Polymorphism detection of <i>DGAT1</i> and <i>Lep</i> genes in Anatolian water buffalo (<i>Bubalus bubalis</i>) populations in Turkey

Acyl-CoA: diacylglycerol–acyltransferase 1 (DGAT1) enzyme plays a key role in controlling the synthesis rate triglyceride from diacylglycerol. Leptin (LP, OB, obese) is an important hormone that synthesizes mostly from adipose tissue and regulates glucose metabolism and homeostasis. DGAT1 and Lep genes are closely related to reproduction, growth, milk yield and composition in water buffalo breeds. This study aimed to identify genetic variation in the DGAT1 and Lep gene regions in 150 water buffalo individuals from five different provinces of Turkey using DNA sequencing. A total of 38 nucleotide variations and indels have identified 761 bp long partial intron 2 and exon 3 and 5′ UTR regions of the Lep gene in Anatolian water buffalo populations; 422 bp long partial exon 7–9 and exon 8 regions of DGAT1 gene were amplified and two mutations were defined in the point of 155 and 275 nucleotide that is three genotypes for S allele and Y allele of DGAT1 gene in intron 7 in Anatolian buffalo populations, respectively. These SNPs may have an effect on reproduction, growth, milk yield and composition in water buffalo populations and may prove to be useful for water buffalo breeding.

MOLECULAR GENODIAGNOSTICS OF PRODUCER BULLS BY ECONOMICALLY VALUABLE CHARACTERISTICS

Based on the results of the analysis of the relationship between the DGAT1 gene polymorphism and economically valuable traits, it was concluded that animals with the DGAT1KK genotype have the highest milk yield and fat content. When analyzing the relationship of CSN3 gene polymorphism with milk productivity, it was found that higher rates are characteristic of mothers cows with a homozygous CSN3BB genotype. Animals of this group have an advantage, in comparison with the AB and AA genotypes, in milk yield, protein content, mass yield of fat and protein. When studying the data of the relationship between the LGB gene polymorphism and economically valuable traits, it was concluded that animals with the homozygous LGBBB genotype have the highest milk yield, fat, protein content, mass fat and protein yield.

The role of diglyceride acyltransferase (DGAT) genes polymorphisms on milk production in dairy goats

Milk quantity, fat, and protein contents are important traits in dairy animals, particu-larly in dairy goats. However, little is known about the genetics and genomics in dairy goats that influence these traits. The diacylglycerol O-acyltransferase (DGAT) genes are considered the most important genes in dairy goats for milk quantity and fat con-tents. The DGAT genes have two isoforms as DGAT1 and DGAT2, that have similar re-actions but no sequence homology in-between. The DGAT1 gene in chromosome 14 is a functional and potential candidate gene for fat content in goats, while DGAT2 is associated with milk yield and fat percentage in goats. The phylogenetic analyses re-vealed similar topologies and evolutionary partitioning of DGAT family proteins into two major clades: DGAT1 and DGAT2. Moreover, DGAT1 proteins were found to play a significant role in the milk yield in dairy goats. In this review, the role of polymor-phisms in DGAT family genes on milk production in dairy goats is discussed

DGAT1 GENE POLYMORPHISM AND ITS RELATIONSHIPS WITH CATTLE MILK YIELD AND CHEMICAL COMPOSITION

Several polymorphisms in different gene loci have been noted to affect production traits such as milk yield and milk composition. The present study aimed to determine the allelic and genotypic frequency of the DGAT1 gene and evaluate the associations between DGAT1 genetic variants and milk yield and its chemical composition of local Iraqi cattle. Blood samples from 100 cows were obtained for DNA isolation. The primer used in this study amplified 411-bp fragments at exon 8 of the DGAT1 gene. DNA sequencing methods were applied to detect single nucleotide polymorphism of the DGAT1 gene in 100 cows. The nucleotide sequences of exon 8 of the DGAT1 gene were registered for local Iraqi cattle in the National Center for Biotechnology Information (NCBI), DNA Data Bank of Japan (DDBJ), and the European Nucleotide Archive (ENA) under the following accession numbers (LC492073 and LC492074). The results showed the presence of two polymorphic sites leading to the construction of 2 different haplotypes in the cow. Haplotype diversity was 0.536, while nucleotide diversity was 0.0031. Two single-nucleotide polymorphism (SNP) loci of the DGAT1 gene were detected, namely A10433G (A/G) and A10434C (A/C). The resulting of this mutation changes lysine to alanine substitution at position 232 (A232K mutation) of amino acid sequence. Geneious software V. 2020.0.4 was used to detect genotypes of the DGAT1 gene, as the sequence alignment showed the presence of three genotypes. The genotypic frequencies of KK, KA, and AA were 0.40, 0.30, and 0.30, respectively. Frequencies of K and A alleles were 0.60 and 0.40, respectively. The KK genotype was significantly (P 0.05) associated with higher fat yield. Therefore, the DGAT1 gene could serve as a genetic marker for the selection of fat yield in cows.

Impact of SNPs in ACACA, SCD1, and DGAT1 Genes on Fatty Acid Profile in Bovine Milk with Regard to Lactation Phases

Milk fat is a dietary source of fatty acids (FA), which can be health promoting or can increase risks of some diseases. FA profile composition depends on many factors, among them gene polymorphism. This study analyzed the relation between polymorphism of acetyl-CoA carboxylase α (ACACA), stearoyl-CoA desaturase 1 (SCD1), diacylglycerol acyltransferase 1 (DGAT1) genes with FA profile in milk from Polish Holstein-Friesian cattle and determined changes of FA percentage during lactation with regard to polymorphism. Milk samples were collected twice: during the first phase of lactation (<90 Days in milk; DIM) and at the end of lactation (>210 DIM). During the first milk collection, blood samples were taken to analyze three chosen single nucleotide polymorphisms (SNPs): AJ312201.1g.1488C > G SNP in ACACA gene, A293V SNP in SCD1 gene, and K232A SNP in DGAT1 gene. Increased concentration of FA that are less beneficial for human health and have lower concentration of healthy FA in homozygotes: GG in ACACA, VV in SCD1, and KK in DGAT1 were observed, as well as a strong influence of the analyzed genes on FA with 18C atoms was also found. Moreover, it was demonstrated that lactation phase significantly affected FA percentage in milk depending on the phenotype. These results may contribute their part to knowledge toward obtaining more beneficial milk composition.

MILK OF BLACK-WHITE COWS WITH DIFFERENT GENES OF CAPPA CASEIN AND DIACYLGLYCEROL OF O-ACYLTRANSFERASE FOR CHEESEMAKING

The purpose of the research is improving the quality and technological properties of milk from black-and-white cows. The research was conducted on first-calf cows of a black-and-white breed in LLC «Dusym» of the Atninsky district of the Republic of Tatarstan. Three groups of experimental cows were formed depending on the genotype of CSN3 and DGAT1, which have equal lactation month. Cheese making milk obtained from cows with different allelic variants of the kappa-casein and diacylglycerol O-acyltransferase genes was studied. DNA testing was performed on the CSN3 and DGAT1 genes of experimental cows and allelic variants of these genes were determined by pol-ymerase chain reaction followed by analysis of restriction-site polymorphism. It was found that milk of cows with the genotype CSN3 AB and CSN3 BB, when exposed to rennet, showed a better yield of a dense casein clot (80 and 100%) and separation of serum (P <0.05), with a shorter coagulation time (20.3 and 16.5 min; P <0.05-0.01). Ac-cording to the DGAT1 gene, first-calf milk with the allelic variant DGAT1 AK and DGAT1 KK had the advantage, respectively, 86.7% and 85.7%, 24.0 and 25.7 minutes. The worst milk coagulation time is typical for CSN3 AA group – 29.5 minutes and DGAT1 AA one – 30.4 min. In terms of milk coagulation time, type II (15-40 min.) groups have the genotype CSN3 AB and CSN3 BB with the largest share of cows in the herd – 80.0 and 71.4% and with the genotype DGAT1 AK and DGAT1 KK – 86.6 and 85.7%, respectively. Thus, milk obtained from cows with the kappa-casein gene allele B and the diacylglycerol O-acyltransferase gene allele K gene in the genotype has the best cheese-making properties.

A110 DGAT1 MUTATION OF LIPID TRAFFICKING ASSOCIATED WITH A RARE CASE OF CONGENITAL DIARRHEA

Background Congenital diarrheas and enteropathies (CODEs) are rare and cause severe chronic diarrhea in children under 2 years of age.1 Current genomic research has identified several monogenic defects resulting in epithelial disorders of nutrient and electrolyte transport, enzymes function, metabolism and trafficking. Aims To discuss a rare case of congential diarrhea. Methods Case Report. Results A 10-month-old Ashkenazi Jewish boy was referred to Pediatric Gastroenterology for loose stools and failure to thrive. Born at 27 weeks, he was a NICU inpatient for his first 3 months of life. Explosive loose stools were first noticed after discharge and treated as cow’s milk protein allergy. At aged 10 months he was admitted to his local hospital with irritability, abdominal distension, foul smelling loose stools and developmental delay. Despite an intake of 130kcal/kg/day he failed to gain weight (weight and length &lt; 3rd centile). His abdomen was distended but there was no organomegaly on examination. On transfer to Sick Kids he underwent extensive work up with low albumin 24 g/L but normal CBC, amylase, lipase and IgA and anti-TTG negative. The lipid profile showed hypertriglyceridemia 2.98 mmol/L (normal &lt;0.85), low HDL 0.81 mmol/L but normal total cholesterol. Low apoliprotein A1 0.79 g/L (0.92 - 1.96), low apoliprotein B 0.54 g (0.59 - 1.46) and vitamin E deficiency 9.6 umol/L (14.5 - 33.0). Normal fecal elastase (&gt; 500 ug/g stool). Upper GI endoscopy showed duodenal villi blunting with non-specific chronic inactive duodenitis on histopathology. Due to ongoing poor weight gain he commenced TPN. A repeat endoscopy four weeks after admission revealed patchy villous blunting, crypt hyperplasia and enterocyte vacuolization. However, ultrastructural electron microscopy examination showed normal enterocyte brush border with well-formed microvilli with no inclusions or vesicular bodies. With a differential diagnosis of chylomicron retention disorder, abetalipoproteinemia or hypolipoproteinemia he was commenced on a low-fat diet with MCT feeds. A 72-hour fecal fat collection on this diet was within normal range 5.4 mmol/day (2–7). One year later his weight is now on 3rd centile and his albumin levels have normalized. However, triglycerides remain elevated. Whole exome sequencing identified two pathogenic variants in the DGAT1 gene; c.629_631delCCT, pSer210del and c751 + 2T&gt;C, IVSB+2T&gt;C. Conclusions Diacylglycerol acyltransferase (DGAT)1 catalyzes triglyceride biosynthesis.2 Loss of function DGAT1 gene mutations are described in several cases studies3,4 characterized by electrolyte transport–related diarrhea, protein-losing enteropathy and growth failure. Our cases adds to the understanding of the phenotypic and histological spectrum of DGAT1 mutations. Management strategies currently focus on growth with dietary lipid restriction, while substituting fat-soluble vitamins and essential fatty acids. Funding Agencies CAG