Acta Veterinaria et Zootechnica Sinica

Changes of Superoxide Dismutase Activity and Malondialdehyde Level in PostmortemMuscle and Their Association with Meat Quality in Pigs

LI Hua;ZENG Yongqing;WEI Shudong;CHEN Qimei;SONG Yiping;QIAN Yuan;DONG Bin;CUI Zhifeng

2010, 41(3): 257-261. doi:

Abstract ( 786 )

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Laiwu (n=24), Lulai Black (n=24) and Yorkshire (n=12) castrated boars were used to analyze the changes of superoxide dismutase (SOD) activity and malondialdehyde (MDA) level in postmortem muscle and their association with meat quality. The results showed as follows: ① As storage prolonging, SOD activity in postmortem muscle tended to decrease, but MDA level tended to increase. The breeds had significant effects on SOD and MDA (P<0.01), except for MDA5 d (P>0.05). For different pig breeds, the order of SOD activity was Laiwu > Lulai Black> Yorkshire, but MDA level was Laiwu <Lulai Black< Yorkshire. ② For meat quality, different breeds had significant effects on meat color, drip loss, water holding percentage, cooking loss, intramuscular fat (IMF), polyunsaturated fatty acids (PUFA) (P<0.01). Compared with Yorkshire, Laiwu and Lulai Black pigs were bright red in meat color, higher in water holding capacity (WBC) and content of IMF, but lower in content of PUFA. ③ SOD activity and MDA level in postmortem muscle, to some extent, were highly correlated to WBC, meat color and shear force. It proved that the muscle with higher SOD activity and the lower MDA level had the better meat color, WBC and tenderness. It implied that SOD decided the characters of meat quality by quenching oxygen free radicals and inhibiting lipid oxidation in muscle.

A Typing System Based on Ligase Detection Reaction for Chicken Meat Flavor Related Candidate Genes Polymorphism

LI Huifang;SHU Jingting;ZHANG Ying ;KUANG Zhixiang;ZHU Wenqi;HAN Wei;CHEN Kuanwei

2010, 41(3): 262-267. doi:

Abstract ( 670 )

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To establish a parallel typing system based on ligase detection reaction (LDR) detecting the polymorphisms of AFABP, ExFABP and ADSL genes, which were all candidate genes for chicken meat flavor, and to analyze the distribution differences of different genotypes in Qingyuan partridge chickens and Recessive White chickens, 165 hen samples and 185 hen samples were collected from Recessive White chicken and Qingyuan partridge chicken, respectively. The genotypes of AFABP 51C/T, ExFABP 1011T/C and ADSL 3484C/T were identified by using PCRLDR. The results by this method were coincidence with direct sequencing. Three SNPs all exhibited medium polymorphism. The allele distribution in different breeds was different, and the difference in AFABP 51C/T locus was significant. The results indicated that the parallel typing system based on ligase detection reaction is a accurate and sensitive method to detect SNPs, and will lay the research foundation for the polymorphisms of candidate genes relating to meat flavor.

Study on the Relationship between Polymorphism of PLIN Gene and Carcass and Meat Quality Traits in Qinchuan Cattle

FAN Yueyuan;ZAN Linsen;WANG Hongbao;YANG Yanjie

2010, 41(3): 268-273. doi:

Abstract ( 807 )

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To reveal the polymorphism of exon3 and exon4 of PLIN gene in Qinchuan beef cattle and evaluate its relationship with production traits. 405 Qinchuan cattle with 1824 months age under identical feeding condition were selected. PCRSSCP and sequencing method were applied in this study to detect the SNPs of PLIN exon3 and exon4, and analyze the correlation of SNPs with carcass and meat quality traits. T→C mutation at 156 base site in exon3 and C→T mutation at 14 base site in exon4 of this gene were detected. Correlation analysis showed that the index about slaughter weight, carcass weight, backfat thickness and loin muscle area of individual with AB genotype were significant higher than those with AA and BB genotypes(P<0.05), and compared to individual with AA genotype was very significantly different(P<0.01). This analysis also showed that the carcass length, carcass chest depth and backfat thickness of individual with DE genotype were significant higher than those with EE and DD genotypes(P<0.05), and compared to individual with EE and DE genotypes, the tenderness of individual with DD genotype was very significantly different(P<0.01). The results indicated that the SNPs of PLIN gene had effect on the capacity of fat accumulate of Qinchuan cattle,it could be a candidate gene for beef modify and directed to the beef cattle breeding.

Cloning and Analyzing the Promoter of Mouse TLE4 Gene

ZHAO Lili;MU Yulian;CUI Wentao;YANG Shulin;TANG Zhonglin;LI Yong;ZHAO Weimin;LIU Di;LI Kui

2010, 41(3): 274-278. doi:

Abstract ( 737 )

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The preliminary objective of the study was to investigate the transcriptional regulatory mechanisms of mouse TLE4.5′upstream promoter sequences (spanned from -2 521 bp to +327 bp) of mouse TLE4 gene were amplified by PCR. Seven promoter fragments with different length were obtained by walking deletion and then cloned into luciferase report gene expression vectors. The vector expression activities were determined by transfection of the mouse F9 teratoma cells and the mouse embryonic stem cells with the constructed dualluciferase vectors.The experiment results indicated that negative regulation elements were localized within the promotor region from -2 521 bp to -2 137 bp, and further deletion analysis indicated that the region from -2 027 bp to -1 927 bp in TLE4 gene promotor showed higher activity than other regions. In addition, a functional HSF regulation element was identified in this region. The results inferred that the HSF played an important role in the regulation of TLE4 gene expression and its functional performance.

Transient Transfection Factors for Expression of Recombinant Somatostatin Plasmid pEGS/2SS in Cultured HeLa Cells

HE Xiaohong;CAO Shaoxian;MAO Dagan;YANG Liguo

2010, 41(3): 279-285. doi:

Abstract ( 700 )

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We aimed to investigate the factors infecting transfection efficiency of somatostatin recombinant plasmid pEGS/2SS introducting into HeLa cells, make preparations for advanced study on somatostatin gene vaccine mechanism and effect of action. Four parameters including transfection reagent (lipofectamineTM) concentration, DNA concentration, cells cultured time after transfection and DNA extracting techniques were analyzed and optimized, and transfection efficiency was evaluated by the percentage of transfected cells with green fluorescence under fluorescence microscope. Based on the optimized lipofectamineTM and DNA concentration, 34.02% transfected HeLa cells could express green fluorescence 72 h after transfection when the concentration of lipofectamineTM and DNA was individually 6 μg and 1 μg; Based on the optimized cells cultured time after transfection and DNA extracting techniques, 17.88% transfected HeLa cells could express green fluorescence protein with Plasmid Maxi Kit extracting DNA 48 h after trasfection. Optimal transfection conditions were determined with four parameters, 17.88% HeLa cells could express green fluorescence protein with Plasmid Maxi Kit extracting DNA, 1 μg DNA and 6 μg lipofectamineTM 48 h after transfection.

Effect of RRRαtocopherol Succinate on Antioxidant Capacity and Meat Quality in Broilers

ZHANG Xuhui;DU Haiming;ZHU Qiufeng;ZHOU Yanmin;WANG Tian

2010, 41(3): 286-294. doi:

Abstract ( 727 )

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The objective of this study was to compare the effect of two esters of αtocopherol, allracαtocopherol acetate (DLαTOA) and RRRαtocopherol succinate (DαTOS) on growth, antioxidant capacity and meat quality in broilers. 320 onedayold commercial Arbor Acres broilers were randomly distributed into 4 treatments, each of which had 8 pens of 10 chicks per pen. Birds were fed with the diets supplemented with 30 mg·kg-1 DLαTOA (control), the basal diet with DαTOS supplementation 10 mg·kg-1 (TOS1 group), 30 mg·kg-1 (TOS2 group) and 50 mg·kg-1 (TOS3 group), respectively for 42 d. The results showed that there was no significant difference (P>0.05) in the growth among the treatments. Significant positive correlations existed between dietary supplemental αTOS levels and plasma (R2=0.983 1, P<0.01) or hepatic (R2=0.933 6, P<0.05) αtocopherol concentrations, and a negative correlation with plasma (R2=-0.948 7, P<0.05) or hepatic (R2=-0.990 1, P=0.051 8) malondialdehyde (MDA) levels. In comparison with the control and αTOS1 groups, 3050 mg·kg-1 dietary αTOS supplementation resulted in an increase in the activities of serum TSOD, GSHPx, TAOC and hepatic TSOD, TAOC to some extent (P<0.05 or P<0.01). Furthermore, the level of hepatic ROS was decreased significantly (P<0.05). As for the meat quality, 48 h drip loss and shear force of breast and leg muscle was significantly decreased in broilers of TOS2 and TOS3 groups and also the cooking loss of leg muscle. But no significant differences were observed on pH45 min, pH24 h, 24 h drip loss and meat color (P>0.05). These results indicated that 3050 mg·kg-1 dietary DαTOS supplementation could enhance the antioxidant capacity of broilers, and further the water holding capacity and tenderness, which might result from increased retention of serum and hepatic αtocopherol and reduction in lipid peroxidation, as evidenced by the decrease in MDA and ROS.

Effect of Conjugated Linoleic Acid on the Composition of Myofiber Types in Skeletal Muscle Cells of Pigs in Vitro

HUANG Jinxiu;YANG Feiyun;LIU Zuohua;JIANG Shan;XIAO Rong

2010, 41(3): 295-300. doi:

Abstract ( 755 )

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This experiment was conducted to investigate the effect of CLA on the composition of myofiber types in skeletal muscle cells in vitro. The primary skeletal muscle satellite cells were used, and when the satellite cells were transformed into muscle cells, different CLA levels (0, 50, 100, 150, 200 μg·mL-1) were added into the cells. After D4, D8 and D12 of culture, the ratios of mRNA abundance of four isoforms of myosin heavy chain (MyHC I, MyHC 2a, MyHC 2b and MyHC 2x) were determined by semiquantitative RTPCR. The results showed that the composition of myofiber types in skeletal muscle cells changed markedly with culture time. From D4 to D12, MyHC 2b type fiber was increased markedly, but the other three types of muscle fibers were decreased significantly. The addition of 50 μg·mL-1 CLA had no effects on the composition of myofiber types. The effects of 100 μg·mL-1 CLA on the composition of myofiber types appeared mainly on D12, while 150～200 μg·mL-1 CLA affected significantly the composition of myofiber types from D4 to D12, with an upregulation of MyHC I and MyHC 2a type fibers, and a downregulation of MyHC 2x and MyHC 2b type fibers. The results suggested that the composition of myofiber types in skeletal muscle cells was influenced by CLA, which depended on its dose and culture time. The effect of CLA on the composition of myofiber mainly was represented by increasing the proportions of MyHC I and MyHC 2a type fibers but decreasing the ratios of MyH 2b and MyHC 2x fibers, which could explain partially why CLA improved pork quality.

Imunohistochemical Study on PrP Expression in Different issues of Tan Sheep in Ningxia

XU Lihua ;;KANG Xiaodong;TU Jian ;ZHANG Zhuming;YANG Jianmin;ZHOU Xiangmei;YIN Xiaomin;YANG Lifeng;ZHAO Deming

2010, 41(3): 301-304. doi:

Abstract ( 717 )

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In the present experiment, a qualitative detection was implemented to determine distribution and location of prion protein（PrP） in different tissues of Ningxia Tan sheep, which is one of the protected species of classⅠ. The result indicated that PrP has a wide range of expression in different tissues, such as cerebrum, brain stem, cerebellum, heart, liver, spleen, kidney, lymph node and so on. Especially, prion protein has a higher expression level in brain stem, spleen, kidney and lymph node than other tissues. We also got a negative result in the samples from lung. This study confirmed the status of distribution of PrP in different tissues from Tan sheep. In addition, the study provided a technical support to diagnose and detect scrapie by optimizing immunohistochemical method.

Identification of an Antigenicdrifted H5N1 Highly Pathogenic Virus in China

Yanbing;SHI Jianzhong;ZHAO Haidan;TIAN Guobin;WANG Dong;SONG Jiasheng;YANG Dequan;DENG Guohua;WANG Xiurong;CHEN Hualan

2010, 41(3): 305-309. doi:

Abstract ( 1193 )

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In order to evaluate an avian influenza virusCK/SX/2/06(H5N1), which was isolated from a poultry farm in Shanxi province. HAHI test, RTPCR, IVPI, antigenic analysis, sequencing and phylogenic analysis, aquatic bird and BALB/c mice infection test were fulfilled in the study. The virus were H5N1 subtype highly pathogenic avian influenza virus (HPAIV), which showed big diversity to reference strain GS/GD/1/96(H5N1) in China in phylogenic and antigenic analysis. So it was defined as antigenicdrifted strain, the same viruses established a new clade, Shanxi chickenlike strain, compared with the same subtype viruses which caused H5N1 HPAI outbreak in China in recent years. The virus didn′t infected SPF duck and showed low pathogenic to mammal. These results indicated that new clade and antigenicdrifted viruses Shanxilike strain appeared in China, so we should adjust the measurement and strategy to prevent and control the disease.

Comparision of Dynamic Detection with Three ELISA Kits to DifferentSubgroups of Exosomic ALVs in DF1 Cells

GUO Huijun;LI Zhongming;LI Hongmei;CHAI Jiaqian;MA Chengtai;WANG Hongjin;CUI Zhizhong

2010, 41(3): 310-314. doi:

Abstract ( 780 )

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DF1 cells were inoculated with three different doses (100, 10, 1 μL) of ALVA, ALVC, ALVJPY and ALVJWS separated from different chickens to evaluate three kinds of A, B, C ELISA kits. The samples of DF1 cells supernatant were obtained at different days post inoculation (dpi) and detected with three kinds of kits after freezing and thawing once. The results indicated that the samples inoculated with ALVA or ALVJPY were both detected positively firstly at the 3rd dpi with A ELISA kit for 100 μL inoculation doses and at the 7th dpi for 1 μL dose; If with B ELISA kit, two ALVs of subgroup were detected positively within some days post inoculation, but the detectable time were longer than that of A ELISA kit; If with C ELISA kit, the detectable time of viruses were the longest. ALVC and ALVJWS were detected positively with A ELISA kit at the 5th and 9th dpi for 100 μL doses, respectively; but for the other doses, no positive samples were detected at whole observed days. If with B and C ELISA kits, none of inoculated cells with three doses of inoculation was detected positively. It can be concluded that it is dramatically different between A, B and C ELISA kits in the detection of three exosomic subgroups of ALVs；The A ELISA kit is more sensitive than other two ELISA kits (B and C). Simultaneously, the positively detected days to ALVs are related to the doses of inoculation in DF1 cells. The results can be helpful for the application of three ELISA kits to the detection or isolation of exosomic ALVS.

Detection of the Interference to the Replication of Rabies Virus by BHK Cell Strains Transcribing shRNA Targeted to the N Gene

YANG Ruimei;;YANG Songtao ;WANG Chengyu;CUI Yan ;XIA Xianzhu

2010, 41(3): 315-321. doi:

Abstract ( 709 )

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In this study, the replication of rabies virus (RV) was inhibited by RNA interference in vitro experimentally in order to accumulate essential data of the application of RNAi in the researches of RV genomic function and the postexposure treatment of rabies. Targeting N gene of RV, four shRNA expression plasmids were designed and constructed based on the vector pRNATU6.3Hygro which expresses fusion protein of green fluorescent protein as a report gene. Four cell strains (BHKN1, BHKN2, BHKN3, BHKN4) expressing the short hairpin RNAs(shRNA) were obtained after the plasmids were transfected into the BHK21 cell line and screened under the pressure of Hygromycin B (300 μg·mL-1). These cell strains in a 24well tissue culture cluster were infected by 100TCID50 rabies virus CVS11 strain respectively, and 24, 48, 72, 96 h later, the virus replication were detected and evaluated by realtime RTPCR, 50% tissue culture infective dose(TCID50), directed immunofluorescence assay (DFA) and Western blot. At the 48th hour postinfection, the results of the realtime RTPCR indicated that four strains inhibited RV replication in various degree, in which BHKN2, BHKN1 had resulted in a better inhibitory effect(99%, 67.72%) than BHKN3, BHKN4(38.59%, 11.33%). TCID50 analysis showed that the virus titer of the cell strains were reduced by 24, 96.2, 2.14, 1.1 folds respectively compared with that of empty vector control. Inhibition ratios were 31%, 1%, 54%, 82% respectively compared with empty vector control detected by DFA. The Western blot results showed that the RV N protein bands were significantly weakened in BHKN2, BHKN1 cell strains. These findings suggested that the levels of replication of RV were dramatically decreased by BHKN1 and BHKN2 targeting the regions starting at positions 489 and 701 of rabies virus nucleoprotein gene. This study provided an alternative to block the infection of RV and to research the functions of RV genome.

Cloning and Sequence Analysis of Viable but Nonculturable State Related Genes in Escherichia coli

LI Ying;DUAN Rui;WANG Weili;SUN Xiaoyuan;QIAN Aidong

2010, 41(3): 322-328. doi:

Abstract ( 1036 )

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This experiment was conducted to study the molecular mechanisms of bacteria in viable but nonculturable (VBNC) state. Three differential fragments(A1, A2 and C1) were cloned from E. coli in VBNC induced by acetic acid and 4 ℃ with mRNA differential display PCR. These fragments were all extracted from E. coli in VBNC by reverse northern hybridization. The nucleotide homology between three sequences and E. coli 23S rRNA genes was 98%, 98% and 99%, respectively; and amino acids homology were all above 97%. These results indicated that normal E. coli without any selective pressure, its RNA transcript was lower, and some 23S rRNA genes were inhibited. But when in VBNC, some related genes expression were higher than that of normal state. These genes maybe participate in transcription or protein synthesis of E. coli in VBNC.

The Changes of Distribution and Structure of nNOS Positive Neurons in Rabbit Hippocampus after Ovariectomy and Estrogen Replacement Therapy

XIE Youlian;YIN Xunhe;QIU Jianhua;ZHAO Jun;WANG Xianlong

2010, 41(3): 329-334. doi:

Abstract ( 765 )

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To study the related changes of nitric oxide synthase (nNOS) positive neurons in the sections of the hippocampus of female rabbit (SpragueDawley strain) after ovariectomy and estrogen replacement therapy, the distribution and structure of nNOS positive neurons was observed in rabbit hippocampus using SABC immunohistochemical technique, which provides basic theories for preventing and treating Alzheimer′s disease with estrogen replacement therapy. The results showed: There are comprehensive nNOSpositive staining cells in all regions of hippocampus. After ovariectomy, the changes of nNOS positive neurons in subregion of hippocampus were not equal: the number of positive neurons in subiculum CA1, CA3 and dentate gyrus(DG) were decreased compared with control group (P<0.05), while the number of positive neurons in subiculum CA2 was increased (P<0.05). The average section area of positive neuron was smaller and the length of the longest process become shorter and the number of first tuber turns were less in subiculum CA1, CA3 and DG than that in control group(P<0.05). The average section area of positive neuron was smaller while the length of the longest process and the number of first tuber have no remarkable change in CA2 than that in control group (P>0.05). Four kinds index of positive cells in the subregion of hippocampus showed no significant difference between estradiolreplacement therapy and the control group(P>0.05).The results indicated that estrogen maybe affect the learning and memory by influencing the expression of nNOS in hippocampus.

Study on the Expression and Localization of βcatenin in Alpaca Skin of Different Colors

YU Xiuju;DONG Changsheng;FAN Kuohai;HE Junping;JIANG Junbing;ZHU Zhiwei;DONG Yanjun

2010, 41(3): 335-340. doi:

Abstract ( 782 )

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The objective of this study was to study the expression and localization of βcatenin in alpaca skin of different colors, and to explore the function of βcatenin in the process of coat color formation. The mRNA and protein expression level of βcatenin in alpaca skin of different colors were examined by Realtime quantitative PCR, Western blot and immunohistochemistry. The gene expressive quantity of βcatenin in the brown alpaca′s skin tissue was 1.662 times than ones of white alpaca. The distribution of βcatenin in alpaca skin were demonstrated，the expression was notable difference between the white and brown alpaca′s based on the average optical density. Our findings showed that βcatenin was involved in the process of coat color formation.

Localization and Development of Ghrelinimmunopositive Cells in Periphery Organs of Broiler Chickens

WEI Fengmei;LI Yugu;YE Yuanlan;ZHANG Yuan;MA Yongjiang;JIANG Qingyan

2010, 41(3): 341-346. doi:

Abstract ( 714 )

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In order to investigate the distribution of ghrelinimmunopositive cells in chickens, some samples of broiler chickens aged 2, 16, 30, 44 and 58 d were collected and studied by immunohistochemistry and microscopic image analysis. The ghrelinimmunopositive reaction was found in the glandular stomach, intestine, heart, liver, spleen, lung, kidney, pancreas, pituitary, adrenal gland, thymus, and bursa of Fabricius. The types of ghrelinimmunopositive cells (ghrelinic) were as follows: some endocrine cells and nerve plexes in the glandular stomach and intestine; Purkinje fibers in heart; Kupffer′s cells in liver; macrophages and reticular cells in spleen; pulmonary macrophages; podocytes, Intraglomerular mesangial cells, Juxtaglomerular cells in kidney; pancreatic islets cells in pancreas; some acidophilic cells and basophilic cells in pituitary; chromaffin cells in adrenal gland; thymic epithelial cells, thymic corpuscles and macrophages in thymus; some endocrine cells, reticular cells, macrophages and epithelial cells in bursa of Fabricius. With the chicken growth, the expressive contents of ghrelin were increased from aged 2 to 30 d, but slightly decreased at aged 44 and 58 d in various organs of chickens.

Effects of Apoptosis and NO Level of AVM on King Pigeon Brain Cells in Vitro Culture

LI Shu;CUI Yali;WANG Min;XU Shiwen;WANG Junjie

2010, 41(3): 347-352. doi:

Abstract ( 637 )

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The effects of apoptosis and NO level were explored with model of AVM on king pigeon brain neurocytes in vitro culture. Apoptosis was observed by TUNEL, the relative survival rate of neurocytes was detected by MTT, the percentage of apoptotic cells was detected by Flow cytometry, and the production of NO in cells was acquired by absorption spectrometry. The results showed that AVM can induce apoptosis and cytonecrosis of neurocyte, and has increased with the dose of AVM. AVM can reduced the production of NO, too, but no doseresponse relationship.

Effect and Mechanism of ACE2 on Injury Liver in Rats

LI Pengfei;ZHENG Junxi;ZHANG Wei;LI Lei;ZHANG Yuanshu

2010, 41(3): 353-359. doi:

Abstract ( 708 )

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The aim of this research was to observe the expression of ACE2 on injury liver which induced by CCl4, and its mechanism of antiinjury. 16 SD rats was randomly divided into two groups: control group and experiment group. The rats in control group were injected strokephysiological saline solution and the experiment group were injected 40% CCl4, respectively. The first dosage was 5 mL·kg-1, then 3 mL·kg-1 every three days. Four weeks later, the activities of ALT, AST and the concentrations of albumin, total protein , total bilirubin and AngⅡwere detected. We also analyzed the distribution and expression of ACE2 and Mas on liver by Realtime PCR and hybridization in situ. Compared with control group, the average daily weight gain, the concentration of albumin and total protein were markedly reduced (P<0.05), but the activities of ALT and AST were obviously increased (P<0.05).The concentrations of TBIL and AngⅡ had no significantly different compared with control group. The mRNA expression levels of ACE2 and Mas receptor were increased(3.21±0.52 vs 1.03±0.11, P<0.01;1.64±0.22 vs 1.02±0.10, P<0.05). The distribution of ACE2 was widespread throughout cirrhotic nodules, bile duct cells and endothelial cells lining small blood vessels. The expression of ACE2 and Mas receptor on injury induced by CCl4 in rats were increased significantly, which suggested that ACE2 may play an important role on injury liver by activating the ACE2Ang 17Mas axis to rival the effect of AngⅡ which could evoke injury.

Effect of the Disequilibrium of Calcium Homeostasis on the Apoptosis of the Neurons in the Chicken Brain Induced by Manganese in Vitro

CHEN Lei;LI Zhipeng;LI Shu;XU Shiwen

2010, 41(3): 360-365. doi:

Abstract ( 751 )

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To study the effect of the disequilibrium of calcium homeostasis on the apoptosis of the neurons in the chicken brain induced by manganese in vitro, neurons cultured in vitro were chosen as the research object. Neurons were cultured in the DMEM for 24 h with the final concentration of 0, 1.5, 2 and 2.5 mmol·L-1 MnCl2. The mitochondrial membrane potential (ΔΨm) and phosphatidylserine (PS) were assessed by flow cytometry. DNA fragmentations were detected by DNA Ladder. The calcium ion concentration within neurons (［Ca2+］i) was detected using the Fura3/AM as the probe and expression level of intracellular CaM mRNA was detected by FQPCR. The results showed that ΔΨm tended to decline the expansion of the pore, DNA fragmentations and ［Ca2+］i were increased. Moreover, the expression of CaM mRNA was decreased. This study indicated that MnCl2 could decrease the activity of CaM, ΔΨm and increase ［Ca2+］ result in apoptosis of the chicken neurons.

Effect of Ovulation Status on Efficiency of the Transplant in Recipient Gilts Transferred with Somatic Cloned Embryos

LUO Xueming;;ZHANG Weihong;;FENG Chong;LONG Chuan;YAN Jun;ZHAO Huijing;ZHANG Zhaowang;ZHANG Zijun;PAN Dengke;

2010, 41(3): 366-370. doi:

Abstract ( 806 )

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In this study, the effect of different ovulation status on pregnancy rates in surrogate recipient gilts receiving somatic nuclear transfer embryos was evaluated, in order to determine the optimal stage of the estrus cycle to achieve the highest pregnancy rates. Porcine somatic cell cloned embryos on 2 d in vitro were transplanted into the 12 d estrous recipient gilts. Recipient gilts were divided into two categories according to follicular development and ovulation status: just prior to ovulation or ovulating (Group 1); perovulation or after ovulation（Group 2）. The results were as follows：(1) pregnancy rate on 30 d of six recipient gilts from Group 1 was 100%; and the pregnancy rate on 30 d of ten recipient gilts from Group 2 was 40%, the data significantly different between Group 1 and Group 2 (P<0.05); (2) For the fullterm pregnancy rate of recipient gilts, extremely significant difference was observed between Group 1 and Group 2 (100% vs. 20%, P<0.01). These results suggested that development stage of porcine somatic cloning embryos on 2 d in vitro with recipient gilt′s womb environment of ovulation status of just prior to ovulation or ovulating was the best in the same period. It may achieve the best efficiency of embryo transfer.

Effects of Lutein on Reproductive Performance and Maternal IgG Transferring to Offspring of Quails

A Lun; DONG Xiaofang; TONG Jianming;ZHANG qi; WU Yingying

2010, 41(3): 371-376. doi:

Abstract ( 698 )

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This experiment was conducted to study the effects of lutein on reproductive performance and maternal IgG transferring to offspring of quails. 144 1dold parental generation North Korea Longcheng quails were randomly divided into four treatments groups with 3 replicates of 12 quails each. Wheatsoybean basal diet was used, and the treatment groups were supplied with 0, 50, 100 and 150 mg·kg-1 lutein. At 10 weeks old age, 2 quails each replicates were randomly selected and serum IgG levels were tested by ELISA. Then fertilized eggs were collected and hatched. Hatching rate, fertilization rate and death rate were calculated. Newly hatched birds were reared under the same condition, but without lutein in basal diet. On d 1, 3, 7, 14 and 21, 2 birds per replicate were selected and serum IgG levels were tested. The results showed that lutein could significantly improve fertilization rate and hatching rate of quails (P<0.05), significantly reduce death rate of embryos (P<0.05). Lutein had no significant impact on female quails serum IgG contents (P>0.05), but could significantly increase IgG deposition in egg yolk (P<0.05). Serum IgG levels of new hatched quails were correlated with lutein contents in the diet, and 150 mg·kg-1 lutein could significantly increase serum IgG levels of 1dold quails (P<0.05). Conclusion: Lutein could significantly improve reproductive performance of quails, and raise serum IgG level of offspring by promoting maternal IgG deposition in egg yolk.

The Truncated Mutation of Transmembrane Protein (TM or gp45) Identified in Fetal Donkey Dermal CellsAdapted Viral Strains (EIAVFDDV) of Chinese Equine Infectious Anemia Virus (EIAV) Attenuated Vaccine

MA Jian;SHI Nan ;LV Xiaoling;ZHAO Liping;JIANG Chenggang;LIN Yuezhi;KONG Xiangang;SHEN Rongxian;ZHOU Jianhua

2010, 41(3): 377-382. doi:

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To investigate the genomic evolution of EIAV vaccine strains during attenuation, we analyzed the sequence of env in provirus of EIAVFDDV. After amplifying env in provirus DNA of EIAVFDDV by using PCR, some positive TA clones were selected randomly to sequence and analyze. A mutation from TGG to TGA, a premature stop codon, was detected at the position of 2 128-2 130 nt of the env in 29 of 30 randomly selected clones of env. This mutation causes a truncated transmembrane protein (TM, also termed as gp45) at the 262th aa residue that resulted this mutated glycoprotein 154 residues shorter than the wild gp45. Indeed, we can see the band of predicted truncated gp45 when EIAVFDDV was analyzed by Western blot. To investigate the status of EIAV with the truncated gp45 in vivo, the env sequence of peripheral blood mononuclear cells（PBMC） associated provirus DNA and circulating virions in EIAVFDDVvaccinated horses were analyzed. TGG to TGA mutation were both found in integrated provirus DNA and the genome of the circulating virions. The above results indicate that the virion with the truncated gp45 owns the ability to infect the target cell and replicate in vivo. Extend studies are needed to understand the contribution of the truncated gp45 to immunogenicity and virulence of attenuated EIAV vaccine strains.

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