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Proteomic identification of ruminal epithelial protein expression profiles in response to starter feed supplementation in pre-weaned lambs

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animal Nutrition 2021年第4期7卷 1271-1282页

作者： Daming Sun Shengyong Mao Weiyun Zhu Junhua Liu Jiangsu Key laboratory of Gastrointestinal Nutrition and animal Health Laboratory of Gastrointestinal MicrobiologyCollege of Animal Science and TechnologyNanjing Agricultural UniversityNanjing 210095Jiangsu ProvinceChina National Center for International Research on animal Gut Nutrition Nanjing Agricultural UniversityNanjing 210095China National Experimental Teaching Demonstration Center of animal Science Nanjing Agricultural UniversityNanjing 210095China

The pre sent study aimed to comparatively characterize the ruminal epithelial protein expression profiles in lambs fed ewe milk or milk plus starter diet using proteome *** new-born lambs were randomly divided into a group receiving ewe milk(M,n=10)and a group receiving milk plus starter diet(M+S,n=10).From 10 d old,M group lambs remained with the ewe and suckled ewe milk without receiving the starter *** lambs in the M+S group were separated from the ewe and received starter *** lambs were slaughtered at 56 d *** rumen epithelia samples(4 per group)were collected to characterize their protein expression profiles using proteomic *** analysis showed that 31 upregulated proteins and 40 downregulated proteins were identified in the rumen epithelium of lambs in response to starter diet *** results showed that starter feeding regulates a variety of biological processes in the epithelium,especially blood vessel development and extracellular matrix protein ***,the expression of proteins associated with synthesis and degradation of ketone bodies,butanoate metabolism,and citrate cycle signaling transduction pathway were upregulated in the group with starter diet supplementation,including 3-hydroxy-3-methylglutaryl coenzyme A synthase(HMGCS2,fold change[FC]=1.93),3-hydroxybutyrate dehydrogenase 1(BDH1,FC=1.91),and isocitrate dehydrogenase 1(IDH1,FC=8.12).The metabolic processes associated with ammonia detoxification and antioxidant stress were also affected by starter diet supplementation,with proteins,microsomal glutathione S-transferase 3(MGST3,FC=2.37)and IDH1,linked to the biosynthesis of glutamate and glutathione metabolism pathway being upregulated in the group with starter diet *** addition,starter feeding decreased the expression of Rasrelated protein rap-1 A(RAP1 A,FC=0.48)enriched in Rap1 signaling pathway,Ras signaling pathway,cyclic adenosine monophosphate(cAMP)signaling pa

关键词： Starter Rumen epithelium Protein expression Proteomics Lamb

Effects of high‑grain diet feeding on fatty acid profiles in milk,blood,muscle,and adipose tissue,and transcriptional expression of lipid‑related genes in muscle and adipose tissue of dairy cows

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Journal of animal Science and Biotechnology 2023年第5期14卷 2063-2078页

作者： Qiaorong Cui Limei Lin Zheng Lai Shengyong Mao Jiangsu Key laboratory of Gastrointestinal Nutrition and animal Health Laboratory of Gastrointestinal MicrobiologyCollege of Animal Science and TechnologyNanjing Agricultural UniversityNanjingJiangsu Province 210095China National Center for International Research on animal Gut Nutrition Nanjing Agricultural UniversityNanjing 210095China National Experimental Teaching Demonstration Center of animal Science Nanjing Agricultural UniversityNanjing 210095China

Background High-grain(HG)diets affect lipid metabolism in the liver and mammary tissue of dairy cows,but its effects on muscle and adipose tissue have not been wide ***,the aim of this study is to clarify this *** Twelve Holstein cows were randomly divided into two groups:conventional diet group(CON,n=6)and the HG diet group(n=6).On day 7 of week 4,rumen fluid was sampled to measure pH,milk was sampled to meas-ure components,and blood was sampled to measure biochemical parameters and fatty acid *** the experiment,cows were slaughtered to collect muscle and adipose tissue for fatty acid composition and transcriptome *** HG feeding decreased the ruminal pH,milk’s fat content and long-chain fatty acid proportion(P<0.05)and increased the proportion of short-and medium-chain fatty acids in the milk(P<0.05)as compared with CON *** concentrations of blood cholesterol,low-density lipoprotein,and polyunsaturated fatty acids in the HG cows were lower than those in CON cows(P<0.05).In muscle tissue,HG feeding tended to increase the triacylglycerol(TG)concentration(P<0.10).Transcriptome analysis revealed changes in the biosynthesis of the unsaturated fatty acids pathway,the regulation of lipolysis in the adipocytes pathway,and the PPAR signalling *** adipose tissue,HG feeding increased the concentration of TG and decreased the concentration of C18:1 cis9(P<0.05).At the transcrip-tome level,the fatty acid biosynthesis pathway,linoleic acid metabolism pathway,and PPAR signalling pathway were *** HG feeding leads to subacute rumen acidosis and a decreased milk fat *** fatty acid profiles in the milk and plasma of dairy cows were changed by HG *** muscle and adipose tissue,HG feeding increased TG concentration and up-regulated the expression of genes related to adipogenesis,while down-regulated the expression of genes related to lipid *** results complement our knowledge of the

关键词： Adipose tissue Fatty acid composition High-grain diets Lipid transcriptional profiles Muscle tissue

Construction and Characterization of a Novel Bacmid AcBac-Syn Based on a Synthesized Baculovirus Genome

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Virologica Sinica 2021年第6期36卷 1566-1574页

作者： Yu Shang Hengrui Hu Xi Wang Hualin Wang Fei Deng Manli Wang Zhihong Hu Key laboratory of Prevention and Control Agents for animal Bacteriosis(Ministry of Agriculture and Rural Affairs)&Hubei Provincial Key laboratory of animal Pathogenic microbiology Institute of Animal Husbandry and VeterinaryHubei Academy of Agricultural SciencesWuhan 430064China State Key laboratory of Virology Wuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China University of Chinese Academy of Sciences Beijing 100049China

Baculoviruses are large DNA viruses which have been widely used as expression vectors and biological *** recombination and Bac-to-Bac system have been the main methods for manipulating the baculovirus ***, we generated a synthetic baculovirus Ac MNPV-WIV-Syn1 which fully resembled its parental virus Autographa californica multiple nucleopolyhedrovirus(Ac MNPV). Here, we report the modification of Ac MNPV-WIV-Syn1 into a novel bacmid, Ac Bac-Syn, which can be used as a backbone for Bac-to-Bac system. To achieve this, a vector contained a Lac Z:att Tn7 and egfp cassette was constructed, and recombined with a linearized Ac MNPV-WIV-Syn1 genome by transformation-associated recombination in yeast to generate bacmid Ac Bac-Syn. The bacmid was then transfected to insect cells and the rescued virus showed similar biological characteristics to the wild-type virus in terms of the kinetics of budded virus production, the morphology of occlusion bodies, and the oral infectivity in insect larvae. For demonstration, a red fluorescent protein gene Dsred was transposed into the att Tn7 site by conventional Bac-to-Bac method, and the transfection and infection assays showed that Ac Bac-Syn can be readily used for foreign gene insertion and *** Bac-Syn has several advantages over the conventional Ac MNPV bacmids, such as it contains an egfp reporter gene which facilitates visualization of virus propagation and titration;its DNA copy numbers could be induced to a higher level in E. coli;and the retaining of the native polyhedrin gene in the genome making it an attractive system for studying the functions of gene related to occlusion body assembly and oral infection.

关键词： Synthetic genome Baculovirus Autographa californica multiple nucleopolyhedrovirus(AcMNPV) Large DNA virus

The temporal dynamics of antimicrobial-resistant Salmonella enterica and predominant serovars in China

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National Science Review 2023年第3期10卷 60-76页

作者： Yanan Wang Yue Liu Na Lyu Zhiyuan Li Sufang Ma Demin Cao Yuanlong Pan Yongfei Hu Hua Huang George F.Gao Xuebin Xu the Bacterium-learning Union Baoli Zhu International Joint Research Center of National animal Immunology College of Veterinary MedicineHenan Agricultural University CAS Key laboratory of Pathogen microbiology and Immunology Institute of Microbiology Chinese Academy of Sciences Department of microbiology Shanghai Municipal Center for Disease Control and Prevention Savaid Medical School University of Chinese Academy of Sciences State Key laboratory of animal Nutrition College of Animal Science and Technology China Agricultural University Beijing Products Quality Supervision and Inspection Institute Chinese Center for Disease Control and Prevention(China CDC) Beijing Key laboratory of Antimicrobial Resistance and Pathogen Genomics Department of Pathogenic Biology School of Basic Medical Sciences Southwest Medical University

Salmonella enterica is one of the most common bacterial pathogens in humans and *** studies on the trends and geographical distribution of antimicrobial-resistant Salmonella and dominant serovars have been well studied in European and American countries while not in ***,taking the One-Health strategy,we used>35 000 Salmonella enterica isolates to explore the temporal and spatial dynamics of dominant serovars in *** found that Salmonella Typhimurium was the dominant serovar causing human infection in China,which was consistent with Australia but inconsistent with North American and European *** proportion of Salmonella serovars Typhimurium,London,Rissen,Corvallis,Meleagridis,Kentucky,and Goldcoast showed an increasing trend during *** randomly selected 1962 isolates for comparative genomics and antimicrobial resistance studies and found that the number of antibiotic resistance genes(ARGs) per isolate increased 1.84 and 2.69 times of human and non-human origins,respectively,spanning 14 *** proportion of antimicrobial-resistant Salmonella isolates had an increasing trend during 2006-2019,especially beta-lactam,quinolone,tetracycline,and rifampicin ***,we found that higher diversity of sequence types(STs) in *** than in other serovars,ST34 from pig and ST 19 from chicken origin,were mainly associated with isolates causing child and adult gastro-infection,*** results fill in the data gap on the trends of dominant serovars and antimicrobial resistance of Salmonella enterica in *** data provide useful information for public health decision-makers prioritizing interventions for foodborne diseases and food safety.

关键词： salmonella enterica food safety antimicrobial resistance whole-genome sequencing public health

Effects of dietary replacement of soybean meal with dried distillers grains with solubles on the microbiota occupying different ecological niches in the rumen of growing Hu lambs

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Journal of animal Science and Biotechnology 2021年第2期12卷 677-688页

作者： Junshi Shen Zhipeng Li Zhongtang Yu Weiyun Zhu laboratory of Gastrointestinal microbiology Jiangsu Key Laboratory of Gastrointestinal Nutrition and Animal HealthCollege of Animal Science and TechnologyNanjing Agricultural UniversityNanjing 210095China National Center for International Research on animal Gut Nutrition Nanjing Agricultural UniversityNanjing 210095China Department of Special animal Nutrition and Feed Science Institute of Special Animal and Plant SciencesChinese Academy of Agricultural SciencesChangchun 130112China Department of animal Sciences The Ohio State UniversityColumbusOH 43210USA

Background:Diet has a profound impact on the rumen microbiota,and the impact can vary among the different rumen ecological niches(REN).This study investigated the effects of dietary replacement of soybean meal(SBM)with dried distillers grains with solubles(DDGS)on the rumen microbiota occupying different REN of growing Hu *** a 9-week feeding trial,6 lambs from each dietary treatment(SBM ***-based diets)were slaughtered for sample *** microbiota of the rumen solid,liquid,and epithelium fractions was examined using amplicon sequencing analysis of bacterial 16S rRNA gene,functional prediction,and ***:No interaction of dietary protein source(PS)and REN were detected for virtually all the measurements made in this *** DDGS substitution resulted in very limited influence on bacterial community ***,the metabolic pathways predicted from 16S rRNA gene sequences varied greatly between SBM-and DDGS-based *** populations of rumen total bacteria,fungi,sulfate-reducing bacteria(SRB),and methanogens were not influenced by DDGS substitution,but the population of protozoa was *** bacterial communities in rumen solid(RS)and liquid(RL)were similar in taxonomic composition but were different in relative abundance of some *** contrast,the bacterial composition and relative abundance of rumen epithelium(RE)were greatly distinct from those of the RS and the *** alignment with the bacterial relative abundance,the metabolic pathways predicted from 16S rRNA genes also varied greatly among the different *** populations of total bacteria,protozoa,and methanogens attached to the RE were smaller than those in the RS and RL,and the fungal population on the rumen epitheliumwas smaller than that in the RS but similar to that in the *** the contrary,the SRB population on the RE was greater than that in the RS and ***:Substitution of SBM with DDGS had greater impact to the protozoa than to the other microbes,and

关键词： Distillers dried grains with solubles Growing lamb Microbiota Ruminal ecological niche Soybean meal

Screening and characterization of a novel ruminal cellulase gene(Umcel-1) from a metagenomic library of gayal(Bos frontalis)

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Journal of Integrative Agriculture 2016年第4期15卷 855-861页

作者： LI Bi-feng ZHU Ya-xin GU Zhao-bing CHEN Yuan LENG Jing GOU Xiao FENG Li LI Qing XI Dong-mei MAO Hua-ming YANG Shu-Li Yunnan Provincial Key laboratory of animal Nutrition and Feed Science Yunnan Agricultural University Institute of microbiology Chinese Academy of Sciences Department of Biological Science Huizhou University

Gayal is a rare semi-wild bovine species found in the Indo-China. They can graze grasses, including bamboo leaves, as well as reeds and other plant species, and grow to higher mature live weights than Yunnan Yellow cattle maintained in similar harsh environments. The aim of this study was to identify specific cellulase in the gayal rumen. A metagenomic fosmid library was constructed using genomic DNA isolated from the ruminal contents of four adult gayals. This library contained 38400 clones with an average insert size of 35.5 kb. The Umcel-1 gene was isolated from this library. Investigation of the cellulase activity of 24 random clones led to the identification of the Umcel-1 gene, which exhibited the most potent cellulase activity. Sequencing the Umcel-1 gene revealed that it contained an open reading frame of 942 base pairs that encoded a product of 313 amino acids. The putative gene Umcel-1 product belonged to the glycosyl hydrolase family 5 and showed the highest homology to the cellulase （GenBank accession no. YP_004310852.1 ） from Clostridium lentocellum DSM 5427, with 44% identity and 62% similarity. The Umcel-1 gene was heterologously expressed in Escherichia coil BL21, and recombinant Umcel-1 was purified. The activity of purified recombinant Umcel-1 was assessed, and the results revealed that it hydrolyzed carboxymethyl cellulose with optimal activity at pH 5.5 and 45~C. To our knowledge, this study provides the first evidence for a cellulase produced by bacteria in gayal rumen.

关键词： gayal ruminal microorganism fosmid library Umcel-I gene gene expression cellulase activity

Understanding the metabolism of Mycoplasma mycoides subsp. capri in vitro by a transcriptomic analysis

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Journal of Integrative Agriculture 2018年第2期17卷 428-435页

作者： WANG Xiao-hui WANG Yan-fang HUANG Hai-bi BAI Fan SHI Xiao-na MA Chang-jiao GAO Yuan ZHANG Jian-hua ZHANG Wen-guang HAO Yong-qing Key laboratory of microbiology & Immunology College of Veterinary Medicine Inner Mongolia Agricultural University Foundation Medicine College Baotou Medical School Key laboratory of animal Genetics and Genomics Inner Mongolia Agricultural University

It is generally known that the culture for mycoplasma is time-consuming and a variety of nutrients are needed in the culture medium. This brings a lot of difficulties to mycoplasma research and application, including Mycoplasma mycoides subsp. capri（Mmc）. Furthermore, little research on the characteristics of Mmc metabolism has been reported. In this study, Mmc PG3 strain cultures were investigated for dynamic gene expression. Culture samples were harvested during logarithmic phase（PG3-1）, stationary phase（PG3-2）, decline phase（PG3-3） and late decline phase（PG3-4）. Twelve RNA samples（three replicates for each of the four growth stages considered） from these cultures were collected and sequenced. Paired comparison between consecutive growth phases in the four growth stages showed 45 significant differentially expressed genes（P〈0.01） were linked to PG3 metabolism. The enzymes these genes coded were mainly involved in ATP synthase, pyrimidine metabolism, nicotinate and nicotinamide metabolism, arginine and proline metabolism. Among these, cytidylate kinase, fructose 1,6-bisphosphate aldolases Class II, nicotinate-nucleotide adenylyltransferase and dihydrolipoamide dehydrogenase play a key role in Mmc metabolism. These results provide a baseline to build our understanding of the metabolic pathway of Mmc.

关键词： metabolism Mycoplasma mycoides subsp.capri transcriptomic

Mutagenic and cytotoxic potential of Endosulfan and Lambda-cyhalothrin—In vitro study describing individual and combined effects of pesticides

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Journal of Environmental Sciences 2014年第7期26卷 1471-1479页

作者： Umber Saleem Sohail Ejaz Muhammad Ashraf Muhammad Ovais Omer Imran Iltaf Zainab Batool Riffat Fatima Msbah Afzal Department of Pharmacology & Toxicology University of Veterinary and Animal Sciences Lahore 54000 Pakistan Department of Clinical Neurosciences Neurology Unit Addenbrooke's Hospital University of Cambridge Cambridge UK microbiology Section Quality Operations Laboratory University of Veterinary and Animal Sciences Lahore Pakistan

Excessive use of pesticides poses increased risks to non target species including humans. In the developing countries, lack of proper awareness about the toxic potential of pesticides makes the farmer more vulnerable to pesticide linked toxicities, which could lead to diverse pathological conditions. The toxic potential of a pesticide could be determined by their ability to induce genetic mutations and cytotoxicity. Hence, determination of genetic mutation and cytotoxicity of each pesticide is unavoidable to legislate health and safety appraisal about pesticides. The objective of current investigation was to determine the genotoxic and cytotoxic potential of Endosulfan（EN） and Lambda-cyhalothrin（LC）; individually and in combination. 3-（4,5-Dimethylthiazol-2-yl）-2,5-Diphenyltetrazolium Bromide（MTT） assay was utilized to determine cytotoxicity, while two mutant histidine dependent Salmonella strains（TA98, TA100） were used to determine the mutagenicity of EN and ***, mutagenicity assay was conducted with and without S9 to evaluate the effects of metabolic activation on mutagenicity. Even though a dose dependent increase in the number of revertant colonies was detected with EN against both bacterial strains, a highly significant（p 〈 0.05） increase in the mutagenicity was detected in TA98 with S9. In comparison, data obtained from LC revealed less mutagenic potential than EN. Surprisingly,the non-mutagenic individual-concentrations of EN and LC showed dose dependent mutagenicity when combined. Combination of EN and LC synergistically induced mutagenicity both in TA98 and TA100. MTT assay spotlighted comparable dose dependent cytotoxicity effects of both pesticides. Interestingly, the combination of EN and LC produced increased reversion and cytotoxicity at lower doses as compared to each pesticide, concluding that pesticide exposure even at sub-lethal doses can produce cytotoxicity and genetic mutations, which could lead to carcinogenicity.

关键词： Endosulfan Lambda-cyhalothrin Mutagenicity Cytotoxicity

CypA Regulates AIP4-Mediated M1 Ubiquitination of Influenza A Virus

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Virologica Sinica 2018年第5期33卷 440-448页

作者： Madina Mahesutihan Weinan Zheng Liang Cui Yun Li Pengtao Jiao Wenxian Yang Wei Liu Jing Li Wenhui Fan Limin Yang Wenjun Liu Lei Sun CAS Key laboratory of Pathogenic microbiology and Immunology Institute of MicrobiologyChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China State Key laboratory for Conservation and Utilization of Subtropical Agro-Bioresources and laboratory of animal Infectious Diseases College of Animal Sciences and Veterinary MedicineGuangxi UniversityNanning 530004China College of Life Sciences Henan Agricultural UniversityZhengzhou 450002China

Cyclophilin A (CypA) is a peptidyl-prolyl cis/trans isomerase that interacts with the matrix protein (M1) of influenza A virus (IAV) and restricts virus replication by regulating the ubiquitin–proteasome-mediated degradation of M1. However,the mechanism by which CypA regulates M1 ubiquitination remains unknown. In this study, we reported that E3 ubiquitin ligase AIP4 promoted K48-linked ubiquitination of M1 at K102 and K104, and accelerated ubiquitin–proteasome-mediated degradation of M1. The recombinant IAV with mutant M1 (K102 R/K104 R) could not be rescued, suggesting that the ubiquitination of M1 at K102/K104 was essential for IAV replication. Furthermore, CypA inhibited AIP4-mediated M1 ubiquitination by impairing the interaction between AIP4 and M1. More importantly, both the mutations of M1 (K102 R/K104 R) and CypA inhibited the nuclear export of M1, indicating that CypA regulates the cellular localization of M1 via inhibition of AIP4-mediated M1 ubiquitination at K102 and K104, which results in the reduced replication of ***, our findings reveal a novel ubiquitination-based mechanism by which CypA regulates the replication of IAV.

关键词： Influenza A virus (IAV) Ubiquitination Cyclophilin A (CypA) AIP4 M1