2017 PSA Annual Meeting

Oral Presentation

Effect of different levels of sorghum inclusion and the addition of a serine protease on live performance of broilers from 1 to 35 days of age.

Albaraa Sarsour*1, Hernan Cordova-Noboa1 , Edgar Oviedo-Rondón1 , Pedro Ferzola1 , Nasser Odetallah2 , 1North Carolina State University, Raleigh, NC, 2Novus International, St. Charles, MO.

Sorghum can be utilized to replace corn in areas where corn is not readily available. Serine protease has been proposed to improve the digestibility of sorghum because it could degrade disulfide bonds of kafirin proteins. One experiment was conducted to evaluate the effects of sorghum inclusion level and the addition of a serine protease on chicken live performance from 1 to 35 d of age. Eight treatments from a 4x2 factorial arrangement with 4 inclusion levels of sorghum (0, 25, 50, and 100% replacement of corn) and presence or absence of protease (0 or 500 g/ton for 600,000 U/g), as main factors. A total of 1,280 Ross 708 d-old male chicks were placed in 64 pens with 20 chicks per pen raised on used litter. Group BW and feed intake (FI) were recorded at 0, 14, and 35 d. BW gain and FCR corrected by mortality BW weights were calculated at the end of each phase. Data were analyzed as a randomized complete block design with a 4x2 factorial arrangement with 8 replicates per treatment combination. There was an effect of sorghum level (P < 0.05) on BW at 35 d. The heaviest broilers were associated with the treatments that were fed 100% corn diets which were not different from 25 and 50% sorghum, but were heavier (P < 0.05) than the ones fed 100% sorghum diets. Similar effect (P < 0.05) was observed on BW gain from 14 to 35 d, and BW and FI from 1 to 35 d. There was a 2-way interaction effect (P < 0.05) observed on FI for sorghum level and enzyme. Broilers fed diets with 100% sorghum without the addition of a protease consumed less (P < 0.05) feed than the same diet with the addition of protease. Protease improved FCR in the starter from 1 to 14 d only in diets with 100% corn. This positive effect of protease (P < 0.05) on FCR was detected again from 14 to 35 and 1 to 35 d of age in diets with 100% corn. It was concluded that corn can be replaced with up to 50% sorghum and that the addition of a protease could improve FCR of broilers fed corn diets and FI in 100% sorghum diets.

Key Words: sorghum level, serine protease, broiler performance

Poster Presentation

Effect of chelated copper on growth performance and woody breast in broilers J. Chen, F. Yan, V. Kuttappan, and M. Vazquez Anon Novus International Inc., St. Charles, MO, USA

Copper (Cu) has been widely used at high levels as growth promoter in poultry. The role of Cu in woody breast has not been widely studied. There are different forms of chelated and inorganic Cu sources available for the poultry producers to use at different doses. A floor pen study was conducted with 468 day-old male broilers to evaluate the effects of Cu methionine hydroxyanalogue chelate (Cu-MHAC) (MINTREX® Cu, Novus International, Inc.) on growth performance and incidence of woody breast in broilers in comparison to CuSO4 and TBCC. The study consisted of 4 dietary treatments: 15 ppm Cu-MHAC, 30 ppm Cu-MHAC, 125 ppm CuSO4 and 125 ppm TBCC, each with 9 replicates pens of 13 birds. The levels of other minerals from inorganic sources were equal among all treatments. Nutritionally complete typical US corn soybean meal based broiler diets were formulated for starter (0-14 d), grower (15-28 d), and finisher (29-42 d) phases. All diets were pelleted, and starter diet was crumbled after pelleting. All birds were orally gavaged with a coccidiosis vaccine at 10× the recommended vaccination dose on d 15. Breast fillets in broilers at 42 d of age were scored for woody breast using 4 points scoring system: 0 = normal; 1 = mild; 2 = moderate; 3 = severe. Woody breast results were analyzed by Chi-square. Performance results were analyzed by one-way ANOVA, means were separated by Fisher’s protected LSD test. A P-Value ≤ 0.05 was considered statistically different. Both 15 and 30 ppm Cu-MHAC improved (P<0.05) FCR in comparison to CuSO4, and TBCC had intermediate FCR during starter phase. Growth performance was not different among treatments during grower and finisher phases. 15 ppm Cu-MHAC reduced (P<0.05) the incidence of moderate and severe woody breast compared to TBCC; while 30 ppm Cu-MHAC reduced (P<0.05) the incidence of moderate and severe woody breast compared to both CuSO4 and TBCC. In summary, low doses of Mintrex Cu can replace high dose of TBCC and CuSO4 without compromising growth performance and with benefits of reducing the incidence of moderate and severe woody breast in broilers.

Keywords: Chelated copper, CuSO4, TBCC, woody breast, broiler

Poster Presentation

Estimation of true phosphorus digestibility of soybean meal is affected by Ca level in broilers.

Colwayne Morris*2, Roger Davin1 , Fenglan Yan1 , Megharaja Manangi1 , David Ledoux2 , Mercedes Vazquez1 , 1Novus International Inc, Saint Charles, MO, 2University of Missouri, Columbia, MO.

Previous regression analysis estimations of true ileal P digestibility (TIPD) in SBM using a fixed Ca:total P ratio predicted values lower than apparent ileal digestibility (AIPD) values in broilers. The objective of the current study was to estimate TIPD in SBM by regression analyses, using 2 fixed Ca levels (0.35 and 0.85%), and to estimate endogenous P losses (EPL) by regression analysis and by providing P-free diets. A total of 320 Ross 308 male broiler chicks were fed a common corn-SBM based diet from d 1 to d 19. From d 19 to 22, birds were fed 8 experimental diets that consisted of 3 graded levels (20, 40 and 60%; low, medium and high, respectively) of SBM and 2 gelatin-based P-free diets containing either 0.35% or 0.85% Ca. Each treatment was fed to 8 replicate cages with 5 birds/cage. Digesta from the posterior 2/3rd of the ileum was collected on d 22. Data were analyzed by one-way ANOVA to determine statistical differences among treatments. TIPD and endogenous P loss estimates were obtained by regressing P output against dietary P content in SBM diets. TIPD of SBM with 0.85% Ca was estimated to be 59.3%, a value 19 points lower than the TIPD estimation of SBM with 0.35% Ca (78.3%). TIPD estimate values were greater than the AIPD values obtained from the SBM diets supplemented at different Ca inclusion levels. Increasing SBM level from low to medium improved AIPD at both Ca concentrations. However, increasing SBM from 40 to 60% did not further increase AIPD. As previously reported, EPL estimates were significantly affected by the methodology employed, being 341 and 48.9 mg/kg DMI using the regression approach and P-free diets, respectively. Ca level did not affect EPL (P > 0.4) in both methodologies. In summary, TIPD values from SBM were greater than AIPD values when a fixed dietary Ca level is employed, however Ca concentration has a significant impact on both TIPD and AIPD. EPL estimates vary depending on the methodology employed.

Key Words: digestibility, endogenous losses, phosphorus, soybean meal

Poster Presentation

Evaluation of Eimeria-dietary challenge model to test the benefits of alternatives for antibiotic growth promoters in broiler birds.

Vivek Kuttappan*1, Juxing Chen1 , Fenglan Yan1 , Mercedes Vazquez-Anon1 , 1Novus International Inc, St. Charles, MO.

Eimeria infection is a major reason for economic loss to the modern poultry industry due to its direct impact on performance, and being a major predisposing factor for dysbacteriosis leading to necrotic enteritis. The main objective of present study was to develop an Eimeria-dietary challenge model to test the efficiency of various alternatives for antibiotic growth promoters (AGP). In experiment 1, doses of 0, 20, 50, and 100× of a commercial coccidiosis vaccine containing 3 different species of Eimeria were tested in birds fed with either corn-soy (DIET-1) or corn-soy with DDGS/animal protein (DIET-2) diets (8 replicates/treatment; 8 birds/replicate) throughout the trial. Birds were challenged with Eimeria on 14d of age, and performance as well as serum color were measured on 23d of age. In experiment 2, the treatment groups (10 replicates/treatment; 10 birds/replicate) included no challenge control (NC), challenged/positive control (PC; challenged with 20× commercial vaccine on 14d), PC + Probiotic (spore based probiotic), PC + Essential oil (NEXT ENHANCE 150; NE150), and PC + bacitracin methylene disalicylate (BMD). All the birds were fed with DIET-2 throughout the trial. On 23d performance as well as serum parameters were measured. Data were analyzed using ANOVA and means were separated with Fisher’s protected LSD test at P < 0.05. Results from experiment 1 showed that as the dose of Eimeria challenge increased there was a reduction (P < 0.05) in body weight, cumulative feed conversion (cFCR), cumulative performance index (cPI), and serum color (an indicator of nutrient absorption). Furthermore, birds fed with DIET-2 had lower (P < 0.05) cFCR and cPI than DIET- 1. In Experiment 2, PC showed reduction (P < 0.05) in body weight, cFCR, cPI, and serum color compared with NC. In addition, Eimeria challenge also resulted in increased (P < 0.05) level of serum IL-10, an anti-inflammatory cytokine, in PC compared with NC. Interestingly, probiotic, NE150, and BMD showed improvement (P < 0.05) in cFCR and reduction in serum IL-10 levels when compared with PC birds. Therefore, BMD and AGP alternatives such as probiotics and essential oils can improve performance in broiler birds under Eimeria challenge, plausibly by reducing tissue damage associated with the infection. In conclusion, Eimeria challenge along with corn-soy DDGS/animal protein diet could be an effective challenge model to test alternatives for AGPs such as BMD.

Key Words: Eimeria challenge, dietary challenge, probiotics, essential oil, antibiotic growth promoter

Poster Presentation

Performance benefits of probiotic and protease in broilers subject to Eimeria challenge

F. Yan, J. Chen, V. Kuttappan, and M. Vazquez Anon Novus International Inc. St. Charles, MO

Consumer pressure and government regulations are driving US poultry industry to remove antibiotic growth promoter (AGP) from poultry production. Nutrition program including feed additives is an integral part of AGP free poultry production. A floor pen study was conducted with 1152 day-old male broilers to evaluate the efficacy of probiotic and protease in alleviating the negative impact of mild Eimeria challenge on growth performance and gut health in comparison to Bacitracin Methylene Disalicylate (BMD®). The study consisted of 4 dietary treatments – Negative Control (NC), NC + probiotic (SPORULIN® containing spores of B. Subtilis), NC + protease (CIBENZA® DP100), and NC + BMD, each with 12 replicate pens of 24 birds. Nutritionally complete typical US corn soybean meal based diets were formulated for starter (0-14 d), grower (14- 27 d), and finisher (27-41 d) phases. All birds were challenged with a coccidiosis vaccine at 10X recommended dose via oral gavage on d 15. Body weight, feed intake, FCR, and mortality were determined on d 14, 21, 27, and 41. On d 28, 3 birds per pen were euthanized to score E. acervulina, E. maxima, and necrotic enteritis (NE) lesions. Data were subject to one way ANOVA; means were separated by Fisher’s protected LSD test. Bacitracin improved (P<0.05) FCR on d 14 (1.144 vs 1.220), d 21 (1.294 vs 1.352), and d 27 (1.375 vs 1.426); probiotic and protease improved (P<0.05) FCR on d 14 (1.150 and 1.153 vs 1.220) and d 21 (1.316 and 1.315 vs 1.352), and their effects were not different from BMD (P>0.05). No significant difference in FCR was observed among dietary treatments on d 41 (P>0.05). Bacitracin increased (P<0.05) BW by 6.0% and feed intake by 4.8% on d 21; probiotic and protease numerically increased (P>0.05) 21-d BW by 3.0 and 2.8% and feed intake by 2.2 and 2.8% to a level that was equivalent to BMD (P>0.05). No treatment difference in BW or feed intake was observed on d 14, 27 and 41 (P>0.05). Low mortality throughout the trial, minimal coccidiosis and mild NE lesions were observed, and they were not affected by dietary treatments (P>0.05). In summary, for broilers subject to Eimeria challenge on d 15, while BMD was effective in improving performance up to d 27, probiotic and protease were effective up to d 21 indicating that they have different mechanism of action from BMD, and they could serve as effective nutritional tools to maintain growth performance during certain phases in AGP free broiler production.

Keywords: probiotic, protease, antibiotic, broiler, Eimeria

Poster Presentation

The effect of combining microbial phytase, protease, and xylanase on performance of broiler chicks fed diets containing reduced levels of available phosphorus, amino acids and energy.

M. K. Manangi*, S. Bettis, J. Chen, and M. Vazquez-Anon; Novus International, Inc., St. Charles, MO.

A 42d experiment was conducted to evaluate the effects of supplementation of microbial 6- phytase (CIBENZA® PHYTAVERSE® G10) in combination with a protease (CIBENZA® DP100) and xylanase (CIBENZA® XYLAVERSE®) in broiler chicks fed reduced levels of available phosphorus (aP), amino acids (aa), and energy corresponding to matrix values assigned for each of the products using Corn-SBM-Canola meal-Rice bran mixed diets. A total of 2016 Ross-308 male chicks were assigned under randomized complete block design to 9 treatments with 8 pens/treatment and 28 chicks/pen. Treatments consisted of reduced levels of aP, aa/CP and energy from the positive control (PC) diet. Test diets included: T1 (negative control, NC); T2 (T1+phytase at 500U/kg diet); T3 (T1+protease at 300,000U/kg diet); T4 (T1+xylanase at 250U/kg diet); T5 (T1+phytase + protease); T6 (T1+phytase + xylanase); T7 (T1+ protease + xylanase); T8 (T1+phytase + protease + xylanase); T9 (industry levels of aP, aa/CP, and energy) (PC). For T1 the energy was reduced by ~90kcals, CP by ~5%, Lys by ~4%, TSAA by ~5%, and Thr by ~7%, across phases compared to T9. The data were analyzed using 1-way ANOVA and the means were separated using protected t test at P≤0.05. End of the trial outcome indicated a significant treatment effect for cumulative FCR (adjusted for mortality & culls) (cFCR) (P<0.05) but not for cumulative weight gain, feed intake, and mortality (P>0.05). cFCR for the PC was better (P<0.05) than all the treatments except T5. cFCR for T2, T4, T6, T7, and T8 were similar and not different from the NC (T1) (P>0.05). T3 (protease) improved (P<0.05) cFCR compared to the NC but not different from T5. Supplementing the combination of phytase and protease (T5) resulted in a significant improvement (P<0.05) in cFCR compared to T1 and similar (P>0.05) to the PC (T9). In summary, under the experimental conditions tested, the data from day 42 indicates that the combination of phytase and protease leads to improved cFCR which is similar to the PC than the enzymes supplemented separately or in other combinations. ®CIBENZA, PHYTAVERSE, and XYLAVERSE are trademarks of Novus International, Inc. and are registered in the United States and/or other countries.

Keywords: Phytase, Protease, Xylanase, Broiler

Oral Presentation

True P digestibility of corn, SBM and corn-soybean meal without and with phytase in broilers

Roger Davin*1, Colwayne Morris2 , Fenglan Yan1 , Megharaja K. Manangi1 , David Ledoux2 , Mercedes Vazquez1 ; 1Novus International Inc, Saint Charles, MO, 2University of Missouri, Columbia, MO.

The objective of the current study was to evaluate the true ileal P digestibility (TIPD) of corn, SBM, and corn soy based diet (C-SBM) as affected by phytase supplementation using a regression approach in broilers. A total of 800 Ross 308 male broiler chicks were fed a complete common corn-SBM-based diet from d 1 to d 15. From d 15 to 23, birds were fed 18 experimental diets. Three inclusion rates were provided for corn (22, 46 and 72%), SBM (40, 51 and 62%) and CSBM (21/10, 42/21 and 63/31% of a combination of corn/SBM) supplemented without or with 500 U of phytase/kg diet (CIBENZA® PHYTAVERSE®, Novus International, Inc., St Charles, MO). Corn diets contained 0.7% Na-phosphate and all diets had a Ca:P ratio between 1.3-1.5:1 following WPSA (2013) guidelines. C-SBM diets contained 0.91% dicalcium phosphate to mimic commercial diets. Each treatment had 8 replicate cages (5 birds/cage). Digesta from the posterior 2/3rd of the ileum was collected on d 23. Data were analyzed by 2-way ANOVA to evaluate the effect of ingredient level, phytase and their interaction, and by orthogonal polynomial contrasts to test the linear and quadratic effects of ingredient level. TIPD estimates were obtained by regressing P output against dietary P content. TIPD estimate of SBM (52.0%) is more reasonable than the extremely low values obtained for corn (-7.2%) and C-SBM (5.6%). Corn and C-SBM contained inorganic P, whereas SBM diets contained no inorganic P. Phytase improved TIPD in corn (+38 points, to 60.4%; P<0.01) and C-SBM (+14 points, to 19.5%; P=0.02), however it did not in SBM (+7 points, to 59.9%; P=0.55). Apparent ileal P digestibility (AIPD) tended to decrease linearly (P<0.08) as corn and SBM levels increased, and increased when phytase was supplemented. A significant ingredient level × phytase interaction was found for AIPD in corn-SBM (P<0.01). Phytase improved AIPD at all ingredient levels, however the response of ingredient level in diets without phytase (low, med, high: 76.2b, 54.7d, 45.7e%) was quadratic and linear for diets with phytase (low, med, high: 88.1a, 76.7b, 60.0c%). In summary, TIPD estimates for SBM and for corn and C-SBM obtained by regression are lower than AIPD and far from reality possibly due to the use of fixed Ca:P ratio and of inorganic P in corn and C-SBM diets. Phytase efficiently improved TIPD and AIPD in corn and C-SBM, and AIPD in SBM.

Keywords: Corn, Soybean meal, phytase, digestibility, phosphorus.

Oral Presentation

Genomic traits of a strain of Salmonella Heidelberg isolated in broilers in Brazil and related phenotypic tolerance to organic acids and antibiotics.

Elizabeth Santin*1, Ricardo Hayashi1 , Mariana Camargo Lorenco1 , Raquel Bighetti Araujo2 , Ricardo Gonzalez-Esquerra2 , Marcelo Falsarella Carazzolle3 , Cáio César de Melo Freire4 , Paulo Sergio Monzani5 , Anderson Ferreira da Cunha4 , 1Universidade Federal do Paraná, Curitiba, Parana, Brazil, 2Novus International Inc, Indaiatuba, SP, Brazil, 3Universidade Estadual de Campinas, Campinas, SP, Brazil, 4Universidade Federal de São Carlos, São Carlos, SP, Brazil, 5Universidade de São Paulo, Pirassununga, SP, Brazil.

Salmonella enterica serovar Heidelberg (SH) is found in broilers worldwide with isolates from Brazil (SHBR) increased since 2011 and showing greater tolerance to control measures. A previous trial showed that a probiotic composed of 3 Bacillus subtilis strains was effective vs. SHBR in broilers. Herein, we aimed at sequencing the genome of SHBR and relate genomic differences with SHBR tolerance to some organic acids (OA), antibiotics (AB); and clinical signs in broilers. Two trials used 1d old chicks housed for 21d in 8 sterilized isolated negative pressure rooms with 4 battery cages (reps) of 12 birds each. In both trials, birds were challenged or not with 107 cfu/bird of SHBR orally, and exposed, or not, to OA in a factorial 2x2 design. Challenge to SHBR occurred at 1 or 7d of age; and OA tested consisted of either formic + propionic acids in drinking water at 0.05% from 1 to 7d and 15–21d; or calcium butyrate fed at 2kg/ton of feed from 1 to 21d in trial 1 or 2, respectively. Nine AB were titrated in an in vitro MIC model using MuellerHinton agar as in CLSI[PM1] /NCCLS guidelines, to test SHBR-AB tolerance. SHBR DNA was sent to the High Throughput Sequencing Facility (University of North Carolina). The library was prepared using PacBio 20Kb template prep protocol PN_100–286–000–06, a size-selected range of 8000bp - 50,000bp, and the PacBio native pipeline for De novo assembly. The genome was deposited at NCBI genome database (No. CP020101) and compared with SH SL476 strain. Performance and immune response traits were unaffected by SHBR (P > 0.05). The use of OA did not reduce Salmonella counts found in cecum and liver of challenged birds (P > 0.05). SHBR was susceptible to amoxicillin-clavulanic acid, cephalosporin, ciprofloxacin, enrofloxacin, penicilin and trimethoprim-sulfamethoxazole and tetracyclin with mild resistance to gentamycin and ceftiofur. Several DNA fragments were missing in the SHBR genome which were associated with the codification of proteins involved with cell cycle regulation, virulence, drug resistance, cell adhesion, salt efflux, and various transposases and integrases that may relate to those deletions. These genomic findings relate to the phenotypic observations of low pathogenicity, OA tolerance and AB susceptibility of SHBR.

Key Words: Brazilian Salmonella Heidelberg, antibiotics, organic acids, resistance, comparative genomics