Acta Veterinaria et Zootechnica Sinica

Current Progress in Copy Number Variation Study

LIU Xin;WANG Li-gang;WANG Li-xian

2010, 41(8): 927-931. doi:

Abstract ( 777 )

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Copy number variation (CNV) has recently gained considerable interest as a source of genetic variation likely to playing an important role in researching phenotypic diversity and evolution. CNV is a DNA segment ranging from kb to Mb. Much effort has been made in the identification and mapping of regions varying in copy numbers among normal individuals in human and a number of model organisms, using bioinformatics or hybridizationbased methods. The further researches have found that copy number variation is associated with mechanism of disease.In this paper, the outline, detection methods and research progress of the CNV are described and introduced, some existing problems in CNV research are presented, and the foregrounds of the research in human and animal diseases are prospected.

Genetic Monitoring on Genetic Diversity of Mashen Pig Using 21 Microsatellite Markers

CAO Guo-qing;LI Bu-gao;SHI Jian-zhong;LIU Hong;XUE Shang-jun;ZHOU Zhong-xiao

2010, 41(8): 932-938. doi:

Abstract ( 1061 )

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The aim of this study was to evaluate the conserved effect for Mashen pig by analyzing the trend of genetic diversity of Mashen pig populations during 1999 to 2007 and to make the effective measures for conservation. A total of 21 microsatellite markers recommended by FAO-ISAG were used to study the genetic diversity and the changing trend of Mashen pig. The results showed that there were 105 alleles detected in the overall Mashen pig population. The mean allele number per locus and mean effective allele number were 5 and 2.244 0, respectively. The mean observed heterozygosity and the mean PIC were 0.317 6 and 0.391 9, respectively. From 1999 to 2007, the mean allele number per locus, mean effective allele number, mean observed heterozygosity and the mean PIC of Mashen pig population declined from 4.285 7 to 3.428 6, 2.509 5 to 1.977 0, 0.311 6 to 0.298 1 and from 0.441 to 0.341, respectively. The results indicated that the homozygosity of Mashen pig was high and not polluted by other pig breeds, but the genetic diversity was becoming lower. The real-time monitoring on the change of genetic diversity of Mashen pig is needed in the future conversation and usage of Mashen pig. We should pay more attention to some alleles with low frequency to avoid them lost during generation passage.

Cloning of Goose Perilipin Gene, Tissues Expression and the Effect of Overfeeding on Its mRNA Level

PAN Zhi-xiong;WANG Ji-wen;TANG Hui;XIANG Shu-xia;WANG Jing;LV Jia;YANG Chao;HAN Chun-chun

2010, 41(8): 939-943. doi:

Abstract ( 1026 )

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This experiment was conducted to study the effect of overfeeding on Perilipin mRNA level in goose. In this study, the Landes goose and Sichuan White goose were used to clone the partial gene sequence of Perilipin, and the effect of overfeeding on the transcriptional level of Perilipin in liver was researched. The result indicated that the obtained Perilipin gene sequence was 277 bp, and had high similarity with other species. The mRNA expression of Perilipin gene was detected in different tissues of Sichuan White goose, and its expression level was higher in abdominal adipose and subcutaneous adipose tissues. Overfeeding markedly increased the mRNA expression of Perilipin gene in the liver of two breeds. In control group, the mRNA level of Perilipin in liver of Landes and Sichuan White goose was not different; compared with control group, in overfeeding group, the mRNA level of Perilipin in liver was markedly higher in Sichuan White goose and Landes goose, and there was significant difference at the mRNA level of Perilipin in liver between the two breeds. The mRNA abundance of Perilipin was positively correlated to the relative weight of liver, TG levels and total fatty acids in liver after overfeeding, and the correlation in Landes goose was stronger than that in Sichuan White goose. It was concluded that overfeeding induced the significant increase of Perilipin mRNA level in goose liver, and the effect of overfeeding was different between the two breeds.

Association of 14 Polymorphic Sites of the DRD2 Gene with Chicken Egg Laying Traits

XU Hai-ping;ZHOU Min;FANG Mei-xia;;ZENG Hua;NIE Qing-hua;ZHANG De-xiang;ZHANG Xi-quan

2010, 41(8): 944-950. doi:

Abstract ( 1084 )

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The purpose of this study was to analyze the associations of the DRD2 gene with chicken egg laying traits and find important markers for these traits. Fourteen polymorphic sites of this gene were selected, and the association analysis was performed in 644 chickens of Ningdu Sanhuang female line by PCR-RFLP and sequencing methods. The results showed that 4 mutation sites in the 5′regulatory region of the DRD2 gene, G-38560C, T-38326G, T-32751C and A-16105G, were significantly associated with the age of first egg (AFE) (P<0.05 or P<0.01). A-16105G was also significantly associated with total egg number at 300 days (EN300) (P<0.05). The haplotype block consisting of G-38560C, G-38544A, I-38463D and T-38326G was significantly associated with AFE (P<0.05). It was suggested that G-38560C, T-38326G, T-32751C and A-16105G on the DRD2 gene and the haplotype block consisting of G38560C, G-38544A, I-38463D and T-38326G could be used as effective molecular markers in marker assisted selection (MAS) for egg laying traits.

Association between Single Nucleotide Polymorphism of ActRIIB Gene andVertebra Number Variation in Small Tail Han Shee

LIU Jian-ming;SUN Shao-hua;HAN Li-xia;LI Xue-mei;SUN Zhi-ying

2010, 41(8): 951-954. doi:

Abstract ( 705 )

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This research was designed to detect the association between single nucleotide polymorphisms (SNPs) of ActRIIB gene and vertebra number variation in Small Tail Han sheep (Ovis aries). Using SNP analysis techniques, specific primers were designed to amplify the sequences of ActRIIB gene of the Small Tail Han sheep, PCR products from sheep of different thoracolumbar spine amount phenotypes were sequenced. A C85T mutation was found in the amplified fragment which caused MspⅠrestriction site disappear. Three genotypes, AA, AB and BB were produced after digestion by restriction enzyme MspⅠ, of which BB was mutation homozygote. The result of Chisquare test showed that genotype frequency was significantly different among different vertebral patterning sheep (P<0.05). The homozygous mutation was only found in normal T13L6 and T14L5 individuals, its frequency was highest in individuals with T14L5 phenotype. Thus it could be inferred that the mutation may give rise to the first lumbar vertebra changing into the fourteenth thoracic vertebra. At the same time, the effect of the point mutation in the 4th intron′s on the vertebra development of Small Tail Han sheep was found.

Study on Different Gene Expression in Follicular Granulose Cells of Huanghuai Goats with High Prolificacy

PANG Xun-sheng;WANG Zi-yu;YING Shi-jia;ZHANG Yan-li;WU Yong-cong;YAN Yi-bo;Meng Li;ZHONG Bu-shuai;WANG Feng

2010, 41(8): 955-961. doi:

Abstract ( 693 )

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The aim of this study was to analyze the gene expression during the goat antral follicle development. The different expressed genes in granulosa cells between non-latching large follicles (LF, 6 mm) and small follicles (SF, 4 mm) during follicular phase of Huanghuai goat ovary were studied using suppressive subtractive hybridization (SSH), and reduction cDNA library were also established. Furthermore, 96 clones randomly picked from the forward library were screened for differential expression by dot blot analysis. The results showed that eight clones had highly similar with the known genes, and twelve clones were new expressed sequence tags(ESTs). The results indicated that these genes may influence the maturation and ovulation of goat follicle.

Effect of Co-culture System with Vero Cells on the Development of Porcine Embryos

FU Bo;MA Hong;REN Liang;ZHAO Jin-feng;FANG Qing-chang;GUO Zhen-hua;LI Zhong-qiu;LIU Di;

2010, 41(8): 962-966. doi:

Abstract ( 639 )

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This study was conducted to investigate the effect of co-culture system with Vero cells on the development of porcine embryos. Ovaries were recovered from slaughterhouse, and oocytes were extracted. Oocytes were activated with electrical stimulation combined with chemical stimulation, then parthenogenetic embryos were obtained. Cloned embryos were constructed with porcine cumulus cells and oocytes. Cloned embryos and parthenogenetic embryos were co-cultured with Vero cells in NCSU-23(culture medium). The results showed that groups co-cultrued with Vero cells achieved significantly higher blastocyst formation rate compared with control(P<0.05), while there was no significant difference for the cell number of blastocysts(P>0.05). The result indicated that co-culture system with Vero cells improved the development of porcine embryos in vitro.

Effects of Growth-Related Genes Expression on Duodenal Developmentin Chinese Merino Sheep (Meat Line)

ZHOU Qi-wei;BAI Chun-sheng;JIA Bin;JIANG Wen-sheng;JIANG Song;DU Ying-chun;LI Ren-yan;WEN Qing-na;LV Li-min

2010, 41(8): 967-973. doi:

Abstract ( 945 )

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This experiment was conducted to study the effect of growth axis genes on growth performance of duodenum in Chinese Merino sheep(Xinjiang Junken type). Chinese Merino sheep were weighed and the length of duodenum was measured at 0, 7, 14, 30, 60 and 90 days. Meanwhile, pituitary gland, liver and duodenum samples were collected. Real time PCR was applied to investigate the developmental expression patterns of GHR, IGF-l and IGF-lR genes in liver and duodenum as well as GH and GHRHR genes in pituitary gland of Chinese Merino lamb (Xinjiang Junken type). Results showed that the expression patterns of GH and GHRHR genes in pituitary gland of lamb seemed to be similar to the expression of GHR genes in liver and duodenum, which increased and declined thereafter gradually. The developmental patterns of average daily length of duodenum gain and the expression pattern of IGF-1 mRNA were similar. These results indicated that the developmental regulation of duodenum was mainly affected by the expression of IGF-1 genes in liver and the expression of GH genes in pituitary gland.

Effect of Steam-Flaking or Extruded Corn and Full Fat Soybean in Starter on Ruminal VFA Ratio and Bacterial Ecosystem in Holstein Male Calves

ZHANG Yuan-qing;;MENG Qing-xiang;HE Dong-chang;YANG Xiao-min;WANG Fang;ZHANG Hong-gang;SHANGGUAN Ming-jun;ZHANG Bian-ying

2010, 41(8): 974-980. doi:

Abstract ( 990 )

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Experiment was conducted to investigate the effect of inclusion of different processing corn and full fat soybean in the starter diets on the rumen fermentation and the bacterial ecosystem in the rumen of Holstein calves. Holstein bull calves (n=12, (21±3) d) were blocked into 3 groups, and randomly assigned to receive a commercial pelleted starter containing either extruded corn and full fat soybean (PCON), steam-flaking corn and full fat soybean (SFCS), or ground corn and full fat soybeans (Control, NCON), respectively. Experiment lasted for 10 weeks from week 3 to 13 of age. Rumen fluid was collected for one or two weeks interval for the determination of rumen fermentation parameters and bacterial composition. During 10 weeks of the trial, total rumen VFA concentration tended to be influenced by different processing methods at week 6 and 9 (P>0.05). The molar proportion of butyrate for calves receiving SFCS starter diets was higher than those receiving PCON and NCON starter diets (P<0.05) at week 5 and 11. The bacterial composition analyzed by PCR-DGGE method showed similar profile among three groups but more complexes for SFCS group. All of the results indicated that the rumen fermentation development of calves in SFCS group was better than those in other groups.

Effects of Bee Pollen and Its Polysaccharides on Growth Performance,Nutrient Digestibility and Serum Biochemical Indexes in Calves

ZHANG Guo-feng;DIAO Qi-yu;TU Yan;YAN Ji-hong;ZHANG Nai-feng

2010, 41(8): 981-987. doi:

Abstract ( 818 )

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The experiment was conducted to investigate the effects of bee pollen (BP) and its polysaccharides (PS) on growth performance, nutrient digestibility and serum biochemical indexes in calves. 25 newborn Holstein calves were randomly allocated to 5 treatments with 5 calves each. A commercial milk replacer were fed to the calves. The five treatments were 0 g·d^-1(C), 10 g·d^-1 (10BP), 25 g·d^-1 (25BP), 50 g·d^-1 (50BP) of bee pollen and 5 g·d^-1 (5PS) of bee pollen polysaccharides substituted for milk replacer in equal amounts. The experiment started on 14 d after born and lasted for 56 days, using anus fecal method to determine the apparent digestibility of nutrient in two periods (21-28 d, 42-49 d).The results showed that. average daily gain of calves were significantly higher in 25BP and 5PS groups than that in C group（P<0.05). F/G decreased by 12.85% in 25BP group compared with that in C group（P<0.05);Apparent digestibility rate of dry matter in 21-28 d increased by 8.38% and 7.66% in 25BP and 5PS groups respectively compared with that in C group（P<0.05), meanwhile the apparent digestibility of crude protein increased by 18.03% in 25BP group（P<0.05); Serum TP and ALB content of calves fed bee pollen or polysaccharides were slightly increased, and BUN and CHO content were decreased. TG content were significantly decreased in treatment groups compared with that in C group(P<005). Supplement of bee pollen and polysaccharides in calves diet could improve the growth performance of calves, apparent digestibility rate of dry matter and crude protein, slightly increased serum TP and ALB content, reduced feed conversion ratio and serum triglyceride level significantly. Bee pollen additive in 25 g·d^-1 and polysaccharides in 5 g·d^-1 in milk replacer could get better performance and higher apparent digestibility of calves.

Exact Location of Linear B-cell Epitopes of VP3 Protein of Goose Parvovirus

GUO Lu;JU Huan-yu;YU Tian-fei;JING Zhi-qiang;MA Bo;WANG Jun-wei

2010, 41(8): 988-994. doi:

Abstract ( 689 )

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Four monoclonal antibodies against Goose Parvovirus (GPV) VP3 already obtained by our laboratory were used for the exact location to linear B-cell epitopes against VP3 of GPV. On the basis of the epitopes results of GPV VP3 protein that have already been identified in previous study, screened epitopes which can be identified by monoclonal antibodies directly. Oligomeric nucleic acid fragments of ten amino acid fragments mutually coinciding five amino acids were contrived, after annealing, connected with pET-32a, by allaxis identification, derivation expressed, small fragment fusion proteins were obtained. The antigenicity was identified by Western blot with McAbs. With the same method, amino acids were deleted one by one, and two epitopes were located at 430-435 aa and 643-647 aa.

Expression，Purification and Polyclonal Antibody Preparation and PreliminaryApplication of Phosphoprotein of Peste des Petits Ruminants Virus

ZHAI Jun-jun;DOU Yong-xi;ZHANG Hai-rui;MAO Li;MENG Xue-lian;WANG Qiu-xia;LUO Xue-nong;CAI Xue-peng

2010, 41(8): 995-1000. doi:

Abstract ( 692 )

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This experiment was conducted to express Peste des Petits Ruminants Virus (PPRV) phosphoprotein and prepare antiserum of phosphoprotein. For expressing the recombinant phosphoprotein in prokaryotic expression system, the recombinant plasmid pET-30a(+)-P sequenced correctly was transformed into Escherichia coli cells. Phosphoprotein was prokaryotic expressed in E. coli cell by induction of IPTG. SDS-PAGE analysis was performed to detect the recombinant protein. The highest soluble expression level of recombinant protein was obtained after being induced at 28 ℃ for 6 h and the concentration of IPTG was 0.2 mmol·μL^-1. The soluble recombinant protein was purified by Ni+ Sepharose affinity chromatograph method, and was subsequently used to immunize mouse. The anti-phosphoprotein polyclonal antibody with high sensitivity (1∶25 600) and specificity was obtained, and was used in detection of phosphoprotein eukaryotic expression in Vero. Phosphoprotein was highly expressed in E. coli, the antiserum of high titers and specificities was prepared, which lays the foundation for further research on phosphoprotein function.

Establishment and Clinical Application of a Multiplex Reverse TranscriptionPCR for Detection of Porcine Epidemic Diarrhea Virus, Porcine TransmissibleGastroenteritis Virus and Porcine Group A Rotavirus

ZHANG Kun;HE Qi-gai

2010, 41(8): 1001-1005. doi:

Abstract ( 1022 )

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To develop a multiplex reverse transcription polymerase chain reaction (mutiplex RT-PCR) for detection of porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV) and porcine group A rotavirus (GAR), three pairs of primers targeting the M gene, N gene ,VP7 gene of PEDV, TGEV, GAR were designed respectively. Using the three pairs of primes, a multiplex reverse transcription polymerase chain reaction (multiplex RT-PCR) was developed and optimized. And we compared the results of multiplex RT-PCR with routine RT-PCR in the field trail. In the laboratory test, we found that the detection limit of multiplex RTPCR is 35 pg RNA of TGEV-PEDV-GAR vaccine. And in the field trail, 75 fecal specimens collected from pigs with diarrhea in the central area of China were simultaneous tested by the multiplex RT-PCR and routine RT-PCR. The relative sensitivity and specificity of multiplex RTPCR were evaluated. The results suggested that this assay was equal in quality to routine RT-PCR assays (sensitivitys were 92%, 100%, 100% for PEDV, TGEV, GAR respectively; specificity was 100% for all 3 kind of viruses). The results indicated that the multiplex RT-PCR with high sensitivity and specificity provided a new and alternative tool for the detection of PEDV, TGEV and GAR.

Expression and Immunogenicity Study of Mevalonate Kinase Gene of Riemerella anatipestifer

LV Min-na;QIN Zong-hua;YUAN Jian-feng;SUN Ming-fei;YU Jin-shu;WU Cai-yan;CAI Jian-ping

2010, 41(8): 1006-1011. doi:

Abstract ( 643 )

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Mevalonate Kinase gene (RAMVA) was obtained when virulent genes of Riemerella anatipestifer were screened from DNA library by specific antibody as probes. The DNA fragment named DQ398751 had been record by GenBank. The length of open reading frame of RAMVA was 492 bp and encoding 163 amino acids. In order to express the gene, the digested PCR product of RAMVA was ligated with digested vector pMAL-C2X, and transformed into host bacteria E. coli BL21(DE3). Finally, the gene was successfully expressed in the bacteria by induction of IPTG, the content of the recombinant protein was about 15.6% of the whole bacteria proteins, and the recombinant mevalonate kinase was in a soluble state. Ducklings′ immune tests showed that two weeks post inoculation of the recombinant bacteria E. coli BL21/pMAL-C2X-RAMVA, the protection rate to Riemerella anatipestifer infection was 42.3%.

Establishment and Initial Utilization of a Duplex Real-time Fluorescence Quantitative PCR Assay for Detecting Brucella and Mycobacterium tuberculos

MENG Ru;CHEN Chuang-fu;ZHANG Hui;QIAO Jun;REN Yan;WANG Zheng-rong;JIANG Song

2010, 41(8): 1012-1017. doi:

Abstract ( 666 )

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To establish a method of duplex real-time fluorescence quantitative PCR assay for detecting Brucella and Mycobacterium tuberculosis simultaneously, we screened the optimimally specific primers of Brucella combined with the spesific cfp10 primers of Mycobacterium tuberculosi. Then the method was constructed and optimized. The specificity, sensitivity and reproducibility of this method were tested, and the clinic and imitation samples were detected. The results showed that the combination of omp25 primers of Brucella with cfp10 of Mycobacterium tuberculosis was optimun. The Tm of duplex RT-PCR to amplify Brucella and Mycobacterium tuberculosis was 88-89 ℃ and 90-91℃, and the result of amplification of other bacterias were negative. The lowest detection limit for DNA of Brucella, Mycobacterium tuberculosis or Brucella and Mycobacterium tuberculosis was 20, 50 and 100 copies·μL^-1, respectively, 100 times higher than conventional PCR. When the clinical and analogic samples were evaluated in parallel by this new assay and conventional PCR, the coincidental rate between the two tests was 100%. These results show that the assay is specific, sensitive and repetitive, and could be used in detecting Brucella and Mycobacterium tuberculosis simultaneously.

Expression of enJSRV in Immune Organs and Lung of the Fetus by In Situ Hybridization and RealTime PCR

WU Xiao-li;QI Jing-wei;LIU Shu-ying;XU Meng-jie

2010, 41(8): 1018-1023. doi:

Abstract ( 601 )

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To study the immunology mechanism of circulating antibody detected in sheep infected ISRV. Preparing the enJSRV-env probes, and we analysed the expression of enJSRV mRNA in immune organs (including thymus, spleen, gut lymph node) and lung of gestation 70 d fetal lambs, gestation 130 d fetal lambs and 7 d newborn lambs by In Situ Hybridization. The expression level of each target mRNA was analyzed in total RNA obtained from the above tissues by Real-Time PCR. The results revealed that enJSRV mRNA expressed in all collected tissues of different periods, while there were no positive signals in negative control. The results of Real-Time PCR showed that the level of enJSRV mRNA was much higher in immune organs than that in lung, especially in thymus and spleen of gestation 130 d fetal lambs and 7 d newborn lambs. This study will unravel the pathogenesis that sheep are tolerized towards infection by the exJSRV.

Localization, Distribution and Developmental Expression of Ghrelin in Thymus of BALB/c Mouse

LI Ying;LI Yu-gu;YE Yuan-lan;ZHANG Yuan;MA Yong-jiang

2010, 41(8): 1024-1030. doi:

Abstract ( 665 )

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The present study was conducted to examine the localization, distribution and developmental expression of Ghrelin in thymus of 1, 3, 5, 6, 12monthold BALB/c mice by immunohistochemical staining and microscopic image analysis. Ghrelin-immunopositive reaction was mainly distributed in the thymic medulla, less in the thymic cortex. The types of Ghrelin-immunopositive cells were subcapsular thymic epithelial cells, stellate thymic epithelial cells, medullary thymic epithelial cells, thymic corpuscle epithelial cells, macrophages and some lymphocytes in the thymic medulla, maybe thymic dendritic cells. The expressional contents of Ghrelin in thymus had not significant difference（P＞0.05） between 1- and 3-month-old mice, as well as 5- and 6-month-old mice, but that of 5- and 6-month-old mice were significantly lower（P＜0.05） than that of 1- and 3-month-old mice, as well as that of 12-month-old mice were significantly （P＜0.05）and extremely significantly lower（P＜0.01）than that of 5-, 6-, and 1-, 3-month-old mice respectively. The results suggested that the expressional contents of Ghrelin in thymus of mouse declined with age, and it might contribute to the ageassociated involution of thymus.

Effect of Dietary High Molybdenum on the Cell Cycle andApoptosis of Bursa of Fabricius in Broilers

Key Laboratory of Environmental Hazard and Animal Diseases of Sichuan Province;College of Veterinary Medicine;Sichuan Agricultural University;Ya′an 0;China

2010, 41(8): 1031-1038. doi:

Abstract ( 699 )

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The experiment was conducted with the objective of examining the effects of high molybdnum on the bursa of Fabricius in broilers by the methods of experimental pathology and flow cytometry (FCM).300 one-day-old Avian broilers were divided into four groups and fed on Control diet (Mo 13 mg·kg^-1) and High molybdenum diets(Mo 500 mg·kg^-1, High molybdenum groupⅠ; Mo 1 000 mg·kg^-1, High molybdenum groupⅡ; Mo 1 500 mg·kg^-1, High molybdenum group Ⅲ) for 6 weeks. The result showed that the weight and the relative weight of bursa of Fabricius were decreased, lymphocytes were histopathologically decreased and reticulocytes were increased in number in high molybdenum groupⅡand Ⅲ. Ultrastructurally, the frequency of lymphocyte apoptosis was higher in high molybdenum group Ⅱ and Ⅲ than that in control group. The mitochondria were swelled and mitochondria cristas were broken, or the density of mitochondrial matrix was increased. The statistical analyses by FCM indicated that the G0/G1 phase was increased (P<0.01) and the G2+M phase, S phase and the PI (Proliferating index) were decreased (P<0.01) of bursa in high molybdenum group Ⅱ and Ⅲ at 28 and 42 days of age. Meanwhile, the percentage of cellular apoptosis was higher in high molybdenum group Ⅱ and Ⅲ than that in control group. Also, the TUNEL staining was consistent with the result of FCM. It was concluded that dietary molybdenum in 1 000 and 1 500 mg·kg-1 inhibited the development of the bursa of Fabricius, caused pathological changes in the bursa of Fabricius in broilers.

Changes in Serum Fat Metabolism, Liver Function and Lipid Peroxidation of the Transition Cow

XIONG Gui-lin;FU Zhi-xin;CAO Sui-zhong;GU Jian-hong;CHEN Da-wei;LIU Zong-ping

2010, 41(8): 1039-1045. doi:

Abstract ( 1098 )

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Thirty Holstein dairy cows at 30 d before calving were selected for this study. Serum samples were collected at 30, 20, 10, 5 d before calving, 0 d calving and 5, 10, 15, 20 d after calving. Fat metabolism (concentrations of TG, Chol, NEFA, HDL and LDL), liver enzyme activities (AKP, LDH, ALT and AST activities), and antioxidation, lipid peroxidation (GSH-Px, SOD and CAT activities, MDA content) indexes in serum were measured. The result showed that: ① Concentrations of serum TG had a declining trend from 30 d before calving and decreased sharply on the day of calving. Concentration of Chol had a declining trend before calving and then increasing after calving. NEFA concentrations increased at calving, reaching peak level on day 5 after calving and started to decrease thereafter. Concentrations of serum HDL and LDL decreased slightly before calving period and increased gradually after calving. ② LDH and AST activities were the highest at day 5 before calving and the day of calving, respectively. ALT and AKP activities did not differ significantly at any stage. ③ CAT and GSH-Px activities showed a declining trend before calving period and increased at the day of calving followed by a decrease. SOD activity had increasing trend from day -30 to 0 and started to decrease thereafter. MDA content increased from day 5 before calving and decreased from day 5 after calving. In conclusion, there were the extensive mobilization of body fat and an imbalance in the oxidative status in the transition period of dairy cows.

The Study on Concentration of Endometrial Histamine and MastCell Graininess in Endometritis Cow

WANG Guo-qing;WANG Hong-hai;ZHANG Nai-sheng;WANG Hai-xia

2010, 41(8): 1046-1048. doi:

Abstract ( 672 )

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The study was focused on investigating the role of endometrial mast cells (MC) grains particle state and the histamine concentration in cow endometritis. Healthy and suffering from acute purulent endometritis Chinese Holstein cow were selected as control group and experimental group respectively, the number of each group was 10 and 6-10 days postnatal. The endometrial histamine concentration was detected by ELISA and the endometrial MC particle state was checked by transmission electron microscopy. The results showed that the concentration of endometrial histamine was (80.305±4.002) ng·mL^-1, (39.204±4.278) ng·mL^-1 in experimental group and in control group, the difference was more significant(P<0.01). In control group, inequality of size, homogeneous distribution, round/oval granule were discovered in MC cytoplasm of endometrium lamina propria; In experimental group, MC was degranulated, the quantity of granule decreased, the distribution was unevenness, cavity increment and vacuolization. Conclusion indicated that MC degranulation, released histamine in postpartum acute endometritis, leaded to the content of endometrial histamine significant increase, it concerned with endometrial acute inflammation, histamine could regulate the immunity function of uterus. It was very important to development of endometritis.

Development and Application of SYBR Green Ⅰ Real-time PCRTechnique for Detecting Porcine Astrovirus

SHANG Xiao-gui;GUO Wei;YANG Lian-ru;YANG Zhi-biao;HUA Xiu-guo;ZHU Jian-guo;CUI Li

2010, 41(8): 1049-1053. doi:

Abstract ( 592 )

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The aim of this study was to develop a SYBR Green Ⅰ real-time quantitative PCR for detecting porcine astrovirus(PASTV) quickly and flexibly. According to genome sequences within ORF2 of PASTV published in GenBank, a pair of primers was designed. The fragment of ORF2 gene was amplified with traditional PCR. The PCR product was cloned into pMD18-T vector and sequenced. The positive recombinant plasmid was used as quantitative template to generate standard curve and melt curve. Sensitivity assay, reproducibility of the assay and specificity assay were determined. The results demonstrated that standard curve established by recombinant plasmid shown a fine linear relationship between threshold cycle and template concentration. Melt curve was specific and the correlation coefficient was 0.994; the detection limit of real-time PCR for PASTV was 1×101 copies, and the quantitative PCR was more reproductive and specific than traditional PCR. These results indicated that the SYBR Green Ⅰ fluorescent quantitative PCR for detecting PASTV was developed for the basis of the early and rapid detection and analyzing the infect degree of PASTV quantitatively.

Identification and Phylogenetic Analysis of Newcastle Disease Virus Isolated from Duck Flocks

SUN Jie;DIAO You-xiang;LI Jian-xia;WU Huan-rong;YAN Yun;YANG Jian-peng;LI Yao-yao

2010, 41(8): 1054-1060. doi:

Abstract ( 701 )

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To explore the epidemiological understanding of Newcastle disease virus (NDV) in ducks, 10 NDVs isolated from outbreaks in duck flocks in Shandong Province during 2007-2009 were characterized both biologically and genetically. All 10 strains were velogenic strains based on their MDT (50.460.0) and ICPI (1.66-1.78). Ducklings challenged with the strains showed visible clinical symptoms and autopsy changes, and the morbidity and mortality were up to 70%-100% and 20%-70%, respectively. Compared with genotype Ⅰ-Ⅵ NDV, great variation were occurred, especially at 1-30aa, 101-124aa, 479-494aa and 509-516aa of F protein of the 10 isolates, which displayed the same characteristic amino acid substitutions at position 52, 71, 176, 272, 314, 402 and 489 with NDV strains obtained in recent years. Nucleotide analysis revealed that the homology among the 10 isolates was 95.8%-99.9%, and shared 95.7%-99.8% identities with genotype Ⅶ NDV since 2000, but only 83.8%-90.9% identities with LaSota and F48E9. Phylogenetic analysis showed that all the 10 isolates were of the genotype Ⅶd virus and had two RsaⅠ sites at position 973 and 1 249 bp which were characteristic to most strains of goose origin. The results indicated that the strains of genotype Ⅶd NDV had been the major pathogen responsible for the most epizootic ND outbreaks in duck flocks in China, and NDV isolated from ducks was likely originated from that of goose flocks.

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