Inaugural ASASâCAAV Asia Pacif ic Rim Conference Abstracts

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T215 Induction of systemic immune responses of mice bysubcutaneous route with Lactococcus lactis expressing FaeG. L. Shu-jie,L. Yong-ming, X. Zi-wei*, and W. Yi-cheng, Institution of Husbandry andVeterinary, Zhejiang Academy of Agricultural Science, Hangzhou, ZhejiangProvince, China.Diarrhea of newly weaned piglets caused by enterotoxigenic Escherichia coli(ETEC) is one major cause leading to severe economic losses in the swineindustry. FaeG is the major F4 fimbrial subunit and considered a prospectivevaccine candidate to prevent ETEC infections in piglets. Lactococcus lactis (L.lactis) is a safe antigen delivery vehicle. In this study, we used nisin-controlledgene expression system to produce FaeG and test it antigenicity. The FaeG genewas amplified by PCR from ETEC C83907, inserted into pNZ8112 of L. lactissecretion vector with usp45 signal sequence, and transformed into L. lactisNZ9000. After recombinant stain was induced with 5 ng/mL of nisin for 3 h, thestain pellet and culture supernatant were analyzed by SDS-PAGE. The resultshowed that FaeG protein was detected in the cytoplasm (approximately 27kD) and not detected in the supernatant. The amount of FaeG protein was up to13.56% of the total cellular soluble protein. The Western blot analysis indicatedthat the protein possesses good reactionogenicity. To test recombinant FaeGantigenicity, 30 female BALB/c mice aged 6 wk were divided into 3 groupsfor subcutaneous immunization. One group received 1.5× 10 8 cfu of L. lactisexpressing FaeG, and 2 control groups received PBS or identical quantities ofL. lactis harboring pNZ8112 on d 0, 14, and 35. Blood samples were collectedon d 0 and 49 to evaluate serum IgG response by indirect ELISA with purifiedF4 fimbriae as antigen coated. Mice spleens of each group were removed ond 56 to analyze numbers of F4-specific IgG with purified F4 fimbriae as astimulator by Elispot. The results showed that F4-specific IgG responses ofthe group immunized with recombinant stain was significantly higher thanthe 2 control groups (P < 0.01), and the number of F4-specific IgG cells wasup to 73 per 10 6 MC. The study shows that L. lactis can express FaeG andsubcutaneous immunization with recombinant L. lactis can induce systemicimmune responses in mice.Key Words: FaeG, Lactococcus lactis, subcutaneous inoculationT216 Alleviating effect of Coptis chinensis and berberine onintestinal injury in rats challenged with lipopolysaccharides. Q. Zhang*, T.Lu, D. Wang, and X. Piao, State Key Laboratory of Animal Nutrition, Collegeof Animal Science and Technology, China Agricultural University, Beijing,China.Sepsis is still associated with a high morbidity and mortality rate in recent yearsdespite better supportive therapy and care. In septicemic animals, the barrierfunction of the intestinal mucosa is compromised to aggravate the critical illness.In recent years, Coptis chinensis and its main alkaloid compound, berberine,has been proven to possess anti-inflammatory and antioxidant properties. Thepresent study investigated their effects on endotoxin-induced intestinal mucosaldamage. The yield of C. chinensis aqueous extract (CCAE) was 14% (w/w),and its berberine content was 9.89% determining by high performance liquidchromatography. Forty-eight male Sprague-Dawley rats were divided into fourgroups. Sprague-Dawley rats were orally administrated 30 mg/kg body weightberberine (BBR30), 300 mg/kg body weight aqueous extract of C. chinensis(CCAE300), or the vehicle (saline; 10 ml/kg body weight) every day for2 weeks before being injected with endotoxin (i.p., 20 mg/kg body weight).Control rats were administered and injected with saline. Blood and ileum werecollected 6 h after injection to measure pro-inflammatory cytokine levels byELISA and antioxidant indices by a spectrophotometer. BBR30 and CCAE300treatments lowered tumor necrosis factor-α ± (TNF-α±), interleukin-1α² (IL-1α²) and nitrite oxide (NO) in plasma (P < 0.05), elevated the activities ofsuperoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) (P < 0.05)and decreased the levels of malondialdehyde (MDA) and nitrate/nitrite (P
T219 Stabilization of roxarsone and arsanilic acid infeed storage. J. Wang*, H. Ren, P. Lou, Z. Fu, and J. Wang, College of Bio-Engineering, Henan University of Technology, Zhengzhou, P. R. China.The concentration of inorganic arsenic in feeds is constant, and their enrichmentin animal tissues and organs causes toxic hazards for healthy people. Roxarsoneand arsanilic acid are often supplied in feeds because they have no toxicity in thepermitted dose, but after being stored for a period of time, their speciation maychange. The object of this study was to investigate the stabilization of roxarsoneand arsanilic acid after storage. Complete feeds and concentrate feeds, premixedwith roxarsone or arsanilic acid, were stored at room temperature and at 37°Cwith a moisture content of 10, 12, and 14% for 1, 2, 4, 6, 8, 10, 15, 20, 25, and30 d. The contents of roxarsone and arsanilic acid in feeds were determined byHPLC after storage. Roxarsone levels decreased significantly with an increasein moisture content. At a moisture content of 10, 12, and 14%, roxarsonelevels decreased by 60.5, 75.6, and 80.2%, respectively, at room temperature,and decreased by 70.7, 80.6, and 85.5% at 37°C in the complete feeds. Forconcentrate feeds, roxarsone levels decreased by 80% with different moisturecontents at room temperature and by 80.4, 82.3, and 83.2%, respectively, at37°C. Roxarsone levels decreased by 70.5% at room temperature and by 72.3%at 37°C in the premix. Arsanilic acid levels decreased rapidly within 10 d;nearly 50% of the arsanilic acid in feeds was lost by d 10. The levels decreasedby 55.4% at room temperature and by 52.5% at 37°C in the complete feedsat 30 d, and the moisture in feeds had no effect on arsanilic acid degradation.Arsanilic acid levels were reduced by 88.5% at 37°C and by 59.7% at roomtemperature in concentrate feeds, and the levels decreased by 60.2 and 50.1%in the first 10 d. Arsanilic acid levels in the premix decreased slowly; theywere reduced by 68.0% at 37°C and by 48.5% at room temperature at 1 mo.Roxarsone and arsanilic acid levels in feeds were not stable during storage;arsanilic acid decreased rapidly in 10 d. The arsenic speciation of roxarsone andarsanilic acid degradation may be dangerous for animals.Key Words: roxarsone, arsanilic acid, stabilizationT220 Involvement of ERK1/2 signaling pathway inheat stress-induced damage and expression change of growth factors inrat jejunum and IEC-6 cells. J. Yu* 1,3 , P. Yin 2 , J. Yin 2 , F. Liu 1,3 , X. Zhu 2,3 ,and J. Xu 2,3 , 1 Beijing University of Agriculture, Beijing, P.R. China, 2 ChinaAgricultural University, Beijing, P.R. China, 3 Key Laboratory of Developmentand Evaluation of Chemical and Herbal Drugs for Animal Use, Beijing, P.R.China.Our previous studies showed that small intestine epithelial tissues of animalswere obviously damaged after exposure to severe heat stress and were rapidlyrestored in the following few days. Growth factors, as a critical survival factorto promote endothelial cell proliferation and migration, and the ERK1/2signaling pathway were considered to regulate the growth and adaptation ofendothelial cells to both physiological and pathological stimuli. However, littleinformation is available concerning both the ERK1/2 signaling pathway andgrowth factor expression in heat stress. Thus, we hypothesized that growthfactor messenger RNA (mRNA) expression and the ERK1/2 signaling pathwaywere involved in the heat stress-induced damage and regeneration process. Totest this hypothesis, 48 male Sprague-Dawley rats were randomly assigned toeither a control group or a heat-stressed group. Rats in the control group weremaintained at 25°C, 60% relative humidity, whereas rats in the heat-stressedgroup were exposed to 40°C for 2 h, once a day, for 10 consecutive days. On d1, 3, 6, and 10, six rats from each group were killed and sampled immediatelyat the end of heat stress. Rat IEC-6 cells were assigned to a control group and aheat-stressed group. The control group cells were maintained at 37°C, 5% CO 2,whereas cells from the heat-stress group were exposed to 42°C for 3 h. Heatstress caused morphology damage to the rat jejunum and IEC-6 cells in thehistological assay, reduced cell proliferation viability in the MTT assay, inducedcell apoptosis in the FACS assay, changed growth factor mRNA expression inboth the microarray and real-time PCR, and induced activation of the ERK1/2signaling pathway in the Western blot assay. Heat stress caused more severemorphology damage and cell apoptosis, and affected growth factor mRNAexpression when the ERK1/2 signaling pathway was inhibited by U0126. Inconclusion, both the ERK1/2 signaling pathway and growth factor mRNAexpression were involved in heat stress. The activation of the ERK1/2 signalingpathway acted as a survival mechanism via regulated growth factor expressionin heat-stressed IEC-6 cells.T221 The study of volatile oil of garlic on the effect oftumor necrosis factor-α and immunoglobulin A changes in mice infected byEscherichia coli. G. Cheng* 1,3 , F. Liu 1,3 , and J. Xiu 2 , 1 Beijing Key Laboratoryof TCVM, Beijing, China, 2 Beijing University of Agriculture, Beijing, China,3China Agricultural University, Beijing, China.The study initially discussed a sterilization mechanism through an in vitroantibacterial experiment with the traditional Chinese medicine volatile oil ofgarlic, then duplicated the mouse colibacillosis model successfully, and furtherdiscussed the influence of volatile oil of garlic on serum IgA and TNF-αon the basis of the model. From the duplication level, we studied the RNAexpression of IgA and TNF-α in the ilea of a colibacillosis-infected mouse fedvolatile oil of garlic. The results are as follows. 1) In an in vitro antibacterialexperiment, the effect of GO-II primarily by DADS was obviously better thanthe effect of GO-I primarily by DATS, and the effect on piglet colibacteriawas on the order DADS > GO-II > GO-I = DATS. We discovered, througha time passage sterilization curve, that the volatile oil of garlic is one kind ofdensity-dependent medicine. 2) The colibacillosis-infected mouse model wassuccessfully duplicated by a standard strain of piglet colibacteria (CVCC210),and the clinical symptoms and change in histology of the mouse model talliedwith the pig colibacillosis. 3) The volatile oil of garlic effectively suppressedNOS activity and NO production in the ileal mucous membrane organizationinduced by LPS, which is produced by colibacteria. 4) The volatile oil of garliceffectively suppressed the TNF-α content in blood serum and enhanced thecontent of IgA (P < 0.05) in the model group. 5) The quantity of TNF-α and IgAmessenger RNA expression in the ileal mucous membrane was consistent withthe quantity expressed in blood serum in the volatile oil of garlic group. Thesecretion level in mouse blood serum and the TNF-α IgA in the small intestinalmucous membrane could be adjusted and restored to the normal levels in thevolatile oil of garlic group.Key Words: Escherichia coli, garlic volatile oil, tumor necrosis factor-αT222 Use of Caenorhabditis elegans as an animal model toevaluate Lactobacillus isolates for use as probiotics to control SalmonellaTyphimurium. W. Chunyang 1,2 , G. Joshua* 2 , N. Zhongxiang 1 , Y. Hai 2 , and A.Hawke 2 , 1 Shandong Agriculture University, Tan’an, Shandong,China, 2 GuelphFood Research Centre, Agriculture and Agri-Food Canada, Guelph, Ontario,Canada.Salmonella Typhimurium is a virulent pathogen for humans and animals. Thisstudy was undertaken to test Caenorhabditis elegans for its suitability as an invivo model for screening useful probiotic bacteria and to select isolates that canbe further evaluated by trials with food animals for the development of effectiveprobiotics for Salmonella control. Caenorhabditis elegans strain SS104 wasused. Antimicrobial activity was estimated through the “spot-on-the-lawn”assay. Antimicrobial activity of the extracellular culture fluid was tested by acoculture experiment. Assay for the life span of C. elegans was conducted anda large adjustment was done. After 3 washes in S medium by centrifugationto pellet and resuspend worms, the nematode was kept in 24-well titer plateswith each well containing 2 mL of S medium and 10 to 15 worms, and wasincubated at 25°C during the assay. Different treatments were designed. Weevaluated 17 Lactobacillus isolates for their ability to protect the nematodefrom Salmonella-induced death. These isolates were inhibitive to SalmonellaTyphimurium but showed no significant difference in our microbiologicalassays for inhibition towards the pathogen. When tested on C. elegans infectedwith Salmonella Typhimurium, however, the isolates were differentiatedin their protection. Although 4 of the isolates showed no or little protection,protection of the remaining isolates varied in degree, with 5 of them providingfull or nearly full protection to the nematode. Our observations suggest that C.elegans can be used as a prescreening animal model to study the efficacy ofprobiotics to inhibit intestinal Salmonella infection. The selected Lactobacillusisolates warrant further study with Salmonella-challenged chickens or pigs forthe development of effective probiotic agents.Key Words: Lactobacillus, Caenorhabditis elegans, Salmonella TyphimuriumKey Words: heat stress, ERK1/2, growth factor63
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Inaugural ASAS-CAAVAsia Pacif ic Ri
Page 3 and 4:
Scientific ProgramTable of Contents
Page 5 and 6:
1 Advanced needle-free injection te
Page 7 and 8:
9 Pig personality, meat quality, an
Page 9 and 10:
17 The contamination and distributi
Page 11 and 12: 25 Genetic evaluations for measures
Page 13 and 14: of control and the lowest of SDAP g
Page 15 and 16: 39 Effects of bacterial protein and
Page 17 and 18: Advances in Digestive Physiology Me
Page 19 and 20: L-arginine increased (P < 0.05) the
Page 21 and 22: average final weight (AFW) and aver
Page 23 and 24: 71 Building a foundation: Cells, st
Page 25 and 26: 78 Effect of the level of vitamin A
Page 27 and 28: 86 Evaluation of phosphorus excreti
Page 29 and 30: 94 Responses of dairy cows to suppl
Page 31 and 32: 102 Construction and analysis of a
Page 33 and 34: M132 Study on the effects of pectin
Page 35 and 36: M140 Effect of Mintrex Zn on perfor
Page 37 and 38: M148 Effect of the hydrolyzed wheat
Page 39 and 40: treatment 1 was significantly lower
Page 41 and 42: M163 The main fatty acid contents i
Page 43 and 44: M170 Zinc requirements of yellow br
Page 45 and 46: M178 Influences of dietary riboflav
Page 47 and 48: M185 Application of an advanced syn
Page 49 and 50: M193 Studies on the effects of oreg
Page 51 and 52: M202 Plasma leucine turnover rate,
Page 53 and 54: 103 Use of natural antimicrobials t
Page 55 and 56: 111 The somatotropic axis in growth
Page 57 and 58: Environmental Impacts of Cattle, Sw
Page 59 and 60: 128 Opportunities for international
Page 61: Animal Health PostersT211 Locoweed
Page 65 and 66: Beef Species PostersUrinary purine
Page 67 and 68: T233 The effects of sire and breed
Page 69 and 70: T242 Ultrastructure of oocyte and e
Page 71 and 72: T249 Effect of different combinatio
Page 73 and 74: Forages and Pastures PostersIn vitr
Page 75 and 76: T263 Effects of leaf meal of Brouss
Page 77 and 78: T271 The effects of feeding expandi
Page 79 and 80: Lactation Biology PostersT278 Effec
Page 81 and 82: Physiology and Endocrinology Poster
Page 83 and 84: T288 Effect of Aspergillus meal pre
Page 85 and 86: Poultry Physiology, Endocrinology,
Page 87 and 88: T301 Observation of the feeding man
Page 89 and 90: T307 Effect of levels of Yucca schi
Page 91: T313 Study of lysine requirement of
Page 94 and 95: energy, 5, 26energy and nutrient di
Page 96 and 97: protein digestive enzyme, 44protein
Page 98 and 99: HHai, Y., T222, T248Hai-Ying, Z., T
Page 100 and 101: Song, X., T223Song, Z. G, M144, T20
Page 102: 102NOTES
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