Gene-Set Enrichment Analysis for Identifying Genes and Biological Activities Associated with Growth Traits in Dromedaries

Simple Summary This project aimed to find biological themes affecting growth in dromedaries. Candidate SNPs associated with growth were mapped to 22 genes, and 25 significant themes were identified related to growth. The main biological functions included calcium ion binding, protein binding, DNA-binding transcription factor activity, protein kinase activity, tropomyosin binding, myosin complex, actin-binding, ATP binding, receptor signaling pathway via JAK-STAT, and cytokine activity. EFCAB5, MTIF2, MYO3A, TBX15, IFNL3, PREX1, and TMOD3 genes are candidates for improving growth in camel breeding programs. Growth is an important heritable economic trait for dromedaries and necessary for planning a successful breeding program. Until now, genome-wide association studies (GWAS) and QTL-mapping have identified significant single nucleotide polymorphisms (SNPs) associated with growth in domestic animals, but in dromedaries, the number of studies is very low. This project aimed to find biological themes affecting growth in dromedaries. In the first step, 99 candidate SNPs were chosen from a previously established set of SNPs associated with body weight, gain, and birth weight in Iranian dromedaries. Next, 0.5 kb upstream and downstream of each candidate SNP were selected from NCBI (assembly accession: GCA_000803125.3). The annotation of fragments with candidate SNPs regarding the reference genome was retrieved using the Blast2GO tool. Candidate SNPs associated with growth were mapped to 22 genes, and 25 significant biological themes were identified to be related to growth in dromedaries. The main biological functions included calcium ion binding, protein binding, DNA-binding transcription factor activity, protein kinase activity, tropomyosin binding, myosin complex, actin-binding, ATP binding, receptor signaling pathway via JAK-STAT, and cytokine activity. EFCAB5, MTIF2, MYO3A, TBX15, IFNL3, PREX1, and TMOD3 genes are candidates for improving growth in camel breeding programs

Genomic prediction for growth using a low-density SNP panel in dromedary camels

For thousands of years, camels have produced meat, milk, and fiber in harsh desert conditions. For a sustainable development to provide protein resources from desert areas, it is necessary to pay attention to genetic improvement in camel breeding. By using genotyping-by-sequencing (GBS) method we produced over 14,500 genome wide markers to conduct a genome- wide association study (GWAS) for investigating the birth weight, daily gain, and body weight of 96 dromedaries in the Iranian central desert. A total of 99 SNPs were associated with birth weight, daily gain, and body weight (p-value \u3c 0.002). Genomic breeding values (GEBVs) were estimated with the BGLR package using (i) all 14,522 SNPs and (ii) the 99 SNPs by GWAS. Twenty-eight SNPs were associated with birth weight, daily gain, and body weight (p-value \u3c 0.001). Annotation of the genomic region (s) within ± 100 kb of the associated SNPs facilitated prediction of 36 candidate genes. The accuracy of GEBVs was more than 0.65 based on all 14,522 SNPs, but the regression coefficients for birth weight, daily gain, and body weight were 0.39, 0.20, and 0.23, respectively. Because of low sample size, the GEBVs were predicted using the associated SNPs from GWAS. The accuracy of GEBVs based on the 99 associated SNPs was 0.62, 0.82, and 0.57 for birth weight, daily gain, and body weight. This report is the first GWAS using GBS on dromedary camels and identifies markers associated with growth traits that could help to plan breeding program to genetic improvement. Further researches using larger sample size and collaboration of the camel farmers and more profound understanding will permit verification of the associated SNPs identified in this project. The preliminary results of study show that genomic selection could be the appropriate way to genetic improvement of body weight in dromedary camels, which is challenging due to a long generation interval, seasonal reproduction, and lack of records and pedigrees

Genomic prediction for growth using a low-density SNP panel in dromedary camels

For thousands of years, camels have produced meat, milk, and fiber in harsh desert conditions. For a sustainable development to provide protein resources from desert areas, it is necessary to pay attention to genetic improvement in camel breeding. By using genotyping-by-sequencing (GBS) method we produced over 14,500 genome wide markers to conduct a genome- wide association study (GWAS) for investigating the birth weight, daily gain, and body weight of 96 dromedaries in the Iranian central desert. A total of 99 SNPs were associated with birth weight, daily gain, and body weight (p-value \u3c 0.002). Genomic breeding values (GEBVs) were estimated with the BGLR package using (i) all 14,522 SNPs and (ii) the 99 SNPs by GWAS. Twenty-eight SNPs were associated with birth weight, daily gain, and body weight (p-value \u3c 0.001). Annotation of the genomic region (s) within ± 100 kb of the associated SNPs facilitated prediction of 36 candidate genes. The accuracy of GEBVs was more than 0.65 based on all 14,522 SNPs, but the regression coefficients for birth weight, daily gain, and body weight were 0.39, 0.20, and 0.23, respectively. Because of low sample size, the GEBVs were predicted using the associated SNPs from GWAS. The accuracy of GEBVs based on the 99 associated SNPs was 0.62, 0.82, and 0.57 for birth weight, daily gain, and body weight. This report is the first GWAS using GBS on dromedary camels and identifies markers associated with growth traits that could help to plan breeding program to genetic improvement. Further researches using larger sample size and collaboration of the camel farmers and more profound understanding will permit verification of the associated SNPs identified in this project. The preliminary results of study show that genomic selection could be the appropriate way to genetic improvement of body weight in dromedary camels, which is challenging due to a long generation interval, seasonal reproduction, and lack of records and pedigrees

Molecular Detection of Bovine Leukocytic Anaplasma Species in Isfahan, Iran

Background: A. bovis and A. phagocytophilium are leukocytotropic agents of bovine anaplasmosis. They are obligate intracellular bacteria that can infect and cause Anaplasmosis in human and animals. Therefore, this study was carried out to detect A. bovis and A. phagocytophilum in naturally infected dairy cattle in Isfahan using molecular techniques. Materials and Methods: In this study a total of 209 blood samples were collected from cattle in central part of Iran (Isfahan). The presence of A. bovis and A. phagocytophilium were examined by species-specific nested polymerase chain reaction (nPCR) based on 16S rRNA gene. Results: Out of the 209 cattle examined, 4 (1.99%) and 2 (1%) were found positive for A. bovis and A. phagocytophilium by nPCR, respectively. Conclusion: These data showed a relatively low prevalence of leukocytic Anaplasma infection in cattle in central part of Iran

Study of polymorphism in Mazandaran and Esfahan native chicken population using microsatellite markers

In order to studying of genetic variation in Mazandaran and Esfahan native chickens, twenty microsatellite markers were evaluated. These microsatellite markers were MCW0014, MCW0081, MCW0183, MCW0067, MCW0104, MCW0123, MCW0330, MCW0165, MCW0069, MCW0020, MCW0222, LEI0094, MCW0295, MCW0034, MCW0216, ADL0268, ADL0112, ADL0278 and lEI0166. Blood samples of 90 and 150 native chickens of Mazandaran and Esfahan were randomly taken respectively. Genomic DNAs were isolated through optimized and modified salting-out procedure. The number of alleles varied from 1 to 6. In Mazandaran population one locus (MCW0216) and in Esfahan population three loci (MCW0216, MCW67 and MCW222) were monomorphic. The other loci were showed appropriate polymorphism. All the loci except MCW222 and MCW165 in Mazandaran population showed deviations from Hardy-Weinberg equilibrium (

A New Approach in the Evaluation of Dairy Camels: Using Test Day Milk and Morphometric Records

In the case of camels, there is little data in the literature on the relationship between body building features and the evaluation of milk yield. In the last decade, a rising interest in camel milk has been observed due to its nutritional and health-promoting properties, resulting in a growing market demand. Despite the remarkable importance of camels, very little improvement in camel breeding and selection for dairy purposes has been achieved. The current study aimed to provide a practical approach to the evaluation of dairy dromedaries based on test day milk and morphometric records. A total of 62 Sindi dairy camels were evaluated and 4176 daily milk records were registered in February, March, April, and May 2021. She-camels were milked twice per day (at morning and evening) by hand before calf sucking. The farming system was intensive with two times feeding. Three measurements had the highest scores in assessing: udder, teats distance and placement, and teats size, which included 45 out of 100 scores. Test day milk records were analysed using a simple repeatability model with two random effects. The range of daily milk yields was estimated between 0.1 to 8.70 kg. The mean of body scores was 77.19 (CI = 74.19–80.19). Daily milk yields moderately correlated with body score (r = +0.27). Additionally, udder circumference and abdomen girth were correlated to milk production. Using test day milk records in breeding programs can be appropriate for the selection and replacement of she-camels, but due to difficulties in accessing these data, using morphometric data is a good criterion for the evaluation of dromedaries in extensive systems

A New Approach in the Evaluation of Dairy Camels: Using Test Day Milk and Morphometric Records

In the case of camels, there is little data in the literature on the relationship between body building features and the evaluation of milk yield. In the last decade, a rising interest in camel milk has been observed due to its nutritional and health-promoting properties, resulting in a growing market demand. Despite the remarkable importance of camels, very little improvement in camel breeding and selection for dairy purposes has been achieved. The current study aimed to provide a practical approach to the evaluation of dairy dromedaries based on test day milk and morphometric records. A total of 62 Sindi dairy camels were evaluated and 4176 daily milk records were registered in February, March, April, and May 2021. She-camels were milked twice per day (at morning and evening) by hand before calf sucking. The farming system was intensive with two times feeding. Three measurements had the highest scores in assessing: udder, teats distance and placement, and teats size, which included 45 out of 100 scores. Test day milk records were analysed using a simple repeatability model with two random effects. The range of daily milk yields was estimated between 0.1 to 8.70 kg. The mean of body scores was 77.19 (CI = 74.19–80.19). Daily milk yields moderately correlated with body score (r = +0.27). Additionally, udder circumference and abdomen girth were correlated to milk production. Using test day milk records in breeding programs can be appropriate for the selection and replacement of she-camels, but due to difficulties in accessing these data, using morphometric data is a good criterion for the evaluation of dromedaries in extensive systems

Genome-Wide Diversity, Population Structure and Demographic History of Dromedaries in the Central Desert of Iran

The development of camel husbandry for good production in a desert climate is very important, thus we need to understand the genetic basis of camels and give attention to genomic analysis. We assessed genome-wide diversity, linkage disequilibrium (LD), effective population size (Ne) and relatedness in 96 dromedaries originating from five different regions of the central desert of Iran using genotyping-by-sequencing (GBS). A total of 14,522 Single Nucleotide Polymorphisms (SNPs) with an average minor allele frequency (MAF) of 0.19 passed quality control and filtering steps. The average observed heterozygosity in the population was estimated at 0.25 ± 0.03. The mean of LD at distances shorter than 40 kb was low ( = 0.089 ± 0.234). The camels sampled from the central desert of Iran exhibited higher relatedness than Sudanese and lower than Arabian Peninsula dromedaries. Recent Ne of Iran's camels was estimated to be 89. Predicted Tajima's D (1.28) suggested a bottleneck or balancing selection in dromedary camels in the central desert of Iran. A general decrease in effective and census population size poses a threat for Iran's dromedaries. This report is the first SNP calling report on nearly the chromosome level and a first step towards understanding genomic diversity, population structure and demography in Iranian dromedaries

Genome-Wide Diversity, Population Structure and Demographic History of Dromedaries in the Central Desert of Iran

The development of camel husbandry for good production in a desert climate is very important, thus we need to understand the genetic basis of camels and give attention to genomic analysis. We assessed genome-wide diversity, linkage disequilibrium (LD), effective population size (Ne) and relatedness in 96 dromedaries originating from five different regions of the central desert of Iran using genotyping-by-sequencing (GBS). A total of 14,522 Single Nucleotide Polymorphisms (SNPs) with an average minor allele frequency (MAF) of 0.19 passed quality control and filtering steps. The average observed heterozygosity in the population was estimated at 0.25 ± 0.03. The mean of LD at distances shorter than 40 kb was low (r2 = 0.089 ± 0.234). The camels sampled from the central desert of Iran exhibited higher relatedness than Sudanese and lower than Arabian Peninsula dromedaries. Recent Ne of Iran’s camels was estimated to be 89. Predicted Tajima’s D (1.28) suggested a bottleneck or balancing selection in dromedary camels in the central desert of Iran. A general decrease in effective and census population size poses a threat for Iran’s dromedaries. This report is the first SNP calling report on nearly the chromosome level and a first step towards understanding genomic diversity, population structure and demography in Iranian dromedaries

Identification of Candidate Genes for Pigmentation in Camels Using Genotyping-by-Sequencing

Simple Summary The coat color of dromedary is usually uniform and varies from black to white. We identified 9 significant SNPs associated with white color, and the 13 significant SNPs associated with black color using genotyping-by-sequencing (GBS). Among candidate genes, SNAI1 that interacts with MCIR, ASIP and KIT genes plays a key role in the melanin biosynthetic and pigmentation biological process and melanogenesis biological pathway. The coat color of dromedary is usually uniform and varies from black to white, although dark- to light-brown colors are the most common phenotypes. This project was designed to gain knowledge on novel color-related variants using genotyping-by-sequencing (GBS). The association between the SNPs and coat color was tested using MLM (mixed linear models) with kinship matrix. Three GWAS models including white color vs. non-white color, black vs. non-black color, and light-brown vs. dark-brown color were performed. There were no distinct genetic clusters detected based on the color phenotypes. However, admixture occurred among all individuals of the four different coat color groups. We identified nine significant SNPs associated with white color after Bonferroni correction, located close to ANKRD26, GNB1, TSPYL4, TEKT5, DEXI, CIITA, TVP23B, CLEC16A, TMPRSS13, FXYD6, MPZL3, ANKRD26, HFM1, CDC7, TGFBR3, and HACE1 genes in neighboring flanking regions. The 13 significant SNPs associated with black color and the candidate genes were: CAPN7, CHRM4, CIITA, CLEC16A, COL4A4, COL6A6, CREB3L1, DEXI, DGKZ, DGKZ, EAF1, HDLBP, INPP5F, MCMBP, MDK, SEC23IP, SNAI1, TBX15, TEKT5, TMEM189, trpS, TSPYL4, TVP23B, and UBE2V1. The SNAI1 gene interacted with MCIR, ASIP and KIT genes. These genes play a key role in the melanin biosynthetic and pigmentation biological process and melanogenesis biological pathway. Further research using a larger sample size and pedigree data will allow confirmation of associated SNPs and the identified candidate genes