ID: 707

IS THE BROILER RESPONSE TO DIETARY VALINE MODULATED BY THE SAME BRANCHED-CHAIN AMINO ACIDS AS IN PIGLETS?

M. Zouaoui\*(1), A.Simongiovanni(2), W.Lambert(2), M.Létourneau montminy(1)

Branched-chain amino acids (BCAA), that include valine (Val), leucine (Leu) and isoleucine (Ile), share unique catabolism pathway, leading to strong metabolic interactions. Excess of dietary Leu was first shown in piglets to stimulate the degradation of all BCAA including Val and Ile, with negative consequences on feed intake and growth. The aim of this study was thus to quantify and compare the response of broilers and piglets to dietary Val and the impact of others BCAA on this response thanks to a meta-analysis approach. The databases were composed of 23 articles published between 1999 and 2018 including 44 experiments and 239 treatments for broilers, and 16 articles published between 2001 and 2018 including 23 experiments and 126 treatments for piglets. Multiple regression models were fitted with the MIXED procedure of Minitab software with experiment as random effect. The Y variables were average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR). For broilers, the Y variables were expressed in percentage of the highest-level intra-experiment to make the values comparable between experiments (different ages and genetic lines). The main X variable was the dietary standardised ileal digestible Val level (ValSID) and the other ones were dietary SID Leu and Ile (LeuSID and IleSID, respectively). The response of ADG and ADFI to ValSID was linear and quadratic for broilers and piglets (Broilers, ADG: P < 0.001, R2=0.65, ADFI: P < 0.001, R2=0.50; Piglets, ADG: P < 0.001, R2=0.93, ADFI: P < 0.001, R2=0.97). Results showed that increasing dietary LeuSID reduced ADG and ADFI in broilers and piglets (P < 0.05) but also that the response to ValSID was stronger with high LeuSID levels for these 2 species (Interaction ValSID×LeuSID: Broilers, ADG and ADFI: P < 0.001; Piglets, ADG and ADFI: P < 0.05). The response of ADG and ADFI to ValSID was not modified by IleSID. The FCR responded linearly (P < 0.001) and quadratically (P < 0.001) to ValSID, while LeuSID and IleSID have no effect on this response for both species. In conclusion, the current study showed that ADG and ADFI but not FCR response to ValSID is modulated by dietary LeuSID level. These results indicate that Leu is the most important regulator of BCAA metabolism that works similarly in broilers and pigs. This quantification will facilitate the decision-making on amino acid formulation constraints and the implementation of low protein diets for these two species.

<footnote>(1)Laval University, Quebec, Canada</footnote>

<footnote>(2) Ajinomoto Animal Nutrition Europe, Paris, France</footnote>

<footnote>* Corresponding author: marie-pierre.letourneau@fsaa.ulaval.ca</footnote>
EFFECTS OF L-ARGININE AND L-CITRULLINE SUPPLEMENTATION IN BROILERS FED REDUCED PROTEIN DIETS UNDER CYCLIC WARM TEMPERATURE

H. Dao*(1), N.Sharma(2), E.Bradbury(3), R.Swick(2)

(1)University of New England, Australia, Armidale, Australia

(2)University of New England, Armidale, Australia

(3)Ridley AgriProducts, Melbourne, Australia

* Corresponding author: tdao@myune.edu.au

This study aimed to determine the effects of L-arginine (Arg) and L-citrulline (Cit) supplementation on performance, carcass traits and gut permeability of broilers fed reduced protein diets and housed at thermo-neutral or cyclic warm temperature. One-day-old Ross 308 male birds (n = 720) were fed a common starter diet until d 7, and treatment diets during grower (d 8 to 21) and finisher (d 22 to 35) periods. On d 8, birds were weighed and randomly allocated to one of four dietary treatments with 12 replicates of 15 birds per pen. Dietary treatments were: normal protein (NP), reduced protein (RP) deficient in Arg, RP supplemented with Arg, and RP supplemented with Cit. Equivalent levels of Arg and Cit were added in grower and finisher diets at 0.29% and 0.28%, respectively. The difference in crude protein levels between NP and RP diets was approximately 25 g/kg. From day 22 to 35, the temperature was cycled up to 33°C ± 1°C for 6 hours per day in one room while the other room was continuously set at 240°C. On d 28, two birds per pen were selected, marked and inoculated with 1ml dose of Fluorescein isothiocyanate dextran (FITC-d), a leaky gut marker, to measure the gut permeability. Data from d 22 to 35 were analysed to show the main effect of temperature, treatment and their interaction. Dietary treatment x temperature effect was not observed for performance variables (P > 0.05). Regardless of temperature, birds fed Arg-deficient RP diets showed reduced weight gain and 2.3 points higher FCR compared to those fed NP diets (P < 0.05). Feeding Arg-deficient RP diets resulted in lower thigh and drumstick, and higher fat pad yields at d 35 compared to NP diets (P < 0.05). Supplementation of Arg in RP diet improved body weight gain and feed efficiency whereas Cit only improved feed efficiency (P < 0.05). Reduced protein diets supplemented with Arg or Cit improved carcass traits which were similar to those fed NP diets (P < 0.05). Regardless of the dietary treatment, birds exposed to the cyclic warm temperature showed reduced feed intake, body weight gain, and impaired gut permeability (higher serum FITC-d level) compared to those housed at thermo-neutral temperature (P < 0.05). In conclusion, broilers perform poorly in Arg deficient reduced protein diet and the supplementation of Arg or Cit in reduced protein diet helps to improve the performance of broilers. Further research on the effect of Arg and Cit during heat stress (temperature > 35°C) is warranted.
VALINE REQUIREMENT OF FEMALE BROILERS IN STARTER PHASE

S. Amirdahri(1), H.Janmohammadi*(1), W.Lambert(2), E.Asadi soumeh(3)

(1)university of Tabriz, Tabriz, Iran

(2)Ajinomoto Animal Nutrition Europe, Paris, France

(3)University of Queensland, Queensland, Australia

* Corresponding author: Janmohammadi@tabrizu.ac.ir

Valine is the 4th limiting amino acid in growing chicks, and deficient of the amino acid in diet, impair
growth performance. So, the purpose of this study was estimate the digestible valine to lysine ratio in
practical diets of starter female Cobb-500 broiler chicks. A total of 540 female broiler chickens of the
Cobb-500 from 8 to 21 days of age, were distributed in a completely randomized design with 6
treatments and 6 replicates of 15 birds each. A basal diet (based on corn-soy bean meal) was
formulated to provide of all nutrients according to Rostagno et al. (2011), with the exception of valine
level. The basal diet contained 2,991 kcal AMEn/kg, and L-glutamic acid was used to make it equal in
CP at 19% of the diet. To avoid excessive lysine, 90% digestible lysine (1.05%) was used. To obtain the
different valine to lysine ratio in treatment diets, crystalline L-glutamic acid in basal diets was gradually
replaced by crystalline L-valine, and six different ratios of digestible valine (% of lysine) of 70, 75, 80,
85, 90, and 95% were evaluated. To evaluate valine requirement of female broiler chicks based on
performance parameters, body weight and feed intake were recorded at the beginning and end of the
experiment, and the FCR was calculated. The result of this experiment showed that body weight gain
and FCR were significantly affected by the valine ratios in basal diet (P
EFFECT OF AMINO ACID LEVELS DURING BROILER BREEDER PULLET REARING ON REPRODUCTIVE PERFORMANCE AND FEATHERING UP TO 65 WK

E. Oviedo-rondon\*(1), A.Ortiz(1), Y.Matta(1), H.Cordova-noboa(1), G.Quintana-ospina(1), M.Alfaro-wisaquillo(1), J.Martinez(1), J.Yanquen(1), J.S. Hoyos-Torres(1), V. San Martin(1), J.Caldas(2)

(1) North Carolina State University, Raleigh, United States

(2) Cobb Vantress Inc., Siloam springs, United States

* Corresponding author: eooviedo@ncsu.edu

Dietary amino acid (AA) concentration during pullet rearing affects growth, development, and egg production. This experiment was conducted to determine the optimum AA levels for broiler breeder pullets from 5 to 24 wk of age to obtain the best egg production, hatchability, fertility, and feathering condition. Four AA levels were evaluated in the grower (2,700 kcal/kg ME) and developer (2,800 kcal/kg ME) phases. These diets contained balanced protein with digestible (dig) Lys of 0.48, 0.54, 0.60, and 0.66% for grower, and 0.51, 0.57, 0.63 and 0.69% for developer. Diets were formulated according to Cobb recommendations and ideal protein profile for each feeding phase. A total of 1,360 Cobb-500 slow-feathering pullets and 288 Cobb MV males were placed in an enclosed fan-ventilated 32-pen litter floor rearing house with 16 pens for females (85 pullets/pen) and 16 pens for males (18/pen). All pullets and males were fed one common starter diet in crumbles up to 4 wk. Grower diets were fed from 29 d up to 15 wk and developer mash feeds from 16 wk to 5% egg production. Feed amounts varied slightly (±3 g/d) among treatments in the developer phase to maintain BW close (±2%) to Cobb guideline (2018). Females and cockerels were fed in a 6/1 and 5/2 feed restriction program. At 21 wk, 7 males and 65 females were moved to 16 slat-litter floor laying pens, photostimulated with 14 h of light, and fed separately. After 5% egg production all hens were fed two common mash layer diets. Feed increments and reduction post-peak were made according to egg production by treatment. Egg production and mortality was recorded daily. From 27 to 65 wk, 180 eggs/pen were incubated every 2wk, and 30 eggs/pen used for egg quality evaluation. Data was analyzed in a completely randomized design with 4 AA levels and 4 replicate pens each. ANOVA and regression analyses were conducted. Dietary AA levels during rearing had a consistent quadratic effect (P < 0.05) on hen-wk (%) and total hen housed egg production indicating that the optimum level of dig Lys for grower and developer phases were 0.58% and 0.61%, respectively. The best average hen housed egg production observed upto 65 wk was 176.5 eggs and the worst 153.7 eggs. Eggshell strength and elasticity were affected (P < 0.05) by treatments up to 46 wk of age. Fertility was not affected (P > 0.05) by treatments, but hatchability increased linearly (P < 0.05) as AA during rearing increased up to 50 wk and decreased linearly during the last 15 wk of egg production. Feathering got worst after 52 wk and AA during rearing affected feathering and fleshing scores during lay in a quadratic (P < 0.05) manner. Feathering was related more to aging and fleshing than to AA levels during rearing. In conclusion, the optimum AA levels for the grower and developer phases are related to balanced protein with 0.58% and 0.61% dig Lys, respectively.
EFFECT OF ENERGY AND PROTEIN LEVEL UNDER STANDARD OR HEAT STRESS CONDITIONS ON GROWTH PERFORMANCE AND THE INCIDENCE OF MEAT MYOPATHIES

J. Diaz berrocoso*(1), A.Navarro villa(2), J.De los mozos garcia(3), A.Garcia ruiz(4)

(1)Trouw Nutrition, Madrid, Spain

(2) R&D Trouw Nutrition, El viso de san juan, Spain

(3)Trouw Nutrition Spain, Tres cantos, Spain

(4)Trouw Nutrition R&D, El viso de san juan, Spain

* Corresponding author: j.delosmozos@trouwnutrition.com

Energy and amino acids (AA) are the two nutrients with the largest contribution to the broiler feed price, animal performance and ultimately meat myopathies occurrence. However, the response of these nutrients on broiler performance is influenced by environmental conditions such as temperature and humidity. Therefore, a trial was conducted to determine the effect of dietary energy and digestible AA on growth performance, carcass yield and meat myopathies of broilers raised under recommended or heat stress (HS; high temperature and humidity) conditions. A total of 4160 broiler Cobb 500 were randomly assigned to 80 pens (52 broilers/pen, 50:50 males and females). Twelve treatments were arranged in a completely randomized block design using a 2 x 3 x 2 factorial arrangement with 2 dietary energy densities (AMEn; high vs. low), 3 energy to digestible lysine (dLys) ratios (low vs. medium vs. high) and 2 environmental conditions (Control vs. HS) and their interactions as main effects. Broilers were fed during 4 feeding phases (d0 to d 15; d15 to d29; d29 to d37; and d37 to 48d). Dataset of growth performance, carcass and breast meat yield and meat myopathies were analyzed using MIXED and GENDMOD procedure of SAS, respectively. Broilers raised under HS had 6% lower feed intake (FI) and 5% lower average daily gain (ADG; P<0.05) while feed conversion ratio (FCR) was not affected. Broilers fed with high AMEn/dLys (low Energy to dLys ratio) increased market weights (P<0.1), reduced FI (P<0.1), and improved FCR (P<0.05). Broilers fed high levels of energy had lower FI and better FCR (P<0.05). Carcass and breast meat yield increased with high dLys levels (P<0.05). Broilers growth under Control or HS responded similarly to energy and protein changes. Broilers fed low energy diets tended to reduce incidence of severe scores of white stripping whereas increased the incidence of poor cohesion meat. Wooden breast incidence was the highest with low AMEn/dLys (high dLys). It was concluded that the growth performance of the broilers raised under HS conditions performed worse than the birds raised in standard conditions independently of the energy and dLys level tested. In addition, the best growth performance, as well as the highest carcass and breast meat yield were obtained at low AMEn/dLys, disregarding the environment. Meat myopathies were affected by the energy and amino acids level being their incidence somewhat predictable.
THE RESIDUAL ANTINUTRITIVE POTENTIAL OF SOYBEAN PRODUCTS AFFECTS THE ESTIMATION OF STANDARDIZED ILEAL AMINO ACID DIGESTIBILITY BY MODULATING ENDOGENOUS PROTEIN SECRETION

S. Kuenz*(1), S.Thurner(2), D.Hoffmann(1), K.Kraft(3), M.Wiltafsky(4), K.Damme(3), W.Windisch(1), D.Brugger(5)

(1) Technical University of Munich, TUM School of Life Sciences, Chair of Animal Nutrition, Freising, Germany

(2) Bavarian State Research Center for Agriculture, Institute for Agricultural Engineering and Animal Husbandry, Freising, Germany

(3) Bavarian State Research Center for Agriculture, Department for Education and Poultry Research, Kitzingen, Germany

(4) Evonik Nutrition & Care GmbH, Hanau-wolfgang, Germany

(5) University of Zurich, Vetsuisse Faculty, Institute of Animal Nutrition, Zurich, Switzerland

* Corresponding author: sylvia.kuenz@wzw.tum.de

Soybeans are rich in trypsin inhibitors (TI), which impair protein utilization and thus animal performance. Hence, TI have to be reduced by heat and pressure. But a recent study has shown, that different processing techniques result in vast differences in qualities (Hoffmann et al., 2019). In this study, we examined how these quality differences in terms of trypsin inhibitor activity (TIA) affect the estimation of endogenous amino acid (AA) losses by linear regression extrapolation and, hence, the estimation of standardized ileal AA digestibility from these products.

For this experiment 45 soybean cakes (SBC) with a vast range of TIA (0.3-23.6 mg/g) were created using four different processing techniques (thermal, hydrothermal, pressure and kilning. These soybean cakes were then merged into a basal diet at two inclusion levels (15%, 30%). In this way 91 experimental diets (one basal diet as control) were designed. All diets contained 0.5% of titanium dioxide as indigestible marker. 5,490 1-day-old male broiler chickens (Ross 308) were used in four consecutive experimental trials. From d1 to d14 the birds were fed a commercial starter diet. On d15 the birds were allocated to pens with 10 birds each and the 91 experimental diets were fed ad libitum. On d22 the broilers were euthanized and the digesta of the terminal ileum was collected for determination of amino acid digestibility. For each SBC a digestibility coefficient (DC) was calculated. DC were analyzed using linear regression models defining TIA as main factor.

DC of every single AA declined significantly (p < 0.001) in a linear way with increasing dietary TIA. The depressive effect of TIA differed between single AA. Arg was the least affected AA with DC ranging from 44.8% - 96.5%. Digestibility of sulfur-containing AA was impaired most. Each increase of dietary TIA by 1mg/g reduced DC of Met and Cys by 2.36% and 2.31%.

This experiment showed that TIA has a linear effect on protein utilization, which means the presence of anti-nutritional substances must be considered using linear regression approach when estimating standardized ileal AA digestibility.

References

Hoffmann, D., S. Thurner, D. Ankerst, K. Damme, W. Windisch, and D. Brugger. 2019. Chickens' growth performance and pancreas development exposed to soy cake varying in trypsin inhibitor activity, heat-degraded lysine concentration, and protein solubility in potassium hydroxide, Poultry science. 98:2489-2499. doi: 10.3382/ps/pey592.
ID: 1147

EFFECTS OF DIETARY ENERGY AND PROTEIN LEVELS ON BROILER PERFORMANCE, CARCASS QUALITY AND WATER INTAKE

K. Bébin\*(1), M.Panheleux(1), S.Lecuelle(2), W.Lambert(2)

(1)CCPA, Janzé, France

(2)Ajinomoto Animal Nutrition Europe, Paris, France

* Corresponding author: lambert_william@eli.ajinomoto.com

Dietary protein and energy are the most expensive nutrients in broiler diets and their optimal supply must be continuously updated for modern broiler genetics. In addition, lowering dietary crude protein (CP) is well recognized to improve nitrogen efficiency, animal health and welfare.

In total, 1248 Ross 308 male broilers were allocated to six dietary groups varying in metabolizable energy (ME) and CP content, with 5 to 6 replicates per group. Birds were fed a common starter diet from 0 to 10 days. The control group (T1) contained 3000 kcal/kg ME, 20% CP and 1.05% digestible Lysine (dLys) in grower feed (10-21d) and 3050 kcal/kg ME, 18.5% CP and 0.98% dLys in finisher feed (21-35d). For T2 and T3, dietary ME was increased (3100 & 3200 kcal/kg in grower; 3150 & 3250 kcal/kg in finisher) with the same levels of dietary CP and digestible amino acids (dAA). From T4 to T6, the level of CP was decreased by 1% point per group in grower and 0.8% point per group in finisher (19, 18 & 17% in grower; 17.7, 16.9 & 16.1% in finisher) with the same levels of ME and dLys than T1, and with dAA kept above recommended levels for optimal performance. Average daily feed intake (ADFI), average daily gain (ADG) and feed conversion ratio (FCR) were recorded. Broilers were slaughtered at 35 days to measure carcass yield (CY), breast meat yield (BMY) and abdominal fat yield (AFY) for 40 broilers per group. Water intake was recorded and water to feed ratio calculated.

Increasing ME supply reduced ADFI from 0-35d, especially for T3 (111; 112 & 109 g/d for T1, T2 & T3) and in turn improved FCR (1.470, 1.457 & 1.428) but linearly reduced BMY and linearly increased AFY. No effect of dietary ME on ADG, CY or water intake was observed. Broilers fed lower dietary CP showed similar ADG, ADFI and FCR whatever the dietary CP content. Severe dietary CP reduction (T6) resulted in lower CY (72.0 & 71.7 in T1 & T6) and BMY (24.0 & 22.5 in T1 & T6). Dietary CP reduction also linearly increased AFY and linearly reduced water intake and water to feed ratio, an indicator of litter quality. The present study confirms that broiler may benefit from higher supply of energy and lower supply of protein, and that it is dependent on production criteria to optimize: performance, yields or welfare indicators.
THE INCLUSION RATE OF FREE AMINO ACIDS IS NOT A LIMITATION TO DECREASE THE PROTEIN LEVEL IN BROILER DIETS

P. Belloir*(1), M.Lessire(1), W.Lambert(2), E.Corrent(2), J.Van milgen(3), S.Tesseraud(1)

(1)INRAe, Nouzilly, France

(2)Ajinomoto Animal Nutrition Europe, Paris, France

(3)INRAe, Saint Gilles, France

* Corresponding author: sophie.tesseraud@inra.fr

The reduction of dietary crude protein (CP) is a major issue for sustainable poultry production from economic, environmental, and societal reasons. This requires the use of free amino acids (AA) added to diets to ensure a sufficient supply of indispensable AA. However, broiler performance is not systematically maintained when the CP content in the diet is drastically reduced, even when the indispensable AA requirements are apparently met. The aim of this study was to investigate whether the form of the provision of AA (i.e. as protein or as free AA) would limit performances in growing-finishing broilers. Thirty percent of the dietary CP fraction was provided by a highly digestible protein (casein) and/or a mixture of free AA whose digestible amino acid profile was exactly the same as that for casein, using graded levels of free AA inclusion in replacement of casein. Chickens (ROSS PM3) received one of the seven experimental diets (n=30 per treatment) during the whole experimental period (from d14 to d32) and were slaughtered at d32. Diets were formulated to be sub-limiting in digestible lysine (dLys) at 0.9%. The other AA were provided at levels to meet or exceed the recommendations, so that no AA other than Lys would limit growth. The AA digestibility and apparent metabolisable energy content did not differ between treatments. Increasing the level of free AA from 0 to 30% of the total CP content (i.e. 0.15 to 5.4% inclusion of free AA in the diet) resulted in reduced growth only in the diet with 30% free AA. The FCR was also degraded when the AA incorporation rate was greater than 20% free AA (i.e. 3.8% free AA in the diet). Chickens were able to adapt their digestive physiology (morphology and transporter expression) to increasing provision of free AA, at least for levels lower than 3.8% in the diet. Beyond this level, the absorption capacity may be affected, limiting the use of nutrients by the animal, which could partly explain poorer performance. In practice, the use of AA in free form should not be a limiting factor for the reduction of the protein content since the quantities for formulating low CP diets do not exceed 2% free AA in broilers.
EFFECT OF NUTRTIONAL MANIPULATION ON THE JUVENILE LOCAL KEETS (NUMIDA MELEAGRIS) RAISED IN TOGO

L. Lare*(1), E.Talaki(2), Y.Lombo(3), C.Sodjedo(4), D.Nideou(5), E.Oke(6), W.Pitala(7), K.Tona(5)

(1)Centre d'Excellence Régional sur les Sciences Aviaires, Lomé, Togo

(2) ESA/ Université de Lomé, Lome, Togo

(3) Institut Togolais de Recherche Agronomique (ITRA), Lome, Togo

(4)Ecole Supérieur d'Agronomie / Université de Lomé, Lome, Togo

(5)CERSA/ Université de Lomé, Lome, Togo

(6) Federal Univergy of Agriculture, Abeokuta, Nigeria

(7) Ecole Supérieure d'Agronomique (ESA)/ Université de Lomé, Lome, Togo

Traditional guinea fowl farmers use grains, agricultural by-products and termites without limit in the diets of the birds. This practice is inadequate for sustainable guinea fowl production as it provides insufficient or excess energy and protein. Reproductive performance in poultry is strongly linked to juvenile development. The most crucial and exigent period for rearing guinea fowl with energy and protein intake is early stage of growth (Lombo, 2018). The purpose of this study was to assess the effects of variation in energy and protein intake on local keets. A total of 600 local keets were allocated to 5 treatments groups (control, low-protein, high-protein, low-energy and high-energy) with 4 replicates of 30 keets each. Feed was offered ad libitum and feed intake was recorded daily. The birds were weighed weekly and mortalities were recorded. At 8 week, blood sample were collected for blood count and biochemical analysis. The data were processed by Graph Pad Prism 5.00.288 software and the ANOVA test was used to analyzed the results. The means were compared by the Tukey HSD test and the probability of P<0.05 was considered as the significance threshold. The results showed that the birds fed high-energy diet consumed less feed at all ages than those in other treatment groups. At 5 weeks onwards, body weights of keets fed low-energy and high-protein diets increased (p<0.05) more rapidly than those in control, high-energy and low-protein diets. Keets fed high-energy diet had a lower average daily gain than those in control, low-energy, high-protein and low-protein diets. Feed conversion ratio was similar across treatment groups. Keets fed high-protein and low-energy diets had less mortality. Blood parameters were not affected (p<0.05) by feed treatments. However, there was a significant difference (p<0.05) in the hematocrit according to feed treatements. This experiment showed that low-energy and high-protein diets improved body weights, average daily gain andreduced mortality of the birds.

Key words: Protein, energy, juvenile, local keets, Togo.

<footnote>* Corresponding author: nicody92@gmail.com</footnote>
INTERACTIVE EFFECTS OF DIETARY METHIONINE AND SELENIUM SOURCES ON BROILER GROWTH PERFORMANCE, CARCASS CHARACTERISTICS AND MEAT QUALITY

K. Decena*(1), R.Sulabo(2), M.Kumari(3), K.Israel(2), A.Sulabo(2)

(1) Animal Nutrition Division, Institute of Animal Science, College of Agriculture and Food Science, University of the Philippines Los Baños, Los baños, laguna, Philippines

(2) Institute of Food Science and Technology, College of Agriculture and Food Science, University of the Philippines Los Baños, Los baños, laguna, Philippines

(3) Department of Animal and Food Sciences, Faculty of Agriculture, Rajarata University of Sri Lanka, Anuradhapura, Sri Lanka

* Corresponding author: ksdecena@up.edu.ph

Broiler meat quality has shown to be affected by dietary methionine (Met) and also selenium (Se). However, the combined effect of both dietary Met and Se sources on broiler meat quality has not been previously investigated. Hence, this research focused on the effect of DL-Methionine (DL-Met) and DL-methylthio-butanoic acid (DL-HMTBA) in combination with Na selenite and 2-hydroxy-4-methylselenobutanoic acid (HMSeBA) on broiler growth performance, carcass characteristics and fresh breast and thigh meat quality and Se content. A total of 320 day-old, straight-run Cobb 500 broiler chicks were randomly allotted to 4 treatments following a randomized complete block design arranged as a 2 × 2 factorial. Factors were Met source (DL-Met and DL-HMTBA) and Se source in the mineral premix (Na selenite and HMSeBA). There were 8 replicate cages per treatment with 10 broilers in each cage. Broilers and feed refusals were weighed to calculate for ADG, ADFI and F/G. Meat quality of fresh breast and thigh meat were evaluated for pH, objective color, water-holding capacity, lipid oxidation, and Se content. The data were analyzed using the MIXED Procedure of SAS with cage as the experimental unit. There was a significant Met × Se source interaction for water-holding capacity (WHC) wherein the WHC of thigh meat was greater (P=0.016) if HMSeBA is used in DL-Met diets but lesser in DL-HMTBA diets. Broilers fed diets with DL-Met had greater feed/gain ratio (F/G) than those fed DL-HMTBA diets (P=0.009). Deboned thigh yield was greater by 5% (P=0.031) in broilers fed DL-Met diets than with DL-HMTBA. Consistently, the DL-Met fed birds had greater breast meat L* (P=0.011) and thigh meat pH 0h (P=0.027) and thigh meat pH 5h post mortem (P=0.002) than the DL-HMTBA. Na selenite elicited a 14% increase (P=0.037) in abdominal fat than the HMSeBA. Breast meat color, b* was greater (P=0.025) in HMSeBA fed broilers than the Na selenite. Lastly, selenium content in breast and thigh meat was greater (P<0.001) by 62% and 37%, respectively in HMSeBA fed broilers than Na Selenite. In conclusion, the dietary Met and Se source had independent effects on growth performance, carcass characteristics and meat quality of broilers. The WHC of thigh meat was greater if HMSeBA was used in diets containing DL-Methionine. Na selenite can be used as the Se source in DL-HMTBA diets without affecting the WHC of meat. HMSeBA improved the Se content of both the breast and thigh meat from broilers.
EFFECT OF SUPPLEMENTAL AMINO ACIDS TO LOW PROTEIN DIETS ON PRODUCTION PERFORMANCE, EGG QUALITY AND SERUM BIOCHEMICAL INDICES OF VANARAJA LAYING HENS

A. Panda\*(1), B.Sahoo(2), A.Kumar(2)

(1)I, Bhubaneswar, India

(2)ICAR-CENTRAL INSTITUTE FOR WOMEN IN AGRICULTURE, PLOT 50-51, MAUZA-JOKALANDI, NEAR KALINGA STUDIO, Bhubaneswar, India

\* Corresponding author: akpanda59@gmail.com

Among the nutrients needed by the bird, protein is one of the most important nutrient, which has a major influence on growth, feed utilization and production performance. The present study was conducted to study the effect of supplemental amino acids to low protein diets on production performance, egg quality and serum biochemical parameters of Vanaraja laying hens (32-48 weeks). A total of 120, Vanaraja laying hens were randomly distributed into four treatment groups with three replicates of 10 birds in a complete randomized design. Four different diets were formulated containing different levels of protein (13.5, 15, 16.5 and 18% CP) with similar energy (2600 ME kcal/kg). Lysine (0.75%) and methionine (0.35%) levels were kept constant in all the diets. Each dietary group were offered a measured quantity of feed daily. The egg production was recorded daily on individual pen basis. The eggs laid by the bird on individual pen basis during the last five days of each four weeks period were collected to measure the egg weight. Twelve eggs were randomly chosen from each dietary treatment from the eggs laid on 44th week to measure egg quality. At 48 weeks of age, about 2 ml of blood was collected from the wing vein of 12 birds (four birds from each replicate) per treatment. Data were subjected to statistical analysis under completely randomized design employing one-way analysis of variance and means of different treatments were compared with Duncan's multiple range tests. The hen-housed egg production, egg weight, egg mass per day and feed efficiency were not influenced by variation in CP contents of the diet. There was no influence of dietary protein levels on egg quality parameters like albumin %, Yolk %, Haugh unit and eggshell thickness except eggshell %. Significantly higher egg shell percentage was noticed in 16.5% and 18% CP diet (P<0.05) compared to either 13.5 or 15% CP diet. Serum biochemical parameters like total protein, globulin, cholesterol and creatinine, alkaline phosphatise (ALT) concentrations were not influenced by dietary protein levels. It is concluded that diet containing 13.5 % CP, 0.75 Lysine and 0.35% methionine is adequate for optimum production performance and egg quality of Vanaraja laying hens during 35 to 46 week of age.
OPTIMAL METHIONINE PLUS CYSTEINE REQUIREMENT IN BROILER STARTER DIETS SUPPLEMENTED WITH L-METHIONINE

M. Joske(1), D.Khan(2), A.Dedeurwaerder(1), B.Saremi*(2)

(1)Poulpharm BVBA, Izegem, Belgium

(2)CJ Europe GmbH, Schwalbach, Germany

* Corresponding author: Joske.Millecam@poulpharm.be

Methionine (Met) is the first limiting amino acid in broilers and together with cysteine (Cys) are two crucial amino acids to be met correctly. Methionine is supplemented in feed in powder form i.e. DL-Met and L-Met, or in liquid form (DL-Hydroxy analogue). The sulphur amino acid (SAA) requirements of broilers are defined in DL-Met supplemented diets. The objective of this trial was to investigate the optimal SAA requirement of broilers in the starter phase (first 10 days of life) in diets supplemented with L-Met. A basal diet containing 1.28% standardized ileal digestible (SID) Lys and 0.60% SID L-Met plus Cys (50% Met and 50% Cys; deficient in SAA) was created. L-Met was added to the basal diet to reach 0.65, 0.70, 0.75, 0.85, 0.95 and 1.05% SID L-Met plus Cys (6 replicates per treatment and 15 animals per replicate). The body weight (BW), daily weight gain (DWG), daily feed intake (DFI) and feed conversion ratio (FCR) were measured. Additionally, feed samples of the test diets were analysed to determine major nutrients including amino acids which appeared to be at or close to the formulated values. After 10 days on the test diets, significant differences (P < 0.05) were observed in all measured parameters. The optimal L-Met plus Cys requirements were determined using linear broken line and exponential asymptotic models. Model fit was evaluated based on the R2 of individual observations and graphical interpretation. The linear broken line model showed the best fit for BW, DWG and DFI. The models based on FCR showed a poor fit. Based on the fitted models, the optimal SID L-Met plus Cys level was found to be 0.69-0.70%. This new value is 20% lower compared to the recommendations of NRC and Ross 308 (0.90% and 0.94% SID Met plus Cys, respectively), which are defined based on DL-Met. Performance of broilers based on Ross 308 at day 10 of age is 321g BW. Herein, chickens at the requirements for the maximum performance needed 20% lower SID Met plus Cys while performing similarly (326 g BW). In conclusion, the lower Met plus Cys requirements might be due to the higher bioavailability of L-Met compared to DL-Met. Moreover, nowadays broilers are receiving lower crude protein diets, no AGPs, and have no access to animal protein sources. Hence, these changes in broiler feeding might be associated with the lower Met plus Cys requirements in trials based on L-Met as compared to the conventional requirements which has been based on DL-Met.
OPTIMAL METHIONINE PLUS CYSTEINE REQUIREMENT IN BROILER GROWER DIETS SUPPLEMENTED WITH L-METHIONINE

M. Joske(1), D.Khan(2), A.Dedeurwaerder(1), B.Saremi*(2)

(1)Poulpharm BVBA, Izegem, Belgium

(2)CJ Europe GmbH, Schwalbach, Germany

* Corresponding author: Joske.Millecam@poulpharm.be

Methionine (Met) is the first limiting amino acid in broilers. The requirement of Met together with cysteine (Cys) must be met correctly. Most of sulphur amino acid (SAA) requirement studies are defined with DL-Met supplemented diets. The objective of this trial was to determine the optimal SAA requirement of broilers in the grower phase (10-23 days of age) in diets supplemented with L-Met. A basal diet containing 1.15% standardized ileal digestible (SID) Lys and 0.55% SID Met plus Cys (50% Met and 50% Cys; deficient in SAA) was created. L-Met was added to the basal diet to reach 0.60, 0.65, 0.70, 0.80, 0.90 and 1.0% SID Met plus Cys (6 replicates per treatment and 15 animals per replicate). All birds were fed the same commercial starter diet during the first ten days of life meeting Ross 308 recommendations. The body weight (BW), daily weight gain (DWG), daily feed intake (DFI) and feed conversion ratio (FCR) were measured. Additionally, feed samples of the test diets were analysed to determine major nutrients including amino acids which appeared to be at or close to the formulated values. The optimal Met plus Cys requirements were determined using linear broken line and exponential asymptotic models. Based on the R2 of individual observations, the linear broken line model showed the best fit for BW, DWG and FCR. The models based on DFI showed a poor fit. The optimal SID Met plus Cys level was found to be 0.66% based on BW and 0.62-0.63% based on FCR and DWG. These new requirement values are more than 20% lower than the recommendations of NRC and Ross 308 (0.90% and 0.87% SID Met plus Cys, respectively), which are defined based on DL-Met. The suggested body weight of broilers for Ross 308 at day 23 of age is 1142g. In the present study, the comparative performance of the broilers (1191g BW) at day 23 of age was achieved with 20% lower Met plus Cys as compared to the standard recommendation. Herein, L-Met supplemented diets fed to broilers in grower phase caused a lower SID Met plus Cys requirements compared with the NRC and Ross 308 recommendations which are defined mostly using DL-Met based literature. Nevertheless, authors cannot exclude other confounding factors impacting the SID Met plus Cys requirement in broilers.
FEED TECHNOLOGY
ID: 488

HIGH-TEMPERATURE-SHORT-TIME CONDITIONING OF COMPOUND CORN-SOYBEAN DIET ENHANCES BROILER NUTRIENT DIGESTIBILITY

MA. Ebbing(1), N. Yacoubi(2), V.D. Naranjo Haro(2), W. Sitzmann(3), M. Gierus(1)

(1)University of Natural Resources and Life Sciences, Vienna, Austria

(2)Evonik Nutrition & Care GmbH, Hanau, Germany

(3)Amandus Kahl GmbH & Co. KG, Reinbek, Germany

* Corresponding author: marco.ebbing@boku.ac.at

Corn-soybean diets are used worldwide in the poultry industry. Conditioning these diets in a high-
temperature-short-time (HTST) environment, achieved by an expander, may improve broiler nutrient
digestibility of compound feed. The objective of the present study was to evaluate the effects of 4 HTST
intensities (0, 7, 10 and 20 kWh/t) performed in a pilot plant, on the broiler digestibility and performance.
For that, the treatments were applied to a corn-soybean, as basis diets. The diets were formulated
following the Ross-308 strain requirements. A three-feed phase trial was conducted from 1 to 35 days of
age in a completely randomized design with nine pens of 12 unsexed chickens each. Pen was the sampling
unit and all data were submitted to normality test, analysis of variance and regression test. Differences
between least square means were tested by Tukey test (P<0.05). Zootechnical performance was measured
on the overall period. Excreta samples for the determination of apparent total tract digestibility (ATTD) of
organic matter, total fat and AMEn were collected on days 32 and 33 of age. Ileal digesta for determination
of apparent ileal digestibility (AID) of starch and amino acids was collected at day 36 from 4 bird of each
pen and pooled. Titanium dioxide was used as indigestible marker. Performance wasn't affected by
expander intensities. In the average trial, the BWG at 35 days was 2588.10±79.42 g, FI was 3997.71±113.25
g and FCR was 1.532±0.044 g:g. In the one hand, these broilers had an ATTD of organic matter, total fat
and AMEn (y) affected quadratically by the HTST intensities (x), following the equations: y=-
0.00018x2+0.0034x+0.74, y=-0.013x2+0.28x+92.11, and y=-0.0051x2+0.13x+14.20 with maximal points:
9.44 kWh/t, 10.76 kWh/t and 12.74 kWh/t, respectively. Similar results were for the AID of starch (y), where
y=-0.007x2+0.17x+96.40, with 12.14 kWh/t as maximal value. In the other hand, amino acids digestibility
was not influenced by processing, with Met: 89.74±0.02 %, Lys: 82.43±0.03 %, Thr: 73.85±0.04 % and Arg:
87.20±0.02 %. In conclusion, increased expander intensities did not affect broiler's growth performance.
However, diets expanded with 9.44 to 12.74 kWh/t had higher energetic nutrient digestibility. In this way,
the absence of effects in AA digestibility suggest that the diets can be calculated with lower AMEn or
ingredients with lower energy content when HTST is used.
ID: 51

# IN VITRO INHIBITORY EFFECT OF A NEW THREE-STRAINS SELECTED PROBIOTIC ON SEVERAL SEROVARS OF SALMONELLA

C. Bostvironnois*(1), J.Schleifer(2), L.Skjoet-rasmussen(3)

(1)Chr Hansen, Arpajon, France

(2)Chr Hansen, Milwaukee, United States

(3)CHR HANSEN A/S, Hoersholm, Denmark

* Corresponding author: frcbos@chr-hansen.com

Bacillus are known to be capable of producing antimicrobial substances including e.g. bacteriocins which may be able to inhibit Gram-negative bacteria such as Salmonella (Abriouel et al., 2011). Reduced Salmonella colonization in liver and cecum has also been observed in Bacillus subtilis fed birds challenged with Salmonella Heidelberg (Hayashi et al., 2018) and Salmonella Enteritidis (Knap et al., 2011). The objective of the study was to evaluate the in vitro inhibitory effect of a new three-strain selected bacillus-based probiotic for poultry (GalliPro® Fit) on several Salmonella serovars using two different methods. The Salmonella enterica serovars tested were Albany, Braenderup, Enteritidis, Gallinarum, Heidelberg, Infantis, Kentucky, Mbandaka, Minnesota, Muenchen, Schwarzengrund, Stanley, Tennessee and Typhimurium.

The first method was an agar plate-based method. An agar plate was poured with pathogens on it. Wells were created into the plate. The bacillus-based probiotic was added to selected wells. After 24 hours of incubation, a zone of inhibition of up to 2 mm could be measured digitally.

The second method was a feed matrix-based methodology. The feed was autoclaved, then the probiotic was added at 10^5 CFU/g of feed. The pathogens S. Heidelberg or S. Typhimurium were added at 10^3 CFU/g of feed. During the 24 hours' time of incubation, samples were taken at different time points (4, 8 and 24 hours). Samples were added to plates, incubated and a counting of pathogens was conducted.

In these in vitro experiments the new bacillus-based probiotic was able to inhibit the growth of all Salmonella serovars belonging to different serogroups. The new bacillus-based probiotic inhibited type B, C, C1, C2, D Salmonella serogroups. When testing the probiotic vs. Salmonella spp. in an agar-based method, a radius of the inhibition zones of 1 to 2 mm was observed for all of them. When testing the probiotic vs. Salmonella spp. in a feed matrix, it inhibited the growth of S. Heidelberg (1.3 log inhibition after 4 hours, 4.6 log inhibition after 8 hours) and S. Typhimurium (1.6 log inhibition after 4 hours, 2.2 log inhibition after 8 hours). Both in-vitro methods confirm the growth inhibition activity on numerous Salmonella serovars by this new three-strain probiotic.
EFFECT OF AZOLLA ON EGG PRODUCTION PERFORMANCE AND CHOLESTEROL CONTENT OF EGG YOLK OF LAYING HEN

M.A Islam\*(1)

(1) Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, Bangladesh

\* Corresponding author: aminul_dgvc@yahoo.com

Abstract

Azolla pinnata is a small aquatic fern which flows on the surface of the water, rich in protein, essential amino acids (lysine, leucine, arginine, and valine), vitamins (vita A, Vita B12 and Beta-carotene), chlorophylls and minerals; Ca, P, Fe, Mg, Cu, Zn, Na, etc. It may be used in poultry diet as an abundant unconventional plant protein source which reduces the production cost and cholesterol and increases yolk color and profitability. Therefore, the present study was planned for assessing the effect of Azolla on egg production, profitability, egg yolk color, and cholesterol content of egg for producing quality, safe and profitable chicken eggs.

A total of 70 ISA-brown ready to pullets was distributed into 3 dietary groups; D1 (control-no Azolla) with 46 pullets, and D2 (diet with 4% sun-dried Azolla meal) and D3 (diet with 8% sun-dried Azolla meal) with 12 pullets each to investigate egg production performance and egg yolk color. Six experimental (2 eggs/diet) and 2 commercial farming (D4) eggs were tested to measure egg yolk cholesterol. The pullets were reared in the individual cage management system and fed iso-nitrogenous (iso-lysine) and an iso-energetic diet containing 17% CP and 2750 KcalME/kg up to 44 weeks of age. The collected data were analyzed using the Statistix10 computer package program. Cholesterol content of egg yolk was analyzed using a t-test.

Feed intake, FCR (Feed/kg egg), production cost, net-profit, and yolk color significantly differed among dietary groups (p<0.05 or p<0.001) > level of significance (Diet D₃ showed the highest egg mass, and the lowest feed intake and FCR (Feed/kg egg), followed by D1 and D2, respectively. Statistically similar egg, hen-day, and hen housed egg production were obtained among dietary groups. The lowest amount of production cost and the highest amount of net-profit was observed in D3, followed by D₁ and D₂, respectively. The deepest yellow yolk color was measured in D3, followed by D2 and D1, respectively. Therefore, Azolla significantly increased egg yolk color compared to the control diet. However, 8% of Azolla was much better than 4% Azolla in terms of increased yolk color. The lowest amount of cholesterol was determined in D₃, followed by D₂, D₁, and D₄, respectively (p<0.001) > the level of significance. Hence, commercial farming eggs showed the highest amount of cholesterol.

Therefore, Azolla may be considered as a novel and beneficial feed item for producing quality, safe and profitable eggs.

KEYWORDS: Azolla; cholesterol; egg production; hen; yolk color
THE IMPACT OF CONDITIONING TIME, STEAM PRESSURE AND SPEED OF COOLING ON THE THERMOSTABILITY OF A NOVEL HEAT STABLE 6-PHYTASE IN A LAYER FEED AFTER PELLETING.

L. Nollet(1), E.Vanderbeke*(1), M.Eeckhout(2)

(1)Huvepharma NV, Antwerp, Belgium

(2) Ghent University, Faculty of Bioscience-engineering, Ghent, Belgium

* Corresponding author: lode.nollet@huvepharma.com

The aim of the study was to investigate the impact of steam pressure, conditioning time and speed of cooling on the thermostability of a novel E. coli phytase (OptiPhos® Plus CT, Huvepharma NV). A wheat/corn based layer feed was supplemented with phytase to yield a final concentration of ± 1000 FTU/kg. The feed was prepared in a vertical mixer (700 litres) and conducted to a conditioner unit. The conditioner was a fast rotating paddle mixer with a capacity of 20 litres in which steam is added. This steam is produced via an electric steam generator at 9 bar and is dewatered and reduced to a working pressure of the desired bar before entering the conditioner. The residence time is controlled manually. The pellet mill is a Simon Heessen Labor Monoroll type (Adritz Sprout, Netherlands) with a 4x50 mm die. The conditioning was performed at 85°C, but with different residence times and steam pressure: 30 sec. & 1.5 bar saturated steam (steam temperature: 127°C), 30 sec. & 3 bar saturated steam (steam temperature: 140 °C), 200 sec. & 1.5 bar saturated steam (steam temperature: 127°C). Cooling and drying was performed by an ambient flow through air cooler in 2 ways: instantly or slowly. Instant cooling means each subsample (10) is cooled instantly after leaving the die, while slow cooling means the subsamples (10) are first pooled during 2 to 3 minutes of production and then brought in the cooler. In both cases the residence time in the cooler was equal to 15 minutes. The recovery was determined as (phytase in supplemented pellet – phytase in blanc pellet)/(phytase in supplemented mash – phytase in blanc mash). Results showed that condition time had no impact on heat stability of the phytase, yielding 94.9 % and 92.9 % at 30 sec conditioning with respectively 1.5 and 3 bar steam pressure, and 93.2 % at 2 min. conditioning and 1.5 bar steam pressure. Speed of cooling had a large impact on the recovery as slow cooling brought recoveries down to 86.8 and 82.6 % at 30 sec conditioning and 1.5 respectively 3.0 bar steam pressure, while recovery was 76.5 % at 2 min. conditioning and 1.5 bar steam pressure. It can be concluded from this trial that conditioning time and steam pressure did not have much impact on the heat stability of the phytase, while cooling had a large impact. Therefore it is also advisable in pelleting studies to detail better, or to uniform, the cooling of (sub)samples.
EFFECTS OF PRONUTRIENTS (BOTANIC MOLECULES) ON PERFORMANCE AND COMPARISON WITH BUTYRATE IN BROILERS

J. Pie*(1), T.Chowdhury(2), S.Dey(3), A.Dhara(3), D.Diez(1), C.Domenech(1), S.Haldar(3), A.Sarkar(3)

(1)Biovet S.A., Constanti, Spain

(2) Doctor's Agro-vet Ltd., Dhaka, Bangladesh

(3) Agrivet Consultancy Pvt Ltd., Lake town, kolkata, India

* Corresponding author: jpie_biovet@yahoo.com

Pronutrients are active molecules from plant extracts that naturally improve the physiology of their target organs.

An experiment was conducted to determine the effects on performance and carcass yield of pronutrients, butyrate and the combination of both in broilers. Pronutrients and butyrate are described as natural growth promoters and can replace AGPs.

320 male Vencobb broilers were distributed into four treatment groups with 80 birds each: control (T1); a group receiving pronutrients in feed at 0.5 kg/t (T2); a group receiving butyrate in feed at 0.3 kg/t (T3); and a group receiving the combination of pronutrients + butyrate at the mentioned doses (T4). Birds received the treatment continuously from day 1 until day 35 of age (end of trial). All diets were fed ad libitum. Data were statistically analyzed by multi-variate ANOVA in GLM of SPSS (v 26.0). Results showed that pronutrients (T2) improved body weight (BW), whether alone (+3%) or in combination with butyrate (+3.9%), compared to the control (T1). Butyrate alone (T3) showed no positive effect on BW. Feed conversion (FCR) was improved in all treatments (T2, T3 and T4), compared with the control, and pronutrients (T2) improved it significantly (-4.9%, P

In conclusion, pronutrients in feed can be used to improve productive performance in broiler chickens, as it showed a trend to increase BW, decrease mortality, and significantly improved FCR. It was more effective than butyrate. Additionally, no interaction was observed between pronutrients and butyrate, so they can be added together to broiler diets.
ID: 283

GROWTH PERFORMANCE, INTESTINAL MORPHOMETRY, AND BLOOD SERUM PARAMETERS OF BROILER CHICKENS FED DIETS CONTAINING INCREASING LEVELS OF WHEAT BRAN WITH OR WITHOUT EXOGENOUS MULTI-ENZYME SUPPLEMENTATION

F. Idan\*(1), T. Nortey(2), C. Paulk(1), C. Stark(1)

(1) Kansas State University, Manhattan, kansas, United States

(2) University of Ghana, Accra, Ghana

* Corresponding author: kingkizo@ksu.edu

Wheat bran (WB) is an alternative ingredient that could be utilized as an energy and protein source partial replacement of corn and soybean meal in poultry feed formulation. The objective of the study was to investigate the effects of multi-enzyme supplementation (XGP) on the growth performance, intestinal morphometry, and blood-serum parameters of broilers fed diets containing increasing levels of WB. A total of 720 Cobb 500 seven-day-old chicks were assigned to 1 of 8 treatments to obtain 15 chicks per replicate and 6 replicates per treatment. A 2 x 4 factorial arrangement with 2 levels of multi-enzymes (0 or 360 g/ton) and 4 levels of WB (0, 4, 8, or 12%) were used. The multi-enzyme premix consisted of Xylanase, β-Glucanase, and Phytase (XGP). Bodyweight and feed intake were determined at 14, 28, and 42 d and used to calculate ADG, ADFI, and FCR per pen. There was a WB x XGP interaction (P<0.01) for the overall BW and ADG of broilers. Broiler ADFI was not affected by dietary treatments. Increasing the WB level linearly increased (P=0.003) the overall FCR. Besides, WB x XGP interaction (P<0.01) was observed for villi height (VH) and villi height to crypt depth ratio (VH: CD). XGP increased (P<0.01) high-density lipoprotein cholesterol (HDCL) but decreased cholesterol (CHOL). Feeding broilers diets containing increasing levels of WB with or without XGP resulted in similar BW, ADFI, and FCR as broiler control diets. Increasing the WB level linearly decreased overall BW and made FCR of broilers poorer. Broilers fed XGP had increased VH, VH: CD, and HDCL but decreased and CHOL.
WATER ACIDIFIER ENHANCES FEED EFFICIENCY AND BREAST YIELD OF BROILERS

G. Heim*(1), S.Rama Rao(2), L.Pineda*(3), S.Kadari(4), B.Brutsaert(1), Y.Han(1)

(1)Trouw Nutrition, Amersfoort, Netherlands

(2) Sri Ramadhootha Poultry Research Farm Pvt Ltd, Hyderabad, India

(3)Trouw Nutrition, Bangkok, Thailand

(4)Trouw Nutrition, Bengaluru, India

* Corresponding author: giseli.heim@trouwnutrition.com

Water acidifier enhances feed efficiency and breast yield of broilers

Giseli Heim1 S.V. Rama Rao2, L.M. Pineda3, S. Kadari4, B. Brutsaert1, Y. Han3

1Trouw Nutrition Innovation, The Netherlands

2Sri Ramadhootha Poultry Research Farm Pvt Ltd

3Trouw Nutrition R&D, The Netherlands

4Trouw Nutrition, India

ABTSRACT

The current trend in the global poultry industry is towards a reduction in antibiotic use during production. To maintain the same level of zootechnical performance achieved with antibiotic growth promoters (AGP), several feed additives are being marketed as suitable AGP alternatives. This study was designed to investigate the efficacy of water acidifer (SPH) based on a synergistic blend of free and buffered short-chain fatty acids on growth performance and carcass yield of broiler chickens as AGP replacement. One-day-old male broiler chicks (n=1300) were randomly assigned to one of four treatments with 13 replicate pens of 25 birds each. The treatments included a negative control (NC, a basal diet without AGP), a positive control (AGP, as NC + Bacitracin methylene disalicylate (BMD; 500 g/T in phase 1-3) and Virginiamycin (45 g/T in phase 3); water acidifier (SPH, as NC + 2L SPH/1000L water), and water acidifier + AGP (SPH+AGP, as PC+SPH). All chicks were reared in floor pens with a deep litter system in an open-sided poultry house and were provided with a three-phase corn-soy based diet. The body weight (ADG) and daily feed intake (ADFI) were recorded at d1, 14, 28, and 42 and the feed conversion ratio (FCR) was calculated. The carcass yield (percentage of breast weight (without skin) over the hot carcass weight) and dressing percentage were evaluated and the feed cost per kg live weight was calculated (based on the cost of the product and market conditions at the time of the study) on d42. Data were analyzed using the GLM procedure in SAS and Tukey's range test was used to determine significance between treatment means (P

In all parameters examined, no significant difference was observed between the AGP, SPH and the combination of SPH+AGP treatments (P>0.1).

In contrast to the NC, the FCR reduced significantly by -7.2 points with the supplementation of SPH+AGP (1.697 vs 1.615, P=0.02), while the effect of SPH (1.650) or AGP (1.664) was similar and intermediate between SPH+AGP and the NC (P>0.05). The breast yield increased significantly by +1.9% compared to the NC (26.32 vs 24.39%, P=0.04). The dressing percentage, livability and cost to produce kg LW were not affected by the treatments (P>0.05).

In conclusion, the study demonstrated that water supplementation with SPH or in combination with AGP (SPH+AGP) is as effective as AGP in enhancing breast meat yield and improving feed efficiency of broiler chickens.

Keywords: additives, antibiotic-free environment, antibiotic resistant
ID: 431

EFFECTS OF NATURALLY CONTAMINATED DIETS WITH FUSARIUM TOXINS AND ADDED ANTI-MYCOTOXIN AGENT ON PERFORMANCE PARAMETERS AND LIVER HEALTH OF LAYING HENS

M. Janječić*(1), D.Bedeković(2), K.Salajpal(2), I.Kos(2), G.Kiš(2), M.Caballero(3), Z.Janječić*(1)

(1)University of Zagreb Faculty of Agriculture, Zagreb, Cuba

(2)University of Zagreb Faculty of Agriculture, Zagreb, Croatia

(3)EW Nutrition, Bremen und umgebung, Germany

* Corresponding author: mihael.janjecic@gmail.com

T-2 is a mycotoxin classified as trichothecene, produced by Fusarium molds infecting cereal crops, and found frequently in raw materials used for animal feeds. The toxic effects of DON and T-2 on poultry are shown mainly in the liver, the gastrointestinal tract and the immune system. Liver function is crucial for egg production and when liver function is impaired egg quality can be detrimental and when the exposure is chronic egg production can also be affected. 90 Lohmann Brown laying hens were assigned randomly to three groups (10 repetitions - 3 hens each) to evaluate the effects of 12 weeks' exposure to multiple Fusarium toxins through feed, and its amelioration by an anti-mycotoxin agent. The groups included: Control (C) fed a standard diet, very low mycotoxin load: DON ~100ppb; Fumonisin B1+B2 ~400ppb; Zearalenone ~20ppb; Low (L) fed a contaminated diet with T-2 ~90ppb, DON ~100 ppb, Fumonisin B1 ~1000ppb, Fumonisin B2 ~250ppb and Zearalenone ~50ppb & another one fed with the contaminated diet adding 0.15% of an anti-mycotoxin agent (LM). Performance was measured daily, feed intake and egg quality weekly. Egg quality included weight, Haugh units, eggshell strength, eggshell thickness, shape index, and relative albumen, yolk and eggshell weights. Blood samples were taken for Ca, P, liver health and stress indicators. Liver samples (five birds per group) were semi-quantitatively scored on a scale of 0-5 for color, presence of haemorrhages (macroscopic); tissue was taken for histopathology. The data was fitted by nonlinear growth models within R environment, by R "nlseasy" package. There were no significant differences between the fitted growth curves for the treatments, although in the last 6 weeks, L showed a tendency for lower feed intake and laying smaller eggs. No differences in liver health indicators (AlbG, AlkP, ALT and AST, GGT and GLDH), stress indicators (H/L ratio), blood Ca and P levels, and liver macroscopic were found. Histopathologic examination showed mostly scant to mild multifocal interstitial and perivascular infiltrates of lymphocytes in all liver tissue samples. It can be concluded that the mycotoxin challenge was not able to reduce welfare, health performance and egg quality of laying hens during the trial period but longer exposure to mycotoxins challenge without binders can result in reduced feed intake and smaller eggs.
THE EFFECT OF PRE-PELLETING WHOLE CORN INCLUSION ON GROWTH PERFORMANCE, PROCESSING YIELD, AND ORGAN DEVELOPMENT OF BROILERS

F. Ovi*(1), W.Pacheco(1), C.Starkey(1)

(1)Auburn University, Auburn, United States

* Corresponding author: fko0001@auburn.edu

Previous research suggested that whole grain inclusion can reduce grinding costs and improve broiler performance. Two studies were conducted to evaluate the effect of whole corn inclusion pre-pelleting on growth performance, processing yield, and organ development of broilers. Both experiments consisted of 4 dietary treatments with 10 replicate pens per treatment and 25 broilers per pen. In first experiment treatment diets consisted 0, 2.5, 5, and 7.5% whole corn that replaced ground corn and was provide from 1 to 42 d of age. In second experiment, 0, 3, 6, and 9% whole corn was included from 14 to 42 d of age, following a common starter diet from 1 to 13 d. The starter diets were fed in crumbled form and the grower and finisher diets in pelleted form. In both studies, feed intake and BW were determined at 14, 28, and 42 and FCR adjusted by adding the weight of the mortality to the BW of live birds. At 43 d, 10 birds/pen were processed for yield determination. In the second study two-broilers per pen were euthanized by CO2 asphyxiation and used to determine the weight of crop, proventriculus, gizzard, liver and ceca and expressed as a ratio of total live body weight. Data were statistically evaluated using ANOVA procedure and means were separated by Tukey HSD procedure. In first experiment whole corn inclusion significantly improved FCR from 28 to 42 d of age (1.94 vs. 2.00, P<0.05). Birds fed diets with 5% of whole corn had higher breast meat weight (292 vs. 284 g, P0.05) from 14 to 42 d of age. However, birds fed diets with 9% whole corn had higher carcass yield (P0.05). But the relative proventriculus weight decreased with increasing level of whole corn inclusion(P=0.036, 0.302 vs 0.346). Results of these studies indicated that 5% whole corn can be included in starter feed and 9% in grower and finisher feeds without a negative effect on broiler performance.
SUPPLEMENTATION OF PROTEASE IN A LOW PROTEIN DIET ON GROWTH PERFORMANCE IN BROILERS
FOR 35 DAYS

C. Hyunmin*(1), J.Hong(2), Y.Kim(2), S.Nawarathne mudiyanselage(2), C.Lee(3), J.Heo(2)

(1)Chungnam National University, Daejeon, South Korea

(2)Chungnam National University, Republic of Korea, Daejeon, South Korea

(3)Kemin Animal Nutrition and Health, Asia Pacific, Seoul, South Korea

* Corresponding author: wh1tp@naver.com

Research on exogenous enzymes is being actively conducted on broilers to improve the protein utilization and subsequent reduction of nutrients in the animal waste generated from the feed. Endogenous proteases synthesized and secreted in the gastrointestinal tract (GIT) are known to be sufficient for the protein utilization in the diet (Nir et al., 1993). However, reports have demonstrated that considerable amounts of protein pass through the GIT rather than completely digested in broilers. Undigested proteins in the GIT can improve the digestibility by endogenous proteases. Therefore, the objective of this study was to evaluate the effects of the endogenous protease on growth performance with low protein and amino acid diet.

A total of 448 one-day-old Ross male broiler chickens were allocated according to completely randomized design into 64 pens. Eight dietary treatments were practiced and each treatment consisted eight replications with seven birds in each cage. Dietary treatments were: positive control with standard diet (PC), PC + protease (Kemin; 150g/ton; PC-150), negative control with CP 0.5% and AA 2% down from PC (NC1), NC1 + protease (150g/ton; NC1-150), NC1 + protease (300g/ton; NC1-300), negative control with CP 1.0% and AA 4% down from PC (NC2), NC + protease (150g/ton; NC2-150), NC2 + protease (300g/ton; NC2-300). Corn and soybean meal control diets were formulated to meet the Ross 308 nutrition specification. Diets were provided on an ad-libitum base in a mash form. The protease was top-dressed onto the diets in order to make eight treatments. Average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) were measured for 5 weeks. Data were subjected to analyse using one-way ANOVA in SPSS.

Results revealed that the supplementation of protease into low CP, AA density diets improved (P<0.05) the daily gain (PC-150: 2.2% than PC; NC1-150: 1.7%, NC1-300: 3.8% than NC1; NC2-150: 1.4%, NC2-300: 7.2% than NC2) and feed efficiency (PC-150: 7.2% than PC; NC1-150: 2.2%, NC1-300: 3.9% than NC1; NC2-300: 3.8% than NC2) of the chickens compared to the chicken fed with NC1, NC2 diets from hatch to 35 days.

In conclusion, our results indicated that the supplementation of protease in to the reduced CP, AA diets has the ability to improve the growth performance of broiler chickens by compromising the impact of low CP, AA in the diets.

Nir I Z Nitsan and M Mahagna. 1993. Br. Poult. Sci. 34:523-532.
NATURAL FEED ADDITIVE FORM AGRICULTURAL BIOMASS WASTE: MICROENCAPSULTATED PYROLIGNEOUS ACID POWDER FOR BROILER REARED IN THE TROPICAL CLIMATE AREA

Y. Theapparat\*(1), N.Roekngam(1), S.Khongthong(2), K.Soisuwan(3), D.Faroongsarng(1)

(1) Drug Delivery System Excellent Center, Faculty of Pharmaceutical Science, Prince of Songkla University, Songkhla, Thailand

(2) Faculty of Veterinary Science, Rajamagala University of Technology Srivijaya, Nakhon sri thammarat, Thailand

(3) Department of Animal Science, Faculty of Agriculture, Rajamagala University of Technology Srivijaya, Nakhon sri thammarat, Thailand

* Corresponding author: yongyuth.theap@gmail.com

One approach to utilize agricultural biomass waste is carbonization, which generates organic charcoal as the main product and pyroligneous acid (PA) as a by-product. The microencapsulated powder of PA prepared from the carbonization of agricultural biomass waste so-called natural feed additive (NFA) was developed. It composes with rich phenolic compounds and shot and medium chain fatty acids as the main components with a concentration of 72.94-94.91 mg/g and 63.18 mg/g, respectively. The stability testing of the encapsulated powder shown more stable for animal feed industrial applications. The effect of NFA supplement on growth performance and healthy improvement in broilers was investigated. 400 one-day-old male broiler chicks (Ross 308) were allocated into 4 treatment groups including (1) negative control (no supplemented NFA) (2) supplemented NFA 1,500 mg/kg (3) supplemented NFA 3,000 mg/kg (4) supplemented NFA 4,500 mg/kg. Each treatment was divided into 5 replicates of 20 chicks each (20 males). The results found that it was not improving the growth performance in broilers. However, stress and inflammatory levels in broilers feed with NFA treatments were suppressed by which the blood cortisol hormone and IL-1b, IL-8, IL-10, TLR-4 and IFN-g in an intestinal tract were significantly lower than that the control treatment (P<0.05). Besides, it also increasing volatile fatty acid content in Ileum and caeca in broiler with statistical significance when compared with control treatment. Moreover, it stimulated immunological response by which the IgG, IgM, and IgA in plasma of broilers feed with NFA have a significantly higher concentration than that the control treatment (P<0.05). The intestinal histology showed that male chickens fed diets supplementing 1500-4500 mg/kg NFA had significantly higher villus height and villus height:crypt ratio of duodenum and jejunum as compared to the control group (P<0.05). The gut microbiota analysis showed that supplement the NFA can improving the actinobacteria in the intestinal tract for boiler. It can be implied that the NFA prepared for this study has the prebiotic characteristic. Overall, it could be concluded that the feed additive product prepared from agricultural biomass waste in microencapsulated powder has the potential for application as a natural feed additive for boiler with integrated functions.
EFFECT OF YEAST FRACTION SUPPLEMENTATION ON INTESTINAL ENVIRONMENT IN BROILER CHICKS
WITH DEXTRAN SULFATE SODIUM

T. Bungo\*(1), T.Hirota(2), Y.Ouchi(2), T.Nii(2), L.Faivre(3), C.Janvier(3), T.Ide(4)

(1)Hiroshima University, Higashi0hiroshima, Japan

(2)Hiroshima University, Higashi-hiroshima, Japan

(3)Phileo Lesaffre Animal Care, Singapore, Singapore

(4)YPTECH Co., Ltd., Tokyo, Japan

* Corresponding author: bungo@hiroshima-u.ac.jp

It is known that yeast fraction (YF) works as prebiotics and improves gut health by adsorption of harmful bacteria and maintenance of barrier function in animals. The objective of the study was to investigate the protective effects of YF added to diet on the intestinal environment in broiler chicks challenged with dextran sulfate sodium (DSS), an agent to cause gut inflammation. Chicks were separated in two groups: one control group fed with a basal diet and another group fed with the basal diet supplemented with 0.025% YF. At 14 days of age, half of the birds in the birds in the control group (CC) and half of the birds in the yeast fraction group (YC) were given saline (0.1 mL). The other half of control group (CD) and yeast fraction group (YD) were orally gavaged with DSS (1 mg/0.1 mL). The number of chickens in each group was n = 6. After 6 h, all chicks were hyperanesthetized, and their jejunum tissues and cecal contents were collected. Samples were stored at -80°C prior to the RNA or DNA isolation. As for the gene expressions in jejunum tissue, significant increases were found on mucin-1, claudin-5, and IL-8 in YF group compare to other groups. Same effects were measured in YD group with the mRNA levels of mucin-1, and IL-8. Regarding the cecal microbiota, ratio of Bacteroidesin CD group tended to be higher than other groups. These findings suggest that YF may work against the substance, which has the ability to disturb the intestinal environment, and helps to recovery any damage induced by a harmful material in broiler chicks immediately.
EFFECT OF COMBINATION OF HYDRATED SODIUM CALCIUM ALUMINOSILICATE (HSCAS) AND CLINOPTILOLITE AS ENDOTOXIN BINDER ON BROILER PERFORMANCE AND INTESTINAL HISTO-MORPHOLOGY

S. Sooryan\*(1), S.Murugan(2), B.Chacko(2), C.V. Rajani(2), H.Patki(2), S.Joseph(2)

(1)Kerala Veterinary and Animal Sciences University, Pookode, Wayanad., Chikamagalur, India

(2)Kerala Veterinary and Animal Sciences University, Pookode, Wayanad, Wayanad, India

\* Corresponding author: sathyasooryan25@gmail.com

Poultry is being exposed to endotoxins released by the death of Gram-negative bacteria present in the normal intestinal flora and the environment. Sub-therapeutic antibiotic usage as a growth promoter is one of the causes of the increased release of endotoxin from these bacteria (Shinozuka et al. 2007). In the current research, a combination of hydrated sodium calcium aluminosilicate (HSCAS) and clinoptilolite was evaluated as an endotoxin binder (EB) in broilers. The EB was used alone and along with an antibiotic (AB) to assess its effect on broiler growth performance, intestinal histo-morphology and techno economics of EB inclusion.

Two hundred day-old Vencobb-400 broiler chicks were divided into five groups with four replicates of ten chicks in each. The experimental treatment consisted of G1-fed with basal diet (BD) + 0.5 kg/ton AB, G2-BD + 2.0 kg/ton EB, G3-BD + 2.5 kg/ton EB, G4-BD + 1.0 kg/ton EB + 0.5 kg/ton AB and G5-BD + 1.5 kg/ton EB + 0.5 kg/ton AB. The daily feed intake and weekly body weight were recorded and six birds from each group were sacrificed at every phase (pre-starter, starter and finisher) for intestinal studies. The data obtained was subjected for Duncan one-way ANOVA using SPSS-IBM 24.0 version software. Fourier Transform Infrared spectrum of EB was analysed.

The resulted body weight gain (BWG) and feed efficiency ratio (at 42nd day) compared between EB alone and EB with AB groups demonstrated that either EB at 2 kg/ton or EB at 1.0 kg/ton along with AB were significantly (Pnd day was significantly (P
USE OF A PHYTOGENIC BLEND OF CINNALMALDEHYDE AND THYMOL ALONE OR IN COMBINATION WITH A C=BACILLUS PROBIOTIC IMPROVES PERFORMANCE OF BROILERS UNDER HIGH CHALLENGE SITUATION

C. Evans(1), A.Ghane*(2), T.Stormink(3), F.Sidiq(4), K.Gibbs(1)

(1)DuPont Animal Nutrition, Marlborough, United Kingdom

(2)DuPont Animal Nutrition, Bangkok, Thailand

(3)DuPont Animal Nutrition, Leiden, Netherlands

(4)DuPont Animal Nutrition, Jakarta, Indonesia

* Corresponding author: ceinwen.evans@iff.com

The current study looked at the effects of including a phytogenic blend (EO), alone or in combination with a Bacillus based probiotic (EOPRO) or an AGP in broilers fed a diet containing a phytase/carbohydrase combination with associated full matrix applied under challenged conditions. Total of 2160 day-old Lohman Indian River chicks was randomly allocated to 5 dietary treatments with 12 replicates (36 birds/pen). Birds were fed test diets in 2 phases: starter (D1-21) and grower/finisher (D22-35). Control diets were formulated to meet the birds nutrient requirement allowing for nutrient matrix of enzymes. All diets contained Buttiauxella phytase (Axtra® PHY) at 1000 FTU/kg of feed and a combination of xylanase, amylase and protease (Avizyme® 1505X) at 200 g/tof feed. One group of birds were fed the control diet and given no challenge. Challenged birds (CC) were inoculated with C. perfringens and E. coli. The CC were fed either unsupplemented or supplemented with Antibiotic Growth Promotor (BMD) at 50g/t, Phytogenic blend (EO) at 100 g/t, or a combination (EOPRO) of EO at 100 g/t and a 3 strain Bacillus probiotic at 60 g/t of feed. Gut samples were taken on days 21 and 35, from 24 birds/treatment. Lesions were scored in duodenum (E. acervulina), ileum (E. maxima) and caeca (E. tenella). Results confirmed the applied enzyme matrix as the unchallenged birds fed the enzyme containing control diet hit their production targets. Supplementing the diets of CC with EO and EOPRO treatments resulted in numerically improved performance. Average daily gain was numerically improved with EO and EOPRO compared to the CC by 1.31% and 2.45%, respectively. FCR was numerically improved by 3 points with both EO and EOPRO treatments vs. CC and was not significantly different to the UC birds. In contrast the AGP treatment did not result in any performance improvements in the CC, with both the EO and EOPRO treatments numerically improving performance versus the AGP. Broiler Index was numerically improved by 4.36% and 5.23% by EO and EOPRO treatments compared to the CC. Gut health scores did not differ between treatments. Results showed that full matrix implementation for combination of Phytase (Axtra PHY) and mixed enzymes (Avizyme) at 126 kcal/Kg ME was the proper level based on the diets' specifics. The EO and EOPRO treatments proved to be effective options to replace AGP and maintain or improve performance under challenged conditions.
EFFECT OF GRAIN SORGHUM PARTICLE SIZE, NUTRIENT LEVEL, AND PHYTASE DOSING ON THE PERFORMANCE OF BROILER CHICKS TO 18 DAYS OF AGE

S. Beyer*(1), C.Delfelder(1)

(1) Kansas State University, Manhattan, United States

* Corresponding author: sbeyer@ksu.edu

Broiler diets require cereal grains to be finely ground to improve pellet quality, which optimizes feed intake and growth rates. This grinding decreases the work necessary by the gizzard resulting in reduced physical exertion and atrophy of gizzard mass. Grain sorghum is more apt to produce crops in dryland farming but must be properly processed to optimize nutrient retention when fed to broilers. Because the gizzard functions to grind and mix digesta for processing in the small intestine, a study was conducted to test the effect of combinations of grain sorghum particle size (PS), nutrient density and phytase enzyme levels on broiler chick performance. Whole grain sorghum was ground using a hammer mill to PS of 378 nm (small) and 747 nm (large). Control rations were formulated to meet the recommended nutrient requirements of Ross male chicks (Positive control, PC) with rations made using either small or large sorghum particles then fed in mash form. A set of negative control rations (NC) were formulated to 3% less energy and protein than the PC and with the available P (aP) reduced to 0.4%. Each NC diet was manufactured with small or large PS sorghum and supplemented with 0, 500 or 1500 FYT of phytase (DSM Ronozyme HiPhos 2700). The battery trials were conducted with 6 chicks per pen and 6 replications of each treatment. Performance data and gizzard weights were recorded at 18d. Data were analyzed as a factorial arrangement using main effects of diet (PC vs NC), PS (small vs large) and phytase level (0, 500 or 1500 FYT) using SAS (9.4) with significance levels set at P≤0.05. The results showed a significant main effect of PS to increase total body weight gain (BWG), gizzard weight (GW) and improve feed conversion (FC) when fed diets manufactured with large sorghum PS. These parameters were significantly reduced by diets limited in nutrients and aP but improved with increasing phytase. When the NC rations were supplemented with phytase, BWG, GW, and FC were significantly improved (P≤0.05) with increasing enzyme levels with the optimum performance observed for the group fed the large PS at 1500 FYT of phytase. The results indicate that maintenance of gizzard mass is important to optimize the BWG and FC of broiler chicks when feed rations are supplemented with increasing levels of phytase enzyme. Although pellet quality improves by reduction of PS, research is needed to determine optimal PS for the maintenance of desired gizzard function in broilers.
# EVALUATION OF DIETARY SUPPLEMENTATION OF A NOVEL MURAMIDASE TO WHEAT-SOYA DIETS ON BROILER PRODUCTION EFFICIENCY IN THE PRESENCE OF FEED ENZYMES AND COCCIDIOSTAT

J. Houdijk*(1), I.Eising(2), O.Olukosi(1), R.Lopez-ulibarri(3), F.Khattak(1), E.Perez-calvo(4)

(1)SRUC, Edinburgh, United Kingdom

(2) DSM Nutritional Products, Venlo, Netherlands

(3) DSM Nutritional Products, Basel, Switzerland

(4) DSM Nutritional Products, Village neuf, France

* Corresponding author: jos.houdijk@sruc.ac.uk

Muramidase enzymes can hydrolyse peptidoglycan, the main structural polymer of bacterial cell walls. When released into the gut during natural bacterial turnover, peptidoglycan in bacterial cell debris may detriment gastrointestinal functionality. Indeed, supplementation of a novel microbial muramidase (Balancius™) has been shown to benefit growth performance and gastrointestinal functionality in broilers (1,2,3).

Here, we evaluated the effect of Balancius™ in practical diets, supplemented with phytase, xylanase and a coccidiostat on growth performance, litter quality and footpath dermatitis of male broilers. Control wheat-soya bean meal based diets (T1) were supplemented with muramidase at 25,000 (T2), 35,000 (T3) or 45,000 (T4) LSU(F)/kg feed. The four dietary treatments were allocated to 96 pens, each with ten day-old Ross 308 male broilers in a randomized block design (n=24), using a three phase feeding strategy with starter (d0-10; crumbs), grower (d10-21; pellets) and finisher (d21-35; pellets) diets. From d14 to d18, feed was removed for 4 h/d from 8 am to induce a mild gut health disturbance. Analysis of variance showed that muramidase supplementation increased final bird weight from 2685 to 2772, 2804 and 2848 g (s.e.d. 32 g; P<0.001) and average feed intake from 3746 to 3816, 3887 and 3913 g (s.e.d. 41 g; P<0.001) for T1, T2, T3 and T4, respectively. This resulted in an improved mortality corrected feed conversion from 1.420 to 1.401, 1.414 and 1.401, respectively (s.e.d. 0.005; P<0.001). These beneficial impacts on broiler production efficiency were achieved largely over the first 21 days, and most pronouncedly during the grower phase, when the mild gut health disturbance was induced. In this study, muramidase supplementation did not affect litter score, pH, dry matter or nitrogen content at d35, which averaged 2.1±0.03, 8.28±0.050, 56.9±1.91% and 1.9±0.05%, respectively. Footpad dermatitis incidence at d35 was low and did not differ between treatments. We concluded that in the presence of a mild gut challenge but otherwise good rearing conditions, dietary supplementation of Balancius™ on top of feed enzymes and a coccidiostat significantly improved broiler production efficiency during the starter and grower phase, which carried through to five weeks of age.

(1) Lichtenberg et al 2017. Regul Toxicol Pharmacol 89, 57-69

(2) Goodarzi Boroojeni et al 2019. Poult Sci 98, 2080-2086

(3) Sais et al 2019. Poult Sci in press; doi.org/10.3382/ps/pez466
ID: 1403

EVALUATION OF PARTICLE SIZE, FEED FORM AND PELLET DIAMETER ON BROILER PERFORMANCE AND
PROCESSING YIELD FROM 1 TO 39 DAYS OF AGE

M. Rueda lastres*(1), W.Pacheco(1), C.Starkey(1), J.Caldas-cuevas(3), J.Dunn-starkey(1)

Particle size and feed form manipulation have emerged as alternatives to improve poultry
performance. The objective of the study was to elucidate the effect of particle size and feed form on
broiler performance and processing yield from 1 to 39 d of age. A total of 1800 d old male Cobb 500
birds were randomly assigned to 9 dietary treatments with 8 replicate pens per treatment and 25 birds
per pen. The experiment consisted of a 3 × 3 factorial arrangement of 3 corn particle sizes (750, 1150
and 1550 µm) and 3 feed forms (mash, 3.0- and 4.0-mm pellets) provided from 1 to 39 d. Feed intake
and BW were determined at 17, 27 and 39 d of age and FCR was calculated by using the weights of
mortality. On d 40, 10 birds per pen were processed and on d 41, carcasses were deboned to determine
meat yield. Data were statistically evaluated as a 3 × 3 (feed form × corn particle size) factorial
arrangement in a randomized block design. Broilers fed 3.0- and 4.0-mm pellets had higher BW, FI and
lower FCR (P<0.05) compared to broilers fed mash at 39 d of age. Broilers fed diets with 750 µm corn
particle size had higher BW (P<0.05) than broilers fed diets with 1550 µm at 39 d of age, while broilers
fed diets with 1150 µm had an intermediate BW. Broilers fed diets with 750 µm corn particle size had
higher FI (P<0.05) than broilers fed diets with 1150 and 1550 µm at 39 d of age. Particle size of corn did
not influence FCR at 39 d of age. Broilers fed 3.0 mm pellets had the heaviest (P<0.05) carcass and
breast weights, followed by broilers fed 4.0 mm pellets and the lowest weights by broilers fed mash.
Tenders and wings weight were similar among broilers fed 3.0- and 4.0-mm pellets, but higher (P<0.05)
than broilers fed mash. Broilers fed diets with 750 µm corn particle size had heavier carcass and breast
weight (P<0.05) than broilers fed diets with 1550 µm, with 1150 µm having an intermediate carcass
and breast weight. Furthermore, broilers fed diets with 750 µm corn particle size had higher tender
weight and yield (P<0.05) than broilers fed diets with 1150 and 1550 µm. These results showed that
feed form and particle size influence broiler performance and processing yield of Cobb 500 broilers.

<footnote>(1) Auburn University, Auburn, United States</footnote>

<footnote>(3) Cobb, Siloam springs, United States</footnote>

<footnote>* Corresponding author: msr0036@auburn.edu</footnote>
EFFECTS OF CORN PARTICLE SIZE AND FORMIC ACID ON PRODUCTIVE AND PROCESSING PERFORMANCE OF BROILERS

A. Rubio\*(1), J.Pope(2), J.Jendza(3), A.Fahrenholz(1)

(1) North Carolina State University, Raleigh, United States

(2) House of Raeford Farms, Raleigh, United States

(3) BASF, Florham park, United States

* Corresponding author: aarubio@ncsu.edu

Previous research has reported that inclusion of coarse corn particle size in broiler diets can improve gastro-intestinal tract (GIT) function. Formic acid decreases the feed pH which could create a less favorable environment for bacteria and alter buffering capacity which may improve the efficacy of digestive enzymes. However, research evaluating the interactive effects between coarse corn particle size and dietary levels of formic acid throughout the production period of broilers is sparse. The objective of this study was to evaluate the effects of corn particle size and formic acid on broiler growth performance and carcass characteristics. A total of 2,034 male Ross YPM × 708 broiler chicks were randomly distributed among 6 dietary treatments with 12 replicate pens/treatment and 32 birds/pen, which constituted a 3 × 2 factorial arrangement of 3 corn particle sizes (395 µm fine corn (FC), 850 µm coarse corn (CC) obtained with greater inclusions of 1642 µm corn (CC-Hi) or lesser inclusions of 2290 µm corn (CC-Lo) when blended with FC and 2 dietary levels of formic acid (0.0% and 0.4%) fed from 11-49 d of age. Feed intake and BW were determined at 10, 21, 35, 42 and 48 d of age and feed conversion (FCR) was calculated and adjusted for mortality. At 43 d, 2 birds/pen were processed for the determination of carcass characteristics and hot carcasses were deboned to determine total breastmeat yield (pectoralis major and minor muscles). Data were statistically evaluated as a 3 × 2 (corn particle size × formic acid) factorial arrangement in a randomized complete block design. Although there were no significant differences in FI attributed to the inclusion of formic acid (P>0.05), birds fed CC-Hi diets had a lower FI compared with those fed CC-Lo diets at (P<0.05) at 48 d. At 35 d, birds fed FC diets with 0% formic acid had a lower BW compared with those fed FC diets with 0.4% formic acid. Birds fed CC and 0.4% formic acid exhibited an improved FCR compared with those fed diets with FC and 0% formic acid at 48 d. Dietary inclusion of formic acid increased pectoralis minor yield at 43 d (P<0.05). These data indicated that up to 0.4% of formic acid can be fed with a coarse corn particle size without negatively affecting broiler performance or carcass characteristics.
MINERAL NUTRITION
IMPACT OF AN E. COLI 6-PHYTASE ON IN VITRO PHYTATE BREAKDOWN OF DIFFERENT FEEDSTUFFS

E. Vanderbeke\*(1), L.Nollet(1), L.De jonge(2)

(1)Huvepharma NV, Antwerp, Belgium

(2) Wageningen University, Wageningen, Netherlands Antilles

(3)Azad University, Tehran, Iran

* Corresponding author: lode.nollet@huvepharma.com

The aim of this study was to estimate the effectivity of different levels of a E. coli 6 phytase (OptiPhos®, Huvepharma NV) on the in vitro degradation of phytate in six feed ingredients (soybean meal, rapeseed meal, sunflower seed meal, barley, maize, and wheat). The in vitro approach was based on an incubation which simulated the conditions in the stomach and the small intestine. The incubation was conducted in triplicate according to the following procedure: 0.8 g of feed ingredient and 20 ml of pepsin-HCl solution (pH 3.0), with or without phytase, was incubated during 1.5 hours at 37°C. After neutralisation, 20 ml acetate buffer (pH 5.5) was added and the mixture was incubated (2.5 h, 37°C). The incubation was stopped and the supernatant and residue was separated by centrifugation (4000g, 15 min). Samples of the supernatant were analysed on P while also the P and phytate-P content in the feed ingredients and the amount of P in the residues were analysed. Based on the amount of inorganic P into solution at the end of the trial of the phytase and non-phytase treated samples, it could be calculated how much P per gram of feed materials has been transferred into soluble inorganic P when supplementing different levels of OptiPhos®. This corresponds to the breakdown of mainly soluble phytate-P due to the action of OptiPhos®. Results indicated that adding OptiPhos® increased significantly the amount of total P release (mg per g feed material) strongly at 500 FTU/kg, while increasing the levels of OptiPhos® resulted in extra P release (p> 0.05). Of the three protein sources, the improvement (%) in phytate-P degradation vs the control was highest for sunflower seed meal (79, 81 and 86 %) and rapeseed meal (70, 69 and 70 %) compared to soybean meal (54, 56 and 59 %) at 500, 1000 and 1500 FTU/kg respectively. Of the three cereals, the phytate-P breakdown was highest in corn (70, 72 and 76 %) and less in barley (39, 46 and 49 %) and wheat (50, 52 and 52 %) at 500, 1000 and 1500 FTU/kg respectively. It could be concluded that OptiPhos® (a) led to a higher inorganic P release from phytate-P in raw materials with higher phytate-P levels, (b) had even a more efficient phytate breakdown (%) in feedstuffs containing high phytate-P levels (sunflower seed meal and rapeseed meal) than in feedstuff containing lower phytate-P levels and (c) that from the 3 cereals, phytate breakdown by OptiPhos® was higher in corn compared to barley and wheat.
DIETARY ZINC-L-SELENOMETHIONINE IMPACTS PROGENY OF BROILER BREEDER HENS

T. Noetzold*(1), R.Horn(1), P.Dos santos(1), A.Favero(1), A.Fireman(2), L.Kindlein(1), S.Vieira*(1)

(1)Federal University o f Rio Grande do Sul, Porto alegre, Brazil

(5)Zinpro Corporation, Eden prairie, United States

* Corresponding author: thiagonoetzold@gmail.com

Selenium is one the most controversial trace elements. Although toxic at high doses, deficiency is related to increased disease susceptibility of both animals and humans, and decreased productive and reproductive performance of farm animals. A study was conducted to evaluate the effect of inorganic Se, zinc-L-selenomethionine, and Fe supplementation on broiler breeder hen performance. A total of 132 Cobb 500 broiler breeder hens at 20-weeks-of-age (woa) were placed in individual cages. Hens were randomly assigned to 11 treatments of Se and Fe trace mineral supplementation: Control with 55 ppm Fe as FeSO4 + 0.30 ppm sodium selenite (Se-S; T1). Treatments 2 through 9 (T2-T9) included Se-S at 0.30 ppm and increasing levels of Se as zinc-L-selenomethionine (ZnSeMet) at 0.15, 0.20, 0.30,and 0.40 ppm, with either FeSO4 at 55 ppm or Fe amino acid complex (Fe-AA) at 40 ppm. Treatment 10 included 0.30 ppm Se as ZnSeMet + 55 ppm Fe as FeSO4, and T11 had 0.30 ppm Se as ZnSeMet + 40 ppm Fe as Fe-AA. Hens were adapted from 20 to 26 woa. Data collection was completed in 3 periodsof 28 days each, from 27 to 38 woa, with one incubation per period. All data were analyzed using proc MIXED of SAS, and means compared using the Tukey-Kramer test, where differences were considered significant at P < 0.05. Dietary treatment did not impact overall hen productive performance or incubation, except for hatchability of settable eggs, which was higher in T11 compared with Control (P < 0.05). In T3, T4, T5, and T7, chick length was improved over the Control (P < 0.05). Groups with higher Se generated lower chick leg score abnormalities than groups fed lower Se in partial replacement (P < 0.05). Selenoprotein-W (SelW), which can serve as an antioxidant in different cell types, did not differ among treatments at d 14 of embryo incubation development (E14; P > 0.05), nevertheless it had a significant difference at E18 and E21 (P < 0.0001), where SelW gene expression decreased with increasing Se levels compared to Control (P < 0.05), except for T6 at E18. This result may indicate that SelW could be compensated due to higher Se available provided by breeder diets. In conclusion, ZnSeMet supplementation promotes improvement in hatchability of settable eggs, chick length and score, and lower SelW expression.

Keywords: broiler breeder, chick, leg score, selenoprotein-W.
PARTIAL AND TOTAL REPLACEMENT OF INORGANIC TRACE MINERALS BY AMINO ACID COMPLEXED MINERALS IN DIETS OF BROILER BREEDER HENS

T. Noetzold\*(1), G. Martins(2), C. Fiorenzano(2), C. Silva(2), A. Fireman(3), L. Kindlein(2), S. Vieira\*(2)

(1) Federal University of Rio Grande do Sul, Porto alegre, Brazil

(2) Federal university of Rio Grande do Sul, Porto alegre, Brazil

(3) Zinpro Corporation, Eden prairie, United States

\* Corresponding author: thiagonoetzold@gmail.com

Feeding organic trace minerals (OTM) to broiler breeders is common practice in industry, however, totally replacing inorganic trace minerals (ITM) with reduced OTM levels is innovative. This study evaluated the effect of partial and total replacement of inorganic TMs (sulfates of Zn, Mn, Cu, Fe and Se) with amino acid complexed minerals (AACM) in broiler breeder diets. Eighty-four Cobb 500 broiler breeder hens at 20 wks of age (woa) were placed in individual cages, with treatments provided from 27 to 38 woa after a 6-wk adaption period. Experimental diets were composed of 7 treatments: ITM Full (Zn: 100/0, Mn: 100/0, Cu: 20/0, Fe: 60/0, Se: 0.30/0, and I: 2/0 ppm); ITM Reduced (Zn: 60/0, Mn: 60/0, Cu: 13/0, Fe: 40/0, Se: 0.15/0, and I: 2/0 ppm); ITM Plus (Zn: 120/0, Mn: 120/0, Cu: 30/0, Fe: 70/0, Se: 0.40/0, and I: 3/0 ppm); AACM Partial Replacement (Zn: 60/40, Mn: 60/40, Cu: 13/7, Fe: 40/20, Se: 0.2/0.15, and I: 0/2 ppm); AACM Plus Replacement (Zn: 60/60, Mn: 60/60, Cu: 13/12, Fe: 30/40, Se: 0.2/0.2, and I: 0/3 ppm); AACM Reduced (Zn: 0/40, Mn: 0/40, Cu: 0/7, Fe: 0/20, Se: 0/0.15, and I: 0/2 ppm); and AACM Extra (Zn: 0/60, Mn: 0/60, Cu: 0/12, Fe: 0/40, Se: 0/0.2, and I: 0/3 ppm). All data were analyzed using proc MIXED of SAS and means compared using the Tukey-Kramer test, with differences considered significant at P < 0.05. Dietary treatments did not impact hen productive performance. Eggshell thickness increased with AACM supplementation (P < 0.05), and eggshell breaking strength was higher in total replacement compared with ITM treatments (P = 0.055). Egg weight prior to incubation was lower for partial replacement treatments, compared to ITM sources and total replacement treatments (P < 0.05). These differences were maintained for chick weight. Chick length was higher in partial replacement treatments of AACM, compared to ITM forms (P < 0.05). Percentage of leg score 1 (normal legs/toes), and 2 (signs of inflammation/redness in the legs) were higher and lower, respectively, in chicks from hens fed with total replacement treatments of AACM sources compared to ITM sources (P < 0.05), showing less leg abnormalities. There were no differences in navel button score of hatchings among treatments. In conclusion, diets supplemented with AACM led to improvements in eggshell thickness and shell breaking strength, along with hatching chick length and leg score, indicating better chick quality.

Keywords: broiler breeder, leg score, eggshell thickness, trace mineral
GROWTH PERFORMANCE RESPONSES OF BROILER CHICKENS TO PHYTATE PHOSPHORUS AND PHYTASE CONCENTRATIONS DURING DAYS 1 TO 11 OR 12 TO 23 POST HATCHING

O. Babatunde\*(1), A.Bello(2), Y.Dersjant-li(2), O.Adeola(1)

(1) Purdue University, West Lafayette, United States

(2) Dupont Nutrition and Biosciences, Leiden, Netherlands

* Corresponding author: bolufemi@purdue.edu

Common feed ingredients for broiler chickens have phosphorus (P) bound in the unavailable form of phytate complexes. However, phytase enzyme releases some of this P necessary for growth in broiler chickens. This study evaluated the effect of phytate P (PP) and phytase concentrations on broiler chickens during days 1 to 11 (phase 1) or 12 to 23 (phase 2) post hatching using growth performance as response criteria. Adequate P positive control (PC) diets in phase 1 or 2 contained 4.4 or 3.9 g non-phytate P (nPP)/kg, respectively; low-P negative control (NC) diets in phase 1 or 2 contained 2.3 or 1.7 g nPP/kg, respectively (without inorganic P, but with added meat & bone meal). Broiler chicks were randomly assigned to 6 replicate cages in a 3 × 5 + 1 factorial arrangement of treatments with 3 low-PP NC [2.3, 2.8, or 3.3 g PP/kg], 5 phytase levels (0, 500, 1,000, 2,000, or 4,000 FTU/kg) and a P-adequate PC. The PP was from raw feed materials while the phytase is a next generation biosynthetic bacterial 6-phytase. In phases 1 and 2 respectively, 1,152 and 768 male broiler chicks were used. In both phases, birds fed the NC had lower (P
DOSE RESPONSE TO AVAILABLE PHOSPHORUS IN TWO STRAINS OF AGED WHITE LAYING HENS

L. Lorieau*(1), B.Cheve(1), B.Renouf(1)

(1)Euronutrition, Saint-symphorien, France

* Corresponding author: lucie.lorieau@euronutrition.fr

The eggshell quality continues to be a major concern of the egg industry. Despite the numerous studies on phosphorus (P) requirements in the laying hen, the optimum dietary level of P remains unknown in aged laying hens. Unfortunately, P is an expensive nutrient, which might have adverse consequences on the environment. Moreover, requirements are affected by genetic which is also constantly changing. Consequently, a trial was performed to evaluate P requirements in two white hens strains.

At 76 weeks of age, 480 hens of each strain (A and B) were allocated in 5 groups (4 or 5 replicates of 20 animals), consisting of increasing levels of available P (0.13, 0.20, 0.27, 0.34 and 0.41%) and the same level of calcium (3.80%), for 16 weeks. The performance (egg production, weight of hens and feed consumption) and quality of the eggs were recorded weekly. Two-way ANOVA was performed to compare strains and P levels. Broken-line model was used to find the optimum P level.

The results showed that the increase of the available P concentration led to an increase of the feed consumption from 101.5 to 107.1 g on average for both strains (P <0.05) and limited weights loss from -140 g to -55 g (P <0.05) between 76 to 92 weeks of age. Following these results, the optimal dose of available P using broken-line analysis was between 0.27 and 0.33%.

In terms of strain, the feed efficiency was improved by 0.13 point for the strain A. Consequently, at 92 weeks, hens of the strain A weighed approximately 83 g more than strain B (P<0.001). There was no significant difference between the two strains for feed consumption or rate of lay. In terms of egg quality, the egg mass was greater for strain A compared with strain B (A : 66.7 g vs B : 61.9 g, P<0.001).

However, eggs of strain A were more downgraded (+ 5.1 points; P < 0.001) with more broken and dirty eggs. Moreover, the eggs of the strain B had a greater shell static stiffness: +3.5 N / mm (P<0.05).

Also, no statistical difference was observed for the eggshell index, eggshell breaking strength, bone mineralization of the tibias, regardless the strains and the levels of available P.

Thus, despite an extended range of P, the quality of the eggshell was only slightly improved by increased of P intake. However, the strain was preponderant on the egg quality. This type of dose-response trial is a good tool for studying the impact of a nutrient on the performance of laying hens.
EFFECT OF DIFFERENT TRACE MINERAL PROGRAMS ON PERFORMANCE OF BROILER BREEDER GRANDPARENTS

L. Linares*(1), C.Torres(1), F.Defontaine(1), R.Bouvet(2), C.Alleno(2)

(1)Zinpro Corporation, Eden prairie, United States

(2) Zootests, Ploufragan, France

* Corresponding author: linares@zinpro.com

Several organic trace mineral (TM) forms have been developed as supplements to animal feed. The type of chelate, such as amino acid (AA) or organic acid, and its binding properties are key factors differentiating organic TMs and effects in livestock performance are expected. Minerals complexed to AA (AACM) is the result of a soluble metal salt chelation to a single AA; hence, being absorbed via AA pathway. This study investigated the effects of supplementing zinc (Zn), manganese (Mn), copper (Cu), and iron (Fe) to broiler breeders as inorganic sulfates, chelate of methionine hydroxy analog (MHA), or as AACM on hen performance. A total of 1080 Hubbard A-7 Line D grandparent (GP) hens were assigned to 1 of 3 treatments: T1) Control, sulfates of Zn (80 ppm), Mn (80 ppm), Cu (17 ppm), Fe (40 ppm); T2) MHA, Zn (50 ppm), Mn (50 ppm), Cu (10 ppm), Fe (65 ppm, as FeSO4); T3) AACM, AACM of Zn (40 ppm), Mn (40 ppm), Cu (7 ppm), Fe (40 ppm) + sulfates of Zn (40 ppm), Mn (40 ppm), Cu (10 ppm). All diets had equal methionine supplementation levels. Each treatment had 15 repetitions, and each replicate contained 6 cages with 4 hens per cage. Females and males were fed the same experimental diets from 35- to 59-weeks-of-age (woa). Data were analyzed using ANOVA and c2 tests. The AACM breeders had greatest egg quality (Haugh unit and albumen height at 40 woa, $P < 0.006$), followed by MHA or Control hens. Likewise, hatchability of eggs set at 40 woa tended to improve ($P = 0.13$) and late embryo mortality [$>14$ day of incubation (doi)] tended to be reduced ($P = 0.10$) with AACM supplementation, followed by MHA and Control. Early embryonic mortality ($P = 0.086$). A total of 500 grade-A male chicks from each treatment at the 59 woa hatching were placed in floor pens (10 pens/treatment × 50 chicks/pen), and body weight and mortality rate were measured at d 7. Progeny from hens fed organic TM had greatest body weights in comparison to Control ($P = 0.004$). In conclusion, the feeding program with a partial replacement of sulfates by AACM improved egg quality and reproductive performance of Hubbard A-7 Line D GP broiler breeders, with positive carry-over effects in progeny during the first week of growth.
ADDRESSING ENVIRONMENTALLY SUSTAINABLE POULTRY PRODUCTION WITH REDUCED COPPER SUPPLEMENTATION IN BROILERS REARED UNDER 'NON-ANTIBIOTIC EVER' CONDITIONS CHALLENGED WITH CLOSTRIDIUM PERFRINGENS

D. Neves*(1), M.Rebollo(1), L.Linares(1), D.Moore(3)

Environmentally sustainable poultry production starts with a reduction in excess environmental nutrient excretion, while maintaining flock performance and health status. Although diseases such as necrotic enteritis (NE) undermine this strategy, they also intensify the search for more efficacious nutrients to address them. For this, two experiments were carried out with the objective of evaluating the effect of reduced copper (Cu) supplementation on performance and Cu excretion in broilers reared under a 'non-antibiotic ever' (NAE) system. In Experiment 1 (Exp1), 1,296 one-day-old Cobb 500 chicks, vaccinated for coccidiosis at hatch, were randomly assigned to 1 of 3 treatments: 1) 125 ppm Cu as tribasic copper chloride (TBCC); 2) 21 ppm Cu as copper amino acid complex (AAC); and 3) 21 ppm Cu as AAC in a step-down program (AAC SD). In Experiment 2 (Exp2), 1,728 chicks were vaccinated against coccidiosis at hatch, followed by an in-feed challenge with Clostridium perfringens (CL-15, Type A strain, 1X109/bird) at 17 days-of-age, were randomly assigned to 1 of 4 treatments: 1) 125 ppm Cu as TBCC (TBCC-Cp); 2) 21 ppm Cu as AAC (AAC-Cp); 3) 21 ppm Cu as AAC in a step-down program (AAC SD-Cp); and 4) 14 ppm Cu as AAC in a step-down program (AAC LowSD-Cp). Data obtained was analyzed by two-way ANOVA, using the GLM procedure of SAS. Means were separated by the Tukey test when P ≤ 0.05. In Exp1, there were no significant differences in performance or mortality at d 42, nor in footpad scores at d 35. There was significantly lower excretion of Cu for both AAC and AAC SD treatments at 18 days (P < 0.0001), and for AAC at 42 days (P < 0.0001) compared to TBCC. Liver Cu was higher in treatments supplemented with AAC at both 21 (P < 0.06) and 35 days (P < 0.0001) compared to TBCC. In Exp2, there were no differences in performance or NE mortality at 42 days. At days 21, 28, and 35, intestinal lesion scores presented no differences between treatments. At d 35, treatments also showed no differences in footpad scores. In summary, there were no relevant differences in performance and health status of birds in either of the studies when Cu supplementation was reduced. This data suggests that decreasing overall Cu supplementation through the use of AAC Cu can maintain production performance while reducing Cu excretion and improving environmental sustainability.

<footnote>(1) Zinpro Corporation, Eden prairie, United States</footnote>

<footnote>(3) Colorado Quality Research, Wellington, United States</footnote>

<footnote>* Corresponding author: dneves@zinpro.com</footnote>
ZINC AMINO ACID COMPLEX IMPROVES INTESTINAL HEALTH PARAMETERS IN BROILERS REARED
UNDER CHRONIC CYCLIC HEAT STRESS CONDITIONS

A. De grande*(1), E.Delezie(2), C.Rapp(3), C.Torres(3), R.Ducatelle(1), S.Leleu(2), F.Van immerseel(1)

(1)Ghent University, Merelbeke, Belgium

(2) Research Institute for Agriculture, Fisheries and Food (ILVO), Animal Sciences Unit, Merelbeke, Belgium

(3) Zinpro Corporation, Boxmeer, Netherlands

* Corresponding author: annatachja.degrande@ilvo.vlaanderen.be

Heat stress (HS) not only impairs animal health and performance, but also negatively impacts intestinal
morphology and gut barrier integrity in broilers. The objective of this study was to evaluate the effect
of supplementing zinc (Zn) as ZnSO4 or Zn amino acid complex (ZnAA) on intestinal health in broilers
subjected to chronic cyclic HS (32°C ± 2°C and relative humidity of 50 to 65% for 6 h daily) applied in
the finisher period. It has been suggested that vitamin E (Vit E) may help alleviate negative effects of
HS, and therefore an increased inclusion level of Vit E was used as a positive control (100 IU/kg). A total
of 1224 day-old male Ross 308 broilers were randomly distributed among 4 dietary treatments (9
replicates of 34 birds per treatment), organized in a 2x2 factorial arrangement: two Zn sources (60 ppm
Zn as either ZnSO4 or ZnAA) combined with two levels of Vit E (50 or 100 IU/kg) were added to a
wheat/rye-based diet. A 3-phase feeding scheme was applied: Starter (d 0 to 10), Grower (d 10 to 28)
and Finisher (d 28 to 36). Intestinal health was evaluated by measuring villus length (VL) and crypt
depth (CD), along with infiltration of CD3+ T-lymphocytes in the propria mucosae in duodenum
sections, and ovotransferrin leakage in ileum content. Statistical analysis was performed using R for
Windows (v3.5.1) utilizing a General Linear Model (GLM)) with ‘Zn source’ and ‘Vitamin E level’ as fixed
factors and block as a random factor (factorial analysis; pen as experimental unit). A main effect of zinc
source on intestinal health parameters was observed, however, no interactions or main effect of Vit E
was observed. Supplementation with ZnAA, independent of Vit E level, increased VL (d 10: +16%, P <
0.001; d 28: +15%, P = 0.028; d 36: +7%, P = 0.009). Likewise, at the end of the Finisher phase (d 36),
infiltration of CD3+ T-cells in duodenum was decreased (11.2 vs 13.1 area %, P = 0.023) and
ovotransferrin leakage in ileum content was lower (15.99 vs 20.12 µg/g, P = 0.094) in broilers fed ZnAA,
independent of Vit E level. In conclusion, replacing ZnSO4 with ZnAA in diets of broilers subjected to
heat stress improved intestinal morphology and mitigated intestinal inflammation.
THE EFFECT OF LIMESTONE SOLUBILITY AND PHYTASE DOSE ON PHOSPHORUS DIGESTIBILITY IN BROILERS

A. Debicki-garnier\*(1), M.Sinclair-black(2), P.Plumstead(2), Y.Dersjant-li(3)

(1)Danisco Animal Nutrition (IFF), Paris, France

(2)Chemunique & Department Wildlife and Animal Sciences, University of Pretoria, Lanseria, South Africa

(3) Danisco Animal Nutrition (IFF), Oegstgeest, Netherlands

* Corresponding author: Anne-Marie.Debicki-Garnier@iff.com

This study evaluated the effect of two limestones from a single source, with high and low solubilities and phytase doses on phosphorus (P) digestibility in broilers. Male Ross 308 broilers were reared in floor pens on starter (0-10d) and grower diets (11 to 19 or 23d) formulated to breed recommendations. At 19d and 23d, broilers were weighed individually, and allocated to 80 battery cages, with 6 birds per cage, based on similar BW per cage. The trial design was a 2x5 factorial arrangement with 2 limestone particle sizes/solubilities (0.123 and 0.722 mm geometric mean diameter (GMD) with 96 and 26% solubility at 5 min respectively), and 5 dose levels (0, 250, 500, 1000 and 2000 FTU/kg) of Buttiauxella sp. phytase expressed in T. reesei. The 10 test diets were randomly assigned to 8 replicate cages and fed for 34h in two blocks from 19-21d or 23-25d. Diets were based on corn, soybean meal, full-fat soya, canola, and sunflower oil cake and contained 0.4% Cr2O3 as a marker, fed ad lib in mash form. Limestone Ca was analyzed and added to the basal diet that contained no added inorganic P to achieve 0.75% Ca in test diets. After 34h of feeding test diets, the distal half of the ileum digesta was collected for apparent ileal digestibility (AID) of P measurement. Data were analysed as a two-way ANOVA using fit model in JMP (JMP 14) at a significance level of P < 0.05. No interaction was found between limestone solubility and phytase dose. Low solubility limestone increased AID P. Increasing phytase dose resulted in step wise increase (P<0.05) in AID P with each incremental phytase dose. The AID P increased with increasing phytase dose from 0 to 2000 FTU/kg from 21.2 to 75.1%, and 35.5 to 83.5% with high and low soluble limestone respectively. Phytase at 2000 FTU/kg released 1.97 (0.87 to 3.08) and 2.21 (1.46 to 3.42) g/kg digestible P, respectively for low and high soluble limestones, calculated as digestible P above the non-supplemented diets. The measured digestible P content in NC was lower than the calculated value (1.49 g/kg, CVB) with high soluble limestone, which may lead to risk of P deficiency. These data confirm previous findings that a rapidly soluble fine limestone results in lower P digestibility in broilers. Increasing phytase to 2000 FTU/kg resulted in incremental steps in AID P and can be used as a strategy to reduce negative effects of limestone on P utilization and thereby improve the environmental sustainability of broiler production.
EFFECTS OF PHYTASE SUPPLEMENTATION ON PERFORMANCE, EGG QUALITY AND BLOOD PARAMETERS IN LAYING HENS FED ON DIFFERENT LEVELS OF NONPHYTATE PHOSPHORUS

K. Ryu*(1), C.Lim(2), M.Rana(2), H.Choe(3)

(1)Department of Animal Science, Jeonbuk National University, Jeonju, South Korea

(2)Jeonbuk National University, Jeonju, South Korea

(3)Department of Animal Biotechnology, Jeonbuk National University, Jeonju, South Korea

* Corresponding author: seon@jbnu.ac.kr

An experiment was conducted to investigate the effects of various levels of nonphytate phosphorus with phytase supplementation on performance, egg quality, and some blood biochemical parameters in laying hens. A total of five hundred and forty 61 week-old Hy-line Brown laying hens were randomly allotted to six dietary treatments with five replicates and eighteen hens in each replicate. Three isocaloric and isonitrogenous diets were formulated to contain 2.0, 2.5 and 3.0 g NPP/kg diet with the level of 45 g Cakg-1 diet and supplemented with or without phytase 1000 FTY per kg diet (Ronozyme HiPhos-L, Aspergillus oryzae 6-phytase). All laying hens were subjected to the same management practices during the experimental period. Productive performance and egg quality were measured every four weeks for the entire eight-week experiment from 61 to 68 weeks of age. The Serum biochemical parameters were assessed at the end of the experiment. Data were analyzed in a completely randomized design using the GLM procedure of SAS software (SAS 9.1, 2009). The results showed that no significant differences were seen in hen-day egg production, feed intake, egg weight, egg mass and feed to egg mass ratio of laying hens at 64 and 68 weeks of age, however, egg production was tended to be numerically higher in phytase supplemented groups at 68 weeks of age as compared to the without phytase added groups. Egg quality including albumen height, Haugh unit, yolk color, shell color and egg shell breaking strength were not influenced by dietary treatments whereas egg shell thickness was significantly higher in phytase supplemented groups than that of unsupplemented at 68 weeks of age. Moreover, there was no influence effect of dietary treatments on albumin, total protein, glucose, cholesterol, high-density lipoprotein, aspartate aminotransferase, alanine aminotransferase and triglyceride concentrations in serum. In conclusion, dietary nonphytate phosphorus with supplemented phytase can improve egg production and egg shell thickness in laying hens.
CUO NANOPARTICLES INFLUENCE ON THE QUAILS EGG PARAMETERS AND GROWTH PERFORMANCE

V. Slausgalvis*(1), L.Tribulas(2), V.Semaška(3), R.Bobinienė(2), D.Gudavičiūtė(2), D.Vencius(2), A.Gefenienė(2)

(1) Central Park, JSC, Kaunas, Lithuania

(2) Vytautas Magnus University. Education Academy, Vilnius, Lithuania

(3) WPSA Lithuanian Branch, Vilnius, Lithuania

* Corresponding author: diana.gudaviciute@vdu.lt

It has been becoming increasingly important to produce micronutrients of the size of nanometers. The aim of the research was to synthesize a nanoform of a trace mineral feed additive. The nanoform of trace mineral feed additives could be applied in poultry to improve the absorption of trace minerals and to decrease the amount of trace minerals in premixes. The research was carried out in order to investigate the influence of CuO nanoparticles on growth performance and egg parameters of Japanese quails (Coturnix coturnix japonica).

CuO nanoparticles were synthesized applying a wet chemical sol-gel method. For the synthesis of CuO, CuSO4 • 5H2O, NaOH and citric acid were used. Cu content in eggs was determined using the inductively coupled plasma emission spectrometry. The quails were grown in cages from the first day of age to 12 weeks of age. For the trial, the quails were divided into three groups. The first group was a control group, and the quails were fed with standard feed. The feed for the quails in the second group was enriched with the inorganic CuSO4 • 5H2O additive. The feed for the quails in the third group was enriched with the CuO nanoparticles additive. The following parameters were investigated: the amount of Cu in eggs yolk and albumen, the influence of the CuO nanoparticles additive on the growth of quails, laying intensity, and an average egg weight.

The experiment and the analysis of the study showed: the highest level of Cu in the egg albumen was observed in the quails at 12 weeks of age in the second trial group (2.941 µg/g), i.e. it increased by 25.74 percent in comparison with the control group (p increased from 206.73 g to 218.12 g, i.e. it increased by 5.51 percent in comparison with the control group (p

Keywords: nanoparticles, copper, trace element, egg parameters, quails.
PERFORMANCE AND MINERALIZATION IN BROILERS: WHAT ARE THE BEST DIETARY LEVELS OF AVAILABLE PHOSPHORUS AND CALCIUM, REGARDING CALCIUM TO AVAILABLE PHOSPHORUS RATIO?

M. Bournazel(1), S.Klein(1), A.Mathiaud*(1), P.Moquet(1)

(1)MIXSCIENCE, Bruz, France

* Corresponding author: adeline.mathiaud@mixscience.eu

Low phosphorus (P) diets may support sustainability in broiler production. In fast growing broilers, it is, however, observed that low P diets leads to inconsistent bone mineralization and welfare outcomes. This may be due to differences in available P (aP) utilization. The latter is conditioned by dietary calcium (Ca) supply. Currently, little is known about aP and Ca:aP requirements of broilers. Thus, a trial was performed to study the effect of dietary aP and Ca:aP ratio on the growth performance, carcass yield and tibia characteristics. A total of 384 day old male Ross 308 broiler chicks were randomly assigned to six dietary groups, each replicated 16 times, for 42 days (d). Dietary groups consisted in different feeding sequences with low (L), medium (M) and high (H) aP levels in the starter (L:0.45; M:0.50, H:0.55), grower (L:0.35, M:0.40, H:0.45) and finisher feeds (L:0.27, M:0.32, H:0.37) with low (1.8) or high (2.2) Ca:aP ratios. Groups were arranged as follows: LLL2.2, MMM2.2, HHH2.2, HHL2.2, LLL1.8 and HHH1.8. Orthogonal contrasts were performed for statistical analysis. Dietary treatments did not differ significantly in growth performance during the starter and grower phase. In the finisher phase, FCR and ADG tended to improve linearly with aP level at the highest Ca:P ratio (P<0.1). A decrease in growth performance was observed with low aP in finisher phase, suggesting a high aP requirement during this phase. Concerning bone characteristics, no effect was observed at 21 d. At 42 d, the decrease of aP in HHL2.2 led to lower tibial breaking strength (TBS) and stiffness (TS), resulting in value similar to LLL2.2. In contrast, HHH2.2 resulted in a higher tibial TBS and TS compared to HHL2.2 and LLL2.2 (P<0.05). LLL1.8 resulted in the lowest tibial TBS and TS values of all groups. HHH1.8 did not differ from HHH2.2 for those parameters. Concerning carcass traits, LLL2.2 had a significantly higher breast meat yield than LLL1.8 (+1.1 pt; P<0.05). In conclusion, this trial demonstrates that decreasing aP in the finisher phase negatively impacts growth performance. Low aP with low Ca:aP ratio seems to be the worst sequence. In contrast, high aP seems to be mandatory to achieve the best bone strength, irrespective of Ca supply. Overall, high aP with high Ca seemed to be the best strategy for both criteria. Future trials could investigate the effect of reducing aP and Ca during the starter and grower phases, e.g. with LLH2.2 or LLH1.8 sequences.
EFFECT OF A SODIUM ELECTROLYTIC ACIDIFIER ON LAYING HENS PERFORMANCE AND EGG SHELL
QUALITY

B. Iglesias*(1), M.Charriere(2), J.Azcona(3), G.Vicente(4)

(1)INTA, Pergamino, Argentina

(2) Sección Avicultura, EEA Pergamino, Instituto Nacional de Tecnología Agropecuaria (INTA), Pergamino, Argentina

(3) Granja Avícola "El Chaveche", Pergamino, Argentina

(4) Porfenc SRL, Ciudad autónoma de buenos aires, Argentina

* Corresponding author: iglesias.bernardo@inta.gob.ar

The dietary inclusion of acidifiers can contribute to the gut health and improve the absorption of some
minerals and others nutrients by a decrease in pH and a stimulation of the villus height. The aim of this
study was to evaluate the effect of the dietary addition of a sodium electrolytic acidifier on the
performance of laying hens under typical production conditions. The trial was carried out at the
facilities of a commercial egg producer to compare two treatments (T1: Control and T2: Idem to T1 plus
acidifier) during 4 periods of 28 days. Two repetitions per treatment were used. A total of 4202 laying
hens, were allocated in two barns (500 cm2 of cage per hen), one with 1868 Hy-Line W-80 (81 weeks
old) and another with 2334 Lohmann LSL-Lite (47 weeks old). A completely randomized block design
was used. Each barn was considered as a block and divided longitudinally conforming two experimental
units (lots). Control diets based on corn and soybean meal were formulated according to the lines
recommendations and hens feed intake. The same diets were used in T2 replacing part of sodium
chloride and corn by the acidifier (1.5 kg/MT). Each kg of acidifier replaced 600 g of sodium chloride.
The rest of nutrients were the same for both treatments. The acidifier used is based on a mixof natural
extracts (polyphenols from Schinopsis sp. and citrus pectins) and sodium bisulfate (AES®, Porfenc SRL).
Egg production (%), feed intake (g/day), egg weight (g), feed conversion ratio (g/dozen eggs) and egg
shell quality (mg/cm2) were analyzed at the end of each period, and body weight at theend of the 4th
period. Data were analyzed by a 2-way ANOVA. No differences in feed intake and egg weight were
observed between treatments throughout the four experimental periods (P>0.05), while egg
production was higher along all the trial when AES was used (Control: 78.2% vs. AES: 80.8%, P<0.05;
means of the 4 periods), so feed conversion ratio was also better with AES (Control: 2.00 kg of
feed/dozen eggs vs. AES: 1.92, P<0.10). Hens from Control treatment were heavier than those from
the AES treatment (Control: 1679 g vs. AES: 1634 g; P<0.10). On the other hand an improvement in egg
shell quality was observed (Control: 82.1 mg/cm2 vs. AES: 83.9 mg/cm2; P<0.10). This result was
associated with a reduction in the excreta calcium content. In conclusion, the inclusion of AES in laying
hens diets improved the egg production, feed conversion ratio and the egg shell quality.
OPTIMIZE PHYTASE DOSE BASED ON PHYTATE LEVELS IN BROILER FEED FORMULATION

C. Evans(1), Y.Dersjant-li(2), R.Hardy(1), A.Mereu*(2), L.Marchal(2)

(1)DuPont Animal Nutrition, Marlborough, United Kingdom

(2)DuPont Animal Nutrition, Leiden, Netherlands

* Corresponding author: ceinwen.evans@iff.com

Phytic acid [inositol (1,2,3,4,5,6) hexakisphosphate] or phytate (salt of phytic acid) is the major phosphorus (P) storage in plant-based ingredients. A large portion (60-80%) of the phosphorus in the plant-based ingredients occurs in the form of phytate. Mono-gastric animals have limited capability of utilizing the phytate-bound P. To improve the availability of plant-based phosphorus, phytase is commonly applied in poultry diets. When first launched, phytase was commonly used at 500 FTU/kg with the main purpose of reducing the P excretion to the environment. Recently, with more efficient and cost effective phytases in the market the phytase inclusion rate has increased to up to 5000 FTU/kg based on literature. However, optimal dose of phytase is related to the level of the phytate substrate in the diets. Phytate content of major feed ingredients could be derived from literature or feed tables but this does not account for variability in different plant-based feedstuffs, not only due to plant type, but also depending on the growing conditions and soil types and contents. This study reported the analyzed phytate-P content in 37 major feed ingredients sourced globally and analyzed by DuPont lab in the last 10 years. IP6 levels were determined by HPLC and then Phytate P calculated. In total more than 1000 samples were analyzed from more than 40 countries. The average Phytic acid, Phytate P content, variation, minimum and maximum levels were determined. The data showed there was a large variation in phytate content for the same feed ingredient, such as corn, the phytate-P content isin a range of 0.14 to 0.30% with a mean value of (256 samples). For SBM, phytate-P content is in a range of 0.28 to 0.54% (214 samples), and the range for wheat (97 samples) and barley (64 samples) is from 0.18 to 0.31% and 0.14 to 0.34% respectively. For ingredients with high phytate content, the variation is even bigger, for example, phytate-P content is in the range of 0.58 to 1.10% and 1.34 to 2.15% for wheat bran (61 samples) and rice bran (40 samples) respectively. Understanding the substrate and phytase ratio is important to determine the optimal phytase dose in feed formulation. Previous research has demonstrated that substrate level will impact the magnitude of response seen with phytase supplementation in terms of the extra-phosphoric effects (energy and amino acid sparing) and therefore knowing the accurate phytate level can be used to identify the optimal dose forfeed cost savings for a given diet.
EFFECT OF DIFFERENT SOURCES OF CU ON ANTIMICROBIAL RESISTANCE IN BROILER CHICKENS

A. Forouzandeh\*(1), L.Blavi(1), A.Vidal(2), M.Rodríguez(3), A.Monteiro(4), J.Pérez(1), L.Darwich(2), D.Solà-oriol(1)

(1)Department of Animal and Food Sciences, Universitat Autònoma de Barcelona, Bellaterra, Spain

(2)Department of Animal Health and Anatomy, Universitat Autònoma de Barcelona, Bellaterra, Spain

(3)PigCHAMP Pro Europa, Segovia, Spain

(4)Animine, Annecy, France

* Corresponding author: asal.forouzandeh@gmail.com

Copper (Cu) requirement for broilers is 8 mg/kg diet. The use of higher doses (125 to 250 mg/kg) has a growth promoter effect but may increase the antimicrobial resistance in broilers. Therefore, the objective of this trial (part of Project E! 11780) was to study the effect of 150 mg/kg of Cu as Cu sulfate (CuSO4), or as dicopper oxide (Cu2O, CoRouge®) on the antimicrobial resistance in Enterococcus spp. and Escherichia coli (E. coli). A total of 300 one-day-old male broilers (Ross 308) were distributed into 3 experimental treatments (10 pens/ treatment and 10 birds/ pen): negative control (NC), with no Cu addition, and NC with 150 mg/kg of Cu supplemented as CuSO4 or Cu2O. Broilers were challenged by reusing an old litter with high concentrations of Clostridium spp. At d 16, 28 and 42 fecal samples were collected from one broiler per pen. Samples were analyzed for microbiological identification of Enterococcus spp. and E. coli. A phenotypical antimicrobial resistance test was performed to determine the resistance of Enterococcus spp. and E. coli isolates against thirteen antimicrobial agents. Additionally, extracted bacterial DNA was used to detect the antimicrobial resistance genes of vancomycin (vanC1 and vanC2), tetracycline (tet M), erythromycin (erm B), copper (tcrB) and zinc (czcA). Data were analyzed using the chi-squared test. There were no differences among treatments on the phenotypic antimicrobial resistance on d 16 and 28. However, on d 42 the isolated Enterococcus spp. from broilers fed the NC diet had higher (P < 0.05) resistance to enrofloxacin, gentamicin, and chloramphenicol compared to isolates from birds fed 150 mg/kg of Cu as CuSO4 and Cu2O. By contrast, in the isolated E. coli from broilers fed 150 mg/kg of Cu as CuSO4 and Cu2O had higher (P < 0.05) resistance to streptomycin (78 and 56%, respectively) and chloramphenicol (56% on average) compared to isolates from birds fed the NC diet (11 and 0%, respectively). The percentage of resistant strains in Enterococcus spp. was high (> 80%) for all the studied genes, treatments, and days. However, in the isolated E. coli the frequency of resistant strains was lower (< 20%) for VanC1 and VanC2 in all treatments and days, but for TcrB gene resistance increased over time in all treatments. To conclude, the antimicrobial resistance is affected by time and by bacterial species and likely by the microbiota background, but the effect of Cu source remains unclear yet, and more research is required.
ID: 1251

EFFECTS OF DIETARY SELENIUM SOURCES ON PRODUCTIVE PERFORMANCE, HATCHING TRAITS AND ANTIOXIDANT STATUS OF COMMERCIAL BROILER BREEDERS

S. Ashraf\*(1), S.Bhatti(1), F.Shahid, N.Sahar(1), M.Shoaib(1), H.Iqbal(1)

(1)Institute of Animal and Dairy Sciences, Faculty of Animal Husbandry, University of Agriculture, Faisalabad 38040, Pakistan, Faisalabad, Pakistan

* Corresponding author: DrShahzadBalouch@Yahoo.Com

Objective of the experiment was to evaluate the effects of different dietary sources of selenium on productive performance, hatching traits, antioxidant capacity and immune responses of commercial broiler breeders (Ross-308). A total of 600 female and 180 male, 50-wk-old Ross-308, broiler breeder birds were randomly allotted to 4 treatments, having 5 replicates with 30 females and 9 male birds each, with a 14-d pre-treatment and 98-d trial period. Birds were fed a basal corn-soybean diet supplemented with inorganic i.e. Sodium Selenite (SS; control) and other three groups were fed diet supplemented with. Selenium enriched yeast, L-Seleno-methionine (L-Se-Meth) or Seleno-hydroxy-methionine (OH-Se-Meth), respectively, at 0.3 mg/kg of diet. Egg production and quality parameters, fertility and hatchability were measured as performance indicators. After incubation, chicks of each replicate were individually weighed and graded separately. Glutathione peroxidase activity, total antioxidant capacity and antibodies titer against Newcastle disease virus was also measured. Data collected were analyzed using GLM Procedures of Minitab Statistical Software 18. Means were compared using Tukey's Test. Dietary selenium sources did not affect egg production and their quality parameters, fertility rate, hatchability percentage and antibodies titers against Newcastle disease virus at any phase of experiment. Glutathione peroxidase status, total antioxidant capacity and chick quality were improved by inclusion of organic selenium in the diet of broiler breeders. It was concluded that organic selenium supplementation improved chick quality and antioxidant status, however, did not affect body weight, flock uniformity, egg production and their quality, and immune responses in commercial Ross-308 broiler breeders.

Keys words: Selenium sources, egg production, egg quality, antioxidant status, immune response, broiler breeders
ID: 1261

EFFECT OF WHEAT MALTING ON ZOOTECHNICAL PERFORMANCE OF SLOW GROWING BROILER
CHICKENS

S. Kuenz*(1), K.Damme(2), W.Windisch(1), D.Brugger(3)

(1) Technical University of Munich, TUM School of Life Sciences, Chair of Animal Nutrition, Freising, Germany

(2) Bavarian State Research Center for Agriculture, Department for Education and Poultry Research, Kitzingen, Germany

(3) University of Zurich, Vetsuisse Faculty, Institute of Animal Nutrition, Zurich, Switzerland

* Corresponding author: sylvia.kuenz@wzw.tum.de

In conventional production systems, exogenous phytase is used as feed additive to increase phosphorous (P) digestibility. However, this is not allowed in European organic farming systems. Earlier studies suggest that a germination (malting) is accompanied by significant hydrolysis of phytic acid (Lemmens et al., 2018). The aim of this experiment was therefore to investigate the effect of malted wheat on the zootechnical performance of slowly growing meat chickens under the terms of practical feeding according to the rules of organic farming. Four different diets were formulated for this experiment containing either wheat or malted wheat in combination with and without phosphorus (P) supplementation from 1.5% Monocalcium-phosphate, respectively (diet A: malted wheat + low P; diet B: malted wheat + high P; diet C: wheat + low P; diet D: wheat + high P). 2.000 1-day-old broiler chickens (both sexes) were used for this experiment. From d1 to d16 the birds were fed with an organic starter diet. On d17 the equal number of birds from both sexes were randomly allocated to the four experimental diets (5 replicates per diet, 100 birds per pen) comprising a completely randomized design. Live weight (LW), feed (FC) and water consumption (WC) was recorded pen wise on a weekly basis. Statistical analyzes comprised ANOVA (wheat variant, phosphorus and interactions). From d28 on, birds fed with wheat had a significantly higher LW (760±40g) than birds fed with malted wheat (700±50g; p = 0.0081). Surprisingly, P content had no significant effect on LW and FC during the whole experimental period. But groups fed with wheat had also a significant higher FC (89±5g vs. 81±5g) from d28 onwards (p

References:

Lemmens, E., N. de Brier, K. M. Spiers, C. Ryan, J. Garrevoet, G. Falkenberg, P. Goos, E. Smolders, and J. A. Delcour. 2018. The impact of steeping, germination and hydrothermal processing of wheat (Triticum aestivum L.) grains on phytate hydrolysis and the distribution, speciation and bio-accessibility of iron and zinc elements, Food chemistry. 264:367-376. doi: 10.1016/j.foodchem.2018.04.125.
EFFECT OF A NOVEL 6-PHYTASE AT 250 AND 500 FTU/KG ON PERFORMANCE AND BONE ASH IN BROILERS

L. Nollet\*(1), J.Michiels(2)

* Corresponding author: lode.nollet@huvepharma.com

A broiler trial was set up with 480 male ROSS 308 broilers distributed over 24 pens to investigate the effect of a novel thermostable 6-phytase (OptiPhos® Plus, Huvepharma NV) on performance and tibia bone ash content. All birds were fed a P sufficient starter diet (day 1 to 5) after which different grower feeds (day 5-21) and a finisher feeds (day 22-35) were provided. The starter feed contained 21.5 % CP, 0.90 % Ca and 0.45 % available P (aP). The grower feed contained 20.5 % CP, 0.75 % Ca and 0.34 % aP and the finisher feed contained 19.4 % CP, 0.7 % Ca and 0.31 % aP. A negative control (NC) grower and finisher feed was produced by reducing 0.10 % Ca and 0.15 % aP from the PC. To this NC, 250 and 500 FTU/kg of phytase was added. Feeds were pelleted at ± 80°C (starter was crumbled). Technical performance was measured after every feeding phase. At day 21, 2 birds per pens were selected with a weight close to the pens' average weight and the right tibia was removed, pooled to one sample and analysed for ash on fat free dry matter. The basic statistical models employed were one-way ANOVA, whereby P≤0.05 was considered statistically significant.

Reducing Ca and P from the PC led to reduced end weight (2004 g for the NC vs 2425 for the PC, P<0.05) while FCR was non-significantly higher (1.47 vs 1.44). Adding phytase brought performance up to 2293 g for 250 FTU/kg (sign. different from PC and NC) and 2323 g at 500 FTU/kg (sign different from NC, but not from PC). The FCR were not significantly different from PC and NC (1.46 and 1.45 for 250 and 500 FTU/kg). Tibia ash dropped from 49.4 % (PC) to 41.6 % for the NC (P<0.05), while the addition of the phytase brought bone ash levels up to 45.5 and 46.5 % for 250 and 500 FTU/kg respectively (P<0.05 vs NC). It can be concluded from this trial that OptiPhos® Plus already at low inclusion level of 250 and 500 FTU/kg to a P deficient diet could increase performance of the broilers to the same level as broiler fed a P sufficient diet, while bone ash was also increased.

<footnote>(1) Huvepharma NV, Antwerp, Belgium</footnote>

<footnote>(2) University of Ghent, Ghent, Belgium</footnote>
ID: 133

EFFECT OF A NOVEL E. COLI 6 PHYTASE ON BROILER PERFORMANCE, BONE ASH AND ILEAL DIGESTIBILITY OF PHYTATE, PHOSPHORUS AND PROTEIN AT LEVELS UP TO 1000 FTU/KG

L. Nollet\*(1), K.Kozlowski(2)

A broiler trial was set up (98 floor pens, 17 male ROSS 308 broilers per pen) to investigate the effect of a novel thermostable 6-phytase (OptiPhos® Plus, Huvepharma NV) on performance, bone mineralization and ileal digestibility of total phosphorous (P) and crude protein (CP). All birds were fed a P sufficient starter diet (day 1 to 5) after which different grower feeds (day 5-21) and finisher feeds (day 22-35) were provided. The starter feed contained 21.5% CP, 0.90% Ca and 0.45% available P (aP). The grower feed contained 20.5% CP, 0.80% Ca and 0.32% aP and the finisher feed contained 19.4% CP, 0.75% Ca and 0.30% aP. A negative control (NC) grower and finisher feed were produced by reducing 0.15% and 0.15% Ca and 0.17% and 0.15% aP from grower and finisher, respectively. To this NC, 0, 250, 500, 750 and 1000 FTU/kg of phytase was added. Feeds were pelleted at ± 80°C (starter was crumbled). Feed contained an indigestible marker (0.4% TiO2) in order to calculate digestibility. Technical performance was measured after every feeding phase. At day 21, 2 birds per pens were selected close to the pens' average weight and the right tibia was removed, pooled to one sample and analysed for ash on fat free dry matter. At day 35, 9 birds of each pen were killed to collect ileal material (from terminal ileum to 10 cm before the ileo-cloacal junction) for the determination of P, CP, and Ti. The body weight (BW) and feed conversion (FCR) for the NC were 1.58 kg and 1.57, respectively. Adding phytase increased the BW and FCR in a dose-responsive way reaching 2.30 kg and 1.52, respectively at 35 days (p0.05 vs phytase treatments). Similarly, bone ash was increased from 36.9% to 45.9% at 0 and 1000 FTU/kg phytase (P<0.05 for all phytase levels vs NC), while the PC reached 47.9%. Protein digestibility in the PC and NC were 69.8 and 73.5%, respectively, while protein digestibility increased to 78.9, 79.2, 77.8 and 77.6% for 250, 500, 750 and 1000 FTU/kg phytase, respectively (p<0.05 vs NC and PC at all phytase levels). The addition of phytase to the NC feed increased dig. P by 1.00, 1.12, 1.54 and 1.53 g/kg feed at 250, 500, 750 and 1000 FTU/kg, respectively. It can be concluded from this trial that the addition of OptiPhos® Plus at increasing levels to a P deficient diet increased the performance, bone ash formation and digestibility of P.

<footnote>(1)Huvepharma NV, Antwerp, Belgium</footnote>

<footnote>(2) University of Warmia and Mazury, Olsztyn, Poland</footnote>

<footnote>* Corresponding author: lode.nollet@huvepharma.com</footnote>
TOWARDS PRECISION FEEDING IN LAYING HENS: UPDATE AND VALIDATION OF A MATHEMATICAL MODEL TO PREDICT DAILY CALCIUM AND PHOSPHORUS FLOWS

Y. Guyot\*(1), B. Méda(2), A. Nozeran(2), C. Souchet(1), J. Bernard(2), E. Pampouille(1)

(1) ITAVI, Nouzilly, France

(2) INRAE, Nouzilly, France

* Corresponding author: guyot@itavi.asso.fr

In laying hens, precision feeding could consist in providing an optimal calcium supply (level, source, and distribution kinetics) to ensure the well-being and longevity of animals (i.e. limiting osteoporosis and bone fractures) while maintaining eggshell quality. During eggshell synthesis (occurring mostly by night), hens can mobilize calcium from bones stored in a specific structure called the medullary bone. The calcium stock is then rebuilt after oviposition. However, the animal cannot always fully compensate for a supply deficiency in calcium from feed. An intense and repeated mobilization / reconstitution of the medullary bone can thus lead to long-term and severe lesions affecting the animal's skeleton. To better understand and predict daily calcium requirements, and minimize bone mobilization, an existing mathematical model [1] describing calcium and phosphorus fluxes in the different compartments of the animal (i.e. digestive tract, blood, bone, and egg) was implemented and adapted. The main adaptation of the existing model was to include the ability to simultaneously consider different calcium sources (i.e. meal vs. particles) with different associated solubilization rates. An experimental trial involving 288 Lohmann Tradition laying hens from 19 to 33 weeks of age was conducted. Six different dietary treatments were tested (4 pens of 12 hens/treatment). They differed according to the form of calcium carbonate (i.e. meal, two different particle sizes, or a mix of meal and particles) and to the relative amount of feed distributed in the morning and in the afternoon (30:70 or 70:30). Three sampling series at 27, 30, and 33 weeks of age were performed to measure ionic calcium and inorganic phosphorus blood concentrations over 24 hours (3 to 6 samples per hen). The model was then calibrated and subsequently validated according to these measured concentrations. In its current state, the model can be seen as a decision support system (DSS) for the precision feeding in laying hens, giving to the user an insight on what could be the best feeding strategies to maintain animal longevity, increase well-being or even reduce feed cost, at the scale of an individual hen. In the future, this model could also be implemented in a new tool, taking into account the variability among birds (e.g. oviposition time), to identify the best feeding strategies at the flock level.

[1] Kebreab et al. (2009). Poultry Science, 88 (3), 680–689.
ID: 697

EFFECT OF SUPPLEMENTING CALCIUM PIDOLATE AND PHYTASE IN DIETS WITH LOW LEVELS OF CALCIUM AND PHOSPHORUS ON BROILER PERFORMANCE

B. Iglesias(1), M.Charriere(1), J.Melo*(2)

(1) Sección Avicultura, EEA Pergamino, Instituto Nacional de Tecnología Agropecuaria (INTA), Pergamino, Argentina

(2) UNIVERSIDAD CATOLICA ARGENTINA, Escobar, Argentina

* Corresponding author: jemelo@uca.edu.ar

The calcium-phytate complex may reduce calcium (Ca) absorption and may also reduce the activity of phytase. Decreasing dietary Ca may improve phosphorous (P) utilization, while an excess of Ca may aggravate a P deficiency. Therefore, different researchers have shown that a moderate reduction on dietary Ca had no deleterious effects on broiler performance. Consequently, we tested the hypothesis that a decrease in the level of Ca and P may not affect broiler performance if molecules to improve mineral retention were included in the diet. The aim of this study was to evaluate the effect of Ca pidolate and phytase dietary supplementation on the broilers performance fed low Ca and P diets. A total of 486 day-old male chicks (Cobb-500) were allocated in floor pens in a completely randomized block design (9 replicates/treatment). Pre-starter (1-7 d), starter (8-21 d), grower (22-28 d) and finisher (29-41 d) diets were used for dietary treatments. Control diets (CTRL) were formulated according to Cobb recommendations (2018). Low Ca and P diets (LCP) were formulated to obtain a 20% reduction in total Ca and available P, and no differences in metabolizable energy and the main digestible amino acids. Another set of LCP diets were formulated to include Ca pidolate and phytase (LCP+PP diets). Ca pidolate (PIDOLin PCa, Dietaxion) was included on-top between 0-21 d at 300 ppm. Phytase matrix was used for the formulation of the LCP+PP diets (750 FTU/Kg feed, Axtra PHY, Dupont). Birds and feed were weighed weekly individually and by pen, respectively, and the mortality recorded daily. Data were analyzed as a 2-way ANOVA. The pen was considered the experimental unit. Although there werenot significant differences (P>0.05) in feed intake at the end of the experiment, CTRL broilers had a higher feed intake than LCP and LCP+PP (P0.05). Also for feed conversion ratio there were not significant differences between treatments (P>0.05). Using the performance results at the end of the experiment and the cost of each diet, the feeding cost per kg was calculated and were (USD/kg): 0.271, 0.289 and 0.299 for LCP+PP, LCP and CTRL, respectively (P<0.05). It can be concluded that total Ca and available P concentrations can be reduced by 20% in broiler diets with no effects on their performance. If done in combination with Ca pidolate and phytase allowed to lower the feeding cost.
# EFFECTS OF IN OVO INJECTION OF NANO SELENIUM AND NANO ZINC OXIDE AND HIGH EGGSHELL TEMPERATURE DURING LATE INCUBATION ON HATCHABILITY AND ANTIOXIDANT CAPACITY AND GENE EXPRESSION OF HEAT SHOCK PROTEIN 70 OF BROILER HATCHLINGS

S. Mousavi*(1), M.Shokraneh(2), A.Sadeghi(2), S.Esmaeilkhanian(3), M.Chamani(2)

(1)Azad University, Tehran, Iran

(2) Department of Animal Science, Science and Research Branch, Islamic Azad University, Tehran, Iran, Tehran, Iran

(3) Animal Science Research Institute of Iran, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran, Karaj, Iran

* Corresponding author: snmousavi@hotmail.com

## Introduction

With the onset of pulmonary respiration after internal pipping in the chick embryo, the presence of reactive oxygen species (ROS) and oxidative stress is higher, while natural antioxidant reserves have not reached an adequate level for innate protection (Surai, 2007). Also, it is reported that heat stress cause oxidative stress (Elnesr et al, 2019). Therefore the effect of in ovo injection of Nano-Se and Nano-ZnO and high incubation temperature was evaluated.

## Materials and Methods

On 17 day of incubation (doi), 750 fertile eggs assigned to 5 experimental treatments of 5 replicates including non-injected eggs incubated at normal incubation temperature (37.8°C) or high incubation temperature (38.9°C) and eggs injected with 0.5 ml 0.9% NaCl solution or 40 µg Nano-Se or 500 µg Nano-ZnO in 0.9% NaCl which are incubated at high incubation temperature. Eggs of heat stress group were exposed to high temperature from 19 to 21 doi. Blood samples of 10 hatchlings per treatment were collected and plasma malondialdehyde (MDA) and total antioxidant capacity (TAC) were determined. Total RNA of liver samples were extracted and Real-time PCR was used to evaluate the liver HSP70 gene expression. Data were subjected to the GLM procedure for ANOVA. Means were compared using Duncan's test.

## Results

Percentage hatchability was not affected by treatments. Egg exposure to high incubation temperature reduced TAC and increased MDA of hatchlings plasma. In ovo injection of Nano-ZnO and Nano-Se restored the TAC plasma and reduced MDA concentration. Although embryonic heat stress during late incubation increased gene expressions of HSP70, in ovo injection of the nano minerals ameliorated the adverse effects of heat stress.

## Conclusion

It is concluded that in ovo injection of Nano-ZnO and Nano-Se could reverse negative effects of high incubation temperature during late incubation on oxidative status and liver HSP70 gene expression in broiler hatchlings.

## References

Surai, P.F., 2007. Natural Antioxidants in Poultry Nutrition: new developments. World Poult Sci Ass. Proceedings of the 16th European Symposium on Poultry Nutrition, Strasbourg, France, 26-30 August. Elnesr, S. S., H. A. M. Elwan, Q. Q. Xu, C. Xie, X. Y. Dong, and X. T. Zou. 2019 Effects of in ovo injection of sulfur-containing amino acids on heat shock protein 70, corticosterone hormone, antioxidant indices, and lipid profile of newly hatched broiler chicks exposed to heat stress during incubation. Poultry Science 0:1–9
PHYSIOLOGY OF NUTRITION
COMBINING ALGAL EXTRACTS TO IMPROVE BROILER PERFORMANCE THROUGH INFLAMMATION REDUCTION IN THE GUT

S. Haberecht*(1), J.Roberts(2), S.Wilkinson(3), S.Wu(4), R.Swick(2)

(1)University of New England, Melbourne, Australia

(2)University of New England, Armidale, Australia

(3)Feedworks, Lancefield, Australia

(4)University of New England, Armidale, Australia

The poultry industry is continually under pressure to reduce the use of antibiotic/antimicrobials in feed. Macro algae contain unique sulphated beta-glucan polysaccharides that have shown to be anti-inflammatory and antibacterial when used in animal feed of pigs and poultry (O'Doherty et al., 2010), (Yan et al., 2011). However, their use in poultry is currently limited due to variations in purity, different extraction methods and difficulty in assessing the ratio of the extracts when used together (Fitton, 2011).

The present study was conducted to investigate effects on inflammation markers and performance of broilers. The marine algal extracts, fucoidan (500PPM) and laminarin (300PPM) were fed separately and in combination (500PPM and 300PPM respectively) with a control diet to 1,200 Ross 308 males, half of the birds were then challenged with Salmonella typhimurium at day 8 resulting in 8 treatments. Weight gain analysis for the period 7-14 days, the days immediately post challenge, show that when S. typhimurium challenged birds were fed the fucoidan extract during this time no difference in weight gain to non-challenged birds was observed, where all other treatments had significantly (P=0.045) lower weight gain for the same period compared to the non-challenged treatments. When investigating the inflammation response, adding fucoidan increased calprotectin levels by only 52% for fucoidan alone and 46% for the fucoidan and laminarin combination when compared to the control without challenge, however, the laminarin only treatment significantly (P=0.042) increased calprotectin levels by 72% after the S. typhimurium challenge compared against the unchallenged control. These results suggest a difference in mode of action in response to the S. typhimurium challenge between the two test materials.

These recent works show the potential of high quality marine sulphated polysaccharides, when used in broiler feed, to improve performance through a reduction in inflammation after infectious challenge and via inflammatory responses in the gut.

References:

Fitton, J. H. 2011. Therapies from Fucoidan; Multifunctional Marine Polymers. Marine Drugs 9(10):1731.

O'Doherty, J. V., P. McDonnell, and S. Figat. 2010. The effect of dietary laminarin and fucoidan in the diet of the weanling piglet on performance and selected faecal microbial populations. Livestock Science 134(1):208-210. doi: https://doi.org/10.1016/j.livsci.2010.06.143

Yan, G. L., Y. M. Guo, J. M. Yuan, D. Liu, and B. K. Zhang. 2011. Sodium alginate oligosaccharides from brown algae inhibit Salmonella Enteritidis colonization in broiler chickens. Poultry Science 90(7):1441-1448. doi: 10.3382/ps.2011-01364

<footnote>* Corresponding author: shaberec@myune.edu.au</footnote>
GROWTH HORMONE, INTESTINAL MORPHOLOGY AND JUVENILE GROWTH OF BROILER CHICKENS FED DIFFERENT LEVELS OF CURCUMA LONGA UNDER HOT HUMID TROPICAL CLIMATE

O. Oke*(1), T.Lawal(2), J.Abiona(3), S.Rahman(3)

(1)Regional Centre for Poultry Science (CERSA), University of Lome, Togo, Lome, Togo

(2)Federal University of Agriculture, Abeokuta, Nigeria, Abeokuta, Nigeria

(3)Federal University of Agriculture, Abeokuta, Abeokuta, Nigeria

* Corresponding author: emaoke7@yahoo.co.uk

The performance of broiler chickens in the tropical environments in the absence of temperature-controlled housing facilities is suboptimal as a result of thermal stress. Nutritional strategy has been proposed as a viable means of mitigating the adverse effect of thermal stress on the birds to reduce economic losses. This study, therefore aimed at evaluating the juvenile growth, growth hormone and intestinal morphology of broiler chickens fed different levels of curcuma longa under hot humid tropical climate. Two hundred and forty broiler chicks were randomly allocated to four dietary treatments of basal diets supplemented with 0 (CT), 4 (TG), 8 (FT) and 12g (SG) of turmeric powder/Kg of diet, having 4 replicates of fifteen birds each, in a Completely Randomized Design for 8 weeks. Data were collected weekly on feed intake and body weights at the juvenile age. Blood samples were collected from 8 birds per treatment at week 3 (starter phase) and week 8 (finisher phase) of the experiment for the determination of plasma growth hormone. At day 56 of age, 8 birds per treatment were slaughtered and dissected. 2cm segments of duodenum, jejunum and ileum were collected for the measurement of villi width, villi height and crypt depth and villi height: crypt depth ratio. Data obtained were analyzed using one-way ANOVA using SAS (2008). Results showed that the weight gain of the chicks in SG and TG were however comparable to that of CT but the weight gain of the chicks in FT was higher than that of CT. The feed intake of the birds was similar across the treatment group at week 2. FCR of the birds in CT was similar to those of TG and SG but higher than that of FT. Plasma growth hormone was similar at starter and finisher phases. Duodenal villi width of the birds in SG, TG and CT was similar but lower than that of FT. The ileal crypt depth of chickens in FT was lower than those of CT but similar to those of the other treatment groups. There was no difference in the crypt depth of jejunum. The ratio of villi to crypt depth ratio in the duodenum was in the order of FT > SG > TG > CT. The ratio in ileum was higher in FT than that of TG which was higher than SG whose value was higher than that of CT. It was concluded 8g/Kg diet Curcuma longa improved juvenile growth and nutrient absorption through enhanced intestinal morphology of broiler chickens under hot humid tropical climate.
THE EFFECT OF FUMONISIN CONTAMINATION ON BROILERS IMMUNE RESPONSE AFTER VACCINATION

A. Koppenol\*(1), M.Ingberman(2)

(1)Impextraco NV, Heist op den berg, Belgium

(2)Imunova, Curitiba, Brazil

* Corresponding author: astrid@impextraco.be

There is a market request for feed additives that help to overcome immunosuppression by mycotoxins after vaccination. To determine immunosuppression by fumonisins (FUM) after vaccination and to screen for a solution, a 3x2 full factorial design was setup. In total, 330 Ross-308 birds (5T*6R*11) were randomly allocated to: (1) a control diet; (2) a contaminated diet with 20 ppm FUM; (3) a contaminated diet with 100 ppm FUM; (4) a contaminated diet with 20 ppm FUM supplemented with Elitox® at 0.25% or (5) a contaminated diet with 100 ppm FUM supplemented with Elitox® at 0.25%. At d7, 2 birds per pen were subcutaneously vaccinated against Newcastle disease with a killed vaccine (Poulvac, Zoetis). Blood samples were taken from 2 vaccinated and 2 non vaccinated birds per pen at d28 to determine the humoral immune response (major histocompatibility complex II (MHCII) expression, Immunoglobulins and Antigen Presenting Cells (APC)), and cell-mediated immune response by interferon gamma (IFN-y). Vaccination resulted in higher percentages of APC (p=0.0262) and monocytes (p<0.0001), but lower levels of MHC II expression by APC (p=0.0026). Whereas, increasing levels of FUM resulted in decreasing levels of APC (p=0.0456) and monocytes (p=0.2012). This clearly indicates the adverse effects of mycotoxins on the humoral immune response after vaccination. Low levels of FUM (20 ppm) resulted in decreased levels of MHC II expression, whereas contamination with higher levels (100 ppm FUM) resulted in increased MHC II expression by monocytes (p=0.0041) and APC (p<0.0001). This suggests that more energy is allocated towards immunity in the highly contaminated birds, as confirmed by the numerically worse performance. The amount of IFN-y was also significantly increased in blood of birds receiving 100 ppm FUM (p=0.0001), indicating that FUM at such high levels promotes inflammation with its repercussion on performance. Supplementation of Elitox® not only resulted in numerically heaviest animals but also restored contaminated birds humoral and cell-mediated immune response after vaccination, by reducing the increased IFN-y at 100 ppm (p=0.0001) and by increasing MHC II expression at 20 ppm FUM (p=0.0089). In general it can be concluded that FUM had an inverse effect on vaccination and that Elitox® could prevent the losses induced by FUM contamination.
DIETARY SUPPLEMENTATION WITH A MICROENCAPSULATED BLEND OF ORGANIC ACIDS AND BOTANICALS ALTERS METABOLIC PATHWAYS IN THE JEJUNUM TO ENHANCE GROWTH AND FEED EFFICIENCY IN BROILERS

C. Swaggerty*(1), R.Arsenault(2), A.Piva(3), E.Grilli(3)

(1)U.S. Department of Agriculture, Agricultural Research Service, College station, United States

(2)University of Delaware, Newark, United States

(3)University of Bologna and Vetagro S.p.A., Bologna and regio emilia, Italy

* Corresponding author: christi.swaggerty@usda.gov

A microencapsulated feed additive containing citric and sorbic acids, thymol, and vanillin (AviPlus®P) has been shown to increase growth rate and feed efficiency in broilers; however, the mechanisms associated with these benefits are not fully understood. The objective of this study was to identify key metabolic signaling pathways activated by dietary supplementation with this blend of organic acids and natural botanicals. Day-of-hatch by-product breeder chicks were randomly assigned to a control diet or a diet supplemented with 500 g/metric ton AviPlus®P (n=2 replicate pens/group). At 15-d-of-age, a section of jejunum was collected for kinome analysis using an immuno-metabolism peptide array to compare control and treated tissues (n=5 birds/pen [10 birds/treatment were used for the analyses]). The data was analyzed using PIKA2 peptide array analysis software and fold-change between control and treated groups determined. The P value was calculated by conducting a one-sided paired t-test between treatment and control values for a given peptide. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was performed using Search Tool for the Retrieval of Interacting Genes (STRING; https://string-db.org). Changes to key metabolic pathways include: Insulin, PI3k-Akt, AMPK, FoxO, mTor, and adipocytokine signaling pathways. Additional analysis of the AMPK and insulin signaling pathways revealed numerous peptides with increased (P<0.05) levels of phosphorylation. Specific proteins that were associated with these peptide changes in the AMPK (5' adenosine monophosphate-activated protein kinase) pathway included PFK and G6Pase which impact glycolysis; ACC and CPT1 which are involved in fatty acid oxidation; or CyclinD which can result in growth arrest. The Insulin signaling pathway had increased phosphorylation of IRS, PDK1/2, TSC2 and 4EBP1 which can lead to protein synthesis and reduced cell growth. The changes in these signaling pathways provide insight into how supplementation specifically impacts energy use and production in a segment of the gut. Collectively, the kinome analysis showed key differences in the jejunum sample from the supplement-fed birds compared to those on the control diet. The data provide insight into the mechanisms and pathways associated with increased growth rate and feed efficiency in broilers following supplementation with a microencapsulated feed additive containing organic acids and natural botanicals.
BIOCHEMICAL BLOOD PARAMETERS IN LAYING HENS FED DIETS SUPPLEMENTED WITH DIFFERENT VEGETABLE OILS

A. Grozina*(1), V.Vertiprakhov(1), V.Fisinin(1)

(1)All-Russian Research and Technological Poultry Institute of Russian Academy of Sciences, Sergiev posad, Russia

* Corresponding author: alena_fisinina@mail.ru

The experiment was performed on 10 Hisex White hens (28-46 weeks of age) fed a diet optimized according to the recommendations for this. The birds were allotted to four treatments: control treatment 1 was fed the basic diet supplemented with 2.6% of sunflower oil; the latter was substituted in the experimental treatments 2-4 by soybean, rapeseed, and flaxseed oils, respectively. The blood was sampled (2-3 ml) from the axillary vein after 14 hours of starving, the solution of sodium citrate was added. The samples were centrifuged at 5000 rpm for 5 min. The activities of the digestive enzymes in serum were determined on semiautomatic biochemical analyzer BS-3000P (Sinnowa, China) and automatic analyzer ChemWell 2900 (T) (USA) using reagent kits Human (Germany). The tryptic activity was determined with benzoyl-DL-arginine p-nitroanilide (BAPNA) as a substrate. All experiments were performed in 5 replicates.

The tryptic activity in serum in treatments 2 and 3 was significantly higher in compare to control by 84.3 and 87.6%, respectively (P

The study was financed by the sub-program "The investigation of adaptive mechanisms in the digestive system in mammals and poultry to the diets with different ingredients" implemented by the Russian Academy of Sciences (The Presidium's Resolution No 132, July 5, 2017).
# THE ADAPTATION OF EXOCRINE PANCREATIC FUNCTION IN LAYING HENS TO DIETARY LIPID INGREDIENTS

V. Vertiprakhov\*(1), A. Grozina(1), V. Fisinin(2)

(1) All-Russian Research and Technological Poultry Institute of Russian Academy of Sciences, Sergiev posad, Russia

(2) All-Russian Research and Technological Poultry Institute of Russian Academy of Sciences, Sergiev posad, Russia

\* Corresponding author: alena_fisinina@mail.ru

The use of different vegetable oils in poultry nutrition is still an urgent problem. The published data of the research on broiler chicks evidence that different oils can affect the metabolism and growth efficiency. However, the mechanism of the influence of different vegetable oils on the exocrine pancreatic function is still understudied. The physiological study presented was performed on three Hisex White laying hens (Gallus gallus L.) with chronic fistulae of main pancreatic duct inserted using the method of Batoev (1970). The method of periods (7-10 days) was used. In the control period the hens were fed standard layer diet supplemented with sunflower oil; in experimental period 1 the sunflower oil was substituted by soybean oil; in period 2 – by rapeseed oil; in period 3 – by flaxseed oil. The amount of pancreatic juice released during the collection period and enzymatic activities within the juice were determined.

It was found that lipase activity is adapting to the oil used: the highest lipase activity was found in control (with unrefined sunflower oil); the increase in lipase activity was accompanied by the increase in the amylase activity. The postprandial dynamics of activity of the pancreatic lipase evidenced that sunflower and flaxseed oils featured better palatability for the hens: during the first 60 min after the feeding (the period corresponding to complex-reflex phase of regulation of the exocrine pancreatic secretion) the enzymatic activity grew 1.9-2.5-fold with these two oils while two other oils (soybean and rapeseed) produced the relatively substantial increases in lipase activity during the neurohumoral phase of the regulation evidencing the better nutritive value of these oils. The correlation between the activities of proteases and alkaline phosphatase in the pancreatic juice was found; a phosphatase-protease index can be therefore proposed for the assessment of response of the digestion to alterations in diet composition. The conclusion was made that the changes in the exocrine pancreatic function in hens fed diets with different vegetable oils can be the basis for further research of the digestion in poultry to identify optimal dietary ingredients.

The study was financed by the sub-program "The investigation of adaptive mechanisms in the digestive system in mammals and poultry to the diets with different ingredients" implemented by the Russian Academy of Sciences (The Presidium's Resolution No 132, July 5, 2017).
BACILLI CAN CREATE A PROTECTIVE BIOFILM ON THE EPITHELIUM

C. Bostvironnois(1), D.Sandvang*(2), J.Schleifer(3)

(1)Chr Hansen, Arpajon, France

(2)Chr Hansen, Horsholm, Denmark

(3)Chr Hansen, Milwaukee, United States

* Corresponding author: frcbos@chr-hansen.com

Bacillus-based probiotic products containing bacterial spores seem to be particularly well-suited for use in broiler feeds, as they are metabolically dormant and resilient to environmental stresses, including pelleting. There have been years of debate on the mode of action of probiotics in chickens and more broadly in poultry. One of the cornerstones of the debate was the ability of spores to germinate and become viable organisms in the intestine due to the rapid transit time in poultry. These aspects were clarified in 2008 by Cartman et al. The research has shown that orally-administered Bacillus subtilis spores germinate in the chicken's gastro-intestinal tract (GIT) (Cartman et al. 2008). Continuous administration of an effective Bacillus subtilis probiotic is advisable to achieve consistent benefits (Latorre et al. 2014). However, another point of discussion was to know whether spore-forming Bacilli were transient organisms in the gut or if they could attach somehow to the intestinal epithelium. This led to two schools of opinion in the scientific community.

The study was conducted to investigate the effects of commercially-available spores of Chr. Hansen Bacillus subtilis spore-based probiotic in diets at 1.6 × 10⁶ cfu/gram of feed on performance and microbiota activity in broilers. Fluorescence in situ hybridization (FISH) was performed to investigate the spatial organization and the formation of Bacillus subtilis biofilms in intestinal samples from various GIT locations in 6 broiler chickens. Tissue sections from each chicken were analyzed in duplicate and visualized by fluorescence microscopy with a 40x objective, using a fluorescence microscope.

Indeed, a first picture describes very well how Bacillus colonize the intestinal epithelium in the intestine. It can be seen very clearly the red fluorescence on the surface of the villi of the intestine. In a second picture with a different fluorescence, we can also see some luminescence inside the lumen of the intestine, which clearly shows that transient Bacillus are in the intestine, live and multiplying into the lumen of the gut content. This most recent research helps to further understand an important part of the mode of action of probiotics, such as Bacilli. Bacilli can germinate in the gut, being an active part of the bacteria microbiome in poultry, they can be transient, live organisms in the flow of the intestinal content, and can colonize the surface of the intestinal villi.
EFFECT OF GANODERMA LUCIDUM EXTRACT ON THE FECAL MICROBIOTA AND BURSAL TRANSCRIPTOME OF BROILERS

Y. Yu*(1), Y.Cheng(1), W.Chen(1), K.Hua(1)

(1)National Ilan University, Yilan city, Taiwan

* Corresponding author: yuyh@niu.edu.tw

This study investigated the effects of Ganoderma lucidum extract on the fecal microbiome and bursal transcriptome of broilers. A total of 60 one-day-old male broiler chicks (Ross 308) were randomly assigned into 2 dietary treatments, with 5 replicate cages per treatment and 6 birds per cage. The dietary treatments consisted of a basal diet as control and a diet of control plus 1 g/L G. lucidum extract in drinking water. The average body weight, average daily gain, average daily feed intake, and feed conversion ratio were calculated from days 1 to 35. On day 35, feces and bursa of Fabricius from broilers were freshly collected and used for fecal microbiota and transcriptome analysis, respectively. Student's t test was used for intergroup comparison, and a P value less than 0.05 was considered significant. No significant difference was observed in growth performance in broilers after feeding with G. lucidum extract. However, the microbial richness and evenness was reduced in the fecal content of G. lucidum extract-treated group. Principal coordinate analysis demonstrated good clustering of the pair of samples collected from the fecal content of the control group and G. lucidum extract-treated group. The abundance of genus Lactobacillus in feces increased in broilers fed G. lucidum extract, whereas the abundance of genus Ruminococcus torques group in feces was decreased in response to G. lucidum extract treatment. Transcriptomic analysis demonstrated that 2085 genes were differentially expressed in the bursa of Fabricius and that 1114 genes were upregulated and 974 genes downregulated in response to G. lucidum extract stimulation. Gene ontology biological processes related to immune system process, regulation of immune system process, cell activation, and organelle assembly were downregulated in response to G. lucidum extract treatment. A comparative functional analysis results demonstrate that G. lucidum extracts positively regulated T cell receptor signaling pathway, glycerophospholipid metabolism, and N-glycan biosynthesis in the bursa of Fabricius. These results demonstrate that G. lucidum extract can regulate gut microbiota and immune response in the bursa of Fabricius in broilers.
# THE DUODENAL ACTIVITIES OF THE DIGESTIVE ENZYMES IN CHICKEN FED DIFFERENT LIPID DIETARY INGREDIENTS

A. Grozina* (1), VölvVertiprakhov (1), V.Fisinin(1), I.Kislova(1)

(1) All-Russian Research and Technological Poultry Institute of Russian Academy of Sciences, Sergiev posad, Russia

* Corresponding author: alena_fisinina@mail.ru

The most intensive hydrolysis of dietary nutrients occurs in the duodenum. Since the digestion can adjust to the diet composition different vegetable oils with different fatty acid profiles can produce different responses. The aim of our study was the investigation of duodenal activities of the digestive enzymes in chicken fed different oils. Three Hisex White laying hens with chronic duodenal fistulae and method of periods (7-10 days each) were used. The hens were fed a standard diet for layers. In control period the diet was supplemented with sunflower oil (SFO); in subsequent periods the latter was substituted by soybean (SBO), rapeseed (RSO), and flaxseed oils (FSO). The samples of duodenal digesta were taken in 1 postprandial hour. Activity of amylase was determined using Smith-Roy method modified for high activity; activity of proteases by hydrolysis of Hammersten's casein with colorimetric control (450 nm); activities of lipase and alkaline phosphatase (AF) and concentration of calcium were determined on semi-automatic biochemical analyzer BS-3000P (Sinnowa, China) with a reagent kit by DIAKON-VET (Russia).

It was found that the activities of lipase and AF and Ca concentration are the responsive to the shifts in dietary oil ingredients. The highest lipase activity was found in the period with SFO. Substitution of the latter by SBO significantly decreased activity of lipase by 35.6%, for RSO by 37.6%, for FSO by 35.9% (P<0.05). The highest Ca concentration was found with FSO, higher by 12.5% (P<0.05) in compare to SFO; the increases in Ca concentration with SBO and RSO were 10.4 and 12.5% (P<0.05). The AF activity also significantly increased with these substitutions: by 92.6% for SBO, by 29.0% for RSO, and by 46.0% for FSO (P<0.05). The most conclusive parameter characterizing the adjustment of the digestion to diet composition is a phosphatase-protease index calculated as the ratio of the activities of AF and proteases (u/L). Optimal values of this index were found for SFO and RSO; these data are in accordance with our previous research on the pancreatic secretion rates for these oils. The indices for SBO and FSO were 2.1-fold and 1.8-fold higher in compare to the control SFO period.

The study was financed by the sub-program "The investigation of adaptive mechanisms in the digestive system in mammals and poultry to the diets with different ingredients" implemented by the Russian Academy of Sciences (The Presidium's Resolution No 132, July 5, 2017).
INFLUENCE OF DIFFERENT FIBER SOURCES ON THE ILEAL MORPHOLOGY IN BROILER CHICKENS

M. Liebl\*(1), M. Gierus(2), C. Potthast(3), E. Schneeberger(4), K. Schedle(2)

(1) FFoQSI GmbH, Vienna, Austria

(2) University of Natural Resources and Life Sciences (BOKU), Vienna, Austria

(3) Agromed Austria GmbH, Kremsmünster, Austria

(4) Garant-Tiernahrung GmbH, Pöchlarn, Austria

\* Corresponding author: mariella.liebl@boku.ac.at

The beneficial effects of fibre are well described in the feeding of monogastric animals. Recently the use of fibre in the diet of broiler chicken is a topical issue. Depending on fibre properties, the inclusion of moderate levels of fibrous material can positively influence the development and functionality of the gastrointestinal tract. In this respect, also its epithelial mucosa can be affected. This may increase the digestibility of nutrients leading to an enhanced performance of the chicken (Jimenez et al. 2013). The current study examines the effect of various dietary fibre sources on morphometric parameters of the ileum in broiler chickens.

A total of 6,300 day-old broiler chicken (Ross 308) were reared to 36 days of age on a 3 phase feeding program. The animals were randomly allotted to one of four iso-caloric and iso-nitrogenous treatments: Treatment 1 was a commercial diet for broilers consisting of corn, wheat and soybean meal without a supplemented fibre source, treatment 2 included lignocellulose I, treatment 3 included lignocellulose II and treatment 4 soybean hulls.

After removing the ileal segment, tissue samples from 2 cm to the ileocecal junction were taken and prepared for paraffin embedding. After sectioning and staining with alcian blue and periodic acid-Schiff six villi, crypts and muscular layers of each sampled animal (n=72) were examined with computerized light microscopy.

Statistical analysis of the gathered data was conducted using Statistical Analysis Software (SAS ®, 9.4.). The results show that adding fibre to the diet significantly influences the morphometry of the ileum. The control treatment without any fibre supplementation showed significant shorter villi, lower crypt depth as well as a tendency towards thinner mucosal layer. An increased villus and crypt surface area is important for an improved nutrient absorption surface. Furthermore, the least amounts of goblet cells were detected in the control treatment. An increased amount of mucus building goblet cells can be evaluated positively, as the mucus layer acts cytoprotective and influences the nutrient transport mechanism. This research shows that the fibre supplementation to the diets of broiler chicken positively affects the intestinal morphology in the ileal section.

Jimenez, E.; Frikha, M.; Coca, A.; García, J.; Mateos, G (2013): Oat hulls and sugar beet pulp in diets for broilers-Effects on growth performance and nutrient digestibility. Anim Feed Sci Tech, 182:33-43.
ID: 319

# IS SELECTION FOR IMPROVED PERFORMANCE LIMITING THE GASTROINTESTINAL CAPACITY OF MODERN BROILERS?

J. Taylor\*(1), P. Sakkas(2), I. Kyriazakis(1)

(1) Newcastle University, Newcastle, United Kingdom

(2) Newcastle University, Rennes, France

\* Corresponding author: j.taylor@ncl.ac.uk

Genetic selection for improved carcass yield may limit the capacity of the gastrointestinal tract (GIT) of modern broilers to deal with bulky feeds. Interest has grown around the use of alternative feed ingredients in broiler diets, which are often bulky materials. Reductions in the capacity of the GIT will lead to a limitation in feed intake and consequently performance on such diets. We investigated the capacity of male Ross 308 broilers to deal with increasing levels of dietary bulk and aimed to identify a feed bulk dimension related to limited feed intake (FI). Two bulky ingredients were selected: oat hulls (O), and sugar beet pulp (S), due to the differences in their physicochemical properties. 480 day-old broilers were allocated in 48 pens and offered a common starter until d8 and 1 of 7 feeds from d8-d36 of age; a control feed (C), which was diluted to three levels (15%, 30% or 45%) with either O or S. FI was measured daily and two birds per pen were dissected for organ measurements and empty carcass weight (ECW) at d36. Cumulative FI (CFI) was calculated over the experiment and scaled relative to ECW at d36. It was hypothesised that bulky feeds would increase the visceral weight of the birds, therefore ECW was used as a scalar for CFI (g/ kg CW). Results were analysed with GLM using the nlme package in R.

Broilers were able to accommodate a greater increase in CFI on the O rather than S feeds ($P < 0.001$). CFI (g) of the birds offered C, O15, O30, O45, S15, S30 and S45 were 2683, 3678, 3977, 3795, 3521, 3041, 2808 (±178) g. ECW was reduced by the inclusion of S, whereas the inclusion of O was limiting only at the 45% level ($P < 0.001$): 2486, 2610, 2498, 2211, 2262, 1876, 1301 (±35.1) for birds offered C, O15, O30, O45, S15, S30, and S45 feeds respectively. Scaled CFI were 1084, 1444, 1593, 1814, 1702, 1816, 1956 (±64) g/ kg ECW, respectively. Scaled CFI was increased to a greater extent in the S and O45 birds than the O15, O30 and C birds ($P < 0.001$). The reductions in performance of the S and O45 birds were consistent with the limitations in FI, as they are unable to increase FI to the extent that they meet the nutrient requirements for optimum growth. Nonetheless, our data show that modern broilers can substantially increase FI in response to bulky feeds, almost double their FI on feeds high in S. However, we were unable to identify the property of bulky feeds which was responsible for limiting FI and consequently performance.
IMPROVEMENT OF BROILER PERFORMANCES BY PROBIOTIC IN DIET IS LINKED TO EARLY LIFE
INTESTINAL TRACT DYNAMIC CHANGES

P.Belloir*(1), J. Kamphuls (2), JJ. Plauzolles(3), G.Wallois(3), B. Florence(4), E. Chevaux (4), H.Eutamène(2)

(1) E.I Purpan, Toulouse cedex 3 - france, France

(2) UMR 1331, Toxalim, INRA/INP/UPS, Neurogastroenterology and nutrition team, Toulouse, France

(3) E.I Purpan, Domaine expérimentale de Lamothe, Seysses, France

(4) E.I Purpan, Domaine expérimental de Lamothe, Seysses, France

Probiotics are currently proposed as an alternative solution to antibiotics used in poultry farming. Probiotic strains are described to improve zootechnical performances and are known as healthy beneficial microorganisms by correcting gut microbiota imbalance and gut barrier defect through mucus production stimulation. This study was aimed to determine the incidence of Bacillus pumilus in diet during the rearing period of broiler i) on the zootechnical performances and ii) according to the age, on intestinal tract dynamic changes.

204 broilers Ross PM3 (8 or 9 animals/cage) were used from day (d)0 to d35 and received diet enriched in Bacillus pumilus (Bp group: 1x10^9 CFU/kg feed) or the respective diet alone (control group) (12 cages/treatment). To determine the average daily gain and the feed intake, animals and feed were weighted for 4 periods: d0 to d10 (starter), d10 to d25 (grower), d25 to d35 (finisher), and d0 to d35 (total period). Histological analysis (muscular and mucus layer thickness and goblet cells number) were performed on the ileum at d10 and d35. For statistical analysis, a mixed model was used, treatment was considered as a fixed effect and cage as a random effect.

Compared to control group, no impact of Bp was measured on body weight and average daily gain. From d10 to d25, daily feed intake tended to decrease (p=0.07) with Bp. Feed conversion ratio was significantly reduced by Bp for the 2 periods d10-d25 (p=0.039) and d0-d35 (p=0.015). At d10, in ileum of Bp animals, a thicker gut muscle layer (p=0.07) as well as increased intraluminal mucus production (p=0.004) were measured, compared to control group. At d35 the mucus production in ileum was decreased in Bp group compared to control group (p = 0.004). At d10 and d35, no incidence of Bp was noticed on goblet cells number.

Bacillus pumilus supplemented diet contributes to improving broilers zootechnical performances: FCR improvement was mainly driven by a decrease of feed intake. This beneficial economic aspect observed with Bp diet is associated with a clear impact of Bp during early life period. These data are associated to a more important thickness of intestinal muscular layer in favor of better contact between diet and digestive enzymes and to a reinforcement of the gut barrier illustrated by the higher production of mucus by goblet cells. Finally, Bacillus pumilus seems to be a good candidate to counteract intestinal disorders like malabsorption syndrome in chicken.

<footnote>* Corresponding author: pauline.belloir@purpan.fr</footnote>
INVESTIGATING THE COMBINED EFFECT OF MEDIUM CHAIN FATTY ACIDS AND PHYTOGENIC MOLECULES ON BROILER DIGESTIBILITY

M. Dewez\*(1), J. Vervloesem\*(2), Q. Duan(2), D. Hermans(2), S. Leleu(3), E. Delezie(3), E. N'guetta(4), H. Carmans(2)

(1) Nuscience N.V., Drongen, Belgium

(2) Nutrition Sciences, Drongen, Belgium

(3) ILVO, Melle, Belgium

(4) Agrifirm, Drongen, Belgium

\* Corresponding author: marine.dewez@nusciencegroup.com

Investigating the combined effect of medium chain fatty acids and phytogenic molecules on broiler digestibility

Marine Dewez1, Jan Vervloesem1, Qingyao Duan1, Eric N'guetta1, Hannes Carmans1, Saskia Leleu2, Evelyne Delezie2, David Hermans1

1) Nutrition Sciences NV, 9031 Drongen, Belgium

2) ILVO, Animal Sciences Unit, 9090 Melle, Belgium

Dietary medium chain fatty acids (MCFAs) have strong antibacterial activities and a positive impact on intestinal morphology in broilers, while phytogenic components have antioxidant, anti-inflammatory and enzyme stimulating properties. Improving gut health results in optimizing the nutrient absorption capacity and the efficiency of the bird. Here, we evaluated the effect of a MCFAs and phytogenic mixture (M-prove®, Agrimprove) on broilers' digestibility and its in vitro antibacterial effect. A digestibility study was performed at ILVO. 108 male day-old broiler chicks (Ross 308) were set in deep litter pens and fed a standard wheat-soy based mash diet ad libitum from day 1 to 9. On day 9, the number was reduced to 96 birds, randomly transferred to digestibility units and fed nearly ad libitum (12 replicates per diet, 4 birds per unit): one control diet and one topped with 1.2 kg/ton of the mixture.

Between day 14 and 18, total feed intake and total excreta were determined. Body weight gain was recorded during the collection period. Digestibility was calculated on the basis of the total excreta/feed ratio. Crude protein (CP), crude fat (CF), gross energy (GE) and dry matter digestibility as well as energy contents (metabolizable energy corrected for nitrogen retention; MEn) were determined. Digestibility parameters were analysed with a linear mixed model with diet treatment as a fixed factor, followed by a Tukey post-hoc comparison of means. The mixture tended to slightly increase broilers' feed intake (P=0.064) and significantly increased daily weight gain by 2.5g (P0.1). An in vitro model mimicking poultry gizzard was used (pH=3, 40 min incubation time, co-culture of three bacteria and 3 replicates) and the mixture was added at 0.4 kg/ml medium (= equivalent in vivo dose taking into account a water/feed intake ratio of 2/1). The data in the model showed a decrease of the relative ratio of pathogenic to beneficial bacteria as enumerated via plate counting. Indeed, Enterococcus displayed over 1 log reduction, while Lactobacillus remained unchallenged and an intermediate effect was seen on E. coli. It is concluded that the impact of the combination of MCFAs and phytogenic molecules on digestibility and microbiota profile should be further investigated.

Key words: broiler, fatty acids, phytogenic, digestibility, antibacterial
HYDROXY-METHIONINE OLIGOMERS ARE EFFICIENTLY USED BY BROILERS TO SUSTAIN GROWTH PERFORMANCE.

Y. Mercier\*(1), D.Batono-alavo(2), A.Petrelli(3)

(1)Adisseo France S.A.S, Commentry, France

(2)Adisseo France SAS, Montreal, Canada

(3)Adisseo France S.A.S., Lyon, France

\* Corresponding author: yves.mercier@adisseo.com

Synthetic methionine (Met) sources are currently added in broilers' feed to fulfill sulfur amino acids (SAA) requirements. Despite the wide use of both DL-Methionine (DL-Met) and Hydroxy-Methionine (HMTBA), there is still a debate on their relative bio-efficacy. Liquid HMTBA product presents a natural equilibrium between monomeric and oligomeric forms. The latter are claimed to be less absorbed by birds that would partly explain the difference of efficacy between DL-Met and HMTBA. The aim of this study was to compare birds' growth performance when fed with increments of HMTBA oligomers from 23 to 84%. Birds were fed from 0 to 8 days a commercial starter diet. From 8 to 21 days, 112 Ross 308 male birds were randomly allocated to 6 treatments consisting of a negative control diet, without Met supplementation (dSAA/Lys = 0.54), a positive control supplemented with DL-Met to reach SAA requirement (dSAA/Lys = 0.78) and 4 treatments supplemented on iso-molecular basis with: Liquid HMTBA (23% oligomers), and oligomers enriched products containing respectively 50, 75 and 84% oligomers to reach the same SAA as the positive control diet. The oligomers enriched products were obtained by vacuum water elimination to shift the chemical equilibrium to oligomeric forms. The total HMTBA and oligomeric contents were measured by HPLC. Birds were reared in cages of 2 birds. Animals and feed were weighted at 14 and 21 days. Data were statistically analyzed using a two-way ANOVA with the treatment and the block as main variables. Treatment means were compared using the Fisher test. Globally, feed intake was lower in the negative control (P<0.05) and similar between other treatments. Body Weight Gain and Feed Conversion Ratio were significantly impaired (P0.10) ofbody weight, weight gain and feed conversion ratio were observed between the positive control and other treatments. DL-Met containing no oligomers was similar in performance to all treatments containing up to 84% oligomers (P>0.10). A linear regression applied to only HMTBA treatments also indicated no significant effect of the oligomers level. These results highlight that modern broilers can hydrolyze, absorb and utilize HMTBA oligomers even at high level. According to these results, oligomers level in commercial HMTBA cannot be considered as limiting and reinforce the 100% relative efficacy of Hydroxy-methionine.
ID: 469

CO-EXPOSURE TO DEOXYNIVALENOL (DON) AND CAMPYLOBACTER JEJUNI INCREASES INTESTINAL PERMEABILITY AND BACTERIAL TRANSLOCATION IN BROILER CHICKENS

D. Ruhnau*(1), C.Hess(1), B.Doupovec(2), D.Schatzmayr(2), M.Hess(1), W.Awad(1)

(1)Clinic for Poultry and Fish Medicine, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine, Vienna, Austria, Vienna, Austria

(2)BIOMIN Research Center, Technopark 1, 3430 Tulln, Austria, Tulln, Austria

* Corresponding author: daniel.ruhnau@vetmeduni.ac.at

The mycotoxin deoxynivalenol (DON) is a great health concern in poultry production. DON affects the epithelial cells of the gastrointestinal tract and contributes to the loss of the epithelial barrier function. Similarly, it is debated that C. jejuni negatively affects the integrity of the intestinal epithelium and promotes the translocation of bacteria from the gut to inner organs. So far, no data are available in regard to the co-exposure to DON and C. jejuni in broilers, two widespread foodborne zoonotic agents. Therefore, the aim of the present study was to investigate whether DON interacts with C. jejuni colonization/translocation.

To this end, a total of 120 commercial broiler chickens (Ross 308) were housed in floor pens on wood shavings with feed and water being provided ad libitum. Birds were allocated to four groups (n = 30 with 5 replicates/group) and fed for 5 weeks with either a contaminated diet (5 mg DON/kg feed) or a basal diet (control). Following oral infection of two groups at 14-days of age with 1 x 10⁸ CFU of C. jejuni NCTC 12744, birds were euthanized and tissue samples were collected in weekly intervals until 3 weeks post infection. We investigated the effect of DON and C. jejuni on the epithelial paracellular permeability of duodenum, jejunum and cecum by measuring the mucosal to serosal flux of 14C-mannitol (a paracellular marker) in an Ussing Chamber. Furthermore, the translocation of C. jejuni and E. coli was investigated by determining CFU counts from duodenum, jejunum, cecum, liver and spleen. It was found that the co-exposure of broilers to DON and C. jejuni increased the paracellular permeability of duodenum, jejunum and cecum compared to the individual treatments. In agreement with this, this co-exposure also promoted the translocation of C. jejuni and E. coli to inner organs with a significantly higher bacterial load in liver and spleen. Finally, a reduced growth performance of birds was detected underlining the negative impact.
DISTRIBUTION OF NEUROPEPTIDE Y NEURONS IN THE MALE NATIVE THAI CHICKEN

B. Kamkrathok\*(1), Y.Chaiseha(1)

(1) Institute of Science, Suranaree University of Technology, Nakhon ratchasima, Thailand

* Corresponding author: ko_boonyarit@hotmail.co.th

Neuropeptide Y (NPY), a 36 amino acid neurotransmitter, plays various physiological functions involving food intake in birds. NPY is associated with the regulation of the reproductive system in the female native Thai chickens. To date, there are no data described the functional aspects of NPY in the male birds. Thus, the objective of this study was to elucidate the distributions of NPY-immunoreactive (-ir) neurons and fibers in the male brain, enabling further studies of neuroendocrinology related to feeding behavior of the male native Thai chicken. Six male native Thai chickens, Pradoohangdum breed, were used and the distributions of NPY-ir neurons and fibers were detected by immunohistochemistry technique. The results revealed that the distributions of NPY-ir neurons and fibers were located throughout the brain, especially in the hypothalamus. The greatest density of NPY-ir neurons was found within the nucleus septalis lateralis and nucleus paraventricularis magnocellularis(PVN). A few of NPY-ir neurons was observed within the nucleus inferioris hypothali (IH), and the nucleus infundibuli hypothali (IN). The distribution of NPY-ir fibers was found in abundance within the nucleus supraopticus, pars ventralis, nucleus suprachiasmaticus, pars medialis, IH-IN, PVN, and the external layer of eminentia mediana (median eminence). A few of NPY-ir fibers was observed within the nucleus commissurae pallii, nucleus periventricularis hypothalamic, commissura pallii, regio lateralis hypoyhalami, and nucleus rotundus. These results indicated the distributions of NPY-ir neurons and fibers in the male native Thai chicken were markedly observed in the PVN which might be related to the regulation of feeding behavior in this equatorial species.
ID: 681

GROWTH PERFORMANCE, HAEMATOLOGICAL AND BIOCHEMICAL PARAMETERS CHANGES IN BROILERS CHICKENS FED VARYING LEVELS OF VERNONIA AMYGDALINA LEAF MEAL

M. Tokofai\*(1), K.Togbenou(2), O.N'nanle(3), K.Idoh(2), A.Agbonon(2), M.Gbeassor(2)

(1)CERSA / University of Lome, Lome, Togo

(2) University of Lome, Lome, Togo

(3) University of Lome / CERSA, Lome, Togo

* Corresponding author: marctokofai@gmail.com

The growing concerns on the use of antibiotics as growth promoters in animal diets and its health implication on consumers of poultry products has necessitated a search for suitable alternative growth promoting agents especially medicinal plants. The aim of this study was to determine the effects of dietary supplementation of Vernonia amygdalina leaf meal (VALM) on the haematology, biochemical and growth performance of broilers chickens. Four hundred and eighty day-old cobb chicks were randomly allocated to four dietary treatment viz.: a basal diet (V0) (control group with 0% of VALM); basal diets with 1 (V1), 2 (V2) and 3% VALM (V3) having 4 replicates of 30 birds each in Completely Randomized Design. Birds were managed conventionally on a deep litter system for a period of 6 weeks. Data were collected on body weights, feed intake, feed conversion ratio and blood parameters. Results showed that feed intake of the birds in V2 and V3 VALM was significantly lower (P0.05) but significantly higher than those of V2 and V3. Final body weights and weight gain were higher significantly (PVernonia amygdalina leaf supplementation improved growth performance, haematological and biochemical parameters indices of broiler chickens.

References

Alaraa R., H. Abdurahmana, S. K. A. Mudalipa (2017). "Phytochemical and pharmacological properties of Vernonia amygdalina: a review." *80*: 96.

Mohammed, A. and A. Zakariya'u (2012). "Bitter leaf (Vernonia amygdalina) as a feed additive in broiler diets." *6*(3): 38-41.
EFFECT OF SUPPLEMENTATION OF LYSOLECITHIN ON PRODUCTION PERFORMANCE AND INTESTINAL MORPHOLOGY OF CHICKEN

E. Subbiah*(1), C. Pandian(1), S.T. Selvan(1), N.Karthikeyan (1), A.V.Omprakash(1)

(1) TAMIL NADU VETERINARY AND ANIMAL SCIENCES UNIVERSITY (TANUVAS), INDIA, Chennai, India

* Corresponding author: vetezhil2015@gmail.com

Lysolecithins are produced by an enzymatic conversion of lecithin. The addition of lysolecithin to diet decreased the size of fat globules and increased the active surface of fats for enzymatic digestion. Hence, the present research was conducted to study the effect of lysolecithin on production performance and intestinal morphology of Nandanam broiler 3 at Poultry Research Station, Tamil Nadu Veterinary and Animal Sciences University, Chennai, India. A total of nine hundred day old, unsexed, Nandanam broiler 3 chickens were wing banded, weighed and randomly allotted into three treatment groups and reared under deep litter system of management. All the treatment had three replicates and each consisted of 100 chicks. Birds were reared upto eight weeks of age. The experimental birds fed with diets containing lysolecithin 0 g (T1-control), 2.5 g (T2) and 5 g (T3). All the diets were iso-caloric and iso-nitrogenous. Birds were fed with pre-starter (0-14 days), starter (15-28 days) and finisher (29-56 days), respectively. The body weight, body weight gain, feed consumption, feed efficiency, livability, NewYork dressed weight, eviscerated carcass weight, ready - to - cook yield and economics were analyzed. The data collected on various parameters were grouped and subjected to statistical analysis of one way ANOVA as per the procedure of statistical analysis system (SPSS, version 20.0 for windows). Dietary incorporation of 5 g of lysolecithin (T3) recorded significantly (P≤0.01) higher body weight followed by 2.5 g of lysolecithin (T2) and control (T1) group at 8 weeks of age. The highly significant (P<0.01) difference in body weight gain of Nandanam broiler-3 was observed in T3 group from second weeks to entire experimental period. Feed consumption revealed significant (P<0.05) difference between treatment groups throughout the experiment period. Birds received 5 g of lysolecithin (T3) had better (P<0.01) feed efficiency. Statistical analysis revealed a significant (P<0.05) difference in New-York dressed weight, eviscerated carcass weight, ready - to - cook yield between treatment groups. Feeding of 5 g of lysolecithin to Nandanam broiler 3 significantly (P<0.01) increased intestinal length when compared to other treatment groups. The cost effectiveness of T3 group showed increased net profit per kg live weight. From the experiment, it was concluded that 5 g of lysolecithin can be incorporated in the feed for better returns to the farmers.
EFFECT OF EARLY THERMAL CONDITIONING ON THE HEPATIC METABOLISM IN BROILER CHICKS

Y. Ouchi\*(1), V. Chowdhury(2), T. Bungo(3)

(1) Hiroshima University, Higashihiroshima, Japan

(2) Laboratory of Stress Physiology and Metabolism, Division of Experimental Natural Science, Faculty of Arts and Science, Graduate school of Bioresource and Bioenvironmental Science, Kyushu University, Fukuoka, Japan

(3) Laboratory of Animal Behavior and Physiology, Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi hiroshima, Japan

* Corresponding author: d190610@hiroshima-u.ac.jp

**【Objectives】** The metabolic heat production is one of the main heat production systems in chickens, and the liver is an important heat producing tissue in the system of metabolic heat production. Besides, it is well known that thermal conditioning on early age improves post-growth thermotolerance and alters heat production systems in chickens. The purpose of this study is to clarify the effect of thermal conditioning at early age on metabolism of liver in broiler chicks. **【Materials & Methods】** Three-day-old chicks were exposed at 40°C for 12 hours (thermal conditioning) and control chicks were kept at 30°C. At one week old, the both groups were exposed 40°C for 15 minutes (heat challenge), or kept at 30°C. In this study, there is 4 groups depending on whether there is thermal conditioning and whether there is heat challenge. After heat challenge, the all groups of liver tissues were collected for metabolomics analysis. **【Results】** Results of metabolomics analysis, the level of isoflavones such as daidzein, genistein and glycitein were low by thermal conditioning. In addition, the effect of heat challenge was observed in the cholesterol-related metabolites. **【Conclusion】** In conclusion, thermal conditioning at early age alters the part of metabolism of liver in broiler chicks.
IMPACT OF REPLACING CHOLINE CHLORIDE WITH A PHYTOBIOTIC ON THE HISTOMORPHOMETRIC
CHANGES IN THE LIVER OF LAYING HENS

J. Gonzalez(1), H.Morales*(2), L.Rivera(1)

(1)Nuproxa Switzerland Ltd., Etoy, Switzerland

(2)DVM, MSc, External Consultant, Santiago de cali, Colombia

* Corresponding author: javier.gonzalez@nuproxa.ch

The use of phytobiotics as an alternative to products of chemical synthesis in animal production is increasing due to environmental and consumer demands. A phytobiotic-based product, Biocholine® (Nuproxa, Switzerland Ltd.; Indian Herbs Specialities PVT. Ltd.) composed of the medicinal plants Andrographis paniculata, Azadirachta indica, Trachyspermum ammi and Achyranthes aspera, with proven lipotropic activity, has been successfully used to replace choline chloride (CC) in broiler feeds. The objective of this study was to compare the dietary supplementation of CC vs Biocholine® on the histomorphometric changes in the liver of layers during production. The experiment consisted of two dietary treatments; a corn and soybean meal-based diet supplemented with CC 60% (800 g/ton) (T1) and T1 but supplemented with 200 g/ton of Biocholine® in replacement of CC (T2). A total of 53.677 layers (Shaver Brown) were distributed in 4 houses with floor production, automatic chain feeders and bell drinkers. The layers allocated in houses 1 and 2 (n = 26.928) were fed T1 and the ones allocated in houses 3 and 4 (n = 26.749) were fed T2. The experimental period was 20 weeks (34 to 54 wk of age). The study was conducted in a layer farm, located at the Santander Department, Colombia, at an altitude of 1.055 m over sea level. At 38, 42, 46, 50 and 54 wk of age, 5 layers from each house and dietary treatment were randomly sampled for liver samples (n = 60 in each treatment). Liver samples were submitted to histomorphometric analysis. Although not relevant for the objectives of the study, egg production was recorded. Data across ages were submitted to an analysis of means and variances for parametric values using the Snedecor F distribution for comparing the variances as reported by Gibson-Corley et al. (2013). Results showed that CC induced a more extensive and severe cellular degeneration in liver (p 0,05) were observed in the severity or extension of inflammatory-related damage in liver tissue. Egg production and laying persistency were improved (p vs CC. In conclusion, the use of a phytobiotic product based on medicinal herbs proved to be an alternative to substitute CC in the feed of laying hens.
EFFECT OF FERMENTED RAPESEED CAKE ON PHOSPHORUS DIGESTIBILITY IN LAYING HENS

A. Drażbo\*(1), M.Kopacz(1), K.Kozłowski(1)

(1)University of Warmia and Mazury in Olsztyn, Poland, Olsztyn, Poland

* Corresponding author: aleksandra.drazbo@uwm.edu.pl

Fermentation is considered to be an effective method for improving the nutritional value of diets for farm animals. Fermentation increases the bioavailability of selected nutrients and decreases the content of undesirable compounds in many feedstuffs. The aim of this study was to determine the effect of fermentation on the phytate-phosphorus content of rapeseed cake as well as ileal and total tract phosphorus (P) digestibility in laying hens. A total of 216 Hy-Line Brown layers at 36 weeks of age, kept in battery cages, were divided into 3 dietary treatment groups (36 replicates of 2 birds each). For 12 weeks, the birds were fed isonitrogenous and isocaloric diets. In the control group, the main dietary protein source was soybean meal (C), and the diets in two experimental groups were supplemented with 20% of raw rapeseed cake (RRC) or fermented rapeseed cake (FRC). During the experiment, excreta were collected for 5 days to determine total tract P digestibility. On the last day of the experiment, 10 hens from each group were sacrificed and the small intestinal digesta was collected to evaluate ileal P digestibility. TiO2 was used as an indigestible dietary marker The differences between group means were determined by ANOVA and Tukey's test with the use of Statistica 13.1 software. It was found that fermentation led to a nearly two-fold decrease in the phytate-phosphorus content of rapeseed cake (7.88 g/kg vs. 14.28 g/kg). Ileal P digestibility was significantly higher in hens fed FRC than in those receiving RRC, and comparable values were noted in FRC and control groups (FRC - 63.9%, RRC - 50.9%, C - 62.1%, P≤0.001). Fermentation had no positive effect on total tract P digestibility (P>0.05). The coefficients of total tract P digestibility were considerably lower in all groups (≤ 30%), probably because the absorbed P was re-directed into the excreta via the urine. It can be concluded that fermentation decreases the phytate-phosphorus content of rapeseed cake and exerts a beneficial influence on ileal P digestibility.