Efficacy of medium-chain fatty acid salts distilled from coconut oil against two enteric pathogen challenges in weanling piglets

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 × 10 CFU) or enterotoxigenic Escherichia coli (ETEC) F4 (1.5 × 10 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

Management and Feeding Strategies in Early Life to Increase Piglet Performance and Welfare around Weaning : a review

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)

Terminal restriction fragment length polimorphism (T-RFLP) : una herramienta útil para valorar la dinámica y la diversidad de la población microbiana en aves y cerdos

El ecosistema intestinal es un medio complejo y dinámico que puede verse afectado puntualmente por numerosos factores propios del individuo pero también por numerosos factores ambientales o externos (como puede ser la dieta). La necesidad de conocer de una forma rápida y reproducible cómo es la dinámica de la población bacteriana, así como su estructura está siendo posible gracias al desarrollo de técnicas moleculares aplicadas como DGGE (Denaturing Gradient Gel Electrophoresis)1, TGGE (Temperatura Gradient Gel Electrophoresis)2, SSCP (Single Strand Conformation Polimorphism)3 y T-RFLP (Terminal Restriction Fragment Length Polimorphism)4. Estas técnicas moleculares aprovechan la característica de universalidad del gen que codifica para el 16S ARNr, presente en prácticamente la totalidad de las especies bacterianas. La presencia de secuencias altamente conservadas hace posible el diseño de primers o cebadores que permitan la amplificación mediante PCR del total de la población bacteriana de la muestra. Son las regiones variables (polimórficas) del gen que codifica el 16S ARNr las que se aprovechan para la diferenciación. En el T-RFLP, estas diferencias entre especies o géneros bacterianos se ponen de manifiesto tras la digestión con endonucleasas de restricción del producto de PCR marcado. Únicamente el fragmento del extremo terminal será visible por electroforesis capilar gracias al mareaje fluorescente de uno de los cebadores. El perfil que se genera (electroferograma) nos permite obtener información del número de fragmentos (riqueza), de su tamaño en pares de bases (bp) (para una posible inferencia de la especie bacteriana presente) y de su altura (lo que puede orientar sobre la importancia de un determinado grupo bacteriano dentro de una muestra), siempre sin olvidar las desviaciones inherentes propias de la PCR. Es posible también calcular la frecuencia de detección de un determinado pico (especie) respecto al total de muestras así como construir con ayuda de programas informáticos matrices de similitud o dendogramas. Empleando herramientas disponibles en la red como: TAP-tRFLP del software Ribosomal Database Project II o ISPaR del software MiCA (Microbial Community Analysis) es posible hacer una asignación teórica de la especie o género bacteriano a cada uno de los picos en base a las secuencias depositadas por otros autores. Sin embargo un mismo pico puede tener su origen en distintas especies. Disponer alternativamente de secuencias propias obtenidas tras la clonación del 16S ARN de muestras de nuestros animales nos permite hacer una asignación teórica más firme, considerando únicamente aquellas especies mayoritarias encontradas

Behavioural changes in weaned piglets orally challenged with Escherichia coli F4 and supplemented with in-feed protected acid salts

Altres ajuts: acords transformatius de la UABImprovement of the intestinal health of piglets at weaning is a principal objective in pig farming in terms of performance and welfare benefits. Early indicators of disease are indispensable for evaluating animal health and the efficacy of interventions such as feed additive supplementation. This study evaluates behavioural changes in weaned piglets that are orally challenged with enterotoxigenic Escherichia coli F4, and which receive or not two different in-feed additives. Two independent trials were performed with early-weaned piglets in pens of three animals, which were fed a plain diet (N = 32) or one supplemented with sodium butyrate (N = 16; Trial 1) or sodium heptanoate (N = 16; Trial 2) and protected with coconut distillates. After one week of adaptation, piglets were challenged with a single oral dose of E. coli F4 (minimum 1.4 x109 cfu). Scan-sampling was used to evaluate individual behaviour (location in the pen, postures, contact with pen mates, and activities) on the day before (d-1) and two days post-inoculation (d+2 and d+3) at 2-min intervals. Behaviours were recorded in mornings (8 am to 10 am) and afternoons (4 pm to 6 pm). Faecal consistency was also recorded for each animal. Diarrhoea peaked at d+ 2. Regarding behaviour, on d+ 2 there was greater frequency of the animals lying inactively under the heat lamp, in contrast to d-1 when they were more frequently present in the feeder, standing and active (P < 0.05). Around the feeder, standing and active behaviour increased at d+ 3, especially in the afternoon (PDayxTime of day < 0.05). Piglets fed sodium heptanoate spent less time around the feeder (P < 0.05). The weight of the animal at weaning was also observed to have an impact on the effect of time or diet on behaviour. Medium size piglets spent more time lying with pen mates in the afternoon (PSizexTime of day < 0.01) and the smallest piglets increased their feeding behaviour when receiving the supplemented diets (PSizexDiet < 0.05). In conclusion, a lethargic response among piglets after the E. coli F4 challenge was evidenced, this response being slightly modified by the supplementation of in-feed additives and the size of the animals. These results are evidence of the potential of behavioural indicators as a useful tool to assess the health status of piglets at weaning and their responses to in-feed supplementation, but they should be regarded with caution before any transfer to farm conditions due to the limitations of experimental models

Screening the ability of natural feed ingredients to interfere with the adherence of enterotoxigenic Escherichia coli (ETEC) K88 to the porcine intestinal mucus

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,0·001). In the BT, WB, CGMP and locust bean (LB) reduced the number of ETEC K88 attached to the intestinal mucus (P,0·001). For WB and LB, fractionation based on their carbohydrate components was subsequently carried out, and each fraction was evaluated individually. None of the WB fractions reduced the adhesion of ETEC K88 to the mucus as did the original extract, suggesting that a protein or glycoprotein could be involved in the recognition process. With regard to the LB fractions, the water-extractable material reduced the adhesion of ETEC K88 (P,0·001) to the mucus similar to the original extract (P,0·001), indicating, in this case, that galactomannans or phenolic compounds could be responsible for the recognition process. In conclusion, among the FI screened, the soluble extracts obtained from WB, LB and CGMP exhibited the highest anti-adhesive properties against ETEC K88 in the BT. These results suggest that they may be good candidates to be included in diets of weaned piglets for the prevention of ETEC K88-induced diarrhoea

Evaluation of dietary supplementation of a novel microbial muramidase on gastrointestinal functionality and growth performance in broiler chickens

This study was conducted to assess the effect of dietary supplementation of Muramidase 007 to broiler chickens on gastrointestinal functionality, evaluating growth performance, apparent ileal digestibility, intestinal histomorphology, vitamin A in plasma and cecal microbiota. A total of 480 one-day male chicks (Ross 308) were distributed in 16 pens allocated in 2 experimental diets: the control diet (CTR) without feed enzymes, coccidiostat or growth promoters, and the experimental diet (MUR): CTR supplemented with 35,000 units (LSU(F))/kg of the Muramidase 007. Digesta and tissue samples were obtained on days 9 and 36 of the study. A lower feed conversion ratio was observed in the MUR treatment. Apparent ileal digestibility of DM, organic matter and energy were improved by Muramidase 007. It was also observed that MUR improved digestibility of total fatty acids, mono-unsaturated fatty acids and poly-unsaturated fatty acids, and content of vitamin A in plasma at day 9 (P < 0.05). Histomorphological analysis of jejunum samples revealed no differences in the villus height or crypt depth; but a higher number of goblet cells and intraepithelial lymphocytes at day 36 with MUR. No differences were observed in plate counts of enterobacteria or Lactobacillus along the gastrointestinal tract, neither on the cecal short-chain fatty acids. An statistical trend was observed for reduction of cecal clostridia at day 9 for MUR. Analysis of cecal microbiota structure by 16S rRNA gene sequencing revealed relevant changes correlated to age. At day 9, broilers receiving MUR showed decreased alpha diversity compared to CTR that was not detected at day 36. Changes in specific taxonomic groups with an increase in Lactobacillus genus were identified. In conclusion, evaluation of the variables in this study indicates that dietary Muramidase 007 contributes to improve feed conversation ratio and gastrointestinal function in broiler chickens. Effects could have been mediated by slight shifts observed in the intestinal microbiota. More studies are guaranteed to fully understand the mechanisms involved.info:eu-repo/semantics/publishedVersio

Effects of Bifidobacterium longum Subsp. infantis CECT 7210 and Lactobacillus rhamnosus HN001, Combined or Not With Oligofructose-Enriched Inulin, on Weaned Pigs Orally Challenged With Salmonella Typhimurium

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 × 10 10 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 (<3 cfu/g) in any of the analyzed samples, and an augmented immune response suggested by serum Pig-MAP concentrations. Treatments including the prebiotic (PRE and SYN) showed similar changes in the fermentation pattern, with an increase in the molar percentage of valeric acid concentration in the colon. The SYN group, however, did not show any of the outcomes registered for PRO and PRE in Salmonella colonization or in immunity markers, suggesting the lack of synbiotic action in this animal model. Further research is needed to better understand the complex mechanisms behind these effects

Response to a Salmonella Typhimurium challenge in piglets supplemented with protected sodium butyrate or Bacillus licheniformis : effects on performance, intestinal health and behavior

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 (10 9 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 × 10 8 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

Dynamics of microbial cecal lumen ecosystem of broiler chickens with age measured by Terminal Restriction Fragment Length Polymorphism (t-RFLP)

Nowadays, it is known that intestinal microflora of broiler chickens plays an important role for growth performance and health. However, there is still a limited knowledge about the process of colonization of the chick gut