Effects of 50 Percent Substitution of Soybean Meal by Alternative Proteins from <i>Hermetia illucens</i>or Spirulina platensis in Meat-Type Chicken Diets with Graded Amino Acid Supply

Alternative protein sources, such as insects or algae meals are in special focus of animal nutrition in order to replace soybean meal (SBM). As part of the multidisciplinary project “sustainability transitions” this study evaluated effects of replacing SBM by partly defatted larvae meal from the black soldier fly, Hermetia illucens or meal from the micro algae Spirulina platensis in broiler diets. The aim of the current study was to investigate the chickens’ growth performance and the intestinal morphology as well as the health status. 288 one-day-old male growing chickens (Ross 308) from a commercial hatchery were randomly allotted to 48 pens (6 birds per pen) for the growth study with five diets and feed supply on free choice level. The control diet was based on wheat, corn and SBM. The experimental diets replaced 50% of SBM by the alternative proteins under study, both on a basic level of amino acid (AA) supplementation (Lys and Met added equal to the control diet) (diet HM and SM) and an extended level of AA fortification (Lys, Met, Thr, Arg, Val added) (diet HM+ and SM+). Response of chickens was evaluated by zoo-technical parameters, which were under weekly control (e.g. growth, feed intake, feed conversion ratio, mortality). After finishing the growth study birds from control and the experimental diets at the basic level of AA supplementation were slaughtered after 12 hour fastening and utilized for gut morphometric analysis and histological evaluation of the health status. Diets at a basic level of AA fortification led to significant depression of growth, feed intake, feed conversion ratio, and protein conversion ratio, respectively. In addition, the acceptance of the Spirulina diet was lower (p ≤ 0.001) as compared to the Hermetia based diet. However, the extended level of AA supplementation improved all parameters significantly. Diets without extended AA supply altered some morphological parameters of the intestinal wall, but the nutritional significance of this observation needs to be verified in AA balanced diets. The health state of chicken was not impaired by the diets under study.

Assessing Lysine Requirement of Growing Chicken by Direct Comparison between Supplementation Technique and “Goettingen Approach”

Validated procedures play an important role to obtain accurate information about individual amino acid requirement data. The aim of the present study was to assess lysine (Lys) requirement of growing chicken both by classical supplementation technique and principles of diet dilution technique as applied with “Goettingen approach”. During the starter period (1 - 21 d), a growth study with male meat type chicken (Ross 308) was conducted making use of five graded dietary Lys-levels (3 repetition boxes with 3 birds/box). L-Lys×HCl was gradually added to a diet based on wheat, soybean protein concentrate, wheat gluten and fishmeal to yield 80%, 87.5%, 95%, 102.5% and 110% of the expected requirement level (13 g Lys/kg as fed). Diets were iso-energetic (12.8 MJME/kg) and iso-nitrogenous (21.65% crude protein). Birds were fed on free choice level also to assess the feed intake (FI) effects as important factor on traditional response criteria. Analyzed body composition at start and end of the growth study yielded N deposition (ND) data for further data assessment using exponential approximations depending on dietary Lys content or observed Lys intake. The results indicated significant differences (p < 0.05) in response on body weight gain (BWG) and observed dietary protein quality with unexpected consequences for the derived Lys requirement data. According to the independent variable (Lys in % of diet versus daily Lys intake) and aimed level of daily ND, the needed in-feed content of Lys varied between 1.24% and 1.46%. Application of the exponential modelling by “Goettingen approach” overcame these misleading conclusions by modelling the relationship between required Lys intake and observed response data (BWG, ND) taking also into account the expected real feed intake to formulate the needed in-feed concentration.

Age and Gender Dependent Amino Acid Concentrations in the Feather, Feather-Free and Whole Empty Body Protein of Fast Growing Meat-Type Chickens

Two consecutive growth experiments with meat-type chickens (Ross 308) were conducted in order to quantify the age-dependent amino acid (AA) content in the whole body protein of male and female birds based on experimental data of the feather and feather-free body protein fractions. Birds were reared under uniform housing and feeding conditions (floor pens, 15 pens per gender, 5 birds per pen) during the starter (day 1 to 22) and grower period (day 22 to 36). Both the starter and grower diet based on corn, wheat, soybean meal, soybean protein concentrate and feed amino acids was formulated to ensure an equal feed protein quality close to the ideal amino acid ratio by adjusting a constant mixture of the feed proteins. At start of the experiment and further on weekly up to the end of the 5th week, 15 birds per gender (each 3 pens of 5 birds) were selected and fasted for 24 h, to emptying of gastro-intestinal tract, respectively. Subsequently, birds were euthanized and the feathers were manually removed. Nitrogen (N) and AA content were determined both in the feather and feather-free body fraction. The concentration of individual AAs in both of body protein fraction is varying considerably. Explicitly higher Cys, Ser and Pro but importantly lower Met, Lys and His concentrations were found in the feather protein. Furthermore, significant differences (p for nearly all AAs of the studied body protein fractions and the whole empty body protein dependent on age of birds were observed. Especially high deviations were obtained during the first week of age and at the end of the experiment. According to this observed variation of AA concentrations must be concluded that the body AA composition of meat-type chickens during growth is not constant. The detected gender-specific differences for several AAs in the feather and body protein of male and female birds were rather low and with very low variation.

Age and Gender Dependent Nutrient Composition of Feather and Feather-Free Body Fractions in Meat-Type Chickens

The objective of this research was to update current results about the nutrient composition of broiler chickens during the growth period up to market age. Two growth experiments were conducted for assessing the nutrient content of feathers and feather-free body of meat-type chickens (Ross 308). Both male and female birds were reared under uniform management conditions (floor pens;15 pens per gender;5 birds per pen). Experimental diets both for the starter (day 1 to 22) and the grower period (day 22 to 36) were based on corn, wheat, soybean meal, soybean protein concentrate and well balanced with feed amino acids. The feed protein quality was adapted to the ideal amino acid ratio and equated within both of the feeding periods by adjusting a constant mixture of the feed proteins. Each 15 birds per gender (3 pens of 5 birds) were selected and subsequently fasted for 24 h before quantitative de-feathering both at start of the experiment and further on weekly up to the end of the 5th week. Nutrient content was determined in representative samples of the feather and feather-free body fraction. In the feather dry matter (DM) very high crude protein (CP) concentrations (>96%) with low age-dependent and insignificant gender-specific differences were observed. In spite, a relatively high variation of CP content in the DM of feather-free body was found. Depending on age, the body CP significantly decreased with increasing age, but male birds yielded higher (p < 0.001) CP content. The crude lipid content of the feather-free and whole empty body significantly increased with age and was higher in female as compared to male birds (p < 0.001). Depending on age and gender, the crude ash content both in feathers and feather-free body of modern fast-growing chickens was rather low and with very low variation.

Validation of the Lysine Requirement as Reference Amino Acid for Ideal In-Feed Amino Acid Ratios in Modern Fast Growing Meat-Type Chickens

Nitrogen (N) balance studies were conducted with male growing broiler chickens to reevaluate the lysine (Lys) requirement of a modern broiler strain (Ross 308), making use of eight diets with graded crude protein (CP) supply (6% - 34% CP as-fed). Wheat, soy protein concentrate, wheat gluten, fishmeal and crystalline amino acids (AAs) were the protein sources in the experimental diets with Lys as limiting AA. Following an adaptation period of five days, two consecutive excreta collection periods (2 × 5 d) were conducted: 10 - 20 d of age (starter period) and 25 - 35 d of age (grower period). Statistical evaluation of N balance data utilized an exponential modelling approach. Based on different dietary Lys efficiency, Lys requirement data were derived by modelling depending on average body weight (BW) during starter and grower period and targeted body protein deposition (PD), respectively. In addition, the influence of graded feed intake was taken into account. For the starter period at 600 g BW and assumed 10 g daily body PD, Lys requirement data between 741 mg and 823 mg per day were observed. The corresponding Lys in-feed concentration was 1.06% and 1.18%, dependent on supposed Lys efficiency at 70 g daily feed intake. For the grower period (average BW 1800 g), 1272 mg to 1473 mg Lys per day was needed to yield 16.5 g daily PD. The corresponding required Lys in-feed concentration was between 0.85% and 0.94% Lys for 150 g daily feed intake.

An Elevated Dietary Cysteine to Methionine Ratio Does Not Impact on Dietary Methionine Efficiency and the Derived Optimal Methionine to Lysine Ratio in Diets for Meat Type Chicken

Optimal dietary methionine (Met) to lysine (Lys) ratio in presence of elevated dietary cysteine (Cys) levels was derived for meat type growing chicken. Twelve averaged weighed Ross 308 birds (each 50% of male and female per dietary treatment) were utilized in N balance trials. During starter (d10 - 20) and grower period (d25 - 35) five dietary treatments were used. Diets based on uniform mixtures of maize, wheat, soybean meal, potato protein and fish meal were supplemented with crystalline amino acids (AA). In diets 1 - 3, the dietary Cys to Met ratio was set as 85, 95 and 105 to 100, respectively. Diet 4, at a Cys to Met ratio of 105 to 100, was additionally supplemented with betaine (BET) as methyl group donor. Diets 1 - 4 were limiting in Met, diet 5 without L-Lys·HCl addition was limiting in Lys. Individual N-balance data per treatment were utilized for assessing protein quality and efficiency of dietary Met (Diets 1 - 4) or Lys (Diet 5) based on “Goettingen approach”. Elevated dietary Cys supply and supplemented BET failed to improve both dietary protein quality and Met efficiency. The established optimal Met to Lys ratio was on average 34 to 100 for growing chicken during starter and grower period, respectively.

Improving the Dietary Protein Quality by Amino Acid Fortification with a High Inclusion Level of Micro Algae (<i>Spirulina platensis</i>) or Insect Meal (<i>Hermetia illucens</i>) in Meat Type Chicken Diets

The objective of this study was to measure protein quality parameters (PPV, NPU and NPUstd) of chicken diets with complete substitution of soybean meal (SBM) by Spirulina meal (SM) or partly defatted Hermetia meal (HM). N balance experiments were based on the quantitative excreta collection method, divided into starter period (10 - 20 d) and grower period (25 - 35 d). The study utilized 70 all male meat type chicken (Ross 308). Data assessment applied the exponential N utilization model of the “Goettingen approach”. The control diet was based on wheat, corn and SBM. In four experimental diets SBM was completely substituted by SM or HM, but fortified with feed amino acids (AA) both on a basic level of supplementation (Lys and Met added equal to the control diet) and on an extended level (Lys, Met, Thr, Arg, Val, Ile, His added). At a basic level of AA supplementation, complete replacement of SBM by SM or HM in chicken diets depressed dietary protein quality significantly (p 0.05). However, the extended level of AA supplementation improved protein quality parameters of the diets with both of the alternative proteins significantly (p < 0.05), but still generally not on par with the control diet. The observed responses were accentuated when the well-known effect of N intake on protein utilization was eliminated through the standardization of N intake by application of the “Goettingen approach”.

Effects of the Partial Replacement of Soybean Meal by Insect or Algae Meal in Chicken Diets with Graded Amino Acid Supply on Parameters of Gut Microbiology and Dietary Protein Quality

Insects or algae are considered future solutions to substitute imported feed proteins like soybean meal (SBM) in animal nutrition. The objective of this research, as part of the multidisciplinary project “Sustainability transitions”, focused on replacing 50% SBM by partly defatted larvae meal from black soldier flies (Hermetia illucens) or the blue-green algae Spirulina platensis in meat type chicken diets. The current study aimed to evaluate the effects on body composition, protein quality, apparent precaecal digestibility (apcD) and parameters of intestinal microbiota. In total, 288 one-day-old male growing chickens (Ross 308) from a commercial hatchery were randomly allotted to 48 pens (6 birds per pen) across five diets, which were fed at a free choice level. The control diet utilized SBM, wheat and corn as main ingredients, and experimental diets replaced 50% of SBM by the alternative proteins under study. Amino acid (AA) supplementation of the final diets was conducted both at a basic level (diets HM, SM;Lys and Met added equal to the control diet) and an extended level of AA fortification (diets HM+, SM+;extended supplementation of Lys, Met, Thr, Arg). After finishing the growth study, 4 individual birds per diet were slaughtered for whole body analyses to derive nutrient utilization and dietary protein quality parameters. Additionally, pooled chyme samples from 16 birds per diet (control, HM and SM) were analyzed to assess apcD and microbial parameters. Diets HM and SM with a basic level of AA fortification led to significant depressions in nutrient deposition and dietary protein quality. However, HM+ and SM+ diets with an extended level of AA supplementation led to significantly improved responses;however these were still generally not on par with the control diet. The HM+ diet provided superior CP deposition and dietary protein quality, as compared to the SM+ diet, but equal to the control diet. Results of gut microbiology yielded no significant effects due to feeding the alternative protein sources under study.

日龄、性别、蛋白质沉积和日粮赖氨酸效率对快速生长肉鸡的赖氨酸需求的影响

通过氮平衡试验,可以根据粗蛋白质(Crude Protein,CP)沉积量和日粮赖氨酸利用率,评估不同性别和日龄的快速生长型肉鸡(科宝500)的赖氨酸需要量。试验分为3个日龄段(第1日龄段10～25日龄,第2日龄段30～45日龄;第3日龄段50～65日龄),每个日龄段各72羽试验鸡(公母各36羽)。试验日粮由高蛋白质(HighProtein,HP)-豆粕、小麦蛋白质和结晶氨基酸〔L-苏氨酸(Thr)、DL-蛋氨酸(Met)、L-精氨酸(Arg)〕组成,共设6个级别的粗蛋白质水平,从而使得赖氨酸(4.30gLys/100gCP)成为日粮的第一限制性氨基酸〔Lys∶(Met+Cys)∶Thr∶Arg=1∶1.01∶0.91∶1.14〕。由于应用了非线性氮利用率模型,将氮维持需要量(Nitrogen Maintenance Requirement,NMR)和每日氮日沉积的理论最大值(Theoretical Maximum for day Nitrogen Retention,NRmaxT)确定为模型参数,以便能进一步评估依赖于日龄、性别和每日粗蛋白质沉积量的赖氨酸需要量。例如,按每日粗蛋白质沉积量理论潜能的60%计算得到的日粮赖氨酸需求水平(10～25日龄:赖氨酸含量1.10%,日采食量60g;30～45日龄:赖氨酸含量1.03%,日采食量140g;50～65日龄:赖氨酸含量0.96%,日采食量170g)与已报道的结果完全一致。然而,当氨基酸需要量水平确定后,采食量预测值就成为最重要的影响因子之一。而作为影响氨基酸需要量重要因素的每日粗蛋白质沉积量和日粮氨基酸的利用效率,在今后的氨基酸需要量研究中还需作进一步的研究。

Age and Gender Dependent Deposition of Crude Nutrients and Energy in Fast Growing Meat-Type Chickens

Two consecutive growth studies were conducted to investigate the age-dependent nutrient and energy deposition in male and female meat-type chickens (Ross 308) based on feather and feather-free body fraction data determined according to the comparative slaughter technique. Birds were reared under standardized housing conditions (15 floor pens per gender, 5 birds per pen). Both the starter (day 1 to 22) and grower diets (day 22 to 36) were based on corn, wheat, soybean meal, soybean protein concentrate and crystalline feed amino acids. Diets were formulated to ensure an equal feed protein quality close to the ideal amino acid ratio by adjusting a constant mixture of the feed proteins. Individual body weight (BW) and feed intake per pen were recorded weekly. At the start (day 1) as well as on a weekly basis until the end of the 5th week, 15 birds per gender (each 3 pens of 5 birds) were selected and euthanized following 24 h feed deprivation. Subsequently, the feathers were manually removed and quantified. Crude nutrient analysed in representative samples of both feather and feather-free body fractions. The nutrient and energy deposition in the bodies of both genders were significantly increased with increasing age (p . Male birds deposited significantly higher body protein (p and female birds significantly more fat and energy in the whole body (p during the entire growth period. In contrast, no differences were found in protein contents of the BW gain between genders dependent on age (p > 0.05). However, the protein partitioning in the gain of both analyzed body fractions provided oppositional results. Accordingly, male birds yielded relatively more protein in the feather-free body fraction (p and females relatively more feather protein (p as related to the whole body protein gain.

Age and Gender Depending Growth of Feathers and Feather-Free Body in Modern Fast Growing Meat-Type Chickens

Two growth experiments with fast growing meat type chickens (Ross 308) were conducted to assess the growth of feathers and feather-free body dependent on age and gender (male:female ratio = 1:1). Birds were reared under uniform management and feeding conditions (floor pens;15 pens per gender;5 birds per pen) during the starter (day 1 to 22) and grower period (day 22 to 36). Diets were based on corn, wheat, soybean meal, soybean protein concentrate and balanced with feed amino acids to ensure an equal feed protein quality close to the ideal amino acid ratio by a constant mixture of the feed proteins. At start of the experiment and further on weekly up to the end of the 5th week, 15 birds per gender (each 3 pens of 5 birds) were selected and 24 h fasted before quantitative de-feathering. Both feather and feather free body fractions were significantly increased with increasing age of the birds (p < 0.001). Feather percentage as related to the empty body weight increased non-linearly from approximately 2% at the end of the first week to about 4% at the end of the experiment. Feather percentage and dry matter content of the feather-free body was significantly higher in female birds (p < 0.001) as compared to males. Further investigations will show how this varying proportions impact on nutrient deposition of modern meat-type chickens dependent on age and gender, respectively.

The Graded Inclusion of Algae (<i>Spirulina platensis</i>) or Insect (<i>Hermetia illucens</i>) Meal as a Soybean Meal Substitute in Meat Type Chicken Diets Impacts on Growth, Nutrient Deposition and Dietary Protein Quality Depending on the Extent of Amino Acid Supplementation

Three consecutive growth experiments were conducted to evaluate growth performance, whole body analyses and protein quality parameters from chicken diets with 50%, 75% and 100% substitution of soybean meal (SBM) by Spirulina meal (SM) or partly defatted Hermetia meal (HM). Each of the experiments was divided into a starter period (1 - 21 d) and a grower period (22 - 34 d). One-day-old male growing chickens (Ross 308) were randomly allotted to 48 floor pens making use of 6 birds/pen (Exp.1) or 7 birds/pen (Exp. 2, 3), and the experiments included a control diet (n = 12) and four experimental diets (n = 9). Experiment 1 examined a 50 % replacement of SBM by the alternative proteins under study, both on a basic and an advanced level of amino acid (AA) fortification to meet the recommended ideal amino acid ratio (IAAR). In experiment 2, 75% (starter diet) and 50% (grower diet) replacement of SBM was investigated. Experiment 3 investigated the effects of complete SBM substitution by SM or HM in starter and grower diets. In the second and third experiment diets with both of the alternative proteins and the control diet were AA supplemented to meet the current IAAR. In a further step, the calculated first limiting AA (LAA) was reduced to 80% of its requirement recommendation to allow for further evaluation of the individual AA efficiency according to the “Goettingen approach”. Different levels (50%, 75%, or 100%) of replacing SBM by HM or SM in chicken diets depressed dietary protein quality (p 0.001) and zoo-technical parameters (p 0.001) with only a basic level of AA supplementation. This effect was much more pronounced in diets with SM. However, with an extended level of dietary AA supplementation zoo-technical parameters (p 0.001), crude protein deposition (p 0.001) and protein quality parameters (p 0.001) were significantly improved. HM diets with an advanced level of AA tended to provide higher dietary protein quality and growth performance as compared to all SBM replacement levels through SM with different levels of AA supplementation and control diets.