Acta Veterinaria et Zootechnica Sinica

Mapping of Porcine DIO3 Gene and Analysis of Its Potential Influence on Production Traits

JIANG Xiaoling;ZHAO Xiaofeng;GUO Xiaoling;XU Ningying

2010, 41(4): 383-386. doi:

Abstract ( 636 )

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To investigate if porcine DIO3 gene could be used as a candidate gene influencing some production traits of porcine, porcine DIO3 gene specific primers were designed and used in the radiation hybrid mapping. The RH mapping result showed that this gene was near to the microsatellite SW764 on chromosome 7. Seven QTLs for muscular fiber diameter, internal fat percentage, carcass length, carcass weight, and cortisol level, respectively, were found at this locus by searching pig QTL database. The results suggest that DIO3 could be a candidate gene for porcine meat quality traits, carcass traits and stress response.

Identifying ESTs Differential Expressed in Liver Organ of BlackBone Silky Fowl by Using Differential Displayed PCR

LI Liang;ZHAO Xiaoling;LI Qin;ZHU Qing

2010, 41(4): 387-391. doi:

Abstract ( 1020 )

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In order to find out genes related to special characters of the BlackBone Silky Fowl, differential display PCR was used for screening differential expressed genes between liver organs of the Silkies and the AA broilers, and bioinformatics analysis were carried out to identify ESTs differential expressed. There were totally 10 ESTs (Acc_N: CN606328CN606329, CN606331CN606338) screened from the Silkies′and the AA broilers′liver organs. Results of bioinformatics analysis showed, among 10 ESTs, CN606332 and CN606336 were part of the chicken 18S rRNA gene; CN606333 was homologous with the chicken heat shock 70 ku protein 5 (HSPA5) gene; CN606328 and CN606331 were significant similar with some of chicken cDNA clones or ESTs but had unknown function; the others had no significant similar with any chicken nucleotide sequences but had partly similarity to other species′genes, including genes of membrane protein, gammaglutamyl transpeptidase, ATPbinding cassette transporter, putative protein MGC27019, and abnormal spindle microcephaly protein (ASPM). It was suggested in this study that many genes on varieties of metabolism pathways were differentially expressed between the Silkies and broilers. To study these genes more deeply could provide important information for studying the formation of special characters of the Silkies.

Cloning and Sequence Analysis of the Bovine GDF5 Gene Promoter

LIU Yong-feng;ZAN Lin-sen;;ZHAO Shuan-ping;;QU Kai-xing;LI Lin-qiang;ZHANG Ying-ying;TANG Zhong-lin;YANG Shu-lin;MU Yu-lian;CUI Wen-tao

2010, 41(4): 392-397. doi:

Abstract ( 763 )

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This experiment was conducted to study the potential regulatory sequences of growth differentiate factor 5（GDF5）. The 5′ flanking region of bovine GDF5 gene was amplified through comparing human GDF5 gene, and then a 2 043 bp promoter sequence was confirmed after product purification, ligation, transformation and sequence. In addition, considering the confirmed human GDF5 promoter structure and the analytic result of promoter online software, the promoter sequence was analyzed. It was found that there are no CpG island existing in 5′ flanking region of bovine GDF5 gene using CpG Island Searcher program, and comparing with the human GDF5 5′ flanking sequence, about 78% nucleotide identity was showed. Meanwhile, the result showed that no typical TATA or CAAT boxes were found in bovine GDF5 promoter, and the transcription start site was mapped to -359 bp of translation start site, and the potential transcription factor binding motifs were predicted, including AMLla, Ap1, AmLla, CdxA, CdxA, TATA, AmLla, GTATA1, MZF1, CdxA, Nkx2, CdxA, S8 and SRY, in which only Ap1, AmLla, Nkx2, S8 and SRY were located at the identical positions in human and bovine, both of which were conserved. Moreover, it also showed that around the promoter region from －457 to －423 bp the repeat of GT could increase the promoter activity and a minimal enhancer element reside from －458 to －377 bp in bovine GDF5 promoter. In this experiment, we inferred the transcription start site, transcription factor binding motifs, activity sequence and minimal enhancer element in bovine GDF5 promoter, which may be helpful for exploring the mechanism of gene expression in chondrocyterestrictive.

Genetic Variation in CAPN3 Gene and Its Relationship with Carcass Traits in Cattle

HOU Guanyu;ZENG Hongpu;WANG Dongjin;GUANSong;HUANG Xianzhou;XU Shangzhong

2010, 41(4): 398-402. doi:

Abstract ( 1166 )

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The 5′UTR of CAPN3 (Calpain3) gene was detected for searching SNPs and analyzing the distribution rule of different genotypes in different breeds, in order to study the relationship between CAPN3 gene and meat quality, meanwhile, searching the markers correlated with meat quality. SNPs were screened by sequencing and different genotypes were determined by polymerase chain reaction restriction fragment length polymorphism (PCRRFLP), then allele frequencies and polymorphism information content (PIC) were calculated. The result indicated that a new SNP (C→T) was found at 2 546 bp. The analysis result of the relationship between genotypes and productive performances by the Least Square Method (LSM) showed that the difference between AB and BB genotypes in carcass weight was significant (P<0.05). CAPN3 gene was primarily deduced to be a potential major gene or linked to major gene effecting meat quality traits. And this SNP might be a potential molecular marker of MAS.

Apoptosis of Tetraploid Blastocysts and Expression of Apoptosis Genes in Mouse

YUN Xinxu;FENG Shutang;MU Yulian;LI Kui

2010, 41(4): 403-409. doi:

Abstract ( 690 )

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The purpose of this study was to investigate the developmental potential of the mouse tetraploid embryos by the detection of the cell apoptosis and the expression of Bax and Bcl2 genes in the tetraploid blastocysts.The results indicated that the rate of the blastocyst formation was no significant difference between in the tetraploid embryos and in the diploid embryos in vitro. The result of the differential staining showed that the cell numbers of ICM and TE, the ratio of ICM/Total cells in the tetraploid blastocysts were lower than that in the diploid blastocysts in vitro or in vivo. At the same time,the detection of the cell apoptosis by TUNEL suggested that the index of the cell apoptosis in the tetraploid blastocysts was higher than that in the diploid blastocysts in vitro or in vivo. In addition, the expression of Bax gene in the tetraploid blastocysts was higher than that in the diploid blastocysts in vitro or in vivo, whereas there was no significant different expression of Bcl2 gene among them. The study demonstrated that the quality of the mouse tetraploid blastocysts was much worse than that of the diploid blastocysts in vitro or in vivo, which may cause the abnormal development of the mouse tetraploid embryos.

Effect of TSA on Fibroblast Cell as Donor Cell in the Tan Sheep

LI Xiangchen;YAO Yaxin;LIANG Suli;WANG Yipeng;GUAN Weijun;MA Yuehui

2010, 41(4): 410-416. doi:

Abstract ( 703 )

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In order to investigate the potential of fibroblasts as donor cells, and find out the optimal concentration of TSA used to treat fibroblasts, and further to improve the developmental level of cloned embryos, the fibroblasts were treated with 100, 50, 25, 10 ng·mL-1 TSA, cell morphology and viability were investigated. Flow cytometry were adopted to analyse cell cycle. Acetylation level was detected by indirect immunofluorescence, and the developmental level of cloned embryos was evaluated. Most cells treated with 100 ng·mL-1 TSA were dead, but a significant increase of acetylation level was detected; the cell cycle was arrested at G0/G1 checkpoint with the existence of TSA; generally，there was a higher proportion of cleavages and blastulas (P<0.05, (85.2±3.4)% vs( 68.6±6.7)%, (35.6±5.7)% vs (10.4±8.3)%) when the cells were treated with 10 ng·mL-1 TSA. The results suggested that acetylation level, cell morphology and embryonic developmental level were improved and optimal for the donor cells treated with 10 ng·mL-1 TSA.

Anatomical Changes of Digestive Tract and Rumen Functional Developmentin Grazing Lamb at the Age of 056 d

WANG Cailian;HAO Zhengli;LI Fadi;YU Yang;LANG Xia;MA Youji;NIAN Fang;GUO Jiangpeng

2010, 41(4): 417-424. doi:

Abstract ( 727 )

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To understand the anatomical development of whole digestive tract and functional development of the rumen in grazing lambs at the age of 056 d, fortyfour Gansu Merino male lambs were killed and sampled at 0, 3, 7, 14, 21, 28, 42, 56 days of age, respectively. The relative weights of gastric ventriculars(% live weight, RWL) of forestomach, abomasum, small intestine and cecum, relative weight(%of whole stomach weight, RWS) and volume(%of whole stomach volume, RCS) of stomach compartments, relative weight of the compartment contents(% of whole stomach contents weight, RWCS) and the pH of the compartment contents were measured; the concentrations of VFA and activity of microorganism enzyme in rumen content were also determined. With the age increasing, RWL of forestomach, small intestine and cecum increased with the faster increasing of forestomach and cecum at day 1421, followed day 4256, the increasing of small intestine slowly. RWS and RCS of rumen increased very markably, while these of abomasum reduced, all the changes were faster at day 714, 1421 and 4256. During 014 d, RCS was lower than RWS for forestomach, but thereafter the growing was faster for former than that for later, hence the value of RCS exceeded RWS after day 28. With the age increasing, the pH of contents in stomach compartments was stable relatively, the pH of rumen contents ranged from 6.03 to 6.67. 07 d the ratio of occurring rumen cellulase activities in partial detected lambs increased with the age increasing, after 14 d the enzyme activities had been observed in the all lambs tested, and the activities increased with the age increasing too; the peaks of activities occurred at day 14 for protease and at day 42 for amylase and cellulase. Acetate, propionate and butyrate were found from lambs′ rumen contents of day 3 and the concentration of acids raised with age increasing. The appearance of peaks of VFAs concentration and activities of the amylase and cellulase were parallel with the development of rumenreticulum. The faster growing during day 021 and thereafter the slower growing for RWS of forestomach was caused by the change of nutrient state of lambs. Under the study condition, lambs′ weaning is better at 2 monthold, and day 42 is the earliest weaning age accepted.

Effects of Intraduodenal Infusion of Soybean Small Peptide on Absorption of Small Peptide and Free Amino Acids in Dairy Goats

WANG Ling;;LIU Hui;LI Shengli;ZHAO Shengjun;WANG Libin;YANG Zaijun

2010, 41(4): 425-433. doi:

Abstract ( 690 )

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This experiment was conducted to study the effects of intraduodenally infusion of soybean small peptide (SSP) on absorption of small peptide and free amino acids in dairy goats. Seven dairy goats ((37.88±3.03) kg) were used in a 4×4 Latin square design. Goats were fitted with duodenal cannulas and implanted with indwelling catheters in carotid artery, mesenteric vein and portal vein. Each goat received four treatments respectively: infusion of normal saline, 60, 120 and 180 g·d-1 SSP into duodenum. Increased SSP duodenal infusion elevated the net fluxes of peptide bound amino acids (PAA) across mesentericdrained viscera (MDV) (P<0.05), but decreased the net appearance rates of PAA in mesenteric vein blood flow. SSP infusion increased net fluxed of PAA across portaldrained viscera (PDV) compared with that of the control (P<0.05), but there was no significant difference among those of three SSP infusion treatments（P>0.05）. SSP infusion did not change concentrations of glucose, insulin, growth hormone and IGF1 (P>0.05), but increased the concentration of urea nitrogen (P<0.05) in the plasma of jugular vein. The results indicated that SSP infusion increased the net fluxes of PAA and free amino acid (FAA) across the MDV and PDV, but decreased the net appearances of PAA in the MDV, which may be a result of decreased absorption, increased degradation of PAA in the absorptive cell, or both.

Construction and Screening of the Recombinant Attenuated CPV Expressing P12A3C Gene of FMDV Serotype O

WANG Jianke;ZHANG Yunde;ZHANG Qiang;WU Guohua;YAN Xinmin;ZHU Haixia;LI Jian;SHAO Changchun;ZHU Caizhu;WU Lei

2010, 41(4): 434-441. doi:

Abstract ( 815 )

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The constructed gene EGFPP7.5P12A3C of FMDV O/China99 strain was linked to the linear vector by blunt end ligation using KpnⅠ enzyme site, and the recombinant vector pUC119TKEGFPP7.5P12A3C was obtained. Homologous recombination between recombinant vector with deleted gene TK and capripoxvirus attenuated strain took place in the cell BHK21. The recombinant attenuated strain was screened by choosing EGFP as marker gene. And the recombinant virus was identified by PCR, antigen capture ELISA and Western blot analysis. The recombinant attenuated strain can passage steady in the first to the tenth generation of BHK21 cell, a fragment of about 3 000 bp was amplified，and the gene was confirmed as P12A3C by sequencing. The results of antigen capture ELISA were all positive. Western blot analysis showed that the corresponding protein was expressed in transfer vector pUC119TK EGFPP7.5P12A3C infected GTPV AV41 BHK21 cells and it could be recognized by serotype O of FMDV serum. The result demonstrates that the recombinant attenuated CPV containing P12A3C gene of FMDV serotype O were obtained.

Genome Sequencing and Genetic Analysis of One H6N5 Subtype Avian Influenza Virus A/duck/Yangzhou/013/2008

GU Min;ZHAO Guo;SONG Qingqing;LIU Wenbo;PENG Daxin;LIU Xiufan

2010, 41(4): 441-448. doi:

Abstract ( 1044 )

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One H6N5 subtype avian influenza virus, A/duck/Yangzhou/013/2008 (Dk/YZ/013/08), was isolated from aquatic birds in some live bird market in East China during our epidemiological surveillance and was identified by HA/HI test and specific RTPCR test. To investigate the biological role of H6N5 avian influenza viruses in the influenza ecology, Dk/YZ/013/08 was subjected to genome sequencing. All the complete genome sequences of H6N5 subtype influenza viruses available in GenBank and some other reference sequences were retrieved to do the phylogenetic analysis with Dk/YZ/013/08. The results indicated that the HA gene showed 94% nucleotide identity with A/duck/Kingmen/E322/2004(H6N2) isolated from Taiwan Province of China, and the amino acid motif of cleavage site between HA1 and HA2 of Dk/YZ/013/08 was “PQIETRG”, which was the typical characterization of the LPAIV. The NA gene of Dk/YZ/013/08 had close relationship with A/mallard/Switzerland/WV4060167/2006(H3N5). However, the PB2 gene displayed 97.3% similarity with A/duck/Zhejiang/11/2000(H5N1) and clustered with A/goose/Guangdong/1/1996(H5N1). Our results indicated that Dk/YZ/013/08 may be a reassortant virus by introducing its PB2 gene from H5N1 subtype influenza virus.

Evaluation of Three Serological Tests for Brucellosis in Cattle Using Bayesian Analysis"

LIU Xiaomin;YANG Hua;GAO Mingchun;LI Yanwei;WANG Junwei

2010, 41(4): 449-453. doi:

Abstract ( 734 )

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The objective of our study was to evaluate the diagnostic performance of three serological tests for brucellosis. Serological test results were used to compare the performance of Rose Bengal Test (RBT), indirect ELISA (IELISA) and Fluorescence Polarisation Assay (FPA) in diagnosing brucellosis in cattle. Their performance was evaluated using Bayesian analysis which allows the sensitivity (Se) and specificity (Sp) to be estimated in the absence of a gold standard. The sensitivity of RBT, FPA and IELISA were 91.81%, 95.92% and 95.98%, respectively; and specificity of RBT, FPA and IELISA were 98.80%, 86.54% and 98.65%, respectively. The results demonstrated that the IELISA performing well in diagnosing brucellosis in cattle.

Prediction on the Status of HPAI and Its Risk Trend in China

ZHANG Zhicheng;LI Changyou;HUANG Baoxu;LIU Yongjun;SONG Jiande;WEI Xinjie;CAI Lijuan;WANG Zhiliang;MA Hongchao

2010, 41(4): 454-462. doi:

Abstract ( 1197 )

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The objective of this study was to explore the status of poultry populations’ highly pathogenic avian influenza and its range developing risk. Integrated with the data mining techniques and theory of risk assessment of OIE (Organisation Mondiale de la Santé Animale),combined with the application of the methods of ecological pattern analysis and Bernoulli statistical model, the risk status of highly pathogenic avian influenza and its range expansion trend were derived since its first reported in 2004 in China. Results of analysis showed:(1)China has a very big host populations of poultry in certain given geographical regions, characterized by its complex raising & feeding pattern & mode, and diversified varieties, which make the consecutive occurrences of HPAI possible, and thereof entailed the bigger challenges for HPAI risk control. (2)The occurrences of HPAI in China featured by its significant tempospatial patterns, mainly characterized by its seasonalregional and specified host population occurrences, which indicted the importance of scientifically regional risk management and regional cooperation. (3)In risk developing trend, due to the regionalnational and international occurrences of HPAI, and existence of larger exposure host populations, make the further risk of HPAI infection and expansion possible in given region. (4)In risk predictions, Bernoulli statistical model integrated with geographical informational techniques were applied, result showed that there may be 210 times of HPAI occurrences with 90% confidence interval (CI) in 2008,6 times of HPAI occurrences were most likely, and poultry populations in southern part of China may be exposed at possible risk. There exist a highly likelihood for the reoccurrences of HPAI among poultry populations in China in future.

Sequence and Phylogenetic Analysis of Mitochondrial cox1 and nad1 Genes for Spirometra erinaceieuropaei Isolates from Hunan Province

LIU Zikui;LIU Guohua;DAI Rongsi;LIU Wei;LI Fen;HU Tao;LIU Yi

2010, 41(4): 463-468. doi:

Abstract ( 1024 )

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The objectives of the present study were to examine sequence variation in the mitochondrial cytochrome coxidase subunit 1 (cox1) gene and NADH dehydrogenase subunit 1(nad1) gene among Spirometra erinaceieuropaei isolates from Hunan Province，and to reconstruct their phylogenetic relationship using cox1 and nad1 sequences. The partial cox1 (pcox1) and nad1 (pnad1) was amplified from each S. erinaceieuropaei sample，and pcox1 and pnad1 sequences were aligned using the ClustalX 1.81. MP and NJ trees of pcox1 and pnad1 were constructed using the software Phylip 3.67 version 4.0 and Mage version 4.0, and ML tree was also constructed using Puzzle version 5.2. Sequence homology analysis was performed using the Megalign program of the software DNAStar version 5.0. The results showed that the lengths of pcox1 and pnad1 sequences were 396 and 566 bp, respectively. The constructed phylogenetic tree revealed that the Hunan isolates and the S. erinaceieuropaei available in GenBank were clustered in the same clade. There is no significant variation in pcox1 sequences within S. erinaceieuropaei, while interspecies difference is obvious. It is concluded that pcox1 and pnad1 sequences can be used as genetic marker for population genetic studies of cestodes. The results of the present study provided foundation for further studies of population genetics of S. erinaceieuropaei，and for diagnosis of the resultant disease.

Cloning and Functional Analysis of Cyclin A Gene in Pig

FAN Li;TANG Qinghai;ZHANG Yanming;LIU Wei;TONG Gang

2010, 41(4): 469-477. doi:

Abstract ( 673 )

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The study was aimed at cloning porcine Cyclin A gene and expressing it in swine umbilicus veins endothelial cell (SUVEC) to verify its functionality. The human Cyclin A gene was utilized as the informational probe to eclone the pigs Cyclin A gene. Reverse transcription polymerase chain reaction (RTPCR) was used to identify the products in silico cloned, and the bioinformatics methods was used to analyze structure characteristics of the gene, RTPCR, Western blot were employed for investigating its expression in SUVEC. Subcellular localization of Cyclin A was analyzed by confocal microscope, and the flow cytometry was used to analyze the cell cycle, and MTS was used to detect the proliferation of the cells. The results proved that the product of RTPCR was consistent with that of silico cloning. This cDNA contains the complete open reading frame (ORF) of 1 299 bp coding 432 amino acid residues，and NCBI BLAST analysis indicated that the gene is located in swine on chromosome 8. Western blot manifested that molecular size of Cyclin A protein was about 40 kDa, and subcellular localization showed that this protein was localized in nuclear. The flow cytometry analysis showed that cell lines SUVECCycAGFP of G1 phase cells increased by 15% to 20%, while those in S phase decreased about 18% compared with that of the control cells. MTS assay showed that the proliferative activity of cell lines SUVECCycAGFP was significantly higher than that of the control groups. Our results indicate that Cyclin A gene of pig was successfully cloned and its biological function was also confirmed, which will provide a foundation for further research on the impact of virus infection on the Cyclin A.

Expression of Cyclin Dependent Kinase 5 in Alpaca Skin of Different Hair Colors

LIU Jia;DONG Chang-sheng;FAN Rui-wen;HE Jun-ping;GAO Lei;DONG Chun-guang;DONG Yan-jun

2010, 41(4): 478-483. doi:

Abstract ( 685 )

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This experiment was conducted to explore the expression and function of cyclin dependent kinase 5 in alpaca skin. The whole testes were obtained from adult alpacas of white and brown hair colors. The localization of CDK5 in alpaca skin was examined by immunohistochemistry, and comparative mRNA expression quantity of CDK5 in alpaca skin of different coat colors was analyzed by QRTPCR. The results showed that the positive signals were found in hair bulb and outer rootsheath of hair follicles in alpacas, the difference of expression was significant between the white and brown alpaca′s based on the average optical density(P<0.05). The result of QRTPCR showed that the comparative mRNA expression quantity of CDK5 in brown alpacas was 1.624 5 times than white ones. The findings showed that CDK5 may involved in the regulation of hair color formation.

Effect of Tiletamine on the Endogenous Opioid Peptides Content in DifferentEncephalic Regions of Rats

ZHANG Yan;WANG Hong-bin;FAN Hong-gang

2010, 41(4): 484-488. doi:

Abstract ( 1088 )

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To explain the possible molecular mechanisms of tiletamine anesthesia on the central nervous system by investigating the changes of endogenous opioid peptides content in different encephalic regions in rats. 8 Wista rats were allocated randomly from 128 Wista rats as control group. The rest rats were divided randomly into L-Enk, M-Enk, β-EP, dynA and OFQ groups，and each group of animals were divided randomly into maintenance of anesthesia group，recovery from anesthesiaⅠgroup and recovery from anesthesiaⅡgroup. The content of 5 kinds of endogenous opioid peptides in different encephalic regions in rats were measured by Enzyme-Linked Immunosorbent Assays(ELISA). The L-Enk, M-Enk, β-EP and dynA contents in the cerebral cortex and the thalamus in the anesthesia group were obviously increased as compared with control group(P<0.05 or P<0.01)，while the OFQ content was significantly decreased(P<0.05 or P<0.01).The L-Enk, M-Enk and β-EP contents in the hippocampus were obviously increased as compared with control group(P<0.05 or P<0.01)，but neither the dynA nor OFQ contents were obviously changed. The M-Enk, β-EP and dynA contents in the brainstem were obviously increased as compared with control group(P<0.05 or P<0.01)，the OFQ content was obviously decreased（P<0.05），but the L-Enk content was not changed. In both of the recovery from anesthesiaⅠgroup and anesthesiaⅡgroup the L-Enk, M-Enk, β-EP, dynA and OFQ contents in the above encephalic regions were revived as compared with control group(P>0.05).The content of 5 kinds of endogenous opioid peptides in the cerebellum were not obviously changed in the process of general anesthesia of tiletamine. The results suggest that tiletamine anesthesia have an effect on the content of 5 kinds of endogenous opioid peptides in different encephalic regions. Tiletamine could increase the L-Enk , M-Enk, β-EP and dynA contents in both of the cerebral cortex and the thalamus，and increase the L-Enk, M-Enk and β-EP contents in the hippocampus，and M-Enk, β-EP and dynA contents in the brainstem，respectively，but decrease the OFQ content in the cerebral cortex，the thalamus and the brain stem.

Inhibition Activity of Coptidis rhizoma Dicoction against Riemerella anatipestifer and Its Effects on Morphology of Bacterial Body

ZHANG Xian-fu;WEI Qiang;BAO Guo-lian;JI Quan-an;PANG An-na

2010, 41(4): 489-494. doi:

Abstract ( 640 )

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To investigate inhibition activity of Coptidis rhizoma dicoction against Riemerella anatipestifer and its effects on morphology of bacterial body, Coptidis rhizoma dicoction were prepared with water extraction and alcohol sedimentation method. After vacuum distillation, Ethanol extracts of Coptidis rhizoma decoction (EeCrd) which contained crude drug 176 g·L^-1 was acquired. This extracts was bi-diluted to make graded concentrations which contained crude drugs 88,88×2^-1，88×2^-2 ……88×2^-9 g·L^-1, Minimum inhibitory concentration (MIC) and sub-inhibited concentration of EeCrd on Riemeralla anatipestifer in vitro were determined by both disc diffusion test and tube broth assays. Morphology of RA were observed by transmission electron microscope in concentration groups of 88 and 88×2^-3 g·L^-1 of EeCrd in 17 and 3, 6, 17 h, respectively. MIC of EeCrd on RA is 88×2^-2 g·L^-1 concentration group by both two methods, sub-inhibited concentration are 88×2^-3 g·L^-1. Above 88×2^-4 g·L^-1 concentrations, EeCrd almost have no any antibacterial activities. Under 88 g·L^-1 concentrations of EeCrd with 50 μL in 17 h, lengths of RA body concentrated and got shorter, ultimately died. In 88×2^-3 g·L^-1 of EeCrd in 3, 6 , 17 h, lengths of RA body became longer, cell wall got thinner, the fold inside the body reduced, the texture becomes uneven, finally withered, folded, bent and died. EeCrd achieve the purpose of inhibiting the proliferation of RA by destroying the structure of RA cell wall.

The Observation of the Apoptosis Induced by Muscovy Duck Reovirus

QI Bao-min;CHEN Xiao-yan;WU Bao-cheng;YAO Jin-shui;ZHANG Hong-lei

2010, 41(4): 495-499. doi:

Abstract ( 652 )

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10-day old Muscovy ducks were infected with Muscovy Duck Reovirus. Apoptosis were detected and the expression of FasL were studied in many infected tissues post infection, using in-situ end labeling technique and immunohisto chemistry method. The result of in-situ end labeling revealed that a large number of positive cells were found in many tissues， which showed that Muscovy duck reovirus could induce varying-degreed apoptosis in the liver，spleen，kidney，lung，thymus，cecum and bursa of fabricius. The result of immunohistochemical study showed that the expression of FasL were observed in liver, kidney, lung，thymus，cecum and bursa of fabricius，and the changes of FasL expression were highly correlated with the level of apoptotic index. The results showed that the mechanism of apoptosis induced by Muscovy duck reovirus is closely related with the expression of FasL.

The Role of Mitochondria Mediated Apoptosis Pathway in Sertoli-germ CellApoptosis Induced by Manganese in Cocks

BI Ming-yu;LI Jin-long;LI Shu;CHEN Lei;ZHANG Zi-wei;TANG Hong-peng;XU Shi-wen

2010, 41(4): 500-504. doi:

Abstract ( 712 )

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The aim of this study was to discuss the role of mitochondria mediated apoptosis pathway in Sertoli-germ cell apoptosis induced by manganese in cocks. Cock Sertoli-germ cells were cultivated in the DMEM for 24 h added with MnCl2, whose final concentrations were 0, 2, 3, and 4 mmol·L^-1 respectively. The apoptosis was detected by AO/EB double staining. Mitochondrial transmembrane potential was detected by flow cytometer. Expression of Cytochrome c (Cytc) in cytolymph was detected by western blotting. The activities of Caspase-9, 3 were examined by spectrophotography. Compared with control group, the apoptosis index (AI) of cock Sertoli-germ cells was significantly increased (P＜0.01), mitochondrial transmembrane potential was significantly decreased (P＜0.01), expression of Cytc in cytolymph was increased, and the activity of Caspase-9, 3 were increased (P＜0.01) in 2, 3, and 4 mmol·L^-1 MnCl₂ group. The apoptosis mediated by mitochondria pathway was one of the reproductive toxic mechanisms caused by manganese.

Migration and Accumulation of Primordial Germ Cells in the Quail Earlier Embryo

CHANG Guo-bin;CHENG Xu-mei;LI Bi-chun;LIU Xiang-ping;QIN Yu-rong;CHEN Rong;CHEN Guo-hong

2010, 41(4): 505-510. doi:

Abstract ( 1010 )

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This paper was conducted to study the migration and accumulation of primordial germ cells (PGCs) in the quail earlier embryo, aiming to provide the base of transgenic technique. The distribution and amount of PGCs at different stages were identified specially by the monoclonal antibody, QH1. The results showed that quail PGCs originated from the area opaca of unincubated blastoderm, and then transferred to area pellucida and the germinal crescent area. About 27 h post-incubation, a few PGCs first appeared in blood vessels of the area pellucida, where a great lot of PGCs accumulated at 36 h post-incubation. Subsequently, the single and mass PGCs were found from the head to omphalo mesenteric and mainly settled down the mesenchymal blood vessels of head. The PGCs amount at different stages were different, and had two periods of maximum in whole stages, namely, primitive streak(6 h post-incubation) and tenth somite (36 h post-incubation). The results indicated that QH1 could identify the PGCs in the quail earlier unincubated embryo, where the PGCs distributed randomly, and there founded two periods of maximum proliferation in whole stages.

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