Simulated Comparison Between Different Scenarios of Modular Chicken Breeding Program using MoBPSweb v.1.6.62

Background: An optimal selective breeding program must balance ethical risks and operational costs without 24 necessarily compromising its effectiveness and aims. For this purpose, a modular breeding simulator was 25 used. Modular simulation acts as a preliminary evaluation instrument to predict the entire likelihood of 26 outputs from simulation sensitivity. Therefore, these computational requirements may need to be considered one of the operational cost components, especially for the digital integrated poultry industry.

BACKGROUND 60 The domestic needs for poultry-based protein sources such as eggs and meats rely heavily on the 61 outsourced products, despite being facilitated locally (Ferlito and Respatiadi 2018). This high dependency 62 may jeopardize domestic and global food security, as Hodson (2017) described. Few may realize that global 63 food security has its limits and can no longer be sustained by conventional means or methods. On the other 64 hand, a growing concern about constant threats from environmental crises and the ever-increasing human 65 population adds to the problem. Although it may seem insignificant, this might mark the beginning of global 66 food security collapse. Gama Ayam Research Team creates a pathway to solving poultry problems in 67 Indonesia. A conceived solution was to tap into the vast biodiversity of Indonesian indigenous chicken 68 breeds. An extensive repertoire of Indonesian indigenous chicken breeds (Nataamijaya 2010; Henuk and 69 Bakti 2018; Mahardhika et al. 2021) provides a unique and promising opportunity to be developed. 70 Developing Indonesian indigenous chicken breeds would help secure food sources domestically, therefore 71 hindering any external interventions or collateral effects caused by the global food crisis. 72 For years, studies about one Indonesian indigenous chicken breed, Pelung chicken, have been undergone 73 since its conception as the potential candidate of Indonesia-owned meat-type chicken breed by Daryono et al. 74 (2010). During those years, a wide array of research involving selective breeding programs has developed 75 significant key findings regarding its phenotypes and molecular characteristics (Retnoaji et  conclusion, it was evident that the breed can be further developed to serve as a more sustainable and long-79 term solution for supporting the local poultry sector. For most of the years, the study mainly revolves around 80 three fundamental aspects: effectiveness, reliability, and minimum ethical cost (Mahardhika et al. 2021). 81 However, the complexity of the selective breeding program could not be handled so easily. The reason was 82 the breeding scheme or genetic architecture and the requisite necessity to create a balance between 83 operational cost, ethical costs, and breeding gains. Thus, it requires a direct experimental or field study and 84 an analytical study, which mainly focuses on predicting the program's outcome. 85 Collecting data from either internal or other relevant studies outside the Gama Ayam Research Team can 86 be utilized as selective parameters. This library of the selective parameter could serve as a pre-emptive 87 measure to avoid unnecessary chicken sacrifice, unethical rearing, cost requirement, and error probability 88 (Mahardhika et al. 2021). An advanced selective breeding program could be achieved using algorithms to 89 compute these selective parameters into a modular design of a chicken breeding scheme with different 90 scenarios and selections. A preliminary evaluation could be produced without strict parameters and the 91 flexibility to modulate or alternate the breeding scheme or scenarios. This digitalized state would provide an 92 instrument to evaluate, compensate or anticipate any outcome of the selective breeding program with reliable 93 precision and accuracy. The Modular Breeding Program Simulator web version (MoBPSweb) written and 94 constructed by Pook and the team was used to facilitate this purpose. Categorized as stochastic simulation, 95 MoBPS used the R language and was created based on the flaws of its predecessors (Pook et al. 2020;Pook 96 et al. 2021). This study investigated the candidate for meat-type Pelung chicken breed produced from the 97 actual selective breeding program Gama Ayam Kambro (Kampong-Broiler). Different selection scenarios 98 were compared and analyzed using MoBPSweb v.1.6.62. Specific selective parameters were formulated and 99 tested against the breeding scheme according to Gama Ayam Kambro's research. This study also explores the 100 feature provided by MoBPSweb to perform specific economic projections. 101

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Settings and parameters 103 The hardware used in this research is an ASUS X450JF-x64 notebook with core processor Intel® Core™ 104 i7 repetition and phenotypic information record (phenotyping) as much as 50 times (generations). Broiler-117 Pelung selection index and residual genetic correlation are applied. The designed selection model scenario 118 consist of scenario_1 (Genomic_SD), scenario_2 (Breeding Value_SD), and scenario_3 (Phenotypic_SD). 119 Each one of the selection model scenarios is tested toward the Gama Ayam Kambro breeding scheme. 120 Genomic parameters were constructed using the Ensembl Map Affymetrix Chicken 600K SNPs Array 121 genomic database. The marker density used is 300K SNPs, with the time unit simulation is set to per week. 122 Additional settings like miraculix and parallel computational are activated. In the genomic selection, the 123 SIM_08 procedure used five core processor with a maximum memory capacity of 30 GB and max the 124 maximum duration of the computational process is 48 hours. The effects of three selection model scenarios 125 on the Gama Ayam Kambro selective breeding program are known according to their output in each chicken 126 nucleus, from the elders to their tillers. The output consists of accuracy, F coefficient, kinship, observed 127 phenotypes, and economic parameter projections related to Gama Ayam Kambro selective breeding program.

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The breeding scheme was designed using nodes (nucleus or cohort) and edges (breeding action) 132 represented by color-coded boxes and arrows in sequence ( Figure 1). Boxes of pink represent the chicken 133 group for the hen nucleus (♀) and light blue for the rooster nucleus (♂). Breeding action is represented by 134 red arrows (repeat), green arrows (selection), and orange arrows (reproduction

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A review of accuracy from three selection model scenarios in the Gama Ayam Kambro selective 151 breeding program with 50 generations are elaborated per chicken nucleus for each selection index, from the 152 elders to their litters (Figure 2). For the Broiler selection index, in the nucleus of selected ♀ BC500 and ♂ 153 PBH elderly chicken, the three selection model scenarios have the upper and lower accuracy limit of 95% 154 and 60%, respectively, with the upper and lower limit of accuracy extreme fluctuations in each generation. In 155 the ♀/♂ 1 st and 2 nd Kambro chicken nucleus, there is still fluctuation followed by an increase in the 156 accuracy's lower limit to 70%. Then, stability begins to be achieved in the ♀/♂ 3 rd and 4 th Kambro chicken 157 nucleus, followed by a decrease in the accuracy's upper limit to 80%. In the ♀/♂ 3 rd and 4 th Kambro chicken 158 nucleus, an extreme lower limit of accuracy by -20% and 0%, respectively. When the elder ♀/♂ PBH 159 chicken nucleus been reintroducing into breeding, it was observed in the ♀/♂ 5 th Kambro chicken nucleus 160 there is fluctuation followed by an increase in upper and lower limits of accuracy to 95% and 75%, 161 respectively. It can be concluded that for the Broiler selection index, there is an increase in the lower limit of 162 accuracy by 15% in the 5 th Kambro chicken nucleus against the nucleus of selected elders chicken, which 163 indicates a decrease in the range of accuracy fluctuations by 15 points. According to this discovery, it is 164 pretty practical to use an outbreeding technique in increasing the accuracy of genomic prediction of the three 165 selection model scenarios was known. Implicitly, these three selection model scenarios reveal the fluctuation 166 patterns uniformity from the accuracy result. The accuracy result (r50) over 50 generations shows that 167 scenario_1 is the most suitable for selecting ♂ 5 th Kambro chicken and ♂ Pelung elders chickens using 168 the Broiler selection index (Figure 2). 169 For the Pelung selection index, in the nucleus of the ♀ BC500 and ♂ PBH selected elder chickens, three 170 selection model scenarios have an accuracy limitation in upper and lower by 95% and 50%, in order with an 171 extreme fluctuation in each generation. In the ♀/♂ 1 st and 2 nd Kambro chicken nucleus, the fluctuation is still 172 happened, followed by the increase of accuracy's lower limit to 65% and 70%, respectively. In the ♀/♂ 3 rd 173 and 4 th Kambro chicken nucleus begin to achieve stability, followed by a decrease in the upper accuracy limit 174 to 80%. In addition, extreme lower accuracy limits of -20% and 0% were detected, respectively. When the 175 ♀/♂ PBH elders chicken nucleus being reintroduced in the breeding, it is observed that ♀/♂ 5 th Kambro 176 chicken nucleus fluctuations occur followed by an increase in the upper and lower limit of accuracy to 90% 177 and 70%, respectively. 178 To conclude, for the Pelung selection index, the upper limit of 5 th Kambro nucleus accuracy decreased 179 by 5% against the nucleus of selected elders chicken meanwhile, there is a 20% increase for the lower limit. 180 The fluctuation of the upper-and lower limit indicates a derivation in the range of accuracy by 25 points. 181 According to this discovery, using an outbreeding technique to increase genomic prediction accuracy from 182 the three selection model scenarios is pretty practical. In the entire perspective, these three selection model 183 scenarios show a uniformity of fluctuation pattern from obtaining accuracy. According to the acquisition of 184 accuracy (r50) of 50 generations, it is showing that scenario_1 is the most suitable for selecting ♂ 5 th Kambro 185 chicken and ♂ Pelung elders chickens using the Pelung selection index (Figure 2). 186 The decrease in the accuracy fluctuating range from the three selection model scenarios in the Pelung 187 selection index is more significant than Broiler. However, the lower and upper limits of the 5 th Kambro 188 chicken nucleus accuracy for the Pelung selection index are lessened than Broiler. On the other hand, in the 189 lower limit of elders, chicken nucleus accuracy for Broiler index selection are higher than Pelung. The 190 conclusion is that the use of the Broiler selection index is adequate than Pelung. The Pelung selection index 191 assigns a higher value to the FEML (5) and TL (5) phenotype characters, while the Broiler selection index to 192 FCR (5), BW49D (4), and BW56D (4). According to this discovery, weighting affects the accuracy of the

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An overview regarding the F coefficient and weekly kinship of the three selection model scenarios in the 205 Gama Ayam Kambro selective breeding program is described in each chicken nucleus, starting from the 206 elders to the litters (Figure 3). For the F coefficient, in the nucleus of selected ♀ BC500 and ♂ PBH elders, it 207 is known that three selection scenarios have upper and lower limits of 0.3 and 0, respectively, with a linear 208 increase pattern per week. In the ♀/♂ 1 st Kambro nucleus, an increasing linear pattern is still detected, 209 followed by a decrease in the upper limit of the F coefficient to 0.25. Meanwhile, the ♀/♂ 2 nd Kambro, an 210 increased linear pattern followed by an increase in the upper and lower limits to 0.42 and 0.26, respectively. 211 In the ♀/♂ 3 rd and 4 th Kambro nucleus, there is an F coefficient of scenario_1 with scenario_2 and 212 scenario_3. As a result, scenario_1 has a higher increasing pattern of the F coefficient than scenario_2 and scenario_3. In the 3 rd Kambro nucleus, an increase in upper and lower limits of the F coefficient was detected 214 to 0.52 and 0.4, respectively. Meanwhile, an increase in the 4 th Kambro nucleus was detected to 0.58 and 0.5, 215 respectively. As the ♀/♂ PBH elders nucleus was reintroduced to breeding, it was observed that the ♀/♂ 5 th 216 Kambro nucleus fluctuated and followed by a decrease in the upper and lower limits of the F coefficient to 217 0.035 and 0.005, respectively (Figure 3). 218 According to upper and lower limits, there is a weekly increase in the F coefficient from the 2 nd 219 Kambro to the 4 th Kambro nucleus. In summary, there is a decrease in the upper limit of the 5 th Kambro 220 nucleus F coefficient by 0.265 points against the nucleus of the selected elders, while for the lower limit, 221 there is an increase of 0.005 points. In addition, this also indicates a fluctuating range decrease in 222 the F coefficient by 0.04 points. According to these findings, using the outbreeding technique effectively 223 reduces the F coefficient of the three selection model scenarios. There is a separation of the F coefficient of 224 scenario_1 with scenario_2 and scenario_3 in the 3 rd and 4 th Kambro chicken nucleus. At least three factors 225 most likely underlie this occurrence: the choice of broodstock, crossing techniques, and the selected design. 226 It has been recognized that the F coefficient of the 2 nd Kambro's nucleus has reached the relatively high 227 relative limit to the nucleus of the elders or 1 st Kambro chicken. It is suspected to amplify the homozygotic 228 allele fraction when the selected 2 nd Kambro is crossed with each other based on the common inbreeding 229 technique to form ♀/♂ 3 rd Kambro nucleus. A similar procedure was also applied in forming the nucleus of 230 ♀/♂ 4 th Kambro. The selection model scenario factor is might play a passive role due to the accumulation of 231 the two previous factors. According to F coefficient (F5000) for 5000 weeks, it was found that scenario_1 with 232 the most practical selection design for selecting ♂ 5 th Kambro and elder ♂ Pelung chicken under the Gama 233 Ayam Kambro breeding scheme (Figure 3).

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In terms of kinship, in the nucleus of the selected ♀ BC500 ♂ PBH and elders, it has known that the 238 three selection model scenarios have upper and lower limits of 0.5 and 0, respectively, with a linear increase 239 pattern per week. In the ♀/♂ 1 st Kambro nucleus, the increasing linear pattern is still detected without 240 changing the kinship's upper and lower limits. In ♀/♂ 2 nd Kambro nucleus, a linear pattern was improved, 241 followed by an increase in the upper and lower to 0.8 and 0.55, respectively. In the ♀/♂ 3 rd and 4 th Kambro 242 nucleus, there is a separation of kinship between scenario_1 with scenario_2 and scenario_3. Scenario_1 has 243 a higher increasing kinship pattern than scenario_2 and scenario_3. In the 3 rd Kambro nucleus, there is an 244 uplift in the upper and lower limits of kinship to 1 and 0.8, respectively.
Meanwhile, an increase in the 4 th Kambro nucleus was detected to 1.15 and 1, respectively. When the 246 ♀/♂ PBH elders being reintroduced into breeding, it observed there is a fluctuation occurred and followed by 247 a decrease in the upper and lower limits of kinship to 0.3 and 0 in the ♀/♂ 5 th Kambro nucleus, respectively 248 ( Figure 3). According to upper and lower limits, there is an increase in the weekly kinship starting from the 249 2 nd Kambro nucleus to the 4 th Kambro. Therefore, it can be assumed that there is a decrease in the 5 th Kambro 250 kinship's upper limit by 0.2 points to the chosen elders chicken's nucleus. According to this discovery, using 251 outbreeding techniques is quite effective in reducing the homozygotic allele fraction of the three selection 252 model scenarios. The kinship strengthens the factors of broodstock and crossing technique as 253 the F coefficient separation of in the 3 rd and 4 th Kambro. In addition, it also proves the amplification of the 254 homozygotic allele fraction, which leads to an increase in the F coefficient. It is concluded that the cohort's 255 close kinship increases the F coefficient. According to the kinship (R5000) over 5000 weeks, it was found that 256 scenario_1 with the genomic selection design is the most applicable for selecting ♂ 5 th Kambro and elders of 257 ♂ Pelung chicken, under the Gama Ayam Kambro chicken breeding scheme (Figure 3).

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An overview of the observed phenotypes of AFE from the three selection model scenarios in the Gama 262 Ayam Kambro selective breeding program with 50 generations was described per nucleus, from the elders to 263 litters (Figure 4). In the nucleus of selected ♀ BC500 elders and ♂ PBH, the three selection model scenarios 264 have upper and lower limits of AFE by 169 and 161 days, in order with a linear increase pattern per 265 generation. The lowest AFE was detected in the same nucleus at scenario_2, while the highest is in 266 scenario_3, followed by scenario_1. In the ♀/♂ 1 st Kambro nucleus, linear increases pattern was still 267 detected, followed by a decrease in the upper limit of AFE to 168 days. While in the ♀/♂ 2 nd Kambro 268 nucleus, a linear increases pattern followed by an increase in the upper and lower limits to 171 and 166 days, 269 respectively. The ♀/♂ 3 rd Kambro nucleus, an increase in the upper and lower limits of AFE was detecter to 270 174 and 169 days, respectively. While in the ♀/♂ 4 th Kambro nucleus, an increase was detected to 175 and 271 171 days, respectively. When the elders ♀/♂ PBH nucleus being reintroduced to breeding, it was observed in 272 the ♀/♂ 5 th Kambro nucleus there are a decrease in the upper and lower limits of AFE to 174 and 160days, 273 respectively ( Figure 4). According to upper and lower limits, there is an increase in AFE starting from the 274 nucleus of 2 nd Kambro to 4 th Kambro. There is an increase in the upper limit of AFE 5 th Kambro nucleus by 5 275 points to the nucleus of the selected elders, while the lower limit decreased by 1 point. Moreover, that 276 indicates an increase in AFE fluctuating range by 6 points. According to these findings, using the 277 outbreeding technique reduces AFE from the three selection model scenarios. Based on AFE (AFE50) 278 throughout 50 generations, it is discovered that scenario_2 (BV) dan scenario_3 (PHEN) is the most 279 applicable for selecting the ♂ 5 th Kambro and elder ♂ Pelung, respectively under the Gama Ayam Kambro 280 breeding scheme (Figure 4).

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An overview of an observed phenotype of BW49D from the three selection model scenarios in the Gama 285 Ayam Kambro selective breeding program with 50 generations was described per nucleus, starting from the 286 elders to their litters ( Figure 5). In the nucleus of selected elders ♀ BC500 and ♂ PBH, it has known that the 287 three selection model scenarios have upper and lower limits of BW49D of 780 and 710 grams, in order with 288 fluctuations per generation. In the same chicken nucleus, the lowest BW49D was detected in scenario_3, 289 while the highest was in scenario_2 and followed by scenario_1. The fluctuation in the nucleus of ♀/♂ 1 st 290 Kambro still happens, followed by a decrease in the upper limit and an increase in the BW49D's lower limit 291 to 770 and 720 grams, respectively. Meanwhile, in the ♀/♂ 2 nd Kambro nucleus, an increase in the linear 292 pattern was detected attenuation of fluctuations without changes in the upper and lower limits of BW49D 293 relatively to the 1 st Kambro nucleus. Finally, in the ♀/♂ 3 rd Kambro nucleus, it is indicated that there is an 294 increase in the upper limit and a decreased lower limit of BW49D to 780 and 710 grams, respectively. 295 Meanwhile, in the ♀/♂ 4 th Kambro nucleus, a decrease in the upper limit to 770 grams was detected. As 296 the nucleus of ♀/♂ PBH elders has been reintroducing into breeding, it has detected there was no change in 297 the upper and lower limits of BW49D in the nucleus of ♀/♂ 5 th Kambro. In the nucleus of ♀/♂ 5 th Kambro, 298 fluctuations, and convergence of the upper and lower limits of BW49D were detected from the three 299 selection model scenarios ( Figure 5). According to upper and lower limits, there are no changes in the 300 BW49D from the 1 st Kambro nucleus to the 4 th Kambro. It summarizes a decrease in the BW49D upper limit 301 of the 5 th Kambro nucleus by 10 points to the nucleus selected elders, while there was no change for the 302 lower limit. Otherwise, it indicates a decrease in the fluctuating range of BW49D by 10 points. In the 5 th 303 Kambro nucleus, there was a convergence of upper and lower limits in the range of 750 and 710 grams, 304 respectively. According to these findings, outbreeding techniques are less effective in increasing BW49D 305 from the three selection model scenarios. Based on BW49D (BW49D50) over 50 generations, found that 306 scenario_3 (PHEN) and scenario_2 (BV) are the most suitable for selecting ♂ 5 th Kambro chickens and ♂ 307 Pelung elder chickens, respectively under the Gama Ayam Kambro breeding scheme ( Figure 5). 308

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An overview of an observed phenotype of BW56D from the three selection model scenarios in the Gama 312 Ayam Kambro selective breeding program with 50 generations was described per nucleus, from the elders to 313 litters ( Figure 6). In the nucleus of selected ♀ BC500 and ♂ PBH elders, the three selection model scenarios respectively. In the nucleus of ♀/♂ 4 th Kambro, an increase is detected in the upper and lower limits to 2250 320 and 1950 grams, respectively. The lowest of BW56D in the same nucleus is in scenario_3, whereas the 321 highest is in scenario_1 and then scenario_2. When the nucleus of ♀/♂ PBH elders reintroduced into the 322 breeding, it was observed that there is a decrease in upper and lower limits of BW56D to 2200 and 900 323 grams in the ♀/♂ 5 th Kambro nucleus, respectively ( Figure 6). Based on upper and lower limits, there is an 324 increase of BW56D starting from the nucleus of 2 nd Kambro to 4 th Kambro. The conclusion is an increase in 325 the upper limit of BW56D in the 5 th Kambro nucleus by 700 points towards the elder's nucleus, whereas, for 326 the lower limit, there was a decrease of 100 points. Moreover, that indicates there is a decrease fluctuating 327 range by 200 points. According to these findings, the outbreeding technique effectively increases BW56D 328 from the three selection model scenarios. Based on BW56D (BW56D50) over 50 generations, scenario_1 329 (GEN) and scenario_3) are the most applicable for selecting ♂ 5 th Kambro chickens and ♂ Pelung chickens, 330 respectively under the Gama Ayam Kambro breeding scheme ( Figure 6). 331

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An overview of an observed phenotype of EN from the three selection model scenarios of the Gama 335 Ayam Kambro selective breeding program with 50 generations was described per nucleus, from the elders to 336 their litters (Figure 7). In the nucleus of selected ♀ BC500 and ♂ PBH elders, the three selection model 337 scenarios have an upper and lower limit of EN by 130 and 70 items/day, with fluctuations in each generation. 338 In the same nucleus, the lowest EN was detected in scenario_1, while the highest is in scenario_2, followed 339 by scenario_3. In the nucleus of ♀/♂ 1 st Kambro, still detect a fluctuation, followed by the upper limit of EN 340 decreasing to 120 items/day. Meanwhile, in the nucleus of ♀/♂ 2 nd Kambro, an increasing linear pattern with 341 a weakening fluctuation in the lower limit of EN to 60 items/day is detected. In the nucleus of ♀/♂ 3 rd dan 4 th 342 Kambro, an increasing linear pattern without any change on upper and lower limits of EN relatively to the 343 nucleus ♀/♂ 2 nd Kambro is detected. When the nucleus of ♀/♂ PBH elders reintroduced into the breeding, it 344 observed that in the nucleus of ♀/♂ 5 th Kambro, there is any change in upper and lower limits of EN ( Figure  345 7 breeding scheme (Figure 7). 352

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An overview of an observed phenotype of FCR from the three selection model scenarios of the Gama 356 Ayam Kambro selective breeding program with 50 generations was described per nucleus, from the elders to 357 their litter (Figure 8). In the nucleus of selected ♀ BC500 and ♂ PBH elders, the three selection model 358 scenarios have upper and lower limits of FCR by 4 and 2kg/kg, in order with fluctuations in each generation.

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In the same nucleus, the lowest FCR was detected in scenario_3, while the highest is in scenario_2, followed 360 by scenario_1. The nucleus of ♀/♂ 1 st Kambro has still detected fluctuation without any changes in the upper 361 and lower limits of FCR. Meanwhile, in the nucleus of ♀/♂ 2 nd Kambro, an increasing linear pattern with a 362 weakening fluctuation followed by an increase in the upper limit of FCR to 4,5 and 2,5kg/kg, respectively. In 363 the nucleus of ♀/♂ 3 rd dan 4 th Kambro, an increasing linear pattern is detected, followed by an increase of 364 upper and lower limits of FCR to 5 and 3kg/kg, respectively. When reintroduced the nucleus of ♀/♂ PBH 365 elders into the breeding, it observed that in the nucleus of ♀/♂ 5 th Kambro, there is a decrease in the lower 366 limit of FCR to 2kg/kg (Figure 8). Based on upper and lower limits, there is an increase in FCR that started 367 from the nucleus of 2 nd Kambro to 4 th Kambro. It can be concluded that there is an increase in the FCR upper 368 limit in the nucleus of 5 th Kambro by 1 point towards the nucleus of the elders, while there is no change for 369 the lower limit. In addition, it also indicates an increased fluctuating range of FCR by 1 point. According to 370 these findings, using the outbreeding technique is less effective in lowering the FCR of the three selection

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An overview of an observed phenotype of FEML from the three selection model scenarios of the Gama 378 Ayam Kambro selective breeding program with 50 generations was described per nucleus, from the elders to 379 their litters (Figure 9). In the nucleus of selected ♀ BC500 and ♂ PBH elders, the three selection model Kambro, an increasing linear pattern followed by increasing the upper and lower limits of FEML to 23 and 385 18 cm is detected, respectively. In the nucleus of ♀/♂ 3 rd Kambro, an increasing linear pattern was detected, 386 followed by increasing the upper and lower limits of FEML to 27 and 23 cm, respectively. Meanwhile, in the 387 nucleus of ♀/♂ 4 th Kambro, followed by increasing upper and lower limits of FEML by 28 and 25 cm, 388 respectively. When the nucleus of ♀/♂ PBH elders being reintroduced into the breeding, it observed a 389 decrease in the lower limits of FEML to 10 cm in the nucleus of ♀/♂ 5 th Kambro (Figure 9). Based on upper 390 and lower limits, an increasing FEML started from the nucleus 2 nd Kambro to 4 th Kambro. It summarizes an 391 increase in the upper limit of FEML in the nucleus of 5 th Kambro to 8 points towards the nucleus of elders, 392 which is a decrease by 2 points for the lower limit. In addition, it indicates an increase in the fluctuating 393 range of FEML by 10 points. Based on these findings, the outbreeding technique effectively increases the 394 FEML from the three selection model scenario. Based on FEML (FEML50) over 50 generations, scenario_2 395 (BV) and scenario_3 (PHEN) are the most suitable for selecting ♂ 5 th Kambro chickens and ♂ Pelung elder 396 chickens, respectively under the Gama Ayam Kambro breeding scheme (Figure 9). 397 Supplemental File 1. Parameters of MoBPSweb v.1.6.62