Acta Veterinaria et Zootechnica Sinica

Expression Analysis of RPL29 Gene in Embryo Skeletal Muscle at Different Stages from Tongcheng and Landrace Pigs Using Realtime PCR Method

TANG Zhong-lin;DENG Hong;LI Yong;ZHANG Xing-ju;YANG Shu-lin;MU Yu-lian;CHU Ming-xing;MA Yue-hui;LI Kui

2008, 39(8): 1007-1012. doi:

Abstract ( 2097 )

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In order to investigate effect of RPL29 gene associated with protein synthesis on skeletal muscle development, the SYBR-Green quantitative real-time PCR (QPCR) method was established to analyze RPL29 gene. The expression change of RPL29 gene was analyzed in pig embryo skeletal muscle using the QPCR method. Based on standard samples with different concentrations, the standard curve “Y=-3.303X+43.077 (determination coefficient, R2=0.999)” was established. The results indicated the method had large test extension (Ct arranged from 13 to 35 cycle number), high amplification efficiency (100%) and single peak with unique PCR product in the melt curve. The Tm value was 86.5 ℃. In this study, the H3F3A gene was used for internal control, the expression change of RPL29 gene was analyzed in embryo skeletal muscle at 33, 65 and 90 days post coitus (dpc) from Tongcheng and Landrace using the QPCR method. We found that RPL29 exhibited the down- and up-regulation pattern in Tongcheng and Landrace pigs, respectively. There was higher expression in Tongcheng than that in Landrace at 33 dpc, and opposite result between two breeds at 90 dpc. No significant difference was found between two breeds at 65 dpc. These results indicated that RPL29 gene have potential association with skeletal muscle development and effect on meat production traits in pigs.

Optimization of Litter Size Model and Genetic Analysis of Model Parameters in Sows

ZHU Meng-jin;DING Jia-tong;LI Chang-chun;FAN Bin;YU Mei;LIU Bang;PENG Zhong-zhen;ZHAO Shu-hong

2008, 39(8): 1013-1018. doi:

Abstract ( 1586 )

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According to the phenotypic characteristics of life-cycle and fluctuation with parities, four mathematical models including Poisson cycle model, cubic polynomial model, modified quadratic polynomial model I and modified quadratic polynomial model II were respectively used to fit the expected litter size at different parities for Jiangquhai sows. From the viewpoints of statistics and biological meaning, modified quadratic polynomial model I was the optimum model of litter size. Single trait animal model and DFREML procedures were involved to estimate the heritabilities for optimum model parameters. The results showed that the values of heritabilities for model coefficients A and B and the estimation of acmes of pure quadric curve in model were higher than that for litter size, which hinted that selection on model parameters should be more effective than direct selection on litter size.

Microsatellite Markers and Association with Fecundity Trait in Boer Goat

XU Yun-xiang;CHEN Shi-lin;WEN Qun-ying;SHEN Zhong;ZHANG Chun-yan;YAO Hong-wei;HUA Guo-hua;YANG Li-guo

2008, 39(8): 1019-1024. doi:

Abstract ( 999 )

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According to the goat genetic linkage map of chromosome 2 and 8, eight microsatellite markers （INRA040, LSCV22, LSCV37, IDVGA64, FCB011, INRA129, SRCRSP10 and CSSM47） were chosen to analyze their correlation with the litter size of Boer goat. All the eight microsatellite loci in Boer goat were high polymorphism locus (PIC > 0.5). Ten alleles were found to be significantly correlated with litter size of Boer goat. 189 bp of LSCV22 weas positively correlated to first parity litter size of Boer goat. 179 and 191 bp of LSCV22, 273 and 303 bp of SRCRSP10 had negative correlation with first parity litter size. 189 bp of LSCV22, 281,283 and 245 bp of IDVGA64, 143 and 165 bp of FCB011 were positively correlated to second parity litter size.

Genetic and Statistical Analysis between Production Traits and Secondary Traits in Chinese Simmental

FAN Da-you;XU Shang-zhong;LI Jun-ya;REN Hong-yan;YANG Xue-li

2008, 39(8): 1025-1032. doi:

Abstract ( 1614 )

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Heritability and genetic correlations between 19 traits were estimated from data obtained from 6 Chinese Simmental farms using an animal model and MT-DFREML.Data relating to secondary traits and production traits in 1 634 Chinese Simmental cows were collected from 1980—2000.The traits analyzed were calving interval, open days, gestation length, milk yield, fat yield, protein yield, fat percentage, protein percentage, somatic cell score, milking speed, herd life, and linear type score (which included type score, rear leg view, fore udder attachment, rear udder width, rear udder height, teat placement, udder conformation, and udder vein). Estimated heritability of each trait were 0.08, 0.07, 0.08, 0.16, 0.15, 0.41, 0.11, 0.07, 0.10, 0.23, 0.18, 0.42, 0.13, 0.25, 0.35, 0.32, 0.23, 0.20 and 0.21, respectively. Genetic correlation between milking speed and production traits were negative, ranging from -0.22 to -0.10. Genetic correlation between herd life and type score was 0.87 and herd life and milking speed was 0.61. Genetic correlations between production traits and herd life and production traits and somatic cell score were unfavorable. Genetic correlations between herd life and milking speed, herd life and type score, herd life and milk yield, and herd life and somatic cell scores were 0.61,0.14,0.42 and -0.23, respectively.Udder traits were highly correlated with production traits and scondary traits. Where the highly correlated traits were fore udder attachment, rear udder width, teat displacement and udder vein, the genetic correlations ranged from -0.36 to 1.00. indicating that the production traits and secondary traits will be improved by selecting udder traits. Environment has a great effect on secondary traits, the management and milking stage were the main factors to influence milking speed. herd life and somatic cell count.

Cloning, Sequencing and Distribution of Chick β-defensins Genes

MA De-ying;LIU Sheng-wang;LI Yi-jing;SHAN An-shan

2008, 39(8): 1033-1039. doi:

Abstract ( 1711 )

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Three Gallinacin (Gal) genes: Gal-6, Gal-8 and Gal-9 were cloned from liver and tongue of chicks by RT-PCR, respectively. In addition, we have sequenced these genes. The full cDNA length of Gal-6 and Gal-9 consist of 201 bp encoding 67 amino acids, while Gal-8 consist of 204 bp encoding 68 amino acids. Differential gene expression of Gal-6, Gal-9 and Gal-8 has been demonstrated across a panel of tissues in the chick. It has been shown that Gal-6 mRNA was highly expressed in the skin, tongue, liver, heart, small intestine, breast muscle, kidney, bursa of Fabricius, spleen, and pancreas, and moderately expressed in the lung, testis, and borrow marrow. Gal-8 mRNA was highly expressed in the liver, tongue, small intestine and lung of chick. While, Gal-9 mRNA was highly expressed in the tongue and small intestine, and moderately expressed in the trachea, lung, liver, kidney, borrow marrow and thymus of chick.

FSH Regulating the Expression of GDNF of Piglet Testis Sertoli Cellby Activing ERK1/2

SUN Yan;WANG Xian-zhong;WU Jian-yun;BAI Ru-lan;ZHANG Jia-hua;

2008, 39(8): 1040-1044. doi:

Abstract ( 888 )

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Primary piglet sertoli cell (SC) were used to study the effects of FSH on regulating Glial cell linederived neurotrophic factor (GDNF) expression. The results showed that: (1) The FSH-stimulated GDNF expression in the cultured SC was time- and concentration-dependent: the level of GDNF protein reached its highest when FSH (50 ng/mL) stimulated 1h. (2) FSH（50 ng/mL）or dbcAMP（100 μmol/L）could active ERK1/2 prominently along with a maximum stimulation at 2 h; (3) U0126 (MEK1/2 inhibitor) could apparently inhibited the activing effect of FSH on GDNF. These results suggested that, by activing the cAMPPKAERK1/2 and promting the transcription of GDNF gene, FSH stimulated the expression of GDNF of sertoli cells.

Studies on in vitro Fertilization of Porcine Frozen Semen

DING Yi;;DAI Jian-jun;WU Hua-li;ZHANG De-fu;XU Gang-yi

2008, 39(8): 1045-1049. doi:

Abstract ( 1429 )

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In order to simplify the source of semen of in vitro fertilization(IVF), and avoid the effects by different boars and different semen collection time, we used frozen semen by 5 mL-straw to do the IVF. The results showed that there were no significant differences in the polyspermy rates among the 2-8 h oocyte-sperm incubation time(P>0.05). The 2-cell rate was improved along with the increasing of oocyte-sperm incubation time, and 8 h group (72.8%) was much higher than the 2 h group(60.5%, P<0.05). The morula and blastocysts rates of the 6 h group were the highest and got 46.1% and 3.48%, respectively. Polyspermy rate was also increased with the increasing of sperm concentration in the IVF (P<0.05). The 2-cell rate of 108 group(81.2%) was the highest, which was much higher than the 105 group. The 107 and 106 group got 8.11% and 3.13% blastocyst rate, respectively. Using NCSU-23 as the in vitro maturation(IVM) medium and matured oocytes partly denuded cumulus cells could get the highest cleavage and blastocyst rates(77.1%,7.33%) among the four groups. The comparation between the frozen semen and fresh semen in the IVF showed that there were no significant differences in the cleavage, morula and blastocyst rates(P>0.05).

The Effect of Feed Style on the Lipid Metabolism of Hetian Chickens

YANG Ye;WEN Jie;CHEN Ji-lan;ZHAO Gui-ping;FANG Gui-you;FENGYu-lan;LI Zhongrong

2008, 39(8): 1050-1055. doi:

Abstract ( 997 )

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The experiment was conducted to study the effect of feed style, sex and age on the lipid metabolism of Hetian chickens. The 28dHetian chickens were raised in three kinds of feed styles. The one reared in cagedfeed for 4 pens of each 30 chickens（♀15，♂15）. The other reared indoor floor for 4 floors of each 60 chickens(♀30，♂30). The third reared outdoor access for 4 pasture of each 100 chickens（♀50，♂50）. The male and female were fed separately. The chickens were slaughtered and analyzed at 56, 84 and 112 d, respectively. The results showed that the feed style , sex and age influenced significantly the lipogenesis FAS and ME mRNA levels and enzyme activity. The hepatic FAS and ME mRNA level and enzyme activity of freeranged chickens were lower significantly than that of caged chickens(P<0.05). Then the intramuscular fat(IMF), intermuscular fat(IeMF), subcutaneous fat pad(SFP) and percent of abdominal fat(PAF) levels of freeranged chickens were lower significantly than that of caged chickens(P<0.05). The hepatic FAS and ME mRNA levels increased with the age increasing. The female hepatic FAS and ME mRNA levels were higher than the male. The correlation between enzyme activity and lipid deposition indicated that the hepatic fatty acid synthase (FAS) activity was positively related with thigh intramuscular fat (T.IMF) ,IeMF, SFP and PAF and negatively with breast intramuscular fat(B.IMF). The hepatic FAS activity was significantly correlated with SFP and PAF(P<0.05). The hepatic ME activity was significantly positively correlated with B.IMF and T.IMF(P<0.05), and negatively with SFP and PAF(P>0.05).

Effect of Nutritional Level on PRKAG3 Gene Expression and Pork Quality

LI Meng-yun;YU Bing;ZHANG Ke-ying;CHEN Dai-wen

2008, 39(8): 1056-1061. doi:

Abstract ( 990 )

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12 DLY pigs(70 kg) were randomly divided into two groups with diets of high (DE 13.81 MJ/kg, CP 14%) and low (DE 12.55 MJ/kg, CP 11%) nutritional level,respectively. All pigs were slaughtered when their weight are about 100 kg, and determined PRKAG3 gene expression, carcass characteristics and meat quality traits. The results showed that low nutritional level increased PRKAG3 gene expression(P>0.05).High nutritional level increased dressing percentage, lean percentage, loin muscle area, a, b and L values（P>0.05）, but drip loss was significantly lower than that of low nutritional level(P<0.05). PRKAG3 gene expression was positive relative to lean percentage, loin muscle area, a value, b value and drip loss（P>005）, but significantly negative relation to pH2(P<0.05). These results suggested that nutritional level influenced PRKAG3 gene expression and so influenced meat quality.

Effect of Supplement Bacillus subtilis Natto on Ruminal Microbial Fermentation in vitro

DENG Lu-fang;WANG Jia-qi;JIANG Yan-mei;LIU Liang;BU Deng-pan;ZHOU Ling-yun

2008, 39(8): 1062-1068. doi:

Abstract ( 1078 )

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The purpose of the present study was to examine the effect of different concentration of Bacillus subtilis natto on ruminal microbial fermentation in persistent artificial rumen system. Three groups, including control group (CK), treatment 1 group (TR1) supplied 1% B.subtilis natto of fermenter capacity and treatment 2 group (TR2) supplied 5% B.subtilis natto of fermenter capacity were employed. Results indicated that， compared with CK, pH values of TR1 and TR2 increased, but the difference was not significant (P>0.05). The NH₃-N concentration of TR2 was always higher than that of CK (P<0.05), while TR1 failed to increase NH₃-N concentration significantly (P>0.05). Compared with CK, the MCP concentration of TR1 and TR2 decreased significantly (P<0.05) and their concentration of acetate, propionate, butyrate, and total volatile fatty acids all increased to different extent. Compared with CK, the concentration of acetate, propionate, butyrate, and total volatile fatty acids in TR2 increased by 6.52%, 1.96%, 11.02% and 5.77% (P<0.05), respectively. TR2’s acetate/propionate ratio also increased significantly (P<0.05). However, no significant difference was observed between TR1 and TR2 (P>0.05) in all volatile fatty acids except that butyrate concentration was higher in TR2 than that in TR1 (P<0.05). The results suggested that B. subtilis natto has a negative effect on N metabolism, but could enhance the carbohydrate digestion by rumen microbes.

Digestion of Diets for Early Weaning Lambs and Their Effects on Development of Lamb Alimentary Tract

GUO Jiang-peng;;WANG Hong-bo;LI Fa-di;HAO Zheng-li

2008, 39(8): 1069-1074. doi:

Abstract ( 1654 )

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The trial was combined with a feeding experiment, in which twentyfour three-way cross-bred lambs(Poll Dorset♂×Small Tailed Han×Tan♀, averaged at 35.5 days old) were divided into three groups and each with eight lambs(♂1,♀7)，pen-fed singly. The experiment lasted sixty-five days including 15 days for the preliminary period. Lambs were fed with three diets at different levels of digestible energy(DE) and crude protein (CP),i.e.I- 0.9NRC,II- 1.0NRC and Ⅲ-1.1NRC， respectively, during the recording period of 50 days.Sampling of the digestion trial using five female lambs for each group were made from the days 21 to 26 during the recording period, and four lambs(♀) in each group were slaughtered at the end of the feeding experiment to study the digestion of these diets and the development of alimentary tract for lambs. The results showed that the apparent digestibility of dry matter, organic matter, crude protein and acid detergent fiber（DMD, OMD, CPD,ADFD） increased with the raising of nutrition level in diets. DMD and OMD were higher for diet Ⅲ than that for Ⅰ（P＜0.01）and Ⅱ（P＜0.05）; CPD from Ⅲ was higher than that from Ⅰ and Ⅱ（P＜0.01）, DMD of Ⅱ was higher than that of Ⅰ（P＜0.05）.The ADFD of Ⅰ was lower than that of Ⅲ（P＜0.01）. The changes of apparent digestibility of calcium and phosphorus (CaD ,PD) were not regular as nutrition levels. The weight of alimentary tract(% of lifeweight before slaughtering) and the weight of each compartment of stomach(% weight of whole stomach) were not affected by nutrition levels（P＞0.05）. The weight of rumenreticulum(% weight of whole stomach) for the lambs of all groups had reached the levels of adult sheep.

Appearance of bla_CMY-2-producing Escherichia coli Isolates of Chicken Origin in Shandong Province and Studies on Their Relevant Characterizations

ZHANG Yong-ning;LIU Hong-jie;NIU Zhong-xiang

2008, 39(8): 1075-1080. doi:

Abstract ( 1419 )

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126 E. coli isolates of chicken origin were collected from different areas in Shandong province. With the specific primers for the blaCMY-2 gene, 15 isolates were demonstrated to be positive for blaCMY-2 by PCR amplification. Nucleotide sequence analysis demonstrated that the blaCMY-2 sequences of these isolates were identical to that published from GenBank except for strain JN10. Susceptibilities of the 15 isolates to the selected antimicrobial agents were determined by the Kirby-Bauer disk diffusion method, which demonstrated that all the 15 isolates were resistant to cefalothin, cefazolin, cefaclor, cefixime, ampicillin, streptomycin, tetracycline, chloramphenicol and trimethoprim/sulfamethoxazole. Three (JN5, JN9, and HZ9) of the 15 isolates were also resistant to ceftriaxone and ceftazidime. But all isolates were sensitive to cefepime. The modified three dimensional test demonstrated that all the 15 isolates produced CMY-2-type AmpC β-lactamases and four of the 15 isolates produced extendedspectrum βlactamases (ESBLs) at the same time. In addition, the plasmids harbouring the bla_CMY-2 gene were demonstrated to be successfully transferred from E.coli isolates to the recipient E.coli DH5α (sodiumazide resistant mutation) by conjugation experiment. The possibility of the 15 isolates deriving from the same clone was excluded by pulsed-field gel electrophoresis. This study provides a very important theory and research foundation for probing into and resolving the problems of drug-resistance among bacteria. Besides, it also provides important surveillance information for knowing the drug-resistance condition of these E.coli isolates and their dissemination in Shandong province.

Studies on an Immunochromatographic Method for Rapid Detection of H9 Subtype Avian Influenza Viruses

PENG Fu-hu;WANG Zheng;ZHANG Jin-liang;ZHANG Wen-tong;ZHANG Zhan-ying;HU Si-shun;XIAO Yun-cai;BI Ding-ren

2008, 39(8): 1081-1086. doi:

Abstract ( 1412 )

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An immunochromatographic strip was developed for the detection of the H9 subtype of avian influenza viruses (H9AIVs) in poultry using McAb 4C4 for H9AIV hemagglutinin and chicken anti-AIV antibody. The 4C4 was labeled with colloidal gold as a detection reagent, and chicken anti-AIV antibody was blotted on the test line while a goat anti-mouse antibody was used on the control line of the nitrocellulose membrane. In comparison with the hemagglutination (HA) and hemagglutination inhibition (HI) test, the strip was specific for detecting H9AIV with the sensitivity at 0.25 HA units within 10 min. Storage of the strips at room temperature for six months or at 4 ℃ for 12 months had not changed their sensitivity and specificity. 311 tracheal samples from potentially infected chickens were detected using the strip and ELISA kit established in our lab, the coincidence ratio was 92%. Further characterization of 10 positive samples randomly selected showed that no single sample was false positive, as determined by the HI assays and standard virus isolation. Our research revealed that the immunochromatographic strip for detection of H9AIVs has high specificity, sensitivity and stability. This, together with the advantages of rapid detection and easy operation without requirement for special skills and equipments, makes it suitable for the on-site detection and differentiation of H9AIVs from other viruses in poultry.

Clone，Prokaryotic Expression of DPV dUTPase Gene and Its Subcellular Localization in Virus-infected Host Cells

ZHAO Li-chan;CHENG An-chun;WANG Ming-shu;YUAN Gui-ping;JIA Ren-yong;ZHOU Deng-chun;GE Han;SUN Tao;CHEN Xiao-yue

2008, 39(8): 1087-1093. doi:

Abstract ( 949 )

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A pair of primers was designed based on the DPV dUTPase gene sequence that our laboratory newly discovered (GenBank accession number DQ486149). Amplified PCR fragments were subcloned into the prokaryotic expression vector pET-32a (+). Then the recombinant plasmid pET32a-DU was transformed into E.coli BL21（DE3）strain and expressed under IPTG induction. SDS-PAGE analysis showed that the induced expressed protein was about 66.8 ku and accounted for 36.2% of total bacterial protein by gel scanning. The protein was purified and used to immunize rabbit for the dUTPase anti-serum production. Its ELISA antibody titer was up to 1∶409 600.Moreover, the subcellular localization detection was observed using immunofluorescence technique. The results showed that specific fluorescence appeared in cytoplasm as early as 4 hours post infection and a great deal of specific fluorescence concentrated in the cell nucleus by 12 hours. But after 24 hours, the fluorescence in nucleus was dispersed while that in cytoplasm increased. Forty-eight hours later, the fluorescence weakened significantly in both cytoplasm and nucleus. Above all, the results afforded significant data for the study on function of DPV dUTPase gene and the pathogeny of DPV.

Study on Pathogenicity and Immunogenicity of ORF3 Gene Deleted Mutational Strain of Porcine Circovirus Type 2 to Piglets

YANG Xiao-nong;GUO Wan-zhu;XU Zhi-wen;ZHU Ling;WANG Xin;ZENG Xiu;WANG Xiao-yu

2008, 39(8): 1094-1099. doi:

Abstract ( 922 )

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Fifteen healthy piglets aged 28 to 30 day-old were divided into three groups at random(5 piglets per group), and were inoculated intramuscularly with mPCV2-C(ORF3 gene deleted mutational strain of PCV2), PCV2-CD(parent strain) and physiologic saline, respectively. By using the methods of PCR-RFLP and PCR, the DNA of mPCV2-C and PCV2-CD were found in the inguinal lymph nodes of slaughtered piglets from group A and B respectively, which proved that the mPCV2-C could replicate and proliferate in piglet body. Observation on histopathology showed that some pathological changes were found in the inguinal lymph nodes, liver cells and alveolar wall epithelium in slaughtered piglets of group A and B, in which that of group B was more serious, the results proved that the ORF3 gene deletion didn’t change the tissue tropism of PCV2. The dynamic determination of T lymphocyte subgroups in peripheral blood showed that, compared with group C, the percent of CD3⁺ and CD4⁺T lymphocyte subgroups of group A and B decreased, and the CD8+ cell of group A returned high above that values of group B and C, which indicated that ORF3 gene deletion weakened the influences of PCV2 on T lymphocyte subgroups, and mPCV2-C demonstrated the tendency to induce cellular immunologic response in piglets. The results mentioned above also indicated the decrease tendency of pathogenicity of the virus with ORF3 gene deletion. The dynamic determination of antibody against PCV2 proved that mPCV2-C induced the humoral immunity in piglets obviously, and had good immunogenicity. mPCV2-C is a candidate strain for gene deletion vaccine against PCV2.

A Histopathologic Study on Cysticercus pisiformis Infected Rabbits

SUN Xiao-lin;CHEN Huai-tao;CAI Xue-peng

2008, 39(8): 1100-1106. doi:

Abstract ( 2285 )

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In order to observe the histopathological changes of the rabbits infected with Cysticercus pisiformis, tissues of livers, hearts, kidneys and lungs from 33 naturally infected rabbits were collected. All the samples were fixed in the formalin and dehydrated with gradient alcohol. The 5-6 μm paraffin sections were made and stained with haematoxyylin and eosin, then were obser ved by light microscope. The results showed that zones of hemorrhaged necrosis caused by the migration of oncospheres of Taenia Pisiformis was formed on the surface of the rabbits livers, as well as a granulation tumor consisted of epithelioid cell and multiple nuclear giant cell. This special pathological change could be one of the important diagnosis points of the disease. Meanwhile, other pathological changes such as interstitial hepatitis, brightism, pneumonia and heart muscle fiber atrophy were also observed. This study layed the foundation for the elucidation of the characteristic pathological change of liver and the development mechanism of this disease.

The Distribution of Somatostatin Immunoreactive Cells in the Digestive System of Laiwu Black Goat

HOU Yan-meng;WANG Shu-ying;YIN Xun-he;HUANG Li-bo;FENG Hua-peng;LIU Guan-hua

2008, 39(8): 1107-1110. doi:

Abstract ( 1663 )

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The distribution of somatostatin (SS) immunoreactive(IR) cells in Laiwu Black goat digestive system were researched by immunohistochemical SABC method. The shape of SS-IR cells were mainly ellipse and cone . SS-IR cells showed peak density in the pylorus, but not detected in the cardia, colon, cecum and rectum. In pancreatic islets ,SS-IR cells were discerned predominately in the mantle regions,meanwhile SS-IR cells were also found in the exocrine portions and the central regions. In conclusion，SS-IR cells were showed up in the pylorus and the mantle regions of the pancreatic islets mostly.

Effects of Three Feed Additives on Mucosal Structure in Small Intestine of Broiler

WANG Ji-feng;LI Fu-yan;CHEN Yao-xing;WANG Zi-xu;GUO Yu-ming

2008, 39(8): 1111-1115. doi:

Abstract ( 1445 )

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Thirty-two healthy one-day-old AA broilers were divided into 4 groups in this study: Group A (control) with basic diet, Group B with basic diet and antibiotic (100 mg/kg of Colistin), Group C with basic diet and 500 mg/kg of Chinese Medicine (Youkangbao), and Group D with basic diet and 500 mg/kg of sodium butyrate.After seven weeks, the mucosal structure of small intestine was observed.Results were as follows: (1) There were significantly differences in the morphological change of mucosa of small intestine among A, B, C, and D groups.(2) The lengths of villi, V/C value, thickness of mucosa, and thickness of muscular layer in experimental groups were larger than that of control group. (3) The value of V/C decreased from duodenum to ileum in control group, but it was larger in ileum than that of the jejunum in experimental groups.However, the value of V/C was larger in experimental groups compared with control group.This study indicated that three feed additives could improve mucosal structure of small intestine in broiler; Sodium butyrate and Chinese Medicine (Youkangbao) have the feasibility to substitute of antibiotic in improving mucosal structure of small intestine.

Pharmacokinetics of the Main Component in the Compound Traditional Chinese Medicine Weeping Forsythia in Chicken

GAO Hai;ZENG Zhen-ling;LIU Kai-yong;JIANG Zhi-gang;XIANG Rong

2008, 39(8): 1116-1121. doi:

Abstract ( 1739 )

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Pharmacokinetics of Chlorogenic acid and Baicalin, two active components of the compound traditional chinese medicine Weeping Forsythia were studied in chickens after oral administration of a single dose (20 g/kg). The contents of chlorogenic acid and baicalin were 53 and 158 mg/kg in the compound. Drug concentrations in plasma were determined using high performance liquid chromatography (HPLC) assay. The pharmacokinetic parameters were calculated by 3p97 program. The pharmacokinetic process of Chlorogenic acid and Baicalin were all fitted with no-compartment model. The main pharmacokinetic parameters of chlorogenic acid and baicalin in plasma were as follows: Area under the concentrationtime curve (AUC), aera under the moment curve(AUMC),mean residence time (MRT),elimination half-life(t1/2),maximum drug concentration(Cmax) ,time of reaching maximum drug concentration(tmax) in plasma were （10.59±0.69） and （11.15±0.78）（mg/L）·h,(106.06±3.11) and (138.44±2.86)(mg/L)·h2,(10.01±0.68) and (12.42±0.43)h,(6.93±0.18) and (8.61±0.36)h, （0.98±0.25） and (1.04±0.25) mg/L，(1.93±0.15) and (2.45±0.19) h, respectively. The concentration change of active constituents components in plasma can reflect the metabolism rule of compound in chicken. The double peak of plasma concentration meant that there might be hepatoenteral circulation or stomach intestine circulation during the metabolism process. This study could provide references for the medicine in clinical application.

The Effect of Calcium Homeostasis Disequilibrium on the Impairment of the Neuron Mitochondria in the Chicken Brain Induced by Cadmium

LI Wen-jun;ZHAO Lan;XU Shi-wen

2008, 39(8): 1122-1126. doi:

Abstract ( 949 )

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To explore the mechanism of Cadmium Chloride on chondriosome damage of chicken neurons, neurons cultured in vitro were chose as the research object. Neurons were cultivated in the DMEM for 24 h with the final concentration of CdCl2 was 0,5,10,15 μmol/L,respectively. The inhibition ratio of neurons was evaluated with MTT colorimetric assay. The mitochondrial membrane potential (ΔΨm) was assessed by flow cytometry, and the calcium ion concentration within neurons(\[Ca²⁺\]i) was detected using the Fura-2/AM as the probe. The results showed that the cells inhibition ratio was increased, \[Ca²⁺\]i was increased, ΔΨm tended to degrade and the chondriosome cristae appeared ambiguity,fragmentation,disappearance, mitochondrial matrix was intumesced and vacuolized. The results indicated that CdCl₂ could cause the impairment of the mitochondrial membrane of neurons by increasing the \[Ca²⁺\]i , inducing the PT pore to open, degrading ΔΨm.

chODF Can Induce Osteoclast Formation from Chicken Embryo Marrow Cells

WANG Yan;HOU Jia-fa;HU Yun-feng;ZHANG Feng-rong;SHEN Xiang-zhen

2008, 39(8): 1127-1131. doi:

Abstract ( 877 )

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The purpose of this study was to investigate an effective culture method of osteoclasts(OC) formation and to test the possibility that chODF (chicken osteoclast differentiation factor) was sufficient for OC development in primary chicken embryo marrow cell culture. Femurs and humerus were dissected and bone marrow cells were collected and cultured in seven culture methods according to the medium with different concentration of chODF, M-CSF and chOPG. OC formation was observed and indentified by TRAP (tartrateresistant acid phosphatase), HE, toluidine blue staining, phasecontrast microscope and scanning electron microscopy. Results showed that the combination of chODF and M-CSF could induce OLCs formation and bone resorption in chicken embryo marrow cell culture. The number of TRAPpositive multi-nucleated cells in the culture medium increased dosedependently with ODF concentration (P<0.05), at meanwhile, OPG can interrupt the formation of OLC. In conclusion, we established an effective way to form OLC, and it could be employed to investigate the differentiation and activation of OC. It also provide theory of mechanism of OPG/ODF/RANK in the process of osteoporosis in layer hens.

The Study on the Expression Level of SYCP3 mRNA in Yak and Cattle-yak Testis

QU Xu-guang;LI Qi-fa;LIU Zhen-shan;DONG Li-yan;LI Xin-fu;HAO Chen-li;XIE Zhuang

2008, 39(8): 1132-1136. doi:

Abstract ( 1792 )

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The expression level of SYCP3 in yak and cattle-yak was studied by RT-PCR and real-time quantitative PCR in this paper. The results showed that SYCP3 was specifc expressed in yak testis, and the difference of expression level between yak and cattle-yak was extremely remarkable (P<0.01).The tissue sections of testis and epididymis showed that there wasn’t secondary spermatocytes or spermatids in cattleyak testis, and sperm could’t be observed in the epididymis. However, there were all levels of spermatogenic cells in yak testis, and sperms grew well in epididymis which was the same represent as man and mice lack or low express with SYCP3. SYCP3 gene was likely to have a relationship with male infertility of cattle-yak.

Analysis on Partial Sequence of Mitochondrial DNA D-loop Region Variation Position of Goat

WU Yan-ping;GUAN Wei-jun;ZHAO Qian-jun;HE Xiao-hong;PU Ya-bin;HUO Jun-hong;AO Hong;LI Kui;MA Yue-hui

2008, 39(8): 1137-1141. doi:

Abstract ( 899 )

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In this study, we aligned and analyzed 453 bp segment of mitochondrial DNA D-loop hypervariable region (Positions 15 735-16 187 bp of complete mitochondrial genome) from 1 182 sequences, neighbor-joining (NJ) tree and network showed that the sequences were divided into four distinct mtDNA lineages A-D,and included 1 001,134,24 and 13 sequences, respectively.After the alignment of the sequences,nucleotide position of A，B, C and D lineage specific were acquired, and included 5,6,18 and 7 variation positions, respectively, moreover, 2 lineages common variation positions were found.

Continuous in vitro Cultivation of Mycoplasma suis

LI Xiao-yun;JIA Xing-lin;SHI De-shi;XIAO Yun-cai;HU Si-shun;LIU Mei;YUAN Zong-hui;BI Ding-ren;

2008, 39(8): 1142-1146. doi:

Abstract ( 964 )

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Organisms obtained from symptomatic pig with clinical signs typical of porcine eperythrozoonosis, were Mycoplasma suis (M. suis), as was confirmed by microscopic examination of Giemsastained peripheral blood smears as well as by PCR. A fragment of 781 bp was amplified and its similarity to a known sequence of M. suis（accession numbers AJ504999）was 99.4％. Continuous culture of the pathogen was established from the infected blood sample. The culture medium was PPLO broth medium supplemented with newborn calf serum, yeast extract, glucose ,etc.. The optimal dilution of red blood cell (RBC), ratio of serum and pH during incubation were evaluated. Cultivation was initiated in an atmosphere containing 5% CO₂ at 37 ℃. The infected RBCs became ghosts with parasites after 72 h of cultivation. No host erythrocyte was supplied in the cultures except initiation. Cultures were maintained in medium ,with a percentage of parasitized erythrocytes（PPE） nearly 100% for more than 280 days. The cultured organisms adhering to ghost was identified to be M. suis on the basis of light and electron microscopic examinations, identical gene sequences, infect negative RBCs test, antibiotics sensitivity test.

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