Acta Veterinaria et Zootechnica Sinica

Sequencing D-loop Region of Mitochondrial DNA in Yak and Study on Its Taxonomic Status in Bovinae

LI Qi-fa;LI Yin-xia;ZHAO Xing-bo;PAN Zeng-xiang;LIU Zhen-shan;ZHANG Qing-bo;QU Xu-Guang;SONG Da-wei;DONG Li-yan;LI Ning;XIE Zhuang

2008, 39(1): 1-6. doi:

Abstract ( 1586 )

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Primers designed according to the mitochondrial gene sequences of Bos taurus reported was used to amplify and sequence jiulong yak’s D-loop region, and the whole sequence was obtained. Using Ovis aries as outgroup taxa, the phylogeny about the representative species of Bovinae (Poephagus grunniens, Poephagus mutus, Bos taurus, Bos indicus, Bison bison, Bison bonasus, Bubalus bubalis）was analyzed. The results showed that the length of D-loop was 893 bp, owning 87.4% homology with the Bos taurus D-loop sequence, there existed 17 bp deficiency. Among Bovinae, the percentage nucleotide sequence divergence between Poephagus grunniens, Poephagus mutus and American bison of Bison bison was 6.2%-6.8%，which was less than that of Bos taurus and Bos indicus in Bos (10.0%-11.3%). Phylogeny analysis found that Poephagus grunniens, Poephagus mutus and Bison bison were clustered first of all, indicating there was higher genetic comparability among them than that of Bos. Combining with the proof of paleontology, morphology, molecular biology, the data supports the point that Poephagus grunniens and Poephagus mutus are classified as an alone genus in Bovinae, that is Poephagus.

Mitochondrial DNA D-loop Genetic Diversity and Origin of Some Chinese Domestic Buffalo Breeds

QI Guo-qiang;ZAN Lin-sen;ZHANG Gui-xiang;WANG Zhi-gang;WANG Jun-hui;HAN Xu

2008, 39(1): 7-11. doi:

Abstract ( 1269 )

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Analysis of the 110 mtDNA D-loop sequences (930 bp) in some 10 Chinese domestic buffalo breeds revealed that there were 50 different haplotypes and 107 polymorphic sites. The haplotype diversity(Hd) was 0.895 2±0.024 0, the nucleotide diversity(π) was 0.020 0±0.005 6, and the average number of nucleotide differences was 18.445 0. These results showed an abundant genetic diversity in Chinese domestic buffalo breeds. The NJ tree indicated that there were two main maternal origins in those 10 domestic buffalo breeds.

Cloning and PCR-RFLP Analysis of the Dopamine Receptor D4 Gene Partial Sequence of Horses

FAN Cai-yun;MANG Lai;CHENG Jian-bo;MORI Yu-ji

2008, 39(1): 12-15. doi:

Abstract ( 1257 )

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In this paper, partial sequence of horse DRD4 gene was cloned including intron1, partial exon1 and exon2. And restriction enzyme Stu I was used to analyze the polymorphism of the DRD4 gene sequences of 270 horses from 6 types including importing breed, cultivating breed and local breed. The products of digestion with restriction enzyme StuⅠ were detected by 8% nondenatured polyacrylamide gel electrophoresis and showed in silver staining protocol.The result indicated restriction enzyme Stu I showed polymorphism. Six kinds of genotypes were found in six populations , which were controlled by three alleles. The results of χ2 test showed that genotypes frequency of horse DRD4 gene in TB, SH, XN did not fit with Hardy-Weinberg equilibrium(P < 0.05), but that in WS, BH and WZ fit with Hardy-Weinberg equilibrium(P＞0.05) .

Studies on Microsatellite Markers of Fecundity Trait in Two Goat Breeds

SONG Yu-xuan;ZHU Guang-qin;WANG Yi-bing;WANG Jian-gang;CAO Bin-yun

2008, 39(1): 16-23. doi:

Abstract ( 878 )

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According to comparative genomics, four microsatellite loci including OarAE101, OarHH55, BM1329 and BMS2508 that linked to the fecundity gene FecB in the sheep chromosome 6 were analyzed for polymorphisms in Boer goat and Xinong Shaanen goat. The results indicate that there are genetic polymorphisms at these 4 microsatellite loci. The polymorphism information content (PIC) and number of effective alleles (Ne) are 0.834 0，0.722 2，0.820 1，0.685 5 and 6.719，4.098，5.610，3.542 respectively in Boer goats; the PIC and Ne are 0.765 5，0.738 7，0.670 1，0.682 0 and 4.849，4.360，3.520，3.609， respectively in Xinong Shaanen goats. The result of the effect of the 4 microsatellite loci on the litter size of 2 goat breeds showed that the allele 126 bp at OarAE101 has positive effects on the litter size in Boer goat but 134 bp has negative effects; the allele 120 bp and 124 bp at OarHH55 have positive effects on the litter size in Boer goat, but 130 bp/140 bp has negative effects; the allele 185 bp at BM1329 has positive effects on the litter size in Boer goat but 176 bp, 215 bp/240 bp have negative effects; the allele 118 bp at BMS2508 has positive effects on the litter size in Boer goat and 122 bp/141 bp, 148 bp has negative effects; the allele 108 bp, 114 bp/126 bp at OarAE101 have positive effects on the litter size in Xinong Shaanen goat and 124 bp has negative effects; the allele 120 bp at OarHH55 has positive effects on the litter size in Xinong Shaanen goat and 150 bp/165 bp has negative effects; the allele 185 bp at BM1329 has positive effects on the litter size in Xinong Shaanen goat and 176 bp, 215 bp/140 bp have negative effects; the allele 118 bp at BMS2508 has positive effects on the litter size in Xinong Shaanen goat and 141 bp, 153 bp/160 bp have negative effects.

Studies on the Differences of Phenotypic Values and mRNA Expression of Genes Associated with Fat Deposition in the Divergent-selected Chicken Lines

LI Dong-li;WEN Jie;ZHAO Gui-ping;ZHENG Mai-qing;CHEN Ji-lan

2008, 39(1): 24-28. doi:

Abstract ( 779 )

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Three groups of chickens were fed to study the differences of phenotypic values and mRNA expression of genes associated with fat deposition in the divergent-selected chicken lines. The IMF divergent-selected lines and unselected Beijing You chicken were fed and slaughtered at day 56 and 120. The phenotypic values of IMF content, abdominal fat weight, abdominal fat proportion and mRNA expression of LPL, H-FABP and A-FABP genes were measured and compared among groups. The results showed that IMF content and L* value of chicken muscle in upselected line (UL) was higher than the down-selected line (DL) at both day of 56 and 120, but the ultimate pH of breast muscle was lower. The differences were significant (P<0.05). At day of 120, the LPL in abdominal fat of UL expressed more than DL significantly (P<0.05), but the H-FABP in breast muscle expressed less significantly (P<0.05).

Regulation of Photoperiod on Seasonal Reproductive Activities and Endocrine in Magang Geese

HUANG Yun-mao;SHI Zhen-dan;LI Xiao-wei;LIU Zhi;LIU Ying

2008, 39(1): 29-36. doi:

Abstract ( 818 )

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To study the regulation of photoperiod on seasonal reproduction and the associated endocrine mechanisms in Magang geese, adult breeding geese and ganders were exposed to natural photoperiod (controls), or subjected to an artificial program of long and short photoperiods (experimental birds).In both groups, egg laying ceased after photoperiod increased, and onset and continued when photoperiod decreased or remained for 11 h.Treatment with the artificial photoprogram also increased the eggs laid by 85.6%, from the 26.3 eggs per control goose to 48.8 eggs on average in the experimental goose.In both control and experimental ganders, plasma LH concentrations were high in reproduction season from August to March or under short photoperiod, but decreased to low levels in non-reproduction season from April to July or under long photoperiod.A similar but much less marked seasonal pattern of plasma LH concentrations was observed in geese.In all geese and ganders, seasonal patterns of PRL concentrations were opposite to those of LH, in relation to changes of photoperiod.Plasma testosterone concentrations in ganders followed the seasonal secretion pattern of LH throughout experimental period.In addition, plasma T3 concentrations in both control and experimental ganders fluctuated throughout the experiment, but by no means correlated to changes of photoperiod or season.These results demonstrated that long photoperiod directly inhibited while short photoperiod directly stimulated reproductive activities in Magang geese and ganders.The photoperiodic effect was mediated via the secretions of PRL and LH, whose seasonal secretions coordinated the reproductive seasonality of Magang geese.

Plasma Gonadotrophin Concentrations during the Annual Cycle in Two Breeds of Goat with Low and High Fecundity

GAO Li-kun;GE Shi-hao;WANG Shu-ying;WANG Jian-min;HOU Yan-meng

2008, 39(1): 37-41. doi:

Abstract ( 740 )

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The concentrations of LH and FSH in blood plasma were measured by radioimmunoassay during the annual cycle of two local goat breeds in Shandong with low (Yimeng Black goat) and high (Jining Gray goat) fecundity. The results showed that the secretion of the two hormones in each breed changed with season obviously which was highest in autumn, secondary in spring and lowest in summer. FSH and LH levels in plasma showed a wave pattern in each breed during twelve months. The mean concentration of FSH was higher in Jining Gray goat than that in Yimeng Black goat in different month or season, but it was not significant (P>0.05 ). The level of LH in plasma was similar in the two breeds.

Regional and Ontogenetic Expression of NHE2 mRNA in Intestine of Broiler

WANG Xiu-qi;ZOU Shi-geng;ZUO Jian-jun;SU Hai-lin;HUANG Zhi-yi;FENG Ding-yuan

2008, 39(1): 42-47. doi:

Abstract ( 779 )

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120 1-day-old parental male Arbor Acre (AA) broilers were randomly divided into 4 replicates, respectively. Regional expression of sodium hydrogen exchanger 2 (NHE2) mRNA in different intestinal tract segments of AA broiler at 30-day-old and ontogenetic expression of NHE2 mRNA in duodenum and jejunum of AA broilers were determined by relative quantitative RT-PCR. It was found that: (1) Expression of NHE2 mRNA in duodenum, jejunum, ileum and colorectum was declined. The abundance of NHE2 mRNA in duodenum and jejunum was higher than that in ileum and colorectum (P<0.05)，respectively. (2)Ontogenetic patterns of NHE2 mRNA were same between duodenum and jejunum. The abundance of NHE2 mRNA was increased from 2- to 16-day-old, decreased from 30- to 44-old-day and increased again on 58-day-old. The abundance of NHE2 mRNA on 16- and 30-day-old were higher than that on 2-, 44- and 58-day-old (P<0.05)，respectively. These results indicate that regional expression of NHE2 mRNA in intestine of AA broilers was significantly declined along the downward intestine from proximal intestine to distal intestine (P<0.05). Ontogeny of NHE2 mRNA in AA broilers had the same pattern in duodenum and jejunum, indicating that NHE2 mRNA expression could be regulated by developmental stage.

Effect of Dexamethasone on Disacchridase Activities and Mucosa Morphology in Jejunum of Broilers

LI Yong;CAI Hui-yi;LIU Guo-hua;ZHENG Ai-juan;CHANG Wen-huan;ZHANG Shu;DONG Xiao-ling

2008, 39(1): 48-52. doi:

Abstract ( 717 )

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In order to explore the mechanism by which stress affects the growth of broilers, this trail was designed to investigate the effect of glucocorticoid, one hormone released when animal stressed, on disacchridase activities and mucosa morphology in jejunum of broilers. Thirty-six 21-day-old Arbor Acres broilers with similar weight was randomly assigned into four groups, three of which was injected with 1 mL dexamethasone-saline solution (containing dexamethasone 0.1, 1 and 5 mg/kg BW, respectively). The dexamethasone is a kind of drug like glucocorticoid. The control group was injected with 1 mL saline solution. The result showed that (1) The dexamethasone restrained the growth of broilers significantly, and the effect is depended on the dose of dexamethasone (P<0.05). (2) There is no significant differences found in both sucrase and maltase activities between control group and injected groups (P>0.05). (3) The dexamethasone decreased the villus height, the absorption area and the value of villus height/crypt depth in broilers' jejunum significantly (P<0.05). The crypt depth of the group injected with dexamethasone 5 mg/kg BW was significantly deeper than that of control group (P<0.05). But dexamethasone don't have significant effect on the villus width and lamina propria mucosae thickness(P>0.05). The results suggest that the reason of growth restrain of broilers caused by stress may be related to the secretion of glucocorticoid which gives rise to the decreases of villus height, absorption area and the increase of crypt depth , and then impairs the absorption function of intestine.

Serotype Identification and Virulenceassociated Genes Analysis of Pathogenic E.coli Isolated from Ducklings

YU Xue-hui;CHENG An-chun;WANG Ming-shu;WANG Ying;WANG Yuan-wei;TANG Cheng

2008, 39(1): 53-59. doi:

Abstract ( 938 )

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Among the 282 E.coli strains isolated from ducklings typically showing E.coli septicemia in large-scale duck farms in Southwest China, the serotypes of 210 strains were identified. The results showed that totally 37 serotypes were obtained, of which serotypes of O93, O78, O92 and O76 accounted for 43.8% and were the prevalent serotypes, and serotypes of O46, O32&O93, O60&O93 were isolated from ducks at the first time.The pesticin receptor gene (fyuA) within the highly pathogenecity islands (HPI), iron regulatory protein gene (irp), type 1 pilus essential protein gene (fimC), the pap pilus structural protein gene (papA) and serum tolerance associated gene (iss) of the 210 pathogenic E.coli strains isolated from ducks showing clinical signs and the 28 E.coli strains isolated from cloacal swab of clinically healthy ducklings were detected by the method of polymerase chain reaction (PCR) and nucleic acid sequence analysis.The results demonstrated that the genes of fyuA, irp2, fimC, papA and iss were detected from pathogenic E.coli strains isolated from ducks showing clinical signs and account for 41.0%, 43.3%, 92.9%, 97.6% and 96.7% respectively, and the corresponding genes detected from E.coli strains isolated from clinically healthy ducklings account for 21.4%, 25.0%, 92.9%, 100% and 92.9% respectively. The results showed that there were no significant difference (P＞0.05) of iss, fimC and papA gene between the E.coli strains isolated from ducklings showing clinical signs and that isolated from clinically healthy ducklings. With the respects of papA gene detection, there were significant difference (P<0.05) between the E.coli strains isolated from ducks showing clinical signs and that isolated from other hosts of chickens, swine and humans. The fyuA gene was widely distributed in ducklings and its detection rate in ducklings showing clinical signs was significantly higher (P<0.05) than that in clinically healthy duckling. There was a close positive correlation between the fyuA gene and the pathogenicity of E.coli. Furthermore, the fyuA gene was closely related with the specific serotype of O78. The detection rate of fyuA, irp2, fimC, iss and papA in ducklings showing clinical signs (37.6%, 79/210) was significantly higher (P＜0.01) than that in clinically healthy ducklings（14.3%, 4/28）.

Development of a Rapid Chromatographic Strip for the Diagnosis of Serotype O Foot-and-mouth Disease Virus

JIANG Tao;LIANG Zhong;CHEN Juan;HE Ji-jun;LU Lü;MA Wei-min;TIAN Hong;LIU Zai-xin;LIU Xiang-tao

2008, 39(1): 60-65. doi:

Abstract ( 783 )

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To develope a rapid and accurate test for detecting serotype O foot-and-mouth disease virus (FMDV)based on the principle of gold immunochromatographic assay(GICA),guinea pig and rabbit polyclonal antibody against FMDV serotype O were purified by DEAE-sephrose.The guinea pig anti (FMDV) serotype O-IgG was conjugated with colloidal gold ,which was called gold probes,and was dispensed on the glass fiber. The rabbit anti (FMDV) serotype O-IgG and the goat anti-guinea pig IgG polyclonal antibody were separately sprayed on the nitrocellulose membrane as the test line (T line) and the control line (C line). The rapid gold immunochromatographic strip was assembled by the different accessory in regular sequence. If there was serotype O FMDV in sample, gold probes on the glass fiber and rabbit anti (FMDV) serotype O-IgG formed a sandwich complex with serotype O FMDV.Within the test line, the sandwich complex was immobilized, furthermore, concentrated and appeared a distinct red color on the test line.In the clinic test, a total of 53 field samples were comparatively detected with both GICA and reverse indirect hemagglutination assay (RIHA), and the positive rate were 95.45% and 90.91%, respectively.The evaluation test proved GICA established in this study is simple, rapid, sensitive and specific.It is suitable for basic veterinary medical laboratories.

Experimental Reproduction of Postweaning Multisystemic Wasting Syndrome(PMWS) in Commercial Piglets with Porcine Circovirus 2

WANG Xian-wei;JIANG Ping;WEI Xian-kai;LI Guang-ming;LI Jun-xing;DONG Xin-tian;JIANG Wen-ming

2008, 39(1): 66-71. doi:

Abstract ( 913 )

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Postweaning multisystemic wasting syndrome (PMWS) is a disease of nursery and fattening pigs characterized by growth retardation, paleness of the skin and dyspnea. It has been demonstrated that PCV2 is the cause of PMWS. The expression of the disease is dependent on the co-infection with PCV2 and other factors or immunostimulation. In this study, 32-day-old postweaning piglets without antibody to PCV2 were inoculated intranasally with PCV2. The effect of immunostimulation on the induction of PMWS was investigated by immunization with keyhole limpet hemocyanin(KLH) emulsified in incomplete Freunds adjuvant. Sixteen piglets were distributed to four groups,each with four: control, PCV2, PCV2+KLH and KLH. The results showed that the 2 groups inoculated with PCV2 showed growth retardation. But the clinical signs of pigs in the group inoculated alone with PCV2 were not clearer than those in group with PCV2+KLH. Pigs inoculated with PCV2 and immunostimulated with KLH had increased rectal temperature, and one of these pigs developed wasting and died in day 12 after challenge. PCV2 viremia was found from 4 days after challenge to the end of the experiment. PCV2 nucleic acid was found in the lung and lymph node from the pigs of two groups inoculated with PCV2 by PCR. Meanwhile, the pigs in the groups inoculated with KLH alone or control had no clinical sign and pathogen. It confirmed that PCV2 is the primary viral infectious agent of PMWS.

Phylogenetic Relationship of Cysticercus cellulosae Isolates in Henan Based on cox Ι Gene

ZHAO Guang-hui;ZHANG Gai-ping;NING Chang-shen;WANG Xuan-nian;ZHANG Long-xian;JIAN Fu-chun;LU Kun

2008, 39(1): 72-78. doi:

Abstract ( 674 )

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344 bp partial sequences of cox Ι gene were obtained from ten Cysticercus cellulosae isolates derived from several areas of Henan Province by RT-PCR. PCR products were cloned into pMD19-T vector and recombinant plasmids were sequenced. Nucleotide sequences of partial coxⅠgene were aligned using the ClustalX 1.81, MP and NJ trees were constructed using the software PAUP version 4.0, and ML tree was also constructed using PUZZLE version 5.2. Sequence homology analysis was performed using the software WDANSIST version 2.5 and the Megalign program of the software DNAStar version 5.0. Results indicated that there was no sequence difference among the coxⅠ gene partial sequences of ten Cysticercus cellulosae isolates, which belonging to Asian genotype. cox Ι partial sequence of Cysticercus cellulosae can be helpful to distinguish validly between Asian and American/African genetype, and can be used for differential diagnosis of different Taeniidae tapeworm. Therefore, cox Ι is expected to be a differential marker for PCR differential diagnosis for taeniasis and cysticercosis.

Construction of Rabbit Immunity Model Against Haemaphysalis longicornis and Its Effects on H. longicornis Adult Growth

XIE Jun-ren;LIU Guang-yuan;TIAN Zhan-cheng;LI Zhi-xin;JIA Ning

2008, 39(1): 79-84. doi:

Abstract ( 1288 )

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In this study the rabbit immunity model against Haemaphysalis longicornis was constructed by infestation with H. longicornis adult quantitatively every two weeks. The results of ELISA showed that the antibody was positive three weeks post the first infection, the antibody level rising with the increasing of infection and reached the peak at the 11th week. The resistance of rabbit against H. longicornis significantly influenced the H. longicornis plump-blooding period. The antibody level was positively correlated to the bloodsucking period and the mortality of female H. longicornis adult and negatively correlated to the weight of plumpblooding female adult. These results indicated that the salivary gland is one of the main organs in the anti-H. longicornis immunity. The maintenance of the antibody level was long enough to make H. longicornis to complete a generation, which satisfied the experiment requirement. So it is an ideal model for anti-H. longicornis immunity positive control research. At the same time, the specific antibody level in the serum can be one of the important indexes to evaluate the extent of animal resistance to H. longicornis.

pound Mucosal Immune Adjuvants on Cellular Immune Responses of Small Intestine in Chicken

ZHANG Xiao-fei;YANG Qian

2008, 39(1): 85-90. doi:

Abstract ( 702 )

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The chicken were immunized orally with NDV strain LaSota vaccine containing two kinds of compound mucosal immune adjuvant(Group adjuvant Ⅰ and Ⅱ) respectively. The CD3+T lymphocytes, intraepithelial lymphocytes(IEL) and IFN-γ mRNA expression of small intestine in chicken were investigated. The results showed that the number of CD3+T lymphocytes and IELs increased during the period of immunity. The number of CD3+T lymphocytes of Group adjuvant Ⅱ were higher than that of Group adjuvant Ⅰ (P<0.05) in duodenum and significantly higher (P<0.01) in jejunum at the 3rd week and 5th week after vaccination. The number of IELs of Group adjuvant Ⅱ were significantly higher than that of Group adjuvant Ⅰ (P<0.01) in Peyer’s Patch at the 7th week after vaccination. The IFN-γ mRNA expression of two compound adjuvant groups decreased after vaccination. The results demonstrated that compound adjuvants could induce high level cellular immune response and promote intestine mucosal immunity in chicken.

Microstructure and Ultrastructure of Thyroid Gland in African Ostrich Chicks

LI Sheng-he;PENG Ke-mei;SONG Hui;WANG-Yan;WEILan;TANG Li;DU An-na;JIN Er-hui;WANG Jia-xiang

2008, 39(1): 91-96. doi:

Abstract ( 1115 )

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The microstructure and ultrastructure of thyroid gland of 2-month-old African ostrich chicks were observed by light microscope and transmission electron microscope technique. The results showed that the thyroid gland of ostrich chicks consisted of follicles and parafollicular cells. The follicles were different in size, and were mainly composed of simple cuboidal epithelial cells. The diameter of follicles ranged from 20.0 to 280.0 μm, and with an average of 69.9 μm. The height of follicular epithelial cells were from 5.0 to 15.0 μm, and were 9.4 μm on average. Under transmission electron microscope, the follicular epithelial cells could be functionally classified into type A and B. The shape of type A epithelial cell was cuboidal or low columnar, but that of type B epithelial cell was squamous or low cuboidal. The nucleus of type A was bigger and round or oviform, and the euchromatin in them was abundant, but the nucleus of type B was smaller and elliptic, and there was much heterochromatin in it. In cytoplasm of type A, there were a large number of RER, mitochondrion, Golgi complex and lysosome; but in the cytoplasm of type B, those organelles were underdeveloped. The parafollicular cells in the thyroid gland of ostrich chicks were oviform or polygonal in shape, lesser in number and bigger in volume. The parafollicular cells located among the follicular epithelial cells or between the follicles, and there were a number of RER, mitochondrion, Golgi complex and secretory granule in the cytoplasm. The above results suggested that the function of thyroid gland in 2-month-old ostrich chicks is active comparatively, could synthesize and secrete a large quantity of thyroid hormone and some certain quantity calcitonin.

Expression and Localization of Transforming Growth Factor-β1 in Alpaca Testis

YAN Yong-ping;DONG Chang-sheng;HE Jun-ping;Ren Yu-hong;HE Xiao-Yan;BAI Rui;Xie Jian-shan

2008, 39(1): 97-102. doi:

Abstract ( 790 )

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Transforming growth factor-β1（TGF-β1）plays important roles during testicular development and spermatogenesis，the main aim of the study is to determine the expression and localization of TGF-β1 in alpaca testis. The testes were obtained from alpacas at the age of 12- and 24-months（n=3）at the Scientific Research Base of Shanxi Agricultural University. The protein expression of TGF-β1 in alpaca testes was examined by Western blotting. The cellular localization and developmental changes were examined by immunohistochemistry SABC The immunostaining of TGF-β1 was present in leydig cells,sertoli cells,myoid cells and spermatogenic cells in the testis of alpacas at the age of 12- and 24-months，the optical density（OD）of TGF-β1 were respectively 0.730 5±0.011 5 and 0.562 6±0.012 2 in leydig cell；0.420 7±0.013 0 and 0.325 7±0.008 2 in sertoli cells；0.645 5±0.082 0 and 0.656 9±0.023 0 in myoid cells；0.457 9±0.009 2 and 0.374 3±0.015 8 in spermatogonia（P<0.01）. The positive immunoreactivity was located at the cytoplasm，not in the nucleus.No positive immunoreactivity was observed in the negative control. From the changes in pattern of localization of this peptide, we conclude that TGF-β1 is strictly regulated in leydig cells,sertoli cells and germ cells.It suggests that TGF-β1 involves in the regulation of spermatogenesis，which provides direct evidences for TGF-β1 action in the alpaca testis during postnatal development and spermatogenesis.

Telomerase Activity in Buffalo Oocytes and Early Embryos Derived from in vitro Fertilization

LI Xiu-lin;SHI De-shun;SHU Jin-hui;XIE Ti-san;CUI Kui-qing

2008, 39(1): 103-107. doi:

Abstract ( 790 )

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The objective of this study was to examine the telomerase activity in swamp buffalo oocytes and early embryos derived from in vitro fertilization (IVF). The telomerase activity of immature and matured oocytes, IVF embryos at the 2-4 , 8-16 cell, morula and blastocyst stages was assayed using a telomere repeat amplification protocol (TRAP). The telomerase activity was higher in the immature oocytes than that in the matured oocytes (P< 0.05). After in vitro fertilization, the telomerase activity decreased as the zygotes cleaved to the 2-4 cell and 8-16 cell stages, increased again when the embryos developed to the morula stage (P< 0.05), and reached the highest level at the blastocyst stage. When the telomerase activity was calculated on the individual cell, telomerase activity declined gradually as the immature oocytes matured to Metaphase II and developed to the blastocyst stage after in vitro fertilization. These results indicate that the changes of telomerase activity in the buffalo oocytes and early embryos are related to their maturation, embryonic developmental block and totipotency decline during the embryogenesis.

Stduy on Characteristics of Sodium-dependent Phosphate Intake in Small Intestine of Growing Pig

SUN Jie;ZHANG Tie-ying;CHEN Zhi-wei;HAN Jin-cheng;LIU Zhi-wei

2008, 39(1): 108-112. doi:

Abstract ( 1337 )

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Six 3-month-old growing Yorkshire pigs were killed to obtain mucosa, which used to prepare epithelium brush-border membrane vesicles (BBMV). The dependence of Pi uptake on pH and sodium concentration were discussed and kinetics of Na⁺-dependent Pi uptake into BBMV of growing pig were estimated by the measurement of phosphate uptake into BBMV within 15 s at room temperature. Results showed that acid pH made a negative effect on both sodium-dependent phosphate uptake and passive diffusion. Phosphate uptake into BBMV was strongly dependent on sodium concentration. Hill coefficient (napp) of sodium-dependent phosphate cotransport system was 2.07, which indicated that at least two sodium ion was transported with each phosphate ion. The differences of Pi uptake into BBMV between different intestinal segments were not significant. On the condition of varying Pi concentration (0-2.0 mmol/L) and pH7.4, the Pi concentration that yielded halfmaximal transport (apparent Km) estimated was (0.43±0.09)mmol/L in Yorkshire BBMV. The maximal phosphate uptake into BBMV was (2.25±0.19)nmol/(mg protein·15 s).

Study on Construction of Ovine Recombinant Plasmid pEGFP-PRNP and Its Expression in Eukaryotic Cell

HUO Gui-tao;ZHENG Jie;XU Guang-xian;YIN Xiao-min;ZHOU Xiang-mei;YANG Jian-min;ZHAO De-ming

2008, 39(1): 113-116. doi:

Abstract ( 696 )

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To construct a kind of recombinant plasmid with ovine prion protein gene （PRNP） and transfect the plasmid into C6 cell line in order to study the physiological function of normal prion protein and the pathogenesis in prion diseases. The gene sequence encoding ovine prion gene was amplified by PCR and cloned into eukaryotic expression vector pEGFP-N1 with EGFP reported gene encoding enhanced green florescence protein, and then identified by restriction enzyme. The recombinant plasmid pEGFP-PRNP was transfected into C6 cell line by using lipofectin method. We successfully construct the expression vector of recombinant plasmid pEGFP-N1 and it can be steadily expressed in form of fused protein in C6 cell line. It is suggested that the expression of recombinant plasmids pEGFP-PRNP in C6 cell line will provide a platform for researching expression of prion in cell and further contribute to study on prion diseases at cell level in future.

Preparation of Chicken SP-mRNA Probe and Its Detection in INR by in situ Hybridization Histochemistry

FENG Ya-mei;LIU Jin-xiong;LIU Yi;CHEN Qiu-sheng

2008, 39(1): 117-122. doi:

Abstract ( 723 )

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The fragment of SP gene was obtained by RT-PCR through total RNA of chicken brains. The amplified cDNA fragments were subcloned into pGM-T vector, and the plasmids were transformed into E. coli TOP10 and chosen by “white-blue plaque selection”. The recombinant plasmids were identified by sequencing. SP/pGM-T vectors were then linearized with the restriction enzyme of Nco Ⅰ and Sal Ⅰ, respectively. The sense and anti-sense DIG labeled RNA probes were producted by using SP6 and T7 RNA polymerase, respectively and in vitro transcription according to the protocol of “DIG RNA Labeling Kit (SP6/T7)”. Certificated by dot blot hybridization, the sense and antisense RNA probes were prepared successfully. The distribution of SP-mRNA in chicken INR was examined by using in situ hybridization histochemistry (ISHH). Most positive cells had oval and other polygonal shapes, and the labeled neurons distributed in ganglion in layers or small groups and in nerve trunk between ganglions sporadically. The proportion of positive cells in total neurons was 82.98% in the juxta-jejunoileum portion and 98.01% in the juxta-rectal portion of INR. In conclusion, the sense and anti-sense DIG labeled RNA probes for ISHH of SP were prepared successfully in this experiment which provided an approach for the further study of the location of SP-mRNA in nerve tissue. The results of ISHH suggest the existence of the SP-mRNA in most neurons of INR.

Mx Promoter-Luciferase Reporter Gene Assay for the Quantification of Chicken Type Ⅰ Interferon

CHEN Wei-ye;GE Jin-ying;ZHAO Hui-jun;HU Sen;WEN Zhi-yuan;CAO Wen-yan;WANG Xi-jun;HUANG Ke-he;BU Zhi-gao

2008, 39(1): 123-128. doi:

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In this study, the chicken Mx promoter（Mxp）was cloned, and luciferase (luc) gene was inserted into downstream of Mxp. Then pMxp-luc was constructed for the bioactivity assay of type Ⅰ IFN and the assay was developed and described as follows. Chicken embryo fibroblast cell line (DF-1) was transfected with pMxp-luc. 24 h later, the cells were incubated with 10-fold diluted chicken IFN-α and IFN-β for another 6 h. The expression of luciferase was stimulated by Mxp and had a good linear correlation with the antiviral activity of IFN. This new method was 10- and 100-fold more sensitive than the antiviral assay to quantify chicken IFN-α and IFN-β, respectively. In conclusion, the reporter gene assay described here is more rapid, safe and accurate than antiviral assay, and has potential value for high-flux assay and great value in the fundamental and application study of avian and mammalian type Ⅰ IFN.

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