Acta Veterinaria et Zootechnica Sinica

ZHANG Chenfei;CHEN Changhai;LIU Yaoxing

2012, 43(5): 669-675. doi:

Abstract ( 360 )

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Nipah virus is a newly identified paramyxovirus that causes serious respiratory disease and encephalitis in humans and animals. Outbreaks and spreads of this virus have led to the severe economic losses and human deaths since the first isolation of Nipah virus in Malaysia, 1999. Bats in the genus Pteropus are natural hosts of this viruses, and contact with infected animals or humantohuman transmission were thought to be the major route of the virus spreading. As is highly contagious in human and swine with high case fatality rate, Nipah virus are classified as biosafety level 4 (BSL4) agents, for which there is still no available vaccination or effective antiviral treatment. This review provides a simple overview of Nipah virus in taxonomy, genomics and proteomics, epidemiology, vaccine development, clinical and pathological signs, diagnostic techniques.

Research on Polymorphism of Hal, RN and FTO Genes Related to Meat Quality in Different Pig Breeds

TAO Xin;DENG Bo;MEN Xiaoming;XU Ziwei

2012, 43(5): 676-683. doi:

Abstract ( 451 )

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To investigate the genetic variability of Hal, RN and FTO genes related to meat quality among foreign and Chinese pig breeds, the polymorphism of these genes in four pig breeds including Duroc, Landrace, Large Yorkshire and Jinhua pig were detected by PCRRFLP, respectively. The genetic variation of g.276G>T and c.594C>G in FTO gene were analyzed. The results showed as follows: (1) For Hal gene, HalNHaln genotype was detected and its frequency was 0.166, but no HalnHaln genotype was detected in Duroc pig. Only HalNHalN genotype was found in Landrace, Large Yorkshire and Jinhua pig. (2) For RN gene, only rn/rn genotype was detected in four pig breeds. (3) At g.276G>T site in FTO gene, three breeds including Duroc, Landrace and Large Yorkshire pig had polymorphism, but Jinhua pig had no polymorphism. At c.594C>G site in FTO gene, all the breeds had polymorphism. However, higher CC genotype frequency was found in Duroc, Landrace and Large Yorkshire pig, but in Jinhua pig GG genotype frequency was higher. These results indicate that higher frequency of HalNHaln genotype has existed in some pig breeds because Hal gene show benefits and defects for pig production. Moreover, FTO gene could be regarded as a candidate gene affecting meat quality in special pig populations according to its known functions and the previous reports. This result show that the genotypes of FTO are very significantly different between the foreign breeds and Chinese indigenous breeds, which provide some data for further study on the molecular mechanism of meat quality of different pig breeds.

Gene Clone, Subcellular Localization of Expression Products of HFABPand the Preparation of Transgenic Mice in Xuhuai Goat

YIN Yanhui;WEI Guanghui;LI Wei;ZHU Caiye;ZHANG Yani;DU Lixin;CAO Wenguang;LI Bichun

2012, 43(5): 684-692. doi:

Abstract ( 381 )

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The purpose of this study was to clone hearttype fatty acid binding protein (HFABP) gene cDNA of Xuhuai goat, and to explore its bioinformatics function and the possibility of preparation of transgenic animals among heterogeneous species. The subcelluar location HFABP was detected by EGFP fusion protein and its expression was observed in vitro. Reverse transcription PCR (RTPCR) technology was used to clone the HFABP gene cDNA of Xuhuai goat, its biological information characteristics was analyzed by online software, then the expression vector pEGFPHFABP was constructed. The transfection of goat fibroblasts (GEF) was performed by Liposomes (LTX), and fluorescence was observed under inverted microscope after 48 h. The RTPCR was conducted to detect mRNA expression of HFABP in GEF. The pEGFPHFABP was injected into mouse testicular and its expression was detected at the level of DNA and protein. The complete CDS size of HFABP was 402 bp, encoding 133 amino acids with GenBank accession number (AY466498.1). The HFABP cDNA coding sequence was compared with the corresponding regions of human, chicken, brown rat, cow, wild boar, donkey and zebra fish, the similarity was 89%, 76%, 85%, 84%, 93%, 91%, 70%, respectively, amino acid sequence homology was 90%, 79%, 88%, 97%, 95%, 94%, 72%, respectively. The signal peptide was not found in HFABP protein. The RTPCR results showed the HFABP mRNA expressed successfully in vitro. pEGFPHFABP was successfully constructed, and HFABP mRNA was expressed. The HFABP protein was localized in the cytoplasm which was in line with the result of online prediction. The gene can aslo be expressed in mice transiently and persistencely after intravenous and testicular injection. The HFABP gene cDNA of Xuhuai goat was cloned successfully, and it was conservative during the evolutionary process, there was no signal peptide in protein. The HFABP protein was located in the cytoplasm, and also could be expressed in mice successfully.

Bioinformatics Analyzing of the Differential Expression of Genes Affecting the Prolificacy in Ovarian Tissue in Goat

FENG Anxue;QIAN Lili;JIA Qing;XING Zengxi

2012, 43(5): 693-700. doi:

Abstract ( 386 )

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The purpose of this study was to investigate the hereditary basis of high prolificacy in goat, the DDRTPCR was applied to screen 4 gene fragments differentially expressed as candidate genes affecting the reproductive traits in the ovarian tissues of local native goat of the midst of Hebei with different fecundity levels during oestrum. According to the further analysis result of homology and transcription factor binding sites, two segments were newly found. One had 81% homology with cloned fragment (THY010003E05) of wild boar, but the function was unknown. Another segment was shared 94% homology with the pcnp mRNA gene in cattle. Accordingly, the pcnp gene of goat was amplified, cloned and sequenced. By using bioinformatics softwares and methods, the chemicophysical properities, secondary structure, signal peptide, nucear localization sequence, phosphorylation sites, transmembrane region and protein functional domains of the amino acid sequences coded by pcnp gene were predicted. Compaerd with complete CDS sequence of pcnp gene of cattle,the sequence homologous of pcnp in goat was 100%, coding region was 537 bp encoding 179 amino acids. It was suggested that PCNP was a hydrophilic nucleoprotein.Random coils and αhelix were the main structural type of the flexible region in secondary structure, and there were partial extended chains and βturn. 5 proteins interacting with PCNP protein were predicted, and 3 low complexity regions were discovered. It was suggested that PCNP was relatively conservative, it was most likely to play a role in the nucleus and affect goat prolificacy via involving in cell cycle regulation or gene expression.

Genetic Variation and Phylogeney of Jianghuai Buffalo Based on MitochondrialDNA Dloop and Cytb Gene

LU Yongfang;CAI Zhihua;LI Qiang;ZHANG Yi;ZHANG Qin;ZHANG Yuan

2012, 43(5): 701-707. doi:

Abstract ( 445 )

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To study the molecular germplasm characteristics of Jianghuai buffalo, the genetic diversity and phylogeny of mitochondrial DNA Dloop region and the complete sequence of cytochrome (Cyt b) gene of 82 Jianghuai buffaloes sampled from two subpopulations were investigated. These data were analyzed together with the published Dloop sequences of 141 Chinese buffaloes in GenBank. A total of 104 haplotypes and 91 nucleotide polymorphic sites were identified. Thirtytwo Dloop haplotypes were newly found only in Jianghui buffalo. The estimated overall nucleotide diversity and haplotype diversity were (0.015 43±0.001 42) and (0.948±0.009), respectively. The genetic variability of Jianghuai buffalo was similar with those of other Chinese buffalo populations, with nucleotide diversity to be (0.014 89±0.002 32) and haplotype diversity to be (0.955±0.013), which indicated the rich mitochondrial genetic diversity in this population. Phylogenetic tree and network analysis revealed two mtDNA lineages, the A and B lines of swamp buffaloes, and the B line could be further divided into sublineages of b1 and b2, which indicated that at least there were two maternal swamp buffalo origins in Jianghuai swamp buffalo population. The results based on the phylogenetic analysis of Cytb gene also supported this conclusion observaed in the Dloop region of Jianghuai buffalo. These findings lay a foundation for conservation and utilization of Jianghuai buffalo genetic resources in the future.

Associations of UCP2 Gene Polymorphisms with Growth Traits in Simmental

CHEN Cui;ZHANG Limin;CHEN Xiaojie;LIU Xidong;ZHANG Meng;LI Jiao;YUAN Zhengrong;ZHANG Lupei;GAO Xue;GAO Huijiang;LI Junya;XU Shangzhong

2012, 43(5): 708-716. doi:

Abstract ( 346 )

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The aim of this study was to analyze the association of UCP2 gene polymorphisms and haplotypes with feed conversion ratio(FCR) and growth traits in beef cattle. In this study, 118 Simmental were used, four SNPs of UCP2 gene (G118A，C161T，C215G，C305T) were genetyped by PCRRFLP in Simmental to investigate the associations of SNPs and haplotypes with FCR and growth traits. The results showed that G118A, C161T and C305T were significantly associated with FCR (P<0.05).Compared with the individuals with homozygote, the values of FCR of individuals with the heterozygote were significantly higher. However, the difference between the individuals with homozygotes were not significant. The locus C161T and C305T had an major affect on heart girth and circumference of abdomen. Among the phenotypes which were significantly different, individuals with TT genotype had a significantly higher average value than that of individuals with AA genotype. Association analysis of haplotypes indicated similar results with that of SNPs. Considered the FCR,ADG and body size traits, it was suggested that the TT genotype at locus C161T and C305T of UCP2 gene were important candidate molecular markers for improving growth traits.

The Role of Malic Enzyme in Goose Hepatic Steatosis Induced by Glucose and Insulin

XIA Lu;PAN Zhixiong;TANG Hui;XIANG Shuxia;WANG Jing;HAN Chunchun;LI Liang;WANG Jiwen

2012, 43(5): 717-722. doi:

Abstract ( 348 )

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The experiment was conducted to study the enzyme activity and mRNA expression of malic enzyme (ME) induced by glucose and insulin in goose primary hepatocyte, and identify the role of malic enzyme in goose hepatic steatosis. In this study, partial sequence of ME gene was cloned using Sichuan White goose. The goose primary hepatocyte was cultured and hepatic steatosis was induced by means of adding different concentrations of glucose and insulin, malic enzyme activity and mRNA expression and the triglyeride (TG) content in cells tested were detected. The goose ME gene sequence had highest similarity with Gallus gallus. Compared with the control group, glucose and its synergy with insulin could significantly increase the TG content in a dose dependent manner, however, insulin alone failed to notably affect TG content; the enzyme activity and mRNA expression of ME gradually increased with glucose concentrations increasing, and 30 mmol·L-1 glucose could significantly increase those parameters（P＜0.05）; low contents of insulin（50 nmol·L-1）could largely elevate the enzyme activity and mRNA expression of ME(P＜0.05), which could be suppressed by 200 nmol·L-1 insulin. In addition, the synergy of glucose with lower content of insulin（50 nmol·L-1） could significantly increase the enzyme activity and gene expression of ME. In this study, the partial sequence of goose ME gene were cloned. Meanwhile, glucose and insulin could coordinately promote the mRNA expression and enzyme activity of ME in goose liver resulting in TG accumulation.

Genetic Diversity of IL10 Gene Exons in 5 Rabbit Populations

MAO Liuliu;PAN Yulai;WANG Xiaoming;WAN Xiaoying;LI Bichun;WU Xinsheng;

2012, 43(5): 723-728. doi:

Abstract ( 364 )

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This experiment was conducted to study the polymorphism of IL10 gene exons in rabbit and aimed to provide a theoretical foundation for further reseach on correlation between rabbit IL10 gene and disease resistance. Five exons of IL10 gene in 5 rabbit populations including Beaver Color Rex rabbit,White Rex rabbit,Wanline Angora rabbit,Jiuyishan rabbit and Minxinan Black rabbit were scanned by PCRSSCP method with five specific primers designed according to the IL10 sequence from GenBank. The results showed that 10 genotypes and 4 alleles were found in exon3 of IL10 gene which contained three SNPs, 3 genotypes and 2 alleles were found in exon4 which contained one SNP. There was no mutation found at exon1, exon2, exon5 of IL10 gene. In exon3, D allele was found only in Minxinan Black rabbit and Jiuyishan rabbit.The distribution of these genotypes in all populations was consistent with the HardyWeinberg equilibrium except Beaver Color Rex rabbit and Minxinan Black rabbit, which was very significantly different (P<0.01) among different populations. In exon4, A1B1 genotype was not found in Rex rabbit group.The distribution of these genotypes was significant different (P<0.05 or P<0.01) among different populations except between Beaver Color and White Rex rabbit (P>0.05). These results indicated that genetic diversity of rabbit IL10 gene was detected in exon3 and exon4.

Expression and Location Analysis of PAX3 Transcription Factor in Alpaca Skin

ZHU Zhiwei;HE Junping;YU Xiuju;LI Pengfei;CHENG Zhixue;DONG Changsheng

2012, 43(5): 729-734. doi:

Abstract ( 332 )

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This experiment was conducted to study PAX3 expression and location in alpaca skin. In the study, the distribution of PAX3 protein was studied by immunohistochemistry，the expression level was analyzed by ImagePro Plus software and Western blot. The result showed PAX3 protein was mainly located in melancytes during the basal cells of hair bulb, around dermis papilla and in hair root. By ImagePro Plus software and Western blot analysis, the protein expression level was significantly different in different coat color (P<0.05). Based on the result, it indicates that PAX3 plays an important role in the melanocytes proliferation, differentiation and pigmentation.

In vitro Marker of Soybean Agglutinin Binding Sites in Porcine Intestinal Epithelial Cells

CHE Dongsheng;MU Chenglong;LIU Feifei;ZHANG Haiquan;SUN Zewei;QIN Guixin;

2012, 43(5): 735-739. doi:

Abstract ( 356 )

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This study aimed to culture in vitro and separate piglet small intestines epithelial cells and observe the soybean agglutinin binding sites. Newborn and health piglets with nonlactation were selected, and the intestinal epithelium cells were cultured and purified. The intestinal epithelium cells purified were identified by immunochemistry method taking the cell keratin 8 antibodies as the first antibody. The intestinal epithelium cells identified were used to mark the soybean agglutinin binding sites with FITCSBA probe by cell agglutinin fluorescent chemistry marker test. The results showed that the porcine small intestinal epithelium cells purified were positive cells by cell keratin 8 antibodies identification, the purity was more than 90%, and the pig small intestine epithelial cells were positively marked by FITCSBA. The porcine small intestinal epithelium cells cultured in vitro posses the soybean agglutinin binding sites, and further specify the porcine small intestinal epithelium cells is one of the main cells combined by soybean agglutinin, which provide the theory for the research on monogastric animal soybean agglutinin antinutrition mechanism.

Effect of Zinc Lactate on Cell Proliferation and Relatedregulatory Genes mRNA Expression in Porcine Jejunal Epithelial Cells IPECJ2

HAN Guoquan;YU Bing;CHEN Daiwen;XIANG Zhentian;QI Hongwei;CHEN Hong;MAO Qian;MAO Xiangbing;HUANG Zhiqing

2012, 43(5): 740-747. doi:

Abstract ( 507 )

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The present study was conducted to evaluate the effects of zinc lactate on cell proliferation of jejunal epithelial cells IPECJ2 and mRNA expression of relatedregulatory genes in porcine. IPECJ2 cells were cultured with mediums added with zinc lactate by the zinc concentrations of 50, 100, 150 and 200 mg·L-1, respectively. Cell proliferation was estimated by Colorimetric analysis. Simultaneously, realtime quantitative RTPCR was applied to detect the mRNA expression of relatedregulatory genes (ZnT2, DMT1, IREG1, MT1 and ZIP4), and TBP mRNA level was used as the control. During the first 36 hours after treatment with zinc lactate, there was almost no effect on IPECJ2 cell proliferation. Subsequently, with increasing of the concentration of zinc, the rate of cell proliferation gradually increased. At the same time, after IPECJ2 cell was treated with zinc lactate, the expression of ZnT2, DMT1, IREG1and MT1 mRNA was increased with the increase of the concentration of zinc while the expression of ZIP4 mRNA decreased. Zinc lactate could promote cell proliferation of IPECJ2, upregulated the mRNA expression of ZnT2, DMT1, IREG1, MT1 and downregulated the mRNA expression of ZIP4.

Effects of Chronic Heat Exposure on Growth Performance,Meat Quality and Fat Deposition in Male and Female BeijingYou Chickens

HAO Jingyu;LU Qingping;ZHANG Hongfu;ZHANG Xiaodi

2012, 43(5): 748-754. doi:

Abstract ( 392 )

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The current study was conducted to investigate the effects of chronic heat exposure on growth performance, meat quality and fat deposition in BeijingYou (BJY) chickens. One hundred and fortyfour 49dayold healthy BJY chickens (72 males and 72 females) with similar body weight (male (608.58±13.07)g, female (518.50±8.33)g) were randomly assigned into two treatments(hightemperature group(35±0.37)℃,optimaltemperature group(25±0.38)℃) with 6 replicates in each treatment and 6 birds in each replicate. The experiment lasted for 23 d. The growth performance (111,1222 and 122 d) and the carcass traits, meat quality and fat deposition on 12 and 23 d of BJY chickens were assayed. The result showed as followed；1) During 111 d, male and female weight gain was significantly decreased by heat exposure (P<0.01), but the ratio of feed to gain was not affected (P>0.05). During 1222 d, heat exposure depressed weight gain of male (P<0.01), and ratio of feed to gain of males increased by heat exposure (P<0.01); 2) Meat quality of males and females were not affected by heat treatment (P>0.05); 3) For males, abdominal, intramuscular and subcutaneous fat deposition, carcass proportion, and leg weight proportion were not affected by heat exposure on 12 d (P>0.05). Whereas, on 23 d, abdominal, intramuscular and subcutaneous fat deposition markedly decreased by heat exposure (P<0.01), but carcass proportion and leg weight proportion increased (P<0.05). For females, abdominal and intramuscular fat deposition decreased by heat exposure on 12 d (P<0.05). On 23 d, heat exposure decreased subcutaneous fat deposition (P<0.05), and abdominal and intramuscular fat deposition, carcass proportion, and leg weight proportion were not affected by heat treatment (P>0.05). These results suggest that 12 d heat exposure significantly depressed male and female weight gain. With the prolonged exposure time, weight gain of males decreased more severely, but females were not significantly affected, and fat deposition of males gradually decreased, but decreasing degree in females were relieved.

Effect of Somatic Cell Count and Low Lactose in Milk on Changes of Whey Proteome

SHEN Weijun;YANG Yongxin;WANG Jiaqi;YUAN Tingjie;BU Dengpan;YANG Jinhui;ZHOU Lingyun

2012, 43(5): 755-760. doi:

Abstract ( 372 )

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The aim of the current study was to investigate the changes of the whey protein when the somatic cell count increased and lactose content decreased in milk. Milk was classified into milk with low lactose and high somatic cell count, and control group according to concentration of lactose and somatic cell count in milk. Whey proteins were collected by ultracentrifugation and separated by twodimensional gel electrophoresis, stained with Coomassie Blue G250 solution, and identified by matrix assisted laser desorption ionization time of flight mass spectrometry. βcasein was decreased, while whey proteins including serum albumin, haptoglobin, lactoferrin, transferrin and cathelicidin 1 increased in milk with low lactose and high somatic cell count group. The results suggested that the increase of whey proteins in milk was associated with concentration of low lactose, which may be the defense response of mammary gland to the damage of integrity of mammary gland epithelial cells when lactose content decreased in milk.

Coadministration with Inactivated Avian Influenza Virus and Adjuvants Enhances the Antibody Secreting Cells in Duck Respiratory Tract

KANG Haihong;SHEN Yumeng;WANG Hongli;YANG Qian

2012, 43(5): 761-766. doi:

Abstract ( 328 )

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Ducks were intranasal immunized with inactivated avian influenza H9N2 virus (IAIV) in combination with sodium taurodeoxycholate hydrate and CpG DNA. Distribution and number of antibody secreting cells in duck respiratory tract were studied. Results showed that 3, 5 and 7 weeks after vaccination with sodium taurodeoxycholate hydrate and CpG DNA, the area of IgA and IgG secreting cells of nasal cavity, trachea and tracheal bifurcation were increased significantly （P<0.05 or P<0.01） compared with that of ducks with IAIV alone. After intranasal immunization with IAIV and CpG DNA, the area of IgA and IgG secreting cells of nasal cavity and trachea were partly increased (P<0.05). While no significant change on the area of IgA and IgG antibody secreting cells of respiratory tract tissues was observed in respiratory tract between ducks with IAIV alone and PBS. It demonstrated that intranasal immunization with IAIV combined with sodium taurodeoxycholate hydrate and CpG DNA could increase the area of IgA antibody secreting cells and IgG antibody secreting cells and enhance the level of local humoral immune response of duck’s respiratory tract.

Development and Application of TaqMan Fluorescent Realtime Quantitative PCR for the Detection of Egg Drop Syndrome Virus

MA Zhenyuan;LI Gang;LI Wenchao;GUO Yufei;

2012, 43(5): 767-772. doi:

Abstract ( 377 )

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To establish the fluorescent realtime quantitative PCR (FQRTPCR) assay for rapid detection of egg drop syndrome virus, a pair of primers and a TaqMan probe were designed according to the sequences from highly conserved regions of the hexon protein gene of egg drop syndrome virus (EDSV). A positive recombinant plasmid was employed as standard template for the construction of standard curve. The detection limit of the assay was 10 copies DNA·μL-1 and the FQRTPCR was reproducible, as shown by satisfying its wide dynamic range from 101 to 108 copies DNA·μL-1. This assay is specific and has no crossreaction with DNA of other avian virus. The quantity of EDSV in eggs after artificial infection with NE4 strain of EDSV were detected by using the developed FQRTPCR method. Compared with common PCR assay, the FQRTPCR method is more sensitive and more suitable for detection of egg drop syndrome virus in vivo and in vitro.

Analysis of Gene Expression of ALB in Duck Tissues Infected Duck Hepatitis Virus

LI Xiu;BI Yulin;XU Qi;ZHAO Wenming;ZHANG Yang;CHEN Changyi;CHEN Yang;HUANG Zhengyang;ZHEN Ting;DUAN Xiujun;CHEN Guohong

2012, 43(5): 773-778. doi:

Abstract ( 399 )

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This research tried to detect the relative expression level of mRNA and protein of ALB gene in livers, pancreases, lungs, kidneys, cerebra, cerebella, leg muscles and thymuses infected by duckling hepatitis virus (DHV1). The expression level of ALB gene and the content of ALB protein of control group, susceptible group and resistant group were detected by realtime quantitative RTPCR and ELISA techniques. Generally speaking, except in leg muscles, the expression level of ALB genes mRNA in susceptible group was highly significantly less than those in control group and resistant group (P<0.01), and the level in resistant group was significantly or highly significantly less than that in control group (P<0.05 or P<0.01). And the content of ALB protein has the same differences as ALB gene's mRNA in livers, cerebella and thymuses among those three groups, but there were no significant differences in the other tissues(P>0.05). It was showed that the results of RTPCR and ELISA techniques had the same results in the indicative tissues directly connecting with duck hepatitis such as livers, thymuses, cerebella, etc. This research gets the same results as those using suppression subtractive hybridization (SSH) and makes a further step to reveal that ALB gene is the resistant gene of duck hepatitis, whose change of the expression level can be used as a marker to distinguish susceptible and diseaseresistant ducks.

Development and Application of Microsatelliteprimered PCR for Rapidly Identifying Dermatophytes from Rabbits

LI Daijun;ZHOU Yufa;LIU Jingbo;GAO Lili;ZHAO Xiaonan;CAI Yumei;CHAI Tongjie

2012, 43(5): 779-784. doi:

Abstract ( 393 )

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The study was carried out to establish an accurate and rapid method to identify dermatophytes from rabbits by microsatelliteprimered polymerase chain reaction (PCR) and NTSYSpc2.10 software. The DNAs of Trichophyton mentagrophyton, Microsporum gypseum and Microsporum canis were amplified by the primer (GACA)4, and then their DNA fingerprint polymorphism was analyzed using NTSYSpc2.10 software. Based on the above results, the identification method was confirmed. Finally, a total of 21 clinical isolates identified by morphology were identified by the method. The results showed that: 1) The three kinds of rabbit dermatophytes have obviously different DNA fingerprints; 2) The rabbit dermatophytes of the same kind have high genetic similarity (90%100%); 3) The identified results of clinical isolates by this method were consistent with the morphological identification results. The results indicated that dermatophytes from rabbits could be rapidly and accurately identified by the microsatelliteprimered PCR in combination with NTSYSpc2.10 software.

Expression and Localization of Wnt3a in Different Colors of Alpaca Skin

SONG Yunfei#;TIAN Xue#;LU Xuxiu;GUO Qingyun;ZHANG Danli;XIE Xiaojing;YU Xiuju;HE Junping;WANG Haidong;DONG Changsheng

2012, 43(5): 785-790. doi:

Abstract ( 363 )

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This experiment was conducted to explore the expression and localization of Wnt3a in different colors of alpaca skin. The tissues were obtained from white and brown adult alpacas skins. The comparative expression quantity of gene Wnt3a in different coat colors was analyzed by real time quantitative PCR. The protein expression and localization of Wnt3a were measured by Western blotting and immunohistochemistry. The results of QRTPCR showed that the comparative expression quantity of gene Wnt3a in brown alpacas was 2.970 2 times than that in white ones. The Western blotting results showed that the protein expression of wnt3a in brown alpaca skins was significantly higher than that in white alpaca. The positive signals of Wnt3a were found in hair bulb and outer root sheath of hair follicle, and the expression of Wnt3a was significantly different between the white and brown alpaca skin based on the average optical density (P<0.05). Our findings showed that Wnt3a may involved in the regulation of hair color formation.

Morphological Mechanism of the Superior Cervical Ganglion in the Yak (Bos grunniens) Adapting to Its Habitat

DING Yanping;LI Jialong;CHAI Erqing;WANG Jianlin;SHAO Baoping

2012, 43(5): 791-797. doi:

Abstract ( 395 )

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This experiment was conducted to study the morphological adaptations of the superior cervical ganglion (SCG) of the yak to its habitat as well as investigate the adaptive mechanism of sympathetic nervous system to the QinghaiTibet plateau in plateau animals. The morphological characteristic of SCG of the yak was compared and analyzed with the cattle by morphometry method and SPSS16.0 software with HE and Toluidine Blue stained section. The results showed that: 1) The length, width and thickness of SCG of the yak were smaller than cattle significantly (P<0.01); 2) The SCG of yak were consisted with capsule, ganglion units, neuropil, vessel and connective tissue; Compared with cattle, yak has a thinner capsule and smaller proportion of connective tissue, while tissue in ganglion was more dense; 3) In both yak and cattle, the ganglion units was consisted with ganglion cell, satellite cell, fibroblasts, capillary and neuropil, while attractive characteristic was that the number of ganglion cells, satellite cells and neuropil in unit visual field of the ganglion units of the yak were similar to the cattle (P=1), but the number of the yak’s capillary were more than that of the cattle (P<0.01); 4) Similar to cattle, the head of SCG was mainly composed with ganglion units and tail was mainly nerve fibers in yak. These results indicated that as a yearround grazing and half wild animal, yak living in the QinghaiTibetan Plateau and its SCG have evolved with some morphological characteristics such as small volume, less ganglion cells and nerve fiber, and rich vessel and so on, that enabling them to adapt the ecotope well.

Genotype Analysis of Extendedspectrum βlactamases Produced by Salmonella pullorum

YUAN Li;PAN Yushan;WU Hua;LIU Jianhua;HU Gongzheng

2012, 43(5): 798-802. doi:

Abstract ( 423 )

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The resistant phenotypes and the genotypes of extendedspectrum βlactamases were detected in Salmonella pullorum by microdilution method and PCRsequencing. The results indicated that the isolates were high resistance to ampicillin, thirdgeneration cephalosporins, florfenicol, and enrofloxacin, although sensitive to cefepime, ceftiofur/sulbactam (2∶1), and cefoperazone/tazobactam (4∶1). Eight isolates harboured five kinds of ESBL genotypes (six TEM1, three CTXM65, one CTXM90, two OXA1 and one OXA10). To our knowledge, this study is the first to describe and name the CTXM90. The amino acid sequence of CTXM90 contains only one amino acid alteration, when contrasting to that of CTXM14 or CTXM65 (80Ala→Val, and 275Arg→Ser, respectively). Based on amino acid sequence similarities, CTXM90 is classified in the CTXM9 subgroup. The results illustrated that eight Salmonella pullorum showed serious multiresistance and carried complex extendedspectrum βlactamase genes.

Detection and Analysis of PlasmidMediated Quinolone Resistance Genes in Salmonella Isolates from Food Animals

LIN Juchun;QIN Chunhong;LAI Jing;SHU Gang;WU Congming

2012, 43(5): 803-809. doi:

Abstract ( 334 )

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To investigate the prevalence of plasmidmediated quinolone resistance (PMQR) in Salmonella isolates from food animals, susceptibility of 316 Salmonella isolates from food animals to 20 antimicrobial agents were determined by the broth microdilution method, the prevalence of PMQR genes in the isolates were detected by PCR. The results showed that 316 Salmonella isolates exhibited different resistance to 20 antimicrobial agents, 95.57% of isolates showed multiresistance; The positive rates of qnrA, qnrC, qnrD, qnrS, qepA, qnrB, aac(6′)Ibcr, oqxA and oqxB genes were 0%, 0%, 0%, 0%, 0%, 7.91%, 15.19%, 7.91% and 8.86%, respectively. oqxAB gene was the first time reported in Salmonella; 98.11% of PMQRpositive strains, which showed multipleresistance to 817 antimicrobials, harboured two or more resistant genes, qnrB & aac(6′)Ibcr was the most common in these strains; The results of PFGE displayed 53 PMQRpositive strains belonged to 5 PFGE patterns. Some strains ,which had different resistant phenotype or genotype, showed same PFGE patterns. In conclusion, 316 Salmonella isolates tested showed serious resistance to antimicrobial agents; qnrB, aac(6′)Ibcr and oqxAB genes were frequence in strains; Clonal strain was epidemic in some resistant isolates from different origins.

The Relationship between MMP2, MMP9 mRNA of Bovine Endometrium and Endometritis

WANG Guoqing;WANG Honghai;ZHANG Naisheng

2012, 43(5): 810-815. doi:

Abstract ( 369 )

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The study was conducted to investigate the role of MMP2 and MMP9 in cow endometritis. Healthy and suffering from acute purulent endometritis Chinese Holstein cows were selected as control and experimental group respectively with 10 in each group and all of the cows were 610 days postpartum. The concentration of PGF2a, Progesterone was detected by ELISA. CollagenⅠand Ⅳ in endometrium were checked by Immunohistochemistry. MMP2 and MMP9 expression were examined using realtime PCR in the endometrium. The results showed that the concentration of PGF2a in experimental group was lower than that in control group (P<0.01). The experimental group progesterone was significantly higher than the control group. The expression level of CollagenⅠand Ⅳ in endometristis was higher than the control group (P<0.01). The MMP2 and MMP9 mRNA expression in endometristis were lower than the control group (P<0.01). Conclusion indicated that the expression of MMP2 and MMP9 mRNA in cow endometristis reduced, CollagenⅠand Ⅳ degradation were blocked and endometrial cell was unable to secrete PGF2a, making the concentration of progesterone increase in blood serum. MMP2 and MMP9 mRNA influenced postpartum endometrial function and encouraged the development of postpartum endometritis.

Effect of Baihu Soup and Qinlian Liquid on the Classical Pathway of Complement Activation in Rabbit with Qifen Syndrome

ZHANG Shidong;WANG Dongsheng;WANG Xurong;LI Shihong;LI Jinyu;CHEN Jiongran;LI Hongsheng;YAN Zuoting

2012, 43(5): 816-820. doi:

Abstract ( 315 )

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The objective of this study was to compare the curative effect of Baihu Soup with Qinlian Liquid in molecular angle. The animal model of Qifen Syndrome was found through intravenous (i.v.) injection of LPS, and the model was treated with Baihu Soup and Qinlian Liquid respectively. Before and after treatment, the expression changes of serum CH50, C3 and CRP gene in liver tissue were determined by ELISA and realtime quantitative PCR respectively. The results demonstrated that complement system was activated through classical pathway. Baihu Soup can cured the animals through make genes of C3 and CRP high expression, and Qinlian Liquid plays a certain role only through upregulated the gene expression of CRP. The curative effect of Baihu Soup on animal Qifen Syndrome is superior to that of Qinlian Liquid.

Effect of Atipamezole on the Expression of Fos Protein Induced by Specificity Anesthetic for Miniature Pigs in the Rat Cerebral Cortex

YIN Baishuang;SHI Xingxing;LI Xiaobo;LI Zhiqiang;FAN Honggang;GAO Li;WANG Hongbin

2012, 43(5): 821-825. doi:

Abstract ( 317 )

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The research was conducted to investigate relationship atipamezole woke up rat anesthetized by XFM to cfos oncogene, by study on effect of atipamezole on the expression of Fos protein induced by XFM in the rat cerebral cortex. 72 SD rats were divided randomly into XFM group, XFM add atipamezole group, XFM add saline group. The content of cfos protein in cerebral cortex was measured by western blot. The Fos protein contents were obviously increased in the XFM anesthesia group as compared with 10 min after XFM administrated (P<0.01 or P<0.05). The Fos protein contents was never obviously changed after saline administrated as compared with control group (P>0.05). The Fos protein contents was significantly decreased after atipamzole administrated as compared with control group (P<0.05 or P<0.01). These results indicated that atipamezole palinesthesia action was concerned with cfos oncogene in cerebral cortex. Atipamzole attenuates expression of Fos protein induced by XFM in the rat cerebral cortex, it may be an important mechanism of atipamzole wake up rat anaesthetized by XFM.

Cloning, Tissue Specific Expression and Bioinformatics Analysis of Goat Lysosomal αAMA Gene

KONG Xiangya;LI Yi;CHENG Min;JING Xintang;LI Qinfan

2012, 43(5): 826-831. doi:

Abstract ( 383 )

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Goat lysosomal αAMA gene was amplified using RTPCR and the tissue specific expression profile, bioinformatics characteristic of αAMA were studied. Primers were designed based on the sequence of bovine lysosomal αAMA gene and were used in amplifying the goat αAMA, the tissue specific expression profile was analyzed by qRTPCR, and the bioinformatics analysis of αAMA was conducted. The results showed that CDS sequence of goat αAMA was 3 000 bp, encoding a deduced protein containing 999 amino acid residues in which the first 50 residues were signal peptide, and this nucleotide sequence of CDS and the deduced amino acids shared 95.93% and 94.79% homology with the αAMA mRNA of cattle. The qRTPCR revealed that the αAMA gene was expressed in various tissues at different levels. The expression level of this gene was higher in the lung, liver, and cerebellum. It was predicted that αAMA was belonged to Glycosyl hydrolases family 38, and composed of two conserved domains. There were thirtyfive phosphorylation sites, one phosphorylation site of specific protein kinase and nine Nglycosylation sites in αAMA protein. By SWISSMODEL homologous modeling, the predicted structure of goat was similar to the template and predictive. These results provide the theoretic basis for the mechanism research of αAMA and development of toxin antidote toward locoweed.

Tissue Expression Differentiation of SLADQA Gene in Yorkshire, Sutai and Meishan

ZI Chen;WU Zhengchang;LIU Lu;DU Zidong;HUANG Xiaoguo;;YANG Jiansheng;ZHU Guoqiang;;;WU Shenglong;;;BAO Wenbin;;

2012, 43(5): 832-837. doi:

Abstract ( 370 )

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The aim of this study was to analyse SLADQA gene expression in postweaning piglets of Yorkshire, Meishan and E. coli F18resistant resource population of Sutai by realtime PCR and to investigate the potential relationship between SLADQA gene and the different immune capacity of Chinese and exotic pig breeds, as well as the resistance to E. coli F18. SLADQA gene expression was detected in 11 tissues and the expression pattern was consitent in the three pig breeds，the expression level was the higher in lung, spleen and lymph node, the moderate in stomach, duodenum and jejunum. Meanwhile, the expression level of SLADQA gene was the highest in all tissues detected in Sutai piglets, followed by Yorkshire and Meishan. The expression level in lung, lymph node and thymus in Sutai was significantly higher than that in Meishan. In addition, there was significant difference in SLADQA gene’s expression between Sutai and Meishan pigs. From above analysis, it was concluded that SLADQA gene has important function of stimulating immune and antigen presentation in piglets’ response and physiological changes to the infection of E. coli F18.

Cloning and Sequence Analysis of Goats Endogenous Pulmonary Adenomatosis Retrovirus

ZHOU Yanxi;LI Jing;AO Weihua;LIU Xiaohui;YU Lixin;YAO Hongqiang;MA Xueen

2012, 43(5): 838-842. doi:

Abstract ( 346 )

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Three pairs of primers were designed according to the sequence of strain enJSRV20. Sequence of goats enJSRV was amplified by PCR technology and these PCR products were cloned into PMD18T vector and then sequenced . The complete genome of goats enJSRV was obtained successfully. Analysis results showed that the sequence of goats enJSRV was 93.4% similarity with enJSRV18 and 88.1% similarity with strain JS7, respectively. Phylogenetic analysis according to pro gene showed that goats enJSRV had higher homology with strain enJSRV5 .It will be useful for the study of constructing infectious clone and elucidation the function of endogenous pulmonary adenomatosis retrovirus.

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