作者机构：
[Li, Chuanfeng; Fu, Yuzhi; Chen, Zongyan; Liu, Guangqing] Chinese Acad Agr Sci, Shanghai Vet Res Inst, Shanghai 200241, Peoples R China.;[Fu, Yuzhi] Hunan Agr Univ, Anim Med Coll, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[Liu, Guangqing] C;Chinese Acad Agr Sci, Shanghai Vet Res Inst, Shanghai 200241, Peoples R China.
关键词：
Suicidal DNA vaccine;Duck hepatitis virus;VP1 gene;Ducklings
摘要：
A suicidal DNA vaccine based on a Semliki Forest virus (SFV) replicon was evaluated for the development of a vaccine against duck hepatitis virus type 1 (DHV-1). The VP1 gene of DHV-1 was cloned and inserted into pSCA1, an SFV DNA-based replicon vector. The resultant plasmid, pSCA/VP1, was transfected into BHK-21 cells and the antigenicity of the expressed protein was confirmed using an indirect immunofluorescence and western blot assay. Immunogenicity was studied in ducklings. Ducklings were injected intramuscularly two times with pSCA/VP1 at 14 days intervals. Anti-DHV-1 antibodies were detected by ELISA, the lymphocyte proliferation response was also tested by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide method and neutralizing antibodies were measured by microneutralization tests. Our results showed that DHV-1-specific antibodies, neutralizing antibodies and lymphocyte proliferation were well induced in ducklings. Furthermore, all the ducklings were protected against challenge with wild DHV-1. In conclusion, we demonstrate that the suicidal DNA vaccine is a promising vaccine candidate facilitating the prevention of duck hepatitis caused by DHV-1.
作者机构：
[Wang, D.] Fudan Univ, Shanghai Med Coll, Key Lab Med Mol Virol, Shanghai 200032, Peoples R China.;[She, R.; Liu, Y.] China Agr Univ, Coll Vet Med, Beijing 100193, Peoples R China.;[Jia, X.] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[She, R.] C;China Agr Univ, Coll Vet Med, Beijing 100193, Peoples R China.
关键词：
eosinophilic major basic protein;mast cell;very virulent infectious bursal virus;eotaxin
摘要：
Very virulent infectious bursal disease virus (vvIBDV) can cause systemic inflammatory syndromes and acute death in specific-pathogen-free chickens within 72 h. However, the subtle mechanism of these severe inflammatory responses has been unsatisfactorily resolved until now. This study determined the kinetics of mast cells, tryptase, eosinophilic major basic protein, and eotaxin expression in specific-pathogen-free chickens after vvIBDV infection. Results showed that mast cell population, tryptase activity, major basic protein, and eotaxin expression were increased markedly in the vvIBDV-infected animals compared with the controls, with a significant difference in the bursa. Acute inflammatory lesions and high mortality were observed in vvIBDV-infected chickens. These observations implicate activated mast cells and eosinophils as important participants in vvIBDV-induced acute inflammatory lesions through mediators released in a short timeframe.
摘要：
The prevalence of porcine sapovirus infection in weanling pigs was investigated in Hunan Province, China, between August 2006 and October 2007. A total of 153 diarrheic fecal samples from ten intensive pig farms from ten representative administrative regions in Hunan province were examined for porcine sapoviruses using RT-PCR. Twenty-two of 153 (14.37 %) samples were found to contain porcine sapoviruses. Phylogenetic analysis showed that all the porcine sapovirus isolates in Hunan Province belonged to the porcine sapovirus genogroup III. The results of the present investigation have implications for the control of porcine sapovirus infection in pigs in Hunan Province, China.
作者机构：
[Liu, G. H.; Cai, X. Q.; Song, H. Q.; Wu, C. Y.; Lin, R. Q.; Zhu, X. Q.] CAAS, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou 730046, Gansu, Peoples R China.;[Cai, X. Q.] Zhongshan Entry Exit Inspect & Quarantine Bur, Zhongshan 528403, Guangdong, Peoples R China.;[Liu, G. H.; Zhu, X. Q.] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Zou, F. C.; Zhu, X. Q.] Yunnan Agr Univ, Coll Anim Sci & Technol, Kunming 650201, Yunnan Province, Peoples R China.;[Yan, H. K.; Lin, R. Q.; Yuan, Z. G.] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China.
通讯机构：
[Lin, R. Q.] C;CAAS, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou 730046, Gansu, Peoples R China.
关键词：
Schistosoma;Liver Fluke;Schistosoma Japonicum;Clonorchiasis;High Statistical Support
作者机构：
[Wu, Yong; Sun, Zhi-Liang; Liu, Zhao-Ying] Hunan Agr Univ, Coll Vet Med, Hunan Engn Res Ctr Vet Drug, Changsha 410128, Hunan, Peoples R China.;[Wan, Leren] Shimadzu Int Trading Shanghai Co Ltd, Shanghai 200052, Peoples R China.
通讯机构：
[Liu, Zhao-Ying] H;Hunan Agr Univ, Coll Vet Med, Hunan Engn Res Ctr Vet Drug, Changsha 410128, Hunan, Peoples R China.
关键词：
Diaveridine;Hybrid ion trap/time-of-flight mass spectrometry;Metabolism;Pig liver microsomes;Trimethoprim
摘要：
Trimethoprim (TMP) and diaveridine (DVD) are used in combination with sulfonamides and sulfaquinoxlaine as an effective antibacterial agent and antiprotozoal agent, respectively, in humans and animals. To gain a better understanding of the metabolism of TMP and DVD in the food-producing animals, the metabolites incubated with liver microsomes of pigs were analyzed for the first time with high-performance liquid chromatography combined with hybrid ion trap/time-of-flight mass spectrometry. Seven TMP-related and six DVD-related metabolites were characterized based on the accurate MS2 spectra and known structure of the parent drug, respectively. The metabolites of TMP were identified as two O-demethylation metabolites, a di-O-demethylation metabolite, two N-oxides metabolites, a hydroxylated metabolite on the methylene carbon and a hydroxylated metabolite on the methyl group. DVD was also biotransformed to two O-demethylation metabolites, a di-O-demethylation metabolite, an N-oxide metabolite, a hydroxylation metabolite on the methylene carbon and a hydroxylation metabolite followed by O-demethylation. The results indicate that the two compounds have similar biotransformation pathways in pigs. O-Demethylation was the major metabolic route of TMP and DVD in the pig liver microsomes. The proposed metabolic pathways of TMP and DVD in liver microsomes will provide a basis for further studies of the in vivo metabolism of the two drugs in food-producing animals. Copyright (c) 2011 John Wiley & Sons, Ltd.
作者机构：
[Song, Hui-Qun; Liu, Guo-Hua; Zhu, Xing-Quan; Hou, Jie] Chinese Acad Agr Sci, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou 730046, Gansu, Peoples R China.;[Liu, Guo-Hua; Zhu, Xing-Quan] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Yuan, Zi-Guo; Lin, Rui-Qing; Li, Shu; Weng, Ya-Biao; Hou, Jie] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China.;[Zhu, Xing-Quan] Yunnan Agr Univ, Coll Anim Sci & Technol, Kunming 650201, Yunnan Province, Peoples R China.
通讯机构：
[Zhu, Xing-Quan] C;Chinese Acad Agr Sci, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou 730046, Gansu, Peoples R China.
作者机构：
[Song, Hui-Qun; Xu, Min-Jun; Huang, Si-Yang; Wu, Chang-Yi; Liu, Guo-Hua; Zhu, Xing-Quan] CAAS, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou 730046, Gansu, Peoples R China.;[Liu, Guo-Hua; Zhu, Xing-Quan] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Song, Hui-Qun; Wu, Chang-Yi; Lin, Rui-Qing] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China.;[Wei, Shu-Jun] Beijing Acad Agr & Forestry Sci, Inst Plant & Environm Protect, Beijing 100097, Peoples R China.;[Zhao, Guang-Hui] NW A&F Univ, Coll Vet Med, Yangling 712100, Shaanxi Provinc, Peoples R China.
通讯机构：
[Zhu, Xing-Quan] C;CAAS, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou 730046, Gansu, Peoples R China.
作者：
Liu, G. H.*;Li, B.;Li, J. Y.;Song, H. Q.;Lin, R. Q.;...
期刊：
JOURNAL OF HELMINTHOLOGY,2012年86(4):479-484 ISSN：0022-149X
通讯作者：
Liu, G. H.
作者机构：
[Liu, G. H.; Li, B.; Li, J. Y.; Song, H. Q.; Zhou, D. H.; Zhu, X. Q.] Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol, Lanzhou 730046, Gansu, Peoples R China.;[Liu, G. H.; Zhu, X. Q.] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Li, B.] Guangdong HuanongWens Anim Husb Co Ltd, Yunfu 527400, Guangdong, Peoples R China.;[Li, B.; Li, J. Y.; Yan, H. K.; Lin, R. Q.; Yuan, Z. G.] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China.;[Cai, X. Q.] Zhongshan Entry Exit Inspect & Quarantine Bur, Zhongshan 528403, Guangdong, Peoples R China.
通讯机构：
[Liu, G. H.] C;Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol, Lanzhou 730046, Gansu, Peoples R China.
摘要：
The present study examined sequence variation in four mitochondrial (mt) genes, namely cytochrome c oxidase subunits 1 (cox1) and 2 (cox2), and NADH dehydrogenase subunits 1 and 2 (nad1 and nad2) among Clonorchis sinensis isolates from different endemic regions in China, and their phylogenetic relationships with other zoonotic trematodes were reconstructed. A portion of the cox1 and cox2 genes (pcox1 and pcox2), and nad1 and nad2 genes (pnad1 and pnad2) were amplified separately from individual liver flukes by polymerase chain reaction (PCR) and the amplicons were subjected to sequencing from both directions. The intra-specific sequence variations within C. sinensis were 0-1.6% for pcox1, 0-1.4% for pcox2, 0-0.9% for pnad1 and 0-1.0% for pnad2. Phylogenetic analyses based on the combined sequences of pcox1, pcox2, pnad1 and pnad2 revealed that all the C. sinensis isolates grouped together and were closely related to Opisthorchis felineus. These findings revealed the existence of intra-specific variation in mitochondrial DNA (mtDNA) sequences among C. sinensis isolates from different geographic regions, and demonstrated that mtDNA sequences provide reliable genetic markers for phylogenetic studies of zoonotic trematodes.
摘要：
The seroprevalence of porcine cytomegalovirus (PCMV) and sapovirus (SaV) infections in pigs was investigated in Hunan province, China, between May 2005 and October 2010. A total of 500 pig serum samples collected from 10 representative administrative regions in Hunan province were evaluated for antibodies against PCMV and SaV using enzyme-linked immunosorbent assay (ELISA). The overall seroprevalence of porcine cytomegalovirus and sapovirus in pigs was 96.40% (482/500) and 63.40% (317/500), and the seropositivity of 10 herds we surveyed varied, ranging from 94.74% to 98.48% and 56.36% to 72.50%, respectively. The highest prevalence was found in breeding sows (96.67% for PCMV and 83.33% for SaVs). The results of the present survey indicated that infections with porcine cytomegalovirus and sapovirus are highly prevalent in pigs in Hunan province, China.
期刊：
POLISH JOURNAL OF VETERINARY SCIENCES,2012年15(2):305-313 ISSN：1505-1773
通讯作者：
Obminska-Mrukowicz, B.
作者机构：
[Jine, Y.] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Szczypka, M.; Lis, M.; Obminska-Mrukowicz, B.] Wroclaw Univ Environm & Life Sci, Dept Biochem Pharmacol & Toxicol, Fac Vet Med, PL-50375 Wroclaw, Poland.;[Obminska-Mrukowicz, B.] Wroclaw Univ Environm & Life Sci, Dept Biochem Pharmacol & Toxicol, Fac Vet Med, Norwida 31, PL-50375 Wroclaw, Poland.
通讯机构：
[Obminska-Mrukowicz, B.] W;Wroclaw Univ Environm & Life Sci, Dept Biochem Pharmacol & Toxicol, Fac Vet Med, Norwida 31, PL-50375 Wroclaw, Poland.
关键词：
Life Sciences;Zoology;Life Sciences, other;Medicine;Veterinary Medicine
摘要：
Betulinic acid is a pentacyclic triterpene found in many plant species, among others, in the bark of white birch Betula alba. Betulinic acid was reported to display a wide range of biological effects, including antiviral, antiparasitic, antibacterial, anticancer and anti-inflammatory activities. The effects of betulinic acid (50, 5, 0.5 mg/kg) administered orally five times at 24 hours intervals to non-immunized and red blood cells (SRBC)-immunized mice were determined. The present study examined the total number of lymphocytes in the thymus, spleen and mesenteric lymph nodes, and the percentage of subsets of T cells (CD4(+)CD8(+), CD4(-)CD8(-), CD4(+), CD8(+)) in thymus,T (CD3(+), CD4(+), CD8(+)) and B (CD19(+)) lymphocytes in the spleen and mesenteric lymph nodes, as well as white blood cell (WBC) and differential leukocyte counts in non-immunized mice, and humoral immune response in SRBC-immunized mice. SRBC was injected 24 hours after administration of the last dose of betulinic acid. It was found that betulinic acid administered orally five times at the dose of 0.5 mg/kg increased the total number of thymocytes, splenocytes, lymphocytes of mesenteric lymph node cells, and the weight ratio of the spleen and mesenteric lymph nodes in non-immunized mice. Betulinic acid also changed the percentage of T cell subsets in the thymus and T and B lymphocytes in peripheral lymphatic organs. The effects of betulinic acid on T and B cell subpopulations depended on the dose applied. The strongest stimulating effect of betulinic acid was observed when the drug was administered at the dose of 0.5 mg/kg. Five exposures to betulinic acid (0.5 mg/kg) decreased the percentage of immature CD4(+)CD8(+) thymic cells with corresponding increases in the percentage and absolute count of mature, single-positive CD4(+) thymocytes and decreased the percentage and total count of CD3(+) splenocytes and mesenteric lymph node cells with corresponding decreases in the percentage and absolute count of CD4(+) and CD8(+) cells. Multiple administration of betulinic acid at the investigated doses augmented the percentage and absolute count of CD19(+) cells in the peripheral lymphatic organs. Moreover, betulinic acid at the dose of 5 mg/kg administered prior to SRBC immunization increased the number of plaque forming cells (PFC) but decreased the production of anti-SRBC antibodies on day 4 after priming. Thus, betulinic acid is a potential biological response modifier and may strengthen the immune response of its host.
期刊：
Research in Veterinary Science,2012年93(2):710-715 ISSN：0034-5288
通讯作者：
Hu, S.
作者机构：
[Yang, X.; Ding, J.; Wang, G.; Hu, S.] Hunan Agr Univ, Fac Vet, Changsha, Hunan, Peoples R China.
通讯机构：
[Hu, S.] H;Hunan Agr Univ, Fac Vet, Changsha, Hunan, Peoples R China.
关键词：
Avian influenza virus;Enzyme-linked immunosorbent assay;Matrix 1 protein
摘要：
Fragment of 759 bp DNA spanning the Matrix 1 (M1) gene of Avian Influenza Virus (AIV) was inserted into an expression vector pET28c to construct a recombinant plasmid pET28c-M1. The pET28c-M1 plasmid was transformed into the Escherichia coli BL21 (DE3) competent cell to produce a recombinant strain E. coli 21 (DE3). After being induced by Isopropyl-b-D-galactopyranoside (IPTG), E. coli 21 (DE3) expressed a 28-kDa fusion protein at a high level. This protein can bind anti-AIV (H5N1) positive serum by Western-blot analysis. After being denatured, renatured, and purified by Ni2+-column, the fusion protein was used as an antigen to develop Matrix 1 Enzyme-Linked lmmunosorbent Assay (M1-ELISA) for detecting antibodies against AIV from chicken serum. We found that this indirect M1-ELISA was sensitive for differentiating antisera against AIV and antisera against other six kinds of avian viruses apart from AIV and this method is more sensitive than Hemagglutination Inhibition (HI) test. When compared with HI test and ELISA (IDEXX) in evaluating 581 serum samples from field vaccinated chickens, this assay showed 93.3% agreement ratio with the HI test, as well as 96.0% agreement ratio with ELISA (IDEXX). In a preliminary application, the assay successfully detected 19 AIVs from 51 nonvaccinated chicken lungs. It concludes that an indirect ELISA was successfully developed for detecting AIV. The assay is specific and sensitive. The application will greatly contribute to the long-term prevention and control of avian influenza in China. (c) 2011 Elsevier Ltd. All rights reserved.
作者机构：
[Song, Hui-Qun; Lin, Rui-Qing; Zhang, Yuan; Liu, Guo-Hua; Zhu, Xing-Quan; Zhou, Dong-Hui] Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol, Lanzhou 730046, Gansu, Peoples R China.;[Song, Hui-Qun; Yuan, Zi-Guo; Lin, Rui-Qing; Zhang, Yuan] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China.;[Liu, Guo-Hua; Zhu, Xing-Quan] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[D'Amelio, Stefano] Sapienza Univ Rome, Parasitol Sect, Dept Publ Hlth & Infect Dis, I-00185 Rome, Italy.;[Zhu, Xing-Quan; Zou, Feng-Cai] Yunnan Agr Univ, Coll Anim Sci & Technol, Kunming 650201, Yunnan Province, Peoples R China.
通讯机构：
[Zhu, Xing-Quan] C;Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol, Lanzhou 730046, Gansu, Peoples R China.
作者机构：
[Song, Hui-Qun; Xu, Min-Jun; Wang, Shu-Yan; Zhao, Guang-Hui; Liu, Guo-Hua; Zhu, Xing-Quan; Zhou, Dong-Hui] Chinese Acad Agr Sci, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou, Gansu, Peoples R China.;[Liu, Guo-Hua; Zhu, Xing-Quan] Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.;[Wang, Shu-Yan; Huang, Wei-Yi] Guangxi Univ, Coll Anim Sci & Technol, Nanning 530004, Guangxi Zhuang, Peoples R China.;[Zhao, Guang-Hui] NW A&F Univ, Coll Vet Med, Yangling, Shaanxi Provinc, Peoples R China.;[Wei, Shu-Jun] Beijing Acad Agr & Forestry Sci, Inst Plant & Environm Protect, Beijing, Peoples R China.
通讯机构：
[Zhu, Xing-Quan] C;Chinese Acad Agr Sci, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou, Gansu, Peoples R China.
关键词：
Transfer RNA;Invertebrate genomics;Genomics;Phylogenetic analysis;Gastropods;Mammalian genomics;Mitochondrial DNA;Mitochondria
摘要：
Complete mitochondrial (mt) genomes and the gene rearrangements are increasingly used as molecular markers for investigating phylogenetic relationships. Contributing to the complete mt genomes of Gastropoda, especially Pulmonata, we determined the mt genome of the freshwater snail Galba pervia, which is an important intermediate host for Fasciola spp. in China. The complete mt genome of G. pervia is 13,768 bp in length. Its genome is circular, and consists of 37 genes, including 13 genes for proteins, 2 genes for rRNA, 22 genes for tRNA. The mt gene order of G. pervia showed novel arrangement (tRNA-His, tRNA-Gly and tRNA-Tyr change positions and directions) when compared with mt genomes of Pulmonata species sequenced to date, indicating divergence among different species within the Pulmonata. A total of 3655 amino acids were deduced to encode 13 protein genes. The most frequently used amino acid is Leu (15.05%), followed by Phe (11.24%), Ser (10.76%) and IIe (8.346%). Phylogenetic analyses using the concatenated amino acid sequences of the 13 protein-coding genes, with three different computational algorithms (maximum parsimony, maximum likelihood and Bayesian analysis), all revealed that the families Lymnaeidae and Planorbidae are closely related two snail families, consistent with previous classifications based on morphological and molecular studies. The complete mt genome sequence of G. pervia showed a novel gene arrangement and it represents the first sequenced high quality mt genome of the family Lymnaeidae. These novel mtDNA data provide additional genetic markers for studying the epidemiology, population genetics and phylogeographics of freshwater snails, as well as for understanding interplay between the intermediate snail hosts and the intra-mollusca stages of Fasciola spp..
通讯机构：
[Yu, Xinglong] H;Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.
摘要：
A double-antigen sandwich enzyme-linked immunosorbent assay (ELISA) is described for detection of porcine circovirus 2 (PCV2) antibodies using the well-characterized recombinant PCV2 capsid protein. In a comparative test of 394 pig sera against an indirect immunofluorescence (IIF) test and a commercial ELISA kit (also based on the recombinant PCV2 capsid protein), the results showed that the diagnostic sensitivity, specificity, and accuracy of the assay were, respectively, 90.61, 94.02, and 91.62% compared with IIF and 94.38, 95.28, and 94.67% compared with the commercial ELISA kit. Assay of 12 PCV-free pigs over a 5-week period produced only PCV2-negative titers by all 3 methods. These results and the seroprofiles of 4 pig farms obtained by both the commercial ELISA kit and the double-antigen sandwich ELISA indicate that the sandwich ELISA is a reliable method for detection of antibodies to PCV2. Additionally, the method described here permits the use of undiluted test serum samples simultaneously loaded with horseradish peroxidase (HRP)-conjugated antigen into the test well, and the complete test procedure can be performed in less than 90 min. This double-antigen sandwich ELISA should be a useful tool to aid swine industry professionals in deciding the intervention strategies for the control of PCV2-associated diseases.
作者：
Dai, R. S.*;Liu, G. H.;Song, H. Q.;Lin, R. Q.;Yuan, Z. G.;...
期刊：
JOURNAL OF HELMINTHOLOGY,2012年86(2):245-251 ISSN：0022-149X
通讯作者：
Dai, R. S.
作者机构：
[Liu, G. H.; Liu, W.; Dai, R. S.] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Liu, G. H.; Song, H. Q.; Liu, W.; Lin, R. Q.; Huang, S. Y.; Dai, R. S.; Zhu, X. Q.] CAAS, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou 730046, Gansu, Peoples R China.;[Song, H. Q.; Lin, R. Q.; Yuan, Z. G.] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China.;[Li, M. W.] Guangdong Ocean Univ, Dept Vet Med, Zhanjiang 524088, Guangdong, Peoples R China.;[Zhu, X. Q.] Yunnan Agr Univ, Coll Anim Sci & Technol, Kunming 650201, Yunnan, Peoples R China.
通讯机构：
[Dai, R. S.] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
作者机构：
[Li, Zhong-Yuan; Chen, Jia; Liu, Guo-Hua; Zhu, Xing-Quan; Zhou, Dong-Hui] Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol, Lanzhou 730046, Gansu, Peoples R China.;[Li, Zhong-Yuan; Chen, Jia] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China.;[Liu, Guo-Hua; Zhu, Xing-Quan] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[Zhu, Xing-Quan] C;Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol, Lanzhou 730046, Gansu, Peoples R China.
摘要：
Toxoplasma gondii is a highly prevalent protozoan parasite infecting a wide range of animals and humans. The epidemiological and biological diversity of T. gondii has resulted in a high genetic variation and unusual population structure in this parasite. This study examined sequence diversity in dense granule 5 (GRA5) gene among T. gondii isolates from different hosts and geographical regions. The entire genome region of the GRA5 gene was amplified and sequenced from 14 T. gondii isolates, and phylogenetic relationship among these T. gondii isolates was reconstructed using Bayesian inference (BI) and maximum parsimony (MP) based on the GRA5 sequences. The complete sequence of the GRA5 gene was 1614 bp in length for strains TgCatBr5 and MAS, but 1617 bp for the other 12 strains. Sequence analysis identified 41 (0–1.7%) variable nucleotide positions among all isolates, with 18 variations of these being in the coding region. Variable positions in the coding region resulted in 11 amino acid substitutions, and a deletion of 3 bp in the strains TgCatBr5 and MAS leading to the deletion of one amino acid. Sequence variations resulted in the existence of polymorphic restriction sites for endonucleases Aat II and Mlu I, allowing the differentiation of the three major clonal lineage types I, II and III by PCR-RFLP. Phylogenetic analyses using BI and MP supported the clear differentiation of the examined T. gondii strains into their respective genotypes. This study demonstrated the existence of sequence variability in the GRA5 gene sequence among T. gondii isolates from different hosts and geographical regions, which allowed the differentiation of the examined T. gondii strains into their respective genotypes, suggesting that this highly polymorphic GRA5 locus may provide a new genetic marker for population genetic studies of T. gondii isolates.
摘要：
Zearalenone is a mycotoxin produced mainly by Fusarium. There are numerous incidences of mycotoxicosis in laboratory and domestic animals, especially in pigs. However, little is known about molecular mechanisms of zearalenone toxicity. Granulosa cells are the maximal cell population in follicles, and they play an essential role in the development and maturation of follicles. The objective of this study was to explore the effect of zearalenone at high concentrations on proliferation and apoptosis of porcine granulosa cells and uncover signaling pathway underlying the cytotoxicity of zearalenone. We found that zearalenone reduced the proliferation of porcine granulosa cells in a dose-dependent manner as shown by the MTT assay and zearalenone resulted in an obvious apoptosis and necrosis in porcine granulosa cells as determined by the TUNEL analysis and flow cytometry. In addition, TUNEL assay with caspase inhibitors showed that zearalenone triggered a caspase-3- and caspase-9-dependent apoptotic process in porcine granulosa cells. Fluorescence spectrophotometer displayed that zearalenone led to a loss of mitochondrial transmembrane potential of porcine granulosa cells but enhanced reactive oxygen species (ROS) levels of the cells. Notably, Western blots revealed that caspase-3 and caspase-9 were activated by zearalenone in porcine granulosa cells. Collectively, our results suggest that zearalenone induces apoptosis and necrosis of porcine granulosa cells in a dose-dependent manner via a caspase-3- and caspase-9-dependent mitochondrial pathway. This study thus offers a novel insight into molecular mechanisms by which zearalenone has adverse cytotoxicity on reproduction. J. Cell. Physiol. 227: 18141820, 2012. (C) 2011 Wiley Periodicals, Inc.
