Body mass index and shortened telomere length in middle-aged female and male RUNNING HEAD: Middle-aged and shortened telomere length

السمنة هي عامل الخطر المرتبط بالاضطرابات المرتبطة بالعمر والتي تسرع الشيخوخة ،وتزيد من خطر الإصابة بأمراض الايض الغذائي. لذلك، أجريت هذه الدراسة للتحري عن ارتباط طول التيلومير في الكريات البيض (LTL) وزيادة وزن الجسم في النساء والرجال في منتصف العمر. شملت مجاميع الدراسة 160 (80 مجموعة سيطرة و80 مجموعة مؤشر كتلة الجسم عالي) مع أعمار تراوحت بين 30-50 سنة وتم مطابقتها لمؤشر كتلة الجسم. تم قياس التحاليل الفسلجية-الكيموحيوية باستخدام التحديد الأنزيمي. تم تحديد متوسط ​​طول التيلومير باستخدام تقنية وصمة سوذرن . أظهرت تحاليل الارتباط تباينًا معنويًا (P < 0.01) في المعايير الكيموحيوية بين مجاميع مؤشر كتلة الجسم الأعلى ومجموعة السيطرة بما في ذلك الخصر وصورة الدهون ومستوى الاستراديول والتستوستيرون والهرمون المحفز للجريبات والهرمون اللوتيني. كان متوسط ​​طول التيلومير أقصرعند الذكور في منتصف العمر منه في الإناث في مجاميع مؤشر كتلة الجسم الاعلى ومجاميع السيطرة لكلا المجموعتين العمريتين. كان LTL أقصر في المرضى الذين يعانون من زيادة الوزن والسمنة مقارنة مع مجموعة السيطرة، وكانت هذه الاختلافات في LTL لمجموعة السمنة أقصر من مجموعة الوزن الزائد. في الاستنتاج ، لوحظ قصر طول التيلومير في الرجال عند متوسط العمر الذين يعانون من زيادة الوزن و اضطراب الدهون. يمكن استخدام اضطراب الدهون / البروتين الدهني كمؤشر على قصر طول التيلومير ويمكن أن يؤدي تقليل مؤشرات السمنة إلى تحسين طول التيلومير في الأشخاص الذين يعانون من زيادة الوزن والسمنة.Obesity is a risk factor associated with age-related disorders that accelerate aging, and it increases the risk of metabolic diseases. Therefore, this study was conducted to investigate the association of leukocyte telomere length (LTL) with the presence of higher body weight in middle-aged females and males. The study subjects comprised 160 (80 control and 80 higher body mass index BMI groups) with ranging ages of 30-50 years included and stratified for BMI. The physio-biochemical analysis was measured using enzymatic determination. Mean telomere length was determined by using the southern blotting technique. The association analysis revealed a significant variance (P < 0.01) in biochemical parameters between higher BMI groups and control including waist, lipid profile, and the level of estradiol, testosterone, follicle-stimulating hormone, and luteinizing hormone. Mean telomere length was shorter in middle-aged males compared to the females of higher BMI groups and control groups for both age groups. LTL was shorter in the overweight and obese patients compared with the control group, and these differences in LTL obese group were shorter compared to the overweight group. In conclusion, shorter telomere length was observed in middle-aged males associated with higher body weight and lipid abnormalities. Lipid/lipoprotein abnormalities can be used as a predictor for the shortened telomere length and the reduction in adiposity indices can improvement the telomere length in both overweight and obese subjects

Influence of litter size on the hematologic profile of Awassi ewes during gestation and lactation

Physiological status and litter size affect ewes' hematological parameters, which serve as health indicators. These parameters reflect changes in an animal's physiological condition. Therefore, this study examined the hematological profiles of Awassi ewes with single and twin pregnancies during pregnancy and postpartum. A total of 232 sexually mature and healthy ewes (123 with single pregnancies and 109 with twin pregnancies) between the ages of 3 and 4 years were included in the current study. The sheep's blood was collected and hematological tests were conducted immediately on the samples. Results revealed variation in hematological parameters between ewes during pregnancy based on litter size. Twin-pregnant ewes had significantly higher amounts of RBC, PCV %, MCH, and MCHC during pregnancy months as compared to single-pregnant ewes. Compared to single pregnancies, Awassi ewes with twin pregnancies had higher leukocytes, lymphocytes, granulocytes, lymphocytes %, and granulocytes % than those with a singleton. Logistic regression confirmed that litter size contributes to the increase in hematological parameters. In conclusion, Awassi ewes with twin pregnancies exhibited an increase in the counts of leukocytes and erythrocytes constituents. This study provided valuable information about the association of twin-pregnant ewes with hematological parameters that could be used in sheep breeding and reproduction to improve diagnosis, prognosis, and treatment in the field

Genotyping of Diacylglycerol Acyltransferase 2 Gene in Holstein Cattle Population

This study was conducted to describe the single nucleotide polymorphism (SNPs) within exon 8 and its intron 9 flanking region within bovine Diacylglycerol Acyltransferase 2 (DGAT2) gene and the possible association of these SNPs with the milk productive traits. Blood samples were collected from randomly selected Holstein cattle. Genomic DNA was extracted, and a pair of specific PCR primers was designed to amplify a segment that consists of partial exon 8/partial intron 9 of DGAT2 gene. SSCP experiments were optimized and performed for each amplified PCR fragment. Each set of SSCP resolved bands was sequenced and analyzed. Four SSCP patterns representing four genotypes (BD, BB, CD, and AD) were detected with four alleles. Several novel genetic polymorphisms were discovered. Three SNPs (157 C/A, 158 T/G, and 159 G/A) were found in genotype BD,  while one SNP (94 G/T,  153 C/A, 154 T/A) were found in the genotypes BB, CD, and AD, respectively. The only non-synonymous SNP was found in genotype CD (344 D/Y), and the effect of this missense mutation on the protein three-dimensional structure was visualized. The sequence homology between the bovine DGAT2 gene and other species was also analyzed. The significance of the correlation of each genotype with the productive traits of milk was observed. The Holstein cattle with BD genotype produced significantly more milk for all studied 90 days of lactation, with a significant effect on fat for only the last 30 days of lactation, whereas there was no effect on protein and lactose percentage for the entire lactation period. The results suggest a novel association between bovine DGAT2 genetic variability and the milk yield in Holstein cattle. This opens interesting prospects for future DGAT2 based selection programs and preservation strategies

Association of Kappa casein gene polymorphism with milk production traits in crossbred dairy cows

Milk's qualitative and technological properties are greatly affected by genetic polymorphisms in the kappa-casein gene, and their polymorphisms may serve as informative markers of yield and composition. Thus, the objective of this study was to detect kappa-casein (kappa-CN) gene polymorphisms and their association with milk production traits in crossbred dairy cows. One hundred healthy crossbred (Friesian x Jenoubi) dairy animals between three and five years old were sampled for blood and milk during their first lactation. The genomic DNA was extracted from whole blood, and restriction fragment length polymorphism (RFLP-PCR) was used to determine the genotype of the kappa-CN gene. As a consequence of the restriction digestion of this fragment with Hind III, it showed three different restriction patterns: BB (453 base pairs uncut), AB (453, 206, and 225 base pairs), and AA (206 and 225 base pairs). Based on genetic diversity, the AB genotype was the most predominant (n = 67), with a frequency of 0.67. A variant genotype of the kappa-CN gene was associated with milk production traits in crossbred dairy cows. Animals with the AA variant produced a higher milk yield and a higher percentage of fat, casein, protein, and solids not fat (SNF) (P≤0.05) (1.397kg, 0.75%, 0.31%, 0.27%, and 0.68%, respectively) than those with the BB variant. A logistic regression analysis confirmed that the kappa-CN genotypes increase milk yield and casein content. Therefore, genetic variants of the kappa-CN gene could be used as genetic markers for improving milk production traits in dairy cattle. Keywords: cattle, genetic variants, milk protein

Genotyping of Diacylglycerol Acyltransferase 2 Gene in Holstein Cattle Population

This study was conducted to describe the single nucleotide polymorphism (SNPs) within exon 8 and its intron 9 flanking region within bovine Diacylglycerol Acyltransferase 2 (DGAT2) gene and the possible association of these SNPs with the milk productive traits. Blood samples were collected from randomly selected Holstein cattle. Genomic DNA was extracted, and a pair of specific PCR primers was designed to amplify a segment that consists of partial exon 8/partial intron 9 of DGAT2 gene. SSCP experiments were optimized and performed for each amplified PCR fragment. Each set of SSCP resolved bands was sequenced and analyzed. Four SSCP patterns representing four genotypes (BD, BB, CD, and AD) were detected with four alleles. Several novel genetic polymorphisms were discovered. Three SNPs (157 C/A, 158 T/G, and 159 G/A) were found in genotype BD,  while one SNP (94 G/T,  153 C/A, 154 T/A) were found in the genotypes BB, CD, and AD, respectively. The only non-synonymous SNP was found in genotype CD (344 D/Y), and the effect of this missense mutation on the protein three-dimensional structure was visualized. The sequence homology between the bovine DGAT2 gene and other species was also analyzed. The significance of the correlation of each genotype with the productive traits of milk was observed. The Holstein cattle with BD genotype produced significantly more milk for all studied 90 days of lactation, with a significant effect on fat for only the last 30 days of lactation, whereas there was no effect on protein and lactose percentage for the entire lactation period. The results suggest a novel association between bovine DGAT2 genetic variability and the milk yield in Holstein cattle. This opens interesting prospects for future DGAT2 based selection programs and preservation strategies

Sex Determination of Unknown Ovine Samples Gathered from Slaughters

Accurate and specific methods for sex determination in farm animals play a vital role in animal resources. Therefore, this study was conducted to differentiate between males and females for unknown sheep samples collected from slaughters based on the amelogenin (AMELX/ AMELY) gene by using the polymerase chain reaction (PCR) technique. A total of 200 blood samples of Awassi sheep, collected from slaughterhouses, with an average age ranging between 3-6 years. Blood samples from the sheep were used to isolate the DNA. Specific PCR primers were used to amplify one fragment (262 bp) from the X- chromosome in ewes, and two fragments (262 and 202 bp) from the X and Y- chromosome in rams, respectively. The specificity of the primers was evaluated by using amelogenin amplicons for samples of known sex. The results revealed a single band for ewes (262 bp) and two bands for the rams (202 and 262 bp) after electrophoresis. In conclusion, this assay is proved to be a precise, inexpensive, and favorable method in sex determination, especially when unknown samples are collected from the slaughterhouse, and the samples are mixed so that it is difficult for the researcher to determine the sex of the samples. This method can be employed in other vertebrates and sexing of offspring in animal breeding because the amelogenin gene is conserved among mammals. [J Bangladesh Agril Univ 2022; 20(4.000): 402-406

Fatty Acid Synthase Polymorphism of Awassi Sheep and its Impact on Fatty Acid Composition

Livestock intramuscular fat content is determined by fatty acid synthase (FASN).  The fatty acid profile of the fat is of relevance to human health.  Thus, this study explores the relationship between the polymorphism of the FASN gene in Awassi sheep and its impact on fatty acid composition.  The study used 100 Awassi rams, ranging in age from 1 to 2.5 years.  Molecular DNA was isolated from each blood sample; genotyping, sequencing reactions, and in silico tools were subsequently used to confirm the variants in amplified fragments.  The results revealed two genotypes (GG and GA) of the ovine FASN gene (exon 2).  The novel missense c.186 GA was identified in the genotype GA.  The GA genotype had significantly (P0.05) increased intramuscular fat, higher unsaturated fatty acid content, and lower saturated fatty acid content than the GG genotype.  Cumulative in silico analysis indicated a detrimental effect of the SNP c.186 GA on FASN activity.  The intramuscular GA genotype of Awassi sheep had a low saturated content relative to unsaturated fats.  The result confirmed that the c.186 GA SNP in ovine FASN gene variation is potentially helpful in assessing carcass traits, and this might lead to a more efficient selection of animals with healthier fatty acid profiles, resulting in higher-quality meat

A Missense p.Q>R234 Mutation in the Osteopontin Gene Is Associated With the Prolificacy of Iraqi Awassi Ewes

One of the most valuable traits in production and breeding is a sheep’s prolificacy which is influenced by several genes, one of which is the osteopontin ( OPN ) gene. Thus, this study aimed to determine the effect of genetic variation within the OPN gene on Awassi ewe prolificacy. Genomic DNA was extracted from 123 single-progeny ewes and 109 twin ewes. Polymerase chain reaction (PCR) was used to amplify 4 sequence fragments (289, 275, 338, and 372 bp), representing exons 4, 5, 6, and 7 of the OPN gene. A 372 bp amplicon was identified with 3 different genotypes: TT, TC, and CC. Sequence analysis revealed a novel mutation in TC genotypes p.Q>R234. Statistical analysis revealed that the single nucleotide polymorphism (SNP) p.Q>R234 was associated with prolificacy. Ewes carrying the p.Q>R234 SNP had significantly ( P  ⩽ .01) lower litter sizes, twinning rates, and lambing rates, and more days to lambing than those with the TC and TT genotypes. The p.Q>R234 SNP was confirmed to be responsible for lower litter size through logistic regression analysis. From these results, we can conclude that the missense variant p.Q>R234 adversely affects the traits of interest and shows that the p.Q>R234 SNP negatively influences the prolificacy of Awassi sheep. Based on this study, it is evident that ewes in this population carrying the p.Q>R234 SNP have a lower litter size and are less prolific

GHRL gene-based genotyping of ovine and caprine breeds reveals highly polymorphic intronic sequences in Awassi sheep with several RNA motifs

Abstract Background The current study was conducted to identify the genetic polymorphism of ghrelin (GHRL) gene of sheep and goats, as well as to determine whether these polymorphisms were associated with the evolutionary genetic differences in the involved species. This study was performed on 233 sheep and 91 goats. Two genetic loci of 113 bp and 262 bp partially spanning over exon 2/intron 2 and intron 4/exon 5 of GHRL gene respectively were amplified and genotyped using polymerase chain reaction–single-strand conformation polymorphism (PCR-SSCP) and DNA sequencing methods. Results The SSCP banding pattern of 262-bp locus indicated the presence of four diplotypes (BC, BB, AC, and AB) in Awassi sheep, three diplotypes (BC, BB, and AB) in Karadi sheep, and only two diplotypes (BC and BB) in all goats’ samples. The current study detected several novel SNPs in the ovine–caprine populations as well as two SNPs that are observed only in sheep, including intron4:119 C>A and intron4:123 T>G. The phylogenetic analysis revealed that the observed diplotypes resided within ovine sequences and were closely related to caprine counterparts. Computational analyses indicated the presence of various intronic RNA motifs. However, all these motifs were gathered in Awassi breed. Conclusion It is stated that the intron 4 is highly diverse amongst goats and sheep as well as within sheep with a particular emphasis on Awassi. This genetic peculiarity may in turn suggest a high polymorphic pattern of this breed in comparison with other related counterparts