Atherosclerosis Induced by a High-Cholesterol and High-Fat Diet in the Inbred
Strain of the Wuzhishan Miniature Pig
Yuqiong Zhaoa, Lei Xiang a, Yaqian Liu a, Miaomiao Niu a, Jifang Yuan a and Hua Chen a,b aLaboratory Animal Center, Chinese PLA General Hospital, Beijing, PR China; bState Key Laboratory of Kidney Diseases, Chinese PLA General Hospital, Beijing, PR China
ABSTRACT
Coronary artery disease has a significant genetic predisposition, which mainly results from atherosclerosis. Miniature pig is an excellent model to investigate atherosclerosis. This study investigated whether the occurrence and development of atherosclerosis in the Wuzhishan miniature pigs (WZSPs) that were closely bred 12 generations had better consistency. The WZSPs (n ¼ 9) were fed a high-cholesterol and high-fat diet (HCFD). After continuous feeding, 3 WZSPs each were sacrificed at 6, 8, and 12 months, respectively, and the general clinical manifestations and serological indexes were detected. The pathological changes of the major arteries and main organs were recorded. The results showed WZSPs were quite susceptible to the HCFD. At 6 months, plaque lesions appeared in the abdominal aorta and iliac artery, while at 8 months, they appeared in the coronary artery. At 12 months, atherosclerotic lesions could be found in all major arteries, while lipid core, cholesterol precipitation, and calcium deposition appeared in the most serious sites. The progression of arterial lesions and distribution of the lesions were highly consistent in the
pigs. However, apparent variations in serum markers were observed. In conclusion, inbred WZSP is a good model to investigate atherosclerosis and has good predictability for the occurrence and development of the disease.
KEYWORDS
Atherosclerosis; inbreeding; miniature pig
Effect of Vitamin E and Polyunsaturated Fatty Acids on Cryopreserved Sperm
Quality in Bos taurus Bulls Under Testicular Heat Stress
João D. A. Losano, Daniel S. R. Angrimani, Andressa Dalmazzo, Carolina C. Rocha, Maíra M. Brito, Eduardo G. A. Perez, Roberta H. Tsunoda, Paola A. A. Góes, Camilla M. Mendes, Mayra E. O. A. Assumpção, Valquiria H. Barnabe and Marcilio Nichi
Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil
ABSTRACT
Taurine bulls are highly susceptible to heat stress, leading to increased oxidative stress (OS) and impaired sperm viability. Polyunsaturated fatty acids (PUFAs) supplementation can be an alternative to improve semen quality, which also results in more sperm susceptibility to lipid peroxidation. Moreover, this deleterious effect can be exacerbated in animals affected by heat stress. Vitamin E is a key antioxidant that counteracts lipid peroxidation of sperm membrane caused by OS. Thus, combining PUFAs with vitamin E may improve sperm quality. In this context, this study aimed to evaluate the effect of interaction between PUFAs and vitamin E on sperm quality in Bos taurus bulls under testicular heat stress. Sixteen taurine bulls under testicular heat stress were randomly assigned in four groups: Control, Vitamin E, PUFA, and PUFA þ Vitamin E. All groups lasted for 60 days. Samples were cryopreserved/thawed and analyzed for motility variables (CASA), membrane and acrosome integrity, mitochondrial activity, susceptibility to oxidative stress, DNA integrity, and sperm-binding capacity. Results showed that vitamin E had a beneficial effect on some sperm characteristics, whereas PUFA supplementation had an adverse effect when the two treatments were evaluated separately. Finally, the association between PUFAs and vitamin E did not improve sperm quality.
KEYWORDS
Bos taurus bulls; heat stress; oxidative stress; polyunsaturated fatty acids; semen; vitamin E
Effects of Recombinant Human Lactoferrin on Osteoblast Growth and Bone
Status in Piglets
Qiuling Li a,b, Jie Zhaoa, Wenping Hu a, Jianwu Wang c, Tian Yu c, Yunping Dai a and Ning Li a
aState Key Laboratory of AgroBiotechnology, China Agricultural University, Beijing, China; bCollege of Life Sciences, Langfang Teachers University, Langfang, China; cWuxi Kingenew Biotechnology Company, Wuxi, China
ABSTRACT
Lactoferrin (LF), an ~80 kDa iron-binding glycoprotein, modulates many biological effects, including antimicrobial and immunomodulatory activities. Recently, it was shown that LF also regulates bone cell activity, suggesting its therapeutic effect on postmenopausal bone loss. However, a minimal amount is known regarding the effects of recombinant human LF (rhLF)
supplementation on bone status in young healthy infants. We found osteoblast cell differentiation was significantly promoted in vitro. Furthermore, treatment of human osteoblast cells with rhLF rapidly induced phosphorylation of p44/p42 mitogen-activated protein kinase (p44/p42 MAPK, ERK1/2). In order to investigate the effects of rhLF on bone status in vivo, we used a piglet model, which is a useful model for human infants. Piglets were supplemented with rhLF milk for 30 days. Bone formation markers, Serum calcium concentration, bone mineral density (BMD), bone mineral content (BMC), tibia bone strength, and the overall metabolite profile analysis showed that rhLF was advantageous to the bone growth in piglets. These findings suggest that rhLF supplementation benefits neonate bone health by modulating bone formation.
KEYWORDS
Bone status; metabolites; osteoblast; piglets; recombinant human lactoferrin (rhLF)
Evaluation of Bovine High-Density SNP Genotyping Array in Indigenous Dairy
Cattle Breeds
S. Dash a, A. Singh a, A. K. Bhatia b, S. Jayakumarb, A. Sharmab, S. Singh b, I. Gangulyb and S. P. Dixitb
aICAR- National Dairy Research Institute, Karnal, Haryana, India; bICAR- National Bureau of Animal Genetic Resources, Karnal, Haryana, India
ABSTRACT
In total 52 samples of Sahiwal (19), Tharparkar (17), and Gir (16) were genotyped by using BovineHD SNP chip to analyze minor allele frequency (MAF), genetic diversity, and linkage disequilibrium among these cattle. The common SNPs of BovineHD and 54K SNP Chips were also extracted and evaluated for their performance. Only 40%50% SNPs of these arrays was found
informative for genetic analysis in these cattle breeds. The overall mean of MAF for SNPs of BovineHD SNPChip was 0.248 � 0.006, 0.241 � 0.007, and 0.242 � 0.009 in Sahiwal, Tharparkar and Gir, respectively, while that for 54K SNPs was on lower side. The average Reynold’s genetic distance between breeds ranged from 0.042 to 0.055 based on BovineHD Beadchip, and from 0.052 to 0.084 based on 54K SNP Chip. The estimates of genetic diversity based on HD and 54K chips were almost
same and, hence, low density chip seems to be good enough to decipher genetic diversity of these cattle breeds. The linkage disequilibrium started decaying (r2 < 0.2) at 140 kb inter-marker distance and, hence, a 20K low density customized SNP array from HD chip could be designed for genomic selection in these cattle else the 54K Bead Chip as such will be useful.
KEYWORDS
BovineHD BeadChip; genetic diversity; indigenous dairy cattle; linkage disequilibrium; minor allele frequency
Generation of Genomic Deletions (of Rig-I GENE) in Goat Primary Cell Culture
Using CRISPR/CAS9 Method
Shivani Malpotra, Ashutosh Vats, Sushil Kumar, Devika Gautam and Sachinandan De
Animal Genomics Lab, Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India
ABSTRACT
CRISPR/Cas9 system is a natural immune system in prokaryotes protecting them from infectious viral or plasmid DNA invading the cells. This RNA-guided system can act as powerful tool for introducing genomic alterations in eukaryotic cells with high efficiency. In the present study, Rig-Igene is taken as model gene to study the efficiency of CRISPR/Cas9 system induced gene deletion in primary fibroblast cell culture. Rig-I(retinoic acid-inducible gene-1) is involved in regulating immune response in mammals. In this study, we optimized the CRISPR/Cas9 method for knocking out Rig-Igene in Goat primary fibroblasts by using a NHEJ pathway. Cells were screened for inactivation of the Rig-Igene and two positive clones were found out of thirty colonies screened. Thus, cells containing Rig-Igene inactivation could be achieved by CRISPR/Cas9 in goat fibroblast cells.
KEYWORDS
Immunity; Genome editing; CRISPR/cas9; RIG-1
Evaluation of Bovine High-Density SNP Genotyping Array in Indigenous Dairy
Cattle Breeds
S. Dash a, A. Singh a, A. K. Bhatia b, S. Jayakumarb, A. Sharmab, S. Singh b, I. Gangulyb and S. P. Dixitb
aICAR- National Dairy Research Institute, Karnal, Haryana, India; bICAR- National Bureau of Animal Genetic Resources, Karnal, Haryana, India
ABSTRACT
In total 52 samples of Sahiwal (19), Tharparkar (17), and Gir (16) were genotyped by using BovineHD SNP chip to analyze minor allele frequency (MAF), genetic diversity, and linkage disequilibrium among these cattle. The common SNPs of BovineHD and 54K SNP Chips were also extracted and evaluated for their performance. Only 40%50% SNPs of these arrays was found
informative for genetic analysis in these cattle breeds. The overall mean of MAF for SNPs of BovineHD SNPChip was 0.248 � 0.006, 0.241 � 0.007, and 0.242 � 0.009 in Sahiwal, Tharparkar and Gir, respectively, while that for 54K SNPs was on lower side. The average Reynold’s genetic distance between breeds ranged from 0.042 to 0.055 based on BovineHD Beadchip, and from 0.052 to 0.084 based on 54K SNP Chip. The estimates of genetic diversity based on HD and 54K chips were almost
same and, hence, low density chip seems to be good enough to decipher genetic diversity of these cattle breeds. The linkage disequilibrium started decaying (r2 < 0.2) at 140 kb inter-marker distance and, hence, a 20K low density customized SNP array from HD chip could be designed for genomic selection in these cattle else the 54K Bead Chip as such will be useful.
KEYWORDS
BovineHD BeadChip; genetic diversity; indigenous dairy cattle; linkage disequilibrium; minor allele frequency
Generation of Genomic Deletions (of Rig-I GENE) in Goat Primary Cell Culture
Using CRISPR/CAS9 Method
Shivani Malpotra, Ashutosh Vats, Sushil Kumar, Devika Gautam and Sachinandan De
Animal Genomics Lab, Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India
ABSTRACT
CRISPR/Cas9 system is a natural immune system in prokaryotes protecting them from infectious viral or plasmid DNA invading the cells. This RNA-guided system can act as powerful tool for introducing genomic alterations in eukaryotic cells with high efficiency. In the present study, Rig-Igene is taken as model gene to study the efficiency of CRISPR/Cas9 system induced gene deletion in primary fibroblast cell culture. Rig-I(retinoic acid-inducible gene-1) is involved in regulating immune response in mammals. In this study, we optimized the CRISPR/Cas9 method for knocking out Rig-Igene in Goat primary fibroblasts by using a NHEJ pathway. Cells were screened for inactivation of the Rig-Igene and two positive clones were found out of thirty colonies screened. Thus, cells containing Rig-Igene inactivation could be achieved by CRISPR/Cas9 in goat fibroblast cells.
KEYWORDS
Immunity; Genome editing; CRISPR/cas9; RIG-1
Hepatoprotective, Hypoglycemic, and Hypolipidemic Effect of Chokeberry
Pomace on Polish Merino Lambs*
Paulina Lipińska and Artur Jóźwik
Institute of Genetics and Animal Breeding PAS in Jastrzębiec, Magdalenka near Warsaw, Poland
ABSTRACT
To properly understand the biological effect of polyphenol compounds, it should not be assumed that they are generally bioavailable and are reaching tissues as expected. It is important to fully understand how polyphenol compounds are absorbed, metabolized, and finally eliminated from the organism. Most polyphenol compounds occur in the forms that cannot be absorbed in their primary form. Sheep make compounds bioavailable due to their unique digestive system properties. Despite this, several species of ruminants likely to serve as an experimental model were considered. However, due to the high costs for cattle and low popularity in the region of goats, sheep were selected as the experimental model. In the study, 24 Polish Merino lambs were
randomly divided into a control and two experimental groups. The experimental groups were administered diets with the addition of 150 or 300 g of chokeberry pomace per each kg of complete feed mixture. Phenolics present in the chokeberry pomace were found to modulate biochemical blood parameters, resulting in hypoglycemic and hypolipidemic effects. Finally, the increase in total phenolics in the serum and liver of lambs induced a health-promoting effect on liver metabolic profile parameters.
KEYWORDS
Health-promoting effect; nutraceuticals; phenolics; ruminants
Mutations in Leptin (LEP) Gene Are Associated with Carcass and Meat Quality
Traits in Crossbreed Rabbits
Lukasz Migdal a, Konrad Koziol a, Sylwia Palka a, Wladyslaw Migdal b, Tomasz Zabekc,
Agnieszka Otwinowska-Mindura, Anna Migdal d, Michal Kmiecika, Dorota Maja and Jozef Bienieka
aDepartment of Genetics and Animal Breeding, Faculty of Animal Sciences, University of Agriculture in Krakow, Krakow, Poland; bDepartment Animal Product Technology, Faculty of Food Technology, University of Agriculture in Krakow, Krakow, Poland; cDepartmenof Genomics and Animal Molecular Biology, National Research Institute of Animal Production Poland; dInstitute of Veterinary Sciences, Faculty of Animal Sciences, University of Agriculture in Krakow, Krakow, Poland
ABSTRACT
Leptin is a hormone synthesized and secreted primarily in adipose cells that help to regulate energy balance. This study examined the associations of single nucleotide polymorphisms in the rabbit leptin gene with growth traits, slaughter traits and physicochemical parameters of New Zealand White (NZW) and Belgian Giant Grey (BGG) crossbreed rabbits. In total, 320 crossbreed animals were genotyped for polymorphisms within exon 2—g.16081633T>C, intron 1_2—g.16081420C>T, and
within UTR—g.16079636C>G for association analysis. Identified polymorphisms within rabbits leptin gene showed significant differences for dissectible fat percentage in carcass and dissectible fat weight in intermediate part (g.16081633T>C). Moreover, meat traits like protein content (g.16081633T>C; g.16079636C>G), intramuscular fat content (g.16081633T>C; g.16079636C>G,
g.16081420C>T), dry matter (g.16081420C>T), ash (g.16081420C>T), water (g.16081420C>T), and cohesiveness (g.16081420C>T, g.16079636C>G) were affected by polymorphisms in leptin gene. We conclude that polymorphism in the rabbit leptin gene influences important carcass and meat traits of NZW � BGG crossbreeds. Therefore, polymorphisms identified in this study may be used in selection as a meat trait markers.
KEYWORDS
Fatness traits; meat traits; rabbits; SNPs
Polymorphisms in MX2 Gene Are Related with SCS in Chinese Dairy Cows
Ningbo Chen, FengQiao Wang, Nongqi Yu, Yuan Gao, Jieping Huang, Ruihua Dang, Yongzhen Huang,
Xianyong Lan, Chuzhao Lei and Hong Chen
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China
ABSTRACT
Viral infections can play direct or indirect roles in the etiology of the bovine mastitis. Mx dynaminlike GTPase 2 (MX2) gene is a main effector of the antiviral innate immune defense mediated by type I interferon (IFN I), which was demonstrated to confer positive antiviral responses to many viruses. Given the importance of the MX2 in modulating the host immune response, MX2 gene may be a suitable candidate gene for studying disease resistance in dairy cattle. Here, we scanned the sequence variation of the MX2 gene in Chinese indigenous cattle breeds. Twenty-three previously reported SNPs were identified. To further analyze the effects of SNPs detected on mastitis disease, analysis of two SNPs (g.787527 C > T and g.787610 T > C) from 297 Chinese Holstein cows revealed a significant association with somatic cell score (SCS). Although functional studies are necessary to
ascertain whether these two SNPs are causal polymorphisms or merely in linkage with the true causal SNPs, implementation of these two SNPs as genetic markers in the dairy industry may be beneficial in selecting individuals with lower SCS.
KEYWORDS
50 UTR; marker assisted selection; mastitis; MX dynamin-like GTPase 2 (MX2); SNPs
Single Nucleotide Polymorphisms in 5’ Upstream Region of Bovine TLR4
Gene Affecting Expression Profile and Transcription Factor Binding Sites
Chinmoy Mishra, Subodh Kumar, Manjit Panigrahi, H. M. Yathish, Rajni Chaudhary, Anuj Chauhan, Amit Kumar and Arvind A. Sonawane
Department of Animal Genetics, Indian Veterinary Research Institute, Uttar Pradesh, India
ABSTRACT
The present study in the 5’ upstream region of TLR4 gene revealed four Single Nucleotide Polymorphisms (SNPs) in Vrindavani and Tharparkar cattle. The polymorphic information content (PIC), heterozygosity and allelic diversity values were low to moderate for these SNPs. In Vrindavani cattle, one SNP was found to be in Hardy-Weinberg Equilibrium (HWE) and the remaining three were found to be in linkage disequilibrium (LD) as indicated statistically (P > 0.05). In Tharparkar cattle, two SNPs were found to be in HWE and were not in LD as indicated statistically (P > 0.05). These SNPs were used for construction of haplotypes. In-silico analysis of these SNPs predicted abolition of eight transcription factor binding sites and creation of eight new sites. The quantitative real time PCR analysis did not show any significant variation of gene expression among haplotypes.
However, gene expression between breed was found to be significant (P < 0.05) which suggested that upstream region of bovine TLR4 gene has a crucial role in its expression. These findings in TLR4 gene offer essential evidence that can be useful in future research exploring its role in immunity. TLR4 can be used as a marker for selection for disease resistance in bovines.
KEYWORDS
Cattle; Haplotype; Linkage; SNP; TLR4
