobial resistance (AMR) has encouraged research into the development of nutritional and feeding strategies as well as vaccination protocols in order to counteract the PWD due to ETEC. A valid approach to researching effective strategies is to implement piglet in vivo challenge models with ETEC infection. Thus, the proper application and standardization of ETEC F4 and F18 challenge models represent an urgent priority. The current review provides an overview regarding the current piglet ETEC F4 and F18 challenge models; it highlights the key points for setting the challenge protocols and the most important indicators which should be included in research studies to verify the effectiveness of the ETEC challenge. Based on the current review, it is recommended that the setting of the model correctly assesses the choice and preconditioning of pigs, and the timing and dosage of the ETEC inoculation. Furthermore, the evaluation of the ETEC challenge response should include both clinical parameters (such as the occurrence of diarrhea, rectal temperature and bacterial fecal shedding) and biomarkers for the specific expression of ETEC F4/F18 (such as antibody production, specific F4/F18 immunoglobulins (Igs), ETEC F4/F18 fecal enumeration and analysis of the F4/F18 receptors expression in the intestinal brush borders). On the basis of the review, the piglets' response upon F4 or F18 inoculation differed in terms of the timing and intensity of the diarrhea development, on ETEC fecal shedding and in the piglets' immunological antibody response. This information was considered to be relevant to correctly define the experimental protocol, the data recording and the sample collections. Appropriate challenge settings and evaluation of the response parameters will allow future research studies to comply with the replacement, reduction and refinement (3R) approach, and to be able to evaluate the efficiency of a given feeding, nutritional or vaccination intervention in order to combat ETEC infection.

Exploring Differential Transcriptome between Jejunal and Cecal Tissue of Broiler Chickens.

The study proposed an exploratory functional analysis on differential gene expression of the jejunum and of cecum in chickens. For this study, 150 Ross 308 male chickens were randomly allotted in six pens (25 birds/pen) and fed the same commercial diet. From 19 birds of 42 days of age, jejunum and cecum mucosae were collected for RNA extraction for transcriptome microarray analysis. Differentially expressed genes (DEGs) submitted to DAVID (Database for Annotation, Visualization, and Integrated Discovery) and Gene Set Enrichment Analysis (GSEA) software evidenced enriched gene clusters for biological functions differentiated in the tissues. DAVID analysis in the jejunum showed enriched annotations for cell membrane integral components, PPAR (peroxisome proliferator-activated receptor) signaling pathway, and peroxisome and lipid metabolism, and showed DEGs for gluconeogenesis, not previously reported in chicken jejunum. The cecum showed enriched annotations for disulfide bond category, cysteine and methionine metabolism, glycoprotein category, cell cycle, and extracellular matrix (ECM). GSEA analysis in the jejunum showed peroxisome and PPAR signaling pathway-related gene sets, as found with DAVID, and gene sets for immune regulation, tryptophan and histidine metabolism, and renin-angiotensin system, like in mammals. The cecum showed cell cycle and regulation processes, as well as ECM receptor interaction and focal adhesion-related gene sets. Typical intestinal functions specific for the gut site and interesting functional genes groups emerged, revealing tissue-related key aspects which future studies might take advantage of.