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Dr. Barbara Metzler-Zebeli Mr. Andor Molnar Ms. Janine Scholz

Improving feed efficiency by understanding the intestinal bacterial network in pigs and poultry. Dr. Stefan G. Buzoianu Dr. Peadar G. Lawlor Ms. Ursula McCormack Moorepark Research Centre, Teagasc, Ireland. Dr. Barbara Metzler-Zebeli Mr. Andor Molnar Ms. Janine Scholz

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Dr. Barbara Metzler-Zebeli Mr. Andor Molnar Ms. Janine Scholz

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  1. Improving feed efficiency by understanding the intestinal bacterial network in pigs and poultry Dr. Stefan G. Buzoianu Dr. Peadar G. Lawlor Ms. Ursula McCormack Moorepark Research Centre, Teagasc, Ireland Dr. Barbara Metzler-Zebeli Mr. Andor Molnar Ms. Janine Scholz University ofVeterinaryMedicine Vienna

  2. Introduction

  3. ECO-FCE overview • Feb 2013 – Feb 2017 • 17 partners • 7 WPs • Overall objectives • improve food security by optimisingthe feed efficiency in pigs and broilers without negatively affecting animal welfare and meat quality • reduce the ecological footprint of the pig and broiler production systems • WP 3 objective • to examine the gut structure, function, microbiota and metagenomics in animals divergent for feed efficiency

  4. Work Package 3 Health & welfare Genomics Meat quality

  5. Determination offeedefficiency Selection of high and low feed efficient animals - based on Residual Feed Intake Residual Feed Intake (RFI) = difference between observed and predicted feed intake, with lower RFI values indicating greater energy efficiency RFI = FI [a + b1 * BW0.75 + b2 * BWG] Where a is the intercept and b1 and b2 are partial regression coefficients of feed intake (FI) on BW0.75 and body weight gain (BWG), respectively. Other measures of feed efficiency Feed efficiency = gain (g) / feed intake (g) Feed conversion ratio (FCR) = feed intake (g) / gain (g) RG = BWG [a + b1 * BW0.75 + b2 * FI] RIG = (RG/SD RG) - (RFI/SD RFI)

  6. Feed efficiency in monogastric livestock species Gut commensalmicrobiota Substantial variation in feedefficiencybetween individual animals. Great variation in gut commensalmicrobiotabetween individuals.

  7. Role of the intestinal microbiota • Costs to the host • competition for nutrients • immune activation • production of toxins • opportunistic • toxin reabsorption • mucolytic activity • Benefits to the host • intestinal maturation • inhibition of pathogen growth • nutrient salvaging • detoxification • production of vitamins

  8. Effectofhostmicrobiota on hostmetabolismandhormonesecretion Intestinal microbiotacanredirectenergypartitioningtoadiposetissueandreducefattyacidoxidation. • Implicationsforfeeduseefficiencyandcarcasscomposition in livestock animals? Bäckhed (2011) Ann NutrMetab 58(suppl 2):44

  9. Effectof gut microbiotacomposition on bodyweight Obesehumans & mice: Firmicutes Bacteroidetes  Low-caloriediet Firmicutes Bacteroidetes  wikipedia.org Actinobacteria Bacteroidetes  nodifference in Firmicutes Changes in LactobacillusandBifidobacteriumspecies Methanogenicarchaea Requena et al. (2013) Trends Food Sci Tech 34:44 Meat-producingmonogastric livestock speciesareyoung, fast growingandleananimals Are thekeyplayersthe same as in human obesitymodels ?

  10. Chickens

  11. Diet-relatedcecalmicrobiotaandperformance in male chickens Caecalmicrobialcommunitiesbydiet Caecalmicrobialcommunitiesidentifiedasbeingfrombirdswithimprovedperformanceorpoorerperformance Dietisthemostinfluencingfactoraffectingfeedefficiency. Torok et al. (2011) AEM 77: 5868

  12. Batch tobatchvariation in caecalmicrobiotaofchickens 3 different batchesofchickens PCA plotofcaecalmicrobiota. The plotisbased on betweengroups (trials) analysis. Very different microbiotaprofilesacrosschickenbatches Very different feed use efficiencies across chicken batches Stanley et al. (2013) PloS ONE 8(12): e84290 • High variation in caecalmicrobiotapartly due to lack ofcolonisationofthechickensbymaternallyderivedbacteria • High hygienelevels in modern commercialhatcheriesremovenaturalbacteria • Environmental microbiotafromtransportboxes, firstfeedandstaffpeople

  13. Fecalcommunityofhighandlowfeedefficientbroilerchickens Singh et al. (2014) J Appl Genet 55: 145

  14. Characterisationofdifferences in gut microbiotaand gut functionofchickenswithgoodandpoorfeedefficiency • Experimental design: • 2 partnerinstitutions (AFBI & Vetmeduni) performedidenticalchickenexperimentswith 3 batchesof 50/64 chicks • Similarchickengenetic: Cobb 500FF • Similarmaize-soybeanmealdiets (starter, grower, andfinisherdiets) • No in-feedantibioticsandanyother gut health-related additives • Chickenswere individually housed • Best andworstfeedefficientchickenswereidentifiedusing Residual Feed Intake • On day 42, sampleswerecollectedfor: • Ilealandcaecaldigestaformetagenomicsandmicrobialmetabolites • Tissueofduodenum, jejunum, ileum, caecafor gut functionandstructure

  15. Residual feedintakeofgoodandpoorfeedefficientbroilerchickens Great variation in residual feedintakeandthus in feeduseefficiency.

  16. Microbialmetagenomeofgoodandpoorfeedefficientchickens Underconstruction • ShotgunsequencingusingMiSeq Technology (Illumina)

  17. Jejunal electrophysiologicalcharacteristicsofgoodandpoorfeedefficientbroilerchickens • Gut electrophysiology was performedusingUssingchambertechnique. • Tissueoriginatedfromthe distal jejunum. Good feed efficient females showed lower tissue resistance, higher conductance and short-circuit current indicating a higher ion flux and permeability of the jejunal mucosa Influencingfactors: Host genomeor gut microbiota ?

  18. Pigs

  19. Literature • Little data available in pigs • ↓ Bacteroidetes & ↑ Firmicutes in obese pigs (Pedersen et al., 2013) • ↑ Firmicutes & ↓ β-Proteobacteriain ERS-fed pigs (Haenenet al., 2013) • Protein, CHO and lipid metabolic pathways affected by intestinal microbial profile • mice (Antuneset al., 2011) • pigs (Mulder et al., 2009)

  20. Screening on feed efficiency in pigs P P d 42 d 84 weaning d 112 F – faecal I – ileal digesta C – caecal digesta P – performance F F F I C

  21. Microbiota profiling d 0 (weaning) d 42 d 84 d 126 d 139 P P P P F F F F F I C Compositional analysis 16S rRNA gene sequencing Functionality Shotgun metagenomics Illumina F – faecal; I – ileal digesta; C – caecal digesta; P – performance

  22. Progress on microbiota profiling • Samples collected • DNA extracted • 16S rRNA gene sequencing – results being analysed • Shotgun metagenomics • samples being prepared • results ~ Oct 2014

  23. Low RFI Manipulation of GIT microbial profile Additives

  24. Inoculation with faecal inoculum from good feed converters • Anaerobically processed • diluted 1:6 • strained • centrifuged (6000 × G for 15 minutes) • frozen at -80°C in 10% glycerol Sows Offspring

  25. Nutritional intervention • Optimum strategy – inoculum • Prebiotics – alone or in combination • Monitoring and sampling of offspring through their lifetime • performance • health • intestinal microbiota

  26. Acknowledgements • ECO-FCE has received funding from the European Union’s Seventh Framework Programme for research, technological development and demonstration (FP7 2007/2013) under grant agreement No. 311794 • Teagasc Walsh Fellowship Programme

  27. Thank you

  28. ECO-FCE Gut structure, function, microbiotaandmetagenomics Objectives: • Toenhanceourunderstandingofthe interactions between gut microbiomeandhostgenome in pigsandchickens. Thistask will beachievedbyemployingcutting-edge 16S rRNA-specificandshotgunmetagenomics. • Usingthisimprovedunderstanding, strategies toimprovefeedconversionefficiencythrough gut microbiomemanipulation in embryonicand subsequent developmentalstages will bedeveloped. Hypothesis: Itisassumedthatthe gut microbiomeofpigsandbroilerchickenswithgoodandpoorfeeduseefficiencydiffers in keymembers, therebyinfluencingthe intestinal andmetabolichostresponse, productionefficiencyandhosthealth.

  29. Interactions between gut microbiomeandhostphysiologyandhealth Mucussecretion Barrierfunction Mucosalimmunity NF-kB Commensal microbiota Growth & feedefficiency Gut morphology Nutrientdigestibility Nutrient transporters Bäckhed (2011) Ann NutrMetab 58(suppl 2): 44; Twarziok et al. (2014) Mol Inf 33: 171

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