ID: 1001

# EFFECTS OF HEAT STRESS ON RECENTLY ESTABLISHED PRIMARY HEPATOCYTE - NON-PARENCHYMAL CELL MONO- AND CO-CULTURE MODELS OF CHICKEN ORIGIN

M.Mackei*(1), S.Nagy(2), A.Molnár(2), F.Husvéth(2), K.Dublecz(2), Z.Neogrády(1), G.Mátis(1)

(1) Division of Biochemistry, Department of Physiology and Biochemistry, University of Veterinary Medicine, Budapest, Hungary, (2) Department of Animal Sciences, Georgikon Faculty, University of Pannonia, Keszthely, Hungary

*Presenting author

**Corresponding author mail:** mackei.mate@univet.hu (Máté Mackei)

Heat stress is one of the most important environmental factors in broilers affecting both animal health and productivity. The major aim of this study was to establish novel avian hepatocyte - non-parenchymal cell co-culture models for investigating the cellular effects of heat stress and its interaction with inflammation in chicken liver.

In order to isolate the different cell types, the liver of 21-day-old Ross 308 broilers was flushed using multi-step perfusion and digested by collagenase supplemented medium, followed by differential centrifugation. Hepatocyte mono-cultures and hepatocyte - non-parenchymal cell (predominantly Kupffer cells) co-cultures (with cell ratio of 6:1) were prepared, the latter mimicking a moderate inflammatory state. Isolated and cultured cells were characterized by immunocytochemistry and flow cytometry. Cells were cultured as confluent monolayers either at 38.5oC (control) or were exposed to 43oC (heat stress) for 1 or 2 h. CCK-8 assay was used to investigate the cellular metabolic activity. The concentration of extracellular H2O2 was measured by Amplex Red method, while heat shock protein 70 (HSP70), interleukin- (IL)-6 and IL-8 concentrations of culture media were determined by chicken specific ELISA kits. Differences between the groups were assessed using one-way analysis of variance (ANOVA) and post-hoc test.

Shorter, 1 h long heat exposure significantly increased metabolic activity and extracellular H2O2 release, while HSP70, IL-6 and IL-8 production were significantly decreased, compared to controls on both cell culture models. In spite of this, all the alterations were normalized after 2 h heat stress, assuming an adaptation and restoration of liver cells. Mono-cultures and co-cultures responded in a similar manner to heat exposure.

In conclusion, the newly developed primary cell culture models provide suitable tools for studying the hepatic inflammatory and stress response. The results of this investigation highlight the effects of short term heat stress on the different liver-derived cells, suggest the specific role of oxidative stress in acute stress response and suppose an efficient cellular adaptation mechanism in liver cells.

The study was supported by the NKFIH grant no. 124586 of the Hungarian National Research, Development and Innovation Office.
ID: 1004

# INVESTIGATION OF THE EFFECTS OF T-2 TOXIN ON CO-CULTURED PRIMARY CELLS FROM CHICKEN LIVER

M.Mackei*(1), A.Molnár(2), F.Husvéth(2), K.Dublecz(2), Z.Neogrády(1), G.Mátis(1), H.Fébel*(3)

(1) Division of Biochemistry, Department of Physiology and Biochemistry, University of Veterinary Medicine, Budapest, Hungary, (2) Department of Animal Sciences, Georgikon Faculty, University of Pannonia, Keszthely, Hungary, (3) Research Institute for Animal Breeding, Nutrition and Meat Science, National Agricultural Research and Innovation Centre, Herceghalom, Hungary

*Presenting author

Corresponding author mail: mackei.mate@univet.hu (Máté Mackei)

Mycotoxin contamination of animal feed has a high importance in animal husbandry worldwide. T-2 toxin is able to affect cellular metabolic processes and to cause oxidative stress as well as it has an immunomodulatory activity. The hypothesis of our study was, that T-2 toxin may alter cellular metabolic activity, hydrogen peroxide (H2O2) and pro-inflammatory cytokine production of primary liver-derived cell culture models of chicken origin.

Various cell types were isolated by differential centrifugation from a freshly perfused chicken liver. Hepatocyte mono-cultures and hepatocyte - non-parenchymal cell (predominantly Kupffer cells) co-cultures (with cell ratio 6:1) were prepared. After 24 h culturing, cells were replenished with fresh medium with or without T-2 toxin (0, 10, 100, 1000 nmol/l) for 8 and 24 h. The metabolic activity, reactive oxygen species (extracellular H2O2) production, the concentration of heat shock protein 70 (HSP70) and interleukin (IL-)6 and IL-8 in culture media were assessed. Differences between the groups were evaluated using one-way analysis of variance (ANOVA) and post-hoc test. Results were considered significant in case of p < 0.05.

T-2 toxin treatment significantly decreased catabolic metabolism in all of the applied concentrations after both incubation times in every applied model. Concentrations of HSP70 and IL-8 were significantly increased in hepatocyte mono-cultures treated with 100 and 1000 nmol/l T-2 toxin containing culture medium after 24 h incubation. Similarly, IL-6 levels were also significantly elevated in the 100 and 1000 nmol/l groups in both of mono- and co-cultures, but only after 8 h of incubation time. In spite of the mentioned effects of T-2 toxin treatment, no significant differences were observed in reactive oxygen species production. Regarding the cell type specific differences, independently of the T-2 toxin challenge, the applied models showed different levels of H2O2, HSP70 and IL-8 concentrations.

In conclusion, the established primary chicken-derived hepatic cell culture models proved to be suitable for study the cellular effects of T-2 toxin. Metabolic activity and immune status of the cell cultures were intensively affected, but no changes were found in H2O2 levels at the same.

The study was supported by the EFOP-3.6.3-VEKOP-16-2017-00005 grant of the Hungarian Ministry of Human Resources and by the NKFIH grant no.124586 of the Hungarian National Research, Development and Innovation Office.
ID: 1139

# GHR GENE CONTROLS MITOCHONDRIAL FUNCTION BY REGULATING MITOCHONDRIAL BIOGENESIS VIA IGF1 SIGNALING IN CHICKEN

B.Hu\*(1), C.Zhao(2), H.Wei(2), H.Li(3), X.Zhang(3)

(1) SCAU, Guangzhou, China, (2) na, Guangzhou, China, (3) College of Animal Science, South China Agricultural University, Guangzhou, China

\*Presenting author

**Corresponding author mail:** 1012386229@qq.com (Bowen Hu)

Mitochondrial biogenesis is pivotal for its function, which is defined as a sophisticated and dynamic process that controls mitochondria renew, content as well as quantity, including mitochondrial DNA (mtDNA) transcription and translation, translation of nucleus-derived transcripts, recruitment of newly synthesized proteins, etc [1]. Our previous study has revealed that mitochondrial dysfunction in sex-linked dwarf (SLD) chickens' skeletal muscle, indicating that GHR gene is essential for chicken mitochondrial function [2]. However, the underlying molecular mechanism remains unknown.

In this study, we overexpressed or interfered with the expression of the GHR gene together with IGF1 gene in chicken primary myoblast cells to investigate their role on mitochondrial biogenesis and function. Mitochondrial biogenesis is assessed by Mito-Tracker Green (MTG) staining, the expression of mitochondrial biogenesis markers, the level of DNA transcription as well as mtDNA content. Mitochondrial function is assessed by mitochondrial membrane potential (6Wm) and ATP level. The data were presented as means ± standard error of the mean (SEM), and the statistical analyses were performed using Student's t-test and the significance was represented by p-values, p < 0.05 was considered to be statistically significant.

The results showed that overexpression of the GHR gene significantly promoted mitochondrial biogenesis and function, as indicating by elevated fluorescence intensity of MTG staining, the expression of mitochondrial biogenesis markers (PGC1A, PGC1B, PRC, NRF1, NRF2, TFAM), the level of DNA transcription (ND1, CYTB, COX1, ATP6) and mtDNA content, as well as the 6Wm and ATP level. Conversely, its loss of function repressed mitochondrial biogenesis and function. Similar consequences were also observed after overexpression or knockdown of the IGF1 gene. Additionally, overexpression of IGF1 gene attenuated the effects of knockdown of the GHR gene on mitochondrial biogenesis and function. Taken together, we corroborated that the GHR gene controls mitochondrial function by regulating mitochondrial biogenesis via IGF1 signaling in chicken.

## References

1. Zhu J, Wang KZ, Chu CT. After the banquet: mitochondrial biogenesis, mitophagy, and cell survival. AUTOPHAGY 2013, 9 (11).

2. Hu B, Hu S, Yang M et al. Growth hormone receptor gene is essential for chicken mitochondrial function in vivo and in vitro. INT J MOL SCI 2019, 20(7).
ID: 1277

COMPARATIVE ASSESSMENT OF IMMUNE RESPONSE GENES IN CHICKEN ACETYLCO-A CARBOXYLASE A (ACACA) GENE KNOCKDOWN PRIMARY LIVER CELL CULTURE

A.Prasad*(1), B.T.k.(2), C.R.n.(2), G.Neerasa(3), P.Kumar(4), B.Bhushan(4), C.Paswan(2), S.G(5), T.J(6)

(1) ICAR-Directorate of Poultry Research, Mancherial, India, (2) ICAR-Directorate of Poultry Research, Hyderabad, India, (3) Department of Animal Husbandry, Andhra Pradesh, India, (4) ICAR-Indian Veterinary Research Institute, Bareilly, India, (5) PVNR Telangana Veterinary University, Hyderabad 500030, Telangana, India, Hyderabad, India, (6) PVNR Telangana Veterinary University, Hyderabad, India

*Presenting author

Corresponding author mail: rajendraprasad.vet@gmail.com (Athe Rajendra Prasad)

The Acetyl-CoA carboxylase A (ACACA) is a rate-limiting enzyme which catalyzes the carboxylation of acetyl-CoA to produce malonyl-CoA in liver. The objective of the study is to explore the profile of immune response genes in the knock-down chicken liver cells. The knock down liver cells were developed with two shRNA molecules for ACACA gene and were available in the laboratory to study the differential expression levels of immune response genes in knock-down and control chicken hepatocytes. Quantitative real time PCR was performed to investigate the expression levels of two immune response genes namely, Interferon Alpha (IFNA) and Interferon Beta (IFNB) in knock-down and control hepatic cell culture. The statistical analysis (One way ANOVA) was carried out using SPSS 25 version. The gene expression levels (40-6Ct) of IFNA were 35.28 in shRNA1 knock-down and 35.61 in shRNA2 knock-down hepatocytes. In contrast to control, IFNA expression showed an up-regulation of 1.01 and 1.26 folds in shRNA1 and 2 knock-down cells, respectively. The IFNB expression showed a down-regulation of 1.82 fold in shRNA1 knock-down and 1.32 fold up regulation in shRNA2 knock-down cells, respectively. There was non-significant (P>0.05) difference of expression of both IFNA and IFNB genes between knock-down and control cells. From these results, it is concluded that the potential knock-down cells for ACACA gene revealed non-significant effect on expression of immune response genes under in vitro cell culture system. These shRNA molecules may be used further for developing knock-down chicken to analyze the effect knock down on physiological and immunological system in animals.

Key words: shRNA, knock down, ACACA, liver cells, chicken, immune response genes
ID: 1282

# THE INFLUENCE OF DIFFERENT FRACTION OF ALUMINOSILICATE IN SLAUGHTER CHICKEN DIET ON JEJUNUM MORPHOLOGY

J.Bogucka*(1), M.Banaszak(2), A.Dankowiakowska(1), J.Biesek(2), B.Bigorowski(2), A.Zmudzinska(2)

(1) Department of Physiology, Zoophysiotherapy and Animal Nutrition, Faculty of Animal Breeding and Biology, UTP - University of Science and Technology, Bydgoszcz, Poland, (2) Department of Animal Breeding, Faculty of Animal Breeding and Biology, UTP - University of Science and Technology, Bydgoszcz, Poland

*Presenting author

Corresponding author mail: bogucka@utp.edu.pl (Joanna Bogucka)

The production of broiler chickens pays attention to high emission of toxic gases as the ammonia, hydrogen sulfate and carbon dioxide to the atmosphere. Searching of solutions a.e.: feed supplements for decreasing toxic gases emission level by the higher absorption capacity in digestive tract. The aim of this study was compare feed supplementation by different fractions of aluminosilicate (zeolite, halloysite) and evaluation of impact on slaughter chickens Ross 308 digestive tract morphology. In this study 3 groups of chickens Ross 308 (100 bird per group) were evaluated. In all groups birds were fed with standard diet. In group I (control) fed without aluminosilicate was used. The group II was fed with 1% halloysite additive. In the group III fed with 1% zeolite addictive was used. After 6 weeks slaughtered 30 chickens (10 bird per trial group). After slaughtering small intestine samples were collected (3-4 mm long) for histological analysis. In samples detected: villus height and widgt, which were used to villus surface area and crypt depth calculation. Statistical significance difference were found in villus height between group with 1% halloysite additive (II) and group with 1% zeolite additive (III) in fed. Similar differences were found between group I and III. No significant differences were found in the villus height between group I and II. The statistical significant difference of villus widgt was found between groups I and III, II and III as well. High significance differences (P<0.01) of villus surface area were prove for group with halloysite additive compared to group with zeolite additive in feed. Between control group and group with zeolite the statistical significant difference was also detected. Groups I and II were no significant different accordance mentioned traits. Smaller crypt depth in digestive tract od birds from group III was detected compare birds from groups I and II. These differences were statistical significant (P<0.05). Additive of 1% of halloysite to feed improve quality of villus evaluated on villus height, villus widgt, villus surface area and crypt depth of small intestine.

<footnote>*This work has been supported by the European Agricultural Foundations for Rural Area Development " PROW 2014-2020, "Cooperation – Safe Farm – Product, Process and Marketing Innovation in Broiler Chicken Production"</footnote>
Hygiene and pathology
ID: 15

ACUTE PHASE PROTEIN AS BIOMARKERS TO DETECT INNATE IMMUNE
RESPONSES TO THE INFLAMMATION TRIGGERED BY ESCHERICHIA.
COLI LIPOPOLYSACCHARIDE IN BROILER CHICKENS

H.Kaab\*(1), M.Kaab(2), D.Eckersall(2)

(1) University of Kufa, Glasgow, United kingdom, (2) University of Glasgow, Glasgow, United states
\*Presenting author

**Corresponding author mail:** haidert.mehde@uokufa.edu.iq (Haider Kaab)

Lipopolysaccharides of Gram-negative bacteria can induce an APP response and can cause inflammation sickness and decreased production in broiler chickens (Mireles, et al., 2005; Xie, et al., 2000). Chickens are one of the important avian biomedical models that have provided important physiological, anatomical and molecular insights for immunology (Packialakshmi, et al., 2016). The aim of the study is to determine if monitoring APP can provide biomarkers of the innate immune response to LPS and related diseases and their association with systemic immune changes in established biomarkers for early diseases detection.

Inflammation was triggered in broiler chickens (15-day old) by subcutaneous injection of 2mg of LPS of E. coli per kg body weight in order. The APR was monitored by using ELISA for a-1 acid glycoprotein (AGP) and serum amyloid A (SAA) and radial immune-diffusion for estimation of ovotransferrin (OVT) concentration in plasma samples at 0, 6, 12, 24, 48 and 72 h post treatment. A digital thermometer was used to record rectal temperature at the time of blood sampling.

Acute phase response triggered by E. coli LPS revealed significant changes for AGP which was significantly higher in the LPS treated group on 12h post treatment (P<0.001) with 2.7-fold increase from 0.68 g/L (0.37-1.6) median (range) in control group to 1.84 g/L (0.9-2.6) in the LPS group with this difference remaining significant till 72h. SAA levels were significantly higher (P<0.001) in LPS group on 6h post treatment with a 128.8-fold, increase from 21.3 µg/ml (12.6-56.1) in control group to 2744 µg/ml (1346.9-3817.3) in LPS group. Then the difference decreased to be non-significant on 72h post treatment. OVT became higher significantly (P<0.05) in LPS group on 12h post treatment with 1.72-fold, from 0.98 mg/ml (0.89-1.32) median (range) in control group to 1.69 mg/ml (1.39-2.06) and remained significantly higher in the LPS group till 72h PT. Pyrexia triggered by LPS showed significant increase in body temperature estimated by rectal measurement from 41.1C (41-41.6) median (range) in control group to 41.7C (41.2-42.3) in the LPS group. The difference remains significant on 12 h P.T. then decreased to be not significant thereafter.

In conclusion, E. coli LPS induced high level of SAA, AGP and OVT. The highest and earliest response was the SAA after 6h. These proteins could be use as biomarkers for diagnosis and prognosis of the diseases.
ID: 570

THE INCREASE OF EIMERIA INFECTION REGULATED OXIDATIVE STRESS
IN BROILER CHICKEN AND ITS STRONG CORRELATION WITH GROWTH
PERFORMANCE AND GUT HEALTH PARAMETERS

P.Teng\*(1), Y.Tompkins\*(2), S.Yadav\*(1), F.Castro\*(1), A.Fuller\*(1), W.Kim\*(1)

(1) University of Georgia, Athens, ga, United states, (2) University of Goergia, Athens, ga, United states
\*Presenting author

Corresponding author mail: pt00221@uga.edu (Po-Yun Teng)

Coccidiosis is a parasitic disease caused by genus Eimeria which induces malnutrition, gut damage, and high mortality. Our previous study has shown that the higher Eimeria challenge doses linearly regulated gut health parameters, including apparent ileal digestibility, gut permeability, and gut morphology. In the present study, further analysis was conducted to evaluate the effects of graded Eimeria challenge doses on gene expression of tight junctions and parameters of oxidative stress in the liver and serum. The correlation coefficients among doses, growth performance, and several parameters were calculated to understand the relationship among severity of Eimeria infection, gut health, and oxidative stress in broilers. The study includes control group and four other treatments with different levels of Eimeria doses. A mixed Eimeria spp. solution with 50,000 E. maxima, 50,000 E. tenella, and 250,000 E. acervulina per milliliter was prepared for the highest dose challenge treatment. The two-fold serial dilution was used to make the medium-high (25,000 E. maxima; 25,000 E. tenella; 125,000 E. acervulina), the medium-low (12,500 E. maxima; 12,500 E. tenella; 62,500 E. acervulina), and the lowest challenge dose (6,250 E. maxima; 6,250 E. tenella; 31,250 E. acervulina). A total of 360 thirteen-day-old male broilers were randomly allocated into five treatments with six replicated cages and challenged on the same day. The mucosa of jejunum and liver were collected on 6-d post-infection. The results showed that the gene expression of tight junction protein was linearly up-regulated with the increase in challenge doses. Moreover, Eimeria challenge increased superoxide dismutase (SOD) and decreased glutathione in the liver. However, only the lowest challenge dose upregulated gene expression of catalase (CAT) in the liver and serum. SOD and CAT did not linearly increase in response to the graded challenge doses. The results suggested that the severe Eimeria infection might damage and suppress the mechanisms of antioxidative systems in the liver of broilers. Furthermore, strong linear relationships were found between challenged doses and growth performance (r = -0.911), as well as the villi height to crypt depth ratio in the jejunum (r = -0.789). In conclusion, Eimeria infection increased oxidative stress in the liver of broilers. Moreover, the increase of Eimeria challenge was strongly correlated with growth performance and parameters of intestinal integrity.
ID: 723

# EPIDEMIOLOGICAL INVESTIGATION OF MYCOPLASMA GALLISEPTICUM AND MYCOPLASMA SYNOVIAE PREVALENCE IN COMMERCIAL LAYER AND LAYER BREEDER FLOCKS IN GREECE.

D.Koutsianos*(1), L.Athanasiou(2), D.Mossialos(3), K.Koutoulis(4)

(1) DEPARTMENT OF POULTRY DISEASES, FACULTY OF VETERINARY MEDICINE, UNIVERSITY OF THESSALY, KARDITSA, GREECE, Athens, Greece, (2) Clinic of Medicine, Faculty of Veterinary Science, University of Thessaly, Karditsa, Greece, Karditsa, Greece, (3) Department of Biochemistry and Biotechnology, University of Thessaly, Larissa, Greece, Larissa, Greece, (4) Department of Poultry Diseases Faculty of Veterinary Science, University of Thessaly, Karditsa, Greece, Karditsa, Greece

*Presenting author

**Corresponding author mail:** dkoutsianos@yahoo.gr (Dimitrios Koutsianos)

Mycoplasma gallisepticum and synoviae are important pathogens for poultry production. Mycoplasma gallisepticum is known to cause inflammation of the air sacks and chronic respiratory disease having also an effect on egg production, while Mycoplasma synoviae is mainly related to arthritis, respiratory infection and egg quality problems.

The scope of the present survey was to investigate the prevalence of Mycoplasma gallisepticum and Mycoplasma synoviae in commercial layer and layer breeder flocks of various ages and areas in Greece. For this purpose, 61 different flocks were sampled (4 layer breeder flocks and 57 commercial pullets-layer flocks) during the period 2016-2017.

Flocks appeared to have a normal or increased mortality status. Ten serum samples were collected from each flock in order to be tested with the Enzyme-Linked Immunosorbent Assay (Synbiotics Elisa Proflok MG and Synbiotics Proflok Ms kits). When samples were negative, a second sampling was done in 3 to 6weeks time to check for seroconversion. None of the selected flocks was vaccinated against MG or MS; therefore the presence of positive antibody titers was indicating exposure to mycoplasma. At least 20% of positive samples out of the 10 samples collected from each flock was needed to characterize a flock as 'infected'. (OIE terrestrial manual, 2018). None of the 4 layer breeder flocks was found to be positive for MG while high prevalence 86% (37/43 flocks) was observed for commercial layers and 64% (9/14 flocks) for rearing pullets. Forty two out of 46 positive flocks appeared to have 100% positivity in serum samples (10/10) while 2 flocks had 90% positivity, one flock 60% and 1 flock 30%. For Mycoplasma synoviae a markedly higher level of prevalence was revealed. All commercial layer flocks (43/43) and rearing pullet flocks (14/14) were positive, while no infection was reported for the layer breeder flocks (0/4 flocks). Concerning MS positive flocks, 56 out of 57 flocks had 100% positive serum samples.

In conclusion, our study reported a high prevalence of Mycoplasma gallisepticum and Mycoplasma synoviae in commercial layer and rearing pullet flocks while no exposure to the pathogens was observed for the tested layer breeder flocks.

Reference: OIE terrestrial Manual, Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, 8th Edition, 2018, Volume, chapter 3.3.5, Avian mycoplasmosis
ID: 751

# INFECTIOUS LARYNGOTRACHEITIS HVT-ILT AND HVT-ND-ILT CONSTRUCTS: REPLICATION AND EFFICACY AGAINST CHALLENGE IN SPF BIRDS.

I.Alvarado*(1), M.Garcia(2), D.Maekawa(2), A.Mendoza-reilley(3)

(1) Merck Animal Health, Watkinsville, United states, (2) University of Georgia, Athens, United states,

(3) Merck Animal Health, Athens, United states

*Presenting author

**Corresponding author mail:** ivan.alvarado@merck.com (Ivan Alvarado)

The capacity of two HVT based constructs, HVT-ILT (Innovax®-ILT) and HVT-ND-ILT (Innovax ND-ILT®), to protect SPF birds against challenge with a virulent infectious laryngotracheitis strain was evaluated. Additionally, the benefits of administering a live attenuated vaccine to HVT-ILT vaccinated birds on further reduction of clinical signs and viral shedding post-challenge were assessed.

Design. The HVT-ILT (Innovax®-ILT), HVT-ND-ILT (Innovax ND-ILT®) and attenuated B6ORFC (ORF C gene deleted developed by Dr. Maricarmen Garcia, University of Georgia) vaccines were administered in ovo at 19 days of embryonation. One hundred and seventy-five SPF chickens were equally allocated to the following groups: a) HVT-ILT vaccinated ; b) HVT-ND-ILT vaccinated ; c) HVT-ILT & B6ORFC vaccinated; d) non-vaccinated/Challenged; e) Non-vaccinated/Non-challenged. At 1-, 2-, 3-, and 4-weeks post vaccination feather pulps were processed for DNA extraction and HVT real-time PCR.

At 28 days of age, chickens from groups a, b, c and d were challenged with 10^3.8 TCID50 of the ILTV virulent strain 1874c5. Chickens were monitored for clinical signs from day 2 to 7 post-challenge. Tracheal swabs were collected at 3- and 5-days post challenge to determine the viral genome load in the trachea. To be considered protected, vaccinated chickens should show significant decreases in clinical signs and challenge virus genome load in the trachea as compared to the non-vaccinated/challenged group.

Both, HVT-ILT and HVT-ND-ILT vaccines were detected in feather pulps in 100% of the evaluated birds at 7, 14 and 21 days, decreasing to 60% and 50% at 28 days, respectively. No significant differences in HVT genome loads were observed in feather follicles between the HVT-ILT and HVT-ND-ILT vaccinated groups. Both vaccines provided protection against challenge with no significant differences in clinical signs or viral genome loads in tracheal swabs post-challenge. In contrast, significant differences (p< 0.05) in clinical signs and viral genome loads post-challenge were observed between HVT-ILT and HVT-ND-ILT vaccinated groups and the non-vaccinated/challenged group. HVT-ILT vaccinated birds that received the live attenuated B6ORFC strain not only showed a significant reduction in clinical signs but also exhibited the highest reduction in the level of replication of the virulent ILTV post-challenge.
EVALUATING THE IMPACT OF IN OVO INJECTED LACTOBACILLUS ON BROILER WELL BEING

C.Catañeda*(1), L.Zhang(1), K.Wamsley(1), C.Mcdaniel(1), M.Beck(2), A.Kiess(1)

(1) Mississippi State University, Mississippi state, United states, (2) Mississippi State Unversity, Mississippi state, United states

*Presenting author

Corresponding author mail: cdc748@msstate.edu (Claudia Castaneda)

Lactobacillus, a probiotic bacterium, is known to modulate GABA receptor expression related to stress and immune function in mice. Therefore, it is hypothesized that an early administration of Lactobacillus to broilers could result in stress and immune function modulation improving broiler well-being (welfare and health). Hence, the objective of this study was to determine if the injection of different Lactobacillus strains into fertile broiler eggs impacts GABA expression and immune parameters. On d18 of incubation, 4 in ovo treatments (T) were applied: T1= Marek's vaccine (MV) with no probiotic, T2= MV+ L. animalis, T3= MV+ L. reuteri, T4= MV+ L. rhamnosus, all at ~10^6 cfu/50µl. Injected eggs were transferred to 3 hatchers per T. At hatch, 720 male chicks were moved to a grow-out facility for 42d, placing 18 chicks per pen, with a total of 10 pens/T. A coccidiosis challenge was performed on d14 to induce stress. On d0, 14, 21, 28 and 42, spleen, bursa, brain and blood samples were collected. Spleen and bursa were weighed and bursa follicular area (BFA) was measured. Cytokine (blood) and GABA (brains) expression were quantified utilizing ELISA kits. Data were analyzed using a randomized complete block design and means were separated using fishers protected LSD when Pc0.05. For immune parameters, no differences were detected in bursa weight, however, a T x day interaction showed that T2 and T4 resulted in a larger BFA on d21. By d28 and 42, the BFA was larger for T3 (P=0.001). T x day interactions were also detected for cytokine modulations. Pentraxin levels were higher for T4 on d14 and T3 on d21 (P= 0.016). When compared to the control (T1), d0 IL-16 was higher for T4, by d14 it was reduced by all probiotic T, by d28 it was reduced by T3, and by d42 it was reduced by T3 and T4 (P=0.035). No differences in GABA expression were detected among T (P=0.12). However, a day effect demonstrated that GABA levels were lowest on d0, and increased as the bird aged (P=0.018). Contrary to the mice studies, the in ovo injection of Lactobacillus strains did not modulate GABA levels in broilers indicating no additional stress alleviation (welfare). However, the immuno-modulations obtained throughout the grow-out, with a single dose of probiotic, could be beneficial for broiler resistance to pathogenic challenges (heath). Further research should determine if these benefits are enhanced with the combination of in ovo and in feed delivery of Lactobacillus.
ID: 1084

DEVELOPMENT OF A MULTIPLEX PCR TO DETECT TRANSMISSIBLE ANTIBIOTIC RESISTANCE GENES IN ESCHERICHIA COLI

A.Taechavasonyoo*(1), R.Geier(2), E.Vang(3), J.Delago(3), W.Hirunpatrawong(4), T.Rehberger(3), A.Smith(3)

**Corresponding author** mail: Taechavasonyoo_apichaya@elanco.com (Apichaya Taechavasonyoo)

Sub-therapeutic levels of antibiotics to prevent disease and stimulate growth has been used in conventional poultry diets in Europe and the USA since the 1950's. The development of bacteria that are resistant to antibiotics used to treat human infections has resulted in regulatory changes limiting the use of antibiotics in livestock diets. Most broilers are therefore raised in production systems where antibiotics are not added to the diets. Reducing antibiotic resistance genes in the environment is one of the expected benefits of not adding growth-promoting antibiotics to poultry diets. The genomes of 111 Escherichia coli strains from broilers and turkeys collected in 2014 and 2015, before the concerted push to raise poultry without antibiotics, were analyzed for transmissible antibiotic resistance genes (ARGs). The average number of transmissible ARGs was six per isolate ranging from 0 to a maximum of 15 ARGs per strain. Seven ARGs from a diversity of antibiotic classes were selected to develop a multiplex PCR to monitor ARGs in avian E. coli. The genes selected were a dihydrofolate reductase (dfrA14) conferring resistance to trimethoprim, a tetracycline ePux pump (tetA), a beta-lactamase (blaTEM1), a macrolide 2'-phosphotransferase I (mphA) and dihydropteroate synthase (sulI) which enables folic acid biosynthesis in the presence of sulfonamide antibiotics. Two genes conferring resistance to aminoglycosides were selected an N-acetyltransferases, (aac3VIa) and an O-phosphotransferase (aph(3')-Ia). Specificity of the developed primers was determined by in silico PCR against the ResFinder database (v3.1.0 current as of 5/30/19). The multiplex PCR was used to examine poultry E. coli isolates from the USA and Thailand. A maximum of five of the seven genes were detected in any one isolate with the average number of genes being 1.7. The most prevalent ARGs in Thailand E. coli isolates were blaTEM1 and tetA at 70 and 60% respectively, while aac3Via and sulI were the most predominant ARGs in the USA at 43% and 42%. This multiplex assay will allow monitoring of transmissible antibiotic resistance genes in avian E. coli across countries and time.

<footnote>(1) Elanco (Thailand) Ltd., Bangkok, Thailand, (2) Arm & Hammer Animal and Food Production, Waukesha, United states, (3) Arm & Hammer Animal and Food Production, Waukesha, United states,</footnote>

<footnote>(4) Elanco (Thailand) Ltd., Bangkok, Thailand</footnote>

<footnote>*Presenting author</footnote>
ID: 1411

# NEWCASTLE DISEASE VIRUS INDUCES TESTICULAR DAMAGE AND DISRUPTS STEROIDOGENESIS IN SPECIFIC PATHOGEN FREE ROOSTERS

Z.Rehman*(1), S.Ren(2), S.Butt(3), A.Afzal(4), Y.Sun(2), C.Meng(2), C.Ding(2)

(1) Department of Poultry Science, Faculty of Veterinary and Animal Sciences, PMAS Arid Agriculture University, Rawalpindi, Pakistan, (2) Shanghai Veterinary Research Institute (SHVRI), Chinese Academy of Agricultural Sciences (CAAS), Shanghai, China, (3) Department of Pathology, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, Georgia, United States, (4) Institute of Statistics, Faculty of Economics and Management, Leibniz University Hannover, Hannover, Germany

*Presenting author

Corresponding author mail: zaib_rehman90@yahoo.com (Zaib Ur Rehman)

Newcastle disease (ND), which is caused by Newcastle disease virus (NDV), can cause heavy economic losses to the poultry industry worldwide. It is characterised by extensive pathologies of the digestive, respiratory, and nervous systems and can cause severe damage to the reproductive system of egg-laying hens. However, it is unknown whether NDV replicates in the male reproductive system of chickens and induces any pathologies. In this study, we selected a representative strain (i.e. ZJ1) of the most common genotype (i.e. VII) of NDV to investigate whether NDV can induce histological, hormonal, and inflammatory responses in the testes of specific pathogen free (SPF) roosters. NDV infection increased the expression of toll like receptor (TLR) 3, TLR7, MDA5, IFN-a, IFN-β, IFN-γ, IL-8, and CXCL12 in the testes of NDV-infected roosters at 5 days post-infection (DPI). NDV infection also decreased the plasma concentrations of testosterone and luteinizing hormone (LH) and the expression of their receptors in the testes. Severe histological changes, including decreases in the number of Sertoli cells and individualized, shrunken spermatogonia with pyknotic nuclei, were observed 3 DPI. At 5 DPI, the spermatogenic columns were disorganized, and there were fewer cells, which were replaced by necrotic cells, lipid vacuoles, and proteinaceous homogenous material. A severe decrease in the number of Sertoli cells and spermatids present in the lumen was also noted. A significant decrease in the mRNA expression of the steroidogenic acute regulatory protein, cytochrome P450 side-chain cleavage enzyme, and 3β-hydroxysteroid dehydrogenase in the NDV-infected group was observed relative to those in the control group (P < 0.05). Collectively, these results indicate that NDV infection induces a severe inflammatory response and histological changes, which decrease the steroidogenesis.

Key Words: roosters, Newcastle disease virus, steroidogenesis, cytokines, testis, steroidogenic acute regulatory protein, histopathology, 3β-hydroxysteroid dehydrogenase, cytochrome P450 side-chain cleavage enzyme
ID: 1412

NEWCASTLE DISEASE VIRUS INDUCED PATHOLOGIES SEVERELY
AFFECT THE EXOCRINE AND ENDOCRINE FUNCTIONS OF PANCREAS IN
CHICKENS

Z.Rehman*(1), S.Ren(2), A.Afzal(3), Y.Sun(2), C.Meng(2), C.Ding(2)

(1) Department of Poultry Science, Faculty of Veterinary and Animal Sciences, PMAS Arid Agriculture University, Rawalpindi, Pakistan, (2) Shanghai Veterinary Research Institute (SHVRI), Chinese Academy of Agricultural Sciences, Shanghai, China, (3) Institute of Statistics, Faculty of Economics and Management, Leibniz University Hannover, Hannover, China

*Presenting author

**Corresponding author mail:** zaib_rehman90@yahoo.com (Zaib Ur Rehman)

Newcastle disease virus (NDV) causes a highly contagious and
devastating disease in poultry, Newcastle disease (ND), which is
particularly characterized by extensive pathologies in the digestive,
respiratory and nervous systems. ND causes heavy economic losses to
the world poultry industry by decreasing growth rate and egg
productions, mortality, and morbidity. Although, significant advances
have been made in the vaccine development, but outbreaks are
reported in vaccinated birds leading to decrease growth rate. In this
study, we report the damage caused by the NDV infection in the
pancreas of vaccinated as well specific pathogen free chickens. The
histopathological examination of the pancreas showed sever damage
was reported in the form of partial depletion of zymogen granules,
acinar cell vacuolization, acinar cell necrosis and apoptosis, congestion
in the large and small vessels, sloughing of epithelial cells of pancreatic
duct, and mild perivascular edema. Increased plasma levels of
corticosterone, and somatostatin, were observed in NDV infected chicks
at 3 and 5-day post infection (DPI). Slight decrease in the plasma
concentrations of the insulin were noticed at 5 DPI. Significant changes
were not observed in the plasma levels of glucagon. Furthermore, NDV
infection has decreases the activity and mRNA expression of amylase,
lipase, and trypsin from the pancreas. Taken together, our findings
highlight that NDV induces extensive tissue damage in pancreas,
decrease the activity and expression of pancreatic enzymes and
increase plasma corticosterone and somatostatin.
ID: 1474

# ANTIMICROBIAL SUSCEPTIBILITY OF THREE DIFFERENT RIEMERELLA ANATIPESTIFER STRAINS IDENTIFIED BY MALTITOFS MS FROM A CLINICAL INFECTION OF BROILER CHICKENS

A.Tzora* (1), S.Skoufos(2), E.Bonos(2), K.Fotou(2), A.Karamoutsios(2), E.Barka(2), C.Voidarou(2), I.Skoufos(2)

(1) School of Agriculture, University of Ioannina, Arta, Greece, (2) Department of Agriculture, School of Agriculture, University of Ioannina, Arta, Greece

*Presenting author

Corresponding author mail: tzora@uoi.gr (Athina Tzora)

Riemerella anatipestifer (RA) is a contagious Gram-negative bacterium that is known to cause disease primarily in ducks and less frequently in geese and turkeys. In this case study, an infection of RA was identified in a broiler chicken rearing farm in Greece with a capacity of 70.000 broilers.

The clinical signs were observed at ages of 30 to 45 days with respiratory distress (dyspnea), white fluid feces and stunting. Morbidity was approximately 10% and mortality 5%. Post-mortem examinations displayed serositis, pericarditis, perihepatitis and airsacculitis. Oedematous swelling around the tibio-tarsal joints was observed in 50% of the examined birds. Tissue samples were collected from lesions and cultivated on Columbia sheep blood agar (CSB), containing 5% sheep blood, under microaerophilic conditions (5% CO2) at 37oC for 24 hours. Three bacterial isolates were identified by matrix-assisted laser desorption/isonisation time-of-flight mass spectrometer method (Microflex MALDI-TOF MS). Moreover, an antibiogram analysis was performed, using a pattern of 17 common antibiotics to advocate the most effective drugs for a targeted treatment. All the RA isolates were sensitive to ceftiofur, sulphamethoxazole-trimethoprim and amoxicillin, whereas all were resistant to gentamicin, tylosin, tetracyclin, colistin sulphate, spectinomycin, lyncomycin and oxytetracycline.

Acknowledgments: This research has been co-financed by Greece and the European Union (European Regional Development Fund) in context "Research – Create – Innovate" within the Operational Program (Competitiveness, Entrepreneurship and Innovation (EMANEK) of the NSRF 2014-2020. Project Code: TIE6K-03856. Acronym "GREEN POULTRY MEAT ANTIFREE".
Incubation physiology
ID: 627

STUDYING THE IMPACT OF LIGHT DURING INCUBATION AS A
POTENTIAL CANDIDATE TO INFLUENCE THE HATCHING TRAITS AND
SUBSEQUENT POST HATCH GROWTH PERFORMANCE OF BROILERS

J. Hussain*(1), M.Kashif yameen(1), A.Mahmud(1), S.Saima(3)

(1) University of Veterinary and Animal Sciences Lahore Pakistan, Lahore, Pakistan, (3) University of Veterinary & Animal Sciences Lahore Pakistan, Lahore, Pakistan

*Presenting author

Corresponding author mail: jibran.hussain@uvas.edu.pk (Jibran Hussain)

The chicken embryo starts responding to light from the second day of incubation. Just like temperature, moisture, turning, and ventilation, the light also has a significant role in embryonic growth. Providing light during incubation can help the embryo efficiently adjusting to its pre and post-natal environment. Yet, such a wonderful aspect remained neglected due to the possible interference of heat generated from light sources with incubator temperature. However, the advent of new light sources, especially, the LEDs (Light Emitting Diodes) have changed the dynamics of light emission, particularly no heat generation. Light from such sources can be utilized to modulate the embryonic growth as well as post hatch broiler performance. Keeping in view such a scenario, a study was planned to evaluate the effect of LED based light during incubation on hatching traits and post hatch performance of broilers. A total of 2250 eggs of three different commercial broiler strains were incubated under 0 (zero), 12 (twelve) and 24 (twenty four) hours lighting regimes to study the hatching traits in terms of egg moisture loss during incubation, early, mid and late embryonic mortality patterns, hatch window, hatch of fertile, and overall hatchability, post hatch stress response in terms of heterophil/lymphocyte ratio, physical asymmetry, feather score, and gait score as well as subsequent broiler growth performance in terms of feed intake, body weight gain, and FCR. A two-way factorial analysis was performed while significant means were separated through Duncan's Multiple Range test using SAS 9.1 software. Improved hatch window in terms of hatch synchronization, the higher hatch of fertile with reduced embryonic mortality each at early, mid and late stage with enhanced overall hatchability and superior chick quality were observed in response to 12 hours light stimulation during incubation. Likewise, a significant improvement was observed in stress response as well as the growth performance of broilers in response to light stimulation, especially for 12 hours stimulation. Considering the appreciable results of the present study, it is recommended to adopt 12 hours of light stimulation during incubation as a prerequisite factor. Development of antibacterial LEDs has opened new dimensions of research involving incubators: extensive research may help achieve improved hatching results, superior chick quality as well as decreased bacterial load, hence, improved broiler economics.
ID: 1111

EMBRYONIC CRITICAL PERIOD FOR SOMATOTROPHIC AXIS ELEVATION
BY IN-OVO GREEN LIGHT, AFFECTS BROILERS PERFORMANCE AT
MARKETING AGE

L.Dishon*(1), N.Avital-cohen(2), J.Bartman(2), S.Zaguri(2), I.Rozenboim(2)

(1) Faculty of Agriculture, Food and Environment Robert H.Smith, Rehovot, Israel, (2) The Robert H Smith Faculty of Agriculture, Food and Environment, the Hebrew University of Jerusalem, Rehovot, Israel

**Corresponding author mail:** dishonliron@gmail.com (Liron Dishon)

In ovo green light (GL) elevates somatotropic axis expression and broilers growth. The critical in ovo period for somatotropic axis stimulating by GL, and increase performance at marketing age was studied.

420 equal weight fertile broiler eggs (Cobb 500), divided into 7 equal treated groups: 1. Incubated at dark condition (Negative Control), 2. GD0-20: GL photostimulated between embryonic day ED0-ED20 (hatching day) (positive control), 3. GD15-20: GL photostimulated between ED15-20, 4. GD16: GL photostimulated on ED16, 5. GD17: GL photostimulated on ED17, 6. GD18: GL photostimulated on ED18 and 7. GD18-ED20: GL photostimulated between ED18-20. After hatching, the chicks were tagged according to the in ovo treatment group and were transferred to controlled rooms, where they were raised under standard conditions. The chicks were weighed weekly and on day 35 blood samples were taken from 10 chicks of each treatment group. Afterward, each one was weighted and sacrificed for tissue samples (such as hypothalamus, pituitary, liver, and breast muscle) following weighting breast muscle, liver, and abdominal fat weights. The blood samples were used for GH ELISA assay. Hypothalamus, pituitary, liver, and breast muscle samples were collected for mRNA gene expression of GHRH, GH, GHR (Growth Hormone Receptor) and IGF-1 (Insulin-Like Growth Factor 1), by Real-Time PCR. After finding no significant interaction between treatment and age, all statistical analyses were conducted with the JMP software using one-way ANOVA.

All treatment groups elevated body weight at day 35, between 40g and 180g, compared to the dark group. All treatment groups elevated plasma GH levels, compared to the dark group. All treatment groups, aside from GD17 and GD18, elevated breast muscle weight by 1-2% of body weight. GD0-20, GD15-20, and GD18-20 elevated liver GHR and IGF-1 gene expression on day 35.

In ovo green light photostimulation of broiler embryos, from ED18-ED20 significantly elevated growth and somatotropic axis activity similar to the positive control group. We suggest that the critical period for GL photostimulation acceleration of the somatotropic axis is between ED18-ED20.

<footnote>*Presenting author</footnote>
Ingestion and digestion
ID: 950

PROTECTED SODIUM BUTYRATE IN BROILER PRE-STARTER AND
STARTER DIETS IMPROVES THE PERFORMANCE AT SLAUGHTER AGE

M.Ferreira pires(1), F.Carvalho*(1), R.Almeida noleto(1), D.Pereira carvalho(1), S.Ferreira pires(1), D.Vanni jacob(2), J.Stringhini(1), M.Barcellos café(1), N.Mogyca leandro(1)

(1) Federal University of Goiás, Goiânia, Brazil, (2) Adisseo Brazil, Goiânia, Brazil

*Presenting author

Corresponding author mail: fabyolab@hotmail.com (Fabyola Carvalho)

Recognizing that butyrate plays an important role in maintaining the
intestinal mucosa, increase the length of villi, improvement nutrient
metabolism coeficients and as an antimicrobial agent, it is expected
that with its use, there will be an improvement in nutrients absorption
and consequently improved performance of broilers. Thus, the objective
of this study is to evaluate the performance of fed broilers with pre-initial
and initial rations and with different levels of sodium butyrate protected
and to verify its effect until the final rearing period. The experiment was
carried out in experimental facilities from the Federal University of Goiás.
784 day-old Cobb 500® linhage chicks with an average weight of 46g
were purchased. The design was installed in dellienament casually,
four trataments, with seven repetitions with 28 birds. Thus, four levels
of protected sodium butyrate in the pre-initial and initial phase (0; 150;
225 and 300 g / t) were studied. It was used as a source of protected
butyrate the commercial additive Adimix®Precision, Nutriad (30%
sodium butyrate). The rations were formulated according to the
requirements defined by the integrating company. The diets contained
anticoccidial agent and did not contain antibiotics. On the first, seventh,
21st, 35th and 42nd days of age, poultry, feed and leftover weighing
were performed to calculate zootechnical performance data (final
weight, weight gain, feed intake, corrected alimentar conversion and
viability). Data were submitted to polynomial regression analysis, with
the aid of the R (2016) statistical program. Results presented from 1 to
42 days show the beneficial effect of protected sodium butyrate on the
feed rations broilers, where increasing linear effects for final weight,
weight gain and viability were observed.

In conclusion, the use of protected sodium butyrate either in the pre-
initial or the initial ration improved broiler performance until the final
rearing phase, with the recommended level of 300 g / t.
THE EFFECT OF EXOGENOUS ALPHA-AMYLASE IN DIET WITH PEA AND FABA BEAN ON BROILER CHICKENS PERFORMANCE AND NUTRIENT UTILIZATION

M.Hejdysz*(1), K.Perz*(2), S.Kaczmarek(2), A.Rutkowski(2)

(1) Poznan University of Life Sciences, Poznan, Poland, (2) University of Life Sciences in Poznan, Poznan, Poland

*Presenting author

Corresponding author mail: marcin.hejdysz@up.poznan.pl (Marcin Hejdysz)

Pea and faba bean seeds belong to legumes commonly grown around the world. However, these legume also contains variable amounts of anti-nutritional factors and high content of resistant starch (from 15 to 20% of dry matter), which can negatively affect nutrient utilization and animal performance. One process which can improve the nutritional value of pea and faba bean seeds is the use of exogenous enzymes, through the reduction of resistant starch content. Therefore, this study tries to determine the effects of exogenous enzymes in diet with faba bean and pea on broiler chickens performance and nutrient utilization.

The experiment, which lasted 35 days, was carried out on 960 male one-day old ROSS 308 chickens. The birds were randomly divided into 4 experimental groups. Each group consisted of 30 replications (8 birds in each replication). Feed and water were given ad libitum. The diets were isocaloric and isonitrogen. The first and third groups, diet contained 30% of faba bean and pea respectively. In the second and fourth groups, were used the same legumes components but with exogenous alpha-amylase with amount 0.014 %. The trial was divided for starter (0-14 day) and grower (15-35 day) period. At the end of the experiment, weight gains, feed intake and feed conversion ratio were calculated. On day 35 of the experiment, all chickens were sacrificed by cervical dislocation and digesta were flushed from the terminal ileum and pooled (six birds/sample) to provide enough material for chemical analyses (n = 10). The obtained results were subjected to a two-way analysis of variance (ANOVA), and then in order to determine the significance of differences between groups' means, the Duncan's range test was used, with the significance level of P < 0.05.

There was not confirmed interaction between experimental factors for each growth performance parameters. The use of enzyme in legume diets statistically significant improve body weight gains of broilers. Feed conversion ratio was statistically significant lower after enzyme supplementation. Nutrients utilization (e.g. starch, crude protein) were improved after enzyme utilization. However AMEN value did not different statistical significant between groups with and without exogenous alpha-amylase.

As a conclusion, the use of exogenous alpha-amylase improve broiler chicken performance.
Meat processing
ID: 1460

# COLIBACILLOSIS IN BROILER CHICKENS: WHAT IS THE IMPACT ON CARCASS QUALITY? A FRENCH RETROSPECTIVE STUDY

C.Bera*(1), K.Adjou(2), N.Galliard(3), P.Arne(2), C.Chimienti(1)

(1) VET&SPHERE Réseau Cristal, Quintin, France, (2) Ecole Nationale Vétérinaire d'Alfort, Maisons-alfort, France, (3) ZOETIS, Malakoff, France

Corresponding author mail: claire.bera.vet@gmail.com (Claire Bera)

Colibacillosis is the main bacterial disease in the broiler chickens industry worldwide. This is the first reason of condemnation at slaughtering in Europe, resulting in massive economic losses in the poultry industry. However, only a few data are available in France to date.

The goal of this study was to describe and to quantify the colibacillosis impact on carcass quality at slaughtering, by analysing retrospectively slaughtering data and food chain information from July 2017 to June 2018 in four slaughterhouses in Britain (France). The flocks were classified into three groups: "early colibacillosis" meant in the first ten days of rearing, "late colibacillosis" meant in the last fifteen days of rearing and "flocks with no colibacillosis history". Slaughtering data were averaged for multiple slaughter days for male and female separately. The presence of other diseases and related treatments were taken into account. The statistical analysis was based on linear mixed models. Least squares means of slaughtering data from the corresponding model were compared between the three groups, using the Kenward-Roger degrees of freedom. The significant level of p-value was defined at 0.1.

Among 9713 collected flocks, the colibacillosis annual prevalence in standard broiler chickens in Britain was about 22 % in females, two-thirds of which were early forms, and was about 25 % in males, two-thirds of which were late forms.

Carcass yield in males and fillet yield in females were lower in case of early colibacillosis, compared to flocks free of colibacillosis (-0.276 %, p=0.06 and -2.503 %, p<0.01 respectively). Moreover, live mean weight was higher in male flocks exposed to colibacillosis (+78 g, p<0.01 in case of early colibacillosis and +26 g, p=0.06 in case of late colibacillosis). This offsets partial condemnations losses in case of late colibacillosis (-0.029 %, p=0.06), unlike total condemnations losses which were higher (farm/slaughterhouse) (+0.204 %, p<0.01). Mortality during transport and flock heterogeneity were not significantly different between the three groups.

Colibacillosis significantly affect live mean weight in males, carcass and fillet yields in females, which are the main criteria involved in the major economic and social issues today.

<footnote>*Presenting author</footnote>
Metabolism
# SULFATION AS A PATHWAY OF METABOLISATION OF DEOXYNIVALENOL TO DEOXYNIVALENOL-SULPHATE IN CHICKENS

I.Riahi*(1), A.Pérez(2), V.Marquis(3), A.Ramos(4)

(1) Institute for Food and Agricultural Research and Technology (IRTA), Animal Nutrition, Constanti, Spain, Tarragone, Spain, (2) IRTA, Animal Nutrition, Tarragone, Spain, (3) Phileo by Lesaffre Animal Care, Toulouse, France, (4) Food Technology Department, University of Lleida., Lleida, Spain

*Presenting author

**Corresponding author mail:** insaf.riahi@irta.cat (Insaf Riahi)

Recent experiments observed that deoxynivalenol (DON) is extensively metabolized to DON-sulphate (DON-S) in chickens. The aim of the study was to evaluate the most relevant biomarker in broiler chickens plasma of exposure of DON after intravenous and oral administration.

A toxicokinetic study was performed with twenty- four 35 day-old chickens Ross 308. Doses of 0.75 and 2.25 mg DON/kg of BW were administrated intravenously and orally to the chickens. The administrated dose of 0.75 mg/kg BW corresponds to 5 mg DON/ kg of poultry feed (maximum level recommended by the European Commission). Blood was collected at different time points at (0) pre-administration and at 15, 35, 60, 75 and 120 minutes post administration. Samples were centrifuged for 10 min at 2851*g* and plasma was stored at -20°C until further analysis. Plasma samples were prepared by adding acetonitrile to precipitate proteins, pursued by evaporation of the supernatant.

Chromatographic analysis by LC-MS/MS revealed that DON-S was the major metabolite of DON in broiler chickens blood after intravenous and oral administration.

The maximal concentration of DON in plasma was 36.5 and 101.5 ng/ml after IV administration of 0.75 and 2.25 mg of DON /kg BW, respectively. After oral administration of the same levels of the mycotoxin to broiler chickens, DON concentration in plasma was below the LOQ of 5 ng/m, and only DON-S was identified. Since no commercial standard of DON-S is available, the results were evaluated using the absolute chromatographic peak areas. The maximum areas of DON-S were 2,178,419 and 7,053,108 for the intravenous administration at low and high level, respectively and were 122,623 and 713, 515 for the oral bolus administration at low and high level, respectively. The peaks of this metabolite in plasma were achieved after 15 min post administration.

The mean area under the curve (AUC0-2h) for IV application of DON at low level was 28.48 and at high level was 76.81counts. The mean area under the curve (AUC0-2h) of DON-S for IV application at low level was 952,087 counts and at high level was 2,824,109 counts. The mean area under the curve (AUC0-2h) of DON-S for oral application at low level was 52,518 counts and at high level was 343, 970.

This study confirms the results recently reported, that DON-S is the most suitable biomarker for DON exposure in broiler chickens.

**Key words:** deoxynivalenol, deoxynivalenol-sulfate, biomarker, peak areas
ID: 988

PERFORMANCE, NUTRIENTS METABOLIZABILITY AND INTESTINAL HISTOMORPHOMETRY OF BROILERS SUPPLEMENTED WITH CHONDROITIN AND GLUCOSAMINE SULFATES ON RATION

M.Cafe*(1), J.Martins(2), S.Sgavioli(2), G.Carvalho(2), M.Costa(2), J.Stringhini(2)

(1) Universidade J. e Federal de Goias, Goiania - goias, Brazil, (2) Universidade Federal de Goias, Goiania, goias, Brazil

*Presenting author

Corresponding author mail: mcafe@ufg.br (Marcos Cafe)

The objective was to evaluate performance, nutrient metabolizability and intestinal histomorphometry of broilers supplemented with chondroitin and glucosamine sulfates on ration. The experiment was conducted in a completely randomized design, in a 2 x 2 factorial scheme (two levels of chondroitin sulfate: 0 and 0.10%, and two levels of glucosamine sulfate: 0 and 0.30%), with eight replicates of 15 birds. Performance was evaluated at seven and 21 days old, feed nutrient metabolization performed from 18 to 21 days old, and small intestine histomorphometry at 21 days old. Data were submitted to variance analysis and means were compared by t test at 5% probability. Pearson correlation analysis was performed between the variables of nutrient metabolizability and intestinal histomorphometry using the t test 5% probability. Final weight and weight gain were influenced by glucosamine sulfate at 21 days old, with higher values for the chickens that received supplementation. There was interaction of sulfates for nitrogen metabolic coefficients, mineral matter and calcium. Inclusion of 0.10% of chondroitin sulfate in the sulfate-free diet of glucosamine presented a lower coefficient of nitrogen metabolism. Chondroitin sulfate or glucosamine sulfate ratios had higher mineral matter metabolic coefficients and calcium. Broilers supplemented with glucosamine sulfate, without chondroitin sulfate and 0.10% chondroitin sulfate, presented higher villus height than birds that were not supplemented glucosamine sulfate. There were strong positive correlations between dry matter metabolizable coefficient and ratio villus:crypt duodenum (r = 0.93) and jejunum (r = 0.88). Between nitrogen metabolizable coefficient and apparent metabolizable energy corrected by nitrogen balance (r = 0.99), and the ratio villus:crypt duodenum (r = 0.87) and jejunum (r = 0.97). Between mineral metabolizability coefficient and villus height and jejunum crypt depth (r = 0.96 and r = 0.94, respectively) and ileum (r = 0.96 and r = 0.98, respectively).

It was concluded that supplementation with glucosamine sulfate in broiler ration increased weight gain, and stimulated instestinal histomorphometry. Sulfates alone promoted greater metabolism of minerals.

Key words: digestibility, intestinal health, polysulfated glycosaminoglycans.
ID: 1104

# TREATMENT OF CHICKEN MACROPHAGES WITH BUTYRATE INCREASES GLUCOSE METABOLISM IN VITRO.

F.Perry*(1), L.Aeschleman(1), R.Arsenault(1)

(1) University of Delaware, Newark, United states

*Presenting author

**Corresponding author mail:** famper@udel.edu (Famatta Perry)

Butyrate and other short chain fatty acids (SCFA) are produced by many bacteria in the gut and these SCFA may influence energy metabolism of a variety of cells in the gut. Studies have shown that these SCFA also increase fatty acid oxidation which aids in the regulation of inflammation and maintenance of barrier integrity. Past research has provided evidence that the incorporation of these SCFA in bird feed can help boost immune responses to fight off pathogens. The objective of this project was to examine the impact of butyrate on the energy metabolism of macrophages in vitro. Using the seahorse XFp analyzer, a metabolic flux assay measuring the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) was performed on chicken-macrophage like (HD-11) cells infected with Salmonella with and without pretreatment with 4mM butyrate compared to non-infected controls. Fifty thousand HD-11 cells were infected with Salmonella Enteritidis (SE) or Salmonella Heidelberg (SH) at an MOI of 1:100 and their metabolic activities were measured over 6 hours. The measurements reported as pmol/min for OCR and mpH/min for ECAR was compared using a Student's T-Test for each group. The results showed no difference in OCR of uninfected cells treated with butyrate compared to uninfected cells without butyrate treatment. However, there were significant differences shown in the ECAR between uninfected cells treated with and without butyrate overtime. There was a 30%-40% increase in the ECAR which measures glycolysis (a sign of inflammation) of butyrate treated controls compared to the untreated controls. Metabolic flux results of SE and SH infected cells post-butyrate treatment showed an increase in ECAR and OCR of uninfected cells compared to infected cells. Further analysis showed a difference in trend between infected cells treated with butyrate compared to their untreated counterparts. SH infected cells at 1.5 hours post-butyrate treatment showed a significant increase of 34.22 mpH/min in ECAR compared to 27.34 mpH/min SH infection alone. This increase was maintained for 1 hour (p-value < 0.01). Using the seahorse metabolic flux assay, we demonstrated the metabolic impact of butyrate on macrophages in vitro. Here we show that, unlike in epithelial cells, butyrate in macrophages may increase inflammation, as measured by an increase in ECAR. We also revealed that treatment of SH infected cells with butyrate leads to an increase in glucose metabolism by the cells.
ID: 1319

THERMALLY OXIDIZED VEGETABLE OIL INFLUENCES GROWTH PERFORMANCE, CARCASS CHARACTERISTICS, GUT HEALTH, NUTRIENTS DIGESTIBILITY AND SERUM BIO-CHEMISTRY IN BROILERS

S.Qaisrani*(1), G.Yaseen(1), M.Sarfraz(2), F.Bibi(3), S.Naveed(2), T.Pasha(4)

(1) University of Veterinary and Animal Sciences, Lahore, Lahore, Pakistan, (2) University of Veterinary and Animal Sciences, Lahore, Pakistan, (3) Bahauddin Zakriya University, Multan, Pakistan, (4) University of Education, Lahore, Pakistan

*Presenting author

Corresponding author mail: shafqat.qaisrani@uvas.edu.pk (Shafqat Nawaz Qaisrani)

The objectives of this study were to evaluate the effects of various dietary levels of thermally oxidized vegetable oil on growth performance, carcass characteristics, gut morphology, nutrients digestibility and serum bio-chemistry in broilers. Ross 308 straight-run broilers (n= 192, d 0) were assigned to 4 dietary treatments with 6 replicate pens in a completely randomized design. The treatments included; 1) fresh (1 mEq/kg) vegetable oil (FVO); 2) low oxidized (20 mEq/kg) vegetable oil (LVO); 3) moderately oxidized (40 mEq/kg) vegetable oil (MVO); and 4) highly oxidized (60 mEq/kg) vegetable oil (HVO) with 5% dietary inclusion. The data analysis with SAS (9.1) indicated that feed intake (P = 0.034) and body weight gain (P = 0.041) were decreased leading to an inferior feed conversion ratio (P = 0.024), over the entire (0 to 5 wk) experimental period, in broilers consuming diets containing MVO and HVO compared with those fed LVO and FVO. Carcass weight and dressing percentages decreased (P = 0.001), whereas breast and leg quarter yields and giblets weight remained unaffected (P = 0.420) by the level of oxidation. The duodenal villus height (VH) was decreased by 1.6, 24 and 28%, and crypt depth (CD) increased by 4, 8, and 14%, whereas villus height to crypt depth ratio (VCR) was decreased by 7, 18 and 17% in broilers fed diets containing LVO, MVO and HVO, respectively, compared with those consuming diets with FVO. There was a decrease (P< 0.05) in the ileal digestibility of protein, amino acids and crude fat, in the broilers consuming diets containing LVO, MVO and HVO compared with those fed diets with FVO. The serum cholesterol level, similarly, was decreased by 17, 21 and 23% in broilers consuming LVO, MVO and HVO, respectively, compared with those fed diets with FVO. In conclusion, dietary inclusion of thermally oxidized oil with oxidation level > 20 mEq/Kg may lead to an inferior gut health and nutrients digestibility resulting in a poorer growth performance. Inclusion of thermally oxidized oil, therefore, with oxidation level < 20 mEq/Kg may result in more sustainable and economical poultry production by replacing the costly fresh vegetable oil in poultry diets.

Key words: oxidized vegetable oil, growth performance, gut health, nutrient digestibility, amino acids, broilers.
# Mineral nutrition
ID: 207

# HYDROXYCHLORIDE ZINC, COPPER, MANGANESE USED IN LAYING HENS' DIET AFFECTS TIBIA TRAITS AND EGG MINERAL DEPOSITION

T.Nguyen*(1), R.Swick(1), M.Toghyani(1), N.Morgan(1), J.Roberts(1)

(1) School of Environmental and Rural Science, University of New England, Armidale, NSW 2351, Australia

*Presenting author

Corresponding author mail: tnguye85@myune.edu.au (Thi Thanh Hoai (Anna) Nguyen)

Hydroxychloride trace minerals (HTM) have covalent bonds and thus are less reactive both in the feed and in the digestive tract compared to other inorganic trace minerals, which contain ionic bonds (ITM) such as sulphate forms. Therefore, HTM may have higher bioavailability than ITM. This study aimed to evaluate the effects of replacing sulphate forms of zinc (ZnSO4), manganese (MnSO4) and copper (CuSO4) (ITM) with HTM sources on tibia bone traits and mineral content in eggs of layers during post-peak production. Hy-line Brown hens at 45 wk old (n = 600) were randomly distributed into 300 cages, with 10 cages (20 birds) per replicate and 15 replicates per treatment. There were two treatments (ITM or HTM) with Zn, Mn and Cu supplemented at 80 ppm, 80 ppm, and 15 ppm, respectively, for 12 weeks. At the end of weeks 2, 6, and 12 of the study, two eggs per replicate were collected to determine mineral content in the eggshell and yolk. At the end of the study, tibia bones were excised from two representative birds per replicate after euthanasia to measure breaking strength and mineral content. The tibias were subjected to breaking strength by an Instron instrument. The content of Ca, Zn, Cu and Mn in the eggs and tibias were measured using inductively coupled plasma optical emission spectroscopy. Tibia data were analysed via a one-way ANOVA and mineral deposition in the eggs was analyzed via two-way ANOVA including 2 diets and 3 ages. Means with significant differences were separated by Tukey's HSD test at P < 0.05. The tibia samples from birds fed HTM contained more Cu (1.36 µg/g vs. 1.16 µg/g, P < 0.05) and tended (P = 0.075) to have higher breaking strength than those fed ITM. No significant interaction between diet and age for mineral deposition in the eggshell and yolk was observed (P > 0.05). Birds fed the HTM diet had increased Cu concentration in the eggshell compared to those fed the ITM (1.084 µg/g vs. 0.936 µg/g, P < 0.01). As the birds aged, the Ca and Cu content of the shell increased (P < 0.01). The HTM group had higher Zn content in the yolk than the ITM group (84.4 µg/g vs 82.6 µg/g, P < 0.05), but other minerals were not affected by mineral source. The yolk Mn content was lower at week 2 than at week 6 (3.24 µg/g vs. 3.53 µg/g, P < 0.05). The results indicate that replacing sulphate sources of Zn, Mn and Cu with hydroxychloride sources resulted in a tendency for increased tibia breaking strength and increased mineral accumulation in eggs.
ID: 274

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

*Presenting author

**Corresponding author mail:** bolufemi@purdue.edu (Olufemi Babatunde)

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 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 < 0.05) body weight (BW), weight gain (WG), feed intake (FI) and a higher (P < 0.01) feed conversion ratio (FCR) as compared with birds fed the PC. With increasing PP concentrations, there was a linear decrease (P < 0.01) in WG by 5% in phase 1 and 6% in phase 2. While FI was decreased (P < 0.01) with increasing PP, there was no effect on FCR in phase 1. However, in phase 2, increasing PP had no effects on FI but increased (P < 0.01) FCR in broiler chickens. Increasing phytase levels linearly improved (P < 0.01) BW of broiler chickens by a range of 9 to 18% in phase 1 and 9 to 14% in phase 2. Adding 4,000 FTU/kg improved (P < 0.01) WG by 22 and 20% in phase 1 and 2, respectively. Similarly, FI was linearly increased (P < 0.01) in both phases by approximately 14 and 11% respectively with increasing phytase levels. There was a linear decrease (P < 0.01) in FCR and a quadratic decrease (P < 0.01) in FCR of birds fed phytase-supplemented NC diets as compared with birds fed the NC with no phytase inclusion in phase 1 and 2 respectively. In conclusion, increasing PP concentrations had negative effects on the growth performance of broiler chickens. However, phytase supplementation alleviated the negative impacts of PP by increasing P availability for use in the various biological processes required for growth in broiler chickens.
ID: 381

EFFECTS OF DIFFERENT SOURCES AND LEVELS OF COPPER AND MANGANESE ON PERFORMANCE AND MEAT QUALITY OF BROILER CHICKENS

K.Von Zuben Augusto*(1), P.Michelin Groff-Urayama(2), Y.Han(1), C.Yutto Oura(2), T.Souza dos Santos(2), J.Moraes Cruvinel(2), F.Kaiser de lima-krenchinski(2), J.Santos Batistioli(2), C.Toledo Santos(2), J.Sartori(2)

(1) Trouw Nutrition R&D, Ameersfort, Netherlands, (2) São Paulo State University (UNESP), Botucatu, Brazil

*Presenting author

Corresponding author mail: karolina.augusto@trouwnutrition.com (Karolina Von Zuben Augusto)

The most recent sources of copper (Cu) and manganese (Mn) in broiler nutrition are hydroxychlorides (Intellibond® C - IBC and Intellibond® M - IBM, respectively), which are known for their great strength in chemical bonding more stable in the diet and in gastrointestinal tract. Thus, the objective of the present study was to evaluate the effect of different sources and levels of Cu (Cu Sulfate monohydrate - CSM and IBC) and Mn (Mn sulfate monohydrate - MSM and IBM) on performance (1 to 28d-old), and meat quality and breast yield (42d-old). A total of 1,920 1-d-old male Cobb chicks were distributed in a completely randomized design with a factorial arrangement of 2x3+2: IBC (15 and 150 ppm) x IBM (40, 80, 120 ppm) + 15 ppm CSM with 80 ppm MSM, and 150 ppm CSM with 120 ppm MSM, with 8 treatments and 8 replicates of 30 birds each. The diets were based on corn and soybean meal according to the recommendations of each phase: pre-starter (d1-d7), starter (d8-d21), grower (d22-d35), and finisher (d36-d42). At the end of the experiment, three birds per replicate were slaughtered for breast yield assessment, and two birds for breast meat quality analyses: water-holding capacity, cook loss, shear force, pH, and color. Data were analyzed with Minitab® 18, and the means were compared by Tukey and Dunnett tests (P<0.05). Broilers fed diets containing 150 ppm IBC were better than those fed diets containing 15 ppm for feed conversion ratio - FCR (P=0.009) and feed intake (P=0.001). When analyzing only the sulfate sources, the broilers supplemented with 150 ppm CSM with 120 ppm MSM had better weight gain and FCR than those supplemented with 15 ppm CSM plus 80 ppm MSM. For meat quality it was observed that the inclusion of 150 ppm IBC increase lightness (L*) (P=0.001) and yellowness (b*) (P=0.030) when compared to 15 ppm, whereas 40 ppm IBM increases lightness (P=0.036) when compared to 120 ppm IBM. For broilers receiving 150 ppm IBC, the breast yield was increased (P=0.030) by the inclusion of 40 ppm IBM, when compared to broilers receiving 120 ppm. In conclusion, the supplementation of broiler diets with 150 ppm IBC improves FCR at 28 d-old. Furthermore, by supplying 150 ppm IBC with 40 or 80 ppm IBM, enhance lightness, yellowness and breast yield of broiler chickens at 42 d-old.
DIETARY SUPPLEMENTATION WITH SODIUM BICARBONATE OR SODIUM SULFATE AFFECTS EGGSHELL QUALITY BY ALTERING EGGSHELL ULTRASTRUCTURE AND COMPONENTS INLAYING HENS

Y.Fu(1), S.Wu(1), H.Zhang(1), J.Zhou(1), G.Qi(1), J.Wang*(1)

(1) Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China

**Corresponding author mail:** qiguanghai@caas.cn (Guang-Hai Qi)

This study compared the effects of dietary Na supplementation in sodium bicarbonate (NaHCO3) or sodium sulfate (Na2SO4) forms on laying performance, eggshell quality, ultrastructures and components in laying hens. A total of 576 29-wk-old Hy-Line Brown laying hens were randomly allocated to 8 dietary treatments that were fed a Na-deficient basal diet (0.07% Na, 0.15% Cl) supplemented with Na2SO4 or NaHCO3 at 0.08, 0.18, 0.23, or 0.33% Na for 12 wks. We examined the eggshell mechanical quality every 4 wks, and further analyzed eggshell ultrastructures and components at the end of the trial. All analyses were performed using SPSS 19.0 for windows (SPSS Inc., Chicago, IL). The data were analyzed using one-way ANOVA and regression analysis, respectively. The statistical test of comparisons between the Na2SO4 and NaHCO3 forms were further assessed using covariance analysis. Dietary Na supplementation resulted in quadratic increases of eggshell breaking strength in both Na2SO4 and NaHCO3 added groups (P < 0.05), and Na2SO4-fed groups had a quadratic increase in the eggshell ratio at wk 12 (P < 0.05). Compared with supplementing 0.08% Na, dietary supplementation of 0.23% Na increased the effective thickness (P < 0.05) in both Na2SO4 and NaHCO3 added groups, while decreasing the thickness and knob width of mammillary layer (P < 0.05). A linear increase on the calcium content of shell was only observed with Na supplementation from NaHCO3 (P < 0.05). Dietary Na addition had a quadratic increase on uronic acid contents of membrane in NaHCO3-fed groups (P < 0.05). The sulfated glycosaminoglycans (GAGs) contents of membrane increased linearly with dietary Na supplementation (Na2SO4, P = 0.035; NaHCO3, P = 0.037). Additionally, compared with NaHCO3-fed groups, Na2SO4-fed groups had higher eggshell breaking strength, thickness, eggshell weight ratio, effective thickness and the sulfated GAGs content of calcified eggshell.

Overall, dietary supplementation of NaHCO3 or Na2SO4 could increase eggshell breaking strength, which may be related to increased sulfated GAGs contents in eggshell membranes and improved ultrastructures. Higher eggshell strength, thickness and eggshell ratio could be obtained when the diet was supplemented with 0.23% Na from Na2SO4.

<footnote>*Presenting author</footnote>
ID: 887

EFFICACY OF HIGH DOSES OF PHYTASE ON PERFORMANCE, TIBIA MINERALIZATION, APPARENT ILEAL NUTRIENT DIGESTIBILITY, INTESTINAL MORPHOLOGY AND GALT ASSOCIATED MEASUREMENTS IN BROILER CHICKENS

J.Zentek*(1), B.Martinez-vallespin(1), P.Ader(2), K.Männer(1)

(1) Institute of Animal Nutrition, Berlin, Germany, (2) BASF SE, Lampertheim, Germany

*Presenting author

**Corresponding author mail:** juergen.zentek@fu-berlin.de (Jürgen Zentek)

Phytic acid has a negative effect on phosphorus and nutrient digestibility and might affect the intestinal mucosa. The aim of the experiment was to investigate the response to different levels of phytase supplemented to a basal diet rich in phytic acid.

From 1 to 21 days of age, broiler chickens were assigned to five dietary treatments (T1-T5): A "phytate-free" diet based on Hermetia meal and corn starch (T1), and phytate-rich diets with corn, soybean meal and sunflower meal (T2-T5). The phytate-rich diets were prepared without (T2) and with supplementation of Natuphos® E (BASF SE, Germany) at levels of 500, 1,000 and 3,000 FTU/kg (T3-T5). For each treatment, 14 replicates with 2 broilers/cage (Q) were used. Performance was measured on a weekly basis, the apparent ileal digestibility of nutrients was measured using titanium oxide. The tibia was analyzed for minerals, histo-morphometry and immune histochemistry were performed in the jejunum and caecum. Statistical methods included ANOVA and Tukey test ($P < 0.05$).

Broiler chickens fed the T1 diet reached a lower body weight compared to all other groups. Increasing phytase inclusion to the phytate-containing basal diet resulted in improvements of performance, apparent ileal digestibility of crude protein ($p < 0.001$) and amino acids up to dosage of 1,000 FTU/kg feed. The highest relative apparent ileal phosphorus digestibility was observed with the phytate free diet (65.5 %), phytase supplementation resulted in dose-dependent positive responses of relative apparent ileal digestibility of phosphorus ($p < 0.001$). Birds fed the diet T2 had reduced tibia ash and phosphorus compared to T1. Increasing phytase resulted in a dose dependent improvement of mineralisation ($p < 0.001$). The results of histomorphological measurements in the jejunum and in the caecum showed higher villus lengths and an impact on crypt depth when comparing T1 with the other treatments but also T2 with phytase supplemented groups. T1 had a significantly lower number of intraepithelial lymphocytes than T2 ($p < 0.001$). The supplementation with phytase reduced the numbers of intraepithelial lymphocytes compared to T2 ($p < 0.001$). In T5 the number was numerically the same as in T1.

In conclusion, an increase in the phytase dosage led to a significant improvement in the measured parameters compared to the standard recommendation. The observed impact of phytase on gut mucosa requires further validation.
ID: 1092

EFFECT OF DIETARY SUPPLEMENTATION OF ORGANIC AND INORGANIC SE ON PERFORMANCE AND ANTIOXIDANT RESPONSE IN COMMERCIAL BROILER CHICKENS

B.Prakash*(1), M.Prasad(2), S.Rama rao(3), M.Raju(3), S.Paul(3), A.Kannan(3), C.Reddy(3)

(1) ICAR-Directorate of Poultry Research, Hyderabad, India, (2) Animal Nutrition Department, P.V.N.R Telangana Veterinary University, Hyderabad, India, (3) ICAR-DPR, Hyderabad, India

*Presenting author

**Corresponding author mail:** drbhukyaprakash@gmail.com (Bhukya Prakash)

Selenium is an important for optimum immune response and considered as a functional part of the antioxidant properties through selenoproteins in chickens. The organic form of Se is rapidly absorbed through an active transport. In contrast, the inorganic Se absorbed slowly by simple diffusion. Therefore, the study was carried out to determine the effects of supplementing organic and inorganic Se to record the performance, immune and anti-oxidant response in broiler chickens. One-day-old Vencobb-400 (50.1±0.84 g; n=280) broiler chicks were distributed randomly into 7 treatments having 8 replicates with each 5 chicks and housed in deep litter (floor pens 6 × 6 ft.) for a period of 42 days. The basal diet (BD) was formulated without Se supplementation (Diet -I), BD plus 0.15, 0.30 and 0.45 mg/kg organic Se (Diet II, III and IV, respectively) and BD plus 0.15, 0.30 and 0.45 mg/kg inorganic Se (Diet V, VI and VII, respectively). Determined feed intake (FI), body weight gain (BWG), immune parameters and mRNA expression profile of hepatic selenoproteins genes. The data was analysed using the general linear model procedure. The model included different dietary treatments as source of variation. Treatment means were compared using Tukey's test. The BWG, FI, feed conversion ratio (FCR) and slaughter parameters did not differ (P>0.05) among various dietary treatments. The lipid peroxidation (LP) and activity of RBC catalase were lower (P<0.05) in groups fed diet organic Se compared to those groups fed BD. The activity of GPx was higher among the groups fed diet supplemented organic Se compared to those groups fed BD. Supplementing diets with organic or inorganic Se did not affect LP, RBC catalase, GPx, humoral immune and cell mediated immune response in the present experiment. The expression levels of SepW was higher (P<0.01) among the groups fed diet supplemented 0.3 mg/kg organic Se. Expression level of GSH-Px1, TrxR and GSHPx3 were higher (P<0.01) among the groups fed diet supplemented 0.45 mg/kg organic Se compared to those groups fed diet supplemented inorganic Se or BD.

Therefore, it has been concluded that supplementing diet with organic or inorganic Se did not affect performance, immune parameters and activities of anti-oxidant enzymes. However, supplementing organic Se improved hepatic expression of selenoproteins genes in the present study.
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(2), 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, (2) Faisal Group of Companies, Multan, Pakistan, Na, Pakistan

*Presenting author

Corresponding author mail: DrShahzadBalouch@Yahoo.Com (Shahzad Ashraf)

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 (SY), 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. Glutathione peroxidase activity, total antioxidant capacity and antibodies titer against Newcastle disease virus was also measured. 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
Molecular genetics
ID: 110

# IDENTIFICATION OF INDEL(S) AND SNPS OF PROLACTIN GENE AND ITS ASSOCIATION WITH PRODUCTION TRAITS IN NATIVE CHICKEN

Azhaguraja M*(1)

(1) ICAR- INDIAN VETERINARY RESEARCH INSTITUTE, Bareilly, Italy

*Presenting author

Corresponding author mail: azhagurajamano@gmail.com (Azhaguraja M)

The conventional method of selection based on phenotypic performance in chicken has successfully increased the quantity of production and reached the optimum level. At this stage, it is necessary to introduce the advanced technologies for the selection and breeding of poultry. Marker assisted selection (MAS) is the most efficient and advanced method in poultry breeding programme. This can be done by using different types of molecular markers namely, Restriction Fragment Length Polymorphism (RFLP), Amplified Fragment Length Polymorphism (AFLP), Randomly Amplified Polymorphic DNA (RAPD), microsatellites and Single Nucleotide Polymorphisms (SNPs). The main objective of the present study was identification 24bp indel(s) and SNPs of prolactin gene and its association with production traits in Native chicken. A total of 200 birds of Native chicken were randomly selected from All India Co-ordinated Research Project (AICRP) on Poultry Improvement farm, Mannuthy. Blood samples were collected from the randomly selected birds and isolation of genomic DNA was done by using ODP304 Origin Genomic DNA isolation kit. PCR and PCR-RFLP analysis was carried out to find a 24-bp insertion-deletion (indel) at the site of -358 of promoter 1 and two SNPs (C-2402T and C-2161G) in promoter 2, respectively of prolactin gene. The production traits viz., broodiness, age at sexual maturity, egg weight at 28, 32 and 40 weeks of age and egg number up to 40 weeks of age were measured in the randomly selected birds of Native chicken. Genotypic and allelic frequencies of polymorphic sites of promoter regions were calculated. The association between the polymorphisms of prolactin gene and production traits was determined by one way ANOVA by using the SPSS software (Version 21.0). According to the polymorphic patterns the genotypes are designated as II, ID and DD for promoter 1 and CC, CT and TT and also CC, CG and GG for C-2402T and C-2161G sites, respectively for promoter 2. All the three polymorphisms of prolactin gene were compared with production traits and found that the CC and CT genotypes of C-2402T site of promoter 2 was having significantly (p<0.05) early sexual maturity, lower egg weight and higher egg production compared to TT genotype. This result suggest that the SNP C-2402T in promoter 2 of prolactin gene can be used as a molecular marker in selection and breeding programme.
ID: 581

# GENOME-WIDE ANALYSIS OF THE GENETIC ARCHITECTURE FOR COMMERCIAL, LOCAL, FIGHT AND WILD CHICKEN POPULATIONS BY 600K SNP MICROARRAY DATA

J.Zhang\*(1), C.Nie(1), X.Li(1), Z.Ning(1), Y.Chen(2), Y.Jia(3), L.Wang(2), X.Lv(2), W.Yang(2), L.Qu(1)

(1) China Agricultural University, Beijing, China, (2) Beijing Municipal General Station of Animal Science, Beijing, China, (3) Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China

Corresponding author mail: 1599436449@qq.com (Jinxing Zhang)

After chickens were domesticated, diversified chicken populations were formed by both natural and artificial selection, which help chicken enrich abundant genetic and phenotypic variation and could be used as an ideal genetic research model. To better understand genetic architecture of chicken populations under different selection pressures, we genotyped various chicken populations with specific selection targets, including local, commercial, fighting and wild ancestral chicken breeds, by using the 600K SNP array. We analyzed the population structure, genetic relationship, inbreeding index and effective population number (Ne) and some other genetic parameters.

The wild ancestral population, red junglefowl (RJF), possessed the richest diversity with comparison to all other domesticated populations, indicated by LD, Ne and ROH results, etc. The fighting chicken breeds, which were subject to stronger male-biased selection on fighting related traits, also presented comparatively higher variation than in commercial and local breeds. After being intensely selected for reproductive and productive traits, the commercial lines showed poorest diversity. We also found the European local chicken conferred lower genetic variation than did Chinese local breeds, which could be attributed to relative shorter history of the European breeds. We also compared the effective sex-ratio of these breeds. We found commercial chickens exhibited more sex imbalance between female and male and the commercial lines showed highest values for the ratios of female to male. Interestingly, the red jungle fowl population presented more males than females.

Our results provide the population genetics information of chickens having undergone different selection pressures and could help for better conservation strategies of different chicken breeds.

Keywords: Chicken, Genetic architecture, Genetic diversity, Linkage disequilibrium, Effective population size

<footnote>\*Presenting author</footnote>
ID: 614

DIFFERENTIATION OF EXPRESSION PROFILES OF STEROL REGULATORY ELEMENT-BINDING FACTOR-1 AND -2 GENE IN CHICKEN

M.Qiu*(1), Y.Wang(2), X.Jiang(1), H.Du(1), Q.Li(1), C.Yang(1), Z.Zhang(1), C.Yu(1), H.Yin(2), Q.Zhu(2)

(1) Animal Breeding and Genetics Key Laboratory of Sichuan Province, Sichuan Animal Science Academy, Chengdu, China, (2) Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, China

*Presenting author

Corresponding author mail: mohan.qiu@163.com (Qiu Mohan)

Sterol regulatory element-binding factor-1 and -2 (SREBF1 and SREBF2) are important transcription factors involved in the regulating lipid homeostasis. Based on the essential role of SREBF1 and SREBF2, we measured the two genes mRNA expression levels in six various tissues at different growth points.

Our results shown that the SREBF1 and SREBF2 were expressed in all six tissues examined in Erlang mountainous chicken (SD02) at 42 days, and were expressed abundantly in the uropygial gland and liver, with relatively lowest levels of expression in the abdominal fat, sebum cutaneum and leg muscle. The expression ratio of SREBF1 and SREBF2 in breast muscle, leg muscle, sebum cutaneum and uropygial gland exhibit "decline-rise" trend, however, in liver, the expression ratio of these two genes exhibit "decline-rise-decline" trend. Meanwhile, the expression level of SREBF1 gene of all tissues was lower than that of SREBF2 except for uropygial gland. The findings will provide important references for further function investigation of the two genes involved in fat deposition in chickens.
ID: 1273

MOLECULAR CHARACTERIZATION OF INTERFERON ALPHA 2B GENE IN BROILER CHICKEN

D.Devara*(1), R.Chatterjee*(1), T.Bhattacharya*(1)

(1)ICAR-Directorate of Poultry Research, Rajandranagar, Hyderabad, India,
*Presenting author

**Corresponding author mail:** bhattacharyatk@gmail.com (Tarun Bhattacharya)

Interferon alpha 2B has a distinct antiviral, anti-proliferative and pro-apoptotic activities in chicken and other animals making the molecule to be therapeutically important. The objective of the study was to characterize the Interferon alpha 2B and express it in cell culture system for its therapeutic use. Blood samples were collected from a broiler chicken line (control broiler line), total RNA isolated, cDNA synthesized and the total ORF cloned pAcGFP-C1 vector. The recombinant clones were sequenced and studied for gene expression analysis in chicken embryo fibroblast primary cell culture. The sequence analysis revealed 99.31 % and 97.93 % homology at nucleotide and amino acid level between broiler and reference RJF genome. The gene sequence in broiler chicken as compared to that in reference Red jungle fowl differed at 3 positions [c. [187A>G; 399A>C; 407G>T; 454T>C]] wherein the second and third substitutions were of transversional type and the first and last ones were transitional type of mutation. However, all these nucleotide substitutions were of non-synonymous type indicating the change of amino acid (R63Q, R133S, L136R and Y152H). The isoelectric point and charge at pH 7.0 of this protein in broiler chicken was predicted as 8.954 and 8.058, while, those in reference chicken, was 8.742 and 7.225, respectively. Strongly basic, acidic, hydrophobic and polar amino acids were in the magnitude of 19, 14, 62 and 68 in reference and 20, 14, 63 and 67 in broiler protein. Secondary structure constituting alpha helix, extended strand, beta turn and random coil were in the order of 61.14, 1.55, 1.55 and 35.75 per cent in broiler and that of reference as 62.18, 1.55, 1.55 and 34.72 per cent, respectively. The tertiary structure showed mild variation in gene ontology and ligand binding. The interferon alpha 2B gene was cloned in pAcGFP-C1 vector and transfected in chicken embryo fibroblast cells. The transfected fibroblast cells revealed the expression of green fluorescent protein in fluoroscope microscope.

It is concluded that the sequence of interferon alpha 2B gene in broiler chicken differed from that of reference Red Jungle Fowl indicating non-synonymous type of nucleotide substitution with change of amino acids and the expression of this gene has been perfected in primary cell culture for its further use in combating viral diseases in chicken.

**Key words:** Interferon alpha 2B, characterization, Expression, chicken
ID: 1285

# IDENTIFICATION OF POLYMORPHISM OF PARTIAL PROMOTER OF ACACB GENE IN LAYERS

S.Chukkala*(1), R.Chatterjee(1), T.Bhattacharya(1), R.Vinoo(3), M.Muralidhar(2)

(1) ICAR-Directorate of Poultry Research, Rajendranagar, Hyderabad-500030, Telangana, India, Hyderabad, India, (2) Dept. of Animal Genetics and Breeding, N. T. R College of Veterinary Science, Gannavaram-521102. A.P, India., Gannavaram, India, (3) Dept. of Animal Genetics and Breeding, N. T. R College of Veterinary Science, Gannavaram-521102. A.P, India., Hyderabad, India

*Presenting author

**Corresponding author mail:** prasadshivach21@gmail.com (Shivaprasad Chukkala)

Genetic regulation of egg production and quality particularly, with respect to fat metabolism and quality of fat, is established based on genome-wide association studies and genome-wide gene expression studies in chicken. The chicken Acetyl CoA carboxylase beta (ACACB) gene is involved in fatty acid metabolism thereby potentially can regulate the quality of the egg.

The present investigation searched for polymorphism in the chicken ACACB gene promoter region with Single Stranded Conformation Polymorphism (SSCP) in the 5' UTR of chicken ACACB gene on two different lines of White Leghorn chickens. Genomic DNA was isolated from the blood of 250 birds of each White Leghorn lines (IWI and IWK) using standard procedures. Primers were designed for amplification of the partial promoter region of the gene. PCR-SSCP analysis was performed to determine polymorphism in the partial promoter region. Results showed that two alleles and three genotypes were present in the IWI and IWK populations. The genotype frequencies for IWI were 0.324, 0.224, and 0.452 for h1h1, h1h2 and h2h2, respectively. The genotype frequencies for IWK were found to be 0.148, 0.092 and 0.760 for h1h1, h1h2 and h2h2 respectively. Moderately higher allelic frequencies were found in h1 in IWI compare to that of IWK, whereas in IWK higher allelic frequency was found in h2 compare to that of IWI. This partial promoter of ACACB gene is polymorphic in these White Leghorn lines indicating the variability of expression ACACB gene under the control of partial promoter differential nucleotide structure.
**Physiology of nutrition**
ID: 901

# EFFICACY OF CARBOHYDRASE ENZYMES ON GROWTH PERFORMANCE OF BROILERS FED A LOW ENERGY DIET

Q.Zhang(1), H.Liu(1), J.Wu(1), M.Walsh(2), J.Berti-Sorbara*(2)

(1) DSM China Animal Nutrition Research Center, Bazhou, China, (2) DSM Nutritional Products, Kaiseraugst, Switzerland

*Presenting author

Corresponding author mail: April.Zhang@dsm.com (Qian Zhang)

A floor pen study was conducted to evaluate the efficacy of carbohydrate enzyme used alone or in combination on growth performance of broilers fed a wheat-soybean meal-sunflower meal based diet with low energy. One thousand and two hundred male Cobb 500 broilers were used in this study with a completely randomized design from 0 to 35 d of age. There were 3 dietary treatments, including: 1) control diet with the reduction of 100 kcal/kg AME and digestible amino acids (to maintain the ratios of energy to AA) as per strain recommendations; 2) control diet supplemented with 300 mg/kg RONOZYME® VP (VP, a multicomponent carbohydrate containing β-glucanase and various other side-activities such as pectinase and hemicellulase); 3) control diet supplemented with 200 mg/kg RONOZYME® VP and 100 mg/kg RONOZYME® Multigrain (MG, a multicomponent carbohydrate with the main xylanase and β-glucanases being enhanced by further side activities). The two carbohydrate enzymes were from DSM Nutritional Products. Each treatment was applied to 16 replicate pens with 25 birds per pen. Comparisons of the treatment means were performed with Tukey's test at P < 0.05, using the GLM procedure of SPSS.

Results showed that there was no significant difference in body weight gain among the treatments throughout the trial. When compared to the control group, supplementation of VP alone didn't impact the feed intake, while VP combined with MG significantly decreased (P < 0.05) feed intake at d 0-10, d 21-35 and d 0-35. Additionally, VP combined with MG showed significantly lower (P < 0.05) feed intake than VP alone at d 21-35 and d 0-35. For the feed conversion ratio (FCR), in comparison with control, VP alone significantly improved (P < 0.05) it by 4.0 and 2.5 points at d 0-10 and d 0-35, respectively; VP combined with MG significantly decreased (P < 0.05) it by 5.7, 3.1, 12.7 and 8.8 points at d 0-10, d 10-21, d 21-35 and d 0-35, respectively. The improvement of FCR from VP combined with MG was significantly better (P < 0.05) than that from VP alone at d 21-35 and d 0-35.

In conclusion, supplementation of RONOZYME® VP alone or in combination with RONOZYME® Multigrain improved growth performance of broilers fed a low energy diet, and the efficacy was more pronounced with the combination.
ID: 1158

THE DUSK FEEDING IN LAYING HENS IS SHIFTED BY LIGHT PROGRAM
VIA THE INVOLVEMENT OF CLOCK GENES

X.Wang\*(1), L.Hai(1)

(1) Shandong Agricultural University, Taian, China

Corresponding author mail: xjwang@sdau.edu.cn (Xiaojuan Wang)

Objective Dusk feeding is suggested to be probably to satisfy the energy requirement during night. Animal appetite regulation is a complex process involving interaction between the central and peripheral signals. Light sensitive suprachiasmatic nucleus (SCN) neurons synchronize peripheral clock to temporarily coordinate behavior and physiology into the environment. On the contrary, the time-dependent food intake strongly influences the molecular clock in hypothalamic SCN target areas. Hence, we hypothesized that the expression of clock genes is involved in the dusk feeding of laying hens.

Design In the present study, the feeding behavior at dust was investigated in the laying hens under two lighting programs: the normal light program (Control) with a lighting period from 05:00 to 21:00 and a dark period from 21:00 to 05:00; the shifted lighting program group (SLP) with a lighting period from 02:00 to 18:00 and a dark period from 18:00-02:00. The expressions of genes related appetite control and circadian rhythm were investigated in the hypothalamus and proventriculus at 1, 3, and 5 h before scotophase.

Results The result showed that the dusk feeding was observed from 5 h before scotophase in all the two groups. The expression of orexigenic gene neuropeptide Y (NPY) and agouti-related protein (AgRP) in hypothalamus were not changed by lighting program, whereas the expression of anorexigenic genes pro-opiomelanocortin (POMC) and cocaine-and amphetamine-regulated transcript (CART) were downregulated during dusk feeding and changed in pace with lighting program. The expression of clock genes period 2 (Per2) and cryptochrome 1 (Cry1) in hypothalamus were downregulated at dusk feeding and shifted by lighting program. In contrast, the transcriptional levels of Cry2, CLOCK, Bmal1 and Bmal2 in hypothalamus were not influenced by lighting program. In proventriculus, the expression of ghrelin was decreased during dusk feeding while the other genes measured were not influenced by lighting program.

Conclusions The result suggests that the expression of clock gene Per2 and Cry1 is linked with the downregulated expression of anorexia gene POMC and CART, and in turn the augmented feed intake at dusk. We thus conclude that dusk feeding is associated with clock gene.

<footnote>*Presenting author</footnote>
ID: 1472

EFFECT OF AQUEOUS VERNONIA AMYGDALINA ADMINISTRATION ON LYMPHOID TISSUE NUMBER IN SPLEEN AND PROVENTRICULUS OF THREE WEEKS OLD BROILER CHICKENS

R.Ofongo\*(1), B.Allen(2)

(1) NIGER DELTA UNIVERSITY, Wilberforce island, Nigeria, (2) NIGER DELTA UNIVERSITY, Wilberforce island, Nigeria

*Presenting author

**Corresponding author mail:** tariruth@live.de (Ruth Tariebi Ofongo)

Medicinal plants and spices are being highlighted as suitable alternatives to in-feed antibiotic growth promoters for sustainable poultry production without antibiotics. Plant extracts or phytogenics elicit varied response when utilized in poultry nutrition of which immune response is one. This experiment was conducted to determine the effect of aqueous Vernonia amygdalina leaf extract on lymphoid tissue number in spleen and proventriculus of three weeks old broiler chickens. A total of 150 day-old ANAAK 2000 broiler chicks were randomly distributed into three treatments having five replicates of ten birds per replicate. Birds in the control group (treatment 1) were not administered antibiotic or aqueous V. amygdalina extract. Birds in treatment 2 were administered antibiotics for 5 days. Birds in treatment 3 were administered 1ml of aqueous V. amygdalina extract on day 8 and 15 respectively. Feed and water was supplied ad libitum. One bird per replicate per treatment was slaughtered on day 15 and 21 respectively. The spleen and proventriculus was collected and placed in 10% formaldehyde prior to determining the lymphoid tissue number in each organ using appropriate procedures. The experiment was designed as a complete randomized design and lasted three weeks.

From the results obtained, lymphoid tissue number was significantly higher (p<0.05) in spleen of broiler chicks in treatment 1 (943.80 ± 96.10) and 2 (696.20 ± 188.60) respectively at two weeks compared to birds in treatment 3. Lymphoid tissue number in proventriculus was significantly lower (p<0.05) across the treatment groups.

It can be concluded that aqueous V. amygdalina extract modified immune response in the spleen and proventriculus in broiler chickens at 2 weeks of age.

**Key words:** broiler chickens lymphoid tissue, spleen, proventriculus, Vernonia amygdalina
**Protein nutrition**
ID: 438

# VALIDATION OF AMINO ACID RECOMMENDATIONS FOR BROILERS

C.Scharch(1), A.Lemme*(2)

(1) feedtest, Wettin-löbejün, Germany, (2) Evonik Nutrition & Care GmbH, Hanau, Germany

*Presenting author

Corresponding author mail: andreas.lemme@evonik.com (Andreas Lemme)

AMINOChick is a software providing amino acid (AA) recommendations for broilers. It allows the user to adjust dietary levels of digestible AA to various production conditions. The present study tested recommendations for various production objectives, for male and female broilers, respectively, varying levels of dietary energy and different degrees of whole wheat blending. Day old ROSS 308 chicks (average 36.1g/chick; 1584 male, 528 female) were allotted into 64 floor pens with 33 birds/pen and assigned to 8 dietary treatments (TRT). A 4-phase feeding program (days1-10-21-33-40) was simulated with AMINOChick: TRT1 to TRT4 followed a bifactorial design (male vs. female x maximizing body weight (BW) vs. maximizing breast meat yield (BMY)). TRT3 using males and aiming for maximizing BW served as comparison group for TRT5 and 6 in which dietary energy was reduced or increased by 2.5% in all phases, as well as for TRT7 and 8 in which phase 2 to 4 diets were blended with 5, 10, 15 and 7, 18, and 20% whole wheat. AA levels of concentrates were adjusted to energy levels and wheat inclusion as suggested by AMINOChick. While BW, feed intake, and feed conversion ratio (FCR) were recorded for all treatments, 3 entire pens from TRT1 to 4 were used to determine carcass yield (CY) and BMY. Results were assessed economically considering 18% lower costs for whole wheat compared to the wheat in pelleted diets. Recommendations for maximizing BMY resulted in higher BW, lower FCR and higher BMY (p<0.05) than for maximizing BW. Male broilers had higher BW and lower FCR than females (p<0.05) while BMY did not differ between sexes. Differentiation between performance objectives and sex was additive and was also reflected in European Efficiency Index (EEI; p<0.05). Adjusting AA concentrations to dietary energy aiming for same BW confirmed no significant differences in BW although final BW at increased energy appeared slightly increased. As expected, FCR gradually improved with increasing dietary energy (p<0.05) as did EEI. Blending diets of TRT3 with whole wheat while adjusting AA in concentrates did not affect any of the performance parameters. Income over feed cost (IOFC) was improved at highest wheat blending while dietary energy had little and indifferent effect. IOFC was higher in male than female broilers and was higher when BMY was targeted. Outcomes of the study confirmed expected effects of adjusting AA recommendations to changing production conditions and objectives.
ID: 827

# USE OF GLY AND SER IN LOW CRUDE PROTEIN DIETS CAN RESTORE THE PERFORMANCE OF BROILER CHICKENS

S.Kaczmarek*(1), M.Hejdysz(2), D.Khan(3)

(1) Poznan University of Life Sciences, Poznan, Poland, (2) Poznan University of Life Sciences, Poznan, Poland, (3) CJ Europe GmbH, Schwalbach, Germany

*Presenting author

**Corresponding author mail:** sebastian.kaczmarek@up.poznan.pl (Sebastian Kaczmarek)

The rate and efficiency of growth is lower in broilers fed diets in which crude protein has been lowered by more than 3%. The use of crystalline amino acids to formulate the lower crude protein diets for broilers has been shown to effectively reduce N excretion, improve litter quality and performance parameters.

The objective of the experiment was the identification of the minimum CP level at which the birds do not lose the performance. It was also to identify the importance of the Gly and serine level is the low CP diet. The trial was carried out with 480, 1-day old male Ross 308 chicken broilers in 5 treatments -control, and three treatments with decreased crude protein: 1, 2 or 3 pp vs. control (CP: Starter: 22, Grower: 20, Finisher:18%)and one treatment with the lowest crude protein level was supplemented with Gly to meet the Gly+Ser level of the standard diet. Each treatment was replicated 12 times with 8 birds per replicate. The total treatment duration was 35 days with a 3 phase feeding system (starter 1-11d), grower (11-28d) and finisher (28-tilt end).

Reduction in the crude protein by 1 pp did not reduce the animal performance, whereas, by 2 or 3 pp, it caused a decrease in BWG and FI and an increase in FCR (0-42nd day; P<0.05). Supplementation of diet with Gly and Ser, numerically improved BWG and FCR in comparison with control treatment. Dietary crude protein reduction affected (P<0.05) negatively carcass weight but the use of Gly and Ser improved it, resulting in a lack of differences between control treatment and Gly and Ser supplemented. Reduction of dietary protein level improved dry matter of litter (P<0.05) and reduce N excretion (P<0.05) at day 10, 24 and 35th. There was a tendency (P<0.1) to reduce blood urea nitrogen when dietary crude protein level was decreased.

Present study verifies the fact that the reduction in dietary crude protein level affects litter quality and N excretion positively. Three percent point crude protein reduction affected negatively birds' performance although Gly and Ser supplementation partially restore growth performance results to the level observed in birds fed diet with recommended crude protein level.
ID: 1353

# REDUCING DIETARY CP LEVELS IN BROILER DIETS WITH GLYCINE SUPPLEMENTATION: IMPACT ON ANIMAL PERFORMANCE AND SERUM URIC ACID PROFILE

P.Aguihe*(1), R.Omidiwura(2), A.Agboola(3), O.Odu(3), E.Iyayi(2)

(1) Federal College of Wildlife Management, New Bussa, New bussa, Nigeria, (2) University of Ibadan, Ibadan, Nigeria, (3) University of Ibadan, libadan, Nigeria

*Presenting author

Corresponding author mail: aguihepc@gmail.com (Paschal Aguihe)

**Introduction:** The supplementation of feed-grade amino acids (AA) to broiler diets has been used as a strategy to lower live production costs, nitrogen excretion and ammonia emission by reducing soybean meal inclusion that translates to lower dietary crude protein (CP) content. However, it has been observed that reducing dietary CP content may depress growth performance of broilers even with the supplementation of essential AAs, but this could be overcome by sufficient supply of the specific non-essential AA glycine (Gly).

**Objective:** The aim of this study was to investigate the effect of different CP concentrations with varied levels of Gly supplementation on performance characteristics and serum uric acid (SUA) of broiler chickens (1 - 21 d).

**Material and Methods:** 1-d old Arbor acre broiler chickens (n=360) were provided 1 of 9 dietary treatments in four replications of 36 pens (10 birds/pen) for 21 days. Three diets with levels of 22, 20 and 18g CP/kg were formulated, each containing 0.0, 0.2 and 0.4% Gly supplementation. All diets were formulated on a digestible basis, and were designed to be isocaloric. Animals and feed were weighed on days 7, 14 and 21 to determine average daily gain (ADG) and feed conversion ratio (FCR). Blood samples were collected from 2 birds per pen at d 21 to determine SUA concentrations using standard procedures. Data were analyzed by ANOVA using the GLM procedure of SAS as completely randomized design.

**Results:** Two-factor ANOVA showed significant interaction effects between CP and Gly levels for ADG (P<0.004) and FCR (P<0.005). Increasing supplemental Gly levels improved (P<0.05) ADG and FCR at various CP concentrations. Reducing dietary CP level to 18% decreased (P<0.001) ADG and increased (P < 0.001) FCR of the birds. Glycine supplementation at 0.4% resulted in improved (P<0.001) ADG and FCR compared to 0.0 and 0.2%. The SUA was observed to increase (P<0.05) at lower dietary CP level (18%) compared to 22 and 20% CP diet, while birds fed 0.4% Gly diet had the lowest (P<0.07) concentration of SUA than those fed 0.0 and 0.2% Gly diet.

**Conclusion:** This study demonstrates that higher Gly supplementation is necessary to optimize the growth performance of broilers (1-21 d) fed all vegetable based, low CP diet. Such a feeding strategy could therefore help improving the sustainability of broiler production as it is an efficient means to decrease environmental problems associated with nitrogen excretion.
ID: 1444

EFFECT OF SOYBEAN PROTEIN CONCENTRATE ON PERFORMANCE, CARCASS CHARACTERISTICS, IMMUNE RESPONSE AND INTESTINAL MICROBIAL POPULATION IN BROILER CHICKS

M.Rezaei\*(1), H.Rouhparvar(1), M.Kazemifard(1)

(1) Sari Agricultural Sciences and Natural resources University, Sari, Iran

*Presenting author

Corresponding author mail: mrezaei2000@yahoo.com (Mansour Rezaei)

This experiment was conducted to investigate the effect of different levels of soy protein concentrate (SPC) on performance, carcass characteristics, immune response and intestinal microbial population in broiler chicks. In this experiment, 160 Ross 308 chicks were used in a completely randomized design with four treatments, four replicates and 10 chicks per replicate. The treatments included control diet (treatment 1) without adding soy protein concentrate and diets containing 10% (treatment 2), 20% (treatment 3) and 30% (treatment of soy protein concentrate replaced by the protein provided by soybean meal).

Results showed that, use of experimental treatments had significant effect on body weight gain and feed intake in starter, grower and finisher period of the experiment (P <0.05). The effect of experimental treatments on feed conversion in grower, finisher and whole periods was significant (P <0.05). Treatment 2 showed the best weight gain, feed intake and feed conversion ratio. Breast percentage was higher in treatments 2 and 4 in comparison to other treatments (P <0.05). The results also showed that the effect of experimental treatments on duodenum length was significant (P <0.05). The duodenum length decreased with increasing soy protein concentrate in the diet. The effect of experimental treatments on dry matter, crude protein, crude fat and crude ash digestibility was significant and treatments 2 and 3 had better digestibility for dry matter, crude protein and crude fat (P <0.05). The effect of experimental treatments on gizzard pH content was significant (P <0.05). Treatments 2 had the lowest pH. The effect of experimental treatments on litter moisture, antibody titre against sheep red blood cells, intestinal microbial population and duodenal morphology was not significant.

Results of the present experiment showed that replacing 10% of soybean meal protein by the soy protein concentrate improved the performance and nutrients digestibility in broiler chicks.

Keyword: Carcass Characteristic, Immune Response, Junior Broiler Chicken, Microbial Population, Soybean Protein Concentrate.
# DETERMINING STANDARDIZED ILEAL DIGESTIBLE AMINO ACID AND DIETARY ENERGY REQUIREMENTS IN BROILER BREEDERS USING THE NITROGEN BALANCE METHOD

J.Weil*(1), A.Beitia(2), N.Suesuttajit(1), K.Hilton(3), P.Maharjan(1), D.Martinez(1), J.Caldas(4), S.Rao(4), C.Coon(1)

(1) University of Arkansas, Fayetteville, United states, (2) University of Arkansas, Fayetteville, United states, (3) University of Arkansa, Fayetteville, United states, (4) Cobb-Vantress, Inc., Siloam springs, United states

*Presenting author

Corresponding author mail: jtweil@uark.edu (Jordan Weil)

Inclusion rates of amino acids in poultry feeds are dependent on economics related to performance and primary breeder recommendations. Considering dietary programs have a direct effect on hatching eggs, the amino acid requirements of broiler breeder hens should be continually updated to ensure the hen is reaching its maximal genetic potential. The standardized ileal digestible (SID) AA requirements of egg mass can be determined to predict more accurate requirements for broiler breeders (BB). Additionally, the dietary energy and maintenance requirements can be determined in order to ensure birds are not receiving excess calories. In this experiment, the nitrogen balance technique was used to determine the AA requirements for the Cobb 700 FF BB during two important production periods. A total of 648 21 wk-old Cobb 700 FF BB were placed in metabolic cages to study eight test AA (10 reps/diet) (IACUC #19028). Eighty-eight birds served as control. Ten replicates were used to determine SID requirements. Test AA included methionine and cysteine (Met + Cys), lysine (Lys), arginine (Arg), valine (Val), isoleucine (Ile), threonine (Thr), tryptophan (Trp), and glutamic acid (Glu; SID protein). Birds were fed for a period of 7 weeks (2 wk adaptation; 5 wk testing) in two phases. Phase I (PI) lasted from wk 30 through 37, while Phase 2 (PII) occurred from wk 44 through 51. The egg mass produced by breeders was evaluated in each period.

Results were analyzed using a quadratic model (JMP Pro 14.1). For egg mass, SID requirements for Met+Cys, Lys, Arg, Ile, Val, Thr, and Trp were 880, 900, 900, 787, 767, 672, 256 mg/day, respectively, for PI, and 803, 1110, 933, 811, 811, 681, and 235 mg/day, respectively, for PII. Glu (SID protein) requirements were 20 and 19 g/day, respectively, for PI and PII. The digestible AA requirements were comparable to previous digestible AA requirements determined for Cobb 500 FF hen at peak production (Ekmay, 2013).

Knowledge gained from this study will provide the necessary information to determine the production requirements of Cobb 700 hens.
Quantitative genetics
ID: 667

STUDYING THE RESPONSE TO FAMILY BASED SELECTION AMONG
DIFFERENT LINES OF JAPANESE QUAIL THROUGH THREE
GENERATIONS

J.Hussain*(1), A.Mahmud(1), A.Rehman(1), A.Ghayas(1), S.Mehmood(1), S.Ahmad(1)

(1) University of Veterinary and Animal Sciences Lahore Pakistan, Lahore, Pakistan, (

*Presenting author

Corresponding author mail: jibran.hussain@uvas.edu.pk (Jibran Hussain)

Recently, in Pakistan, Japanese quails (Coturnix Coturnix Japonica) locally called as "BATAIR" have made significant presence in the meat markets, however, at present, the major problem regarding quail production is poorer body weight gain, feed efficiency, and reproductive performance, and its size at a specific age (04 weeks) is not large enough to serve at dining tables; reason being very little research work conducted on such birds.

Keeping in view such situation, a project was planned to improve growth performance of broiler quail and productive and reproductive potential of the parent stock of Japanese quail through family based selective breeding program. For this purpose, it was planned to build 04 lines, each having specific traits to be focused for selection in each generation. Line I was assigned higher body weight at four weeks of age with improved FCR, line II with higher body weight at four weeks of age and improved egg number till 14 weeks of age, line III with higher body weight at four weeks of age and the males having large foam gland size, while, line IV was assigned higher body weight at four weeks of age and increased number of holes in vitelline membrane through sperm penetration. Family based selection was performed with only the families having above average performance were selected to get the next generation. Data were collected through three generations (0, 1 and 2). Two hatches were completed in each generation in each line. Incubation, brooding, growing and laying management conditions were the same for all birds in all lines. Statistical analysis of data indicates that bodyweight base line had higher growth performance and carcass cut-ups yield in successive generations. The response to selection practices indicates that there is much more potential to be explored for body weight. In addition, egg production line of quails had higher egg production in G2 than G1 and G0 and better FCR per dozen eggs and per unit of egg mass that can be very helpful for getting next progeny. In selection for bigger cloacal gland and pores on vitelline membrane, Japanese quails presented the highest fertility and hatchability during 2nd generation than G0 and G1. All four lines improved the assigned characters significantly from G0 to G2. It can be concluded that there is enough potential in the existing population of Japanese quail for higher body weight, higher egg production, as well as fertility and hatchability.
ID: 739

# BEST LINEAR MODEL AND CORRELATION BETWEEN BODY WEIGHT AND BODY LINEAR MEASUREMENTS OF THE FRENCH BROILER GUINEA FOWL RAISED IN THE SUB-HUMID TROPICS OF NIGERIA

J.Dzungwe* (1), G.Deeve sabastein(2), E.Joseph(2)

(1) Regional Center of Excellence for Avian Sciences, University of Lome, Togo, Lome, Togo, (2) Federal University of Agriculture, Makurdi, Makurdi, Nigeria

*Presenting author

**Corresponding author mail:** torsagain@gmail.com (Joshua Torkuma Dzungwe)

Genetic and phenotypic correlations are useful in predicting responses to selection and determining optimum weight when improving more than one trait. Body linear measurements provide information on performance, productivity and carcass characteristics of animals. There is paucity of reported work on body linear measurements of the French broiler guinea fowl in Nigeria. This study was to determine the best linear model and estimate the relationship between body weight and body linear parameters of the French broiler guinea fowl in Nigeria. The study was conducted in Makurdi, Benue State, Nigeria. The experimental design was complete randomized design. 86 French Broiler Guinea Fowls were used for the experiment, they were housed in a dwarf wall wire mesh house with corrugated roofing sheets on a deep litter system and fed commercial grower diet containing 18% crude protein. Feed and water was provided ad libitum. Data was collected at 26 – 39 weeks of age. Body weight (BW) was measured using sensitive weighing scale and body conformations like shank length (SL), thigh length (TL), keel length (KL), wing length (WL), body length (BL) and chest circumference (CC) were measured using flexible measuring tape. Data was subjected to Minitab 2003 for analysis to generate descriptive statistics and estimate the correlation between body weight and body linear measurements; it was also subjected to simple and multiple regression analysis. Results showed that there exist positive low-moderate phenotypic correlations between body weight and shank length (0.437), thigh length (0.397), keel length (0.044), chest circumference (0.409), body length (0.390) and wing length (0.374). Shank length and chest circumference showed the highest correlation coefficients. The regression models for body weight prediction using body linear measurement on body weight showed increased prediction strengths from 19-29.9% as the exponents (SL,TL, KL, CC, BL and WL) increased. The regression model combining all body linear measurements (BW=104.15+0.18SL+0.18CC+0.20TL+0.18BL) predicted body weight with highest accuracy (29.9%). Genetic improvement for body weight of the French broiler guinea fowl populations in Nigeria can be achieved by selection for shank length and chest circumference. The regression model for combined body linear traits can be used effectively to predict body weight of the French broiler guinea fowl in areas where weighting scale is not readily available
ID: 943

GENETIC BASIS OF WOODEN BREAST AND WHITE STRIPING IN
COMMERCIAL BROILERS

J.Lake*(1), M.Walugembe(2), L.Kramer(2), J.Dekkers(2), B.Abasht(1)

(1) University of Delaware, Newark, de, United states, (2) Iowa State University, Ames, ia, United states

*Presenting author

Corresponding author mail: dnovick@udel.edu (Juniper Lake)

Wooden breast (WB) and white striping (WS), the two major myopathies of fast-growing broilers, continue to threaten global poultry production due to their severe impacts on meat quality. Tightly associated with economic traits such as growth rate, feed efficiency, and breast muscle yield, these muscle disorders present an exceptional challenge to producers, as dietary or management strategies against WB and WS generally impair performance. An understanding of the genetic basis of these myopathies is therefore critical for developing a long-term solution. To this end, a genome-wide association study was conducted using 1,193 Cobb 500 broilers, which were raised to seven weeks of age and evaluated for WB, WS, and body weight. Genotyping was performed using restriction enzyme-based genotyping by sequencing and quantitative trait loci were detected using single-SNP analysis and Bayesian multi-marker regression (BayesB).

Heritability was estimated to be moderate for all traits: 0.49±0.07 for WB, 0.50±0.06 for WS, and 0.40±0.06 for body weight. Genetic correlation between WB and WS was high (0.88±0.04), but genetic correlation between either WB or WS and body weight was low (0.15±0.11 and 0.08±0.11, respectively). Multi-marker analysis of WB found ten 1-Mb regions that each explained >1% of genetic variance. Together, these ten regions explained 18.4% of the genetic variance for WB. Multi-marker analysis of WS and body weight identified, respectively, four and zero 1-Mb regions that each explained >1% of genetic variance.

These results improve our understanding of the genetic architecture of
these myopathies and suggest that WB and WS would respond to
selection.
ID: 945

ESTIMATION OF GENETIC PARAMETERS FOR BEAK SHAPE IN
COMMERCIAL TURKEYS (MELEAGRIS GALLOPAVO)

H.Emamgholi begli*(1), S.Shadpour(1), H.Dalton(2), S.Torrey(1), D.Tulpan(1),
A.Harlander-matauschek(1), C.Baes(1), B.Wood(3)

(1) University of Guelph, Guelph, Canada, (2) Compassion in World Farming, Georgia, United states, (3)
University of Queensland, Queensland, Australia

*Presenting author

Corresponding author mail: emamgholi202@gmail.com (Hakimeh Emamgholi Begli)

Various management strategies can be applied to reduce the incidence of
feather pecking and cannibalism in poultry, one of which includes beak
treatment. In group housing systems, flocks with shorter, blunter beaks
(via beak treatment) have better feather quality and lower mortality.
Therefore, the objective of present study was to estimate the genetic
parameters for beak shape, length, width, and depth in commercial
turkeys using geometric morphometrics. Right lateral images of 1470
turkeys at 6 weeks of age were used. Landmarks of the upper mandible
shape were digitized using the tpsDig morphometric software for each
turkey. Beak shape classes were clustered into three groups using K-
means clustering with the aim of increasing variation between groups
and decreasing variation within each group. Sex and an animal direct
genetic effects were included in the statistical model. Gibbs sampling was
used to obtain the parameters of interest for the beak shape traits.
Heritability estimates for beak shape, length, width and depth were
0.169 ± 0.046, 0.145 ± 0.047, 0.128 ± 0.040 and 0.367 ± 0.093,
respectively. Length was positively genetically correlated with width (rg=
0.953 ± 0.028) and negatively genetically correlated with depth (rg= -
0.458 ± 0.29). Width had a negative genetic correlation with depth (rg=-
0.542± 0.291). The genetic parameters of the studied traits suggest that
beak shape is heritable. Further analyses should be conducted to better
understand the association between beak shape and feather pecking
behaviour, as well as to estimate phenotypic and genetic correlations with
other traits of economic and animal welfare importance for the turkey
industry.
Reproductive biotechnologies
ID: 1082

# PRIMARY ASSESSMENT OF BANKABILITY OF RARE NATIVE CHICKEN BREED AS CRYO-PRESERVED PGCS: CHICKEN-CHICKEN VS CHICKEN DUCK TRANSFER SYSTEM

A.Naim*(1), A.Naim*(2), S.Dash(1), S.Mishra(3), D.Nayak(4)

(1) KALINGA INSTITUTE OF INDUSTRIAL TECHNOLOGY - TECHNOLOGY BUSINESS INCUBATOR, Bhubaneswar, India, (2) KIIT-Technology Business Incubator,, Bhubaneswar, India, (3) Central Avian Research Institute,, Bhubaneswar, India, (4) Discipline of BioSciences and Biomedical Engineering,, Indore, India

*Presenting author

Corresponding author mail: adnan.kiittbi@gmail.com (Adnan Naim)

Bio-banking of avian germplasm has opened an opportunity for the conservation of indigenous rare chicken breeds which bears highly valuable phenotypic and genetic traits and characters. Successful use of Cryo-preservation of early embryonic cells like Primordial Germ Cells (PGCs) have been documented by many researchers, worldwide, for conserving germplasm of domestic, rare and endangered avian species. In-vitro propagation of cryo-preserved PGCs thus emerges as the trusted options for bio-banking as well as reconstitution through germline chimerism, for breeds of importance. We have attempted to bio-bank the germplasm of 2 important native chicken breeds of India, using both intra-species and inter-species PGC transfer systems. Our primary objective was to study and compare the practicality of germplasm bankability of donor PGCs, as attempted through the usage of harvested-PGCs (chicken origin) into regular hosts like White Leghorn (WLH) (intra-species) vs PGC transfer from native chicken breeds to an extra-species (duck) recipient system. We have harvested the blood and gonadal PGCs of Aseel, Kadaknath and White Leghorn (WLH) and further cultured them in-vitro and later cryo-stored them in liquid nitrogen. Upon revival of PGCs, we have transferred them into the White Leghorn and Duck to assess the practicality of acceptance of PGCs intra-species or inter-species that can develop fertile chimeras to regenerate the pure-line breed. The results show that Aseel and Kadaknath PGCs successfully colonized into the gonads of the recipients (intra-species; WL and inter-species; Duck) with no significant difference.

This suggests that the Duck embryo didn't show any immune response in the form of rejection of exogenous PGCs and lead to the development of the embryo. This also implicates that indigenous chicken germplasm can be bio-banked efficiently in the same species host as well as different. Based on the results, our future experiments are focused on re-constituting the indigenous chicken breed by generating germline chimeras.
Reproductive physiology
ID: 214

# A PHYTOGENIC FEED ADDITIVE TO IMPROVE MALS AND FEMALE REPRODUCTIVE PERFORMANCE AND DAY-OLD CHICK QUALITY

M.De Laet(1), M.Osredkar Mergole(1), R.Montanhini Neto*(1)

(1) Delacon Biotechnik GmbH, Engerwitzdorf, Austria

*Presenting author

Corresponding author mail: manu.delaet@delacon.com (Manu De Laet)

Male fertility is of utmost importance for breeder flock performance as it is directly related to the number of day-old chicks per hen housed. One of the key factors for male fertility is sperm quality. This is related to an effective antioxidant system in the spermatozoids to reduce the concentration of intracellular free radicals that induce sperm DNA fragmentation and lipid peroxidation in the sperm cell membrane. Phytogenics can have direct (via polyphenols) and indirect antioxidant effects. The mode of action of the latter is via the Nrf2- pathway upregulating the nuclear antioxidant response element (ARE), which results in increased production of anti-oxidant enzymes. Furthermore, essential oils and saponins can increase testosterone and semen production. Biostrong® Fertile (BSF) is a proprietary blend of essential oils, flavonoids, and quillaja saponins developed to improve male breeder fertility.

The efficacy of BSF was tested in Zootests, Ploufragan, France in a breeder flock between 43 and 53 weeks of age. A total of 54 individually housed male breeders (Hubbard) were allocated to each treatment and fed either a control diet or control diet supplemented with 750 g/t BSF (from week 43 weeks on). Males were milked for artificial insemination. Females (9 pens per treatment with 24 birds per pen) were inseminated with semen of the control or the BSF group. At 53 weeks of age, in total 1950 eggs per treatment were collected and set to determine fertility, hatchability and first quality day-old chicks (DOC: % of hatched chicks). Day-old chicks of both groups were followed during the first week. Fertility rate was significantly (P<0.01) increased from 83.8 to 88.5%, hatchability rate increased from 93.7 to 95.5% and first quality DOC rate increased from 95.7 to 96.1% at 53-weeks of age. Per 100 eggs set, the number of DOC was increased by 5.7 in 53-week old breeders through supplementation of male breeder diets. In a separate trial it was also shown that BSF increased testosterone levels in male breeders (+ 34.9%), which could account for the higher semen production. Day-old chicks from BSF group showed also a higher body weight at 7 days of age, 96.4 g for BSF versus 93.7 g for control.

It was concluded that dietary supplementation of male broiler breeder diets with Biostrong® Fertile (a phytogenic feed additive) resulted in higher female reproductive performance and higher DOC performance.
ID: 1179

# TRANSCRIPTOME SEQUENCING REVEALS CANDIDATE GENES FOR EGG RATE DURING PEAK LAYING PERIOD AND END-OF-LAY HENS

E.Hao\*(1), H.Chen(1), R.Huang(1)

(1) Hebei Agricultural University, Baoding, China

\*Presenting author

**Corresponding author mail:** 277179119@qq.com (Erying Hao)

## Introduction

Laying hens have a sharp drop in laying rate when ovarian follicles are exhausted. However, low laying rate in aging laying hen is a result of a dynamic interaction between neuroendocrine changes that take place in the brain with the reproductive endocrine axis governing the function of ovaries.

## Material and Methods

To elucidate the key regulatory genes involved in Dawu Jinfen chicken with high and low rates of egg production, RNA from hypothalamus tissue during 35-week-old high laying rate 100%, 100%, 100% (Group YH:YH1, YH2 and YH3), 70-week-old high laying rate 100%, 98%, 100% (Group OH: OH1, OH2 and OH3) and 70-week-old low laying rate 55%, 45%, 30% (Group OL:OL1, OL2 and OL3) were sequenced using the Illumina HiSeq 2500 sequencing platform.

**Results:** A total of 391.59 million clean reads with an average of 43.51 million reads per sample (range: 40.04 to 45.71 million) were acquired after removing the low-quality and adaptor sequences. A comparison between Groups OH and OL revealed 32 differentially expressed genes, with 15 displaying up, and 17 displaying down, A comparison between Groups OH and YH revealed 56 differentially expressed genes, with 30 displaying up, and 26 displaying down, A comparison between Groups OL and YH revealed 127 differentially expressed genes, with 30 displaying up, and 97 displaying down. Ten genes that are mainly involved in pathways for reproduction regulation, such as PRLH, ESR2, CGA, CRH, CYP19A1, CYP26B1, PGR, OXT. All of the predicted genes of them were mapped to terms in the GO and KEGG databases. In total, in OH-VS-OL groups, 2 GO terms and 55 KEGG pathways in hypothalamic tissues were significantly enriched.

## Discussion and conclusion

The hypothalamus is a key component of the HPG system. As the tip of the HPG axis, it plays an important role in hormone secretion and regulation of reproductive performance in laying hens. As we expected, our results aggregated many reproductive-related signaling pathways, such as the GnRH signaling pathway, the estrogen signaling pathway, and the prolactin signaling pathway. The obtained RNA sequencing data from the hypothalamus could be a valuable resource for genomic studies and provide new insights into the biological processes that regulate chicken reproduction. These data provide comprehensive gene expression information at the transcriptional level that might increase our understanding of the laying hens' reproductive biology.
ID: 1266

EXPRESSION OF INHIBIN A (INBHA) GENE IN THECA CELLS DURING LATE LAYING STAGE IN WHITE LEGHORN LAYER CHICKEN

Latha Preethi Andrew*(1), R. Chatterjee*(2), P. Kumar(1), C. Paswan(2), M. Dange(2), T. Bhattacharya*(2)

(1) ICAR-Indian Veterinary Research Institute, Bareilly, India, (2) ICAR-Directorate of Poultry Research, Hyderabad, India

*Presenting author

**Corresponding author mail:** bhattacharyatk@gmail.com (Tarun Bhattacharya)

Inhibin is an endocrine negative regulator of follicle stimulating hormone (FSH) release from the anterior pituitary gland in chicken and other animals. It also has antagonistic effect on the activin receptors viz. ActRIB, ActRIIA and ActRIIB and thus, impedes the recruitment of activin receptors and induction of the intracellular activin signaling cascade. Inhibin has two sub-units forming the heterodimer to form the functional inhibin molecules. Inhibin alpha alongwith beta-A subunits form inhibin A (INHBA) in chicken. The objective of this study was to determine expression of INHBA in the theca cells of the ovarian follicles in a White Leghorn chicken line (IWF).

The mRNA expression of the gene was studied with realtime PCR where the primer pair was designed encompassing intron 1 and the primers used in this study were INHAEXPF (5'-GGACAATTGGATCGTGCAC-3') and INHAEXPR (5'-CACCGTCTCGTACTTGAAG-3'). Three biological replicates from IWF White Leghorn chicken line at 64 weeks of age were selected randomly for this study. Two genes viz. beta-actin and GAPDH were used as internal control. The real time PCR study was carried out in triplicates for each sample and each gene. Accordingly, average ct value obtained for activin and GAPDH was estimated as 20.1. The mRNA expression of INHA was observed in the theca cells of IWF chicken at 64 weeks of age. The expression in terms of 40-DeltaCt varied from 32.9 to 38.4. The CV with respect to variation of INHBA expression among the birds was 8.54% indicating homogeneity of expression of this gene among the birds.

From the results, it is concluded that inhibin A expression was detected in theca cells at advanced egg laying stage in White Leghorn chicken line indicating its immense role in egg production.

**Keywords:** Egg laying stage, Gene expression, Inhibin A, Layer chicken, Theca cells
Small scale family poultry farming
# THE IMPACT OF POULTRY INFORMATION ECONOMICS ON STRENGTHENING THE COMMERCIAL AND TECHNICAL CAPACITIES OF SMALL-HOLDER POULTRY FARMERS IN SOUTH AFRICA.

H.Swatson*(1), K.Thapelo(2), T.Sithole(3)

(1) Elsenburg Agricultural Training Institute, Cape town, South Africa, (2) Elsenburg Agricultural Training Institute, Oudtshoorn, South Africa, (3) Cedara College of Agriculture, Pietermaritzburg, South Africa

*Presenting author

**Corresponding author mail:** swatsonhk@gmail.com (Harry Swatson)

Smallholder poultry production continue to be an integral part of farming systems, socio-cultural and household economies. Agricultural and agricultural related skills development along the whole value chain is a core element of support and empowerment for smallholder poultry farmers. Training per se may not have the desired impact if the smallholder poultry farmer is not practically guided on how to smartly apply the acquired knowledge and skills acquired. The objective of this paper will be to explore the impact of poultry production information economics on small-holder poultry farmers. Responses from 482 farmers were collected. The primary research methodology involved workshops, focus group sessions, surveys, and interviews.

Results- Key challenges of smallholder poultry farmers include the limited poultry skills, fragmentation of supply of poultry products, the inefficient integration into poultry value chains, inadequate production and processing technologies, and the lack of an adapted financial services to meet their needs. Eighty-five percent of the respondents do not benefit from upstream poultry business opportunities. Sixty five percent were aware of the prescribed standards regarding the quality, class or grade, packing, marking and or labelling requirements for the sale of eggs. Ninety percent of farmers indicated that poultry training on aspects such as chick management, environmental control, feeds and feeding management, and disease control and prevention did lead to practices that enhanced their poultry farm performance. All farmers gave an indication that production and market access alone cannot make smallholder poultry farmers successful.

There is a steady growth of interest in making use of poultry production as a tool in improving the socio-economic livelihoods of rural households. Caution must be exercised with the introduction of imported dual purpose breeds into the traditional backyard poultry rearing systems (Swatson and Themeli, 2012). Poultry training, coaching and mentorship are important. The impact of poultry information was observed to strengthen the commercial and technical capacities of smallholder farmers.

## Ref:

Swatson, H. and Themeli, R. (2012) Egg production and quality characteristics of the Venda and Ovambo chickens kept under improved rearing systems in KwaZulu-Natal South Africa. WPSJ Supplement 1, WPC 2012. 5-9 August. Brazil.
ID: 1018

EMPOWERMENT OF RURAL WOMEN THROUGH NATIVE CHICKEN REARING UNDER TROPICAL CLIMATE

Ezhil Valavan Subbiah*(1), Pandian.C.(1), Selvan..S.T.( 1), Sundaresan.A.( 1), Omprakash.A.V.( 1)

(1) TAMILNADUVETERINARY AND ANIMAL SCIENCES UNIVERSITY (TANUVAS), INDIA, Chennai, India, (2) Tamil Nadu Veterinary and Animal Sciences University, Chennai, India

*Presenting author

Corresponding author mail: vetezhil2015@gmail.com (Ezhil Valavan Subbiah)

Native chicken rearing is gaining momentum now among jobless youth, school dropouts, women, etc., in rural areas. Majority of the landless or farm women are keeping 10-15 non-descriptive native chicken under back yard rearing. Native chicken rearing can be easily adopted even by illiterates with much less skill. Native chicken are more sturdy and well adapted to local climatic conditions, can be sustained entirely in free range and hand feeding. The eggs and meat of native chicken fetches high premium due to high consumer acceptability even in the urban and semi-urban sectors where plenty of eggs and poultry meat from commercial units are available. National Bureau of Animal Genetic Resources of India has recognized 19 distinct native breeds of chicken. With the above facts, the present work was undertaken to analyse the status of farm women in native chicken rearing. A data on native chicken rearing was collected from 300 farm women of various districts Tamil Nadu. The information on management practices of native chicken production was analyzed. All the data were collected by personal interview method using a standard pre-tested questionnaire specially designed for the purpose.

Majority of the women (80 per cent) involved in native chicken rearing was belonging to young (20-35 years) age group. The average flock size reported in the study area was 15-20 birds. Majority of the farm women (82 per cent) were rearing native chicken under free range during day time and provide shelter during night time. Rice was the major source of feeding, along with cereal grains and kitchen waste. Most of the farm women (95 per cent) were not vaccinated against any disease. The average selling price of the meat is ± 4.28 / Kg, when it attained a body weight of 1.00 -1.25 kg with a cumulative feed efficiency of 3.0 -3.2. Livability is about 90-94 per cent. The adult birds reared up to 3 years for breeding with an average of 80 eggs produced per female per year. The average selling price of an egg is ± 0.21.

Hence, the native chicken rearing will provide subsidiary income to rural families and improve nutritional status of the family.
Turkeys
ID: 1242

HIGH LEVELS OF PHYTASE AND DIFFERENT WATER PH ON PERFORMANCE, METABOLIZABILITY AND PLASMA INOSITOL OF TURKEYS

L.Schmidt Bassi(1), L.Teixeira*(2), A.Maiorka(1), R.Fernando sens(3), J.W. wilson(4), S.Etheve(4)

(1) Federal University of Paraná State, Curitiba, Brazil, (2) DSM Nutritional Products, Mairinque, Brazil, (3) DSM Nutritional Products, Curitiba, Brazil, (4) DSM Nutritional Products, Kaiseraugst, Switzerland
*Presenting author

Corresponding author mail: levy.teixeira@dsm.com (Levy Do Vale Teixeira)

The aim of this study was to evaluate the effects of different water pH on performance, metabolizability and plasma inositol of turkeys fed high dietary levels of phytase. A total of 560 one-d-old male turkeys were distributed in a completely randomized design in a 3x2 factorial design: 3 doses of phytase (0, 1000 and 2500 FYT/kg) and 2 water pH (5.5 and 8.2), with 6 treatments and 7 replicates of 10 birds. Birds received feed based on corn-soybean meal, and water ad libitum. Water pH levels were obtained by adding NaOH solution (for pH 8.2) and KCl solution (for pH 5.5) in due proportions to drinkable water. Performance was measured by feed intake (FI), body weight gain (BWG) and feed conversion ratio (FCR) from 1 to 21 days. Partial excreta samples were collected daily from 19 to 21 days, freeze-dried and grounded for analysis and calculation of the apparent metabolizability coefficient of dry matter (AMCDM), mineral residue (AMCMR), crude protein (AMCCP), calcium (AMCCa), phosphorus (AMCp) and apparent metabolizable energy (AME). At 21 days, 2 birds per repetition were selected for blood samples collection, which were centrifuged to separate the plasma. Plasma concentration of myo-inositol (MYO) was determined by mass spectrometry using an UPLC-TQD system.

There was no interaction among treatments for performance and MYO (P>0.05), which were only affected by phytase doses. Inclusion of 1000 and 2500 FYT/kg resulted in higher BWG (625.9 g and 634.1 g respectively) and better FCR (1.60 and 1.59 respectively) compared to control diet (WG=589.8 g; FC=1.66) (P<0.05). MYO was linearly (P<0.05) increased by phytase inclusion (232.2 and 236.3 µmol/L for 1000 and 2500 FYT, respectively) compared to unsupplemented diet (186.8 µmol/L). An interaction was observed for phytase and water pH for AMC variables (P<0.05). The best results of AMCMR (47.7%) and ADE (3496 kcal) were obtained with 2500 FYT/kg and 5.5 water pH, whereas AMCp increased with phytase inclusion regardless of water pH, up to 61.6%. Separately, 2500 FYT/kg dose significantly increased AMC of DM, CP and Ca up to 75.1, 65.8 and 45.66%, respectively (P<0.05). Water pH of 5.5 increased AMCCP (P<0.05).

In conclusion, the supplementation of turkey diets with high levels of phytase improves performance and MYO absorption, and when offered with an acidified water supply, nutrient metabolizability can be enhanced.
Welfare and behaviour
ID: 1020

# NOVEL BIOMARKERS TO TEST FOR POULTRY WELFARE

T.Crowley*(1), M.Craven(1), P.Taylor(2)

(1) Deakin University, Geelong, Australia, (2) University of New England, Armidale, Australia

**Corresponding author mail:** tamsyn.crowley@deakin.edu.au (Tamsyn Crowley)

Heavily driven by public perception welfare is of major concern for the poultry industry. The past few decades have seen development of poultry practices with a heavy focus on the welfare of poultry. There have been huge changes implemented to accommodate improvements in welfare, particularly in the layer industry, but what is driving this change? Some argue it is supermarket monopolies; others suggest it is people's perceptions of hen welfare. This drive has highlighted the need to develop definitive tools that will enable us to accurately assess hen welfare.

Here we present a new suite of novel scientific biomarkers (microRNAs) that may be used to determine the current affective state of laying hens. We have focused on both positive and negative affective states, namely reward and stress states. Our studies have developed innovative animal models to assess positive and negative affective states. From these studies we have detected biomarkers in plasma that accurately define the two affective states. In addition, we have confirmed our findings using traditional methods of assessing welfare namely, behaviour assessments and hormone measurements. Our tests have been developed to interrogate pooled samples rather than an individual hen in an effort to more accurately assess commercial flocks. Subsequently we have tested a range of different housing systems used in Australia (caged, barn and free range) to determine the levels of stress present in these systems. The sensitivity of our tests has been confirmed through testing blind samples provided directly from industry.

In conclusion, we present a novel biomarker test that can reliably be used to assess hen welfare using pooled blood samples. We are currently working on methods to explore these biomarkers in non-invasive samples such as eggs.

<footnote>*Presenting author</footnote>
ID: 987

KEEL BONE FRACTURES AFFECT THE MOVEMENT PATTERNS AND NUMBER OF DUST BATHING BOUTS OF AVIARY HOUSED LAYING HENS

V.Witjes*(1), Y.Gomez(2), F.Sousa(3), T.Rodenburg(4), M.Toscano(2)

(1) Graduate School of Life Sciences (GSLS), Utrecht University/Center for Proper Housing: Poultry and Rabbits, Animal Welfare Division, Veterinary Public Health Institute, University of Bern, Zollikofen, Switzerland, (2) Center for Proper Housing: Poultry and Rabbits, Animal Welfare Division, Veterinary Public Health Institute, University of Bern, Zollikofen, Switzerland, (3) Veterinary Public Health Institute, University of Bern, Liebefeld, Switzerland, (4) Department of Animals in Science and Society, Faculty of Veterinary Medicine, Utrecht University, Utrecht, Netherlands

Corresponding author mail: vivian.witjes@vetsuisse.unibe.ch (Vivian Witjes)

Keel bone fractures (KBFs) frequently occur in aviary housed layers and have been shown to relate to less time spent in the litter and more on the top tier. This shift in location duration may be related to hens with KBFs avoiding higher local density in the litter. For instance, the hens' reduced mobility may result in less time engaged in synchronised behaviours, such as dust bathing (DB), a highly motivated behaviour for laying hens, and piling, a dense accumulation of hens in the litter. Furthermore, since KBFs are suggested to be painful, hens may show less DB and piling in order to avoid pressure exerted on the keel during both behaviours. In this study, it was investigated whether laying hens with KBFs: 1) spend less time in the litter and more time on the top tier and 2) spend less time on DB and/or piling when in the litter area. The mobility of 68 hens was assessed in a semi-commercial pen with a total of 225 birds, by using an infrared tracking system allowing data on the duration within- and transitions between tiers and litter to be collected from 32-39 weeks of age. Keel bone health of all tracked hens was repeatedly assessed by palpation, while DB and piling of 41 of the tracked birds was measured by live observations.

All data were analysed by (generalised) linear mixed effect models. Hens with KBFs spent less time in the litter area (with KBFs: 305±9 min; without KBFs: 448±6 min; P=0.02, n=68) and more time on the top tier (with KBFs: 346±13 min; without KBFs: 121±9 min; P=0.01, n=68). However, no effects on the percentage of time engaged in DB (overall mean: 18.3±1.1%, P=0.83, n=41) and piling (overall mean: 5.7±0.5%, P=0.51, n=41) when in the litter were found. Although KBFs did not influence the relative DB duration, hens with KBFs performed fewer DB bouts (with KBFs: 1.6±0.1; without KBFs: 2.0±0.1; P=0.04, n=41). Thus, compared to hens without KBFs, hens with KBFs do not seem to avoid dense groupings in the litter and remain motivated to perform DB, but may alter their daily behavioural pattern of DB as reflected by fewer bouts, perhaps due to pain experience.

Taken together, this study reveals that KBFs are negatively associated to movement patterns and number of DB bouts, and therefore are a likely welfare issue for aviary housed laying hens.

<footnote>*Presenting author</footnote>
ID: 280

A COMPARISON OF THREE PEN STUDIES INVESTIGATING THE EFFECT
OF PERCHES ON ACTIVITY AND MOBILITY IN THE MODERN MEAT
CHICKEN SPECIMEN.

D.Phibbs*(1), P.Groves(1), W.Muir(1)

(1) The University of Sydney, Sydney, Australia

*Presenting author

Corresponding author mail: danielle.phibbs@sydney.edu.au (Danielle Phibbs)

Perches are a form of environmental enrichment that are being
increasing utilised in modern meat chicken rearing environments.
Significant research has been published addressing whether they pose
any behavioural or physiological benefits to birds, but results have been
widely inconsistent. This paper takes into account three comparable pen
studies performed at the University of Sydney, in an attempt to
identify patterns in perch use and the perch effect on activity and
mobility. In a series of three studies, day old Cobb 500 birds were
placed in pens with or without perches, to observe whether bird
activity or mobility was impacted. Diet was optimised for Cobb 500
birds and remained constant across all studies, as did stocking density
(28kg/m2 at 42 days old) and litter. All treatments had six replicates. In
study one and two, birds were given access to two metres of perch
space (one metre 15cm above the ground and one metre 30cm above
the ground) per pen (42 birds), and they were located in the corner of
the pens. In the third study, three metres of perch space was provided
per pen. These perches sat 10cm off the ground and ran down the length
of the pen between food and water, meaning birds had to cross them
to access either. In all three studies, behavioural observations were
made regularly, categorising all birds as either feeding, drinking,
active (standing/walking) or resting (perching/sitting). Separately, the
number of birds perching were counted to calculate perch rate.
Behaviour categories were analysed using repeated measures ANOVAs
with tukey post-hoc testing. Latency to lie (Berg & Sanotra 2003) was
assessed at 35 and 42 days old across all three studies to represent
mobility, and was analysed using a Kaplan-Meier survival analyses.
Perch use differed across the studies, with the third notably higher than
one and two (p=0.001) suggesting that increasing environmental
complexity increased motivation to perch in these birds. Despite this,
across all three studies the presence of perches did not influence
behaviour or activity levels. Likewise, the impact of perches on mobility
was minimal, with significance noted only at 42 days in study two
(p=0.013) wherein birds with access to perches had reduced mobility.
When compared, these studies suggest that perches as a form of
enrichment have negligible impact on bird activity levels and mobility,
even when motivation to use them is high.
ID: 997

EXAMINATION OF THE PLUMAGE CONDITION ON NON-BEAK-TRIMMED
LAYING HENS IN DIFFERENT KEEPING SYSTEMS

S.Szász*(1), G.Milisits(1), Z.Budai(2), A.Almási(3), P.Farkas(1), E.Garamvölgyi(1), J.Ujváriné(1), O.Kustosné(1), P.Horn(1), Z.Süto(1)

(1) Institute of Animal Science, Faculty of Agricultural and Environmental Sciences, Kaposvár
University, Kaposvár, Hungary, (2) Bábolna TETRA Ltd., Bábolna, Hungary, (3) Bábolna TETRA Ltd.,
Bábolna, Hungary

*Presenting author

**Corresponding author mail:** szasz.sandor@ke.hu (Sándor Szász)

The spread of non-cage laying hen housing systems and the more
forceful European refuse of beak trimming generate new problems in the
commercial egg production. The modern hybrid layers have vital
temperament, are susceptible for feather pecking and in more cases they
are expressly aggressive, which could cause permanent conflict situations
in the large group keeping systems. Therefore the aim of this study was
to examine the changes in the plumage condition of non-beak-trimmed
laying hens in cage and non-cage housing systems. The experiment was
carried out with altogether 1460 layers, representing two types: Rhode
Island Red (RIR) and Rhode Island White (RIW). The birds were kept in
traditional 3-tier cages* (630 cm2/hen), EU-compatible furnished cages
(756 cm2/hen), and floor system (slatted floor and deep litter, 1042
cm2/hen), in the same environmentally controlled laying house at the
Poultry Test Station of the Kaposvár University in Hungary. They were fed
ad libitum with commercial laying hen diet and drinking water was also
continuously available. The plumage condition of the hens was examined
at 33, 45 and 61 weeks of age. It was controlled at five different body
parts: neck, breast, wings, back and tail using a 4-grade scale, where
four points were given for the intact plumage and one point was given for
the strongly damaged plumage based on a reference photo series. By
summarizing the plumage points at the different body parts, a total
plumage point was calculated for each hen.

The total plumage point decreased in both examined types, but it was
significantly higher in the RIW than in the RIR hens at all of the
examined ages. The difference between the two types increased from
0.7 (18.3 and 19.0) to 1.7 (15.5 and 17.2) points between 33 and 61
weeks of age. The plumage was slightly damaged on the breast and
the back, meanwhile more damaged on the neck, tail and wings in
both types. From the examined keeping systems significantly better
results were obtained in the non-cage floor system than in the cage
systems.

Based on the results it was established that the RIW hens have more
calm temperament than the RIR hens and the non-caged floor system
seems to be more favourable for the plumage condition of the hens.

This research was supported by the '2018-1.3.1-VKE-2018-00042' project, funded from the Hungarian State.

*Permissions: MÁB 232.1/00412/0016/2011 and SOI/312766-7/2016 (KA-2044) Somogy
County Government Office.
AUTHOR INDEX

<table><tr><td>Abasht B.</td><td>ID:943</td></tr><tr><td>Abdollahi M.R.</td><td>ID:189</td></tr><tr><td>Abodi K.E.</td><td>ID:710</td></tr><tr><td>Abu Hafsa S.</td><td>ID:964</td></tr><tr><td>Adeola O.</td><td>ID:274</td></tr><tr><td>Ader P.</td><td>ID:887</td></tr><tr><td>Adjou K.</td><td>ID:1460</td></tr><tr><td>Aeschleman L.</td><td>ID:1104</td></tr><tr><td>Afzal A.</td><td>ID:1411 ID:1412</td></tr><tr><td>Agboka A.</td><td>ID:1234</td></tr><tr><td>Agboola A.</td><td>ID:1353</td></tr><tr><td>Aguihe P.</td><td>ID:1353</td></tr><tr><td>Ahmad S.</td><td>ID:667</td></tr><tr><td>Ahmad T.</td><td>ID:894</td></tr><tr><td>Aksit M.</td><td>ID:555</td></tr><tr><td>Albu A.</td><td>ID:1432</td></tr><tr><td>Allen B.</td><td>ID:1472</td></tr><tr><td>Almási A.</td><td>ID:997 ID:1262</td></tr><tr><td>Almeida Noleto R.</td><td>ID:950</td></tr><tr><td>Alves Da Costa M.</td><td>ID:931</td></tr><tr><td>Anchikov E.</td><td>ID:728</td></tr><tr><td>Andrés-Ramos I.</td><td>ID:993</td></tr><tr><td>Anwar A.</td><td>ID:897</td></tr><tr><td>Arne P.</td><td>ID:1460</td></tr><tr><td>Arnhold E.</td><td>ID:188 ID:1413</td></tr><tr><td>Arsenault R.</td><td>ID:1104</td></tr><tr><td>Asher L.</td><td>ID:1017</td></tr><tr><td>Ashraf S.</td><td>ID:1251</td></tr><tr><td>Atchiwassa S.</td><td>ID:710</td></tr><tr><td>Athanasiou L.</td><td>ID:723</td></tr><tr><td>Avital-Cohen N.</td><td>ID:1111 ID:1163</td></tr><tr><td>Awan N.</td><td>ID:752</td></tr><tr><td>Ayssiwede S.B.</td><td>ID:710</td></tr><tr><td>Azhaguraja M.</td><td>ID:110</td></tr><tr><td>Babatunde O.</td><td>ID:274</td></tr><tr><td>Baes C.F.</td><td>ID:945</td></tr><tr><td>Banaszak M.</td><td>ID:1282</td></tr><tr><td>Bang H.T.</td><td>ID:640 ID:642</td></tr><tr><td>Barbosa D.K.</td><td>ID:821</td></tr><tr><td>Barcellos Café M.</td><td>ID:931 ID:950</td></tr><tr><td>Barka E.</td><td>ID:1474</td></tr><tr><td>Barrios H.</td><td>ID:703</td></tr><tr><td>Barroeta Lajusticia A.C.</td><td>ID:993</td></tr><tr><td>Barros De Carvalho F.</td><td>ID:931</td></tr><tr><td>Bartman J.</td><td>ID:1111 ID:1163</td></tr><tr><td>Batallé M.</td><td>ID:703</td></tr><tr><td>Beck M.M.</td><td>ID:952</td></tr><tr><td>Beitia A.</td><td>ID:1450</td></tr><tr><td>Bello A.</td><td>ID:274</td></tr><tr><td>Bennewitz J.</td><td>ID:311</td></tr><tr><td>Bera C.</td><td>ID:1460</td></tr><tr><td>Berti-Sorbara J.</td><td>ID:901</td></tr><tr><td>Bhattacharya T.K.</td><td>ID:1266 ID:1273 ID:1277 ID:1285</td></tr><tr><td>Bhatti S.A.</td><td>ID:1251</td></tr><tr><td>Bhushan B.</td><td>ID:1277</td></tr></table>

<table><tr><td>Bibi F.</td><td>ID:1319</td></tr><tr><td>Biesek J.</td><td>ID:1282</td></tr><tr><td>Bigorowski B.</td><td>ID:1282</td></tr><tr><td>Bilgen G.</td><td>ID:555</td></tr><tr><td>Biondi M.</td><td>ID:703</td></tr><tr><td>Bogucka J.</td><td>ID:1282</td></tr><tr><td>Bonos E.</td><td>ID:1474</td></tr><tr><td>Bonou M.C.</td><td>ID:710</td></tr><tr><td>Borda-Molina D.</td><td>ID:311</td></tr><tr><td>Borges S.</td><td>ID:481</td></tr><tr><td>Budai Z.</td><td>ID:997</td></tr><tr><td>Buitrago D.</td><td>ID:408</td></tr><tr><td>Burbarelli M.F.</td><td>ID:821</td></tr><tr><td>Burin Á.M.</td><td>ID:1220</td></tr><tr><td>Burin Junior A.M.</td><td>ID:1005</td></tr><tr><td>Butt S.L.</td><td>ID:1411</td></tr><tr><td>Cachoni Pizzolante C.</td><td>ID:315</td></tr><tr><td>Cafe M.</td><td>ID:988 ID:1413</td></tr><tr><td>Caldas J.</td><td>ID:1450</td></tr><tr><td>Calderano A.</td><td>ID:481</td></tr><tr><td>Camarinha-Silva A.</td><td>ID:311</td></tr><tr><td>Campos P.</td><td>ID:481</td></tr><tr><td>Carvalho F.</td><td>ID:950</td></tr><tr><td>Carvalho G.</td><td>ID:988</td></tr><tr><td>Carvalho L.V.</td><td>ID:999</td></tr><tr><td>Carvalho Rodrigues J.</td><td>ID:230</td></tr><tr><td>Castañeda C.D.</td><td>ID:952</td></tr><tr><td>Castellanos L.</td><td>ID:408 ID:415</td></tr><tr><td>Castilho V.A.</td><td>ID:821</td></tr><tr><td>Castro F.</td><td>ID:570</td></tr><tr><td>Chandan</td><td>ID:1191</td></tr><tr><td>Chatterjee R.N.</td><td>ID:635 ID:1183 ID:1266 ID:1273 ID:1277 ID:1285</td></tr><tr><td>Chen H.</td><td>ID:1179</td></tr><tr><td>Chen Y.</td><td>ID:581</td></tr><tr><td>Chico M.</td><td>ID:408</td></tr><tr><td>Chimienti C.</td><td>ID:1460</td></tr><tr><td>Choi P.</td><td>ID:217</td></tr><tr><td>Chrysostome C.</td><td>ID:1234</td></tr><tr><td>Chukkala S.</td><td>ID:1285</td></tr><tr><td>Chun J.L.</td><td>ID:640 ID:642</td></tr><tr><td>Clark C.</td><td>ID:897</td></tr><tr><td>Collin A.</td><td>ID:1234</td></tr><tr><td>Combes-Soia L.</td><td>ID:288</td></tr><tr><td>Coon C.</td><td>ID:1450</td></tr><tr><td>Cordeiro D.</td><td>ID:188 ID:1413</td></tr><tr><td>Cordova H.</td><td>ID:408 ID:415</td></tr><tr><td>Costa M.A.</td><td>ID:988</td></tr><tr><td>Craven M.J.</td><td>ID:1020</td></tr><tr><td>Crowley T.M.</td><td>ID:675 ID:1020</td></tr><tr><td>Csitári G.</td><td>ID:805 ID:813</td></tr><tr><td>Cui Y.</td><td>ID:855</td></tr><tr><td>Da Silva Oliveira G.</td><td>ID:230</td></tr><tr><td>Dai D.</td><td>ID:792</td></tr><tr><td>Dalton H.A.</td><td>ID:945</td></tr></table>
<table><tr><td>Dange M.</td><td>ID:1266</td></tr><tr><td>Dankowiakowska A.</td><td>ID:1282</td></tr><tr><td>Dash S.</td><td>ID:1082</td></tr><tr><td>De Franceschi M.</td><td>ID:703</td></tr><tr><td>De Oliveira H.</td><td>ID:188 ID:1413</td></tr><tr><td>De Laet M.</td><td>ID:214</td></tr><tr><td>De Souza Eberhart</td><td>ID:821</td></tr><tr><td>De Souza Vieira M.</td><td>ID:704</td></tr><tr><td>Dean L.</td><td>ID:999</td></tr><tr><td>Decuypere E.</td><td>ID:1234</td></tr><tr><td>Deeve Sabastein G.</td><td>ID:739</td></tr><tr><td>Dekkers J.C.</td><td>ID:943</td></tr><tr><td>Delago J.</td><td>ID:1084</td></tr><tr><td>Dersjant-Li Y.</td><td>ID:274</td></tr><tr><td>Devillard E.</td><td>ID:217 ID:675</td></tr><tr><td>Dias F.</td><td>ID:214</td></tr><tr><td>Ding C.</td><td>ID:1411 ID:1412</td></tr><tr><td>Dishon L.</td><td>ID:1111 ID:1163</td></tr><tr><td>Divya D.</td><td>ID:1273</td></tr><tr><td>Djitie Kouatcho F.</td><td>ID:1432</td></tr><tr><td>Donkó T.</td><td>ID:1262</td></tr><tr><td>Dos Santos V.M.</td><td>ID:230</td></tr><tr><td>Du H.</td><td>ID:614</td></tr><tr><td>Duarte M.</td><td>ID:481</td></tr><tr><td>Dublecz K.</td><td>ID:805 ID:813 ID:1001 ID:1004</td></tr><tr><td>Đukic Stojcic M.</td><td>ID:813</td></tr><tr><td>Dutra F.M.</td><td>ID:821</td></tr><tr><td>Dzungwe J.T.</td><td>ID:513 ID:739</td></tr><tr><td>Egahi J.</td><td>ID:513</td></tr><tr><td>Emamgholi Begli H.</td><td>ID:945</td></tr><tr><td>Etheve S.</td><td>ID:1242</td></tr><tr><td>Ezagal J.</td><td>ID:288</td></tr><tr><td>Farkas P.T.</td><td>ID:997</td></tr><tr><td>Farkas T.P.</td><td>ID:1262</td></tr><tr><td>Farkas V.</td><td>ID:805 ID:813</td></tr><tr><td>Farooq U.</td><td>ID:894</td></tr><tr><td>Fawzy M.</td><td>ID:592</td></tr><tr><td>Fébel H.</td><td>ID:1004</td></tr><tr><td>Feng J.</td><td>ID:794</td></tr><tr><td>Ferket P.</td><td>ID:999</td></tr><tr><td>Fernandes filho I.</td><td>ID:188</td></tr><tr><td>Fernando Machado Me J.</td><td>ID:1005</td></tr><tr><td>Fernando Sens R.</td><td>ID:1242</td></tr><tr><td>Ferreira Pires M.</td><td>ID:950</td></tr><tr><td>Ferreira Pires S.</td><td>ID:950</td></tr><tr><td>Fialho De Sousa R.</td><td>ID:931</td></tr><tr><td>Fontes M.</td><td>ID:481</td></tr><tr><td>Fotou K.</td><td>ID:1474</td></tr><tr><td>Fu Y.</td><td>ID:790 ID:791</td></tr><tr><td>Fuller A.</td><td>ID:570</td></tr><tr><td>Galliard N.</td><td>ID:1460</td></tr><tr><td>Galovic O.</td><td>ID:121</td></tr><tr><td>Garamvölgyi E.</td><td>ID:997 ID:1262</td></tr><tr><td>Garcia R.G.</td><td>ID:821</td></tr><tr><td>Gautron J.</td><td>ID:288</td></tr><tr><td>Gbeassor M.</td><td>ID:1234</td></tr><tr><td>Gebhardt-Henrich S.G.</td><td>ID:1017</td></tr><tr><td>Geier R.</td><td>ID:1084</td></tr><tr><td>Gharib-Naseri K.</td><td>ID:675</td></tr><tr><td>Ghayas A.</td><td>ID:667</td></tr></table>

<table><tr><td>Giorgi USTUROI M.</td><td>ID:1432</td></tr><tr><td>Glendinning L.</td><td>ID:127</td></tr><tr><td>Gogoi S.</td><td>ID:991</td></tr><tr><td>Gomes Guimarães I.</td><td>ID:931</td></tr><tr><td>Gomez Y.</td><td>ID:987</td></tr><tr><td>Gómez Y.</td><td>ID:1017</td></tr><tr><td>González J.</td><td>ID:481</td></tr><tr><td>Groves P.</td><td>ID:280</td></tr><tr><td>Guerrero-Bosagna C.</td><td>ID:1017</td></tr><tr><td>Guiscafre F.</td><td>ID:524</td></tr><tr><td>Gunn H.H.</td><td>ID:682</td></tr><tr><td>Gwaza S.</td><td>ID:513</td></tr><tr><td>Haeinger V.</td><td>ID:704</td></tr><tr><td>Hai L.</td><td>ID:1158</td></tr><tr><td>Han Y. ID:315 ID:381</td><td>ID:792 ID:882</td></tr><tr><td>Hanzek D.</td><td>ID:121</td></tr><tr><td>Hao E.</td><td>ID:1179</td></tr><tr><td>Harlander-Matausche A.</td><td>ID:945</td></tr><tr><td>Harrington D.</td><td>ID:214</td></tr><tr><td>Hassan A.</td><td>ID:964</td></tr><tr><td>Hassanzadeh M.</td><td>ID:555</td></tr><tr><td>Heffron J.</td><td>ID:217</td></tr><tr><td>Heidysz M.</td><td>ID:1418</td></tr><tr><td>Hejdysz M.</td><td>ID:827</td></tr><tr><td>Helva B.</td><td>ID:555</td></tr><tr><td>Henning J.</td><td>ID:659</td></tr><tr><td>Hilton K.</td><td>ID:1450</td></tr><tr><td>Hincke M.</td><td>ID:288</td></tr><tr><td>Hirunpatrawong W.</td><td>ID:1084</td></tr><tr><td>Holden D.</td><td>ID:897</td></tr><tr><td>Horn P.</td><td>ID:997 ID:1262</td></tr><tr><td>Hoyos S.</td><td>ID:408</td></tr><tr><td>Hu B.</td><td>ID:1139</td></tr><tr><td>Hu Y.</td><td>ID:882</td></tr><tr><td>Huang R.</td><td>ID:1179</td></tr><tr><td>Hussain J.</td><td>ID:627 ID:667</td></tr><tr><td>Husvéth F.</td><td>ID:930</td></tr><tr><td>Iqbal H.H.</td><td>ID:1001 ID:1004</td></tr><tr><td>Iqbal J.</td><td>ID:1251</td></tr><tr><td>Iyayi E.</td><td>ID:894</td></tr><tr><td>Izzetoglu G.T.</td><td>ID:1353</td></tr><tr><td>Jakkula R.</td><td>ID:555</td></tr><tr><td>Jankowski J.</td><td>ID:1253</td></tr><tr><td>Jansen Van Rensburg C.</td><td>ID:784</td></tr><tr><td>Jardim filho R.</td><td>ID:1413</td></tr><tr><td>Jeong J.Y.</td><td>ID:640 ID:642</td></tr><tr><td>Ji S.Y.</td><td>ID:640 ID:642</td></tr><tr><td>Jia Y.</td><td>ID:581</td></tr><tr><td>Jiang X.</td><td>ID:614</td></tr><tr><td>Joseph E.</td><td>ID:739</td></tr><tr><td>Jozefiak A.</td><td>ID:229</td></tr><tr><td>Józefiak D.</td><td>ID:229</td></tr><tr><td>Juskiewicz J.</td><td>ID:229</td></tr><tr><td>Kaczmarek S.</td><td>ID:827</td></tr><tr><td>Kaiser De Lima-Kren F.</td><td>ID:381</td></tr><tr><td>Kannan A.</td><td>ID:635 ID:637 ID:1092</td></tr><tr><td>Karamoutsios A.</td><td>ID:1474</td></tr><tr><td>Kashif Yameen M.</td><td>ID:627</td></tr><tr><td>Kazemifard M.</td><td>ID:1444</td></tr><tr><td>Keerqin C.</td><td>ID:675</td></tr></table>
<table><tr><td>Kemp B.</td><td>ID:1234</td></tr><tr><td>Khan D.</td><td>ID:827</td></tr><tr><td>Kheravii S.K.</td><td>ID:675</td></tr><tr><td>Khucharoentaworn S.</td><td>ID:1307</td></tr><tr><td>Kiess A.S.</td><td>ID:952</td></tr><tr><td>Kim B.</td><td>ID:640 ID:642</td></tr><tr><td>Kim C.</td><td>ID:342</td></tr><tr><td>Kim K.H.</td><td>ID:640 ID:642</td></tr><tr><td>Kim M.J.</td><td>ID:640 ID:642</td></tr><tr><td>Kim W.</td><td>ID:570</td></tr><tr><td>Kolluri G.</td><td>ID:991</td></tr><tr><td>Komarov A.</td><td>ID:244</td></tr><tr><td>Komiyama C.M.</td><td>ID:821</td></tr><tr><td>Koutoulis K.</td><td>ID:723</td></tr><tr><td>Koutsianos D.</td><td>ID:723</td></tr><tr><td>Kralik G.</td><td>ID:121</td></tr><tr><td>Kralik I.</td><td>ID:172</td></tr><tr><td>Kralik Z.</td><td>ID:121</td></tr><tr><td>Kramer L.</td><td>ID:943</td></tr><tr><td>Kreuz B.</td><td>ID:481</td></tr><tr><td>Kumar P.</td><td>ID:1266 ID:1277</td></tr><tr><td>Kustosné O.P.</td><td>ID:997 ID:1262</td></tr><tr><td>Labas V.</td><td>ID:288</td></tr><tr><td>Lake J.</td><td>ID:943</td></tr><tr><td>Latha Preethi A.</td><td>ID:1266</td></tr><tr><td>Le Roy N.</td><td>ID:288</td></tr><tr><td>Lemme A.</td><td>ID:438</td></tr><tr><td>Lemos Budiño F.E.</td><td>ID:315</td></tr><tr><td>Lemos De Moraes M.</td><td>ID:704</td></tr><tr><td>Lesli L.</td><td>ID:1191</td></tr><tr><td>Li H.</td><td>ID:1139</td></tr><tr><td>Li J.</td><td>ID:1299</td></tr><tr><td>Li Q.</td><td>ID:614</td></tr><tr><td>Li X.</td><td>ID:581</td></tr><tr><td>Liu H.</td><td>ID:901</td></tr><tr><td>Liu X.</td><td>ID:1299</td></tr><tr><td>Livingston K.</td><td>ID:999</td></tr><tr><td>Livingston M.</td><td>ID:999</td></tr><tr><td>Lopes M.</td><td>ID:481</td></tr><tr><td>Lukashenko V.</td><td>ID:244</td></tr><tr><td>Lv X.</td><td>ID:581</td></tr><tr><td>Maake M.S.</td><td>ID:543</td></tr><tr><td>Mabelebele M.</td><td>ID:543</td></tr><tr><td>Mackei M.</td><td>ID:1001 ID:1004</td></tr><tr><td>Maharjan P.</td><td>ID:1450</td></tr><tr><td>Mahmud A.</td><td>ID:627 ID:667</td></tr><tr><td>Maiorka A.</td><td>ID:1242</td></tr><tr><td>Malheiros R.</td><td>ID:999</td></tr><tr><td>Männer K.</td><td>ID:887</td></tr><tr><td>Mannickam K.</td><td>ID:991</td></tr><tr><td>Marappan G.</td><td>ID:991</td></tr><tr><td>Marquis V.</td><td>ID:322</td></tr><tr><td>Martinez D.</td><td>ID:1450</td></tr><tr><td>Martinez J.</td><td>ID:408</td></tr><tr><td>Martinez-Vallespin B.</td><td>ID:887</td></tr><tr><td>Martins J.</td><td>ID:988</td></tr><tr><td>Mascarenhas A.</td><td>ID:188</td></tr><tr><td>Mátis G.</td><td>ID:1001 ID:1004</td></tr><tr><td>Matta Y.</td><td>ID:408</td></tr><tr><td>Mcdaniel C.D.</td><td>ID:952</td></tr></table>

<table><tr><td>Mehmood S.</td><td>ID:667</td></tr><tr><td>Mello H.</td><td>ID:188</td></tr><tr><td>Melo J.</td><td>ID:703</td></tr><tr><td>Meng C.</td><td>ID:1411 ID:1412</td></tr><tr><td>Mens A.</td><td>ID:524</td></tr><tr><td>Menten J.F.</td><td>ID:1220</td></tr><tr><td>Michelin Groff-Uray P.</td><td>ID:381</td></tr><tr><td>Mikulski D.</td><td>ID:229</td></tr><tr><td>Milisits G.</td><td>ID:997 ID:1262</td></tr><tr><td>Mishra S.</td><td>ID:1082</td></tr><tr><td>Missohou A.O.</td><td>ID:710</td></tr><tr><td>Mogyca Leandro N.</td><td>ID:950</td></tr><tr><td>Molnar A.</td><td>ID:805</td></tr><tr><td>Molnár A.</td><td>ID:813 ID:1001 ID:1004</td></tr><tr><td>Moongkundar C.</td><td>ID:1307</td></tr><tr><td>Moraes Cruvinel J.</td><td>ID:381</td></tr><tr><td>Moraes J.E.</td><td>ID:315</td></tr><tr><td>Moraes Rezende P.</td><td>ID:931</td></tr><tr><td>Morgan N.K.</td><td>ID:207</td></tr><tr><td>Mossialos D.</td><td>ID:723</td></tr><tr><td>Muir W.</td><td>ID:280</td></tr><tr><td>Mukhtar N.</td><td>ID:752 ID:894</td></tr><tr><td>Muralidhar M.</td><td>ID:1285</td></tr><tr><td>Nagy S.</td><td>ID:1001</td></tr><tr><td>Naim A.</td><td>ID:1082 ID:1082</td></tr><tr><td>Narvaez M.</td><td>ID:1401</td></tr><tr><td>Nascimento L.E.</td><td>ID:821</td></tr><tr><td>Nascimento-Filho M.A.</td><td>ID:1005 ID:1220</td></tr><tr><td>Naveed S.</td><td>ID:1319</td></tr><tr><td>Nayak D.</td><td>ID:1082</td></tr><tr><td>Neerasa G.S.</td><td>ID:1277</td></tr><tr><td>Nelson A.</td><td>ID:217</td></tr><tr><td>Neogrády Z.</td><td>ID:1001 ID:1004</td></tr><tr><td>Netrabukkana P.</td><td>ID:1307</td></tr><tr><td>Nguyen T.T.</td><td>ID:207</td></tr><tr><td>Nie C.</td><td>ID:581</td></tr><tr><td>Nikonov I.</td><td>ID:728</td></tr><tr><td>Ning Z.</td><td>ID:581</td></tr><tr><td>Nunes R.</td><td>ID:481</td></tr><tr><td>Nunes Souto C.</td><td>ID:931</td></tr><tr><td>Odu O.</td><td>ID:1353</td></tr><tr><td>Ofongo R.T.</td><td>ID:1472</td></tr><tr><td>Ogbonna V.C.</td><td>ID:682</td></tr><tr><td>Omidiwura R.</td><td>ID:1353</td></tr><tr><td>Omprakash A.V.</td><td>ID:574 ID:1018</td></tr><tr><td>Onagbesan O.M.</td><td>ID:1234</td></tr><tr><td>Orbán A.</td><td>ID:1262</td></tr><tr><td>Ortiz A.</td><td>ID:408</td></tr><tr><td>Ospina-Rojas I.</td><td>ID:408</td></tr><tr><td>Oviedo-Rondón E.</td><td>ID:408 ID:415</td></tr><tr><td>Ovseychik E.</td><td>ID:244</td></tr><tr><td>Owen O.J.</td><td>ID:682</td></tr><tr><td>Ozkan S.</td><td>ID:555</td></tr><tr><td>Pal L.</td><td>ID:805</td></tr><tr><td>Pál L.</td><td>ID:813</td></tr><tr><td>Pallen M.J.</td><td>ID:127</td></tr><tr><td>Pandian C.</td><td>ID:1018</td></tr><tr><td>Pasha T.N.</td><td>ID:1319</td></tr><tr><td>Paswan C.</td><td>ID:1266 ID:1277</td></tr><tr><td>Patterson R.</td><td>ID:229</td></tr></table>
Paul S. ID:1092
Paul S.S. ID:635 ID:637
ID:1183 ID:1253

Paulpandi T.G. ID:574

Peñuela L. ID:408

Pereira Carvalho D. ID:950

Pereira R.T. ID:1220

Perera W.N. ID:189

Pérez A.M. ID:322

Peric L. ID:813

Perry F. ID:1104

Pertille F. ID:1017

Perz K. ID:1418

Phibbs D. ID:280

Pietramale R. ID:821

Pinto S. ID:703

Prakash B. ID:635 ID:637
ID:1092 ID:1183

Prasad A.R. ID:1277

Prasad M. ID:1092

Preveraud D. ID:217

Prosdócimo F. ID:703

Przybulinski B.B. ID:821

Pym R. ID:659

Qaisrani S.N. ID:1319

Qi G. ID:790 ID:791

ID:792 ID:794

ID:855

Qiu M. ID:614

Qu L. ID:581

Quinteros J.A. ID:897

Rabelo Ribeiro A. ID:931

Radu-Rusu R.M. ID:1432

Rajkumar . ID:1191

Raju M.V. ID:635 ID:637
ID:1092 ID:1183
ID:1191

Rama Rao S.V. ID:1092 ID:1183

ID:1191 ID:1253

Ramos A.J. ID:322

Rao S. ID:1450

Rao S.V. ID:635 ID:637

Ravindran V. ID:189

Ray B.C. ID:2222

Reddy C. ID:1092

Rehberger T. ID:1084

Rehman A. ID:667

Rehman Z.U. ID:1411 ID:1412

Ren S. ID:1411 ID:1412

Reynecke A. ID:784

Rezaei M. ID:1444

Rezaei S. ID:1017

Rhayat L. ID:217 ID:675

Riahi I. ID:322

Rice J. ID:217

Rios L. ID:1401

Roberts J.R. ID:207

Rodehutscord M. ID:311

Rodenburg T.B. ID:987

Rodríguez-Navaroo A. ID:288

Rodríguez-Sánchez R. ID:993

Rogiewicz A. ID:229
Romero L.R. ID:1220
Romlamduan N. ID:1307
Roth C. ID:311
Rouhparvar H. ID:1444
Rozenboim I. ID:1111 ID:1163
Rudranath ID:1191
Rutkowski A. ID:1418
Sahar N.U. ID:1251
Saima S. ID:627
Salvador E. ID:1401
San Martin V. ID:408
Sanda K. ID:1234
Sanders E. ID:999
Santin E. ID:704
Santos Batistioli J. ID:381
Santos De Oliveira A.B. ID:1005
ID:1220

Sarfraz M.A. ID:1319

Sartori J.R. ID:381

Saxena V.K. ID:991

Scharch C. ID:438

Schmid M. ID:311

Schmidt Bassi L. ID:1242

Scott P.C. ID:897

Scott T. ID:897

Selaledi L.A. ID:543

Selvan S.T. ID:1018

Serpa ID:821

Sgavioli S. ID:988

Shadpour S. ID:945

Shahid F. ID:1251

Shanmuga Sundaram A. ID:574

Shi S. ID:882

Shoaib M. ID:1251

Sidelmann Brinch K. ID:217

Sijtsma L. ID:524

Sithole T. ID:980

Skoufos I. ID:1474

Skoufos S. ID:1474

Slominski B. ID:229

Smith A. ID:1084

Sosa N. ID:703

Sousa F.M. ID:987

Souza Dos Santos T. ID:381

Stapane L. ID:288

Stefanello C. ID:704

Stewart R.D. ID:127

Steyl D. ID:784

Stratmann A. ID:1017

Stringhini J.H. ID:188 ID:931
ID:950 ID:988
ID:1413

Subbiah E.V. ID:574 ID:1018

Such N. ID:805 ID:813

Suckeveris D. ID:1005 ID:1220

Suesuttajit N. ID:1450

Sun Y. ID:1411 ID:1412

Sundaresan A. ID:1018

Susana Mogica Leandro N. ID:931

Sushma G. ID:1277
<table><tr><td>Süto Z.</td><td>ID:997 ID:1262</td></tr><tr><td>Svetlana S.</td><td>ID:1297</td></tr><tr><td>Swatson H.</td><td>ID:980</td></tr><tr><td>Swick R.A.</td><td>ID:207</td></tr><tr><td>Szász S.</td><td>ID:997 ID:1262</td></tr><tr><td>Taechavasonyoo A.</td><td>ID:1084</td></tr><tr><td>Tatiane Pereira R.</td><td>ID:1005</td></tr><tr><td>Taylor P.S.</td><td>ID:1020</td></tr><tr><td>Teixeira L.</td><td>ID:1242</td></tr><tr><td>Teng P.</td><td>ID:570</td></tr><tr><td>Thapelo K.</td><td>ID:980</td></tr><tr><td>Theja Sri J.</td><td>ID:1277</td></tr><tr><td>Thiery P.</td><td>ID:675</td></tr><tr><td>Toghyani M.</td><td>ID:207</td></tr><tr><td>Toledo Santos C.</td><td>ID:381</td></tr><tr><td>Tompkins Y.</td><td>ID:570</td></tr><tr><td>Tona K.</td><td>ID:1234</td></tr><tr><td>Toomer O.</td><td>ID:999</td></tr><tr><td>Torrey S.</td><td>ID:945</td></tr><tr><td>Toscano M.J.</td><td>ID:987 ID:1017</td></tr><tr><td>Tulpan D.</td><td>ID:945</td></tr><tr><td>Tyagi J.S.</td><td>ID:991</td></tr><tr><td>Tzora A.</td><td>ID:1474</td></tr><tr><td>Ujváriné J.</td><td>ID:997 ID:1262</td></tr><tr><td>Uvertse E.N.</td><td>ID:682</td></tr><tr><td>Valentim J.K.</td><td>ID:821</td></tr><tr><td>Van Immerseel F.</td><td>ID:217</td></tr><tr><td>Van Krimpen M.</td><td>ID:524</td></tr><tr><td>Van Marle-Koster E.</td><td>ID:784</td></tr><tr><td>Vang E.</td><td>ID:1084</td></tr><tr><td>Vanni Jacob D.</td><td>ID:950</td></tr><tr><td>Verge-Mèrida G.</td><td>ID:993</td></tr><tr><td>Vieira De Andrade T.</td><td>ID:931</td></tr><tr><td>Vignoni E.</td><td>ID:703</td></tr><tr><td>Vinoo R.</td><td>ID:1285</td></tr><tr><td>Vögeli S.</td><td>ID:1017</td></tr><tr><td>Voidarou C.</td><td>ID:1474</td></tr><tr><td>Völkl B.</td><td>ID:1017</td></tr><tr><td>Vollmar S.</td><td>ID:311</td></tr><tr><td>Von Zuben Augusto K.</td><td>ID:315 ID:381</td></tr><tr><td>Vu T.</td><td>ID:999</td></tr><tr><td>Wagner L.</td><td>ID:805</td></tr><tr><td>Wágner L.</td><td>ID:813</td></tr><tr><td>Wall B.</td><td>ID:999</td></tr><tr><td>Walsh M.</td><td>ID:901</td></tr><tr><td>Walugembe M.</td><td>ID:943</td></tr><tr><td>Wamsley K.G.</td><td>ID:952</td></tr></table>

<table><tr><td>Wang J.</td><td>ID:790 ID:791</td></tr><tr><td>Wang J.</td><td>ID:792 ID:794 ID:855</td></tr><tr><td>Wang L.</td><td>ID:581</td></tr><tr><td>Wang W.</td><td>ID:791</td></tr><tr><td>Wang X.</td><td>ID:1158</td></tr><tr><td>Wang Y.</td><td>ID:614</td></tr><tr><td>Watson K.A.</td><td>ID:127</td></tr><tr><td>Watson M.</td><td>ID:127</td></tr><tr><td>Wei H.</td><td>ID:1139</td></tr><tr><td>Weil J.</td><td>ID:1450</td></tr><tr><td>Wester T.J.</td><td>ID:189</td></tr><tr><td>Wilson J.W.</td><td>ID:1242</td></tr><tr><td>Wilson T.B.</td><td>ID:897</td></tr><tr><td>Witjes V.L.</td><td>ID:987</td></tr><tr><td>Wolfende R.</td><td>ID:217</td></tr><tr><td>Wood B.J.</td><td>ID:945</td></tr><tr><td>Wu J.</td><td>ID:901</td></tr><tr><td>Wu S.</td><td>ID:675 ID:790 ID:791 ID:792 ID:794 ID:855</td></tr><tr><td>Wu Y.</td><td>ID:792 ID:882</td></tr><tr><td>Yadav S.</td><td>ID:570</td></tr><tr><td>Yalcin S.</td><td>ID:555</td></tr><tr><td>Yang C.</td><td>ID:614</td></tr><tr><td>Yang W.</td><td>ID:581 ID:1299</td></tr><tr><td>Yanquen J.</td><td>ID:408</td></tr><tr><td>Yaseen G.</td><td>ID:1319</td></tr><tr><td>Yilmaz M.C</td><td>ID:555</td></tr><tr><td>Yin H.</td><td>ID:614</td></tr><tr><td>Yu C.</td><td>ID:614</td></tr><tr><td>Yutto Oura C.</td><td>ID:381</td></tr><tr><td>Zaefarian F.</td><td>ID:189</td></tr><tr><td>Zaguri S.</td><td>ID:1111 ID:1163</td></tr><tr><td>Zaid M.</td><td>ID:930</td></tr><tr><td>Zasorin A.</td><td>ID:728</td></tr><tr><td>Zdunczyk Z.</td><td>ID:229</td></tr><tr><td>Zentek J.</td><td>ID:887</td></tr><tr><td>Zhang H.</td><td>ID:790 ID:791 ID:792 ID:794 ID:855</td></tr><tr><td>Zhang J.</td><td>ID:581</td></tr><tr><td>Zhang L.</td><td>ID:952</td></tr><tr><td>Zhang Q.</td><td>ID:901</td></tr><tr><td>Zhang X.</td><td>ID:1139 ID:1299</td></tr><tr><td>Zhang Z.</td><td>ID:614 ID:675</td></tr></table>
List of our sponsors

Diamond

ADISSEO
A Biocenter Company

<figure><img src="image_5.png" /></figure>

Together, beyond animal health

Platinum

EVONIK
Leading Beyond Chemistry

Gold

<figure><img src="image_6.png" /></figure>

DSM

BRIGHT SCIENCE, BRIGHTER LIVING

Elanco

<figure><img src="image_7.png" /></figure>

MSD

Animal Health

<figure><img src="image_8.png" /></figure>

NOOVISTAGO

<figure><img src="image_9.png" /></figure>

NOVUS

Silver

iMarel

Bronze

agrimprove
we change lives

CCPA
GROUP

<figure><img src="image_10.png" /></figure>

CHR HANSEN

Improving food & health

<figure><img src="image_11.png" /></figure>

BIO

Delacon

performing nature

iff

id4feed
for a better world

<figure><img src="image_12.png" /></figure>

innovad.

CREATE TRUST

<figure><img src="image_13.png" /></figure>

kersia

INDUSTRIAL + FOOD SAFE WORLD

<figure><img src="image_14.png" /></figure>

trouw nutrition

a NUTRACO company

<figure><img src="image_15.png" /></figure>

ZINPRO

Other partners

<figure><img src="image_16.png" /></figure>

AeroCollect®

SEPPIC

<figure><img src="image_17.png" /></figure>

Aviagen

<figure><img src="image_18.png" /></figure>

HENDRIX

<figure><img src="image_19.png" /></figure>

GENETICS

<figure><img src="image_20.png" /></figure>

HUVEPHARMA

L. RETTENMANN & SCHÜE

IRG

<figure><img src="image_21.png" /></figure>

Jefo

Lifecare.com

Natur

<figure><img src="image_22.png" /></figure>

Add

The Specialty Feed Ingredient Platform

<figure><img src="image_23.png" /></figure>

Perstorp

<figure><img src="image_24.png" /></figure>

Phileo

by Lesaffre

wisium

NUTRITION & BEYOND

Media partners

<figure><img src="image_25.png" /></figure>

<figure><img src="image_26.png" /></figure>

middle east agrifood publishers

aviNews.com

<figure><img src="image_27.png" /></figure>

WATTPoultry

International