Performance, health and tissue weights of broiler chickens fed graded levels of hairless hulled yellow and brown canary seed (Phalaris canariensis L.

Performance, health and tissue weights of broiler chickens fed graded levels of hairless hulled yellow and brown canary seed (Phalaris canariensis L.) Henry Classen 1, Misaki Cho 1, Pierre Hucl 2, Susantha Gomis 3, and Carol Ann Patterson 4 1 University of Saskatchewan, Animal and Poultry Science, 51 Campus Drive, Saskatoon, Saskatchewan, Canada S7N 5A8; 2 University of Saskatchewan, Crop Development Centre, Saskatoon, Saskatchewan, Canada S7N 5A8; 3 University of Saskatchewan, Veterinary Pathology Saskatchewan, Saskatchewan, Canada S7N 5B4; and 4 The Pathfinders Research & Management Ltd, Saskatoon, Saskatchewan, Canada S7N 0S5. Received 8 April 2014, accepted 14 August 2014. Published on the web 21 August 2014. Classen, H., Cho, M., Hucl, P., Gomis, S. and Patterson, C. A. 2014. Performance, health and tissue weights of broiler chickens fed graded levels of hairless hulled yellow and brown canary seed (Phalaris canariensis L.). Can. J. Anim. Sci. 94: 669678. Research examined the effects of dietary levels (0, 15, 30 and 45%) of hulled yellow (C05041) and brown (CDC Maria) canary seed on the performance and health of broiler chickens. Eight (six birds each) and five (four birds each) replications per treatment were used from 0 to 21 and from 22 to 35 d of age, respectively. Growth rate and feed intake were affected in a quadratic manner by canary seed level from 0 to 21 d with the highest growth achieved by 15 and 30% treatments, but were not affected from 22 to 35 d. Feed to gain ratio decreased linearly with increasing canary seed for both 0- to 21-d and 22- to 35-d time periods. Mortality was not affected by level of canary seed, but more birds fed the C05041 cultivar died as a result of yolk sac infection and infectious causes than did birds fed CDC Maria. Level of canary seed affected jejunum, kidney and bursa of Fabricius proportional weights in a quadratic manner with no differences between the 0 and 45% treatments. No interactions were found between dietary level and cultivar of canary seed. Treatment did not affect gross necropsy at trial end or histopathology of key organs. Yellow and brown hairless canary seed has merit and is safe for poultry feeding. Key words: Canary seed, poultry, seed colour, feed Classen, H., Cho, M., Hucl, P., Gomis, S. et Patterson, C. A. 2014. Performance, sante et poids des tissus des poulets à griller nourris des niveaux croissants de graines a` canaris (Phalaris canariensis L.) jaunes et brunes sans poils et nues Can. J. Anim. Sci. 94: 669678. La recherche avait comme objectif l examen des effets des niveaux alimentaires (0, 15, 30 et 45 %) des graines a` canaris nues jaunes (C05041) et brunes (CDC Maria) sur la performance et la santé des poulets a` griller. Huit re plicats (6 poulets chacun) et cinq re plicats (4 poulets chacun) par traitement ont e té utilise s (0 a` 21 et 22 a` 35 jours d aˆge, respectivement). Il y a eu un effet quadratique du niveau de graines a` canaris sur le taux de croissance et la prise alimentaire des jours 0 à 21, avec la plus grande croissance observée avec les traitements de 15 et 30 %. Il n y a eu aucun effet des jours 22 à 35. L indice de consommation a diminue de fac on line aire avec une augmentation des graines a` canaris pour les deux pe riodes de temps (0 a` 21 et 22 a` 35 jours). Il n y a pas eu d effet du niveau de graines a` canaris sur la mortalite, mais plus de poulets nourris au cultivar C05041 sont de ce de s a` la suite d infection du sac vitellin et autres causes infectieuses que les poulets nourris au cultivar CDC Maria. Le niveau de graines a` canaris avait un effet quadratique sur les poids proportionnels du je junum, du rein et de la bourse de Fabricius; il n y avait aucune diffe rence entre les traitements 0 et 45 %. Aucune interaction n a été observe e entre les niveaux alimentaires et du cultivar de la graine a` canaris. Le traitement n a eu aucun effet sur l autopsie macroscopique pratique e àla fin de l expe rience ni sur l histopathologie d organes-cle s. Les graines a` canaris jaunes et brunes sans poils sont be ne fiques et se curitaires pour l alimentation des poulets. Mots clés: Graines à canaris, poulets, coloration de la graine, alimentation Annual canary seed (Phalaris canariensis L.) is a specialty crop that is mostly grown and produced in Canada (Food and Agriculture Organization of the United Nations 2011). Saskatchewan produced 101 900 t of canary seed in 2010 and is responsible for more than 70% of the value of world canaryseed exports (Food and Agriculture Organization of the United Nations 2011; Government of Saskatchewan 2011). Both hairy and hairless hulled canary seed are grown in Saskatchewan. The hairy canary seed has hulls that are covered in silica fibre, which can cause skin irritation during handling and may also cause esophageal cancer (O Neill et al. 1980). Hence, hairless canary seed varieties were developed by the Crop Development Centre (University of Saskatchewan) to eliminate the potential health problems related to hairy canary seed varieties (Hucl et al. 2001). Three hairless varieties are currently registered in Canada (CDC Maria, CDC Togo and CDC Bastia) and these are all characterized by brown seed coats. A yellow seed coloured hairless variety has also been developed (C05041) with the expectation that it would be preferred relative to brown-seeded cultivars in human consumption markets Can. J. Anim. Sci. (2014) 94: 669678 doi:10.4141/cjas-2014-056 669

670 CANADIAN JOURNAL OF ANIMAL SCIENCE (Matus-Ca diz et al. 2003). The hairless varieties were created by mutagenicity using the parent hairy brown coloured Keet canary seed. The composition of canary seed, including the brown hairless varieties, has been reported and demonstrates a composition that is similar to cereal grains such as wheat (Abdel-Aal et al. 1997; Newkirk et al. 2011) with the exception of increased protein and ether extract. Anti-nutritional factors commonly found in grains have also been examined in canary seed and have either been found to be low and similar to other cereal grains, or non-detectable (Canary Seed Development Commission of Saskatchewan, personal communication). Levels of phytate are approximately twice as high as wheat (Abdel- Aal et al. 2011), but are well within the range of other food and feed ingredients (Classen et al. 2010). This indicates that there is low potential for the hairless canary seed to cause negative effects in feed or food applications. The digestibility of canary seed nutrients has also been studied in pigeon and poultry models (Massey et al. 1960; Bishop and Taylor 1963; Hullar et al. 1999; Newkirk et al. 2011). AME values were found to be 3508 and 3205 kcal kg 1 for pigeons and chickens, respectively, while CP and EE digestibility values were consistently high (CP 85.8% in pigeons, 88.3% in chickens; EE 94.1% in pigeons, 87.7% in chickens). As expected, based on protein digestibility, amino acids in the hairless canary seed cultivar CDC Maria are highly digestible with an average apparent ileal digestibility value of approximately 85% (Newkirk et al. 2011). Thacker (2003) found dry matter digestibility to decrease and protein digestibility to increase with increasing levels of canary seed in pig diets; the author attributed the change in dry matter digestibility to the canary seed hull. Currently, canary seed is used as a major component only in pet and wild bird feed mixtures (Nagy 1996) but other uses are under investigation (Abdel-Aal et al. 2010, 2011). Use in animal feeds appears feasible based on nutrient levels and digestibility as well as the general lack of anti-nutritional factors. Use in feed has been shown feasible in pigs (Thacker 2003) and chickens (Newkirk et al. 2011), but confirmation of feeding value and lack of negative effects is warranted. Therefore, a study was conducted to confirm the potential of hairless canary seeds as a grain ingredient in animal diets using broiler chickens as a model and also to compare the hulled yellow cultivar (C05041) with the hulled brown cultivar (CDC Maria). There are three main objectives in this study: (1) to determine the impact of graded levels of hulled yellow and brown canary seed on the performance of broiler chickens grown to 35 d, (2) to investigate the effect of different levels of hulled yellow and brown canary seed on the weight of the eviscerated carcass and various organs at 35 d, and (3) to examine the effects of graded levels of hulled yellow and brown canary seed on gross pathology and histopathology of key organs. MATERIALS AND METHODS The experimental protocol was approved by the Animal Care Committee of University of Saskatchewan. The chickens were also raised and cared for based on the Canadian Council on Animal Care (1993). Experimental Design The experimental design was a 2 4 factorial arrangement that utilized two canary seed cultivars [hulled yellow canary seed (C05041); hulled brown canary seed (CDC Maria)], and four levels of canary seed inclusion (0, 15, 30 and 45%). The experiment was completed in two stages based on experimental cage size (0 to 21 and 22 to 35 d of age). From 0 to 21 d of age, the 0% treatment was repeated twice, but because of space limitations only once from 22 to 35 d of age. Therefore, there were eight and seven dietary treatments from 0 to 21 and from 22 to 35 d of age, respectively. From 0 to 21 d of age, there were eight replications per treatment with six birds per replication (three female and three male) and from 22 to 35 d of age, there were five replications per treatment with four birds (two female and two male) per replication. Animals and Housing A total of 384 d-old male and female Ross Ross 308 broiler chickens obtained from a commercial hatchery (Lilydale, Wynyard) were randomly allocated to 64 battery brooder cages (Jamesway Manufacturing Co., Ft. Atkinson, WI; 44 cm wide, 85 cm long, 25 cm high) where they remained until 21 d of age. At 21 d of age, birds were randomly chosen from each treatment to provide two females and two males in each of five replications, and placed in grower battery cages (manufacturer unknown; 69 cm wide, 67 cm long, 35 cm high). Overall there were 140 broiler chicks used in this portion of the research. Room temperature was initially set to 358C and then gradually reduced 38C per week until 228C was reached, where it remained for the duration of the trial. The light intensity was 40 lx with 23 h of light and 1 h of darkness from 0 to 7 d of age. After 8 d of age, the light intensity was changed to 20 lx with 18 h of light and 6 h of darkness. Water and feed were provided to the birds on an ad libitum basis. Experimental Diets Starter (Table 1) and grower (Table 2) diets were fed from 0 to 14 and from 15 to 35 d of age, respectively. The canary seed was ground using a hammer mill (model160- D, Jacobson Machine Works, Minneapolis, MN) with a screen-hole size of 2.78 mm. Wheat was ground in the same hammer mill using a 4.0-mm screen-hole size. All diets were mixed using a Hobart (model L-800, Hobart Canada, North York, ON) mixer and fed in mash form. The diets were formulated to meet or exceed the Aviagen (2007) recommended values, and were formulated based on energy and amino acid digestibility values reported by Newkirk et al. (2011).

CLASSEN ET AL. * BROILER CHICKENS FED CANARY SEED 671 Table 1. Composition of broiler starter diets fed from 0 to 14 d of age Canary seed levels (%) Ingredients (%) 0 15 30 45 Wheat 56.29 41.26 25.79 10.54 Soybean meal 34.46 34.14 33.82 33.49 C05041 canary seed/ CDC 0.00 15.00 30.00 45.00 Maria canary seed Canola oil 4.24 4.77 5.30 5.83 Limestone 1.91 1.91 1.91 1.91 Di-calcium phosphate 1.48 1.49 1.50 1.50 Vitaminmineral premix z 0.50 0.50 0.50 0.50 Sodium chloride 0.36 0.36 0.36 0.36 Choline chloride 0.10 0.10 0.10 0.10 L-Lysine HCl 0.16 0.19 0.22 0.25 DL-Methionine 0.30 0.28 0.25 0.22 L-Threonine 0.11 0.14 0.17 0.20 Wheat Avizyme 1302 y 0.025 0.025 0.025 0.025 Salinomycin sodium x 0.05 0.05 0.05 0.05 Virginiamycin w 0.025 0.025 0.025 0.025 Formulated nutrient composition Crude protein 24.16 24.81 25.47 26.11 AMEn (kcal kg 1 ) 3025 3025 3025 3025 Ether extract 5.66 6.18 6.70 7.22 Lysine 1.41 1.41 1.41 1.42 Digestible lysine 1.27 1.27 1.27 1.27 Arginine 1.53 1.56 1.58 1.61 Digestible arginine 1.40 1.42 1.45 1.47 Methionine 0.63 0.60 0.58 0.56 Digestible methionine 0.60 0.57 0.55 0.53 Methionine and cysteine 1.03 1.04 1.04 1.05 Digestible methionine and 0.94 0.94 0.94 0.94 cysteine Threonine 0.95 0.97 0.98 1.00 Digestible threonine 0.83 0.83 0.83 0.83 Isoleucine 0.97 0.98 0.99 1.00 Digestible isoleucine 0.88 0.88 0.88 0.89 Leucine 1.71 1.71 1.71 1.71 Digestible leucine 1.53 1.53 1.53 1.53 Valine 1.07 1.08 1.08 1.09 Digestible valine 0.95 0.95 0.95 0.95 Calcium 1.05 1.05 1.05 1.05 Non-phytate P 0.50 0.50 0.50 0.50 z Supplied per kilogram of diet: vitamin A (retinyl acetateretinyl palmitate), 11 000 IU; vitamin D, 2200 IU; vitamin E (dl-a-topheryl acetate), 300 IU; menadione, 2.0 mg; thiamine, 1.5 mg; riboflavin, 6.0 mg; niacin, 60 mg; pyridoxine, 4 mg; vitamin B 12, 0.02 mg; pantothenic acid, 10.0 mg; folic acid, 0.6 mg; biotin, 0.15 mg; iron, 80 mg; zinc, 80 mg; manganese, 80 mg; copper, 10 mg; iodine, 0.8 mg; selenium, 0.3 mg; and CaCO 3, 500 mg. y Avizyme 1302 (Danisco Animal Nutrition, Marlborough, Wiltshire, United Kingdom). x Biocox (Alpharma Canada Corp, Mississauga, ON). w Stafac-44 (Phibro Animal Health, Ridgefield Park, NJ). Table 2. Composition of broiler grower diets fed from 15 to 35 d of age Canary seed levels (%) Ingredients (%) 0 15 30 45 Wheat 66.51 51.26 36.02 20.77 Soybean meal 26.04 25.71 25.39 25.06 C05041 canary seed/ CDC 0.00 15.00 30.00 45.00 Maria canary seed Canola oil 2.97 3.50 4.04 4.57 Limestone 1.65 1.65 1.65 1.65 Di-calcium phosphate 1.24 1.24 1.25 1.25 Vitaminmineral premix z 0.50 0.50 0.50 0.50 Sodium chloride 0.34 0.34 0.34 0.34 Choline chloride 0.10 0.10 0.10 0.10 L-Lysine HCl 0.20 0.23 0.26 0.29 DL-Methionine 0.25 0.22 0.20 0.17 L-Threonine 0.11 0.13 0.16 0.19 Wheat Avizyme 1302 y 0.025 0.025 0.025 0.025 Salinomycin sodium x 0.05 0.05 0.05 0.05 Virginiamycin w 0.025 0.025 0.025 0.025 Formulated nutrient composition Crude protein 21.67 22.31 22.96 23.61 AMEn (kcal kg 1 ) 3025 3025 3025 3025 Ether extract 4.53 5.05 5.57 6.09 Lysine 1.22 1.22 1.22 1.23 Digestible lysine 1.10 1.10 1.10 1.10 Arginine 1.31 1.34 1.36 1.39 Digestible arginine 1.19 1.21 1.24 1.26 Methionine 0.54 0.52 0.50 0.48 Digestible methionine 0.52 0.49 0.47 0.45 Methionine and cysteine 0.92 0.93 0.93 0.94 Digestible methionine and 0.84 0.84 0.84 0.84 cysteine Threonine 0.83 0.85 0.87 0.88 Digestible threonine 0.73 0.73 0.73 0.73 Isoleucine 0.84 0.85 0.86 0.87 Digestible isoleucine 0.77 0.77 0.77 0.77 Leucine 1.50 1.50 1.51 1.51 Digestible leucine 1.35 1.35 1.35 1.35 Valine 0.95 0.95 0.96 0.96 Digestible valine 0.84 0.84 0.84 0.84 Calcium 0.90 0.90 0.90 0.90 Non-phytate P 0.45 0.45 0.45 0.45 z Supplied per kilogram of diet: vitamin A (retinyl acetateretinyl palmitate), 11 000 IU; vitamin D, 2200 IU; vitamin E (dl-a-topheryl acetate), 300 IU; menadione, 2.0 mg; thiamine, 1.5 mg; riboflavin, 6.0 mg; niacin, 60 mg; pyridoxine, 4 mg; vitamin B 12, 0.02 mg; pantothenic acid, 10.0 mg; folic acid, 0.6 mg; biotin, 0.15 mg; iron, 80 mg; zinc, 80 mg; manganese, 80 mg; copper, 10 mg; iodine, 0.8 mg; selenium, 0.3 mg; and CaCO 3, 500 mg. y Avizyme 1302 (Danisco Animal Nutrition, Marlborough, Wiltshire, United Kingdom). x Biocox (Alpharma Canada Corp, Mississauga, ON). w Stafac-44 (Phibro Animal Health, Ridgefield Park, NJ). Data Collection Feed intake and body weight were measured on a pen basis at 0, 7, 14, 21, 28 and 35 d of age. Mortality was recorded daily and the dead birds were sent to an independent laboratory (Prairie Diagnostic Service, 52 Campus Drive, Saskatoon, SK) for necropsy. At 35 d of age, the birds were individually weighed, and then stunned (VS 200 stunner knife, Midwest Processing System, Edina, MN) and bled by hand-cutting the carotid artery and jugular veins on both sides of the neck. After a 2-min exsanguination period, the birds were scalded in a water tank at 66 to 688C for 30 s and then defeathered in an automatic feather picker (Pro model K7080, Featherman Equipment, Jamesport, MO) for another 30 s. The birds were then eviscerated to obtain carcass weight. Additionally, the gizzard, small intestine

672 CANADIAN JOURNAL OF ANIMAL SCIENCE (jejunum and ileum), heart, liver, kidney, bursa of Fabricius and spleen were collected and weighed. Intestinal contents were removed prior to being weighed. A complete necropsy of 140 broiler chickens was conducted at 35 d of age. Body condition and the skeletal system and skin were examined prior to opening birds for internal examination. The air sacs, liver, heart, bursa of Fabricius, thymus, spleen, intestines, hock joint and growth plate of the tibotarsus were examined for inflammatory, neoplastic or degenerative changes. Tissue sections from the liver, heart, spleen, thymus and bursa of Fabricius (four birds per treatment) were fixed in 10% neutral buffered formalin, embedded in paraffin, sectioned at 5 mm thickness and stained with hematoxylin and eosin for histopathological examination. Statistical Analyses Statistical analyses for growth performance and cause of mortality (0 to 21 and 22 to 35 d) and organ weights (35 d) were completed using SAS software version 9.2 (SAS Institute Inc. 2001) using Proc mixed as a 42 factorial arrangement. Pseudo-replication of the 0% inclusion levels was used for the organ weights and 22- to 35-d broiler performance. Means were separated using least square means (LSMEANS) and pdmix800 (Saxton 1998). Proc Reg and Proc RSReg of SAS software version 9.2 (SAS Institute Inc. 2001) (linear and quadratic regression analyses, respectively) were also used to establish relationships between response criteria and graded levels of canary seeds. The linear (1) and quadratic (2) functions were: Y b 0 b 1 C (1) Y b 0 b 1 C b 2 C 2 (2) Where Y is predicted response, C is the canary seed level of the diet, b 0 is intercept, and b 1 and b 2 are regression coefficients. Unless otherwise stated, the level of significance was set at P50.05. s 0.05 but 50.10 are reported in tables. RESULTS Broiler Performance The main effect of level of canary seed affected aspects of broiler performance, while cultivar only impacted the incidence of mortality. There were no significant interactions between cultivar and dietary level of canary seed inclusion for broiler performance criteria and therefore only the main effects are reported. During the 0- to 21-d portion of the trial, level of canary seed inclusion affected body weight (14 and 21 d of age) and body weight gain (814, 1521 and 021 d of age) in a quadratic manner (Table 3). Growth for the 0% inclusion rate was lowest but not different from the 45% treatment; growth was maximum for either the 15 or 30% treatments depending on the time period. Similarly, a quadratic response was seen for body weight at 28 and 35 d of age, but in this case broilers fed the 0% treatment weighed less than birds from all treatments with added canary seed (Table 4). Body weight gain from 22 to 28 d of age resulted in a positive linear response with increasing diet canary seed inclusion. Table 3. The effects of dietary canary seed level and cultivar on body weight and body weight gain in broiler chickens from 0 to 21 d of age Body weight (kg) Body weight gain (kg) Nutrient content 0 d 7 d 14 d 21 d 07 d 814 d 1521 d 021 d 0% 0.047 0.150 0.388 0.817c 0.104 0.238 0.429c 0.771c 15% 0.047 0.157 0.418 0.885ab 0.110 0.261 0.467a 0.838ab 30% 0.047 0.163 0.428 0.892a 0.116 0.265 0.464ab 0.845a 45% 0.047 0.156 0.394 0.830bc 0.109 0.238 0.436bc 0.783bc Intercept 0.387 0.817 0.237 0.430 0.770 C 0.0033 0.0068 0.0025 0.0034 0.0068 C 2 0.00007 0.00014 0.000055 0.000074 0.00014 R 2 0.088 0.135 0.068 0.148 0.135 L NS z NS NS 0.029 NS NS 0.018 0.028 Linear NS NS NS NS NS NS NS NS Quadratic NS NS 0.020 0.003 NS 0.039 0.002 0.003 C05041 0.047 0.155 0.405 0.860 0.108 0.251 0.454 0.813 CDC Maria 0.047 0.158 0.408 0.852 0.111 0.250 0.444 0.805 C NS NS NS NS NS NS NS NS Pooled SEM 0.00020 0.0022 0.0068 0.011 0.0022 0.0060 0.0055 0.011 LC NS NS NS NS NS NS NS NS z NSP0.10. ac Means within a column and main effect with different letters are significantly different (P50.05).

CLASSEN ET AL. * BROILER CHICKENS FED CANARY SEED 673 Table 4. The effects of dietary canary seed level and cultivar on body weight and body weight gain in broiler chickens from 22 to 35 d of age Body weight (kg) Body weight gain (kg) 28 d 35 d 2228 d 2935 d 2235 d 0% 1.425b 2.097b 0.606b 0.674 1.278 15% 1.573a 2.184a 0.689a 0.610 1.299 30% 1.561a 2.218a 0.671a 0.656 1.328 45% 1.532a 2.175a 0.700a 0.644 1.343 Intercept 1.428 2.093 0.625 C 0.011 0.0086 0.0018 C 2 0.00021 0.00015 R 2 0.410 0.246 0.149 L B0.0001 0.022 0.017 NS z NS Linear 0.014 0.036 0.014 NS 0.066 Quadratic 0.0002 0.014 NS NS NS C05041 1.511 2.153 0.649 0.643 1.291 CDC Maria 1.535 2.184 0.684 0.650 1.333 C NS NS NS NS NS Pooled SEM 0.0146 0.0147 0.013 0.010 0.014 LC NS NS NS NS NS z NSP0.10. ac Means within a column and main effect with different letters are significantly different (P50.05). Feed intake responded to dietary level of canary seed in a quadratic manner from 15 to 21 and from 0 to 21 d of age (Table 5). In both cases, the lowest value was found for the 45% treatment but it was not significantly different than for birds in the 0% treatment; feed intake values for birds from the 15 and 30% treatments were highest. A similar quadratic response was seen for feed intake from 22 to 28 d of age (Table 6). There was a linear decrease in mortality corrected feed to gain ratio with increasing level of canary seed in the diet from 0 to 7, 15 to 21, 0 to 21, 22 to 28 and 22 to 35 d of age (Tables 5 and 6). Mortality only occurred during the 0- to 21-d period and the results are shown in Table 7. Level of canary seed inclusion in the diet did not affect death loss, but mortality was higher for birds fed the C05041 canary seed in contrast to those fed CDC Maria canary seed. Significant differences were seen between the two cultivars in the overall infection mortality from 0 to 7 d of age (Table 8) and from 0 to 21 d of age (Table 9). Level of canary seed did not affect cause of mortality. Organ Weights Broiler organ weights at 35 d of age are presented on an absolute weight (g) and percent of live weight basis (Tables 10 and 11). These criteria were not affected by genotype or the interaction between genotype and level of canary seed inclusion. Body weight was affected by Table 5. The effects of dietary canary seed level and cultivar on feed intake and mortality corrected feed to gain ratio in broiler chickens from 0 to 21 d of age Feed intake (kg) Feed to gain 07 d 814 d 1521 d 021 d 07 d 814 d 1521 d 021 d 0% 0.113 0.334 0.623ab 1.071ab 1.093a 1.386 1.456a 1.383a 15% 0.115 0.349 0.657a 1.128a 1.054b 1.308 1.413b 1.335b 30% 0.120 0.348 0.647a 1.130a 1.036bc 1.325 1.378bc 1.306c 45% 0.109 0.314 0.591b 1.020b 1.007c 1.355 1.349c 1.290c Intercept 0.623 1.068 1.088 1.453 1.375 C 0.0038 0.0073 0.0018 0.0024 0.0021 C 2 0.00010 0.00019 R 2 0.168 0.130 0.254 0.371 0.512 L NS z NS 0.009 0.029 0.0004 NS B0.0001 B0.0001 Linear NS NS NS NS B0.0001 NS B0.0001 B0.0001 Quadratic NS 0.075 0.003 0.0068 NS 0.059 NS 0.057 C05041 0.113 0.337 0.640 1.101 1.056 1.344 1.407 1.335 CDC Maria 0.115 0.336 0.619 1.074 1.039 1.343 1.392 1.322 C NS NS NS NS NS NS NS NS Pooled SEM 0.0022 0.0068 0.0079 0.016 0.0077 0.014 0.0083 0.0061 LC NS NS NS NS NS NS NS NS z NSP0.10. ac Means within a column and main effect with different letters are significantly different (P50.05).

674 CANADIAN JOURNAL OF ANIMAL SCIENCE Table 6. The effects of dietary canary seed level and cultivar on feed intake and mortality corrected feed to gain ratio in broiler chickens from 22 to 35 d of age Feed intake (kg) Feed to gain 2228 d 2935 d 2235 d 2228 d 2935 d 2235 d 0% 0.957 1.095 2.052 1.598a 1.628 1.610 15% 1.000 1.099 2.098 1.461b 1.826 1.618 30% 0.986 1.084 2.070 1.478b 1.688 1.561 45% 0.956 1.069 2.024 1.377b 1.663 1.515 Intercept 1.579 1.627 C 0.0044 0.0023 C 2 R 2 0.255 0.195 L 0.083 NS NS 0.0026 NS 0.017 Linear NS z NS NS 0.0009 NS 0.0043 Quadratic 0.010 NS 0.067 NS 0.097 NS C05041 0.973 1.081 2.054 1.511 1.685 1.593 CDCMaria 0.976 1.092 2.068 1.446 1.717 1.559 C NS NS NS 0.091 NS NS Pooled SEM 0.0075 0.0070 0.013 0.023 0.033 0.014 LC NS NS NS NS NS NS z NSP0.10. a, b Means within a column and main effect with different superscripts are significantly different (P50.05). dietary level of canary seed and therefore many absolute weights were similarly affected. Carcass, gizzard, ileum, heart and spleen weights responded in a quadratic fashion with the 0% level having the lowest weights and the 15 and 30% inclusion levels having the highest values. In general, birds fed diets with canary seed had higher carcass and organ weights than those fed the 0% control diet. Liver weight increased in a linear fashion with increasing dietary level of canary seed. When weights are expressed on a percentage of live weight basis, no effects of level of canary seed were found for carcass (P 0.06), gizzard, ileum, jejunum and ileum, heart, liver and spleen. Data for jejunum, kidney and bursa of Fabricius weights demonstrate a quadratic response with the highest proportional weights for the control and 45% inclusion levels. Gross Pathology and Histopathology All birds that were on trial were examined for gross pathological lesions at trial end and tissues from four randomly selected birds per treatment were examined for histopathological lesions. Gross necropsy demonstrated that six of 140 birds had arthritis or tendinitis in the hock joint and three of 140 birds had tibial dyschondroplasia, but no relationship to treatment was found. No gross or histopathological lesions were seen Table 7. The effects of dietary canary seed level and cultivar on mortality in broiler chickens from 0 to 21 d of age Mortality (%) 07 d 814 d 1521 d 021 d 0% 2.08 1.04 0.00 3.13 15% 4.17 1.04 1.04 6.25 30% 4.17 1.04 1.04 7.29 45% 1.04 1.04 2.08 3.13 Intercept C C 2 R 2 L NS z NS NS NS Linear NS NS NS NS Quadratic NS NS NS NS C05041 4.69a 1.04 1.56 7.29a CDC Maria 1.04b 1.04 0.52 2.60b C 0.049 NS NS 0.041 Pooled SEM 0.950 0.508 0.508 1.213 LC NS NS NS NS z NSP0.10. a, b Means within a column and main effect with different letters are significantly different (P50.05). in the liver, heart, spleen, bursa of Fabricius and thymus in any of the birds examined. DISCUSSION The results of this research confirm that high levels of hairless hulled canary seed can be fed to broiler chickens without adverse effects on growth, feed intake, feed efficiency or mortality. In the research of Newkirk et al. (2011), 50% canary seed levels in broiler diets did not affect production traits and in the present trial 45% dietary inclusion resulted in performance that equalled or exceeded the wheat control diet (0% canary seed). Quadratic relationships between dietary level of canary seed and production criteria in the present research suggest that intermediate levels of canary seed enhance broiler performance, but a reason for such an effect is not obvious. It is possible that a slightly better nutrient balance in these diets could have resulted in this effect. The cultivar of hairless hulled canary seed did not affect any production traits except early mortality; more birds fed the C05041 cultivar died than broilers fed the CDC Maria cultivar. Death loss differences were due to infectious causes and these differences were significant for the 0- to 7-d and 0- to 21-d time periods. Closer examination of the data reveals that much of the death loss was due to yolk sac infection, which occurs within the first week of life and is attributed to breeding flock and hatchery sanitation, and not to farm conditions.

CLASSEN ET AL. * BROILER CHICKENS FED CANARY SEED 675 Table 8. The effects of dietary canary seed level and cultivar on cause of mortality in broiler chickens from 0 to 7 d of age Infection YSI z Osteo. z Endo. Total SDS z Unknown Starvation Dehydration 0% 2.08 0.00 0.00 2.08 0.00 0.00 0.00 0.00 15% 2.08 1.04 0.00 3.13 0.00 0.00 0.00 1.04 30% 1.04 0.00 0.00 1.04 0.00 1.04 2.08 0.00 45% 0.00 0.00 0.00 0.00 0.00 0.00 1.04 0.00 Intercept C C 2 R 2 L NS y NS NS NS NS NS NS NS Linear NS NS NS NS NS NS NS NS Quadratic NS NS NS NS NS NS NS NS C05041 2.60 0.52 0.00 3.13a 0.00 0.52 0.52 0.52 CDC Maria 0.00 0.00 0.00 0.00b 0.00 0.00 1.04 0.00 C 0.057 NS NS 0.026 NS NS NS NS Pooled SEM 0.675 0.260 0.000 0.716 0.000 0.260 0.444 0.260 LC NS NS NS NS NS NS 0.084 NS z YSI, yolk sac infection; Osteo., osteomyelitis; Endo., endocarditis; SDS, sudden death syndrome. y NSP0.10. a, b Means within a column and main effect with different letters are significantly different (P50.05). Table 9. The effects of dietary canary seed level and cultivar on cause of mortality in broiler chickens from 0 to 21 d of age Infection YSI z Osteo. z Endo. z Total SDS z Unknown Starvation Dehydration 0% 2.08 0.00 0.00 2.08 1.04 0.00 0.00 0.00 15% 2.08 1.04 0.00 3.13 2.08 0.00 0.00 1.04 30% 1.04 0.00 1.04 2.08 2.08 1.04 2.08 0.00 45% 0.00 1.04 0.00 1.04 1.04 0.00 1.04 0.00 Intercept C C 2 R 2 L NS y NS NS NS NS NS NS NS Linear NS NS NS NS NS NS NS NS Quadratic NS NS NS NS NS NS NS NS C05041 2.60 1.04 0.52 4.17a 1.56 0.52 0.52 0.52 CDC Maria 0.00 0.00 0.00 0.00b 1.56 0.00 1.04 0.00 C NS NS NS 0.02 NS NS NS NS Pooled SEM 0.675 0.365 0.260 0.87 0.612 0.260 0.444 0.260 LC NS NS NS NS NS NS 0.084 NS z YSI, yolk sac infection; Osteo., osteomyelitis; Endo., endocarditis; SDS, sudden death syndrome. y NSP0.10. a, b Means within a column and main effect with different letters are significantly different (P50.05).

676 CANADIAN JOURNAL OF ANIMAL SCIENCE Table 10. The effects of dietary canary seed level and cultivar on broiler live, carcass and organ weights (g) at 35 d of age LW z Carcass Gizzard Jejunum Ileum JI z Heart Liver Kidney BF z Spleen 0% 2068b 1533b 24.4b 22.5 18.3 40.8 11.9 42.0 14.6 4.4 1.9b 15% 2213a 1649a 26.4a 22.5 18.7 41.3 12.9 42.9 14.6 4.1 2.2ab 30% 2188a 1626a 26.6a 22.8 19.6 42.4 13.2 45.0 14.6 4.1 2.5a 45% 2172a 1615a 26.1a 22.8 18.3 41.0 12.7 44.9 14.8 4.2 2.2ab Intercept 2073 1535 24.6 18.1 11.9 42.1 1.9 C 10.475 8.590 0.160 0.0969 0.094 0.072 0.038 C 2 0.190 0.157 0.0028 0.002 0.0017 0.0007 R 2 0.054 0.059 0.050 0.030 0.050 0.033 0.053 L 0.026 0.021 0.040 NS y 0.089 NS 0.070 NS NS NS 0.032 Linear 0.078 0.076 0.053 NS NS NS 0.089 0.022 NS NS 0.052 Quadratic 0.018 0.012 0.037 NS 0.034 NS 0.024 NS NS NS 0.030 C05041 2134 1583 25.3 22.4 18.7 41.0 12.6 43.1 14.5 4.0 2.1 CDC Maria 2187 1628 26.4 22.9 18.8 41.7 12.7 44.3 14.8 4.4 2.3 C NS NS 0.066 NS NS NS NS NS NS 0.095 NS Pooled SEM 18.27 14.22 0.303 0.259 0.214 0.439 0.172 0.527 0.176 0.096 0.070 LC NS NS NS NS NS NS NS NS NS NS NS z LW, live weight; JI, jejunum and ileum; BF, bursa of Fabricius. y NSP0.10. a, b Means within a column and main effect with different letters are significantly different (P50.05). Early yolk sac infection has also been shown to lead to other infectious disease loss later in the flock and this was seen for the C05041 birds, which had numerically higher levels of osteomyelitis and endocarditis. Because of the timing and nature of the mortality, it is concluded that this difference in mortality is more likely a chance Table 11. The effects of dietary canary seed level and cultivar on broiler live, carcass and organ weights (% of live weight) at 35 d of age Carcass Gizzard Jejunum Ileum JI z Heart Liver Kidney BF z Spleen 0% 74.0 1.18 1.09a 0.89 1.98 0.58 2.04 0.70 0.21a 0.09 15% 74.6 1.20 1.01b 0.85 1.86 0.58 1.94 0.66 0.18b 0.10 30% 74.3 1.23 1.04ab 0.90 1.94 0.60 2.05 0.67 0.19b 0.11 45% 74.3 1.20 1.05ab 0.85 1.90 0.58 2.08 0.68 0.19ab 0.10 Intercept 1.088 0.701 0.213 C 0.0052 0.0030 0.0023 C 2 0.00010 0.000058 0.000042 R 2 0.042 0.040 0.050 L NS y NS 0.041 0.065 0.066 NS 0.068 0.063 0.033 NS Linear NS NS NS NS NS NS NS NS 0.098 NS Quadratic 0.064 NS 0.021 NS NS NS NS 0.027 0.021 NS C05041 74.2 1.19 1.05 0.88 1.93 0.59 2.03 0.68 0.19 0.10 CDC Maria 74.4 1.21 1.05 0.86 1.91 0.58 2.03 0.68 0.20 0.11 C NS NS NS NS NS NS NS NS NS NS Pooled SEM 0.098 0.013 0.0098 0.0083 0.0160 0.0057 0.0190 0.0059 0.0041 0.0032 LC NS NS NS NS NS NS NS NS NS NS z JI jejunum and ileum; BF bursa of Fabricius. y NSP0.10. a, b Means within a column and main effect with different letters are significantly different (P50.05).

CLASSEN ET AL. * BROILER CHICKENS FED CANARY SEED 677 effect rather than a biological effect associated with one cultivar. This is supported by the total lack of mortality from 21 to 35 d of age for the entire flock and the lack of a cultivar or level of canary seed effect on gross pathology and histopathology. Overall, the data from this trial indicate that differences in nutritional quality between the brown-seeded CDC Maria and the yellow-seeded C05041 are minimal in regard to safety and efficacy in animal feeding. Differences were found in carcass and organ weights due to diet level of canary seed, but these reflected the heavier body weights at 35 d for the birds fed canary seed. As such, it does not add to the understanding of canary seed feeding value. Values in proportion to live weight permit a clearer understanding and for the most part are not affected by either cultivar or dietary canary seed level. Exceptions are a quadratic response in jejunum weight with values tending to be lower for birds fed canary seed. Newkirk et al. (2011) found a numerical reduction in both jejunum and ileum weight for birds fed CDC Maria vs wheat-based diets and attributed this to the more digestible nature and lower soluble fibre content of canary seed in comparison to wheat. High digesta viscosity associated with more soluble fibre has been shown to decrease intestinal mobility and increase microbial activity, which in turn increases intestinal growth and weight (Salih et al. 1991; Brenes et al. 1993; Choct et al. 1996). In the present work, there was no effect of diet level on ileum and the combined jejunum and ileum weights. Kidney and bursa of Fabricius weights were also affected in a quadratic manner by level of canary seed, but the 0 and 45% values are not significantly different. Lower proportional weights for the intermediate treatments are in contrast to the heavier body weights for these treatments. It may be that birds in these treatments are slightly more advanced from a physiological development perspective and because bursa of Fabricius weights decline with age, it is possible that this accounts for the relatively small but significant differences. Newkirk et al. (2011) did not find any differences in organ weights for birds fed CDC Maria or wheat-based diets. These results in combination with the lack of histopathological lesions in this research support the conclusion that feeding canary seed does not negatively affect animal well-being. No inflammatory or degenerative lesions were seen in the liver, heart, bursa of Fabricius, spleen and thymus in any of the treatment groups. Lesions in the joint and tendons (locally extensive arthritis or tendinitis) are likely individual problems due a mechanical injury. Similarly, it is highly probable that tibial dyschondroplasia is an incidental finding. The failure to associate these lesions with specific treatments supports this conclusion. Previous research (Newkirk et al. 2011) found an increase is gizzard lesions associated with feeding hulled canary seed cultivars, but gizzard lesions were not noted in the present research. The results of the present research, from performance, carcass and organ, and pathology perspectives, indicate that hairless hulled canary seed has merit and is safe for poultry feeding. Further, this research demonstrates that the yellow seed coated cultivar (C05041) is equal to the brown seed coated cultivar in feeding value. ACKNOWLEDGEMENTS The authors acknowledge research funding by the Canaryseed Development Commission of Saskatchewan and by the Agriculture Council of Saskatchewan through the Advancing Canadian Agriculture and Agri-Food Saskatchewan (ACAAFS) program. 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