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Listeria monocytogenes Mutants Carrying Newcastle Disease Virus F Gene Fused to its actA and plcB:In vitro Expression and Immunogenicity in Chickens

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Acta Biochimica et Biophysica Sinica 2007年第1期39卷 57-66页

作者： Lingli JIANG Chunlin KE Jingjing XU Jianshun CHEN Xueyan CHEN Ning CHEN Jiangbing SHUAI and Weihuan FANG Institute of Preventive veterinary medicine,and Zhejiang Provincial Key Laboratory for Preventve veterinary medicine,Zhejiang university,Hangzhou 310029,China Institute of Preventive veterinary medicine and Zhejiang Provincial Key Laboratory for Preventive Veterinary Medicine Zhejiang UniversityHangzhou 310029 China

Recombinant Listeria monocytogenes mutants carrying Newcastle disease virus (NDV) fusionprotein gene F were constructed by homologous *** F or its truncated fragment Fa wasused as the model heterologous gene to be integrated into actA or plcB downstream of their signal *** orientation of the inserted genes was verified by polymerase chain reaction amplification of F or *** inserted F and Fa were expressed in the two recombinants Lm-AactA-F and Lm-AplcB-Fa as shown bysodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western *** recombinantsexhibited reduced virulence to embryonated eggs and mice by about 1.5-2.5 logs as compared with theparent wild strain 10403S. They were also less invasive than strain 10403S (P<0.05). Chickens receiving therecombinant strains orally or intraperitoneally were partially protected from virulent NDV challenge possiblydue to enhancement of non-specific immunity because the antibody titers against the homologous virus strainor the recombinant truncated fusion protein were marginal. Further research is needed in other animal modelsto see if the low antibody response results from insufficient expression of the heterologous genes as a resultof failure of L. monocytogenes or its recombinants to persist or replicate in chickens.

关键词： Listeria monocytogenes homologous recofflbination Newcastle disease virvs fusion protein F heterologous gene expression immunogenicity

Virulence Phenotyping and Molecular Characterization of a Low-pathogenicity Isolate of Listeria monocytogenes from Cow's Milk

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Acta Biochimica et Biophysica Sinica 2006年第4期38卷 262-270页

作者： Ling-Li JIANG Jing-Jing XU Ning CHEN Jiang-Bing SHUAI Wei-Huan FANG* Institute of Preventive veterinary medicine,Zhejiang Provincial Key Laboratory of Preventive veterinary medicine,Zhejiang university,Hangzhou 310029,China Institute of Preventive veterinary medicine Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine Zhejiang UniversityHangzhou 310029 China

A low-pathogenicity isolate of Listeria monocytogenes from cow’s milk,as screened in mouseand chicken embryonated egg models,was examined for virulence-related phenotypic *** genes (iap,prfA,plcA,hly,mpl,actA,plcB,InlA and InlB) were compared with *** strains 10403S and EGD to elucidate the possible molecular mechanisms of low ***-though *** H4 exhibited similar patterns to strain 10403S in terms of hemolytic activity,invitro growth and invasiveness and even had higher adhesiveness,faster intracellular growth and higherphospholipase activity in vitro,it was substantially less virulent than the strain 10403S in mouse and chickenembryo models (50% lethal dose:10vs.10and 10vs.10,respectively).The genes prfA,plcA andmpl were homologous among *** strains H4,10403S and EGD (>98%).Genes iap,hly,plcB,InlA and InIB of *** 10403S had higher homology to those of strain EGD (>98%) than *** homology of the gene hly between strain 10403S and isolate H4 was 96.9% at the nucleotide level,but 98.7% at the amino acid *** actA gene of isolate H4 had deletions of 105 nucleotides correspondingto 35 amino acid deletions falling Within the proline-rich *** together,this study presents someclues as to reduced virulence to mice and chicken embryos of the isolate H4 probably as a result of deletionmutations of actA.

关键词： Listeria monocytogenes virulence factor virulence-associated gene

Aldehyde dehydrogenase activity helps identify a subpopulation of murine adipose-derived stem cells with enhanced adipogenic and osteogenic differentiation potential

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World Journal of Stem Cells 2017年第10期9卷 179-186页

作者： Harumichi Itoh Shimpei Nishikawa Tomoya Haraguchi Yu Arikawa Shotaro Eto Masato Hiyama Toshie Iseri Yoshiki Itoh Munekazu Nakaichi Yusuke Sakai Kenji Tani Yasuho Taura Kazuhito Itamoto Department of Small Animal Clinical Science Joint Faculty of Veterinary Medicine Yamaguchi University Department of veterinary Surgery Joint Faculty of Veterinary Medicine Yamaguchi University Department of veterinary Radiology Joint Faculty of Veterinary Medicine Yamaguchi University Department of veterinary Pathology Joint Faculty of Veterinary Medicine Yamaguchi University

AIMTo identify and characterize functionally distinct subpopulation of adipose-derived stem cells (ADSCs). METHODSADSCs cultured from mouse subcutaneous adipose tissue were sorted fluorescence-activated cell sorter based on aldehyde dehydrogenase (ALDH) activity, a widely used stem cell marker. Differentiation potentials were analyzed by utilizing immunocytofluorescece and its quantitative analysis. RESULTSApproximately 15% of bulk ADSCs showed high ALDH activity in flow cytometric analysis. Although significant difference was not seen in proliferation capacity, the adipogenic and osteogenic differentiation capacity was higher in ALDHHi subpopulations than in ALDHLo. Gene set enrichment analysis revealed that ribosome-related gene sets were enriched in the ALDHHi subpopulation. CONCLUSIONHigh ALDH activity is a useful marker for identifying functionally different subpopulations in murine ADSCs. Additionally, we suggested the importance of ribosome for differentiation of ADSCs by gene set enrichment analysis.

关键词： Adipose-derived stem/stromal cell Aldehyde dehydrogenase activity Flow cytometry Subpopulation Ribosome

Clinicobiochemical investigations of gangrenous mastitis in does:immunological responses and oxidative stress biomarkers

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Journal of Zhejiang university-Science B(Biomedicine & Biotechnology) 2013年第1期14卷 33-39页

作者： Wael M.EL-DEEB Department of Clinical Studies College of Veterinary Medicine and Animal ResourcesKing Faisal University Department of veterinary medicine Infectious Diseases and Fish DiseasesFaculty of Veterinary MedicineMansoura University

A total of 50 does were used to determine selected hematological and biochemical parameters with special references to oxidative stress markers, acute phase protein profiles, and proinflammatory cytokines in healthy and gangrenous mastitis affected does. Animals were divided into two equal groups represented as clinically healthy （control） and diseased groups, respectively. The bacteriological examination of milk samples from diseased does revealed many types of bacterial infection. The isolated bacteria were Staphy/ococcus aureus （N=23/25）, Escherichia coil （N=11/25）, and Clostridium perfringens （N=4/25）. There was a significant increase in the levels of β-hydrexybutyrate, non-esterified free fatty acids, triglyceride, low density lipoprotein cholesterol （LDL-C）, aspartate aminotransferase, and alanine aminotransferase and a significant reduction in the levels of glucose, cholesterol, and high density lipoprotein cholesterol （HDL-C） in does with gangrenous mastitis compared to healthy does. Moreover, there was a significant increase in the levels of malondialdehyde and uric acid with a significant decrease in the levels of reduced glutathione, super oxide dismutase, and catalase in does with gangrenous mastitis compared to healthy does. In addition, there was a significant increase in the haptoglobin, serum amyloid A, fibrinogen, interleukin 6 （IL-6）, IL-1β, and tumor necrosis factor-a （TNF-a） in does with gangrenous mastitis compared to healthy ones. Conclusively, oxidative stress biomarkers, acute phase proteins, and proinflammatory cytokines play an essential task as biomarkers for gangrenous mastitis in does. Mastitis may be considered as one of the ketotic stressors in does after parturition.

关键词： Haptoglobin Serum amyloid A Malondialdehyde Interleukin 1β Interleukin 6 Tumor necrosis factor-a

Pharmacokinetics and Residues of Cefquinome in Milk of Lactating Chinese Dairy Cows After Intramammary Administration

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Journal of Integrative Agriculture 2014年第12期13卷 2750-2757页

作者： LI Ya-fei WANG Lin GU Xiao-yan ZENG Zhen-ling HE Li-min YANG Fan YUAN Bo SHU Jian-hua DING Huan-zhong Laboratory of veterinary Pharmacology College of Veterinary Medicine South China Agricultural University

The purpose of the study was to investigate the pharmacokinetics of cefquinome in plasma and milk samples of lactating Chinese Holstein following a single intramammary administration into one quarter at the dose of 75 mg. Residue depletion of cefquinome in milk administrated at one quarter following three consecutive infusions at the same dose were also carried out. Cefquinome concentrations in plasma and milk were determined by high-performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） method. A non-compartmental analysis was used to obtain the pharmacokinetic parameters of cefquinome. Following the single treatment, cefquinome wasn’t detected in any of the plasma samples. The concentration of cefquinome in milk reached peaked values （Cmax） of （599.00±322.00） μg mL-1 at 2 h after administration （Tmax）, elimination half-life （t1/2λz） was （4.63±0.26） h, area under the concentration-time curve （AUC0-∞） was （4 890.19±1 906.98） μg mL-1 h, and mean residence time （MRT） was （6.03±2.27） h. In residue depletion study, cefquinome concentrations in 5 out of 6 milk samples at 72 h were lower than the maximum residue limit ifxed by the European regulatory agency （20μg kg-1 for cefquinome） and cefquinome still could be detected in milk of treated quarters at 120 h post-treatment. The maximum concentration （Cmax） of cefquinome in milk from treated quarters was （486.50±262.92） μg mL-1 and arrived at 6 h after administration （Tmax）, elimination half-life （t1/2λz） was （6.30±0.76） h, area under the concentration-time curve （AUC0-∞） was （44 747.79±11 434.43） μg mL-1 h, and mean residence time （MRT） was （10.09±1.40） h. This study showed that cefquinome has the feature of poor penetration into blood and was eliminated quickly from milk in lactating cows after intramammary administration.

关键词： cefquinome pharmacokinetics residue dairy cows HPLC-MS/MS

Changing dietary n-6:n-3 ratio using different oil sources affects performance,behavior,cytokines mRNA expression and meat fatty acid profile of broiler chickens

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Animal Nutrition 2018年第1期4卷 -页

作者： Doaa Ibrahim Rania El-Sayed Safaa I.Khater Enas N.Said Shefaa A.M.El-Mandrawy Department of Nutrition and Clinical Nutrition Faculty of Veterinary MedicineZagazig University Department of Biochemistry Faculty of Veterinary MedicineZagazig University Department of veterinary Public Health Faculty of Veterinary MedicineZagazig University Department of Clinical Pathology Faculty of Veterinary MedicineZagazig University

Typical formulated broiler diets are deficient in n-3 poly-unsaturated fatty acids(PUFA) due to widening n-6:n-3 PUFA ratio which could greatly affect performance,immune system of birds and,more importantly,meat *** study was conducted to evaluate the effect of modifying dietary n-6:n-3 PUFA ratio from plant and animal oil sources on performance,behavior,cytokine m RNA expression,antioxidative status and meat fatty acid profile of broiler ***(n=420) were fed 7 diets enriched with different dietary oil sources and ratios as follows: sunflower oil in control diet(C); fish oil(FO); 1:1 ratio of sunflower oil to FO(C1 FO1); 3:1 ratio of sunflower oil to fish oil(C3 FO1); linseed oil(LO); 1:1 ratio of sunflower oil to linseed oil(C1 LO1); 3:1 ratio of sunflower oil to linseed oil(C3 LO1),resulting in dietary n-6:n-3 ratios of approximately 40:1,1.5:1,4:1,8:1,1:1,2.5:1 and 5:1,*** best final body weight,feed conversion ratio as well as protein efficiency ratio of broilers were recorded in the C1 FO1 and C1 LO1 *** with the control group,the dressing percentage and breast and thigh yield were highest in the C1 FO1 and C1 LO1 *** the dietary n-6:n-3 ratio increased(P < 0.05) n-3 PUFA content of breast ***,the breast meat contents of eicosapentaenoic acid and docosahexaenoic acid increased(P < 0.05) with increasing dietary FO whereas a-linolenic acid content was higher with LO ***,enriching the diets with n-3 PUFA from FO and LO clearly decreased(P < 0.05) serum total cholesterol,triglycerides and very low-density lipoproteins and enhanced antioxidative *** feeding frequency was decreased(P < 0.05) in the C1 FO1 and C1 LO1 ***,n-3 PUFA-enriched diets enhanced the frequency of preening,wing flapping and *** oil source addition,compared to plant oil,to broiler diets enhanced the relative m RNA expression of interferon gamma,interleukin-1 beta,interleukin-2 and

关键词： Broiler n-6:n-3 PUFA ratio Performance Antioxidant status Meat Immunity

Optimization of culture conditions for short-term maintenance, proliferation, and colony formation of porcine gonocytes

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Journal of Animal Science and Biotechnology 2018年第2期9卷 326-338页

作者： Awang Hazmi Awang-Junaidi Ali Honaramooz Department of veterinary Biomedical Sciences Western College of Veterinary Medicine University of Saskatchewan

Background: Gonocytes give rise to spermatogonial stem cells, and thereby play an essential role in establishing spermatogenesis. Optimized culture conditions for gonocytes provide an opportunity for their study and in vitro manipulation for potential application in reproductive technologies. Using six experiments in a step-wise design, we examined the effects of several culture conditions on the maintenance, proliferation, and colony formation of porcine gonocytes. Testis cells from neonatal piglets were cultured for 7 d in DMEM supplemented with 10% fetal bovine serum. The examined culture conditions included using different cell seeding densities, gonocyte proportions, incubation temperatures, sampling strategies, and medium changing ***: Confluency of cel s was optimal(>90% by ~6 d) when 3.0 × 104 testis cel s/cm2 containing ~40% gonocytes were used. Incubating the cel s at 35 °C or 37 °C resulted in similar cel number and viability at confluency, but incubation at 35 °C resulted in a delayed confluency. In the first 2 d of culture, gonocytes remained mostly floating in the medium and gradual y settled over the next 5 d. Consequently, not changing the medium for 7 d(as opposed to changing it every 2 d) led to a significant increase in the number of gonocyte colonies by reducing the loss of "floating gonocytes".Conclusion: We found that gonocytes require the presence of a critical minimum number of somatic cel s for settlement, and can proliferate and form growing colonies even in a basic medium. Large numbers of viable gonocytes remain floating in the medium for several days. The optimized culture conditions in the present study included seeding with 3.0 × 104 testis cel s/cm2 containing ~40% gonocytes, incubating at 37 °C, and without changing the medium in the first week, which can result in improved colony formation of porcine gonocytes.

关键词： Cell culture Male Germline stem cells Pigs Testis

Winery by‑products as a feed source with functional properties:dose–response effect of grape pomace,grape seed meal,and grape seed extract on rumen microbial community and their fermentation activity in RUSITEC

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Journal of Animal Science and Biotechnology 2023年第6期14卷 2439-2453页

作者： Ratchaneewan Khiaosa‑ard Mubarik Mahmood Elsayed Mickdam Catia Pacifico Julia Meixner Laura‑Sophie Traintinger Institute of Animal Nutrition and Functional Plant Compounds Department for Farm Animals and Veterinary Public HealthUniversity of Veterinary Medicine ViennaVeterinaerplatz 11210 ViennaAustria. Animal Nutrition Section Department of Animal SciencesSub Campus JhangUniversity of Veterinary and Animal Sciences12 Km Chiniot RoadJhang 35200LahorePakistan. Nutrition and Clinical Nutrition Department Faculty of Veterinary MedicineSouth Valley UniversityQena 83523Egypt. Present address:Biome Diagnostics GmbH ViennaAustria.

Background Grape and winery by-products have nutritional values for cattle and also contain functional compounds like phenols,which not only bind to protein but can also directly affect microbiota and their function in the *** characterized the nutritional and functional effects of grape seed meal and grape pomace as well as an effective dosage of grape phenols on ruminal microbiota and fermentation characteristics using a rumen simulation *** Six diets(each n=8)were compared including a control diet(CON,no by-product),a positive control diet(EXT,CON+3.7%grape seed extract on a dry matter(DM)basis),two diets with grape seed meal at 5%(GS-low)and 10%(GS-high),and two diets with grape pomace:at 10%(GP-low)and 20%(GP-high),on a DM *** inclusion of the by-product supplied total phenols at 3.4%,0.7%,1.4%,1.3%,and 2.7%of diet DM for EXT,GS-low,GS-high,GP-low,and GP-high,*** were tested in four experimental *** treatments decreased ammonia concentrations and the disappearances of DM and OM(P<0.05)compared to *** and GP-high lowered butyrate and odd-and branch-chain short-chain fatty acids while increased acetate compared to CON(P<0.05).Treatments did not affect methane *** decreased the abundance of many bacterial genera including those belonging to the core ***-high and EXT consistently decreased Olsenella and Anaerotipes while increased Ruminobacter *** The data suggest that the inclusion of winery by-products or grape seed extract could be an option for reducing excessive ammonia *** to grape phenols at a high dosage in an extract form can alter the rumen microbial ***,however,does not necessarily alter the effect of grape phenols on the microbial community function compared to feeding high levels of winery *** suggests the dominant role of dosage over the form or source of the grape phenols in affecting ruminal microbial *** conclusion,

关键词： Functional feed Grape pomace Grape seed Protein degradation Rumen microbiota

Development of a Rapid Multi-residue Assay for Detecting β-lactams Using Penicillin Binding Protein 2x

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Biomedical and Environmental Sciences 2013年第2期26卷 100-109页

作者： ZENG Kum ZHANG Jing WANG Yang WANG ZhanHui ZHANG Su Xia WU Chong Ming SHEN Jian Zhong Department of veterinary Pharmacology and Toxicology College of Veterinary MedicineChina Agricultural University

Objective To develop a rapid multi-residue assay for detecting 16 demanded by the European Union （EU）. Methods A recombinant penicillin-binding protein （PBP） 2x＊ from Streptococcus pneumoniae R6 was expressed in vitro and six β-1actams were conjugated to HRP by four methods. A rapid multi-residue assay for β-1actams was established with PBP2x＊ and HRP-conjugate. Results PBP2x＊ was expressed and purified successfully and the ideal HRP-conjugate was identified. The multi-residue assay was developed. After optimization, penicillin G, ampicillin, amoxicillin, cloxacillin, dicloxacillin, oxacillin, nafcillin, cephalexin, ceftiofur, cefalonium, cefquinome, cefazolin, cefoperazone, cephacetrile, and cephapirin can be detected at levels below MRL in milk with simple pretreatment. Conclusion This assay developed can detect all 16 β-1actams demanded by the European Union （EU）. The whole procedure takes only 45 min and can detect 42 samples and the standards with duplicate analysis.

关键词： Penicillin-Binding Protein 2x* β-1actam Multi-residue Milk