Selective genotyping strategies for a sib test scheme of a broiler breeder program.

BACKGROUND: In broiler breeding, genotype-by-environment interaction is known to result in a genetic correlation between body weight measured in bio-secure and commercial environments that is substantially less than 1. Thus, measuring body weights on sibs of selection candidates in a commercial environment and genotyping them could increase genetic progress. Using real data, the aim of this study was to evaluate which genotyping strategy and which proportion of sibs placed in the commercial environment should be genotyped to optimize a sib-testing breeding program in broilers. Phenotypic body weight and genomic information were collected on all sibs raised in a commercial environment, which allowed to retrospectively analyze different sampling strategies and genotyping proportions. RESULTS: Accuracies of genomic estimated breeding values (GEBV) obtained with the different genotyping strategies were assessed by computing their correlation with GEBV obtained when all sibs in the commercial environment were genotyped. Results showed that, compared to random sampling (RND), genotyping sibs with extreme phenotypes (EXT) resulted in higher GEBV accuracy across all genotyping proportions, especially for genotyping proportions of 12.5% or 25%, which resulted in correlations of 0.91 vs 0.88 for 12.5% and 0.94 vs 0.91 for 25% genotyped. Including pedigree on birds with phenotype in the commercial environment that were not genotyped increased accuracy at lower genotyping proportions, especially for the RND strategy (correlations of 0.88 vs 0.65 at 12.5% and 0.91 vs 0.80 at 25%), and a smaller but still substantial increase in accuracy for the EXT strategy (0.91 vs 0.79 for 12.5% and 0.94 vs 0.88 for 25% genotyped). Dispersion bias was virtually absent for RND if 25% or more birds were genotyped. However, GEBV were considerably inflated for EXT, especially when the proportion genotyped was low, which was further exacerbated if the pedigree of non-genotyped sibs was excluded. CONCLUSIONS: When less than 75% of all animals placed in a commercial environment are genotyped, it is recommended to use the EXT strategy, because it yields the highest accuracy. However, caution should be taken when interpreting the resulting GEBV because they will be over-dispersed. When 75% or more of the animals are genotyped, random sampling is recommended because it yields virtually no bias of GEBV and results in similar accuracies as the EXT strategy.

Validation with single-step SNPBLUP shows that evaluations can continue using a single mean of genotyped individuals, even with multiple breeds.

BACKGROUND: In genomic prediction, it is common to centre the genotypes of single nucleotide polymorphisms based on the allele frequencies in the current population, rather than those in the base generation. The mean breeding value of non-genotyped animals is conditional on the mean performance of genotyped relatives, but can be corrected by fitting the mean performance of genotyped individuals as a fixed regression. The associated covariate vector has been referred to as a 'J-factor', which if fitted as a fixed effect can improve the accuracy and dispersion bias of sire genomic estimated breeding values (GEBV). To date, this has only been performed on populations with a single breed. Here, we investigated whether there was any benefit in fitting a separate J-factor for each breed in a three-way crossbred population, and in using pedigree-based expected or genome-based estimated breed fractions to define the J-factors. RESULTS: For body weight at 7 days, dispersion bias decreased when fitting multiple J-factors, but only with a low proportion of genotyped individuals with selective genotyping. On average, the mean regression coefficients of validation records on those of GEBV increased with one J-factor compared to none, and further increased with multiple J-factors. However, for body weight at 35 days this was not observed. The accuracy of GEBV remained unchanged regardless of the J-factor method used. Differences between the J-factor methods were limited with correlations approaching 1 for the estimated covariate vector, the estimated coefficients of the regression on the J-factors, and the GEBV. CONCLUSIONS: Based on our results and in the particular design analysed here, i.e. all the animals with phenotype are of the same type of crossbreds, fitting a single J-factor should be sufficient, to reduce dispersion bias. Fitting multiple J-factors may reduce dispersion bias further but this depends on the trait and genotyping rate. For the crossbred population analysed, fitting multiple J-factors has no adverse consequences and if this is done, it does not matter if the breed fractions used are based on the pedigree-expectation or the genomic estimates. Finally, when GEBV are estimated from crossbred data, any observed bias can potentially be reduced by including a straightforward regression on actual breed proportions.

Classifying aneuploidy in genotype intensity data using deep learning.

Aneuploidy is the loss or gain of one or more chromosomes. Although it is a rare phenomenon in liveborn individuals, it is observed in livestock breeding populations. These breeding populations are often routinely genotyped and the genotype intensity data from single nucleotide polymorphism (SNP) arrays can be exploited to identify aneuploidy cases. This identification is a time-consuming and costly task, because it is often performed by visual inspection of the data per chromosome, usually done in plots of the intensity data by an expert. Therefore, we wanted to explore the feasibility of automated image classification to replace (part of) the visual detection procedure for any diploid species. The aim of this study was to develop a deep learning Convolutional Neural Network (CNN) classification model based on chromosome level plots of SNP array intensity data that can classify the images into disomic, monosomic and trisomic cases. A multispecies dataset enriched for aneuploidy cases was collected containing genotype intensity data of 3321 disomic, 1759 monosomic and 164 trisomic chromosomes. The final CNN model had an accuracy of 99.9%, overall precision was 1, recall was 0.98 and the F1 score was 0.99 for classifying images from intensity data. The high precision assures that cases detected are most likely true cases, however, some trisomy cases may be missed (the recall of the class trisomic was 0.94). This supervised CNN model performed much better than an unsupervised k-means clustering, which reached an accuracy of 0.73 and had especially difficult to classify trisomic cases correctly. The developed CNN classification model provides high accuracy to classify aneuploidy cases based on images of plotted X and Y genotype intensity values. The classification model can be used as a tool for routine screening in large diploid populations that are genotyped to get a better understanding of the incidence and inheritance, and in addition, avoid anomalies in breeding candidates.

Estimation of dam line composition of 3-way crossbred animals using genomic information.

BACKGROUND: In genomic prediction including data of 3- or 4-way crossbred animals, line composition is usually fitted as a regression on expected line proportions, which are 0.5, 0.25 and 0.25, respectively, for 3-way crossbred animals. However, actual line proportions for the dam lines can vary between ~ 0.1 and 0.4, and ignoring this variation may affect the genomic estimated breeding values of purebred selection candidates. Our aim was to validate a proposed gold standard to evaluate different approaches for estimating line proportions using simulated data, and to subsequently use this in actual 3-way crossbred broiler data to evaluate several other methods. RESULTS: Analysis of simulated data confirmed that line proportions computed from assigned breed-origin-of-alleles (BOA) provide a very accurate gold standard, even if the parental lines are closely related. Alternative investigated methods were linear regression of genotypes on line-specific allele frequencies, maximum likelihood estimation using the program ADMIXTURE, and the genomic relationship of crossbred animals with their maternal grandparents. The results from the simulated data showed that the genomic relationship with the maternal grandparent was most accurate, and least affected by closer relationships between the dam lines. Linear regression and ADMIXTURE performed similarly for unrelated lines, but their accuracy dropped considerably when the dam lines were more closely related. In almost all cases, estimates improved after adjusting them to ensure that the sum of dam line contributions within animals was equal to 0.5, and within dam line and across animals the average was equal to 0.25. Results from the broiler data were much more similar between methods. In both cases, stringent linkage disequilibrium pruning of genotype data led to a relatively low accuracy of predicted line proportions, due to the loss of too many single nucleotide polymorphisms. CONCLUSIONS: With relatively unrelated parental lines as typical in crosses in pigs and poultry, linear regression of crossbred genotypes on line-specific allele frequencies and ADMIXTURE are very competitive methods. Thus, linear regression may be the method of choice, as it does not require genotypes of grandparents, is computationally very efficient, and easily implemented and adapted for considering the specific nature of the crossbred animals analysed.

Avian Primordial Germ Cells Are Bipotent for Male or Female Gametogenesis.

In birds, males are the homogametic sex (ZZ) and females are the heterogametic sex (ZW). Here, we investigate the role of chromosomal sex and germ cell competition on avian germ cell differentiation. We recently developed genetically sterile layer cockerels and hens for use as surrogate hosts for primordial germ cell (PGC) transplantation. Using in vitro propagated and cryopreserved PGCs from a pedigree Silkie broiler breed, we now demonstrate that sterile surrogate layer hosts injected with same sex PGCs have normal fertility and produced pure breed Silkie broiler offspring when directly mated to each other in Sire Dam Surrogate mating. We found that female sterile hosts carrying chromosomally male (ZZ) PGCs formed functional oocytes and eggs, which gave rise to 100% male offspring after fertilization. Unexpectedly, we also observed that chromosomally female (ZW) PGCs carried by male sterile hosts formed functional spermatozoa and produced viable offspring. These findings demonstrate that avian PGCs are not sexually restricted for functional gamete formation and provide new insights for the cryopreservation of poultry and other bird species using diploid stage germ cells.

Effect of genomic selection and genotyping strategy on estimation of variance components in animal models using different relationship matrices.

BACKGROUND: The traditional way to estimate variance components (VC) is based on the animal model using a pedigree-based relationship matrix (A) (A-AM). After genomic selection was introduced into breeding programs, it was anticipated that VC estimates from A-AM would be biased because the effect of selection based on genomic information is not captured. The single-step method (H-AM), which uses an H matrix as (co)variance matrix, can be used as an alternative to estimate VC. Here, we compared VC estimates from A-AM and H-AM and investigated the effect of genomic selection, genotyping strategy and genotyping proportion on the estimation of VC from the two methods, by analyzing a dataset from a commercial broiler line and a simulated dataset that mimicked the broiler population. RESULTS: VC estimates from H-AM were severely overestimated with a high proportion of selective genotyping, and overestimation increased as proportion of genotyping increased in the analysis of both commercial and simulated data. This bias in H-AM estimates arises when selective genotyping is used to construct the H-matrix, regardless of whether selective genotyping is applied or not in the selection process. For simulated populations under genomic selection, estimates of genetic variance from A-AM were also significantly overestimated when the effect of genomic selection was strong. Our results suggest that VC estimates from H-AM under random genotyping have the expected values. Predicted breeding values from H-AM were inflated when VC estimates were biased, and inflation differed between genotyped and ungenotyped animals, which can lead to suboptimal selection decisions. CONCLUSIONS: We conclude that VC estimates from H-AM are biased with selective genotyping, but are close to expected values with random genotyping.VC estimates from A-AM in populations under genomic selection are also biased but to a much lesser degree. Therefore, we recommend the use of H-AM with random genotyping to estimate VC for populations under genomic selection. Our results indicate that it is still possible to use selective genotyping in selection, but then VC estimation should avoid the use of genotypes from one side only of the distribution of phenotypes. Hence, a dual genotyping strategy may be needed to address both selection and VC estimation.

Association analysis of loci implied in "buffering" epistasis.

The existence of buffering mechanisms is an emerging property of biological networks, and this results in the buildup of robustness through evolution. So far, there are no explicit methods to find loci implied in buffering mechanisms. However, buffering can be seen as interaction with genetic background. Here we develop this idea into a tractable model for quantitative genetics, in which the buffering effect of one locus with many other loci is condensed into a single statistical effect, multiplicative on the total additive genetic effect. This allows easier interpretation of the results and simplifies the problem of detecting epistasis from quadratic to linear in the number of loci. Using this formulation, we construct a linear model for genome-wide association studies that estimates and declares the significance of multiplicative epistatic effects at single loci. The model has the form of a variance components, norm reaction model and likelihood ratio tests are used for significance. This model is a generalization and explanation of previous ones. We test our model using bovine data: Brahman and Tropical Composite animals, phenotyped for body weight at yearling and genotyped at high density. After association analysis, we find a number of loci with buffering action in one, the other, or both breeds; these loci do not have a significant statistical additive effect. Most of these loci have been reported in previous studies, either with an additive effect or as footprints of selection. We identify buffering epistatic SNPs present in or near genes reported in the context of signatures of selection in multi-breed cattle population studies. Prominent among these genes are those associated with fertility (INHBA, TSHR, ESRRG, PRLR, and PPARG), growth (MSTN, GHR), coat characteristics (KIT, MITF, PRLR), and heat resistance (HSPA6 and HSPA1A). In these populations, we found loci that have a nonsignificant statistical additive effect but a significant epistatic effect. We argue that the discovery and study of loci associated with buffering effects allow attacking the difficult problems, among others, of the release of maintenance variance in artificial and natural selection, of quick adaptation to the environment, and of opposite signs of marker effects in different backgrounds. We conclude that our method and our results generate promising new perspectives for research in evolutionary and quantitative genetics based on the study of loci that buffer effect of other loci.

Genetic analysis on body weight at different ages in broiler chicken raised in commercial environment.

A multivariate model was developed and used to estimate genetic parameters of body weight (BW) at 1-6 weeks of age of broilers raised in a commercial environment. The development of model was based on the predictive ability of breeding values evaluated from a cross-validation procedure that relied on half-sib correlation. The multivariate model accounted for heterogeneous variances between sexes through standardization applied to male and female BWs differently. It was found that the direct additive genetic, permanent environmental maternal and residual variances for BW increased drastically as broilers aged. The drastic increase in variances over weeks of age was mainly due to scaling effects. The ratio of the permanent environmental maternal variance to phenotypic variance decreased gradually with increasing age. Heritability of BW traits ranged from 0.28 to 0.33 at different weeks of age. The direct genetic effects on consecutive weekly BWs had high genetic correlations (0.85-0.99), but the genetic correlations between early and late BWs were low (0.32-0.57). The difference in variance components between sexes increased with increasing age. In conclusion, the permanent environmental maternal effect on broiler chicken BW decreased with increasing age from weeks 1 to 6. Potential bias of the model that considered identical variances for sexes could be reduced when heterogeneous variances between sexes are accounted for in the model.

Correction to: Validation of genomic predictions for body weight in broilers using crossbred information and considering breed-of-origin of alleles.

Following publication of original article [1], we noticed that there was an error: Eq. (3) on page 5 is the genomic relationship matrix that.

Correction to: Use of genomic information to exploit genotype-by-environment interactions for body weight of broiler chicken in bio-secure and production environments.

After publication of this work [1], we noticed that there was an error: the formula to calculate the standard error of the estimated correlation.

Genetic parameters for body weight and different definitions of residual feed intake in broiler chickens.

BACKGROUND: The objectives of this study were to (1) simultaneously estimate genetic parameters for BW, feed intake (FI), and body weight gain (Gain) during a FI test in broiler chickens using multi-trait Bayesian analysis; (2) derive phenotypic and genetic residual feed intake (RFI) and estimate genetic parameters of the resulting traits; and (3) compute a Bayesian measure of direct and correlated superiority of a group selected on phenotypic or genetic residual feed intake. A total of 56,649 male and female broiler chickens were measured at one of two ages ([Formula: see text] or [Formula: see text] days). BW, FI, and Gain of males and females at the two ages were considered as separate traits, resulting in a 12-trait model. Phenotypic RFI ([Formula: see text]) and genetic RFI ([Formula: see text]) were estimated from a conditional distribution of FI given BW and Gain using partial phenotypic and partial genetic regression coefficients, respectively. RESULTS: Posterior means of heritability for BW, FI and Gain were moderately high and estimates were significantly different between males and females at the same age for all traits. In addition, the genetic correlations between male and female traits at the same age were significantly different from 1, which suggests a sex-by-genotype interaction. Genetic correlations between [Formula: see text] and [Formula: see text] were significantly different from 1 at an older age but not at a younger age. CONCLUSIONS: The results of the multivariate Bayesian analyses in this study showed that genetic evaluation for production and feed efficiency traits should take sex and age differences into account to increase accuracy of selection and genetic gain. Moreover, for communicating with stakeholders, it is easier to explain results from selection on [Formula: see text] than selection on [Formula: see text], since [Formula: see text] is genetically independent of production traits and it explains the efficiency of birds in nutrient utilization independently of energy requirements for production and maintenance.

Use of genomic information to exploit genotype-by-environment interactions for body weight of broiler chicken in bio-secure and production environments.

BACKGROUND: The increase in accuracy of prediction by using genomic information has been well-documented. However, benefits of the use of genomic information and methodology for genetic evaluations are missing when genotype-by-environment interactions (G × E) exist between bio-secure breeding (B) environments and commercial production (C) environments. In this study, we explored (1) G × E interactions for broiler body weight (BW) at weeks 5 and 6, and (2) the benefits of using genomic information for prediction of BW traits when selection candidates were raised and tested in a B environment and close relatives were tested in a C environment. METHODS: A pedigree-based best linear unbiased prediction (BLUP) multivariate model was used to estimate variance components and predict breeding values (EBV) of BW traits at weeks 5 and 6 measured in B and C environments. A single-step genomic BLUP (ssGBLUP) model that combined pedigree and genomic information was used to predict EBV. Cross-validations were based on correlation, mean difference and regression slope statistics for EBV that were estimated from full and reduced datasets. These statistics are indicators of population accuracy, bias and dispersion of prediction for EBV of traits measured in B and C environments. Validation animals were genotyped and non-genotyped birds in the B environment only. RESULTS: Several indications of G × E interactions due to environmental differences were found for BW traits including significant re-ranking, heterogeneous variances and different heritabilities for BW measured in environments B and C. The genetic correlations between BW traits measured in environments B and C ranged from 0.48 to 0.54. The use of combined pedigree and genomic information increased population accuracy of EBV, and reduced bias of EBV prediction for genotyped birds compared to the use of pedigree information only. A slight increase in accuracy of EBV was also observed for non-genotyped birds, but the bias of EBV prediction increased for non-genotyped birds. CONCLUSIONS: The G × E interaction was strong for BW traits of broilers measured in environments B and C. The use of combined pedigree and genomic information increased population accuracy of EBV substantially for genotyped birds in the B environment compared to the use of pedigree information only.

Assessment of sire contribution and breed-of-origin of alleles in a three-way crossbred broiler dataset.

Broiler breeding programs rely on crossbreeding. With genomic selection, widespread use of crossbred performance in breeding programs comes within reach. Commercial crossbreds, however, may have unknown pedigrees and their genomes may include DNA from 2 to 4 different breeds. Our aim was, for a broiler dataset with a limited number of sires having both purebred and crossbred offspring generated using natural mating, to rapidly derive parentage, assess the distribution of the sire contribution to the offspring generation, and to assess breed-of-origin of alleles in crossbreds. The dataset contained genotypes for 56,075 SNPs for 5,882 purebred and 10,943 3-way crossbred offspring generated by natural mating of 164 purebred sires to 1,016 purebred and 1,386 F1 crossbred hens. Using our algorithm FindParents, joint parentage derivation for the offspring and parent generations required only 1 m 29 s to retrieve parentage for 20,253 animals considering 4,504 possible parents. FindParents was similarly accurate as a maximum likelihood based method, apart from situations where settings of FindParents did not match the genotyping error rate in the data. Numbers of offspring per sire had a very skewed distribution, ranging from 1 to 270 crossbreds and 1 to 154 purebreds. Derivation of breed-of-origin of alleles relied on phasing all genotypes, including 8,205, 372, and 720 animals from the 3 pure lines involved, and allocating haplotypes in the crossbreds to purebred lines based on observed frequencies in the purebred lines. Breed-of-origin could be derived for 96.94% of the alleles of the 1,386 F1 crossbred hens and for 91.88% of the alleles of the 10,943 3-way crossbred offspring, of which 49.49% to the sire line. The achieved percentage of assignment to the sire line was sufficient to proceed with subsequent analyses requiring only the breed-of-origin of the paternal alleles to be known. Although required number of animals may be population dependent, to increase the total percentage of assigned alleles, it seems advisable to use at least approx. 1,000 genotyped purebred animals for each of the lines involved.

Validation of genomic predictions for body weight in broilers using crossbred information and considering breed-of-origin of alleles.

BACKGROUND: Pig and poultry breeding programs aim at improving crossbred (CB) performance. Selection response may be suboptimal if only purebred (PB) performance is used to compute genomic estimated breeding values (GEBV) because the genetic correlation between PB and CB performance ([Formula: see text]) is often lower than 1. Thus, it may be beneficial to use information on both PB and CB performance. In addition, the accuracy of GEBV of PB animals for CB performance may improve when the breed-of-origin of alleles (BOA) is considered in the genomic relationship matrix (GRM). Thus, our aim was to compare scenarios where GEBV are computed and validated by using (1) either CB offspring averages or individual CB records for validation, (2) either a PB or CB reference population, and (3) a GRM that either accounts for or ignores BOA in the CB individuals. For this purpose, we used data on body weight measured at around 7 (BW7) or 35 (BW35) days in PB and CB broiler chickens and evaluated the accuracy of GEBV based on the correlation GEBV with phenotypes in the validation population (validation correlation). RESULTS: With validation on CB offspring averages, the validation correlation of GEBV of PB animals for CB performance was lower with a CB reference population than with a PB reference population for BW35 ([Formula: see text] = 0.96), and about equal for BW7 ([Formula: see text] = 0.80) when BOA was ignored. However, with validation on individual CB records, the validation correlation was higher with a CB reference population for both traits. The use of a GRM that took BOA into account increased the validation correlation for BW7 but reduced it for BW35. CONCLUSIONS: We argue that the benefit of using a CB reference population for genomic prediction of PB animals for CB performance should be assessed either by validation on CB offspring averages, or by validation on individual CB records while using a GRM that accounts for BOA in the CB individuals. With this recommendation in mind, our results show that the accuracy of GEBV of PB animals for CB performance was equal to or higher with a CB reference population than with a PB reference population for a trait with an [Formula: see text] of 0.8, but lower for a trait with an [Formula: see text] of 0.96. In addition, taking BOA into account was beneficial for a trait with an [Formula: see text] of 0.8 but not for a trait with an [Formula: see text] of 0.96.

Estimating the purebred-crossbred genetic correlation of body weight in broiler chickens with pedigree or genomic relationships.

BACKGROUND: In pig and poultry breeding programs, the breeding goal is to improve crossbred (CB) performance, whereas selection in the purebred (PB) lines is often based on PB performance. Thus, response to selection may be suboptimal, because the genetic correlation between PB and CB performance ([Formula: see text]) is generally lower than 1. Accurate estimates of the [Formula: see text] are needed, so that breeders can decide if they should collect data from CB animals. [Formula: see text] can be estimated either from pedigree or genomic relationships, which may produce different results. With genomic relationships, the [Formula: see text] estimate could be improved when relationships between purebred and crossbred animals are based only on the alleles that originate from the PB line of interest. This work presents the first comparison of estimated [Formula: see text] and variance components of body weight in broilers, using pedigree-based or genotype-based models, where the breed-of-origin of alleles was either ignored or considered. We used genotypes and body weight measurements of PB and CB animals that have a common sire line. RESULTS: Our results showed that the [Formula: see text] estimates depended on the relationship matrix used. Estimates were 5 to 25% larger with genotype-based models than with pedigree-based models. Moreover, [Formula: see text] estimates were similar (max. 7% difference) regardless of whether the model considered breed-of-origin of alleles or not. Standard errors of [Formula: see text] estimates were smaller with genotype-based than with pedigree-based methods, and smaller with models that ignored breed-of-origin than with models that considered breed-of-origin. CONCLUSIONS: We conclude that genotype-based models can be useful for estimating [Formula: see text], even when the PB and CB animals that have phenotypes are closely related. Considering breed-of-origin of alleles did not yield different estimates of [Formula: see text], probably because the parental breeds of the CB animals were distantly related.

Benefits of testing in both bio-secure and production environments in genomic selection breeding programs for commercial broiler chicken.

BACKGROUND: A breeding program for commercial broiler chicken that is carried out under strict biosecure conditions can show reduced genetic gain due to genotype by environment interactions (G × E) between bio-secure (B) and commercial production (C) environments. Accuracy of phenotype-based best linear unbiased prediction of breeding values of selection candidates using sib-testing in C is low. Genomic prediction based on dense genetic markers may improve accuracy of selection. Stochastic simulation was used to explore the benefits of genomic selection in breeding schemes for broiler chicken that include birds in both B and C for assessment of phenotype. RESULTS: When genetic correlations ([Formula: see text]) between traits measured in B and C were equal to 0.5 and 0.7, breeding schemes with 15, 30 and 45% of birds assessed in C resulted in higher genetic gain for performance in C compared to those without birds in C. The optimal proportion of birds phenotyped in C for genetic gain was 30%. When the proportion of birds in C was optimal and genotyping effort was limited, allocating 30% of the genotyping effort to birds in C was also the optimal genotyping strategy for genetic gain. When [Formula: see text] was equal to 0.9, genetic gain for performance in C was not improved with birds in C compared to schemes without birds in C. Increasing the heritability of traits assessed in C increased genetic gain significantly. Rates of inbreeding decreased when the proportion of birds in C increased because of a lower selection intensity among birds retained in B and a reduction in the probability of co-selecting close relatives. CONCLUSIONS: If G × E interactions ([Formula: see text] of 0.5 and 0.7) are strong, a genomic selection scheme in which 30% of the birds hatched are phenotyped in C has larger genetic gain for performance in C compared to phenotyping all birds in B. Rates of inbreeding decreased as the proportion of birds moved to C increased from 15 to 45%.

Estimating the genetic merit of sires by using pooled DNA from progeny of undetermined pedigree.

BACKGROUND: DNA-based predictions for hard-to-measure production traits hold great promise for selective breeding programs. DNA pooling might provide a cheap genomic approach to use phenotype data from commercial flocks which are commonly group-mated with parentage unknown. This study on sheep explores if genomic breeding values for stud sires can be estimated from genomic relationships that were obtained from pooled DNA in combination with phenotypes from commercial progeny. METHODS: Phenotypes used in this study were categorical data. Blood was pooled strategically aiming at even pool sizes and within sex and phenotype category. A hybrid genomic relationship matrix was constructed relating pools to sires. This matrix was used to determine the contribution of sires to each of the pools and therefore phenotype category by using a simple regression approach. Genomic breeding values were also estimated using the hybrid genomic relationship matrix. RESULTS: We demonstrated that, using pooled DNA, the genetic performance of sires can be illustrated as their contribution to phenotype categories and can be expressed as a regression coefficient. Genomic estimated breeding values for sires were equivalent to the regression coefficients and are a commonly used industry tool. CONCLUSIONS: Genotyping of DNA from pooled biological samples offers a cheap method to link phenotypic information from commercial production animals to the breeding population and can be turned into information on the genetic value of stud sires for traits that cannot be measured in the stud environment.

Study of the optimum haplotype length to build genomic relationship matrices.

BACKGROUND: As genomic data becomes more abundant, genomic prediction is more routinely used to estimate breeding values. In genomic prediction, the relationship matrix ([Formula: see text]), which is traditionally used in genetic evaluations is replaced by the genomic relationship matrix ([Formula: see text]). This paper considers alternative ways of building relationship matrices either using single markers or haplotypes of different lengths. We compared the prediction accuracies and log-likelihoods when using these alternative relationship matrices and the traditional [Formula: see text] matrix, for real and simulated data. METHODS: For real data, we built relationship matrices using 50k genotype data for a population of Brahman cattle to analyze three traits: scrotal circumference (SC), age at puberty (AGECL) and weight at first corpus luteum (WTCL). Haplotypes were phased with hsphase and imputed with BEAGLE. The relationship matrices were built using three methods based on haplotypes of different lengths. The log-likelihood was considered to define the optimum haplotype lengths for each trait and each haplotype-based relationship matrix. RESULTS: Based on simulated data, we showed that the inverse of [Formula: see text] matrix and the inverse of the haplotype relationship matrices for methods using one-single nucleotide polymorphism (SNP) phased haplotypes provided coefficients of determination (R2) close to 1, although the estimated genetic variances differed across methods. Using real data and multiple SNPs in the haplotype segments to build the relationship matrices provided better results than the [Formula: see text] matrix based on one-SNP haplotypes. However, the optimal haplotype length to achieve the highest log-likelihood depended on the method used and the trait. The optimal haplotype length (7 to 8 SNPs) was similar for SC and AGECL. One of the haplotype-based methods achieved the largest increase in log-likelihood for SC, i.e. from -1330 when using [Formula: see text] to -1325 when using haplotypes with eight SNPs. CONCLUSIONS: Building the relationship matrix by using haplotypes that comprise multiple SNPs will increase the accuracy of estimated breeding values. However, the optimum haplotype length that shows the correct relationship among individuals for each trait can be derived from the data.

Genomic correlation: harnessing the benefit of combining two unrelated populations for genomic selection.

BACKGROUND: The success of genomic selection in animal breeding hinges on the availability of a large reference population on which genomic-based predictions of additive genetic or breeding values are built. Here, we explore the benefit of combining two unrelated populations into a single reference population. METHODS: The datasets consisted of 1829 Brahman and 1973 Tropical Composite cattle with measurements on five phenotypes relevant to tropical adaptation and genotypes for 71,726 genome-wide single nucleotide polymorphisms (SNPs). The underlying genomic correlation for the same phenotype across the two breeds was explored on the basis of consistent linkage disequilibrium (LD) phase and marker effects in both breeds. RESULTS: The proportion of genetic variance explained by the entire set of SNPs ranged from 37.5 to 57.6 %. Estimated genomic correlations were drastically affected by the process used to select SNPs and went from near 0 to more than 0.80 for most traits when using the set of SNPs with significant effects and the same LD phase in the two breeds. We found that, by carefully selecting the subset of SNPs, the missing heritability can be largely recovered and accuracies in genomic predictions can be improved six-fold. However, the increases in accuracy might come at the expense of large biases. CONCLUSIONS: Our results offer hope for the effective implementation of genomic selection schemes in situations where the number of breeds is large, the sample size within any single breed is small and the breeding objective includes many phenotypes.

Machine learning approach for pooled DNA sample calibration.

BACKGROUND: Despite ongoing reduction in genotyping costs, genomic studies involving large numbers of species with low economic value (such as Black Tiger prawns) remain cost prohibitive. In this scenario DNA pooling is an attractive option to reduce genotyping costs. However, genotyping of pooled samples comprising DNA from many individuals is challenging due to the presence of errors that exceed the allele frequency quantisation size and therefore cannot be simply corrected by clustering techniques. The solution to the calibration problem is a correction to the allele frequency to mitigate errors incurred in the measurement process. We highlight the limitations of the existing calibration solutions such as the fact they impose assumptions on the variation between allele frequencies 0, 0.5, and 1.0, and address a limited set of error types. We propose a novel machine learning method to address the limitations identified. RESULTS: The approach is tested on SNPs genotyped with the Sequenom iPLEX platform and compared to existing state of the art calibration methods. The new method is capable of reducing the mean square error in allele frequency to half that achievable with existing approaches. Furthermore for the first time we demonstrate the importance of carefully considering the choice of training data when using calibration approaches built from pooled data. CONCLUSION: This paper demonstrates that improvements in pooled allele frequency estimates result if the genotyping platform is characterised at allele frequencies other than the homozygous and heterozygous cases. Techniques capable of incorporating such information are described along with aspects of implementation.