Diana Luise

Effect of feed supplementation with live yeast on the intestinal transcriptome profile of weaning pigs orally challenged with Escherichia coli F4

The ability of live yeasts to modulate pig intestinal cell signals in response to infection with Escherichia coli F4ac (ETEC) has not been studied in-depth. The aim of this trial was to evaluate the effect of Saccharomyces cerevisiae CNCM I-4407 (Sc), supplied at different times, on the transcriptome profile of the jejunal mucosa of pigs 24 h after infection with ETEC. In total, 20 piglets selected to be ETEC-susceptible were weaned at 24 days of age (day 0) and allotted by litter to one of following groups: control (CO), CO+colistin (AB), CO+5×1010 colony-forming unit (CFU) Sc/kg feed, from day 0 (PR) and CO+5×1010 CFU Sc/kg feed from day 7 (CM). On day 7, the pigs were orally challenged with ETEC and were slaughtered 24 h later after blood sampling for haptoglobin (Hp) and C-reactive protein (CRP) determination. The jejunal mucosa was sampled (1) for morphometry; (2) for quantification of proliferation, apoptosis and zonula occludens (ZO-1); (3) to carry out the microarray analysis. A functional analysis was carried out using Gene Set Enrichment Analysis. The normalized enrichment score (NES) was calculated for each gene set, and statistical significance was defined when the False Discovery Rate % was <25 and P-values of NES were <0.05. The blood concentration of CRP and Hp, and the score for ZO-1 integrity on the jejunal villi did not differ between groups. The intestinal crypts were deeper in the AB (P=0.05) and the yeast groups (P<0.05) than in the CO group. Antibiotic treatment increased the number of mitotic cells in intestinal villi as compared with the control group (P<0.05). The PR group tended to increase the mitotic cells in villi and crypts and tended to reduce the cells in apoptosis as compared with the CM group. The transcriptome profiles of the AB and PR groups were similar. In both groups, the gene sets involved in mitosis and in mitochondria development ranked the highest, whereas in the CO group, the gene sets related to cell junction and anion channels were affected. In the CM group, the gene sets linked to the metabolic process, and transcription ranked the highest; a gene set linked with a negative effect on growth was also affected. In conclusion, the constant supplementation in the feed with the strain of yeast tested was effective in counteracting the detrimental effect of ETEC infection in susceptible pigs limits the early activation of the gene sets related to the impairment of the jejunal mucosa.

Metabolomics characterization of colostrum in three sow breeds and its influences on piglets’ survival and litter growth rates

BackgroundColostrum is the first secretion produced by mammary glands during the hours immediately preceding and succeeding parturition. This secretion differs from milk and represents an essential vehicle of passive immunity, prebiotic compounds and growth factors involved in intestinal development. Most of the literature concerning colostrum composition refers mainly to human and cow; and little is known about pig colostrum metabolome and how it varies between pig breeds and different farrowing parity. Thus, the aim of the present research is to provide new information about pig colostrum composition and the associations between metabolites, the sows’ breed and the survival and growth rates of their litters.ResultsColostrum samples were gathered from 58 parturitions of sows belonging to three different breeds chosen for their importance in Italian heavy pig production: 31 Large White, 15 Landrace and 12 Duroc respectively. The defatted and ultrafiltered colostrum samples were analysed using 1H–NMR spectroscopy. Principal Components Analysis (PCA) was assessed on the obtained spectra. In addition, using a Stepwise Regression and a Linear Regression analyses the metabolites named after the signals assignment were tested for their associations with piglets’ performances. Twenty-five metabolites were identified, comprehending monosaccharides, disaccharides (such as lactose), organic acids (lactate, citrate, acetate and formate), nitrogenous organic acids (such as creatine) and other compounds, including nucleotides. PCA results evidence a clustering due to breed and season effects. Lactose was the main compound determining the assignment of the samples into different clusters according to the sow breed. Furthermore, some metabolites showed to be associated with piglets’ performance and survival traits: acetate and taurine were positively related to litter weight gain and piglets’ survival rate, respectively, while dimethylamine and cis-aconitate were linked to new-borns’ impaired ability to survive.ConclusionsThe results obtained suggest that colostrum composition is affected by breed, which, together with environmental conditions, may cause changes in colostrum metabolites content with possible consequences on piglets’ performances. Among the identified metabolites, acetate, taurine, dimethylamine and cis-aconitate showed consistent associations with piglets’ survival rate and litter weight gain, implying that these compounds may affect new-borns’ ability to survive.

The effects of starter microbiota and the early life feeding of medium chain triglycerides on the gastric transcriptome profile of 2- or 3-week-old cesarean delivered piglets

BackgroundThe stomach is an underestimated key interface between the ingesta and the digestive system, affecting the digestion and playing an important role in several endocrine functions. The quality of starter microbiota and the early life feeding of medium chain triglycerides may affect porcine gastric maturation. Two trials (T1, T2) were carried out on 12 and 24 cesarean-delivered piglets (birth, d0), divided over two microbiota treatments, but slaughtered and sampled at two or three weeks of age, respectively. All piglets were fed orally: sow serum (T1) or pasteurized sow colostrum (T2) on d0; simple starter microbiota (Lactobacillus amylovorus, Clostridium glycolicum and Parabacteroides spp.) (d1-d3); complex microbiota inoculum (sow diluted feces, CA) or a placebo (simple association, SA) (d3-d4) and milk replacer ad libitum (d0-d4). The The T1 piglets and half of the T2 piglets were then fed a moist diet (CTRL); the remaining half of the T2 piglets were fed the CTRL diet fortified with medium chain triglycerides and 7% coconut oil (MCT). Total mRNA from the oxyntic mucosa was analyzed using Affymetrix©Porcine Gene array strips. Exploratory functional analysis of the resulting values was carried out using Gene Set Enrichment Analysis.ResultsComplex microbiota upregulated 11 gene sets in piglets of each age group vs. SA. Of these sets, 6 were upregulated at both ages, including the set of gene markers of oxyntic mucosa. In comparison with the piglets receiving SA, the CA enriched the genes in the sets related to interferon response when the CTRL diet was given while the same sets were impoverished by CA with the MCT diet.ConclusionsEarly colonization with a complex starter microbiota promoted the functional maturation of the oxyntic mucosa in an age-dependent manner. The dietary fatty acid source may have affected the recruitment and the maturation of the immune cells, particularly when the piglets were early associated with a simplified starter microbiota.

Molecular networks affected by neonatal microbial colonization in porcine jejunum, luminally perfused with enterotoxigenic Escherichia coli, F4ac fimbria or Lactobacillus amylovorus

The development of an early complex gut microbiota may play an important role in the protection against intestinal dysbiosis later in life. The significance of the developed microbiota for gut barrier functionality upon interaction with pathogenic or beneficial bacteria is largely unknown. The transcriptome of differently perfused jejunal loops of 12 caesarian-derived pigs, neonatally associated with microbiota of different complexity, was studied. Piglets received pasteurized sow colostrum at birth (d0), a starter microbiota (Lactobacillus amylovorus (LAM), Clostridium glycolicum, and Parabacteroides) on d1-d3, and a placebo inoculant (simple association, SA) or an inoculant consisting of sow’s diluted feces (complex association, CA) on d3-d4. On d 26–37, jejunal loops were perfused for 8 h with either enterotoxigenic Escherichia coli F4 (ETEC), purified F4 fimbriae, LAM or saline control (CTRL). Gene expression of each intestinal loop was analyzed by Affymetrix Porcine Gene 1.1_ST array strips. Gene Set Enrichment Analysis was performed on expression values. Compared to CTRL, 184 and 74; 2 and 139; 2 and 48 gene sets, were up- and down-regulated by ETEC, F4 and LAM, respectively. ETEC up-regulated networks related to inflammatory and immune responses, RNA processing, and mitosis. There was a limited overlap in up-regulated gene sets between ETEC and F4 fimbriae. LAM down-regulated genes related to inflammatory and immune responses, as well as to cellular compound metabolism. In CA pigs, 57 gene sets were up-regulated by CA, while 73 were down-regulated compared to SA. CA up-regulated gene sets related to lymphocyte modulation and to cellular defense in all loop perfusions. In CA pigs, compared to SA pigs, genes for chemokine and cytokine activity and for response to external stimuli were down-regulated in ETEC-perfused loops and up-regulated in CTRL. The results highlight the importance of the nature of neonatal microbial colonization in the response to microbial stimuli later in life.

Effects of alpha-(1,2)-fucosyltransferase genotype variants on plasma metabolome, immune responses and gastrointestinal bacterial enumeration of pigs pre- and post-weaning

In pigs, the alpha-(1,2) fucosyltransferase (FUT1) gene has been highlighted for its properties in controlling the intestinal expression of enterotoxigenic E. coli (ETEC) F18 receptors; a pathogen causing edema disease and post-weaning diarrhoea. In this study, we hypothesized that pigs with different genotypes (ETEC F18 resistant (FUT1AA) versus susceptible (FUT1AG)) differed in following systemic and enteric responses: growth performance, plasma metabolic profiles, expression of candidate genes for intestinal mucosal homeostasis and immunity, number of selected bacteria and the concentration of short-chain fatty acids (SCFA) in faeces and digesta in piglets pre and post-weaning, and on the ETEC F18 adherence ex vivo. Genotype had the strongest impact on plasma metabolomic profile on day 7 and 28 of age. FUT1AG piglets had higher level of N-methyl-2-pyrrolidinone, hippuric acid, oxindole, and 3-oxo-5-beta-chol-7-en-24-oic acid on day 7, and a higher level of guanosine on day 28 than that in the FUT1AA piglets. FUT1AA piglets had a higher level of betaine on day 7 and 3-methylguanine on day 28. On day 34 of age, the FUT1AA pigs had higher levels of S-2-hydroxyglutarate, L-phenylalanine, tauroursodeoxycholic acid and an undetermined PC/LysoPC, while Ile Glu Phe Gly peptide and genistein 5-O-glucuronide, and PC (18:0/0:0) were at higher levels in the FUT1AG piglets. FUT1 genotype did not affect the growth performance and expression of candidate genes. FUT1AG piglets had a higher number of haemolytic bacteria in faeces and in digesta than that in FUT1AA at 34 days of age. The colonic acetic acid concentration was highest in FUT1AG piglets. FUT1 genotype may influence not only the expression of E. coli F18 receptors but could potentially impact the gut homeostasis and metabotype of piglets pre and post-weaning. Further investigations on the relation between FUT1 genotype and these aspects including the intestinal commensal microbiota will expand the knowledge on factors affecting the intestinal ecosystem.

Evaluation of Breed and Parity Order Effects on the Lipid Composition of Porcine Colostrum

Porcine colostrum lipid classes and fatty acids (FA) were characterized in 6 pools (from 69 samples) from 3 sow breeds (Italian Large White, Italian Landrace, and Italian Duroc) and different parity orders (only Large White). Triacylglycerols (TAG; 94.44 expressed as g/100 g of fat) were the most abundant lipid class, followed by diacylglycerols (DAG; 3.36 g/100 g of fat), free fatty acids (FFA; 0.98 g/100 g of fat), and cholesterol (0.84 g/100 g of fat). The main FAs found in swine colostrum were palmitic (27.29%, expressed as g/100 g of total FA), oleic (28.81%), and linoleic (23.39%) acids. Both the breed of sow and parity order affected the FA and lipid composition. The results suggest that the FA composition of swine colostrum is similar to that of human colostrum and could represent a new source of nutrients for human infants, after further assessment of hygienic and quality aspects. The swine model could be an opportunity for a better understanding of colostrum effects on newborns.

Genome wide association study for intramuscular fat in Italian Large White pigs using SNP60K chip

Whether cattle grazing in nature reserves in temperate summers ought to be provided with artificial shelter (man-made), in addition to natural shelter (vegetation), is a topic of debate. We have investigated the effect of heat-load on the use of natural versus artificial shelter (with a roof and three walls) by cattle in eight nature reserves in Belgium. GPS collars were used to monitor use of open area, natural and artificial shelter during one or two summers (per 30 min). Cattle location data were coupled to same-time values of climatic ‘heat-stress indices’ calculated from local weather stations’ measurements of air temperature, air humidity, solar radiation and wind speed. Use of open area decreased as heat-load increased. The strength of the effect, and whether the cattle sought natural or artificial shelter, were associated with the amount and spatial distribution of natural shelter in the reserve. When natural shelter was sparse, a more scattered distribution tempered the increased use of shelter with increasing heat-load. If sufficiently available, cattle preferred natural to artificial shelter. When little natural shelter was available, cattle did use the artificial shelter and especially so with increasing heat-load. Microclimatic measurements indicated that solar radiation was blocked by vegetation at least as well as by artificial shelter, and allowed more evaporative cooling. In conclusion, we found no evidence for the added value of additional artificial shelter to protect cattle from heat-load in temperate nature reserves, as long as adequate natural shelter is available.Trabajo presentado al 65th Annual Meeting of the European Federation of Animal Science (EAAP) (Copenhagen, Denmark, 25 al 28 de agosto, 2014).Trabajo presentado al 65th Annual Meeting of the European Federation of Animal Science (EAAP) (Copenhagen, Denmark, 25 al 28 de agosto, 2014).Intramuscular fat content (IMF) influences important qualitative traits of meat as tenderness, juiciness and flavour, and technological characteristics. This trait is difficult to measure in vivo and is not included in the breeding programs, despite its medium heritability. Furthermore, IMF is a complex quantitative trait determined by several biochemical and metabolic processes influencing fat deposition in muscles. Indeed QTL affecting this trait have been already reported and some candidate genes were investigated in the last years, but relevant causative mutations have not been so far detected. Recently, thanks to the sequencing of the porcine genome and to the development of a high throughput genotyping porcine chip, it is possible to perform genome wide association (GWA) studies and put in light markers associated to this trait. With the aim to identify genes and markers associated to IMF we performed GWA using the Illumina PorcineSNP60 BeadChip and 889 Italian Large White pigs included in the Sib Test genetic evaluation program of the Italian Association of Pig Breeders (ANAS). The association analyses were conducted using linear mixed model implemented in GenABEL. The results of GWA analysis allowed to detect seven markers significantly associated with IMF (P-value <10) mapped on chromosomes 1, 3, 8 and 12. A validation study on selected markers was performed using a mixed model procedure of SAS software. Markers identified, once confirmed, could be applied as candidate genes to improve meat quality traits in Italian Large White pigs.The constantly growing yearly demand for meat, dairy products and eggs has important implications for agricultural production methods. Nowadays livestock/crop production is becoming increasingly industrialised worldwide, shifting from extensive, small-scale, subsistence production systems towards more intensive, large-scale, geographicallyconcentrated, specialised and commercially oriented ones. The shift in livestock farming methods from extensive to intensive poses a number of significant challenges for animal welfare, environmental sustainability and food security. The indicators to assess animals health and welfare status, have been increased during the last years, and the importance of this discipline is now widely known worldwide. Thanks to the welfare quality ® protocol the procedure to assess the animal health and welfare has become more and more clear, precise and accurate since the project ended in 2009. Furthermore this procedure to assess animal welfare status is time consuming and requires manpower and accurate planning.Bearing in mind the objectives of this study to investigate the Barbary sheep fertility under our climatic conditions and to evaluate the efficiency of the prostaglandin f2α(pgf2α)injections in induction of fertile estrus in different seasons. For these objectives, some experiments were performed during the period from July,2008 to March, 2009. A total number of 300 Libyan Barbary ewes(3-6 years old, weighing 40-60 kg) was used in this experiment. Ewes were kept in privet farms, fed and managed similarly. Ewes were divided to four season groups(summer, autumn, winter and spring groups). Each season group was divided into treated and control. Animals in the treated groups were injected with double injections of 125 μg of prostaglandin f2α intramuscularly(i/m), 11 days apart. While those in control groups were injected with two injections of 1.0 ml of 0.9% NACL saline solution simultaneously with the treated ewes. At the same day (day 11) rams wearing painted sponges on their briskets regions were introduced for natural mating. Treated groups showed shorter estrus response time than control groups in all seasons(P<0.05). Estrus duration was longer in winter and spring than in summer and autumn (P<0.001),but no difference was found between treated and control groups inside seasons. Treated group showed higher pregnancy rate(P<0.001) in winter season than control group. Percentage of ewes lambed in winter was significantly high (P<0.001) among treated ewes than control (80 vs 38%). Lambing rate differed significantly (P<0.001) among treated groups in all seasons. Data were collected and calculated statistically using SPSS system for percentages, means, standard deviation(mean ± sd) analysis of variance (anova), Chi square and Dunacan’s test were used accordingly. Other values were measured, calculated and analyzed similarly. Adoption of technology in sheep farms of La Mancha, Spain J. Rivas1, C. De Pablos2, J. Perea1, C. Barba1, R. Dios-Palomares1, M. Morantes1 and A. García1 1Universidad de Córdoba, Campus Rabanales, 14014, Córdoba, Spain, 2Universidad Rey Juan Carlos, Paseo de los Artilleros, 28034, Madrid, Spain; pa1gamaa@uco.es Recently in Spain the milk production coming from sheep farms shows higher degrees of specialization based on the adoption of technologies. The aim of this research is to examine the pattern of adoption of technologies in sheep farms of La Mancha. Based on previous researches, from 77 questions, only 38 questions were selected by using qualitative and participatory methods; the chosen variables were grouped into six technology packages (TP): management, feeding, animal health and milk quality, pasture and land use, equipment and facilities, and reproduction and breeding program. The survey was applied to a sample of 157 farms. Using descriptive statistics each TP was characterized and the pattern of adoption was determined. The average of technologies adopted was of 18.4±6.0 (48.3%). TP showing higher degrees of implementation are animal health and milk quality (67.8%), feeding (56.0%) and management (55.7%), but their adoption is not sequential or responds to independent events. This research facilitates the identification of a number of technologies that must be implemented from an organizational strategy point of view. Moreover, all technologies are seeking a dynamic balance system that allows firms migrate to more efficient processes without losing their main attributes. As a technological challenge, an andrologic evaluation of ram, gynecologic evaluation of ewes prior to mating, early detection of non-productive animals are proposed; and a better use of productive records for the decision-making; aspects are recommended. The results of this analysis will have an impact on future research that attempts to improve the use of subproducts, forage reserves and improved rangeland management and hygiene control system, taking the quality milk as an strategic asset, so further research is necessary to assess the impact of each technology on the operating of the mixed system in the Mancha region. Session 40 Poster 20 Session 40 Poster 19The present paper focuses on evaluating the Interobserver Reliability of the Animal Welfare Assessment Protocol for Growing Pigs. The protocol for growing pigs consists of a Qualitative Behaviour Assessment (QBA), direct Behaviour Observations (BO), carried out by instantaneous scan sampling, a Human Animal Relationship Test (HAR) and checks for different Individual Parameters (IP), e.g. presence of tail biting, wounds and bursitis. Three trained observers collected the data by performing 29 combined assessments, which were done at the same time and on the same animals; but they were carried out completely independent of each other. The findings were compared by the calculation of Spearman Rank Correlation Coefficients (RS), Intraclass Correlation Coefficients (ICC), Smallest Detectable Changes (SDC) and Limits of Agreements (LoA). There was no agreement found concerning the adjectives belonging to the QBA (e.g. active: RS: 0.50, ICC: 0.30, SDC: 0.38, LoA: -0.05-0.45; fearful: RS: 0.06, ICC: 0.0, SDC: 0.26, LoA: -0.20-0.30). In contrast, the BO showed good agreement (e.g. social behaviour: RS: 0.45, ICC: 0.50, SDC: 0.09, LoA: -0.09-0.03 use of enrichment material: RS: 0.75, ICC: 0.68, SDC: 0.06, LoA: -0.03-0.03). The rather low agreement of the HAR (RS: 0.38, ICC: 0.54, SDC: 0.34, LoA: -0.40-0.27) can be explained by the fact that the observers entered the pens one after the other to minimise mutual interference which influenced the reaction towards the second intruder. Overall, observers agreed well in the IP, e.g. tail biting (RS: 0.52, ICC: 0.88; SDC: 0.05, LoA: -0.01-0.02) and wounds (RS: 0.43, ICC: 0.59, SDC: 0.10, LoA: -0.09-0.10). The parameter bursitis, however, showed great differences (RS: 0.10, ICC: 0.0, SDC: 0.35, LoA: -0.37-0.40), which can be explained by difficulties in the assessment when the animals moved around quickly or their legs were soiled. In conclusion, the Interobserver Reliability was good in the BO and most individual IP, but not for the parameter bursitis and the QBA.