Acta Veterinaria et Zootechnica Sinica

Study on the Relationship between Developmental Variants of PLIN Gene Expression and Fatness Traits in Chickens

ZHAO Xiao-ling;LIU Yi-ping;LUO Yi;ZHOU Yan;ZHU Qing

2009, 40(2): 149-154. doi:

Abstract ( 825 )

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In order to discover the function and mechanism that Perilipin (PLIN) gene acted on fatness traits of chicken，we adopted the real-time PCR method to test the relative expression quantity of PLIN gene in breast muscle，liver， abdominal fat and subcutaneous fat of Sichuan Mountainous Blackbone chicken at 7 growth points， including 1，14，28，42，56，70 and 84 day.We also tested each bird′s comb weight， living weight， subcutaneous fat thickness and intramuscular fat content in breast muscle. The results showed that the order of the relative expression quantity of PLIN gene in four tissues from high to low was abdominal fat， subcutaneous fat， breast muscle， and liver for all the growth points， except for 28 and 70 day. And the relative expression quantity of PLIN gene in abdominal fat and subcutaneous fat were increasing along with the growth of bird. Furthermore， the developmental variations of relative expression quantity of PLIN gene in abdominal fat were positively related with those of subcutaneous fat thickness（P≤0.01）. And the developmental variations of relative expression quantity of PLIN gene in breast muscle were positively related with those of intramuscular fat content （P≤0.01）. We inferred PLIN gene was a key regulator of the fatness traits development in chicken.

Analysis of the Polymorphisms of AIRC Gene and Its Genetic Effect on IMP Content in Chicken

SHU Jing-ting;BAO Wen-bin;WU Xin-sheng;ZHANG Xue-yu;ZHANG Hong-xia;CHEN Guo-hong

2009, 40(2): 155-160. doi:

Abstract ( 1270 )

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Single nucleotide polymorphisms (SNPs) of AIRC gene were investigated in various breeds（including Recessive White chickens， Silkies chickens， Baier chickens，Tibetan chickens and Xiaoshan chickens）by PCR-SSCP and DNA sequencing methods. Two SNPs were detected in exon 3 and 8. The least square analysis showed that birds with CC genotype had significant higher IMP content than those with TC and TT genotypes， birds with TC genotype also had a little higher IMP content than those with TT genotype， but the difference was not significant in exon 3; In exon 8， CG was the excellent genotype， individuals with CG genotype had significant higher IMP content than those with GG and CC genotypes， birds with GG genotype also had significant higher IMP content than those with CC genotype. The combined genotypes of exon 3 and 8 also had significant genetic effect on IMP content. The IMP content of the individuals with CCCG combined genotype was 2.25 mg·g-1 higher than the individuals with TTCC combined genotype. The genetic effect of the combined genotype was larger than that of excellent single genotype. Therefore， it was concluded that the combined genotypes could be used as the molecular genetic marker to select the chicken for meat quality traits.

Tissue Distribution, SNP Detection and Its Association Analysis with Carcass and Meat Quality Traits of Chicken MyoT Gene

ZHU Zhi-ming;CHEN Guo-hong;QIANGBA Yang-zong;LI Chang-chun;ZHU Meng-jin;FAN Bin;LIU Bang

2009, 40(2): 161-165. doi:

Abstract ( 1497 )

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The semi-quantitative RT-PCR was performed to detect the expression pattern of chicken MyoT gene in 7 different tissues (heart, liver, spleen, lung, kidney, brain and muscle). The results showed that MyoT gene was expressed only in heart and muscle. Partial DNA sequence of chicken MyoT gene was cloned and a A/G transition was found in exon 10. Using PCR-Hin6Ⅰ-RFLP method, the A/G SNP locus was analyzed in 12 native chicken breeds (White Ear, Xianju, Dagu, Luyuan, Gushi, Game chickens, Silky Sooty, Beijing Fatty, Chahua, Xiaoshan, Langshan and Tibetan chicken) and 1 foreign breeds (Recessive White chicken). The association between different genotypes and carcass and meat quality traits was analyzed in Tibetan chicken, Recessive white, hybrid chicken (Tibetan♂×Recessive♀). The result of association analysis showed there were correlation between this SNP site and leg muscle weight (P＜0.05), and significant correlation between this SNP site and leg muscle rate and meat colour (leg muscle) (P＜0.01).

Study on Association of Restriction Fragment Length Polymorphisms of Fatty Acid Synthase Gene with Carcass， Liver Performance and Fat Deposit Traits in Landes Goose

KUANG Zhi-xiang;HE Da-qian;LIU Yi-ping;ZHU Qing

2009, 40(2): 166-172. doi:

Abstract ( 1337 )

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The aim of this study was to investigate polymorphism of fatty acid synthase (FAS) gene promoter region in Landes goose. Polymorphisms were detected by restriction fragment length polymorphisms(RFLP) and enzyme AfaⅠwas used. Three AfaⅠdigestion sites (locus A， B and E) were detected in Landes goose. The χ²-test was performed to examine the HardyWeinberg equilibrium (HWE). Three sites significantly deviated from HWE. The least square analysis showed that living weight， weight gain at feeding increase stage， carcass weight， halfeviscerate weight， eviscerate weight， breast muscle weight and leg muscle weight of individuals with genotype FAS^EE were higher than that of individuals with genotype FAS^ee 12.78%-19.67% （P＜0.05） at loci E and B. Locus A had no significant effects on carcass traits except for breast muscle weight（P＞0.05）. Fatty liver performance of individuals with FASAa and FAS^Bb were better than that of individuals with homozygote at loci A and B， but the liver weight of individuals with genotype FAS^EEwas higher than that of individuals with heterogenesis 33.09％（P＜0.05）at locus E. Three loci had no significant effects on fat deposit traits.

Analysis of Genetic Structure in Wild Boar and Their Crossbred Populations

LIU Li-hua;MIN Ling-jiang;SHEN Wei;PAN Qing-jie

2009, 40(2): 173-179. doi:

Abstract ( 885 )

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The genetic polymorphism and differentiation were analyzed in Wild boar， Duroc，Landrace and crossbred populations of Wild boar×Duroc and Wild boar×Landrace with five structural genes by PCR-RFLP and PCR-SSCP.The results showed as follows: (1) In the above detected populations，the mean heterozygosity and PIC indicated that Wild boar presented low polymorphism while other populations had mediate polymorphism.(2) The results indicated that 17.9% and 82.1% of total variation were respectively from between and within three populations including Duroc， Wild boar and Wild boar×Duroc， while 27.1% and 72.9% of total variation were respectively from between and within three populations including Landrace， Wild boar and Wild boar×Landrace， so the genetic differentiation in Wild boar，Duroc，Landrace and their crossbred populations were at the mediate level(Fst＞0.15). (3) Distance of Reynolds′ indicated that Duroc and Wild boar with larger genetic distance will bring greater heterosis than Landrace and Wild boar.

Effects of Data Preprocessing and Measuring Metrics for Different Gene Expression Data

LIU Tian-fei;TANG Guo-qing;LI Xue-wei

2009, 40(2): 180-184. doi:

Abstract ( 768 )

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The effects of different measuring metrics and data preprocessing for different gene expression data on K-means clustering were studied. The results illustrated that different data preprocessing ways made significant differences under different measuring metrics. The best data preprocessing in K-means clustering was to select log transformations for the timecourse gene expression dataset， and measuring metrics is to select covariance metrics. However， the best data preprocessing is log transformations for other datasets， three measuring metrics (Euclidean distance， squared Euclidean distance and Manhattan distance) led to better results.

Multiplex PCR Detection of Exogenous Gene in Transgenic Mice

WANG Chen-fang;LI Xin-yun;WANG Zhi-wei;AO Hong;CUI Wen-tao;LI Kui

2009, 40(2): 185-190. doi:

Abstract ( 1204 )

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Focused on detection method for transgenic animals and its standardizations abroad in this article.It is essential to administrative regulations of transgenic animal and label of transgenic animal food.Extracting genomic DNA from mice tails by alkaline lysis method.The positive mice were screened by detecting of several transgenic elements such as cytomegalovirus (CMV) promoter， hGH polyA terminator， transmembrane protein 66 (Tmem66)target gene with multiplex PCR method， as detecting mice endogenous GAPDH gene. The process of the multiplex PCR reaction was optimized，such as the optimal annealing temperature and the concentration of the PCR buffer， simultaneously studying the effect of reaction rate on multiplex PCR.The results showed that the multiplex PCR method was simple and accurate to detect transgenic elements of transgenic mice， which offered us useful experience for developing the typical testing standard of transgenic mammals and its drived food.

Experimental Research on Efficacies of GnRH-A Immunocastration in Rabbits

WEI Suo-cheng;GONG Zhuan-di

2009, 40(2): 191-196. doi:

Abstract ( 1376 )

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To investigate the efficacies and mechanisms of immunocastration by using gonadotropin releasing hormone analogues (GnRH-A) in animals. The GnRH-A antigen emulsion was made by adding incomplete Freund′s adjuvant after GnRH-A was combined with bovine serum albumin(BSA). 30 rabbits were injected with GnRH-A antigen, the testis length and weight as well as body weight of the immunized rabbits were measured，GnRH antibody titer and testosterone concentration were detected through ELISA. There was a significant difference of the testis lengths between two EGs groups(P<0.01)，antibody levels of EG-Ⅱ group was higher obviously than that of EG-Ⅰ; the serum testosterone concentration between EG-Ⅰ and EG-Ⅱ groups were remarkably different from 7 to 16 weeks(P<0.05 )，the body weight and average daily gain of EG-Ⅱ group were the biggest and significantly higher than that of EG-Ⅰ group as well CG(P<0.05). The testis development，serum GnRH antibody titer and testosterone concentration of experiment rabbits changed obviously after immunization of GnRH-A and adding another injection would strengthen castration effects.

Effects of Different Dietary Crude Protein Level on the Expression of IGF-Ⅰ mRNA in Adipose and Muscle Tissue of Sheep

LIU Jing-yun;ZHANG Ying-jie;LIU Yue-qin

2009, 40(2): 197-202. doi:

Abstract ( 739 )

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Twenty four crossbred ewes (BlackSuffolk♂× Small Tail Han sheep♀) were randomly allocated into three treatments with different dietary CP levels (high protein， medium protein and low protein) respectively. The expression level of IGF-Ⅰ mRNA in adipose tissue and muscle of sheep were quantified by RT-PCR. The results showed that expression of IGF-Ⅰ mRNA increased with the increase of dietary CP level in abdominal subcutaneous adipose tissue， mesentery adipose tissue and semitendinosus muscle of sheep. The effects of dietary CP level on IGF-Ⅰgene expression showed tissue specificity. For abdominal and mesentery adipose tissues， IGF-Ⅰ mRNA expression increased with the increase of dietary CP level (P<0.05)， while the expression of IGF-ⅠmRNA in semitendinous muscle was significantly elevated only when the high protein diet was fed (P<0.01) and the difference of IGFⅠmRNA level between the medium and high protein treatments and between the medium and low protein treatments was insignificant (P>0.05).

Effects of High Ambient Temperature on Meat Quality and Flavor in Commercial and Local Broilers

LU Qing-ping;WEN Jie;ZHANG Hong-fu;DONG Ya-wei;WANG Qi-jun

2009, 40(2): 203-207. doi:

Abstract ( 1455 )

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108 healthy male chickens (5 wk old) with similar body weight，from Arbor Acres (AA) and Beijing-You (BJY) respectively were used to study the effects of chronic heat stress on meat quality and contents of the flavor substances in breast muscle. Chickens from each stock were randomly alloted into 3 treatments: constant high ambient temperature at 34 ℃ with ad libitum feeding (34AL)， constant optimal ambient temperature at 21 ℃ with ad libitum feeding (21AL)， and constant optimal ambient temperature 21 ℃ but pair-fed to the amount consumed by the 34AL group (21PF). All birds were reared in 3 environmental chambers until the end of the trial at 8 wk of age. The results showed that exposed to the heat of 34 ℃ for three weeks， AA broilers exhibited higher L* values and drip loss in breast muscle(P<0.05)， moreover， they showed decreased content of insine monphosphate (IMP) (P<0.01). For BJY chickens， heat exposure had no significant influence on meat quality traits (except for pHu) and contents of IMF and IMP in breast muscle (P>0.05). The data indicated that chronic heat exposure (34 ℃) had a detrimental effect on meat quality traits of breast muscle in AA broilers， while BeijingYou chickens had higher adaptability to high temperature， their muscle fibers were insensitive to heat exposure (34 ℃).

The Effects of High-fat Diet on the Digestion， the Absorption and the Level of Free Radical Produced in the Mice Digestive System

WANG Jiao;YUE Peng;WANG Yan-yan;SHI Yong-hui;WANG Zhou-ping;LE Guo-wei;

2009, 40(2): 208-212. doi:

Abstract ( 1574 )

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Forty male mice(6-week-old) were randomly assigned into four groups according to the body weight. The control group， the high-fat diet group (crude fat contain 18%)， the LA 1 group (highfat diet with 0.05% LA) and the LA 2 group (high-fat diet with 0.1% LA). After 5 weeks feed， the metabolism experiment was carried out. 6 weeks later， the level of the free radical in organizations of the digestive system and the activity of protease， lipase and amylase in the digestive tract were detected. The results showed that feeding the mice with high-fat diet in longterm could improve the level of the free radical in the mice digestive system (P<0.05) and the activity of the lipase in the digestive tract significantly (P<0.05)， decrease the activity of the protease and amylase (P<0.05)， decline the digestibility of the fat， protein (P<0.05) and the sedimentation rate of the protein (P<0.05). Adding 0.05% LA into the high-fat diet could incline the level of the free radical remarkably， enhance the activity of these enzymes and digestibility of the fat，protein and the sedimentation rate of the protein(P<0.05). Adding 0.1% LA into the high-fat diet can recover the level of free radical up to normal level.It was concluded that the high-fat diet could significantly increase the production of the free radical in the mice digestive system，decrease the activity of the digestive enzymes and the absorption of the nutrition. While adding the optimum LA into the high-fat diet could get rid of these disadvantage and relieve the oxidative stress.

Construction and Identification of the Full-length Genomic cDNA Clone of FMDV China99/S Isolated from Swine

CHANG Yan-yan;ZHENG Hai-xue;JIN Ye;WANG Guang-xiang;SHANG You-jun;LIU Xiang-tao

2009, 40(2): 213-220. doi:

Abstract ( 1337 )

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Total RNA was extracted from tissue samples of FMDV China99/S strain and four overlapping fragments (A， B， C and E) were amplified. The pP_ⅠcDNA3.1（+）vector with RNA polymerase Ⅰand A， B， C， E fragments were digested respectively by restriction endonucleases(A(BsmBⅠ/XmaⅠ)，B(XmaⅠ/ BamHⅠ)， C(HpaⅠ/KpnⅠ)，E(BamHⅠ/NotⅠ))， and then ligated to produce recombinant plasmid of pP_ⅠFMDVcDNA3.1（+）. The analysis of nucleotide sequences showed that the genome of swine FMDV China99/S strain is 8 116 nucleotides in length(including Poly(C) and Poly(A) tracts)， and the open reading frame (ORF， 6 318 nucleotides which encode 2 106 amino acids) is flanked by a 5′non-coding region (5′NCR) of 1 064 nucleotides (including 41 C residues Poly(C) tract) and a 3′NCR of 131 nucleotides (including the Poly(A) tail at least 38 A residues). Recombinant plasmid of pP_ⅠFMDVcDNA3.1（+） was transferred into BHK-21 cells with liposome and typical FMDV pathological changes were observed in subculture. The electron microscope,CPE,animal test,RT-PCR and sequencing analysis verified that FMDV China99/S was rescued.

Cloning and Sequence Analysis of the HA，NP，NA，M and NS Genes of H1N2 Swine Influenza Viruses

MENG Xue-qiong;CHEN Yi-xiang;LIU Qi;ZHENG Min;SHI Kai-chuang;HU Jie

2009, 40(2): 221-227. doi:

Abstract ( 1388 )

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In this study，the HA， NP， NA， M and NS genes of three isolates of H1N2 swine influenza viruses (SIV) were cloned and sequenced. Sequence comparisons revealed that three isolates share a higher degree of similarity. Genetic analysis demonstrated that three isolates were closely related to the United States triple reassortant H1N2 SIV isolate. In the HA， NP， M and NS genes phylogenetic trees， three isolates clustered with classical H1N1 SIV. While in the NA gene tree， isolates clustered with human influenza viruses. Comparisons of deduced amino acid sequences of HA and NA of the 3 isolates with the representative viruses A/swine/Maryland/23239/1991 (H1N1) and A/Buenos Aires/4459/96 (H3N2) revealed that they shared 954%961% (HA) and 966%972% (NA) amino acid similarities， respectively. Amino acid substitutions were detected at previously defined glycosylation sites， antigenic sites and receptor binding sites of the HA or NA proteins. The effects of isolates owing to these amino acid substitutions on biological characterization are unknown.

Effects of Porcine Reproductive and Respiratory Syndrome Virus and Porcine Circovirus Type 2 Co-infection on Virological Kinetics and Immunological Responses in Piglets

SHI Kai-chuang;YANG Han-chun;GUO Xin;HE Wan-lun;GE Xin-na;CHEN Yan-hong;ZHA Zhen-lin

2009, 40(2): 228-234. doi:

Abstract ( 1526 )

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To explore the interaction between porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) in pathogenesis， the virological kinetics and immunological responses in piglets to PRRSV and PCV2 co-infection were analyzed. Twenty healthy， six-week-old piglets were randomly divided into control group， PRRSV or PCV2 infected group and PRRSV/PCV2 co-infected group with 5 piglets each. After infection， serum and different tissue samples， including heart， liver， lung， kidney， pancreas， spleen， thymus， tonsil， inguinal lymph node and mesenteric lymph node， were used for PRRSV and PCV2 quantitation by real-time PCR， and serum was used for PRRSV- and PCV2-specific antibody detection by indirect ELISA and indirect IFA， respectively. The proliferation activities of peripheral blood lymphocytes (PBLC) were detected by 3-[4，5dimethylthiazol]-2，5-diphenyltetrazolium bromide (MTT). The results showed that PRRSV- and PCV2-loads in both serum and different tissue samples from co-infected group were significantly higher than those of PRRSV or PCV2 singly infected group. Antibodies to PRRSV and PCV2 developed later and reached significantly lower levels in serum from PRRSV/PCV2 group than from PRRSV or PCV2 group. The proliferation activities of PBLC in three virusinfected groups were suppressed and the degree of severity was shown as follows: PRRSV/PCV2 group > PRRSV group > PCV2 group. These results indicate that PRRSV and PCV2 co-infection can promote the replication of both PRRSV and PCV2 in vivo and synergistically suppress the immune responses of the co-infected piglets.

Sequence Analysis and Phylogenetic Relationships of 11 Mite Isolates Based on ITS-2 Gene

GU Xiao-bin;ZHANG Xiao-qian;YANG Guang-you;JIA Xiao-yong;WANG Shuai

2009, 40(2): 235-242. doi:

Abstract ( 1460 )

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In order to clarify the taxonomic status of several species of mites which are common causative agents of mange in many animal populations， the second internal transcribed spacer (ITS-2) of the rRNA gene was sequenced and compared with the homologous sequences of 6 Sarcoptes isolates， 5 Chorioptes isolates， and 21 sequences of mites retrieved from GenBank. The phylogenetic relationships were reconstructed basing on 32 isolates using UPGMA method. The length of ITS-2 was 361 bp in 6 Sarcoptes isolates. In these six nucleotide sequences， a total of 335 nucleotide positions were conserved， 15 were variable sites， and 9 were parsimony informative sites. Homology analyses indicated that the identity levels of nucleotide of ITS-2 among the 6 isolates ranged from 96.9% to 99.7%. The length of ITS-2 in 5 Choriptes isolates were different (225.232 bp). The conserved sites， variable sites， parsimony informative sites were 184， 44 and 9， respectively. The sequence identity of Chorioptes mites from four kinds of cattle or cow ranged from 92.4% to 97.3%， the one between Chorioptes mite from Giant panda and other 4 isolates ranged from 78.7% to 82.6%. A phylogenetic tree constructed using the UPGMA method indicates that the mites examined in this study are clustered in the second major clade corresponding to their second families. The first clade includes the genus of Sarcoptes and Notoedres， the second clade is comprised of the genus of Chorioptes and Psoroptes. According to the homology analysis and phylogenetic analysis， we support the view that the genus Sarcoptes consists of only one species， and the taxonomic status of Chorioptes isolates from Giant Panda and the Psoroptes isolates from buffalo are still to be verificated furtherly.

Pharmacokinetics/Pharmacodynamics Integration of Florfenicol against E. coli in Pigs ex vivo

YANG Yu-hui;YANG Dong;DING Huan-zhong;ZENG Zhen-ling

2009, 40(2): 243-247. doi:

Abstract ( 1386 )

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For the rational usage of florfenicol to treat the colibacillosis of pigs，the integration of pharmacokinetics in vivo and pharmacodynamics in vitro was used to study the antibacterial activity of florfenicol against E.coli in pig serum ex vivo. The accurate MIC of florfenicol against E. coli in MH broth and serum was 3.25 and 8.75 μg·mL^-1， respectively. Florfenicol was absorbed slowly and anomaly in pigs after intramuscular administration（20 mg·kg^-1）. The Cmax was (5.28±1.48) μg·mL^-1 at (3.60±1.52) h. The elimination of florfenicol was slow in pigs. Mean residence time(MRT) was (26.61±9.81) h. Elimination halflife(t1/2β) was (17.49±8.04) h. EC50 was (7.76±4.53) h. AUC0→24/MIC was (7.69±1.48) h and Cmax/MIC was (0.60±0.17). Because of the poor activity of florfenicol against E.coli and the lower plasma Cmax of florfenicol in pigs， it may fail to get desirable treatment result when curing the infection of E. coli in pigs by the routine regimen.

Study on the Expression and Bioactivity of Porcine Interferon Alpha/Porcine Interleukin-2 Fusion Protein

YAN Ruo-qian;WU Zhi-ming;ZHANG Zhi-ling;SHENG Min;LIU Guang-hui;ZHAO Ming-jun

2009, 40(2): 248-255. doi:

Abstract ( 1387 )

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To exploit the high efficient porcine gene engineering antiviral agent to prevent and control the porcine viral disease， the recombination chimeric gene of PoIFN-α-linker-PoIL-2 was constructed by linking porcine interferon alpha (PoIFN-α) mature peptide gene and porcine interleukin-2 (PoIL-2) mature peptide gene via a 15-amino acid glycine-rich linker [linker，（G₄S）₃] by SOEPCR (splicing by overlap extension-PCR) method. Then the recombinant gene was cloned into pGEM-T Easy vector and subsequently sub-cloned into prokaryotic expression vector pQE30. The recombinant PoIFN-α-linker-PoIL-2 (rPoIFN-α-linker-PoIL-2) protein was expressed in E.coli JM109 and purified under the process of denaturing by 8 mol·L^-1 urea， refolding by a self-innovative renaturation buffer and dialyzing by PBS buffer etc. The antiviral bioactivity of rPoIFN-α-linker-PoIL-2 protein was tested by inhibiting the 50 percent appearance of cytopathic effect (CPE) of vesicular stomatitis virus (VSV) on porcine kidney 15 (PK15) cell lines and highpathogenic porcine reproductive and respiratory syndrome virus (PRRSV) on Marc-145 cell lines. The PoIL-2 bioactivities of rPoIFN-α-linker-PoIL-2 protein were estimated by the methods of IL-2 MTT assay and porcine IL-2 ELISA assay for detection PoIL-2 protein，respectively. The results showed that the correct PoIFN-α/PoIL-2 fusion protein had been obtained. The antiviral activity of rPoIFN-α-linker-PoIL-2 protein to inhibit the reproduce of VSV on PK15 cell line was 1.891×10⁴ IU·mL^-1， and the lowest effective dose of rPoIFN-α-linker-PoIL-2 protein to inhibit the proliferation of high-pathogenic PRRSV on marc-145 cells was 905 U·mL^-1， which were similar to the antiviral activity of recombinant PoIFN-α protein (rPoIFN-α) control. The rPoIFN-α-linker-PoIL-2 protein displayed the ability to stimulate the proliferation of IL-2-dependent CTLL-2 cells and the specific immune response for monoclonal antibody (MAb) of PoIL-2 by ELISA assay respectively，which were similar to the bioactivity of recombinant PoIL-2 protein (rPoIL-2) control. Our study indicated that the rPoIFN-α-linker-PoIL-2 protein had the duplex bioactivity of PoIFN-α protein and PoIL-2 protein on cells，which laid a foundation for further wide using the rPoIFN-α-linker-PoIL-2 protein as the main ingredient of porcine gene engineering antiviral agent to prevent and control the high-pathogenic porcine reproductive and respiratory syndrome and other porcine viral diseases.

Effects of High Dietary Selenium on the Proliferation Function and Subset of Peripheral Blood T Lymphocyte in Chickens

PENG Xi;CUI Yun;YANG Fan;CUI Wei;CUI Heng-min

2009, 40(2): 256-260. doi:

Abstract ( 767 )

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By the methods of FCM and MTT, the subsets and lymphocyte proliferation function of peripheral blood T lymphocyte were studied in AA poultries by feeding with sufficient (0.2 mg·kg^-1, control group), or excess selenium (1, 5, 10, and 15 mg·kg^-1), as sodium selenite in the diets to five groups of 60 chickens each for 6 weeks. The results showed that the percentage of CD3⁺ T cells, CD3⁺CD4⁺ T cells and CD3⁺CD8⁺ T cells were decreased variably in chickens when dietary selenium exceeded 5 mg·kg^-1. The mitogenic response of T lymphocytes to ConA and the content of serum IL-2 were decreased in high Se group Ⅲ and high Se group Ⅳ (P<0.05 or P<0.01) when compared with control group. The results suggested that the dietary selenium in excess of 5 mg·kg^-1 impaired the cellular immune function in chickens.

Effects of Aluminium Intoxication on Growth and Morphological Structure of Spleen and Bursa of Fabricius in Chickens

LIU Fu-tang;HUANG Jing-feng;GU Qing-yun;LI Yan-fei

2009, 40(2): 261-265. doi:

Abstract ( 1257 )

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Model of chickens with deuto-chronicity different level of AI（Aluminium intoxication） toxic was established by continuous peritoneal injection of fixed volume and different concentration gradient aluminium trichloride. The growth index of spleen and bursa of Fabricius were detected，and the structure of them were observed by using light and electron microscope.The results showed that:(1)The growth index of spleen and bursa of Fabricius were significantly lower in chickens with AI toxic than that in the control ones and there were dose-effect relationship with aluminium trichloride.(2)Histopathologically，middle and low dose group had no apparent pathological change，in high dose group，spleen presented changes of apoptosis morphology and germinal center of folliculus lymphaticus in bursa of Fabricius hyperplasyed.(3)Ultrastructurally，low dose group had no apparently pathological change，in high dose group，karyolemma broke，mitochondria swelled，cristae broke or disappeared，rough endoplasmic reticulums dilated，cisternas widen，confluenced to vacuole and presented typical mark shape of apoptosis in lymphocyte of spleen and karyolemma broke，caryolysised，myelin bodys appeared，cell organs of intracytoplasm decreased obviously in lymphocyte of bursa of Fabricius.The results indicated that AI toxic greatly inhibited the development of spleen and bursa of Fabricius，and caused obvious pathological injury in these tissues.

Effects of Psoralea corylifolia L. Extracts and Psoralen on Osteoblasts Cultured in vitro

AN Ya-lan;WANG Jian-fang;XU Shui-ming;HAN Bo

2009, 40(2): 266-271. doi:

Abstract ( 1209 )

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In order to evaluate the effect of Psoralea corylifolia L. extracts and Psoralen with different concentrations on osteoblasts cultured in vitro. The proliferation and the activity of alkaline phosphatase（ALP）were measured by MTT(tetrazolium salt test) and PNPP(P-nitrophenyl phosphate disodium)， respectively，and the cell-cycle analysis was examined with flow cytometer by PI(propidium iodide) staining. The results indicated that 10 and 10^-3 mg·mL^-1 of Psoralea corylifolia L. extracts could remarkablely promote osteoblasts proliferation. Psoralen at lower concentrations（10^-7，10^-8 mol·L^-1） could promote proliferation of osteoblasts， while it could inhibit proliferation at higher concentrations（10^-4 ，10^-5 mol·L^-1）. ALP activity was significantly increased by different concentrations of Psoralen. The percentage of G1 phase cells was decreased，while the S and the G2 phase were increased. It is suggested that Psoralea corylifolia L. extracts and Psoralen，which promote the proliferation of osteoblasts with a phytoestrogenic effect， can also induce the transformation from the G1 phase to either the S phase or the G2 phase. Psoralen may play an important role in differentiation of osteoblasts.

Analysis of Genetic Diversity of Mingguang Small-ear Pig

HUO Jin-long;MIAO Yong-wang;HUO Hai-long;LI Da-lin;SHEN Xue-ying;QU Zai-jiu;YIN Yu-an

2009, 40(2): 272-279. doi:

Abstract ( 753 )

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In order to reveal the genetic variation in Mingguang small-ear pig population and provide the genetic background for its preservation and utilization， 65 individuals sampled from the breed were assayed by using 76 microsatellite loci located on swine′s 19 pairs of chromosomes. A total of 343 alleles were detected， the number of alleles varied from 2 to 9 and the number of effective alleles from 1.223 9 to 4.807 9， giving mean allele number at each locus was 4.513 2±1.205 5 and effective allele number was 3.216 9±0.773 6. The mean observed heterozygosity， expected heterozygosity and polymorphism information content(PIC) in the population were 0.944 2±0.159 5， 0.668 5±0.094 5 and 0.610 3±0.108 3， respectively. The results indicated that the genetic variation was rich in this population.

Preliminary Confirmation of Culex tritaeniorhynchus Carry with Porcine Reproductive and Respiratory Syndrome Virus

SHI Kai-zhi;MAO Jun-ting;WANG Kai-gong;ZHOU Bi-jun;WEN Ming;LI Yong-ming

2009, 40(2): 280-284. doi:

Abstract ( 1345 )

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The NSP2 gene fragments of porcine reproductive and respiratory syndrome virus (PRRSV) were amplified by RTPCR from the extracted RNA samples of Culex tritaeniorhynchus collected from 3 infected pig farms， and then were cloned into pMD-18T vector for sequence analysis. And the homogenate supernatant of Culex tritaeniorhynchus samples were inoculated to the Marc-145 cells for isolation and identification of PRRSV. The results showed that the specific DNA fragments of about 1 060 bp were amplified from the RNA samples from Culex tritaeniorhynchus， and the sequences of these fragments shared high homology with the NSP2 gene of PRRSV isolates from infected pigs in the same farm. And there are 90 nucleotides deletion in NSP2 gene compared with CH-1a strain of PRRSV. The obvious cytopathic effect were appeared in Marc-145 cells inoculated with the samples of Culex tritaeniorhynchus， and the specific fluorescence were observed in the infected cells by indirect immunofluorescent assay， and the PRRSV-N gene fragments were amplified by RT-PCR from infected cells. These results indicated there are PRRSV in Culex tritaeniorhynchus， and it could be an important way of prevalence and transmission of PRRS.

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