Search

The effect of long-term administration of two Bacillus strains was tested on 98 breeding sows and their litters allotted into three treatments: a control group (CON); supplemented with 5 × 108 cfu/kg B. subtilis - 541 (BSU); or with 5 × 108 cfu/kg B. amyloliquefaciens - 516 (BAM). Reproductive and performance variables were recorded over three cycles with 56 dams remaining through the third lactation. Blood and fecal samples were taken longitudinally from 12 sows per treatment on days 8 and 21 of the third lactation and milk samples were taken on day 21. Feces from one piglet per litter was sampled on days 21 and 33 and jejunal gene expression was assessed in two piglets on day 21. Changes in fecal microbiota were assessed by 16S rRNA gene sequencing (Illumina MiSeq) and gene expression by Open-Array technology. Metabolomic responses were analyzed in milk by NMR and Ig-G and Ig-A specific antibodies were determined by ELISA. No significant differences were observed on feed intake, body weight, or fat mobilization of the sows. However, a significant increase in the total number of piglets born was observed in supplemented sows. Although the increase was seen from the first cycle with BAM, improvements were not seen with BSU until the third cycle. BAM also increased the number of born-alive and weaned piglets. NMR analysis showed an impact of BAM on milk composition. No differences were found in milk or blood immunoglobulins. A different structure of the fecal microbiota was found in supplemented sows, with changes across phylum, family, and genus. These changes were greater at day 8, suggesting a relevant role of probiotics establishing a new intestinal balance after labor. Shifts in the microbiota were also seen in the piglets, with a clearer impact post-weaning than in suckling. In this regard, correlations between microbial groups of sows and piglets showed a higher link with weaned (d33) than with suckling pigs (d21), reinforcing the idea of an early maternal carry-over. No changes due to treatment in jejunal gene expression were detected; however, piglet size had a clear impact on different genes. In summary, the addition of both probiotics, and particularly Bacillus amyloliquefaciens, demonstrated potential benefits on the prolificacy of sows. Daily feeding of Bacillus amyloliquefaciens resulted in an increase in the number of weaned piglets. The high correlations between the compositions of the microbiota of sows and their piglets are evidence of maternal imprinting, with effects lasting beyond weaning., This research was co-funded by the Ministry of Economy, Industry, and Competitiveness (MINECO) of Spain (project code AGL2016-75463-R) within the framework of Proyectos I+D+I Convocatoria RETOS 2016 and the State Plan for Scientific and Technical Research and Innovation. M.S. received an FPI grant from the Spanish Ministry of Science and Innovation (grant number BES-2017-080018). M.D. received support from Opening Sphere UAB-CEI to Postdoctoral Fellows (project H2020-MSCA-COFUND-2014). M.F. received financial support from the Ministry of Agriculture of the Czech Republic (QK1810463). D.S.O. received financial support from the UAB-Banco de Santander Talent Program. We also thank Chr.Hansen A/S for providing the material (probiotics) to carry out this trial.

Short and medium-chain fatty acids (SCFA and MCFA, respectively) are commonly used as feed additives in piglets to promote health and prevent post-weaning diarrhoea. Considering that the mechanism ...

The use of bifidobacteria as probiotics has proven to be beneficial in gastroenteric infections. Furthermore, prebiotics such as inulin can enhance the survival and growth of these bacteria. Two trials were performed to evaluate the effects of the administration of

Background The search for alternatives to antibiotics in pig production has increased the interest in natural resources with antimicrobial properties, such as medium-chain fatty acids (MCFA) as in-feed additives. This study evaluated the potential of a novel blend of MCFA salts (DIC) from distilled coconut oil with a lauric acid content to reduce enteropathogens and control intestinal diseases around weaning. Two experimental disease models were implemented in early-weaned piglets, consisting of two oral challenges: Salmonella Typhimurium (1.2 × 108 CFU) or enterotoxigenic Escherichia coli (ETEC) F4 (1.5 × 109 CFU). The parameters assessed were: animal performance, clinical signs, pathogen excretion, intestinal fermentation, immune-inflammatory response, and intestinal morphology. Results The Salmonella challenge promoted an acute course of diarrhea, with most of the parameters responding to the challenge, whereas the ETEC F4 challenge promoted a mild clinical course. A consistent antipathogenic effect of DIC was observed in both trials in the hindgut, with reductions in Salmonella spp. plate counts in the cecum (P = 0.03) on d 8 post-inoculation (PI) (Salmonella trial), and of enterobacteria and total coliform counts in the ileum and colon (P < 0.10) on d 8 PI (ETEC F4 trial). When analyzing the entire colonic microbiota (16S rRNA gene sequencing), this additive tended (P = 0.13) to reduce the Firmicutes/Bacteroidetes ratio and enriched Fibrobacteres after the Salmonella challenge. In the ETEC F4 challenge, DIC prompted structural changes in the ecosystem with increases in Dialister, and a trend (P = 0.14) to increase the Veillonellaceae family. Other parameters such as the intestinal fermentation products or serum pro-inflammatory mediators were not modified by DIC supplementation, nor were the histological parameters. Only the intraepithelial lymphocyte (IEL) counts were lowered by DIC in animals challenged with Salmonella (P = 0.07). With ETEC F4, the IEL counts were higher with DIC on d 8 PI (P = 0.08). Conclusions This study confirms the potential activity of this MCFA salts mixture to reduce intestinal colonization by opportunistic pathogens such as Salmonella or E. coli and its ability to modulate colonic microbiota. These changes could explain to some extent the local immune cell response at the ileal level.

Probiotics are applied in all the different productive phases of swine production. They are used for multiple purposes, such as improving performance, mitigating diseases, increasing quality of the product or reducing environmental pollutants. Still, their use is not a general tendency in the pig industry. Probiotics are skeptically seen by many nutritionists due to their highly variable results and unpredictable outcomes, when they are used empirically. Recent technological developments, however, have enabled considerable progress in probiotic research in the last few decades. Significant advances have been made in the selection and characterization of specific probiotic cultures, together with the substantiation of claims relating to their use. It therefore seems opportune to conduct a general review concerning the potential role of probiotics in current pig production, analyzing the dimension of their proven effects and providing scientific background to their use. Furthermore, an analysis of strengths, weaknesses, opportunities and threats has been performed in order to assess the prospective role of probiotics in the near future. Many factors can be seen in favor of probiotics, including the capacity to modulate health and improve performance, important technological development and multiple possibilities to enhance their effects. Moreover, they are considered “natural” alternatives to antibiotics and, therefore, are positively supported by customer and legislative pressure. Nevertheless, aspects such as the lack of universal effects, scarce viability in farm conditions, not fully guaranteed safety and exigent regulations have been outlined as the main weak points of probiotics to become fully established in swine production.

The aim of this study was to determine the possible impact of early socialization and an enriched neonatal environment to improve adaptation of piglets to weaning. We hypothesized that changes in the microbiota colonization process and in their metabolic response and intestinal functionality could help the animals face weaning stress. A total of 48 sows and their litters were allotted into a control (CTR) or an enriched treatment (ENR), in which piglets from two adjacent pens were combined and enriched with toys. The pattern of caecal microbial colonization, the jejunal gene expression, the serum metabolome and the intestinal physiology of the piglets were assessed before (-2 d) and after weaning (+ 3d). A differential ordination of caecal microbiota was observed after weaning. Serum metabolome suggested a reduced energetic metabolism in ENR animals, as evidenced by shifts in triglycerides and fatty acids, VLDL/LDL and creatine regions. The TLR2 gene showed to be downregulated in the jejunum of ENR pigs after weaning. The integration of gene expression, metabolome and microbiota datasets confirmed that differences between barren and enriched neonatal environments were evident only after weaning. Our results suggest that improvements in adaptation to weaning could be mediated by a better response to the post-weaning stress., We would like to acknowledge the funding from the Ministry of Economy, Industry and Competitiveness (MINECO) of Spain for granting the project AGL2016-75463-R within the framework of Proyectos I+D+I Convocatoria RETOS 2016 and the State Plan for Scientific and Technical Research and Innovation. MS received an FPI grant from the Spanish Ministry of Science and Innovation (grant number BES-2017-080018). MD received support from Opening Sphere UAB-CEI to Postdoctoral Fellows (project H2020-MSCA-COFUND-2014). HLK received the University, Research Center and Hospital Foundation Grants for the Contracting of New Research Staff (FI) from Agency for Management of University and Research Grants (AGAUR) of the Catalan Government. PL received support from the Tecniospring program (TECSPR15-1-0040) of ACCIÓ funded by the Catalan Government and the Marie Curie COFUND Fellowship Program within the 7th European Community Framework.

The inhibition of the attachment of bacteria to the intestine by receptor analogues could be a novel approach to prevent enterotoxigenic Escherichia coli (ETEC) K88-induced diarrhoea in piglets. The objective of the present study was to screen the ability of different feed ingredients (FI) to bind to ETEC K88 (adhesion test, AT) and to block its attachment to the porcine intestinal mucus (blocking test, BT) using in vitro microtitration-based models. In the AT, wheat bran (WB), casein glycomacropeptide (CGMP) and exopolysaccharides exhibited the highest adhesion to ETEC K88 (P, The present study was supported by the public research project of the Spanish Ministry of Education and Science (Project AGL 2009-07328). The Spanish Ministry of Education and Science had no role in the design and analysis of the study or in the writing of this article.

We evaluated the potential of multi-strain probiotic (Bifidobacterium longum subsp. infantis CECT 7210 and Lactobacillus rhamnosus HN001) with or without galacto-oligosaccharides against enterotoxigenic Escherichia coli (ETEC) F4 infection in post-weaning pigs. Ninety-six piglets were distributed into 32 pens assigned to five treatments: one non-challenged (CTR+) and four challenged: control diet (CTR−), with probiotics (>3 × 1010 CFU/kg body weight each, PRO), prebiotic (5%, PRE), or their combination (SYN). After 1 week, animals were orally inoculated with ETEC F4. Feed intake, weight, and clinical signs were recorded. On days 4 and 8 post-inoculation (PI), one animal per pen was euthanized and samples from blood, digesta, and tissues collected. Microbiological counts, ETEC F4 real-time PCR (qPCR) quantification, fermentation products, serum biomarkers, ileal histomorphometry, and genotype for mucin 4 (MUC4) polymorphism were determined. Animals in the PRO group had similar enterobacteria and coliform numbers to the CTR+ group, and the ETEC F4 prevalence, the number of mitotic cells at day 4 PI, and villus height at day 8 PI were between that observed in the CTR+ and CTR− groups. The PRO group exhibited reduced pig major acute-phase protein (Pig-MAP) levels on day 4 PI. The PRE diet group presented similar reductions in ETEC F4 and Pig-MAP, but there was no effect on microbial groups. The SYN group showed reduced fecal enterobacteria and coliform counts after the adaptation week but, after the inoculation, the SYN group showed lower performance and more animals with high ETEC F4 counts at day 8 PI. SYN treatment modified the colonic fermentation differently depending on the MUC4 polymorphism. These results confirm the potential of the probiotic strains and the prebiotic to fight ETEC F4, but do not show any synergy when administered together, at least in this animal model.

Simple Summary Weaning is an important period for the swine industry and is influenced by the early events that occur during gestation and lactation. Therefore, a range of dietary and management strategies have to be implemented to achieve optimal health status, maturity, and weight at weaning. In this review, we aimed to identify the major dietary nutrients and management strategies to enhance fetal growth, reducing the oxidative and inflammatory status of sows, modulating the microbiota of sows, enhancing colostrum and milk production, and taking care of neonatal piglets. Abstract The performance of piglets in nurseries may vary depending on body weight, age at weaning, management, and pathogenic load in the pig facilities. The early events in a pig’s life are very important and may have long lasting consequences, since growth lag involves a significant cost to the system due to reduced market weights and increased barn occupancy. The present review evidences that there are several strategies that can be used to improve the performance and welfare of pigs at weaning. A complex set of early management and dietary strategies have been explored in sows and suckling piglets for achieving optimum and efficient growth of piglets after weaning. The management strategies studied to improve development and animal welfare include: (1) improving sow housing during gestation, (2) reducing pain during farrowing, (3) facilitating an early and sufficient colostrum intake, (4) promoting an early social interaction between litters, and (5) providing complementary feed during lactation. Dietary strategies for sows and suckling piglets aim to: (1) enhance fetal growth (arginine, folate, betaine, vitamin B12, carnitine, chromium, and zinc), (2) increase colostrum and milk production (DL-methionine, DL-2-hydroxy-4-methylthiobutanoic acid, arginine, L-carnitine, tryptophan, valine, vitamin E, and phytogenic actives), (3) modulate sows’ oxidative and inflammation status (polyunsaturated fatty acids, vitamin E, selenium, phytogenic actives, and spray dried plasma), (4) allow early microbial colonization (probiotics), or (5) supply conditionally essential nutrients (nucleotides, glutamate, glutamine, threonine, and tryptophan).

This study evaluates the efficacy of two plant-based feed supplementations to fight colibacillosis in weanlings. A total of 96 piglets (32 pens) were assigned to four diets: a control diet (T1) or supplemented with ZnO (2500 ppm Zn) (T2) or two different plant supplements, T3 (1 kg/t, based on essential oils) and T4 (T3 + 1.5 kg/t based on non-volatile compounds). After one week, animals were challenged with ETEC F4, and 8 days after, one animal per pen was euthanized. Performance, clinical signs, microbial analysis, inflammatory response, intestinal morphology, and ileal gene expression were assessed. ZnO improved daily gains 4 days after challenge, T3 and T4 showing intermediate values (96, 249, 170, and 157 g/d for T1, T2, T3, and T4, p = 0.035). Fecal lactobacilli were higher with T3 and T4 compared to ZnO (7.55, 6.26, 8.71, and 8.27 cfu/gFM, p = 0.0007) and T3 increased the lactobacilli/coliforms ratio (p = 0.002). T4 was associated with lower levels of Pig-MAP (p = 0.07) and increases in villus/crypt ratio (1.49, 1.90, 1.73, and 1.84, p = 0.009). Moreover, T4 was associated with an upregulation of the REG3G gene (p = 0.013, pFDR = 0.228) involved in the immune response induced by enteric pathogens. In conclusion, both plant supplements enhanced animal response in front of an ETEC F4 challenge probably based on different modes of action.

Salmonella is a common causative agent of enteric disease and is developing mechanisms of resistance to antimicrobials. Probiotics, such as bifidobacteria and lactobacilli, and prebiotic fibers are a potential alternative to counteract this pathogen as they have demonstrated effectiveness in preventing its adhesion, reducing intestinal damage, and enhancing the host immune system. Furthermore, the benefits are expected to be potentiated when these compounds are administered together. A trial was performed to evaluate the efficacy of two probiotic strains (Bifidobacterium longum subsp. infantis CECT 7210 (Laboratorios Ordesa S.L.) and Lactobacillus rhamnosus HN001, combined or not with a prebiotic containing oligofructose-enriched inulin, against Salmonella Typhimurium. Ninety-six piglets (28 days old) were distributed into 32 pens assigned to 5 treatments: one non-challenged (control diet, CTR+) and four challenged: control diet (CTR−) or supplemented with probiotics (>3 × 1010 cfu/kg each strain, PRO), prebiotic (5%, PRE), or their combination (SYN). After 1 week of adaptation, animals were orally challenged with Salmonella Typhimurium. Feed intake, weight, and clinical signs were recorded. On days 4 and 8 post-inoculation (PI), one animal per pen was euthanized, and samples from blood, digestive content, and ileal tissues were collected to determine Salmonella counts, fermentation products, ileal histomorphology, and serum TNF-α and Pig-MAP concentrations. The effect of the oral challenge was evidenced by animal performance, fecal consistency, and intestinal architecture. Regarding the experimental treatments, animals belonging to the PRO group experienced a faster clearance of the pathogen, with more pigs being negative to its excretion at the end of the study and recovering the impaired ileal villi/crypt ratio more rapidly. Animals receiving the PRE diet showed a lower intestinal colonization by Salmonella, with no countable levels (

Short and medium-chain fatty acids (SCFA and MCFA, respectively) are commonly used as feed additives in piglets to promote health and prevent post-weaning diarrhoea. Considering that the mechanism and site of action of these fatty acids can differ, a combined supplementation could result in a synergistic action. Considering this, it was aimed to assess the potential of two new in-feed additives based on butyrate or heptanoate, protected with sodium salts of MCFA from coconut distillates, against enterotoxigenic

Enterotoxigenic Escherichia coli (ETEC) K88 (F4) is a worldwide cause of severe diarrhoea in the offspring of some animal species such as weaned pigs. Preventing ETEC K88 (F4) attaching to the small intestine by natural feed ingredients interfering with its fimbria-receptor recognition could be an alternative method to reduce diarrhoea occurrence. The purpose of this study is to evaluate the ability of different common ingredients in the formulation of pig diets, and particularly different kinds of cereal brans, to attach ETEC K88 (F4) (adhesion test, AT) and to block its attaching to intestinal epithelial cells (IPEC-J2) (blocking test, BT) in vitro. For the feed ingredients, wheat, corn, oats, barley, rye, extruded full-fat soybean meal, soybean meal, as well as sweet whey powder, all demonstrated a clear affinity to attach ETEC K88 (F4) (P

Intensive farming may involve the use of diets, environments or management practices that impose physiological and psychological stressors on the animals. In particular, early weaning is nowadays a common practice to increase the productive yield of pig farms. Still, it is considered one of the most critical periods in swine production, where piglet performance can be seriously affected and where they are predisposed to the overgrowth of opportunistic pathogens. Pig producers nowadays face the challenge to overcome this situation in a context of increasing restrictions on the use of antibiotics in animal production. Great efforts are being made to find strategies to help piglets overcome the challenges of early weaning. Among them, a nutritional strategy that has received increasing attention in the last few years is the use of probiotics. It has been extensively documented that probiotics can reduce digestive disorders and improve productive parameters. Still, research in probiotics so far has also been characterized as being inconsistent and with low reproducibility from farm to farm. Scientific literature related to probiotic effects against gastrointestinal pathogens will be critically examined in this review. Moreover, the actual practical approach when using probiotics in these animals, and potential strategies to increase consistency in probiotic effects, will be discussed. Thus, considering the boost in probiotic research observed in recent years, this paper aims to provide a much-needed, in-depth review of the scientific data published to-date. Furthermore, it aims to be useful to swine nutritionists, researchers and the additive industry to critically consider their approach when developing or using probiotic strategies in weaning piglets.

Salmonella spp. is one of the worldwide leading causes of food-borne illnesses for which the inclusion of probiotics or organic acids in animal feeds can be useful control methods. Experimental models are utilized to test the efficacy of strategies against pathogens, but they exhibit limitations which may preclude finding sensible evaluation parameters. The objective of this work is to evaluate the efficacy of 2 different feed additives; a Bacillus licheniformis based probiotic and a protected sodium butyrate (SB) salt, using an experimental model of salmonellosis and, second, to explore if behavior analysis can be used as a sensible evaluation tool for additives evaluation. A total of 78 piglets weaned at 24 d, 8.3 kg BW, were used. Seventy-two were placed in 3 rooms of 8 pens (3 animals/pen) with evenly distributed treatments (n = 8): CON, control group with plain diet; PRO, plain diet with 1 kg/t of Proporc (109 cfu of B. licheniformis/kg of feed), and BUT, plain diet with 3 kg/t of Gustor BP70 (2.1 g of partially protected SB salt/kg of feed). Remaining piglets (n = 6) were separated and used as a challenge negative control. The experiment lasted 16 d. After 1 wk of adaptation, animals were challenged with 5 × 108 cfu of Salmonella Typhimurium. One pig per pen was euthanized and sampled at d 4 and 8 post-inoculation (PI). There were no significant differences among treatments for ADFI, ADG, G:F, rectal temperature, fecal consistency, pH, ammonia, short-chain fatty acids and lactic acid concentrations, cytokine TNF-α, Pig-MAP acute-phase proteins and histological parameters. However, both products were equally able to reduce colonization and shedding of Salmonella (P = 0.016 for PRO and BUT vs. CON). In addition, PRO treatment had a positive effect on behavioral displays, particularly exploring (P < 0.05 vs. CON), feeding (P < 0.05 vs. CON and BUT) and other active behaviors (P < 0.05 vs. CON and BUT) in the morning period (0830 to 1030 h). In the afternoon (1400 to 1600 h), the challenge effect was most significant. Pigs were less active after the challenge (P < 0.001), with a decrease in positive contacts (P = 0.004), exploration (P < 0.001) and feeding behaviors (P < 0.001) on d 3 PI, in comparison with before the challenge. Accordingly, many lying conducts increased at d 3 PI (P < 0.05). In conclusion, both treatments had positive effects against Salmonella, and behavior analysis appears to be a sensible tool to be considered.

The aim of the present study was to explore the evolution of piglet gut microbiota from birth to weaning. Moreover, it was hypothesized that different farm environments could condition this process. Two farms, distinct in their use of antibiotics, and 10 litters per farm were selected. A total of 100 fecal samples were obtained from the same pig of each litter on d2, d7, d14 and d21 of lactation and d14 after weaning. The DNA was extracted by using the PSP® Spin Stool DNA Kit and sequencing of the 16S rRNA gene (V3-V4 region) performed by Illumina MiSeq Platform. Bioinformatics and biostatistical analysis were performed with QIIME and the open-source software R v3.5.3. (phyloseq package). Alpha diversity was strongly affected by age (P< 0.001), with an increasing richness of species through time. Beta diversity decreased after weaning (P< 0.001), suggesting a convergent evolution among individuals. Regarding the structure of the microbiota, a clear clustering of the samples according to age was observed (P< 0.001). A progressive decrease was observed as the piglets aged for Clostridiaceae, Enterobacteriaceae, Fusobacteriaceae, Pasteurellaceae and Streptococcaceae (P< 0.001). In contrast, Lachnospiraceae (P=0.003), Lactobacillaceae (P=0.003) and Veillonellaceae (P=0.025) increased along the d7–d14 period, but decreased afterwards. Campylobacteraceae, Erysipelotrichaceae, Ruminococcaceae (P< 0.001) and Prevotellaceae (P=0.005) gradually increased with age reflecting the change from a milk-oriented microbiome towards a butyrate-producing one. Regarding the impact of the farm, differences in species richness were found and also a distinct microbial structure (ANOSIM: P=0.025) associated to changes in some particular taxonomic groups. In conclusion, during the transition from birth to weaning, the pig microbiota showed a relevant succession of microbial groups towards a more stable ecosystem better adapted to the dry feed. In this relevant early-age process differences between farms seems to have a limited impact.

29 Páginas.-- 2 Figuras.-- 3 Tablas, The present study aimed to evaluate different mates of Candida boidinii and Lactobacillus pentosus strains as starters in green table olive fermentation. Changes in fermentation characteristics as well as changes in the functional properties of the microbial exopolysaccharides (EPS) produced during the process were registered. The in vitro adhesion test demonstrated that most EPS samples could specifically attach ETEC K88. In vitro studies with porcine intestinal cells showed the improved blocking activity of the fimbria (blocking test) when the mutant strain L. pentosus 119-14MT was used alone as a starter. All EPS samples showed the ability to block receptors in the cells (exclusion test) although without differences between starter treatments. In the displacement test, EPS samples failed to remove the pathogen once attached. According to these results, L. pentosus 119-14MT, a high EPS variant, seemed to be the most effective starter improving the anti-adhesive properties of brine EPS and increasing its ability to block the ETEC K88 fimbria. These results illustrate that the anti-adhesive properties of the EPSs produced during the traditional fermentation of olives could be modulated by the use of defined starters. This opens the door to new fermentation processes aimed to produce green table olives as functional food to prevent ETEC diarrhea., The authors wish to thank Mr Chuck Simmons, a native English-speaking, retired instructor of English of the Universitat Autònoma de Barcelona, for proofreading the manuscript. This work was supported by the Spanish Government through MICIIN projects (Project AGL 2012-31924 and AGL2013-48300-R/ALI) and by scholarships from the Chinese Scholarship Council (CSC) awarded to Y. Zhu.

This study aimed to evaluate the potential of two new fat-protected butyrate or heptanoate salts to improve gut health and control post-weaning colibacillosis in weaning piglets challenged with enterotoxigenic Escherichia coli (ETEC) F4+, particularly focusing on their impact on intestinal microbiota and fermentative activity along the gastrointestinal tract (GIT). Seventy-two 21-d-old pigs were fed a plain diet (CTR) or supplemented with sodium butyrate (BUT) or sodium heptanoate (HPT), both at 0.3%. After a week of adaptation, animals were orally challenged at days 8 and 9 with 5.8 · 109 and 6.6 · 1010 cfu, respectively, and were euthanised on d 4 and d 8 post-inoculation (PI) (n = 8) to collect blood, digesta and tissue samples and characterise microbial groups, pathogen loads (qPCR), fermentation, ileal histomorphometry and immune markers. Colonic microbiota was analysed by 16S rRNA gene MiSeq sequencing. Supplementing both acid salts did not compensate clinical challenge effects nor performance impairments and neither histomorphometry nor serum biomarkers. Changes in the gastric fermentative activity were registered, BUT reducing lactic acid concentrations (day 8 PI), and with HPT fewer animals presenting detectable concentrations of propionic, butyric and valeric acids. At ileum BUT increased acetic acid concentration (day 8 PI), and both additives reduced short-chain fatty acids (SCFA) in the colon. Increases in enterobacteria and coliforms counts in ileal digesta (day 4 PI, p < 0.10) and mucosa scrapes (p < 0.05) were registered although E. coli F4 gene copies were unaffected. Regarding changes in the colonic microbiota (day 4 PI), Prevotellaceae and Prevotella were promoted with BUT supplementation whereas only minor groups were modified in HPT-treated animals. Summarising, although the pathogen loads or inflammatory mediators remained unresponsive, butyrate and heptanoate showed a significant impact on microbial fermentation along the whole GIT, being able to modify different bacterial groups at the colon. It could be hypothesised that these effects might be mediated by a carry-over effect of the changes observed in gastric fermentation, but possibly also to a better nutrient digestion in the foregut as a result of the reduced colonic SCFA concentrations.

Hypothesis: The probiotic strain Bacillus toyonensis (Toyo) may interfere with the quorum-sensing (QS) mechanisms of Escherichia coli (ETEC) K88. Three different in vitro approaches were performed. Materials and methods: Trial 1: The adhesiveness of ETEC-K88 was evaluated using porcine intestinal epithelial cells (IPEC-J2). Bacteria were grown in the ileal and the colonic sterile digesta supernatants (SN) obtained from 32 weaned piglets supplemented (TREAT) or not (CTR) with Toyo (109 CFU/g) and subsequently added to IPEC-J2. Trial 2: Using a similar model to Trial 1, E. coli was grown in media including sterile SN obtained from cultures of E. coli coincubated or not with Toyo or sterile SN from Toyo incubated or not with acyl-homoserine lactone (AHL). Trial 3: The gene expression of the fimbrial adhesin (F4) and the heat-labile enterotoxin (LT) of ETEC-K88 was also determined after different treatments. Results: The incubation of bacteria with the ileal TREAT leads to a reduction in the adhesion of E. coli (P = 0.06). As expected, ETEC-K88 grown with its own SN increased its ability to colonize the epithelium (P = 0.023), which was slightly reduced by Toyo. Gene expression results suggest that Toyo was able to reduce the expression of F4 induced by the ETEC-K88 SN (0.64-fold changes). Moreover, when ETEC-K88 is grown in the supernatant of a previous culture of Toyo with AHL or it is cocultured with Toyo in the presence of AHL, the expressions of F4 (0.61-fold changes) or LT (0.67-fold changes) were also reduced, respectively. Conclusion: These results suggest the ability of B. toyonensis to act on the QS systems of ETEC-K88 by different complex mechanisms.

Having sensitive serum biomarkers able to determine the structural changes of the small intestine suffering from bacterial digestive diseases could be a valuable tool particularly in piglets at weaning, when intestinal infections are highly prevalent. We evaluated the usefulness of three inflammatory and gut-wall-integrity biomarkers to assess the degree of intestinal histo-morphological damage in piglets. Piglets were orally challenged with Salmonella Typhimurium or enterotoxigenic Escherichia coli (ETEC) to get a variable range of response according to individual variability. Forty-eight piglets were challenged with Salmonella Typhimurium and seventy-two with enterotoxigenic Escherichia coli K88. Clinical signs and faecal score were recorded. At Days 4 and 8 post-inoculation, blood was sampled, animals euthanised and distal ileum dissected. Morphological measures were obtained from the gut tissue, and serum tumour necrosis factor-alpha (TNF-α), pig major acute-phase protein (Pig-MAP) and intestinal fatty acid-binding protein (I-FABP) were determined. Animals developed mild-to-severe diarrhoea after the challenge. When analysing the complete set of analytical results, a high correlation was found among the three serum biomarkers. The most representative morphological indicator was the villus:crypt ratio (V:C), which showed a strong negative correlation with all three biomarkers. Regression analyses between faecal score and the previous variable showed linear relations. When the range of V:C was analysed, based on the quartile distribution of each serum variable, a marked increase in their concentration was observed with greater villus damage. Summarising, the combination of I-FABP, Pig-MAP and TNF-α may be useful for determining the intestinal injury degree and barrier integrity in recently weaned pigs.

Plant ingredients contain low Ca levels and 1/3 may be bound to P phytate. Therefore, Ca has to be supplemented with animal or mineral sources, such as limestone or calcium phosphates. However, growth performance and feed intake has been reported by other authors to decrease linearly with increasing dietary Ca. To better understand the possible mechanisms underlying detrimental effect of including limestone in weanling diets, two experimental trials were performed to evaluate its possible impact on growth performance, colonic microbiota and, jejunal gene expression. In Trial 1, a total of 240 piglets (26 ± 2 d old, average body weight (BW) = 7.7 ± 1.04 kg) were distributed into 3 dietary treatments: 0% limestone, 0.8% limestone, and 1.6% limestone to provide 0.6, 0.9 and 1.2% total Ca, respectively. Feed intake and individual BW were registered during the pre-starter phase (d 0–14 post-weaning). Pigs fed diet with 1.6% of limestone (1.2% of total Ca) had lower BW, average daily gain (ADG), and gain to feed ratio (G:F) compared with pigs fed diets with 0.8 and 0% limestone (0.9 and 0.6% total Ca, respectively; P < 0.05). In Trial 2, a total of 18 pigs (28 ± 0 d old, average BW = 7.2 ± 0.24 kg) were allocated individually in cages and assigned to 2 dietary treatments: 0% limestone and 1.4% limestone to provide 0.6 and 1.2% total Ca, respectively. Piglets were fed for 14 d and then euthanized to obtain jejunum tissue for gene expression and colon digesta for 16SRNA gene microbiota analyses. Pigs fed 1.4% limestone (1.2% of total Ca) showed a higher beta-diversity and an increase on the Bacteroides genera in colon (P < 0.05). These animals also showed an up regulation of gene sets related to the cell cycle regulation, DNA and RNA transcription and inflammatory response in the jejunum. In conclusion, high levels of limestone inclusion in weaning diets, decreases growth performance without affecting feed intake in weaned pigs. Also, addition of limestone to diets for 14 d after weaning can upregulate the expression of genes related to the inflammatory response, and enlarge colonic beta-diversity with an increased Bacteroides genera.

33 Páginas.-- 6 Figutas.-- 1 Tabla.-- 3 Figuras suplementarias, This study aims to explore the biological functions of the isolated exopolysaccharides (EPSs) produced during the industrial fermentation of olives against enterotoxigenic E. coli (ETEC) K88. Exopolysaccharides were isolated from five industrial fermenters. Analysis of their monosaccharide composition by GLC revealed that the main components were glucose (27%–50%) and galactose (23%–33%) followed by rhamnose (4–23%) and arabinose (6–17%). The 1H NMR spectrum showed a very similar profile between samples, and a more in-depth analysis revealed the presence of an α-pyranose in the form of α-D-Glcp-(1→) and two different α-furanoses, with chemicals shift values, suggesting the presence of α-D-Glcf and α-D-Galf. Miniaturized in vitro tests demonstrated the ability of EPS samples to attach specifically to ETEC K88 (P < 0.05) with variable intensities. The competition test did not show the ability to block the ETEC K88 adhesion to IPEC-J2 cells; however, in the displacement test, all EPS samples were shown to effectively remove the pathogens attached to the cells (P < 0.01). These results suggest that the EPSs produced during the fermentation of table green olives could interfere with the attachment of opportunistic pathogens onto the intestinal epithelial cells. This would open the possibility of novel functional properties for this traditional Mediterranean fermented food and for the isolated EPSs as candidates for nutraceutics to be used in human and/or animal diets in the prevention and treatment of ETEC diarrhoea., The authors wish to thank Mr Chuck Simmons, a native English-speaking instructor of English at the Universitat Autonoma de Barcelona for proofreading the manuscript. This work was supported by the public research project programme of the Spanish Ministry of Economy and Competitiveness (Project AGL 2012-31924) and by scholarships from the Chinese Scholarship Council (CSC) awarded to Y. Zhu.

Probiotics have been demonstrated to be useful to enhance gut health and prevent gastrointestinal infections in humans. Additionally, some multi-strain probiotic combinations have been suggested to have greater efficacy than single strains. The objective of this study is to demonstrate the potential of a combination of the probiotic strains: Bifidobacterium longum subsp. infantis CECT 7210 (brand name B. infantis IM1®) and B. animalis subsp. lactis BPL6 to enhance gut health and to ameliorate the outcome of a Salmonella challenge using a weaning piglet model. Seventy-two 28-day-old weanling piglets, 7.7 (±0.28) kg of body-weight, were distributed in a 2 × 2 factorial design; treated or not with the probiotic combination and challenged or not with the pathogen. Animals were orally challenged after an adaptation period (Day 8) with a single dose (5 × 108 cfu) of Salmonella Typhimurium. One animal per pen was euthanized on Day 12 (Day 4 post-inoculation [PI]) and Day 16 (Day 8 PI). All parameters responded to the challenge and 4 deaths were registered, indicating a severe but self-limiting challenge. Improvements registered in the challenged animals due to the probiotic were: increased voluntary feed-intake (P probiotic × challenge = 0.078), reduced fecal excretion of Salmonella (P = 0.028 at Day 1 PI and P < 0.10 at Days 3 and 5 PI), decreased rectal temperature (P probiotic × day = 0.048) and improvements in the villous:crypt ratio (P probiotic × challenge < 0.001). Moreover, general probiotic benefits were observed in both challenged and non-challenged groups: decreased diarrhea scores of the PI period (P = 0.014), improved fermentation profiles on Day 8 PI (increased ileal acetic acid [P = 0.008] and a tendency to lower colonic ammonia concentrations [P = 0.078]), stimulation of intestinal immune response by increasing villous intraepithelial lymphocytes (P = 0.015 on Day 8 PI) and an improved villous:crypt ratio (P = 0.011). In conclusion, the multi-strain probiotic had a positive effect on reducing pathogen loads and alleviating animals in a Salmonella challenge. In addition, enhanced gut health and immunity was recorded in all animals receiving the probiotic, indicating an improvement in the post-weaning outcome.

Probiotics have been demonstrated to be useful to enhance gut health and prevent gastrointestinal infections. The objective of this study is to demonstrate the potential of the probiotic strain Bifidobacterium longum subsp. infantis CECT 7210 (B. infantis IM1) to prevent and fight intestinal disease by using a Salmonella Typhimurium (Trial 1) or an enterotoxigenic Escherichia coli K88 (Trial 2) oral challenge in a weaning piglet model. Seventy-two piglets were used in each trial. After an adaptation period, animals were orally challenged. One animal per pen was euthanized at Days 4 and 8/9 (Trial 1/Trial 2) post-inoculation (PI). Animal performance, clinical signs, pathogen excretion, fermentation, immune response, and intestinal morphology were evaluated. In Trial 1, most parameters responded to the challenge, whereas, in Trial 2, effects were much milder. Consistent effects of the probiotic were detected in both experiments: Reduction of pathogen excretion (P = 0.043 on Day 3 PI, Trial 1) or ileal colonization (33% reduction of animals with countable coliforms; P = 0.077, Trial 2); increases in intraepithelial lymphocytes (P = 0.002 on Day 8 PI in Trial 1, P = 0.091 on Day 4 PI in Trial 2), and improvement of the fermentation profile by increasing butyric acid in non-challenged animals [P challenge × probiotic (interaction) = 0.092 in Trial 1 and P = 0.056 in Trial 2] concomitant with an enhancement of the villus:crypt ratio on Day 8/9 PI (P interaction = 0.091 for Trial 1 and P = 0.006 for Trial 2). Challenged animals treated with the probiotic showed reduced feed intakes (P interaction = 0.019 in Trial 1 and P = 0.020 in Trial 2) and had lower short-chain fatty acid concentrations in the colon (P interaction = 0.008 in Trial 1 and P = 0.082 in Trial 2). In conclusion, this probiotic demonstrated potential to reduce the intestinal colonization by pathogens and to stimulate local immune response. However, effects on feed intake, microbial fermentation, and intestinal architecture showed a differential pattern between challenged and non-challenged animals. Effects of the probiotic intervention were dependent on the structure of the ecosystem in which it was applied.

Some plant extracts, have been demonstrated to interfere with the microbial metabolism of several pathogenic bacteria. Within this antimicrobial properties it has been described the potential to inhibit or destroy biofilms or to interfere in quorum-sensing (QS) systems. However, to our knowledge, no study exploring this potential of wheat-bran (WB) has been published. The purpose of the present study is to evaluate the anti-biofilm activity of WB against a cow mastitis strain of Staphylococcus aureus and also its possible interference with bacterial QS systems. The potential of inhibition and destruction of the biofilm was studied by different in vitro assays. Also, we tested the ability of WB to interfere in bacterial QS by degrading acyl-homoserine lactones (AHL) as one of the most studied QS signal molecules for Gram-negative bacteria. The soluble extract of WB at 0.5% showed anti-biofilm activity, inhibiting biofilm formation and also destroying it. Similarly, the > 300 kDa fraction from WB had significant anti-biofilm activity in both in vitro assays. The WB also showed a potential to interfere with bacterial QS systems, as it was demonstrated to contain certain lactonase activity able to reduce AHL concentration in the medium. The present study reveals two additional beneficial properties of WB extract never explored before, which may be related to the presence of defence compounds in the plant extract able to interfere with microbial biofilms and also QS systems.

The prebiotic lactulose, a probiotic strain of Lactobacillus plantarum (L. plantarum) and a synbiotic combination of these two agents were evaluated as growth promoters in 25–39-day old commercial weaning pigs. Ninety-six weaning pigs were allocated into 32 pens, taking initial weight into account, and distributed into four groups as follows: a control diet (CTR), the same diet supplemented daily with L. plantarum (109 CFU/mL sprayed on top; 20 mL/pig) (LPN); 10 g/kg lactulose (LAC) or a combination of both treatments (SYN). At day 14, eight piglets from each group were euthanized and proximal colon digesta was sampled for luminal pH, short-chain fatty acids (SCFA) and lactic acid concentrations. Deoxyribonucleic acid was extracted from colonic digesta and the microbial community was profiled by terminal restriction fragment length polymorphism analysis (T-RFLP) and qPCR. Blood urea nitrogen (BUN) and acute-phase proteins (Pig-MAP) were measured. Lactulose treatment (LAC) improved feed intake (P

Casein glycomacropeptide (CGMP), a glycoprotein originating during cheese manufacture, has shown promising effects by promoting the growth of some beneficial bacteriain vitro, although its activity has not been well explored. The present study was designed to evaluate the effects of CGMP against enterotoxigenicEscherichia coli(ETEC) K88in vitro(Trial 1) andin vivo(Trial 2). In Trial 1, increasing concentrations of CGMP (0, 0·5, 1·5 or 2·5 mg/ml) were tested regarding its ability to block the attachment of ETEC K88 to ileal mucosa tissues obtained from piglets. Increasing the concentration of CGMP resulted in a gradual decrease in ETEC K88 attachment to the epithelial surface. In Trial 2, seventy-two piglets were distributed in a 2 × 2 factorial combination including or omitting CGMP in the diet (control dietv.CGMP) and challenged or not with ETEC K88 (yesv.no). Inclusion of CGMP increased crude protein, ammonia and isoacid concentrations in colon digesta. CGMP also increased lactobacilli numbers in ileum and colon digesta, and reduced enterobacteria counts in mucosa scrapings and the percentage of villi withE. coliadherence measured by fluorescencein situhybridisation. The inclusion of CGMP in the diets of challenged animals also prevented the increase of enterobacteria in ileal digesta. We can conclude that CGMP may improve gut health by diminishing the adhesion of ETEC K88 to the intestinal mucosa, by increasing the lactobacilli population in the intestine and by reducing the overgrowth of enterobacteria in the digestive tract of piglets after an ETEC K88 challenge.

The main objective of this work was to investigate the potential of a microencapsulated mixture of organic acids (formic and lactic acid) added to the feed of fattening pigs to reduce the prevalence and shedding of Salmonella when arriving to the abattoir. Two trials were performed. In Trial 1, 24 fattening pigs received one of three diets: a control diet, the same diet including lactic and formic acid (0.4% each) (non-protected blend; NPB), or a lipid microencapsulated blend (0.14% each acid) (protected blend; PB). After 10 days, digesta samples from various parts of gastrointestinal tract were taken. No changes were detected either in pH, total short chain fatty acids (SCFA), lactic/formic acid concentrations in any of the sections studied or in caecal lactic acid bacteria or enterobacteria. In Trial 2, 261 pigs from a commercial farm were distributed between the three previous diets in the five weeks before slaughtering. At the abattoir, no change was detected in caecal pH, but increased concentrations (p < 0.05) of formic and lactic acid were found with the NPB and PB treatments. The total SCFA concentration was higher with the PB compared to the control diet (p = 0.002) with a lower percentage of branched chain fatty acids. Both acidified diets decreased enterobacteria in the caecum but did not modify lactobacilli. NPB treatment decreased Salmonella seroprevalence (p < 0.001). A significant (p < 0.05) increase in Salmonella faecal shedding was found related to the stress previous to slaughter with the control group and PB, but not in the NPB group. More studies will be needed to confirm the usefulness of protected acids to prevent Salmonella prevalence and shedding at the abattoir.

The fermentation kinetics of both fibre choice and maize particle size were studied in vitro from the ileal contents of cannulated pigs given five different experimental diets. Additionally, in vitro batch systems were used to study the quantitative effect of fibre choice and maize particle size on Salmonella Typhimurium growth. Freeze-dried ileal effluents obtained from five cannulated pigs given the five experimental diets in a latin square were used as incubation substrates. The experimental diets consisted of: (1) a standard maize-based diet (ST) where all the ingredients were milled to pass through a 2.5-ml screen, (2) the same diet with the maize ground using a 4.0-ml screen (CG), (3 and 4) two diets in which the maize was partially replaced by (3) sugar beet pulp (80 g/kg; BP) or (4) wheat bran (100 g/kg; WB), or (5) a combination of 80 g sugar beet pulp and 100 g wheat bran/kg (diet HF). Results showed that substrate from BP and HF generated more gas than substrate from ST (P vs. 125.3 mg/l, P=0.014) and the highest purine bases concentration (0.28 μmol/ml vs. 0.22 μmol/ml, P=0.009). None of the substrates showed any inhibitory effect on Salmonella growth when the batch systems were inoculated with S. Typhimurium. In summary, under the in vitro conditions used, the changes in hindgut fermentation promoted by the inclusion of different fibrous ingredients or by different maize particle size did not produce an inhibitory effect on the growth of S. Typhimurium.

The objective of this study was to evaluate the influence of different feed withdrawal and lairage times prior to slaughter on the gastrointestinal tract (GIT) weight and on the fermentation pattern and numbers of Enterobacteriaceae in the cecum of pigs. A total of 72 finishing pigs (6 pens, 12 animals each) were included in the study and were sent to the slaughterhouse on three consecutive days. On each day pigs from two pens were deprived of feed for either 2 or 12 h before leaving the farm. Pigs from each of the two pens were divided in sub-groups (4 pigs each) on the moment of loading onto the truck and followed the same distribution at the slaughterhouse. Once there, each sub-group corresponding to the same feed withdrawal time was held in different holding pens for 0, 5 or 10 h before slaughter. The weight of the gastro-intestinal tract (GIT) was determined and cecal content was collected to evaluate pH, short chain fatty acids (SCFA) and ammonia (NH 3 ) concentrations as well as lactobacilli and Enterobacteriaceae numbers. Total GIT weight decreased as feed withdrawal ( P P P P = 0.0001) (ranging from 5.79 to 6.79) in parallel to a decrease in the total SCFA concentration ( P P P 3 concentration showed an increase with withdrawal ( P P P P P P P Enterobacteriaceae numbers increased with both, feed withdrawal ( P P The previous to slaughter increase in feed withdrawal and lairage times decreased the GIT weight but involved changes in the gut microbial ecosystem caused through changes in the fermentation pattern that lead to the increase of Enterobacteriaceae numbers. This increase could represent a higher risk of carcass contamination by the enteropathogens of this group such as Salmonella . However this potential risk should be further investigated.

The aim of this study was to evaluate in vitro the influence of fermentable carbohydrates on the activity of porcine microbiota and survival of Salmonella Typhimurium in a batch culture system simulating the porcine hindgut. The carbohydrates tested were xylooligosaccharides, a mixture of fructooligosaccharides/inulin (FIN), fructooligosaccharides (FOS), gentiooligosaccharides (GEO) and lactulose (LAC). These ingredients stimulated the growth of selected Bifidobacterium and Lactobacillus species in pure cultures. In batch cultures, the carbohydrates influenced some fermentation parameters. For example, GEO and FIN significantly increased lactic acids compared with the control (no added carbohydrate). With the exception of LAC, the test carbohydrates increased the production of short-chain fatty acid (SCFA) and modified SCFA profiles. Quantitative analysis of bacterial populations by FISH revealed increased counts of the Bifidobacterium group compared with control and, with exception of FOS, increased Lactobacillus, Leuconostoc and Weissella spp. counts. Salmonella numbers were the lowest during the fermentation of LAC. This work has looked at carbohydrate metabolism by porcine microbiota in a pH-controlled batch fermentation system. It provides an initial model to analyse interactions with pathogens.

The efficacy of a commercial source of mannanoligosacharides (BM), organic zinc (BP), or their combination to enhance performance, gastrointestinal health, and immune response in weaned pigs was evaluated. A total of 128 piglets, weaned at 20 +/- 2 d, were housed in 32 pens. Animals received 1 of 4 dietary treatments: a control diet (CT) to which 0.2% of BM, 80 mg/kg of Zn as BP, or both additives (BMP) were added. The experiment lasted for 5 wk including a prestarter period of 2 wk and a starter period of 3 wk. Body weight was recorded and daily feed intake was calculated. Fecal consistency was monitored for the first 21 d. After 2 wk, 32 animals were killed, digesta samples from the stomach, ileum, and cecum were collected, and pH and the short-chain fatty acid profile were determined. Microbiological counts for enterobacteria and lactobacilli were evaluated using quantitative PCR. Histological parameters in the jejunum and immunoglobulin concentrations in serum and ileal digesta were also measured. Both additives improved G:F during the starter period (0.63, 0.69, 0.67, and 0.68 for CT, BM, BP, and BMP, respectively; P < 0.04). Mean fecal score values for the first 21 d were improved by BM and BP, showing decreased values compared with the CT diet (1.22, 0.89, 0.87, and 1.06 for CT, BM, BP, and BMP, respectively; P = 0.002). The addition of BM decreased enterobacteria counts in the jejunum (9.13, 8.05, 8.87, and 7.89 log 16S rRNA gene copies/g of matter for CT, BM, BP, and BMP, respectively; P = 0.05). Empty ileal weight, defined as the segment including the continuous Peyer's patch, tended (P = 0.08) to increase with BP treatment (8.9, 9.6, 11.9, and 10.3 g/kg of BW for CT, BM, BP, and BMP, respectively). Crypt depths in the jejunum were lower in animals fed the combination of the additives (BPM) compared with those fed the control diet (281 vs. 235; P < 0.03). No significant differences were registered in pH, short-chain fatty acids, or serum and ileal immunoglobulin concentrations. The results suggest that the use of BM or BP can improve the efficiency of gain during the starter period.

An experiment was designed to monitor the changes in caecal microbiota associated with early weaning. Twelve piglets (20+/-2 days) from six different litters were selected from a commercial source. For the two experimental groups, one animal from each litter was weaned onto a post-weaning diet (W) and the other remained with the sow (S). After 1 week, animals were sacrificed and caecal samples taken. Microbial counts for total bacteria, enterobacteria and lactobacilli populations were determined by quantitative PCR using SYBR Green dye. Microbial profiles were assessed by terminal restriction fragment length polymorphism (t-RFLP). Weaning promoted an increase in the enterobacteria:lactobacilli ratio (0.27 versus 1.67 log/log 16S rRNA gene copy number, P=0.05). Total bacteria and richness of the caecal microbial ecosystem (number of peaks) were similar in both experimental groups (49.3 for S and 53.4 for W, respectively, P=0.22), although the band patterns were clearly grouped in two different clusters by dendogram analysis. Weaning was also associated with a decrease in crypt density, an increase in mytotic index and a decrease in the number of goblet cells. A reduced immunological response was also observed and was manifested by an increase in intraepithelial lymphocytes and lymphocyte density in the lamina propria. Weaning appears to be critical in the establishment of the caecal microbiota in pigs with important changes, particularly in microbial groups and in caecal mucosal architecture.

An experiment was designed to study the effect of corn physical structure and different types of dietary fibre on pig gut microbiota during growing. Ninety-six animals (15 ± 2.2 kg) were assigned to four different diets: a control diet (CT), a diet with coarse ground corn (CC), and other two with 8% of sugar beet pulp (BP) or 10% of wheat bran (WB). Thirty-two animals (8 per diet) were sacrificed on days 7, 21 and 42 to measure microbial activity in digesta. Other 8 pigs were sacrificed on day 0 before receiving the diets. Enterobacteria and lactobacilli population in colon were quantified by qPCR. With all diets enterobacteria diminished with time and were lower than lactobacilli. Expressed as lactobacilli:enterobacteria ratio we found differences between diets at day 7 and 21 that had disappeared at day 42. Purine bases (PB) concentration indicated an adaptation of the microbiota with time which was reflected in an increasing content in the caecum and a decreasing content in the colon. Time of adaptation of microbiota was also shown by xylanase and cellulase activities that were not detected until day 7 and 42, respectively. Results obtained shows that pig gut microbiota need a relative long time to adapt after a change of diet that could last up to 6 weeks. We were not able to detect a clear effect of dietary fibre content on this process.

The effect of different types of fibre on the intestinal digesta and microbial diversity was evaluated in growing pigs. The animals were fed during forty-two days with one of four experimental diets including a control group, a diet enriched with resistant starch type I, by coarse ground corn, and other two diets containing sugar beet pulp (8%) and wheat bran (10%) (as sources of soluble and insoluble non-starch polysaccharides respectively). Body weight (BW) and average daily feed intake (ADFI) were assessed at day 7, 21 and 42. Luminal samples of digesta from proximal colon were collected to analyse water retention capacity (WRC), concentration of short-chain fatty acids (SCFA) and the microbial diversity. Overall, animals fed with higher amounts of non-digestible carbohydrates had lower ADFI and BW. Sugar beet pulp provoked an increase of WRC, a higher concentration of SCFA, and a more stable microbial diversity throughout the experimental period. On the other hand, animals fed with wheat bran presented a lower bacterial diversity.

The aim of this work was to evaluate the influence of a coarse ground cereal and two fibrous ingredients incorporated in the diet on the physicochemical properties of digesta and productive parameters of pigs during the first stage of the growing period. A total of 96 pigs (initial body weight, BW, 15 kg) were distributed into four experimental treatments: the control diet (CT) consisted of corn, barley and soybean meal milled to pass through a 2.5 mm screen; the coarse corn diet (CC) was prepared by milling the corn to a coarser particle size (4.0 mm screen); the sugar beet pulp diet (SBP) and the wheat bran diet (WB) were prepared by replacing some of the corn for sugar beet pulp (80 g/kg) or wheat bran (100 g/kg) respectively, in order to contain a higher amount of non-starch polysaccharides (NSP). Three experimental periods were considered (7, 21 and 42 days) during which body weight (BW) and voluntary feed intake were assessed. At the end of each period eight animals per diet were slaughtered. Weight of the gastrointestinal tract and its compartments (full and empty) was recorded and the contents were sampled. Digesta samples were analysed for water concentration, water retention capacity (WRC), ammonia and short-chain fatty acids concentration (SCFA). Histological study of the proximal colon tissues was also performed. In general, the different parameters evaluated showed differences between the experimental periods, but few interactions were recorded. Animals fed CC, SBP and WB diets presented a lower feed intake (P ≤ 0.009) compared to CT fed animals. Compared to the control diet, coarse grinding of corn provoked an increase in the colonic digesta content (P = 0.032). Similarly, animals fed the SBP diet compared to CT animals, showed an increase in the contents (P = 0.009) of hindgut, and in the concentration of water in the digesta (P ≤ 0.011). Compared to CT diet the SBP diet lowered the ammonia concentration in the hindgut contents (P ≤ 0.045) and increased the concentration of SCFA in the distal colon (P ≤ 0.025). Animals fed the SBP diet also showed a lower number of lymphoid nodes in the colonic mucosa compared to the other diets (P ≤ 0.043). Minor modifications were observed associated with the incorporation of wheat bran in the diet, but colonic water from WB fed animals showed a tendency to increase cytolytic capacity. The results confirm major changes in the voluntary intake and physicochemical properties of digesta as affected by the incorporation in the diet of a fibrous ingredient or coarse grinding of cereals.

A total of 32 pigs of 15+/-0.38 kg body weight were fed for 6 weeks one of four diets differing in their source of dietary fibre. FISH was used to quantify the main bacterial groups in the pig gut using the following probes: Eub338, Bac303, Rfla729, Rbro730, Erec482, Fprau645, Prop853, Str493 and Lab158. FISH counts revealed important differences at four sites along the pig gastrointestinal tract, but we were unable to show differences related to diets. Stomach and jejunal samples gave total bacterial counts of 0.1-5.3 x 10(8) g(-1) of contents. In the stomach, streptococci and lactobacilli were predominant, and the clostridial cluster IX group was abundant (14-41% of total bacterial count). Clostridial cluster IX bacteria were present elsewhere in the gastrointestinal tract at 1-8%. The other groups were generally more abundant in the proximal colon and rectum: Bacteroides/Prevotella (5-10%), clostridial cluster XIVa (10-19%), and cluster IV relatives of Faecalibacterium prausnitzii (1-4%) and ruminococcus (4-10%). Restriction fragment length polymorphism profiles showed changes related to diet, with pigs fed wheat bran having the lowest richness of all diets (P=0.008).

We evaluated the effects of 3 additives, sodium butyrate (AC), avilamycin (AB), and a combination of plant extracts (XT), on the productive performance and the intestinal environment of the early-weaned pig. The XT was a standardized mixture with 5% (wt/wt) carvacrol (from Origanum spp.), 3% cinnamaldehyde (from Cinnamonum spp.), and 2% capsicum oleoresin (from Capsicum annum). Pigs (n = 32) weaned at 18 to 22 d of age with an initial BW of 6.0 +/- 0.10 kg were allocated to 8 pens that, in turn, were allocated to 4 treatments. The treatments included a basal diet (CT) or the basal diet supplemented with 0.3% of AC, 0.04% of AB, or 0.03% of XT. Productive performance was determined during the initial 14 d postweaning. On d 19 and 21 of the experiment, the pigs were killed to allow collection of digesta and intestinal tissue to evaluate variables indicative of aspects of the gastrointestinal environment. Treatments AB and AC improved G:F (P = 0.012 and 0.003, respectively) compared with the CT. Butyrate included in the diet was only detected in the stomach but not in cranial jejunum. When compared with CT, AC produced a lower ileal starch digestibility (P = 0.002) and a lower whole-tract OM and starch digestibility (P = 0.001 and 0.003, respectively), related to a lower VFA concentration in the cranial colon (P = 0.082) and a numerically reduced branched VFA percentage in the rectum. The AB treatment diminished propionate production in caudal colon (P = 0.002) and rectum (P = 0.012) compared with CT. The AC group exhibited deeper crypt depth in the jejunum without variations in villus height compared with CT (P = 0.042). The AC and AB groups also increased goblet cell presence in the colon (P = 0.001 and 0.032, respectively). On the other hand, AB and XT diminished intraepithelial lymphocytes in the jejunum (P = 0.003 and 0.034, respectively). The XT increased lymphocyte presence in the colon (P = 0.003). These results show the important influence of AB and AC on productive performance and on pig gut dynamics. The intestinal modifications observed for AB and AC compared with CT suggest distinct modes of action for each additive.

An experiment was designed to evaluate the effects of 3 different additives on the gastrointestinal microbiota of early-weaned pigs. Early-weaned (18 to 22 d; n = 32) pigs (6.0 +/- 0.10 kg of BW) from 8 litters were randomly distributed into 8 pens. Each pen was assigned 1 of 4 dietary treatments: a prestarter or control diet, the control diet with 0.04% avilamycin (AB), with 0.3% sodium butyrate, or with 0.03% plant extract mixture (XT; standardized mixture with 5% (wt/wt) carvacrol extracted from Origanum spp., 3% cinnamaldehyde extracted from Cinnamonum spp., and 2% capsicum oleoresin from Capsicum annum). At the end of the experimental period, 8 pigs per treatment were killed, and samples of their intestinal content were taken. The total bacterial load along the gastrointestinal tract (GIT; stomach, jejunum, cecum, and distal colon) and the lactobacilli and enterobacteria in the jejunum and cecum were measured by quantitative PCR. The total microbial counts along the GIT did not differ among the diets, but there was an increase in the lactobacilli:enterobacteria ratio in the cecum of the piglets on the XT diet (P = 0.003). Restriction fragment length polymorphism of the PCR-amplified V3, V4, and V5 regions of the 16S rDNA gene showed changes in the structure of the microbial community in the jejunum. Dendrograms grouped animals by diets; control with 0.3% sodium butyrate was the treatment that promoted the biggest changes in the microbial ecosystem, followed by AB and then XT. Biodiversity increased when using additives compared with the control diet (P = 0.002). Microbial metabolic activity along the hindgut was studied using the concentration of purine bases and carbohydrase activities. Different patterns for purine bases were observed between diets (diet x intestinal section, P = 0.01). The control diet reached a maximum purine base concentration at the end of the colon, whereas that of the AB diet was reached at the cecum. We could not detect any cellulase or xylanase activities in animals of this age. Appreciable amylase and amylopectinase activities were found, but they did not differ between diets. The results suggest that the effects of avilamycin, butyrate, or the plant extract would not be related to a reduction in the number of total bacteria inhabiting different sections of the GIT but rather to changes in the ecological structure and metabolic activity of the microbial community.

The present study evaluated in vitro the effect of technological processes on four cereal grains: maize, wheat, barley and oats and one legume grain: green seeded pea. Processing included extrusion, cooking and grinding (0.8 and 3.0 mm opening), and the parameters evaluated were starch hydrolysis, content of soluble non-starch polysaccharides (NSP) and physicochemical properties: solubility of dry matter (DM), viscosity of the supernatant and water retention capacity (WRC). All samples were submitted to an in vitro digestion that consisted of two steps, gastric (G; pepsin; pH 2; 4 h) and small intestine (SI; pancreatin; pH 7; 4 h) simulation. Prior to incubation, particle size was evaluated in all samples by hand sieving and calculated on a weight basis. Total starch and total NSP analyses were performed on raw samples (0.8 mm). Hydrolysis of starch was evaluated by measuring glucose in the supernatant during the course of SI incubation. Soluble NSP were evaluated in the supernatant at the end of SI incubation. Physicochemical properties were evaluated after G and SI incubation. In raw ingredients (0.8 mm) the hydrolysis of starch at the end of SI incubation (ratio between glucose released in the sample and glucose released in the sample gelatinized) was lower in peas (0.45) than in cereal grains (0.69 in maize; 0.95 in wheat; 0.82 in barley and 0.92 in oats). The highest amounts of soluble NSP were found in barley (19.8 g/kg DM) and oats (13.8 g/kg DM), and were associated with higher viscosities. The WRC in raw samples was positively related to the amount of total NSP after both G (r2 = 0.847, P Technological processing increased starch hydrolysis and solubilization of NSP in all ingredients. The extent of the effect was dependent on the ingredient as well as on the severity of the technological process. Component monosaccharides of soluble NSP were differently affected, while glucose was increased by all the processes, arabinose and xylose required the more extreme conditions of extrusion. Contrary to raw samples, WRC of heat treated ingredients showed a negative relation with total NSP content of the sample after G incubation, which turned out to be a positive relation after SI incubation. It is concluded that technological processing of ingredients promotes higher starch hydrolysis, increases in the amount of soluble NSP and modifications in the physicochemical properties depending on the nature of the feed ingredients.

Jejunum digesta samples were taken from weaning pigs in order to evaluate real-time PCR (qPCR) as a method for quantifying pig gut bacteria. Total bacteria, lactobacilli and enterobacteria were quantified by qPCR and the results were compared with those obtained with traditional methods: 4',6-diamidino-2-phenylindole (DAPI staining) for total bacteria, selective culture for lactobacilli and enterobacteria. Real-time PCR showed higher values in terms of 16S rRNA gene copies than DAPI counts or CFU. Despite the differences, the lactobacilli:enterobacteria ratio was similar between methods (2.5 +/- 0.58 for qPCR and 3.1 +/- 0.71 for selective culture, P = 0.39). Possible reasons for the higher PCR counts are discussed considering both an overestimation with PCR by quantification of dead bacteria or free DNA and also an underestimation with conventional methods. Inherent differences in the pre-treatment of the samples could partially explain the discrepancies observed. Regardless of the numerical differences between methods, values obtained by qPCR and traditional methods showed a significant correlation for lactobacilli and total bacteria. In the light of these results, real-time PCR seems a valid method to quantify microbial shifts in the gastrointestinal tract.