Naturally Occurring Glycosidases in Milk from Native Cattle Breeds: Activity and Consequences on Free and Protein Bound-Glycans

Little is known about the extent of variation and activity of naturally occurring milk glycosidases and their potential to degrade milk glycans. A multi-omics approach was used to investigate the relationship between glycosidases and important bioactive compounds such as free oligosaccharides and O-linked glycans in bovine milk. Using 4-methylumbelliferone (4-MU) assays activities of eight indigenous glycosidases were determined, and by mass spectrometry and 1H NMR spectroscopy various substrates and metabolite products were quantified in a subset of milk samples from eight native North European cattle breeds. The results showed a clear variation in glycosidase activities among the native breeds. Interestingly, negative correlations between some glycosidases including β-galactosidase, N-acetyl-β-d-glucosaminidase, certain oligosaccharide isomers as well as O-linked glycans of κ-casein were revealed. Further, a positive correlation was found for free fucose content and α-fucosidase activity (r = 0.37, p-value < 0.001) indicating cleavage of fucosylated glycans in milk at room temperature. The results obtained suggest that milk glycosidases might partially degrade valuable glycans, which would result in lower recovery of glycans and thus represent a loss for the dairy ingredients industry if these activities are pronounced.

On the origin and diversification of Podolian cattle breeds: testing scenarios of European colonization using genome-wide SNP data

Background During the Neolithic expansion, cattle accompanied humans and spread from their domestication centres to colonize the ancient world. In addition, European cattle occasionally intermingled with both indicine cattle and local aurochs resulting in an exclusive pattern of genetic diversity. Among the most ancient European cattle are breeds that belong to the so-called Podolian trunk, the history of which is still not well established. Here, we used genome-wide single nucleotide polymorphism (SNP) data on 806 individuals belonging to 36 breeds to reconstruct the origin and diversification of Podolian cattle and to provide a reliable scenario of the European colonization, through an approximate Bayesian computation random forest (ABC-RF) approach. Results Our results indicate that European Podolian cattle display higher values of genetic diversity indices than both African taurine and Asian indicine breeds. Clustering analyses show that Podolian breeds share close genomic relationships, which suggests a likely common genetic ancestry. Among the simulated and tested scenarios of the colonization of Europe from taurine cattle, the greatest support was obtained for the model assuming at least two waves of diffusion. Time estimates are in line with an early migration from the domestication centre of non-Podolian taurine breeds followed by a secondary migration of Podolian breeds. The best fitting model also suggests that the Italian Podolian breeds are the result of admixture between different genomic pools. Conclusions This comprehensive dataset that includes most of the autochthonous cattle breeds belonging to the so-called Podolian trunk allowed us not only to shed light onto the origin and diversification of this group of cattle, but also to gain new insights into the diffusion of European cattle. The most well-supported scenario of colonization points to two main waves of migrations: with one that occurred alongside with the Neolithic human expansion and gave rise to the non-Podolian taurine breeds, and a more recent one that favoured the diffusion of European Podolian. In this process, we highlight the importance of both the Mediterranean and Danube routes in promoting European cattle colonization. Moreover, we identified admixture as a driver of diversification in Italy, which could represent a melting pot for Podolian cattle.

Association of SNPs in AKIRIN2, TTN, EDG1 and MYBPC1 genes with growth and carcass traits in Qinchuan cattle

Growth and carcass traits are the main breeding objectives in beef cattle. The aim of this study was to confirm genetic effects of the c.*188G>A SNP of AKIRIN2, the g.231054C>T SNP of TTN, the g.1471620G>T SNP of EDG1 and the g.70014208A>G SNP of MYBPC1 on growth and carcass traits in Chinese Qinchuan (QC) cattle, as well as to compare the frequencies of the well-characterized alleles of these SNPs among six Chinese cattle populations, three Japanese cattle populations, two European cattle populations and one Korean cattle population. In this study, a total of 665 cattle samples were genotyped using MassARRAY and PCR-RFLP. Association analysis explored effects of four SNPs on growth and carcass traits including body length, wither height, hip height, hip width, rump length, chest depth, chest circumference, back fat thickness, ultrasound longissimus muscle area and ultrasound longissimus muscle depth in QC (P<0.05 to P<0.001). The well-characterized A (c.*188G>A), T (g.231054C>T) and T (g.1471620G>T) alleles in Japanese Black cattle were significantly higher than Chinese cattle breeds, on the contrary, the G allele (g.70014208A>G) was markedly higher in Chinese cattle breeds than other cattle breeds. These results suggest that the four SNPs might be useful as a molecular marker for growth-related traits in Chinese QC cattle.

Association Study of Polymorphic Varients of Beta (B) Casein Gene with Milk Production Traits (Lactose, Snf and Density) in Malvi, Nimari, Sahiwal and H.F. Cross Breeds Cow

Background: The most frequently observed forms of beta casein (β-Cn) in dairy cattle are A1 and A2. A2 β-Cn is recognized as the original β-Cn protein because it existed before a mutation caused the appearance of A1 β-Cn in European cattle (Bos taurus) a few thousand years ago. In cattle, beta-casein (CSN2) gene is highly polymorphic with at least 13 genetic variants known until now.Methods: Research work was carried out on 50 Malvi and 50 Nimari, 50 Sahiwal and 50 H F Cross bred cow. In present research work the PCR amplicons of 121bp were digested by restriction endonuclease enzyme DdeI, which recognizes G^AATTC sites. Present association study of polymorphic variants’ showed that the presence of no restriction sites for the enzyme DdeI in both Malvi and Nimari. So one band of 121bp was observed on the gel and such genotype was designated as A2A2 type. Whereas in Sahiwal and H F Crossbred showed two type of genotypes A1A2 and A2A2.Result: The result of RFLP revealed that the gene and genotypic frequencies of β-casein (CSN2) gene for A2A2 was 1.00 for both Malvi and Nimari breed of cattle but 0.00, 0.30 and 0.70 in Sahiwal and 0.00, 0.64 and 0.36 in HF crossbred cattle, respectively. Association study showed that the lactose per cent was significantly higher in Nimari as compared to Malvi and Sahiwal. Among all the four breeds of cattle, significantly lower SNF (%) and density was noticed in Malvi breed of cattle for A2A2 genotype compared to remaining three breeds.

Polymorphism of the Melanocortin 1 Receptor (MC1R) Gene and its Role in Determining the Coat Colour of Central European Cattle Breeds

There are many genes responsible for the appearance of different coat colours, among which the melanocortin 1 receptor gene (MC1R) plays an important role. The aim of the study was to characterize genetic variation in Central European cattle breeds based on polymorphism of the MC1R gene and factors determining their coat colour. The study was conducted on 290 individuals of the following breeds: Polish White-Backed (PW), Lithuanian White-Backed (LW), Polish Red (PR), Lithuanian Red (LR), Carpathian Brown (CB), Ukrainian Grey (UG), and Slovak Pinzgau (SP). Polymorphism at the MC1R gene locus was analysed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using two restriction enzymes: Cfr10I and SsiI. The proportions of alleles and genotypes in the MC1R locus indicates a strong relationship between polymorphism and the coat colour of cattle: The ED allele proved to be characteristic for the breeds with a white-backed coat (PW and LW), while the dominant allele in the red breeds (PR and LR) was E+. It is noteworthy that coat colour in the SP population was determined only by the recessive e allele, which resulted in the formation of a separate clade in the phylogenetic tree.

SNPs in AKIRIN2, TTN, EDG1 and MYBPC1 genes are associated with growth-related traits in Chinese Qinchuan cattle

Growth performances are the main breeding objectives in Chinese beef cattle. The objective of this study was to confirm genetic effects of the c.*188G > A SNP in AKIRIN2, the g.231054C > T SNP in TTN, the g.1471620G > T SNP in EDG1, and the g.70014208A > G SNP in MYBPC1 gene on growth-related traits in Chinese Qinchuan cattle, as well as to compare the frequencies of the well-characterized alleles of these SNPs among six Chinese cattle populations, three Japanese cattle populations, two European cattle populations and one Korean cattle breed. In this study, a total of 655 cattle samples were genotyped using MassARRAY and PCR-RFLP. Association analysis indicated that the four SNPs have effects one to six indexes of growth-related traits including body length, wither height, hip height, hip width, rump length, chest depth and chest circumference in Chinese Qinchuan cattle (P < 0.05 to P < 0.001). The well-characterized A (c.*188G > A), T (g.231054C > T) and T (g.1471620G > T) alleles in Japanese Black cattle were significantly higher than Chinese cattle breeds, on the contrary, the G allele (g.70014208A > G) was markedly higher in Chinese cattle breeds than other cattle breeds. These results suggest that the four SNPs might be useful as a molecular marker for growth-related traits in Chinese Qinchuan cattle.