Screening for causative mutations in ovine BMPR1B and BMP15 genes and their homologous fragments in human.

PURPOSE: The BMPR1B and BMP15 genes are well known for their considerable associations with prolificacy in sheep. These genes may also affect fertility or prolificacy in other species, including human. This study was conducted to investigate possible causative mutations in BMPR1B and BMP15 genes in human and an indigenous breed of sheep. METHODS: Blood samples were collected from 83 singleton- and prolific Mehraban ewes and 81 infertile, singleton- and twin-bearing women. A 190-bp fragment, containing the FecB mutation in ovine BMPR1B, a 380-bp fragment in ovine BMP15 gene and their homologous fragments in human were amplified and then investigated by single-stranded conformation polymorphism and DNA sequencing methods. RESULTS: The FecB mutation of BMPR1B (g.159A>G) was detected in the sheep population, but no polymorphic loci were found in the homologous fragment in studied human samples. The studied fragments of BMP15 were monomorphic in both sheep and human samples. A total of nine and 69 point-differences in the studied fragments of BMPR1B and BMP15 genes were detected between the species, respectively. In sheep, the G allele of BMPR1B had a positive effect on litter size (p<0.05), whereby all AG or GG ewes were prolific. CONCLUSION: The FecB mutation for the first time was detected in Mehraban sheep and therefore could be considered for marker-assisted selection in this breed. The studied fragments of BMPR1B and BMP15 genes are not responsible for reproduction variation in human. More studies on other genes, associated with fertility in human, are necessary in the future.

Implementation of Feed Efficiency in Iranian Holstein Breeding Program.

This study aimed to evaluate the economic impact of improving feed efficiency on breeding objectives for Iranian Holsteins. Production and economic data from seven dairy herds were used to estimate the economic values of different traits, and a meta-analysis was conducted to analyze the genetic relationships between feed efficiency and other traits. Economic weights were calculated for various traits, with mean values per cow and per year across herds estimated at USD 0.34/kg for milk yield, USD 6.93/kg for fat yield, USD 5.53/kg for protein yield, USD -1.68/kg for dry matter intake, USD -1.70/kg for residual feed intake, USD 0.47/month for productive life, and USD -2.71/day for days open. The Iranian selection index was revised to improve feed efficiency, and the feed efficiency sub-index (FE$) introduced by the Holstein Association of the United States of America was adopted to reflect Iran's economic and production systems. However, there were discrepancies between Iranian and US genetic coefficients in the sub-index, which could be attributed to differences in genetic and phenotypic parameters, as well as the economic value of each trait. More accurate estimates of economic values for each trait in FE$ could be obtained by collecting dry matter intake from Iranian herds and conducting genetic evaluations for residual feed intake.

Runs of homozygosity and cross-generational inbreeding of Iranian fat-tailed sheep.

The Lori-Bakhtiari fat-tailed sheep is one of the most important heavyweight native breeds of Iran. The breed is robust and well-adapted to semi-arid regions and an important resource for smallholder farms. An established nucleus-based breeding scheme is used to improve their production traits but there is an indication of inbreeding depression and loss of genetic diversity due to selection. Here, we estimated the inbreeding levels and the distribution of runs of homozygosity (ROH) islands in 122 multi-generational female Lori-Bakhtiari from different half-sib families selected from a breeding station that were genotyped on the 50k array. A total of 2404 ROH islands were identified. On average, there were 19.70 ± 1.4 ROH per individual ranging between 6 and 41. The mean length of the ROH was 4.1 ± 0.14 Mb. There were 1999 short ROH of length 1-6 Mb and another 300 in the 6-12 Mb range. Additionally long ROH indicative of inbreeding were found in the ranges of 12-24 Mb (95) and 24-48 Mb (10). The average inbreeding coefficient (FROH) was 0.031 ± 0.003 with estimates varying from 0.006 to 0.083. Across generations, FROH increased from 0.019 ± 0.012 to 0.036 ± 0.007. Signatures of selection were identified on chromosomes 2, 6, and 10, encompassing 55 genes and 23 QTL associated with production traits. Inbreeding coefficients are currently within acceptable levels but across generations, inbreeding is increasing due to selection. The breeding program needs to actively monitor future inbreeding rates and ensure that the breed maintains or improves on its current levels of environmental adaptation.

Relative contribution of Imprinting, X chromosome and Litter effects to phenotypic variation in economic traits of sheep.

Data on Zandi sheep were analysed to quantify maternal and paternal imprinting, X chromosome and litter effects' contribution to phenotypic variation in birth weight (BW), weaning weight (WW), growth rate (GR), Kleiber ratio (KR), efficiency of growth (EF) and relative growth rate (RGR). To this end, a two-step approach was adopted. In the first step, each trait was analysed with a series of 16 animal models, which were identical for fixed and autosomal additive genetic effects but differed for combinations of maternal permanent environmental, maternal genetic, X chromosome and litter effects. For each trait, the best model was selected by the Akaike information criterion (AIC) and likelihood ratio tests (LRTs). In the second step, three additional models were fitted by adding maternal imprinting, paternal imprinting or both (models 17, 18 and 19) to the best model selected in the first step. Estimators of bias, dispersion and accuracy of breeding values estimated within 19 models with whole, and partial data were used to evaluate how well were the 19 models in estimating breeding values for the animals when their records were masked. For all traits studied, fitting the litter effect led to a better data fit. Also, it resulted in noticeable decreases in residual variance and other maternal variances. For growth traits, models containing the X-linked effects fitted the data substantially better than corresponding models without the X-linked effects. For BW, WW and GR, estimates of X-linked heritability ( h s 2 ) ranged between 0.09 (GR) and 0.14 (BW). Ignoring X-linked effects from the genetic evaluation model resulted in significant inflated autosomal additive genetic variance. For BW, WW, EF and RGR, models containing the imprinting effects provided a better fit of the data than otherwise identical models. Imprinting effects contributed significantly to the phenotypic variation of these traits in a range between 5% (RGR) and 8% (BW, WW). A sharp decline was observed in autosomal additive genetic variance following including imprinting effects in the model (27% to 40% depending on the trait). The least bias and dispersion, as well as greater accuracies for breeding values of focal animals, were for a model which included imprinting, X-linked and litter effects. It was concluded that imprinting, X-linked and litter effects need to be included in the genetic evaluation models for growth and efficiency-related traits of Zandi lambs.

Genome-wide association study for postweaning weight traits in Lori-Bakhtiari sheep.

Marker-assisted selection is an effective method in novel animal breeding programs. This study was conducted to perform a genome-wide association study to detect candidate genes and quantitative trait loci associated with postweaning weight traits in meat-type sheep. Body weight records were collected during 29 years (1989 to 2017) in Lori-Bakhtiari sheep flock of the Shooli Breeding Station in Iran. A total of 132 animals were selected based on estimates of breeding values (EBVs) for body weight, using two-tailed and random selection strategies. Genomic DNA was extracted from whole blood samples. The samples were genotyped using Illumina OvineSNP50 BeadChip. De-regressed EBVs for postweaning body weight traits were used as pseudo-phenotypes in a genome-wide association study. One SNP on chromosome 10 (rs406324209) and two SNPs on chromosome 13 (rs401963094 and rs418761613) were significantly (Bonferroni-adjusted p-values < 0.05) associated with postweaning body weight traits. The significant variants accounted for 0.20% and 0.48% of the total genetic variances for 6- and 9-month body weights, respectively. Genomic heritabilities estimated for 6-, 9- and 12-month weights and postweaning weight gain were 0.28 ± 0.34, 0.35 ± 0.29, 0.37 ± 0.34, and 0.16 ± 0.33, respectively. Two significant SNPs were located within the ATP8A2 and PLXDC2 genes, on chromosomes 10 and 13, respectively. Based on the known gene ontologies, both ATP8A2 and PLXDC2 could be considered as candidate genes for postweaning body weight traits.

Genetic parameters and trends for litter size in Markhoz goats / Parámetros genéticos y tendencias para el tamaño de la camada en cabras Markhoz / Parâmetros genéticos e tendências para o tamanho da ninhada em cabras Markhoz

Abstract Background: Prolificacy has a high economic value and is one of the most important reproduction traits in small ruminants such as Markhoz goats, which is an endangered breed. Objective: To estimate genetic, environmental and phenotypic trends, and genetic parameters for litter size (LS), as a prolificacy trait of Markhoz goat does. Methods: The study was conducted using kidding records of 3,064 Markhoz does from 1992 to 2015. Genetic parameters were estimated with a logit link function using ASReml software, fitting a repeatability animal model, with birth year, kidding year, and kidding age as fixed effects, and direct additive genetic and permanent environmental effects as random effects. Genetic, phenotypic and environmental trends were estimated by using regression of breeding value, phenotypic value and environmental deviation averages on birth year, respectively. Results: Low and negligible heritability (0.002), repeatability (0.054) and coefficient of permanent environment (0.052) were estimated for LS, which indicates low possibility to achieve rapid genetic progress through phenotypic selection. A non-significant -actually, zero- genetic trend (0.00004 heads/kidding/year), and significant negative phenotypic and environmental trends (-0.01097 and -0.01100 heads/kidding/year, respectively) were estimated in this study. Conclusion: Optimizing environmental conditions and application of high-throughput technologies in selection programs could help reducing the extinction risk of Markhoz breed.

Resumen Antecedentes: La prolificidad tiene un alto valor económico y es uno de los rasgos reproductivos más importantes en pequeños rumiantes, tales como la raza caprina Markhoz. que está en peligro de extinción. Objetivo: Estimar las tendencias genéticas, ambientales y fenotípicas, y los parámetros genéticos para el tamaño de la camada (LS), como rasgo de prolificidad de las hembras de cabras Markhoz. Métodos: Se utilizaron registros de partos de 3.064 hembras Markhoz, de 1992 a 2015. Los parámetros genéticos se estimaron con una función de enlace logit utilizando el software ASReml, ajustando un modelo animal de repetibilidad, con el año de nacimiento, el año del parto y la edad al parto como efectos fijos, y genéticos aditivos directos y efectos ambientales permanentes como efectos aleatorios. Las tendencias genéticas, fenotípicas y ambientales se estimaron mediante la regresión del valor de la cría, el valor fenotípico y los promedios de desviación ambiental en el año de nacimiento, respectivamente. Resultados: Se estimó la heredabilidad -baja e insignificante- (0,002), repetibilidad (0,054) y coeficiente de ambiente permanente (0,052) para LS, lo que indica una baja posibilidad de lograr un progreso genético rápido a través de selección fenotípica. Se estimó una tendencia genética no significativa -de hecho, cero- (0,00004 cabezas/parto/año) y se estimaron tendencias fenotípicas y ambientales negativas significativas (-0,01097 y -0,01100 cabezas/parto/año, respectivamente) en el período estudiado. Conclusión: La optimización de las condiciones ambientales y la aplicación de tecnologías de alto rendimiento en los programas de selección podrían reducir el riesgo de extinción de la raza Markhoz.

Resumo Antecedentes: A prolificidade tem um elevado valor económico e é uma das mais importantes características de reprodução nos pequenos ruminantes, como a raça de caprinos Markhoz, que está em perigo de extinção. Objetivo: Estimar tendências genéticas, ambientais e fenotípicas, e parâmetros genéticos para o tamanho da serapilheira (LS), como um traço de proliferação de cabras Markhoz. Métodos: Este estudo foi realizado usando registros de parto de 3.064 Markhoz entre 1992 e 2015. Os parâmetros genéticos foram estimados com uma função logit link utilizando o software ASReml, ajustando um modelo animal de repetibilidade, com o ano de nascimento, o ano de parto ea idade de caçoar como efeitos fixos e efeitos ambientais diretos e genéticos diretos como efeitos aleatórios. As tendências genéticas, fenotípicas e ambientais foram estimadas pela regressão do valor de criação, valor fenotípico e médias de desvio ambiental no ano de nascimento, respectivamente. Resultados: Herdabilidade baixa e negligenciável (0,002), repetibilidade (0,054) e coeficiente de ambiente permanente (0,052) foram estimadas para LS, o que indica baixa possibilidade de alcançar um progresso genético rápido através da seleção fenotípica. Foi estimada uma tendência genética não-significativa e realmente zero (0,00004 cabeças/parto/ano) e tendências fenotípicas e ambientais negativas significativas (-0,01097 e -0,01100 cabeças/parto/ano, respectivamente) no período estudado. Conclusão: Parece que a otimização das condições ambientais ea aplicação de tecnologias de alto rendimento em programas de seleção poderiam reduzir o risco de extinção para a raça Markhoz.

Screening for Causative Mutations of Major Prolificacy Genes in Iranian Fat-Tailed Sheep.

BACKGROUND: The presence of different missense mutations in sheep breeds have shown that the bone morphogenetic protein receptor 1B (BMPR1B), bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) genes play a vital role in ovulation rate and prolificacy in ewes. Therefore, the present study aims to investigate BMPR1B, BMP15 and GDF9 gene mutations in prolific ewes of Iranian fat-tailed Lori-Bakhtiari sheep. MATERIALS AND METHODS: In the present experimental study, genomic DNA was extracted from whole blood of 10 prolific Lori-Bakhtiari ewes with at least two twinning records in the first four parities to identify point mutations of the BMPR1B, BMP15 and GDF9 genes, using DNA sequencing. RESULTS: The results obtained from DNA sequencing showed a new synonymous mutation (g.66496G>A) in exon 8 of the BMPR1B gene, without any amino acid change. Sequencing of the BMP15 gene revealed a deletion of 3 bp (g.656_658delTTC) in exon 1, leading to an amino acid deletion (p.Leu19del). Four single nucleotide polymorphisms (G1:g.2118G>A, G2:g.3451T>C, G3:g.3457A>G and G4:g.3701G>A), were detected in exons 1 and 2 of the GDF9 gene, two of which caused amino acid substitutions (G1: p.87Arg>His and G4: p.241Glu>Lys). These amino acid alterations are proposed to have a benign impact on structure and function of the GDF9 polypeptide sequence. CONCLUSION: Three major prolificacy genes (BMPR1B, BMP15 and GDF9) were polymorphic in Lori-Bakhtiari sheep, although none of the major causative mutation was detected in this sheep type. Further studies using high throughput methods such as genome-wide association study (GWAS) and evaluation of other candidate genes are necessary in the future.

Statistical properties of proportional residual energy intake as a new measure of energetic efficiency.

Traditional ratio measures of efficiency, including feed conversion ratio (FCR), gross milk efficiency (GME), gross energy efficiency (GEE) and net energy efficiency (NEE) may have some statistical problems including high correlations with milk yield. Residual energy intake (REI) or residual feed intake (RFI) is another criterion, proposed to overcome the problems attributed to the traditional ratio criteria, but it does not account for production or intake levels. For example, the same REI value could be considerable for low producing and negligible for high producing cows. The aim of this study was to propose a new measure of efficiency to overcome the problems attributed to the previous criteria. A total of 1478 monthly records of 268 lactating Holstein cows were used for this study. In addition to FCR, GME, GEE, NEE and REI, a new criterion called proportional residual energy intake (PREI) was calculated as REI to net energy intake ratio and defined as proportion of net energy intake lost as REI. The PREI had an average of -0·02 and range of -0·36 to 0·27, meaning that the least efficient cow lost 0·27 of her net energy intake as REI, while the most efficient animal saved 0·36 of her net energy intake as less REI. Traditional ratio criteria (FCR, GME, GEE and NEE) had high correlations with milk and fat corrected milk yields (absolute values from 0·469 to 0·816), while the REI and PREI had low correlations (0·000 to 0·069) with milk production. The results showed that the traditional ratio criteria (FCR, GME, GEE and NEE) are highly influenced by production traits, while the REI and PREI are independent of production level. Moreover, the PREI adjusts the REI magnitude for intake level. It seems that the PREI could be considered as a worthwhile measure of efficiency for future studies.

A new mutation in exon 2 of the bone morphogenetic protein 15 gene is associated with increase in prolificacy of Mehraban and Lori sheep.

Some mutations in bone morphogenetic protein 15 (BMP15) gene have been known to be associated with prolificacy in various breeds of sheep. Polymorphism of BMP15 gene exon 2 was studied in 100 Mehraban and 100 Lori ewes, using PCR-SSCP and DNA sequencing methods. A new point mutation (G → A) was found at position 57 of the amplified 312-b fragment of BMP15 gene exon 2. The frequencies of the AG and GG genotypes were 69.4 and 30.6 % in Mehraban and 44.7 and 55.3 % in Lori ewes, respectively. No individual carrying the AA genotype was found in the studied population. The allelic frequencies for A and G alleles were 34.7 and 65.3 % in Mehraban and 22.4 and 77.6 % in Lori ewes, respectively. Average litter size in the AG genotype (1.56) was significantly (P < 0.01) higher than the GG ewes (1.08). The results of the present study indicated the potential of the observed SNP in exon 2 of BMP15 for further exploitation in marker-assisted selection to improve reproduction efficiency of Mehraban and Lori sheep.

Effect of Zinc Source on Hematological, Metabolic Parameters and Mineral Balance in Lambs.

This experiment was conducted to study the effects of different sources of zinc (Zn) on blood metabolites and balances of some minerals in lambs. In the first part, 20 6-7-month-old lambs were randomly allotted to four treatments including (1) basal diet containing 22.47 mg Zn/kg DM without supplementary Zn (control), (2) basal diet + 40 mg Zn/kg DM as ZnSO4 (ZnSO4 40), (3) basal diet + 20 mg Zn/kg DM as Zn-proteinate (Zn-Pro 20), and (4) basal diet + 40 mg Zn/kg DM as Zn-proteinate (Zn-Pro 40). Blood samples were taken on days 0, 28, and 65 before morning feeding. In the second part, four lambs from each treatment were randomly transferred to metabolic cages to evaluate the effects of different sources of Zn on N, Zn, Fe, and Cu retentions. This trial consisted of 18 days, with the first 12 days as the adaptation period followed by 6 days of sample collection. The results of this study showed that the source of Zinc had no significant effect on the analyzed parameters. Average daily gain and feed efficiency were improved by Zn supplementation (P < 0.05). Daily feed intake, plasma glucose, Fe and Cu concentrations, serum total antioxidant capacity, red blood cell count, packed cell volume, and hemoglobin concentration did not differ significantly between treatments (P > 0.05). Plasma Zn concentration, alkaline phosphatase (ALP) and bone-specific alkaline phosphatase (BALP) activity, and white blood cell and lymphocyte count differed significantly between control and Zn-supplemented groups (P < 0.05) as Zn supplementation improved these parameters. Nitrogen, Fe, and Cu retentions did not differ between treatments (P > 0.05). Zinc retention showed a significant difference between control and Zn-supplemented groups (P < 0.05), but there were no significant differences among the Zn-supplemented groups. The results of this study show that Zn supplementation improved performance and zinc retention in lambs. However, there were no significant differences between zinc sources used in this study.