1.Effects of the ITPR1 gene overexpression on Ca²⁺ concentration, lipid content and calcium transport-related genes in duck uterine epithelial cells.
Minfang YOU ; Yuanyu QIN ; Yiyu ZHANG ; Chaomei LIAO ; Guanghui TAN ; Jiezhang LI ; Wangui LI
Chinese Journal of Biotechnology 2021;37(7):2443-2452
Inositol 1,4,5-trisphosphate receptor 1 (ITPR1) is an important intracellular channel for releasing Ca²⁺. In order to investigate the effects of the ITPR1 overexpression on Ca²⁺ concentration and lipid content in duck uterine epithelial cells and its effects on calcium transport-related genes, the structural domain of ITPR1 gene of duck was cloned into an eukaryotic expression vector and transfected into duck uterine epithelial cells. The overexpression of the ITPR1 gene, the concentration of Ca²⁺, the lipid content, and the expression of other 6 calcium transport-related genes was determined. The results showed that the concentration of Ca²⁺ in uterine epithelial cells was significantly reduced after transfection (P<0.05), the triglyceride content was significantly increased (P<0.01), and the high-density lipoprotein content was significantly decreased (P<0.01). The correlation analysis results showed that the overexpression of the C-terminal half of the ITPR1 gene was significantly positively correlated with the total cholesterol content (P<0.01), which was significantly positively correlated with the low-density lipoprotein content (P<0.05). The overexpression of the N-terminal half of the ITPR1 gene was significantly positively correlated with the triglyceride content (P<0.01), which was significantly negatively correlated with the concentration of Ca²⁺ (P<0.05). RT-qPCR results of 6 calcium transport-related genes showed that the overexpression of the C-terminal half of the ITPR1 gene significantly inhibited the expression of the IP3R2, VDAC2 and CAV1 genes, and the overexpression of the N-terminal half of the ITPR1 gene significantly promoted the expression of the IP3R3 and CACNA2D1 genes. In conclusion, the ITPR1 gene overexpression can promote Ca²⁺ release in duck uterus epithelial cells, promote the synthesis of triglyceride, low-density lipoprotein and cholesterol, and inhibit the production of high-density lipoprotein, and the ITPR1 gene overexpression affected the expression of all 6 calcium transport-related genes.
Animals
;
Calcium/metabolism*
;
Ducks/genetics*
;
Epithelial Cells
;
Female
;
Inositol
;
Inositol 1,4,5-Trisphosphate Receptors
;
Lipids
;
Uterus
2.Effects of SCD-1 gene overexpression on the content of calcium ion and lipids in duck uterine epithelial cells.
Jiezhang LI ; Hualun LUO ; Guanghui TAN ; Lei WU ; Yuanyu QIN ; Yiyu ZHANG
Chinese Journal of Biotechnology 2020;36(5):899-907
Stearoyl-CoAdesaturase-1 (SCD-1) is a key regulator of monounsaturated fatty acid synthesis. It plays a vital role in lipid synthesis and metabolism. Ca²⁺ is an important cation in the body and plays an important role in the organism. The aims of this study were to investigate the correlation of SCD-1 gene overexpression with lipid indexes and calcium ion level. The pcDNA3.1 (+) + SCD-1 +Flag eukaryotic expression vector and cultured duck uterine epithelial cells were co-transfected. The overexpression of SCD-1 gene was measured using the Flag Label Detection Kit. Ca ions and lipid contents were detected through Fluo-3/AM Calcium Ion Fluorescence Labeling method and Lipid Measuring Kit, respectively. SCD-1 gene overexpression was negatively correlated with triglyceride (TG) and high-density lipoprotein cholesterol (HDL-C), and positively correlated with Ca ion, total cholesterol (TC), very low-density lipoprotein cholesterol (VLDL-C) and low density lipoprotein cholesterol (LDL-C) levels. Meanwhile, Ca ion was positively correlated with TG, LDL-C and HDL-C contents, and negatively correlated with TC and VLDL-C levels. Overexpression of SCD-1 gene could regulate Ca ion secretion, as well as lipid synthesis and transport in duck uterine epithelial cells.
Animals
;
Calcium
;
metabolism
;
Coenzyme A Ligases
;
genetics
;
Ducks
;
Epithelial Cells
;
chemistry
;
enzymology
;
Gene Expression
;
Ions
;
Lipids
;
genetics
;
Triglycerides
;
metabolism
3.Cloning and expression of duck C4BPα and verification of its interaction with Riemerella anatipestifer.
Delong LI ; Lijuan TAN ; Jiulong GU ; Siyuan WANG ; Ting LIU ; Sihuai CHEN ; Jiye GAO ; Fashu TANG ; Jixiang LI
Chinese Journal of Biotechnology 2020;36(4):693-699
To study the interaction between C4b-binding protein (C4BP) and Riemerella anatipestifer (RA), we cloned duck C4BPα, conducted prokaryotic expression and prepared the polyclonal antibody by immunizing mice. Then indirect immunofluorescence assay and dot blotting hybridization assay were used to verify the interaction between C4BP and RA. The full length of duck C4BPα nucleotide sequence was 1 230 bp, with the highest similarity to chicken C4BPα (82.1%). Phylogenetic tree analysis showed that duck C4BPα and chicken C4BPα were on the same phylogenetic tree branch and the genetic evolution relationship between them was the closest. C4BPα was efficiently expressed in Escherichia coli BL21 (DE3). The recombinant proteins existed in intracellular soluble form. The titer of polyclonal antibody was more than 1:10 000 and polyclonal antibodies could specifically recognize the recombinant proteins. The results of indirect immunofluorescence assay and dot blot hybridization assay showed that RA could interact with duck C4BP. The results provide a basis to further reveal the pathogenesis of RA.
Animals
;
Cloning, Molecular
;
Complement C4b-Binding Protein
;
chemistry
;
genetics
;
metabolism
;
Ducks
;
classification
;
genetics
;
microbiology
;
Gene Expression Regulation
;
Mice
;
Phylogeny
;
Riemerella
;
metabolism
4.Multiple food allergy – unexpected culprits
Cristina Isabel de Figueiredo ORNELAS ; Fátima CABRAL DUARTE ; Maria Conceição Galvão PEREIRA DOS SANTOS ; Manuel Augusto de Castro PEREIRA BARBOSA
Asia Pacific Allergy 2018;8(3):e30-
Food allergy has an estimated prevalence of 6%–8% in children. Meat allergy and multiple food allergy due to sensitization to cross-reactive components in infancy is, however, less frequent. A 5-year-old girl was referred to our department with a multiple food allergy history. She had severe immediate worsening of her atopic dermatitis with hen's egg (6 months) and cow's milk introduction (7 months). At the age of 9 months, she presented with recurrent and reproducible atopic dermatitis' worsening and lip edema with the introduction of different meats (chicken, turkey, cow, pork, and rabbit), having the same complaints with fish at 12 months (salmon and hake). At her first appointment she was avoiding hen's egg, cow's milk, meat, and fish (except fresh tuna, codfish, and pollock). We performed skin prick tests (commercial extract and prick-to-prick with whole food) and specific IgE, which revealed sensitization to hen's egg, raw meat (cow, pork, chicken, turkey, duck, lamb, goat, and rabbit; negative for cooked meat), codfish and cow's milk (mild). ISAC was performed, revealing sensitization to 3 cross-reactive components (serum albumins Bosd6, Canf3, and Feld2) and specific food components of chicken's egg/meat (Gald1, 2, 3, and 5), cod (Gadc1), hazelnut (Cora9), and kiwi (Actd1). We present a rare case of multiple food allergy in infancy, where sensitization to cross-reactive components was responsible for most of the children complaints. The detection of serum albumins' involvement was especially important, because it can possibly mean tolerance to these foods in well-cooked forms, substantially improving patient and family's quality of life.
Albumins
;
Chickens
;
Child
;
Child, Preschool
;
Corylus
;
Dermatitis, Atopic
;
Ducks
;
Edema
;
Female
;
Food Hypersensitivity
;
Goats
;
Humans
;
Hypersensitivity
;
Immunoglobulin E
;
Lip
;
Meat
;
Milk
;
Ovum
;
Prevalence
;
Quality of Life
;
Red Meat
;
Serum Albumin
;
Skin
;
Tuna
;
Turkey
5.Molecular Genetic Findings of Spirometra decipiens and S. ranarum in Korea
Hyeong Kyu JEON ; Sun HUH ; Woon Mok SOHN ; Jong Yil CHAI ; Keeseon S EOM
The Korean Journal of Parasitology 2018;56(4):359-364
The taxonomy of Spirometra species has been controversial despite the medical and veterinary importance. Currently, only a few Spirometra species are considered valid species in the genus Spirometra. In the present study, the distribution of Spirometra species obtained from animals in Korea were identified by molecular analysis of the mitochondrial cytochrome c oxidase I (cox1) gene. A total of 28 Spirometra species specimens were analyzed. These were all collected between 1973 and 2008 in the Republic of Korea. Mitochondrial cox1 sequences were examined for a total of 28 specimens comprising 14 S. decipiens and 14 S. ranarum. The difference in partial cox1 sequences (316 bp) between S. erinaceieuropaei (KJ599680) and S. ranarum (this study) was 9.3%, while that between S. decipiens (KJ599679) and S. ranarum (this study) was 2.2%. Genetic analyses identified 2 Spirometra species in animals such as cat, leopard cat, dog, duck and snake in Korea as S. decipiens and S. ranarum. S. decipiens and S. ranarum were present in Gyeongnam Province (P), Jeonnam P, Gangwon P, Chungbuk P, and Seoul. S. decipiens was found in tadpoles, snakes, ducks, cats, leopard cats and dogs, while S. ranarum was found in cats and dogs. The ratio of S. decipiens:S. ranarum calculated from the molecular data was 14:14 (or 1:1). These results indicate that S. decipiens and S. ranarum are sympatrically distributed in Korea.
Animals
;
Cats
;
Chungcheongbuk-do
;
Classification
;
Dogs
;
Ducks
;
Electron Transport Complex IV
;
Gangwon-do
;
Jeollanam-do
;
Korea
;
Larva
;
Molecular Biology
;
Panthera
;
Republic of Korea
;
Seoul
;
Snakes
;
Spirometra
6.A two-component signal transduction system contributes to the virulence of Riemerella anatipestifer
Qing WANG ; Mianmian CHEN ; Wei ZHANG
Journal of Veterinary Science 2018;19(2):260-270
Similar to other studies of bacterial pathogens, current studies of the pathogenesis of Riemerella anatipestifer (RA) are focused mainly on in vitro culture conditions. To elucidate further the pathogenesis of RA in vivo, bacterial RNA was extracted from overnight tryptic soy broth cultures (in vitro) and from the blood of infected ducks (in vivo) for comparative RNA sequencing analysis. In total, 682 upregulated genes were identified in vivo. Among the upregulated genes, a signal transduction response regulator (ArsR) and a signal transduction histidine kinase (SthK) were predicted to be located on the same operon. A mutant was constructed by deletion of both of these genes. Duck infection tests showed that genes ArsR and SthK were related to the virulence of the pathogen in vivo. Differentially expressed genes identified by comparison of in vitro and in vivo conditions provided an insight into the physiological process of RA infection and provided an opportunity to identify additional virulence factors.
Ducks
;
Genes, vif
;
Histidine
;
In Vitro Techniques
;
Operon
;
Phosphotransferases
;
Physiological Processes
;
Riemerella
;
RNA, Bacterial
;
Sequence Analysis, RNA
;
Signal Transduction
;
Virulence Factors
;
Virulence
7.Novel reassortants of clade 2.3.4.4 H5N6 highly pathogenic avian influenza viruses possessing genetic heterogeneity in South Korea in late 2017
Yu Na LEE ; Sun Ha CHEON ; Soo Jeong KYE ; Eun Kyoung LEE ; Mingeun SAGONG ; Gyeong Beom HEO ; Yong Myung KANG ; Hyun Kyu CHO ; Yong Joo KIM ; Hyun Mi KANG ; Myoung Heon LEE ; Youn Jeong LEE
Journal of Veterinary Science 2018;19(6):850-854
Novel H5N6 highly pathogenic avian influenza viruses (HPAIVs) were isolated from duck farms and migratory bird habitats in South Korea in November to December 2017. Genetic analysis demonstrated that at least two genotypes of H5N6 were generated through reassortment between clade 2.3.4.4 H5N8 HPAIVs and Eurasian low pathogenic avian influenza virus in migratory birds in late 2017, suggesting frequent reassortment of clade 2.3.4.4 H5 HPAIVs and highlighting the need for systematic surveillance in Eurasian breeding grounds.
Agriculture
;
Animals
;
Birds
;
Breeding
;
Ducks
;
Ecosystem
;
Genetic Heterogeneity
;
Genotype
;
Influenza in Birds
;
Korea
8.Genomic analysis of Sheldrake origin goose hemorrhagic polyomavirus, China
Chunhe WAN ; Cuiteng CHEN ; Longfei CHENG ; Rongchang LIU ; Guanghua FU ; Shaohua SHI ; Hongmei CHEN ; Qiuling FU ; Yu HUANG
Journal of Veterinary Science 2018;19(6):782-787
Goose hemorrhagic polyomavirus (GHPV) is not a naturally occurring infection in geese in China; however, GHPV infection has been identified in Pekin ducks, a domestic duck species. Herein, we investigated the prevalence of GHPV in five domestic duck species (Liancheng white ducks, Putian black ducks, Shan Sheldrake, Shaoxing duck, and Jinyun Sheldrake) in China. We determined that the Jinyun Sheldrake duck species could be infected by GHPV with no clinical signs, whereas no infection was identified in the other four duck species. We sequenced the complete genome of the Jinyun Sheldrake origin GHPV. Genomic data comparison suggested that GHPVs share a conserved genomic structure, regardless of the host (duck or geese) or region (Asia or Europe). Jinyun Sheldrake origin GHPV genomic characterization and epidemiological studies will increase our understanding of potential heterologous reservoirs of GHPV.
China
;
Ducks
;
Epidemiologic Studies
;
Geese
;
Genome
;
Polyomavirus
;
Prevalence
9.Alterations in antioxidant function and cell apoptosis in duck spleen exposed to molybdenum and/or cadmium.
Mengmeng ZHANG ; Junrong LUO ; Caiying ZHANG ; Huabin CAO ; Bing XIA ; Guoliang HU
Journal of Veterinary Science 2017;18(2):193-200
To investigate the effects of molybdenum (Mo) and/or cadmium (Cd) on antioxidant function and the apoptosis-related genes in duck spleens. Sixty healthy 11-day-old ducks were randomly divided into six groups of 10 ducks (control, low Mo group, high Mo, Cd, low Mo + Cd, and high Mo + Cd groups). All were fed a basal diet containing low or high dietary doses of Mo and/or Cd. Relative spleen weight, antioxidant indices, apoptosis-related gene mRNA expression levels, and ultrastructural changes were evaluated after 120 days. The results showed that the relative spleen weight decreased significantly in the high Mo + Cd treatment group which compared with control group. Malondialdehyde levels increased and xanthine oxidase and catalase activities decreased in the Mo and/or Cd groups compared with levels in the control group. Bak-1 and Caspase-3 expressions were upregulated in the high Mo + Cd group, while Bcl-2 was downregulated. In addition, mitochondrial crest fracture, swelling, vacuolation, deformed nuclei, and karyopyknosis in both Mo + Cd treated groups were more severe than in the other groups. The results suggest that Mo and/or Cd can induce oxidative stress and apoptosis of spleen via effects on the mitochondrial intrinsic pathway. Moreover, the results indicate the two elements have a possible synergistic relationship.
Apoptosis*
;
Cadmium*
;
Caspase 3
;
Catalase
;
Diet
;
Ducks*
;
Malondialdehyde
;
Molybdenum*
;
Oxidative Stress
;
RNA, Messenger
;
Spleen*
;
Xanthine Oxidase
10.Identification of microbiome with 16S rRNA gene pyrosequencing and antimicrobial effect of egg white in bovine mastitis.
Danil KIM ; Eun Kyung KIM ; Won Jin SEONG ; Younghye RO ; Dae Sung KO ; Nam Hyung KIM ; Jae Hong KIM ; Hyuk Joon KWON
Korean Journal of Veterinary Research 2017;57(2):117-126
Bovine mastitis is an important microbial disease in the dairy industry. We investigated the frequencies of bacterial pathogens in 62 farms and pathogen antibiotic resistance from mastitis samples (n = 748). We tested the antimicrobial activity of chicken and duck egg white and lysozyme purified from chicken egg white. Moreover, we compared the microbiomes of normal and mastitic raw milk obtained by 16S rRNA gene pyrosequencing and culture methods. The results showed that the frequencies of Gram-positive pathogens (Enterococcus faecalis 37% and Staphylococcus aureus 36%) were higher than that of a Gram-negative pathogen (Escherichia coli 15%). Resistance frequencies to ampicillin and norfloxacin were lowest in Staphylococcus aureus (21%), Enterococcus faecalis (23%), and Escherichia coli (33%), and the antimicrobial activity of chicken egg white was higher than those of lysozyme and duck egg white. Pyrosequencing results revealed clear differences between the microbiomes of mastitic and normal raw milk samples and revealed a slightly similar, but clearly different, composition of pathogens compared to that from the culture method. Thus, pyrosequencing may be useful for elucidating changes in microbiomes during mastitis progression and treatment. A chicken egg white and antibiotic combination may help with mastitis treatment; however, further studies are needed.
Agriculture
;
Ampicillin
;
Animals
;
Cattle
;
Chickens
;
Drug Resistance, Microbial
;
Ducks
;
Egg White*
;
Enterococcus faecalis
;
Escherichia coli
;
Female
;
Genes, rRNA*
;
Mastitis
;
Mastitis, Bovine*
;
Methods
;
Microbiota*
;
Milk
;
Muramidase
;
Norfloxacin
;
Ovum*
;
Staphylococcus aureus

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