Imprinting at the KBTBD6 locus involves species-specific m ternal methylation and monoallelic expression in livestock animals

Background The primary differentially methylated regions(DMRs) which are maternally hypermethylated serve as imprinting control regions(ICRs) that drive monoallelic gene expression, and these ICRs have been investigated due to their implications in mammalian development. Although a subset of genes has been identified as imprinted, in-depth comparative approach needs to be developed for identification of species-specific imprinted genes. Here, we examined DNA methylation status and allelic expression at the KBTBD6 locus across species and tissues and explored potential mechanisms of imprinting.Results Using whole-genome bisulfite sequencing and RNA-sequencing on parthenogenetic and normal porcine embryos, we identified a maternally hypermethylated DMR between the embryos at the KBTBD6 promoter Cp G island and paternal monoallelic expression of KBTBD6. Also, in analyzed domesticated mammals but not in humans, non-human primates and mice, the KBTBD6 promoter Cp G islands were methylated in oocytes and/or allelically methyl-ated in tissues, and monoallelic KBTBD6 expression was observed, indicating livestock-specific imprinting. Further analysis revealed that these Cp G islands were embedded within transcripts in porcine and bovine oocytes which coexisted with an active transcription mark and DNA methylation, implying the presence of transcription-dependent imprinting.Conclusions In this study, our comparative approach revealed an imprinted expression of the KBTBD6 gene in domesticated mammals, but not in humans, non-human primates, and mice which implicates species-specific evolution of genomic imprinting.

Effects of arginine replacement with L-citrulline on the arginine/nitric oxide metabolism in chickens:An animal model without urea cycle

Background This study examined the efficacy of L-citrulline supplementation on the arginine/nitric oxide metabolism,and intestinal functions of broilers during arginine deficiency.A total of 288 day-old Arbor Acre broilers were randomly assigned to either an arginine deficient basal diet(NC diet),NC diet+0.50%L-arginine(PC diet),or NC diet+0.50%L-citrulline(NCL diet).Production performance was recorded,and at 21 days old,chickens were euthanized for tissue collection.Results The dietary treatments did not affect the growth performance of broilers(P>0.05),although NC diet increased the plasma alanine aminotransferase,urate,and several amino acids,except arginine(P<0.05).In contrast,NCL diet elevated the arginine and ornithine concentration higher than NC diet,and it increased the plasma citrulline greater than the PC diet(P<0.05).The nitric oxide concentration in the kidney and liver tissues,along with the plasma and liver e NOS activities were promoted by NCL diet higher than PC diet(P<0.05).In the liver,the activities of arginase 1,ASS,and ASL,as well as,the gene expression of i NOS and OTC were induced by PC diet greater than NC diet(P<0.05).In the kidney,the arginase 1,ASS and ASL enzymes were also increased by PC diet significantly higher than the NC and NCL diets.Comparatively,the kidney had higher abundance of n NOS,ASS,ARG2,and OTC genes than the liver tissue(P<0.05).In addition,NCL diet upregulated(P<0.05)the m RNA expression of intestinal nutrient transporters(EAAT3 and PEPT1),tight junction proteins(Claudin 1 and Occludin),and intestinal mucosal defense(MUC2 and p Ig R).The intestinal morphology revealed that both PC and NCL diets improved(P<0.05)the ileal VH/CD ratio and the jejunal VH and VH/CD ratio compared to the NC fed broilers.Conclusion This study revealed that NCL diet supported arginine metabolism,nitric oxide synthesis,and promoted the intestinal function of broilers.Thus,L-citrulline may serve as a partial arginine replacement in broiler’s diet without detrimental impacts on the performance,arginine metabolism and gut health of chickens.

Management practices and technologies for efficient biological sample collection from domestic animals with special reference to Indian field conditions

Management strategies and the use of advanced technologies are equally important for determining the sample number and sampling frame for successful field sampling for animal disease prevalence studies.The quality of the biological samples collected in the field has a direct bearing on the integrity of the data generated,prevalence estimates and subsequent policy decisions on disease control.Hence,compromising the quality of biological samples collected in the field could potentially undermine the priority setting principles in disease control strategies.Biological samples collected from domestic animals in the field are precious materials and require meticulous planning for sample collection,sample storage in the field,transportation,and storage in the laboratory.Poorly managed field sampling has a significant detrimental impact on the sample quality and quantity and directly affects the accuracy of disease prevalence data.A bad choice of sampling tools,containers,storage and transport all have a negative impact on the integrity of the sample and consequently have an impact on the outcome.Over the last two years,as part of our one health animal sampling work in ndia,we have observed challenges and opportunities in the field sampling of animals for disease prevalence studies.This paper aims to provide information on management practices and technologies for efficient biological sample collection from the field and ensure that good quality samples are available fortesting.

Prevotella and succinate treatments altered gut microbiota,increased laying performance,and suppressed hepatic lipid accumulation in laying hens

Background This work aimed to investigate the potential benefits of administering Prevotella and its primary metabolite succinate on performance,hepatic lipid accumulation and gut microbiota in laying hens.Results One hundred and fifty 58-week-old Hyline Brown laying hens,with laying rate below 80%and plasma triglyceride(TG)exceeding 5 mmol/L,were used in this study.The hens were randomly allocated into 5 groups and subjected to one of the following treatments:fed with a basal diet(negative control,NC),oral gavage of 3 mL/hen saline every other day(positive control,PC),gavage of 3 mL/hen Prevotella melaninogenica(10^(7)CFU/mL,PM)or 3 mL/hen Prevotella copri(10^(7)CFU/mL,P.copri)every other day,and basal diet supplemented with 0.25%sodium succinate(Succinate).The results showed that PM and P.copri treatments significantly improved laying rate compared to the PC(P<0.05).The amount of lipid droplet was notably decreased by PM,P.copri,and Succinate treatments at week 4 and decreased by P.copri at week 8(P<0.05).Correspondingly,the plasma TG level in Succinate group was lower than that of PC(P<0.05).Hepatic TG content,however,was not significantly influenced at week 4 and 8(P>0.05).PM treatment increased(P<0.05)the mRNA levels of genes PGC-1βand APB-5B at week 4,and ACC and CPT-1 at week 8.The results indicated enhanced antioxidant activities at week 8,as evidenced by reduced hepatic malondialdehyde(MDA)level and improved antioxidant enzymes activities in PM and Succinate groups(P<0.05).Supplementing with Prevotella or succinate can alter the cecal microbiota.Specifically,the abundance of Prevotella in the Succinate group was significantly higher than that in the other 4 groups at the family and genus levels(P<0.05).Conclusions Oral intake of Prevotella and dietary supplementation of succinate can ameliorate lipid metabolism of laying hens.The beneficial effect of Prevotella is consistent across different species.The finding highlights that succinate,the primary metabolite of Prevotella,represents a more feasible feed additive for alleviating fatty liver in laying hens.

Spatially resolved transcriptomic profiling of placental development in dairy cow

The placenta plays a crucial role in successful mammalian reproduction.Ruminant animals possess a semi-invasive placenta characterized by a highly vascularized structure formed by maternal endometrial caruncles and fetal placental cotyledons,essential for full-term fetal development.The cow placenta harbors at least two trophoblast cell populations:uninucleate(UNC)and binucleate(BNC)cells.However,the limited capacity to elucidate the transcriptomic dynamics of the placental natural environment has resulted in a poor understanding of both the molecular and cellular interactions between trophoblast cells and niches,and the molecular mechanisms governing trophoblast differentiation and functionalization.To fill this knowledge gap,we employed Stereo-seq to map spatial gene expression patterns at near single-cell resolution in the cow placenta at 90 and 130 days of gestation,attaining high-resolution,spatially resolved gene expression profiles.Based on clustering and cell marker gene expression analyses,key transcription factors,including YBX1 and NPAS2,were shown to regulate the heterogeneity of trophoblast cell subpopulations.Cell communication and trajectory analysis provided a framework for understanding cell-cell interactions and the differentiation of trophoblasts into BNCs in the placental microenvironment.Differential analysis of cell trajectories identified a set of genes involved in regulation of trophoblast differentiation.Additionally,spatial modules and co-variant genes that help shape specific tissue structures were identified.Together,these findings provide foundational insights into important biological pathways critical to the placental development and function in cows.

Mammary stem cells:expansion and animal productivity

Identification and characterization of mammary stem cells and progenitor cells from dairy animals is important in the understanding of mammogenesis, tissue turnover, lactation persistency and regenerative therapy. It has been realized by many investigators that altered lactation, long dry periods （non-milking period between two consecutive lactation cycles）, abrupt cessation of lactation （common in water buffaloes） and disease conditions like mastitis, greatly reduce milk yield thus render huge financial losses within the dairy sector. Cellular manipulation of specialized cell types within the mammary gland, called mammary stem cells （MaSCs）/progenitor cells, might provide potential solutions to these problems and may improve milk production. In addition, MaSCs/progenitor cells could be used in regenerative therapy against tissue damage caused by mastitis. This review discusses methods of MaSC/progenitor cell manipulation and their mechanisms in bovine and caprine animals. Author believes that intervention of MaSCs/progenitor cells could lessen the huge financial losses to the dairy industry globally.

Design of live-attenuated animal vaccines based on pseudorabies virus platform

Pseudorabies virus(PRV)is a double-stranded DNA virus with a genome approximating 150 kb in size.PRV contains many non-essential genes that can be replaced with genes encoding heterogenous antigens without affecting viral propagation.With the ability to induce cellular,humoral and mucosal immune responses in the host,PRV is considered to be an ideal and potential live vector for generation of animal vaccines.In this review,we summarize the advances in attenuated recombinant PRVs and design of PRV-based live vaccines as well as the challenge of vaccine application.

Breed identification using breed‑informative SNPs and machine learning based on whole genome sequence data and SNP chip data

Background Breed identification is useful in a variety of biological contexts.Breed identification usually involves two stages,i.e.,detection of breed-informative SNPs and breed assignment.For both stages,there are several methods proposed.However,what is the optimal combination of these methods remain unclear.In this study,using the whole genome sequence data available for 13 cattle breeds from Run 8 of the 1,000 Bull Genomes Project,we compared the combinations of three methods(Delta,FST,and In)for breed-informative SNP detection and five machine learning methods(KNN,SVM,RF,NB,and ANN)for breed assignment with respect to different reference population sizes and difference numbers of most breed-informative SNPs.In addition,we evaluated the accuracy of breed identification using SNP chip data of different densities.Results We found that all combinations performed quite well with identification accuracies over 95%in all scenarios.However,there was no combination which performed the best and robust across all scenarios.We proposed to inte-grate the three breed-informative detection methods,named DFI,and integrate the three machine learning methods,KNN,SVM,and RF,named KSR.We found that the combination of these two integrated methods outperformed the other combinations with accuracies over 99%in most cases and was very robust in all scenarios.The accuracies from using SNP chip data were only slightly lower than that from using sequence data in most cases.Conclusions The current study showed that the combination of DFI and KSR was the optimal strategy.Using sequence data resulted in higher accuracies than using chip data in most cases.However,the differences were gener-ally small.In view of the cost of genotyping,using chip data is also a good option for breed identification.

Sulforaphane prevents LPS‑induced inflammation by regulating the Nrf2‑mediated autophagy pathway in goat mammary epithelial cells and a mouse model of mastitis

Background Mastitis not only deteriorates the composition or quality of milk,but also damages the health and pro-ductivity of dairy goats.Sulforaphane(SFN)is a phytochemical isothiocyanate compound with various pharmacologi-cal effects such as anti-oxidant and anti-inflammatory.However,the effect of SFN on mastitis has yet to be elucidated.This study aimed to explore the anti-oxidant and anti-inflammatory effects and potential molecular mechanisms of SFN in lipopolysaccharide(LPS)-induced primary goat mammary epithelial cells(GMECs)and a mouse model of mastitis.Results In vitro,SFN downregulated the mRNA expression of inflammatory factors(tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-6),inhibited the protein expression of inflammatory mediators(cyclooxygenase-2(COX2),and inducible nitric oxide synthase(iNOS))while suppressing nuclear factor kappa-B(NF-κB)activation in LPS-induced GMECs.Additionally,SFN exhibited an antioxidant effect by increasing Nrf2 expression and nuclear translocation,up-regulating antioxidant enzymes expression,and decreasing LPS-induced reactive oxygen species(ROS)produc-tion in GMECs.Furthermore,SFN pretreatment promoted the autophagy pathway,which was dependent on the increased Nrf2 level,and contributed significantly to the improved LPS-induced oxidative stress and inflammatory response.In vivo,SFN effectively alleviated histopathological lesions,suppressed the expression of inflammatory factors,enhanced immunohistochemistry staining of Nrf2,and amplified of LC3 puncta LPS-induced mastitis in mice.Mechanically,the in vitro and in vivo study showed that the anti-inflammatory and anti-oxidative stress effects of SFN were mediated by the Nrf2-mediated autophagy pathway in GMECs and a mouse model of mastitis.Conclusions These results indicate that the natural compound SFN has a preventive effect on LPS-induced inflam-mation through by regulating the Nrf2-mediated autophagy pathway in primary goat mammary epithelial cells and a mouse model of mastitis,which may improve prevention strategies for mastitis in dairy goats.

State of the art in research on the gut-liver and gut-brain axis in poultry

The relationship between the intestines and their microbiota,the liver,and the neuronal system is called the gutliver-brain axis.This relationship has been studied and observed for a relatively short time but is considered in the development of research focused on,e.g.,liver diseases and intestinal dysbiosis.The role of the gut microbiota in this relationship is crucial,as it acts on poultry’s performance and feed utilization,affecting meat and egg quality.The correct composition of the intestinal microbiota makes it possible to determine the essential metabolic pathways and biological processes of the individual components of the microbiota,allowing further speculation of the role of microbial populations on internal organs such as the liver and brain in the organism.The gut microbiota forms a complex,dense axis with the autonomic and enteric nervous systems.The symbiotic relationship between the liver and gut microbiota is based on immune,metabolic and neuroendocrine regulation,and stabilization.On the other hand,the gut-brain axis is a bidirectional interaction and information transfer system between the gastrointestinal tract and the central nervous system.The following paper will discuss the current state of knowledge of the gut-liver-brain axis of poultry,including factors that may affect this complex relationship.

Phylogenetic and epidemiological characteristics of H9N2 avian influenza viruses in Shandong Province, China from 2019 to 2021

H9N2 avian influenza virus(AIV) has widely circulated in poultry worldwide and sporadic infections in humans and mammals. During our surveillance of chicken from 2019 to 2021 in Shandong Province, China, we isolated 11 H9N2AIVs. Phylogenetic analyses showed that the eight gene segments of the 11 isolates were closely related to several sublineages of Eurasian lineage: BJ/94-like clades(HA and NA genes), G1-like clades(PB2 and M genes), and SH/F/98-like clades(PB1, PA, NP and NS genes). The isolates showed mutation sites that preferentially bind to humanlike receptors(HA) and mammalian fitness sites(PB2, PB1 and PA), as well as mutations in antigen and drug resistance sites. Moreover, studies with mice revealed four isolates with varying levels of pathogenicity. The average antibody titer of the H9N2 AIVs was 8.60 log2. Based on our results, the epidemiological surveillance of H9N2 AIVs should be strengthened.

MED4 gene positively affects preadipocyte differentiation in Chinese red steppe cattle

Background:The regulatory genes related to lipid metabolism affect the intramuscular fat(IMF)content and improve meat quality traits.Mediator Complex Subunit 4(MED4),a vitamin D receptor protein,affects vitamin D,livestock growth,carcass traits,and triglyceride deposition.However,the physiological function of the MED4 gene on bovine adipocyte differentiation remains unknown.Methods:This study explored the function of the MED4 gene in preadipocyte differentiation in Chinese Red Steppe cattle.The overexpression plasmid and the interference sequences of the MED4 gene were constructed to detect the effects of the MED4 gene on adipogenesis and biomakers using quantitative polymerase chain reaction and western blotting.Results:The MED4 gene had significantly high expression during preadipocyte differentiation(p<0.05).Overexpression of the MED4 gene increased the expression of the PPARγgene,a preadipocyte differentiation biomarker,improved cellular lipid droplets and triglycerides accumulation,and positively accelerated adipocyte maturation(p<0.05).Interference of the MED4 gene can negatively regulate preadipocyte differentiation.Conclusion:This study showed that the MED4 gene may affect the preadipocyte differentiation and adipogenesis in Chinese Red Steppe cattle by regulating the PPARγgene.

High-efficiency Embryo Cryopreservation Technology for Mutton Sheep

Embryo cryopreservation is an important part of embryo biotechnology.It plays a key role in the development of quality animal husbandry.However,the effect of in-vivo(in-vitro)embryo freezing of cattle and sheep in China has not reached an expected level,which limits the commercial application of embryo transfer technology.The technical bottleneck that needs to be broken through is that embryos are subject to mechanical damage and protective agent toxicity during freezing and thawing.This paper summarized the successful application of plant antifreeze protein in vitrification preservation of pig embryos,which makes this technology widely used in animal husbandry production,and is of great significance in accelerating the large-scale breeding of improved livestock and saving endangered animals.

Genome-wide association study for numbers of vertebrae in Dezhou donkey population reveals new candidate genes

Numbers of vertebrae is an important economic trait associated with body size and meat productivity in animals.However,the genetic basis of vertebrae number in donkey remains to be well understood.The aim of this study was to identify candidate genes affecting the number of thoracic(TVn)and the number of lumbar vertebrae(LVn)in Dezhou donkey.A genome-wide association study was conducted using whole genome sequence data imputed from low-coverage genome sequencing.For TVn,we identified 38 genome-wide significant and 64 suggestive SNPs,which relate to 7 genes(NLGN1,DCC,SLC26A7,TOX,WNT7A,LOC123286078,and LOC123280142).For LVn,we identified 9 genome-wide significant and 38 suggestive SNPs,which relate to 8 genes(GABBR2,FBXO4,LOC123277146,LOC123277359,BMP7,B3GAT1,EML2,and LRP5).The genes involve in the Wnt and TGF-βsignaling pathways and may play an important role in embryonic development or bone formation and could be good candidate genes for TVn and LVn.

The effect of ensiled paulownia leaves in a high-forage diet on ruminal fermentation,methane production,fatty acid composition,and milk production performance of dairy cows

Background:The use of industrial by-products rich in bioactive compounds as animal feeds can reduce greenhouse gas production.Paulownia leaves silage(PLS)was supplemented to dairy cows'diet and evaluated in vitro(Exp.1;Rusitec)and in vivo(Exp.2,cannulated lactating dairy cows and Exp.3,non-cannulated lactating dairy cows).The study investigated the PLS effect on ruminal fermentation,microbial populations,methane production and concentration,dry matter intake(DMI),and fatty acid(FA)proportions in ruminal fluid and milk.Results:Several variables of the ruminal fluid were changed in response to the inclusion of PLS.In Exp.1,the p H increased linearly and quadratically,whereas ammonia and total volatile fatty acid(VFA)concentrations increased linearly and cubically.A linear,quadratic,and cubical decrease in methane concentration was observed with increasing dose of the PLS.Exp.2 revealed an increase in ruminal p H and ammonia concentrations,but no changes in total VFA concentration.Inclusion of PLS increased ruminal propionate(at 3 h and 6 h after feeding),isovalerate,and valerate concentrations.Addition of PLS also affected several populations of the analyzed microorganisms.The abundances of protozoa and bacteria were increased,whereas the abundance of archaea were decreased by PLS.Methane production decreased by 11%and 14%in PLS-fed cows compared to the control in Exp.2 and 3,respectively.Exp.3 revealed a reduction in the milk protein and lactose yield in the PLS-fed cows,but no effect on DMI and energy corrected milk yield.Also,the PLS diet affected the ruminal biohydrogenation process with an increased proportions of C18:3 cis-9 cis-12 cis-15,conjugated linoleic acid,C18:1 trans-11 FA,polyunsaturated fatty acids(PUFA),and reduced n6/n3 ratio and saturated fatty acids(SFA)proportion in milk.The relative transcript abundances of the 5 of 6 analyzed genes regulating FA metabolism increased.Conclusions:The dietary PLS replacing the alfalfa silage at 60 g/kg diet can reduce the methane emission and improve milk quality with greater proportions of PUFA,including conjugated linoleic acid,and C18:1 trans-11 along with reduction of SFA.

Assessment of the Quality of Frozen-Thawed Semen or Epididymal Sperm in Three Native Vietnamese Pig Breeds

Our aim was to evaluate the quality of ejaculated and epididymal frozen-thawed pig sperm of endangered Vietnam native pig breeds. Ejaculated sperm was collected from live boars and epididymal sperm was collected from slaughtered boars of the MuongTe, Kieng Sat and Co BinhThuan breeds and frozen in 0.25 ml straws using a protocol established earlier for modern pig breeds. We evaluated the sperm quality after thawing in terms of motility and rates of viable and abnormal spermatozoa. Our results revealed that the sperm motility and rates of viable and abnormal frozen-thawed sperm were >30%, >44%, and <14%, respectively. The origin of sperm had an effect on the production of pig embryos in vitro. In the Co BinhThuan breed, ejaculated sperm generated higher cleavage, blastocyst and hatching rates than did the epididymal sperm (60.11% vs 56.02%, 17.23% vs 14.31%, 3.78% vs 2.34%, respectively, P < 0.05). Although no difference in cleavage rate, blastocyst formation rate and the average number of cells/blastocysts, the hatching blastocyst rate was different between the breeds (P > 0.05). In the Co BinhThuan breed, the rate of pregnancy of ejaculated groups was similar to that of the epididymal group. In conclusion, the ejaculated and epididymal sperm of native Vietnamese pigs were successfully frozen. We succeeded in creating embryos in vitro and pregnant pigs after artificial insemination from frozen-thawed semen in three native Vietnamese pig breeds for the first time. The use of the ejaculated sperm improved the production of native pig embryos in vitro efficiency.

Lipoprotein (a) Cut-Off in Chronic Kidney Disease Patients with a History of Cardiovascular Disease in Center Hospital University Souro SANOU, Burkina Faso

Patients living with chronic kidney disease (CKD) are at high risk of cardiovascular events. Our aim in this study was to assess the cut-off value for lipoprotein (a) (Lp(a)) in CKD patients with a history of cardiovascular disease (CVD). This was a cross-sectional study. Variables including age, sex, history of CVD, body mass index and CKD stage, were collected during CKD patient’s first admission in the nephrology dialysis department. Blood samples were collected for quantitative determination of Lp(a) by immunoturbidimetric method. They were divided into two groups: CKD patients without history of CVD and CKD patients with history of CVD. Fisher’s exact test was used to assess associations with a significance level of 0.05%. Area under the curve (AUC) and new cut-off value for Lp(a) were identified by drawing Receiver Operating Characteristic (ROC) curve. A total of seventy CKD patients with median age of 43 years [minimum-maximum = 15 - 78 years] were included. Patients with history of CVD were 65.71% (46/70). New Lp(a) cut-off point in CKD patients with history of CVD was 66.50 nmol/L [sensitivity, 87.00%;specificity, 58.30%;AUC = 0.727;p = 0.000]. ROC curve demonstrated good performance of Lp(a) to screen CKD patients with history of CVD. Further research is needed to determine an LPA gene polymorphism’s contribution to increasing risk for CVD at each kidney disease stage.

Impact of Four Fiber Sources and the Strategy of Feeding on the Nutritional Quality of Rabbit Meat (Oryctogalagus cuniculis)

This study was conducted to valorize four sources of fiber (Gliricidia sepium, Leucaena leucocephala, Moringa oleifera and palm nut fibers) in animal production. The experiments were carried out on 128 rabbits divided into 4 batches of 32 animals, reared for 8 weeks under the same conditions. Each batch was subdivided into two subgroups, one fed with a complete diet (a diet containing one of the fiber sources and served all day) and the other with the same diet separated from the fiber source (served at 9 a.m. and supplemented with the fiber source at 4 p.m.). Eight (8) experimental rations were, respectively, tested on the subgroups: complete feed Gliricidia sepium (CFG);supplemented feed Gliricidia sepium (SFG);complete feed Leucaena leucocephala (CFL);supplemented feed Leucaena leucocephala (SFL);complete feed Moringa oleifera (CFM);supplemented feed Moringa oleifera (SFM);complete feed palm nut fiber (CFF);supplemented feed palm nut fiber (SFF). In each subgroup, 4 rabbits were slaughtered at 15 weeks of age for a total of 32 rabbits. Physico-chemical parameters were evaluated on the feed and the meat. Data were analyzed using SAS 2013 software. Fiber content was similar (p > 0.05) for complete feeds. Fat content was high (p < 0.001) for the palm nuts fiber (27.34%) and the CFF feed (11.36%). Feeding rabbits with G. sepium leaves or palm nut fiber continuously increased the fat content of the meat in contrast to sequential feeding. Meat quality was also better when the fiber source was used in the feed of the rabbits in the evening.

传染性支气管炎病毒核衣壳蛋白基因变异及在大场杆菌中的表达(英文)

核衣壳蛋白 (N)是传染性支气管炎病毒的主要结构蛋白之一。以前的研究结果显示 IBV- ZJ971毒株以引起腺胃肿大为特征 ,IBV- X和 N毒株以引起肾炎为特征。为了探讨不同组织嗜性传染性支气管炎病毒 N基因的变异 ,我们依据已发表的 IBV- Beaudette毒株 N基因的序列设计和合成引物 ,RT-PCR扩增 IBV- ZJ971、N、X和 H5 2的核衣壳蛋白基因 ,并测序、分析和在 E.coli中进行表达。结果显示 :IBV- ZJ971、N、X和 H5 2毒株的 N基因的 ORF由 12 30 bp组成 ,编码 4 0 9个氨基酸。与来源于 Gen Bank中的 IBV毒株比较 ,IBV- ZJ971、N、X和 H5 2与其它 IBV毒株的核苷酸同源性分别是 87.0 - 98.6 %、86 .6 - 99.7%、86 .3- 99.7%、87.2 % - 98.5 % ,氨基酸的同源性分别是 90 .0 - 97.8%、 90 .5 - 98.8%、 90 .0 -98.8%、 91- 98%。IBV- ZJ971毒株的 N蛋白不同于其他 IBV的独特变异是 111,117,14 2 ,171和 4 0 1位点上的氨基酸改变 ,IBV- N和 X毒株的 N蛋白不同与其它 IBV的独特变异是 4 6 ,4 8,189,190 ,2 2 0 ,2 2 3,2 36 ,2 37,2 4 0 ,2 86 ,2 99,30 1334,335 ,336 and35 1位点上的氨基酸改变。说明 IBV嗜腺胃毒株(ZJ971)和嗜肾毒株 (N和 X)的 N基因以散在的点突变为特征。将 IBV- ZJ971毒株的 N基因在 E.

Genetic Variation of the VP1 Gene of the Virulent Duck Hepatitis A Virus Type 1(DHAV-1) Isolates in Shandong Province of China

To investigate the relationship of the variation of virulence and the external capsid proteins of the pandemic duck hepatitis A virus type 1(DHAV-1) isolates,the virulence,cross neutralization assays and the complete sequence of the virion protein 1(VP1) gene of nine virulent DHAV-1 strains,which were isolated from infected ducklings with clinical symptoms in Shandong province of China in 2007-2008,were tested.The fifth generation duck embryo allantoic liquids of the 9 isolates were tested on 12-day-old duck embryos and on 7-day-old ducklings for the median embryonal lethal doses(ELD 50 s) and the median lethal doses(LD 50 s),respectively.The results showed that the ELD 50 s of embryonic duck eggs of the 9 DHAV-1 isolates were between 1.9 × 10 6 /mL to 1.44 × 10 7 /mL,while the LD 50 s were 2.39 × 10 5 /mL to 6.15 × 10 6 /mL.Cross-neutralization tests revealed that the 9 DHAV-1 isolates were completely neutralized by the standard serum and the hyperimmune sera against the 9 DHAV-1 isolates,respectively.Compared with other virulent,moderate virulent,attenuated vaccine and mild strains,the VP1 genes of the 9 strains shared 89.8%-99.7% similarity at the nucleotide level and 92.4%-99.6% at amino acid level with other DHAV-1 strains.There were three hypervariable regions at the C-terminus(aa 158-160,180-193 and 205-219) and other variable points in VP1 protein,but which didn't cause virulence of DHAV-1 change.