Turkish Cattle Breeds Research Articles

Analysing the genetic diversity and population structure of five native Turkish cattle breeds using SNP data.

The conservation and sustainable utilization of cattle genetic resources necessitate a comprehensive understanding of their genetic diversity and population structure. This study provides an analysis of five native Turkish cattle breeds: Anatolian Black (ANB), Turkish Grey (TUR), Anatolian Southern Yellow (ASY), East Anatolian Red (EAR), and South Anatolian Red (SAN) using 50 K SNP data. These breeds were compared with three European breeds, Simmental (SIM), Holstein (HOL), and Jersey (JER), and three Asian Zebu breeds: Arabic Zebu (ZAR), Nelore (NEL), and Red Sindhi (RSI). Genetic diversity indices demonstrated moderate heterogeneity among the breeds, with TUR exhibiting the highest observed heterozygosity (Ho = 0.35). Wright's Fst values indicated significant genetic differentiation, particularly between Turkish breeds and both European (Fst = 0.035-0.071) and Asian breeds (Fst = 0.025-0.150). Principal component analysis distinguished the unique genetic profiles of each breed cluster. Admixture analysis revealed degrees of shared genetic ancestry, suggesting historical gene flow between Turkish, European, and Asian breeds. Analysis of molecular variance (AMOVA) attributed approximately 58% of the variation to population differences. Nei's genetic distances highlighted the closer genetic relatedness within Turkish breeds (distance ranges between 0.032 and 0.046) and suggested a more relative affinity of TUR with European breeds. The study's phylogenetic assessments elucidate the nuanced genetic relationships among these breeds, with runs of homozygosity (ROH) analysis indicating patterns of ancestral relatedness and moderate levels of inbreeding, particularly evident in Turkish breeds. Our findings provide valuable insights into the genetic landscape of Turkish cattle, offering a crucial foundation for informed conservation and breeding strategies aimed at preserving these breeds' genetic integrity and heritage.

Mitochondrial DNA diversity of D-loop region in three native Turkish cattle breeds.

This study aimed to reveal the genetic variability of mitochondrial DNA (mtDNA) displacement-loop (D-loop) region in 62 animals belonging to three native Turkish cattle breeds, namely Anatolian Black (AB), East Anatolian Red (EAR) and Zavot (ZAV), and to conduct phylogenetic relationship analyses to obtain deeper information on their genetic origin and breeding history by comparison of 6 taurine and 11 indicine breeds, together with 66 polymorphic sites, a total of 31 haplotypes, of which 15, 10 and 6 were detected in AB, EAR and ZAV, respectively. Mean nucleotide and haplotype diversity were 0.01 and 0.891, respectively, whereas the genetic differentiation derived from Wright's index was 0.174 across the breeds. A significant level of total variation (17.42 %) was observed among breeds in molecular variance analysis. Six main haplogroups (T, T1, T2, T3, Q and I2) were detected in Anatolian cattle populations, where T3 was the most frequent among breeds (43.55 %), whereas I2, an indicine specific haplogroup, was observed only in ZAV. At the breed level, phylogenetic analyses supported by 198 sequences of 17 cattle breeds and 3 outgroup species retrieved from the GenBank clustered native Turkish cattle breeds with the taurine group rather than the indicine one, as expected. However, indicine admixture at low frequency (8.89 %) was detected in the ZAV breed for the first time due to more likely gene flow from indicine cattle breeds raised in neighbour countries, particularly Iran. This finding should be further investigated in all native Turkish and indicine cattle breeds from nearby countries to clarify gene flow and indicine admixture in Anatolian cattle.

Future Perspective of NGS Data for Evaluation of Population Genetic Structure in Turkish Cattle

Developments in sequencing and SNP chip technologies have enabled scientists to obtain high-density genomic data from different livestock species, including cattle. Moreover, many bioinformatics tools are available to analyse high-density genomic data. Via these tools, several statistical approaches such as Principal Component Analysis and clusterin-based analyses could be conducted to reveal the genetic structure of cattle populations. However, revealing the genetic structure and selection signatures of Turkish cattle breeds is a new area of research, since the previous studies are limited with a few microsatellite data. On the other hand, rearing in different geographical and environmental conditions for a long period could possibly lead to more genetic variation in native Turkish cattle breeds compared to high-yielding culture breeds. These variations obviously cannot be detected by limited number of microsatellite markers, while Next Generation Sequencing is promising for further population structure studies. Hence this review aims to summarise previous studies and give a perspective of Next Generation Sequencing possibilities to reveal the population structure of Turkish cattle for further studies.

Genotypic structure of four cattle breeds raised in Turkey by loci related to several diseases

This study aims to reveal the genotypic structure of four cattle breeds; Holstein (HS), Turkish Grey Steppe (TGS), Anatolian Black (AB) and East Anatolian Red (EAR), raised in Turkey in terms of CD14, MBL, ITGB6, SLC11A1 and TLR2 genes and to evaluate their usefulness in Marker Assisted Selection (MAS). It also assesses whether the loci associated with resistance to diseases are suitable for phylogenetic analysis. Desired alleles and/or genotypes were detected in native Turkish cattle breeds at different frequencies in terms of polymorphisms of CD14, MBL, ITGB6, SLC11A1 and TLR2 genes which were previously reported to be associated with mastitis, foot-and-mouth disease and tuberculosis. These variations offer opportunities to improve selection strategies against diseases in the future. These results preliminary indicate that associated studies between these variations and disease resistance in native Turkish cattle breeds should be conducted. On the other hand, phylogenetic tree constructed based on genetic distance clearly separated native Turkish cattle breeds from HS breed. The gene regions related to diseases can be used to distinguish native cattle breeds from exotic ones.

A comprehensive review on genetic diversity and phylogenetic relationships among native Turkish cattle breeds based on microsatellite markers

A comprehensive review on genetic diversity and phylogenetic relationships among native Turkish cattle breeds based on microsatellite markers

Phylogenetic relationships of native Turkish cattle breeds using microsatellite markers

A total of 20 microsatellite DNA markers were used for genetic characterization and determination of phylogenetic relationships of native cattle breeds of Turkey, including the Anatolian Grey (AG), Anatolian Black (AB), South Anatolian Red (SAR), East Anatolian Red (EAR), Southern Anatolian Yellow (SAY), and Zavot (ZAV). DNA samples were isolated from 271 blood samples using an organic method. Amplified polymerase chain reaction products were separated by capillary electrophoresis and genotypes were determined for 20 microsatellites. A total of 269 different alleles were determined. The lowest (7.80) and highest (10.80) mean allele numbers were observed for the ZAV and SAY populations, respectively. TGLA122 was the most polymorphic locus; however, only 7 different alleles were observed for INRA005. A total of 40 different private alleles were determined. The general FIS values were between 0.034 and 0.123. Due to the close location to the domestication center, higher genetic diversities were observed. The observed genetic diversities and the results of the phylogenetic analyses were in agreement with evolutionary history and the geographical origins of Turkish native cattle breeds.

Population structure and ancestry of Qinchuan cattle.

The aim of this study was to estimate population structure and ancestry of Qinchuan cattle by genotyping 27 individuals using the GeneSeek HD 77k BeadChip, and another 1355 cattle representing breeds distributed worldwide, which had been genotyped using the Illumina Bovine 50k BeadChip. Qinchuan cattle were characterized by a dominant Bos taurus ancestry, accompanied by a considerable proportion of Bos indicus ancestry based on principal components analysis and supervised admixture analysis. A small proportion of Bos javanicus ancestry was detected as well. A similar admixture pattern in both Qinchuan and Turkish cattle breeds reflects their similar degrees of zebu introgression. Our study presents a relatively clear view of the population structure and ancestry of Qinchuan cattle, serving to benefit our understanding of this breed and leading to better targeted conservation approaches moving forward.

Determination of Toll-Like Receptor 1 Gene Polymorphisms in Zavot, Turkish Grey, East Anatolian Red, Anatolian Black and South Anatolian Red Cattle Breeds

Toll-like receptors (TLRs) play an important role in non-specific immunity against different infectious agents such as bacterium or parasite. The aim of this work was to investigate the allele and genotype frequencies of three single-nucleotide polymorphisms (SNPs) in bovine TLR1 gene in native Turkish cattle breeds. DNA samples were extracted using the phenol chloroform protocol from 77 Zavot, 60 Turkish Grey, 51 East Anatolian Red, 69 Anatolian Black and 46 South Anatolian Red cattle. Target regions of the TLR1 gene were digested BsII and HpyI88III restriction enzymes. Results showed that the (A) allele frequency had higher in all native Turkish cattle breeds of the TLR1-G1409A locus. The (F) allele frequency was found to be higher compared to (E) allele in the TLR1-G1550A site. The frequencies of both (C) and (T) alleles were close to each other in the TLR1-C632T site. In conclusion genetic polymorphisms exist in Turkish native cattle populations in terms of known TLR1 variants.

Growth hormone (GH), prolactin (PRL), and diacylglycerol acyltransferase (DGAT1) gene polymorphisms in Turkish native cattle breeds

Abstract: The aim of this study was to determine the genetic diversity of 4 native Turkish cattle breeds, based on the growth hormone (GH), prolactin (PRL), and diacylglycerol acyltransferase (DGAT1) genes. In order to study the polymorphisms in these genes, the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method was performed. A 329-bp fragment and a 223-bp fragment of GH, a 156-bp fragment of PRL, and a 411-bp fragment of DGAT1, thus 4 loci of 3 genes, were amplified via PCR. These fragments were then restricted with the enzymes MspI, Alu1, RsaI, and CfrI, respectively. In this study, 2 types of alleles, ( + ) and ( - ) for the GH-MspI, L and V for the GH-Alu1, A and B for the PRL, and K and A for the DGAT1 loci, were observed. The results of the present study will contribute to the polymorphism data on the world's cattle breeds. Furthermore, the above-mentioned allele frequencies of Turkish native breeds are evaluated in relation to their genetic relatedness, and to infer their milk production properties on the basis of the available literature. In turn, these results can be utilized for future breeding programs of dairy cattle in Turkey.

Y chromosome analysis of native Turkish cattle breeds by microsatellite markers

The aim of this study was to determine phylogenetic relationships of 6 native cattle breeds of Turkey using 7 Y chromosome-specific microsatellite DNA markers. DNA samples were isolated from Anatolian Black, Anatolian Grey, South Anatolian Red, Native Southern Anatolian Yellow, East Anatolian Red, and Zavot cattle using a standard phenol/chloroform method. PCR products were separated by capillary electrophoresis and marker genotypes were determined. A total of 41 different alleles were observed. The mean allele number was 5.86 and mean FIS value was 0.427 for all populations. The INRA189 locus was monomorphic for all populations except for South Anatolian Red. UMN0307 and INRA124 loci were monomorphic in the East Anatolian Red and Anatolian Black populations, respectively. The Anatolian Black population was assigned to its own population at a maximum level (78.26%). In this study, a taurine-specific allele was identified in the INRA124 locus. Zebu- and taurine-specific alleles were observed in BM861, INRA189, and UMN0103 loci. Allele 134 in INRA124 seems to be a specific allele of taurine or zebu. The alleles of the UMN0307 and UMN0504 loci need to be investigated for zebu- or taurine-specific alleles. The resulting neighbor-joining tree and structure analysis suggested that the breeds analyzed are consistent with their modern geographical locations.

Genetic Characterization of Turkish Cattle Breeds by Microsatellite Markers: Usefulness for Parentage Testing

Objective of this study was to evaluate microsatellite markers in paternity testing of native cattle breeds in Turkey. Blood samples were collected from Anatolian Black (n=51), Anatolian Grey (n=54), South Anatolian Red (n=51), Native Southern Anatolian Yellow (n=51), East Anatolian Red (n=45) and Zavot (n = 19) cattle. From the blood samples DNA was isolated by using a standard phenol/ chloroform method. A total of 20 microsatellite loci were selected from a FAO/ISAG-suggested list. Polymerase chain reaction products were separated by capillary electrophoresis and marker genotypes were determined by fragment analysis. In statistical analyses, allel numbers, observed (Ho) and expected (He) heterozygosities, deviation from Hardy-Weinberg Equilibrium and probability of exclusion (PE) at each microsatellite locus were calculated. A total of 269 different alleles were observed and the mean allele was identified as 13.45. Mean Ho and He values were observed as 0.619-0.852 and 0.669-0.877, respectively. The results indicated that the microsatellite test panel including the most informative 7 loci had total PE value of >0.9999 in each populations and can thereby be used for parentage testing studies of native cattle breeds in Turkey.

Genetic diversity of native Turkish cattle breeds: Mantel, AMOVA and bottleneck analysis

Objective of this study was to evaluate potential extinction risk of Turkish native cattle breeds using Mantel and AMOVA tests and Bottleneck analysis. A total of 271 DNA samples were isolated from Anatolian Black, Anatolian Grey, South Anatolian Red, Native Southern Anatolian Yellow, East Anatolian Red and Zavot cattle. Genotypes of twenty microsatellites were determined by capillary electrophoresis and fragment analysis. In this study, a total of 269 different alleles were detected. The maximum and minimum numbers of total alleles were observed in TGLA122 (26 allele) and INRA005 (7 allele) loci, respectively. The highest average observed and expected heterozygosity values were determined as 0.619–0.852 and 0.669–0.877, respectively. The average FIS value was 0.068. Results of AMOVA and Mantel tests illustrated statistically significant differences in populations (p

Beta-casein A1/A2, kappa-casein and beta-lactoglobulin polymorphisms in Turkish cattle breeds

Abstract. In this study, the genetic diversity of three milk protein genes namely beta-casein, kappacasein and beta-lactoglobulin was estimated in Turkish cattle breeds. Based on these genes, breeds in Turkey have been grouped as: 1) Eastern Anatolian Red, Anatolian Black and Southern Anatolian Red and 2) Turkish Grey, Turkish Holstein and Holstein Candidate Bulls. B alleles of the three studied genes, which were reported to be positively related with cheese yield and quality, seemed to be low-intermediate for beta-casein and kappa-casein but relatively high for beta-lactoglobulin in the first group of Turkish breeds compared to other breeds of the world. The kappa-casein E allele, which has a negative effect on cheese quality, is absent in Turkish cattle breeds, except in Holstein Candidate Bulls. Therefore, the results suggest that milk of Turkish native breeds is suitable for cheese making. Based on observations of the Turkish breeds, some suggestions were made regarding breeding practices in Turkey.

Leptin Gene Polymorphisms in Native Turkish Cattle Breeds

Summary The aim of the study was to determine leptin gene polymorphisms in South Anatolian Red (SAR), East Anatolian Red (EAR) and Turkish Grey Cattle. In the study unrelated 40 SAR, 40 EAR and 40 Turkish Grey cattle were used. Target sites in leptin gene exon 2, exon 3 and intron 2 were amplified by polymerase chain reaction (PCR). The single nucleotide polymorphism (SNP) consisting site in exon 2, exon 3 and intron 2 were determined as a result of digestion with Kpn2I, HphI and Sau3AI restriction enzymes, respectively. The highest T allele frequency related with production traits for Kpn2I polymorphism was found for SAR cattle. For HphI polymorphism, T allele frequencies were detected clearly predominant. Within each breed for Sau3AI polymorphism B and C allele frequencies that effect production traits were found to be dramatically lower than A allele frequency. As a result we can suggest that there was no clearly difference that can create any advantage in terms of leptin gene SNPs among the three native Turkish cattle breeds.

265 SEXING OF BOVINE EMBRYOS BASED ON PCR TECHNIQUE

Turkish cattle breeds are well adapted to harsh environmental and poor feeding conditions. However, their productivity is low. Increasing fertility rate and obtaining a high number of progeny from high-quality animals are important parameters in animal husbandry. The objective of the present project is to produce sexed embryos and cryopreserve them for subsequent transfer. The birth of the calves produced from the transferred embryos with sex determined prior to transfer by PCR are additional objectives of the study. To develop and optimize the PCR method, DNA was first isolated by using standard phenol-chloroform extraction from blood samples of cows and bulls to use as positive control. Then two multiplex PCR methods were developed using one autosomal (bovine 1.715 satellite locus which produces 216 bp long PCR product), and two Y-chromosome specific loci BRY4.a (300 bp) and BRY1 (300 bp). Both multiplexes include bovine 1.715 satellite locus, and they either include BRY4.a or BRY.1 as the second locus. Female individuals produce one PCR band, whereas male individuals produce two PCR bands. Bovine parthenogenetic blastocysts were used to test these two multiplex PCR methods. Immature bovine oocytes were aspirated from slaughterhouse material and in vitro matured in tissue culture medium-199 (TCM-199) supplemented with 10% FCS, sodium pyruvate, EGF, bLH, bFSH and penicillin/streptomycin for 18 h at 39°C and 5% CO2 in humidified air. After removing the cumulus cells of matured oocytes (MII), meiotic spindles and first polar bodies were removed. Oocyte-cell complexes were fused by one 30 μs pulse of 133V/500 μm. All fusion units were subjected to chemical activation. Afterwards, parthenogenetic oocytes were cultured in Sage cleavage® medium supplemented with 8 mg mL–1 BSA for 72 h and then developing embryos were cultured in Sage blastocyst® media supplemented with 4 mg mL–1 BSA + 5% FCS for 4 additional days. Then they were stored at –20°C until DNA extraction. For DNA extraction two methods (Park et al. 2001 Theriogenology 55, 1843–1853; Tshimangadzo et al. 2004 Biol. Reprod. 71, 1671–1676) were employed to test their efficiency in our laboratory conditions, and we got better results with the former method. Repeated PCR tests of these parthenogenetic blastocysts were carried out and tests revealed only one PCR product of 216 bp corresponding to the 1.715 satellite locus as expected. The multiplex PCR methods will also be employed for Nuclear Transfer and IVF embryos. This project is supported by Turkish Scientific and Technical Research Council-TOVAG (project no: KAMAG 107G027).

Intragenic haplotypes at the bovine <i>CSN1S1</i> locus

Abstract. A new alternative genotyping method based on PCR-SSCP was developed for direct differentiation of the CSN1S1 alleles B and C in the coding region. In addition a PCR-RFLP test based on a MaeIII restriction site in the promoter region of CSN1S1, reported in the literature as an alternative test for the differentiation of CSN1S1*B and C was used and the alleles named b and c. Genotyping of 649 animals belonging to 17 European and Turkish cattle breeds showed differences in occurrence and frequency of the alleles. CSN1S1*B occurred in all breeds with frequencies varying from 0.50 in Anatolian Blackup to 1.0 in e.g. Ayrshire. CSN1S1*b on the other hand varied from 0.63 in Jersey, 0.97 in Ayrshire to 1.0 in e.g. Angler. Comparison of the results from both typing methods and positions in the gene showed that both mutations do not always occur together. From the resulting four intragenic haplotypes (B-b, B-c, C-c and C-b) B-b is predominant in all breeds with frequencies varying from 0.3450 in Anatolian Black to 1.0 in Angler and Scottish Highland. The number of haplotypes varied from only one in Angler and Scottish Highland, two in Ayrshire, three in Asturian Valley and Turkish Grey Steppe to all four in the other 12 breeds. Correlation between allele frequencies and the geographic origin of the breeds was significant for the MaeIII promoter polymorphism.