Polymorphisms in toll-like receptor (TLR) 1, 4, 9 and SLC11A1 genes and their association with paratuberculosis susceptibility in Holstein and indigenous crossbred cattle in Turkey.

Mycobacterium avium subsp. paratuberculosis (MAP) causes major problem in a wide range of animal species. In ruminant livestock including cattle, it causes a chronic disease called Johne's disease, or paratuberculosis (pTB) which is currently considered as potential zoonosis, causing Crohn's disease in humans. MAP infection susceptibility is suspected to be controlled by host genetics. Thus, selecting individuals according to their genetic structure could help to obtain bovine populations that are increasingly resistant to MAP infection. The aim of the present work was to investigate the association between toll-like receptor (TLR) 1 (+1380 G/A), TLR1 (+1446 C/A), TLR4 (+10 C/T), TLR9 (+1310 G/A) and solute carrier family 11 member 1 (SLC11A1) (+1066 C/G) mutations and MAP infection status in 813 cattle comprising East Anatolian Red crossbred, Anatolian Black crossbred and Holstein breed. TLR1 (+1380 G/A) mutation showed an association with bovineMAP (P<0.05). For the TLR1 (+1380 G/A) locus, the odds ratio for AG and AA genotypes versus GG genotypes were 2.31 (1.24-4.30; 95% confidence interval (CI)) and 0<0.001 (<0.001 to >999.999; 95% CI) which indicated that a proportion of AG homozygote was significantly higher in pTB-affected animals as compared with the control. General linear model analysis demonstrated higher MAP antibody response in TLR1 (+1380 AG) genotype as compared with TLR1 (+1380 GG) (P<0.0001). Present findings suggest that selection against TLR1 (+1380 G/A) may reduce the risk of pTB in bovine herds.

Y-chromosomal variation of local goat breeds of Turkey close to the domestication centre.

Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male-specific region markers, sex-determining region-Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honamli, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y-chromosomal variants have a more breed-dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.

INVESTIGATION OF STAT5A, FSHR AND LHR GENE POLYMORPHISMS IN TURKISH INDIGENOUS CATTLE BREEDS (EAST ANATOLLAN RED, SOUTH ANATOLIAN RED, TURKISH GREY, ANATOLIAN BLACK AND ZAVOT).

The objective of this study was to examine the allelic and genotypic profiles of the Signal Transducer and Activator of Transcription 5A (STAT5A), Follicle Stimulating Hormone Receptors (FSHR), and Luteinizing Hormone Receptor (LHR) genes in five indigenous cattle breeds in Turkey. For this purpose, a total of 329 cattle from East Anatolian Red (EAR), South Anatolian Red (SAR), Turkish Grey (TG), Anatolian Black (AB), and Zavot were genotyped using by PCR-RFLP method. A215 bp fragment of STAT5A, a 306bp fragment of FSHR, and a 303 bp fragment of LHR were amplified and digested with AvaI, AluI, and HhaI restriction enzymes, respectively. In this study two types of alleles C and Tfor STAT5A, C and G for FSHR and C and T for LHR were observed. The highest frequencies for STAT5A-C and STAT5A-T alleles were estimated for the Zavot and TG breeds (0.86) and the EAR breed (0.29), respectively. The highest frequency for FSHR-C and FSHR-G alleles was estimated for the Zavot breed (0.72) and the AB and SARbreeds (0.35), respectively. The highest frequency for LHR-C and LHR-T alleles was estimated for the EAR breed (0.75) and the AB breed (0.39), respectively. According to FT values, a small level of genetic diversity was found among five cattle breeds. The F(ST) value was calculated 0.019 between AB and Zavot. And, the value was significant (p < 0.001), while the other F(ST) values were not significant. According to the genetic distance values (Nei), the highest genetic distance was found between AB and TG while the smallest genetic distance was found between Zavot and TG. The chi-square test showed that the TG and Zavot breeds were in Hardy-Weinberg equilibrium (HWE) for STAT5A gene; the EAR, SAR, TG, and Zavot breeds were in HWE for FSHR gene and the EAR, SAR, and TG breeds were in HWE for LHR gene. In conclusion, further investigation is required to determine the correlation of the FSHR and LHR genes with early puberty for the improvement of sexual precocity, and it is considered that the STAT5A gene may be used to improve the milk yield and milk yield traits of local cattle breeds, including those indigenous to Turkey.

The effectiveness of gender determination using polymerase chain reaction and radioimmunoassay methods in cattle.

The aim of this study was to use polymerase chain reaction (PCR) by amplifying DNA from bovine (Bos taurus) fetal cells recovered through uterine puncture and subsequent amniotic fluid aspiration and to compare the effectiveness of the PCR method with amniotic dihydrotestosterone (DHT) levels in gender determination. Amniotic DHT levels between sexes were significantly higher in males than in females in all periods except the period 91 to 120 d. The differences among the amniotic DHT levels at different gestation periods (61 to 90, 91 to 120, 121 to 150, 151 to 180, 181 to 210 d) were not significant in females but were significant in males in the period 61 to 90 d compared with three other periods. Sensitivity was equal to 97.8% (95% CI=88.2% to 99.6%), and specificity was equal to 85.4% (95% CI=80.0% to 97.6%). These two values correspond with a cutoff of DHT in amniotic fluid. Distributions of the two sex groups were classified according to the 192.1 pg/mL cutoff value. A total of 93 amniotic fluid samples were examined by PCR analysis. The sex determination of 91 samples by PCR and electrophoresis was in agreement with the visual sexes of the fetuses. In two amniotic fluid samples, DNA was not isolated, and thus no sex determination was made. Fetal gender was correctly identified by PCR in 44 of 45 males and in 47 of 48 females. In PCR, one band (at the length of 102bp) and two bands (at the lengths of 102 and 226bp) were observed respectively for female and male fetuses. It may be concluded that the levels of amniotic DHT and PCR might be used for embryo sexing in pregnant cows.

Detection of bovine leukocyte adhesion deficiency (BLAD) in Turkish native and Holstein cattle.

The purpose of this work was to study whether the bovine leukocyte adhesion deficiency (BLAD) allele is present in native cattle breeds and the Holstein breed in Turkey. Blood samples were obtained from 120 Holstein, 20 Brown Swiss, 20 Anatolian Black, 20 Turkish Grey, 20 South Anatolian Red and 20 East Anatolian Red cattle. The isolated DNA materials were multiplied in PCR using the primer developed by Kriegesmann et al. (1997). In order to determine the area of mutation in PCR products, the PCR products were digested with TaqI endonuclease enzyme. The resulting fragments were analysed on 2% agarose gel for the absence of a TaqI restriction site. It was found that two of the Holstein cattle (a bull and a cow) were heterozygote BLAD carriers. There was no homozygote BLAD animal. The BLAD allele was not found in the other breeds used in the study. The mutant BLAD allele frequency in the 120 Holstein cattle calculations was 0.0084.