Acta Veterinaria et Zootechnica Sinica

Polymorphism and Genetic Effect of FUT1 Gene M307 on Some Immune Parameters in Sutai Pigs

BAO Wen-bin;YE Lan;PAN Zhang-yuan;ZHU Jing;HUANG Xue-gen;HUA Jin-di;WU Sheng-long;

2010, 41(10): 1219-1224. doi:

Abstract ( 1058 )

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This study was conducted to analyze genetic effect of FUT1 gene M307 on some immune parameters in foundation population of Sutai ETEC F18 resistant breeding pigs and aimed to provide a theoretical foundation for further anti-disease breeding of Sutai pigs. The genetic variations at M307 of FUT1 gene in foundation population of Sutai ETEC F18 resistant breeding pigs (total 576 individuals) were investigated by PCR-RFLP method. The relationship between FUT1 gene and some immune parameters were analyzed in this study. The results showed that M307 of FUT1 gene digested by Hin6Ⅰcould be divided into three kinds of genotypes, AA, AG and GG. The genotype frequencies of AA, AG and GG at FUT1 locus were 0.235, 0.609 and 0.156, respectively. The allele A was dominant allele, its frequency was 0.540. This locus deviates from Hardy-Weinberg equilibrium (P<0.01) in above foundation population. The individuals with resistant genotype AA were significantly higher than those individuals with AG and GG genotypes（P<0.05）in HGB and WTBC, while there were no significant difference between AG and GG genotypes (P>0.05). The individuals with resistant genotype AA were significantly lower than those individuals with AG and GG genotypes（P<0.05）in heterophil/lymphocyte(H/L) ratio, while there were no significant difference between AG and GG genotypes (P>0.05). For SRBC antibody titer, there were no significant difference among AA, AG and GG genotypes (P>0.05). These results demonstrated that the resistant populations to ETEC F18 with genotype AA also showed fine characters of stress resistance，AA genotype had good genetic effects on disease resistance traits of PWD and ED, as well as normal disease resistance.

Deletion Mutation of the Bovine SREBP-1c Gene of Jiaxian Red Cattle and Its Affecting on Growth Traits

HUANG Yong-zhen;WANG Ju-qiang;MA Gui-bian;ZHANG En-ping;HUAI Yong-tao;MA Liang;LEI Chu-zhao;NIU Hui;XIAO Jie;CHEN Hong

2010, 41(10): 1225-1231. doi:

Abstract ( 692 )

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The present study focused on screening the genetic variation within bovine SREBP-1c gene intron 7 and analyzing its effect on growth traits in 441 individuals belonging to Jiaxian Red cattle. In this study, the 84 bp deletion mutation of SREBP-1c gene was detected by using the DNA sequencing and agarose electrophoresis methods. The PCR products of SREBP-1c gene exhibited two genotypes WW and WD, and two alleles were revealed: W and D. Genotype WW was the predominant genotype and W was the predominant allele in the studied population. The frequencies of allelic W and D in Jiaxian Red cattle population were 0.865 1 and 0.134 9. Genetic indices (He, Ne and PIC) in this cattle population were presented low genetic diversity. The χ²test analysis demonstrated that the breed was in agreement with HardyWeinberg equilibrium (P>0.05). The association of the 84 bp deletion mutation of SREBP-1c gene with growth traits of Jiaxian Red cattle breed was analyzed. The animals with genotype WD had significantly higher body weight and heart girth than those with genotype WW (P<0.01 or P<0.05) at 12, 18 and 24 months. The animals with genotype WD had significantly greater body length than those with genotype WW (P<0.05) at 18 and 24 months. The individuals with genotype WD had significantly higher average daily gain than those with genotype WW (P<0.01) at 18 months. These results indicated that the growth stages and the selected traits should be comprehensively considered in the breeding practice of Jiaxian Red cattle. The genotype WD may have positive effect on growth traits. Therefore, these results suggest that the deletion mutation of bovine SREBP-1c gene may be a useful marker for growth traits in future marker-assisted selection programs in Jiaxian Red cattle.

Identification and Prokaryotic Expression of Duck Adiponectin Gene

XUE Mao-yun;DONG Biao;ZHANG Ying;YU Jian-feng;MENG He;GONG Dao-qing;

2010, 41(10): 1232-1239. doi:

Abstract ( 977 )

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The purpose of this study was to clone and characterize the duck Adiponectin gene, and to reveal the tissue expression pattern of Adiponectin mRNA in duck, and to express recombinant duck Adiponectin in prokaryotic cells. The sequence of duck Adiponectin gene was obtained by reverse transcription PCR, 5 ′-rapid amplify cDNA end and 3′- rapid amplify cDNA end (5′-RACE and 3′-RACE) methods, and its expression in various duck tissues was detected by semi-quantity RT-PCR. Recombinant plasmid was constructed and transformed into E.coli. to express the recombinant duck Adiponectin in prokaryotic cells. The three Adiponectin cDNA fragments were amplified from total RNA using RT-PCR, 5′-RACE and 3′-RACE methods, respectively. After cloning and sequencing the three fragments, a 1 374 bp full length cDNA sequence of duck Adiponectin gene was acquired, which contained a full 738 bp nucleotides open reading frame, encoding 245 amino acids. The deduced Adiponectin amino acid sequence of duck contained 22 glycine-X-Y repeats at the collagen repeats as found in the chicken and mammalian Adiponectin. The duck Adiponectin CDS shared 94.9% and 86.0% homology with goose and chicken, and about 64.2%-67.0% homology with mouse, human and dog. Meanwhile, the putative amino acid sequence of duck Adiponectin shares 95.5% and 84.0% homology with goose and chicken, and about 65% homology with the above mammals. The semi-quantity RT-PCR showed that Adiponectin expressions were detectable in all tissues, and the expression level of Adiponectin gene was high in adipose, muscular stomach, small intestine, heart and skeletal muscle, medium in lung, gland stomach, ovary and spleen, low in liver, kidney and diencephalons. The recombinant plasmid of pET41a-duck-adp was constructed and transformed into E. coli BL21（DE3）. The 30 ku recombinant duck adiponectin was successfully expressed in E. coli. The Adiponectin gene is conserved in animal evolution, and Adiponectin mRNA expresses differently in various tissues. The recombined duck adiponectin could be expressed in prokaryotic system.

FSH Regulates Boar Sertoli Cell Proliferation via cAMP and Ca²⁺ Influx

WANG Xian-zhong;ZHOU Yu-lan;ZHAO Bo-chuan;ZHOU Yin-tao;CHEN Yong-jun;ZHANG Jiao-jiao;WANG Yi;ZHANG Jia-hua

2010, 41(10): 1240-1245. doi:

Abstract ( 656 )

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This experiment was conducted to study mechanisms of FSH regulating Sertoli cell proliferation. Sertoli cells of 2-3 week-old piglet were used as cell models, and cell viability, ELSIA and Western blot were used to explain the effect of FSH. The results were as follows: (1) From 0 to 50 ng·mL^-1, with the increase of the concentration of FSH, Sertoli cell number showed an upward trend(P<0.05); at 50 ng·mL^-1, effect of FSH on Sertoli cell proliferation was most significant; the concentration of intracellular cAMP also increased, following the increase of the concentration of FSH(P<0.05); (2) FSH begun to activate ERK1/2 cascade reaction 5 min after its affecting and the activating model had a periodicity; ERK1/2 inhibitor (PD98059 and U0126) could significantly inhibit the FSH-induced Sertoli cell proliferation and the expression of PCNA (proliferating cell nuclear antigen) (P<0.05); (3) Both of FSH and Forskolin could enhance cell proliferation and ERK1/2 activation (P<0.05), while Rp-cAMP and L-Ca²⁺ channel inhibitor (Verapamil) significantly both inhibited the FSHinduced cell proliferation and ERK1/2 activation(P<0.05); Rp-cAMP and Verapamil showed synergism effect (P<0.05). These results indicated that FSH could activate ERK1/2 via cAMP and Ca²⁺,which increased the expression of PCNA, and promoted Sertoli cell proliferation.

Optimization of Electrofusion and Artificial Activation Protocols of Porcine Handmade Somatic Cell Nuclear Transfer

REN Zi-li;ZHAO Yan-ling;YANG Xiao-gan;LU Yang-qing;LU Sheng-sheng;LU Ke-huan

2010, 41(10): 1246-1252. doi:

Abstract ( 1307 )

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The objective of this study was to explore the optimal conditions of electrofusion combined with artificial activation influencing the development of zona-free porcine cloned embryos, and to lay the foundation for Bamamini pig somatic cell nuclear transfer embryo by handmade cloning (HMC). Cloning embryos were fused by one-step or two-step electrofusion and activated in CHX+CB for 4, 5 and 6 h, then cultured in WOWs for 7 d to observe the fusion and embryo development. The results were as follows：(1) The fusion rate of oocytes subjected to one-step electrofusion with parameter 2 (180 V·mm^-1, 30 μs×1) was higher than that with parameter 1 (160 V·mm^-1，30 μs×1) (65.31% vs 58.33%，P<0.05). (2) There was no significant difference (75.88% vs 81.20%，P>0.05) in fusion rate between the groups of oocytes subjected to one-step electrofusion with parameter 3 (the first：180 V·mm^-1，10 μs×1; the second: 85 V·mm^-1，80 μs×1 with an interval of 60 minutes) and parameter 4 (the first：200 V·mm^-1，10 μs×1 ; the second: 85 V·mm^-1，80 μs×1 with an interval of 60 minutes). (3) There was no significant difference in cleavage and blastocyst rates of cloned embryos between the groups of oocytes subjected to electrofusion with the parameter 4 and parameter 2 ((79.98% vs 72.66%), (13.14% vs 9.36%)，P>0.05). (4) There was no significant difference in the rate of cleavage and blastocyst development from oocytes, which were subjected to electrofusion, activated by CB+CHX for 4, 5 and 6 h (P>0.05). The results revealed that the procedure with parameter 4 of electrofusion activated by CB+CHX for 5 h was relatively suitable.

The Effects of Heat Shock on the Development of Bovine Parthenogenetic Embryos in vitro

XU Wei;TANG Shuang;LV Zhi-yi;SU Feng;HUA Song;ZHANG Yong

2010, 41(10): 1253-1259. doi:

Abstract ( 611 )

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The present study was conducted to investigate the stress response of nuclear or cytoplasmic components to heat shock, and the detrimental effects of heat shock on expression of zygotic genes stage, cell apoptosis and in vitro development of bovine embryos. The parthenogenetic embryos were incubated at 38.5 or 41 ℃ for 7 h, respectively; then the nuclei of embryos cultured at 41 ℃ were transferred into cytoplasts enucleated at 38.5 ℃ to construct heat shock nucleinormal cytoplasm embryos (HN embryos) and 38.5 ℃ nuclei were transferred into 41 ℃ cytoplasts to construct heat shock cytoplasm-normal nuclei embryos (HC embryos). The development rate of nuclei-cytoplasm exchange embryos, expression of zygotic activated genes and apoptosis of embryo cells were detected. In vitro culture showed that blastocyst formation rate for HC embryos decreased significantly compared to parthenogenetic embryos in control and HN embryos.Incomplete activation of zygotic genes was observed in nuclear exchange embryos, especially in HC embryos. It suggested that heat shock might disrupt some cytoplasmic factors required for zygotic gene activation. Apoptosis induced by heat shock was observed in both HN and HC blastocysts and the incidence of apoptosis in HC blastocysts was significantly higher than that in HN embryos. These results suggested that: although heat shock injuries exist in both nuclear and cytoplasmic components, cytoplasmic component is more susceptible to heat shock than nuclear component.

Effect of Chinese Medicine Prescription on Beef Cattle in Summer: Ⅰ. Finishing Performance, Physiological Parameters, Serum Hormone Level and Enzymatic Activity

WANG Wen-juan;WANG Shui-ping;;ZUO Fu-yuan;ZHOU Pei;ZHAO Jian-jun;ZHANG Jia-hua

2010, 41(10): 1260-1267. doi:

Abstract ( 729 )

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The aim of the study was to investigate the effects of Chinese medicine prescription on the finishing performance, physiological parameters, serum hormone level and enzymatic activity of the beef cattle in the natural environment of heat stress. Eighteen healthy cattle with the similar age, conformation and weight (about 300 kg) which crossbred by Simmental bull and native cow had were randomly allotted into three groups. One group was the control group and the rest were the treating groups. There were six cattle in each group with six replicates and one cattle per replicate. The experiment was designed by single factor random arrangement. The cattle in the control group were fed the basal diet. And the cattle in the two treating groups were fed the basal diets with different Chinese medicine prescriptions supplied with 0.2 kg per day per cattle, such as prescriptionⅠand prescriptionⅡ, which composed of Herba Agastachis, Rhizoma Atractylodis, Cortex Phellodendri and Gypsum Fibrosunm with different proportions, as a kind of concentrate additive, respectively. The heat stress of the cattle could be caused by the moist heat environment in the Three-gorge Reservoir District in summer. The start-stop time of the experiment were from 1 July to 8 September in 2009. The duration of the preliminary experiment was 10 d and the duration of the formal experiment was 59 d. The results showed that the experimental cattle survived in the condition of the moderate heat stress and suffered the feeding environment of high humidity and high temperature the temperature humidity index in the experimental period was 84.01 averagely; every prescription could protect the experimental cattle from the hazard of heat stress by improving the finishing performance, decreasing breathing frequency and mean body temperature; every prescription could decrease the blood serum concentrations of adrenocorticotropic hormore, aldosterone, corticosteroid and glucagon, and increase that of growth hormone, insulin, thyroid stimulating hormone, triiodothyronine and tetraiodothyronine; every prescription could increase the blood serum activity of alkaline phosphatase and α-amylase, and decrease that of glutamate-pyruvate and glutamicoxaloacetic transaminase, lactate dehydrogenase and creatine kinase; in conclusion, the applied efficacy of the prescriptionⅠwas better than that of the prescriptionⅡwhen these prescriptions as a kind of feed additive prevented the heat stress.

Epidemiological Survey of Hepatitis E Virus in Industralized Swine Herds

ZHANG Hou-yong;CHEN Dong-sheng;ZHANG Yi;WU Yi-quan;HE Qi-gai;LIU Zheng-fei

2010, 41(10): 1268-1273. doi:

Abstract ( 1079 )

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The infection and epidemical characterization of hepatitis E virus (HEV) in industrialized swine herds were investigated. Eighty serum samples collected from 4 pig farms were detected for anti-gG antibody by ELISA. Then from 2007 to 2009, clinical liver specimen and anal swabs were collected in Hubei province, Henan province, and Anhui province for HEV RNA detection by nest RT-PCR (RT-nPCR), PCR products were sequenced, and neighbour-joining method was utilized for phylogenetic analysis. 56 out of 80 serum samples were anti-HEV IgG positive. 5 out of 8 pig farms were HEV RNA positive and the positive rate was 62.50%. HEV RNA positive rate was 8.4% (35/415) in anal swabs, while HEV RNA positive rate was 15.55% (21/135) in liver. The minimal detection age of piglet is 7 days old and the maximum detection date is 84 days old. Two typical sequences were submitted to GenBank, the accession number is GQ202266.1 for a liver origin isolate and FJ445406 for an anal swab isolate. Phylogenetic analysis shows that the homology among 38 positive strains is 92%-100%, all the isolated strains belong to the genotype 4. The isolated HEV from the rectal swab strain (FJ445406) is at the same branch with swine-origin strain DQ279091, but the strain from the liver sample (GQ202266.1) exists on the same sub-brunch with a human strain. Taken together, our study shows that HEV circulates in pigs herds in central China, but generally speaking, HEV genotypes in this area remained stable and belong to the same genotype. Our data contributes the knowledge of HEV infection in industrialized pig farm and control of HEV spread between human and animal, especially pigs.

Co-expression of Porcine INF-γ Gene and Its Molecular Adjuvant Effects on the PPV VP2-PCV2 ORF2 Eukaryotic Plasmid

CHEN Yan-ling;XU Zhi-wen;GUO Wan-zhu;ZHU Ling;XU Kai;TANG Yu-xiang

2010, 41(10): 1274-1280. doi:

Abstract ( 635 )

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Three genes encoding interferon-gamma (IFN-γ), major epitope (47-85AA) of ORF2 of PCV2 and VP2 of Porcine parvovirus were cloned and inserted into eukaryotic vector of pCI-neo, the recombinant expression plasmid of pCI-IFN-γ.ORF2.VP2 was constructed. And then the constructed plasmid was transfected into MDBK cells by lipofectamine. The expressed product was tested by Indirect Immunofluorescence and RT-PCR. The recombinant plasmid pCI-IFN-γ.ORF2.VP2 and pCI-ORF2.VP2, the control plasmid pCI-neo, PBS, the PPV inactivated vaccine and PCV2 subunit vaccine were injected into mice by intramuscular method. The lymphocyte transformation function of immuned mice was detected; In the different periods, the dynamic variation of blood T lymphocytes were assayed; PPV and PCV2 antibodies were measured too. The experimental results showed that the MDBK cells with the transfection restructuring plasmid could not only amplify the ORF2-VP2 and IFN-γ gene, but also produce green fluorescence which could be seen by Fluorescence Microscope. Expression of the specific protein was detected too. All of these were considered as a confirmation of biological activities of the expressed protein. From the seventh-day on, the splenic lymphocytes had obvious response to ConA in the group of mice immuned by pCI-IFN-γ.ORF2.VP2，which was higher than that of control and the group of mice immuned by pCI-ORF2.VP2; the number of CD3⁺CD4⁺, CD3⁺CD8⁺T lymphocytes were higher or significantly higher than that of control group and the group of mice immuned by pCI-ORF2.VP2; On the fourteenth day, the antibodies of PPV and PCV were detected. These results manifested that the pCI-IFN-γ.ORF2.VP2 effectively induced humoral and cellular immunity response in organism.

Enhanced Immunity to Killed FMDV Antigen by Dual Factors Recombinant Plasmid of Porcine Interferon-gamma and CpG Motifs

ZHAO Na;CHEN Guo-hua;FANG Yong-xiang;LI Wan-sheng;JIA Huai-jie;JING Zhi-zhong

2010, 41(10): 1281-1289. doi:

Abstract ( 597 )

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The present experiment was performed to determine whether a recombinant expression plasmid of IFN-γ and CpG ODN co-administrated with FMD antigen can enhance the immunological response. The mice were vaccinated intramuscularly with the conventional FMD antigen along with a recombinant plasmid inserted porcine IFN-γ gene and CpG ODN sequence. Specific antibody and neutralizing antibody of FMDV，lymphocyte proliferation，specific cytotoxic T lymphocyte (CTL) reaction and the secretory cytokines were detected. Adjuvant effect of the combination compared with CpG ODN alone on the antigen was able to elicit much stronger humoral immunity and cellular immunity in mice vaccinated，especially increase the neutralizing antibody level and CTL roles. Porcine IFN-γ and CpG ODN can enhance the immunological response against the FDMV antigen in mice，the recombinant plasmid of porcine IFN-γ and CpG ODN is potential to be an alternative adjuvant for the future FMD vaccine.

One Isolate of Natural Recombination between the S1 Gene and N Gene of Infectious Bronchitis Virus

XU Huai-ying;#;ZHANG Wei;#;QIN Zhuo-ming;QU Xin-ze;ANG You-ling;HUANGbing;LI Yu-feng;

2010, 41(10): 1290-1295. doi:

Abstract ( 989 )

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An infectious bronchitis virus (IBV) field strain named SC06 was isolated from a chicken flock with typical respiratory symptoms and nephritis lesions. The dwarf embryos were easily founded after inoculation with SC06 isolate in 10-day old SPF chicken embryos. 5 day-old SPF chickens were challenged with SC06 isolate, which could trigger respiratory symptoms and pathological changes, and the S1 and N genes of SC06 were cloned and sequenced. Phylogenetic analysis of the S1 and N gene sequences of the SC06 strain together with the prevailing domestic IBV isolates and the IBV references published in the GenBank revealed that SC06, 4/91 and 7/93B belonged to the same genotype Ⅱ in view of S1 gene, which disagreed with the prevailing IBV strains isolated by our laboratory in recent years, as well as domestic and other prevailing LX4 (genotype Ⅲ), which belonged to the genotype Ⅳ. However, SC06, DY06 and SDGE01 strains including other prevailing IBV isolates belonged to the same genotype Ⅲ in view of N gene, and different from 4/91 and 7/93B (genotypeⅡ). Alignment analysis of S1 nucleotide (amino acid) homology showed that the SC06 and the 4/91 from United Kingdom shared 98.7% (98%) homology, but only shared 75.6%76.0% with the most popular domestic IBV isolates; Further more, the SC06 shared 99.3% (99%), 93.4% (93.6%) homology with the domestic epidemic strain DY06, SDGE01 in view of the N gene nucleotide (amino acid) respectively, while only shared 85.7% (92.4%) with the 4/91. The results of this study suggested that the S1 gene of SC06 maybe originate from 4/91, and the N genes were related to the DY06 types of the prevailing domestic IBV strains, which indicated that the SC06 was a combination between the 4/91 strain and the prevailing domestic IBV strains such as DY06 isolate.

Establishment of the Diagnostic Method of IFAT for Cryptosporidium parvum Using the Recombinant Lactobacillus casei Expressing C. parvum P23

Geriletu;SHI Quan;WANG Yan-xia;LAN Li;MAN Da;ZHANG He-ping

2010, 41(10): 1296-1300. doi:

Abstract ( 664 )

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The present experiment was performed with the objective of establishing the diagnostic methods of IFAT and ELISA for detection of the Cryptosporidium parvum. The prokaryotic expression vector pGEX-P23 was constructed by DNA recombinant technology and expression of the GST-P23 fusion protein by IPTG induction. After purification, the fusion protein was used as coating antigen in the ELISA. Using the same method, the recombinant vector pMGP23 has been constructed and electrotransformed into the Lactobacillus casei, named as recombinant Lactobacillus casei, which was used as a known antigen in IFAT. 508 samples of bovine serum, 187 samples of sheep serum and 197 samples of goat serum collected from the animals in Inner Mongolia were detected by ELISA and IFAT. The positive rates were 0.59%(bovine), 5.88%(sheep) and 4.57%(goat) by ELISA and 0.19%, 5.88% and 4.57% by IFAT. Accordance rate of these two diagnostic methods in bovine serum samples was 33.33% and the rate in sheep and goat was 100%. The results showed that the recombinant P23 has a good reactionogenicity. The positive results of two diagnostic methods were basically identical, which showed good identity and sensitivity. The methods could be widely applied in the diagnosis of Ｃryptosporidiosis.

Studies on Extraction and Biochemical Characters of Surface Polymers on Traps of Nematodetrapping Fungus—Arthrobotrys oligospora

WANG Rui;YANG Lian-ru;YANG Xiao-ye;MA Ya-nan;HAN Hai-bing;ZHAO Xiao-hui

2010, 41(10): 1301-1305. doi:

Abstract ( 622 )

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This experiment was conducted to study the biochemical characters and effects of surface polymers from the nematode-trapping fungi, Arthrobotrys oligospora. The surface polymers were extracted by optimized method with 5 mol·L-1 LiCl, then proteins concentration, enzyme activity and toxicity of the surface polymers to nematodes in vitro were determined. Meanwhile, the effects of temperature, pH, chemical reagents and protease inhibitors on the surface polymers were also studied. The result showed that the protein contents and protease activity of the surface polymers from mycelium with traps was significantly higher than that from general nutrition mycelium and protease, acid phosphatase and alkaline phosphatase were detected for the first time. The optimal temperature was 65 ℃ for both polymers. The optimal pH were 7.0 and 6.5-7.5, respectively. They both had low toxity to nematodes tested. All above results indirectly confirmed the effects of surface polymers on the preying process, and a preliminary understanding of composition and biochemical characters of the surface polymers were carried out. It also provided valuable information for the studies on the mechanism of nematode-trapping fungi in future.

Effect of Monochromatic Light on Transcription of Opsin Gene in the Retinas and Pineal Glands of Broiler

JIN Er-hui;JIA Fei;WANG Zi-xu;CHEN Yao-xing

2010, 41(10): 1306-1311. doi:

Abstract ( 663 )

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Aim of this study was to compare the effect of monochromatic light on transcription of melanopsin(OPN4-1 and OPN4-2), green sensitive cone opsin (CHK-1) and blue sensitive cone opsin (CHK-2) genes in the retinas and pineal glands of broiler. Forty newly hatched male AA (Arbor Acres) broilers were exposed to white (400-760 nm), red (660 nm), green (560 nm) and blue light (480 nm) for 14 days by using LED (light-emitting diodes) with 15 lx light intensity as light sources, respectively. The total RNA of retinas and pineal glands was extracted respectively on the 14th day. After reverse transcription, PCR amplification was carried out to detect transcription of above four opsin-related genes by using corresponding primers. The results showed:(1)OPN4-1 mRNA expression of BL group were 13.44% and 16.77% higher than that of WL group either in the retinas or pineal glands (P<0.05) significantly, but OPN4-1 mRNA expression of RL and GL groups were 16.62% and 10.94% lower than that of WL group in the retinas (P<0.05) significantly and no significant difference was detected between GL and WL groups in the pineal glands (P>0.05).(2)In the retinas and pineal glands, OPN4-2 mRNA expression of RL group was the highest, followed by BL group, and GL group was the lowest in all light groups (P<0.05).(3)In the retinas, the expression of CHK-1 mRNA was higher than that of CHK-2 under four monochromatic lights. CHK-1 mRNA expression of GL group was 24.37% and 9.94% higher than that of RL and BL groups (P<0.05) and no significant difference was found between GL and WL groups (P>0.05). CHK-2 mRNA expression of BL group were 40.90% and 8.94% higher-than that of RL and GL groups (P<0.05) and no significant difference was found between BL and WL groups (P>0.05). Above results suggested that the different opsins have the different light color sensitivity. Compare to red and green light, blue light promote the transcription of OPN4-1 in the retinas and pineal glands.

Diversity of Total Activity of Nitric Oxidesynthase and Effect of Xuesaitongon the Diversity in Limbic System with Cerebral Ischemia Reperfusion Injury of Rabbits

ZHAO Jun;YIN Xun-he;LIANG Jing-yun;XIE You-lian;GUO Li-hong

2010, 41(10): 1312-1316. doi:

Abstract ( 615 )

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To research the diversity of total activity of nitric oxide synthase（NOS） and effect of Xuesaitong on that diversity in limbic system with cerebral ischemia reperfusion injury of rabbits, and to study the protection of Xuesaitong on cerebral ischemia reperfusion injury of rabbits. Sixty three 3-month-old Harbin White rabbits ((1 500±150)g) were divided in 3 groups: the cerebral ischemia treatment group, the cerebral ischemia untreatment group and control group, randormly. The analog of cerebral ischemia were established by operation and the divisity of total activity of NOS and the effect of Xuesaitong on it were determined by the biochemistry technique.The results indicated that the total activity of NOS increased at 2 h after cerebral ischemia treatment reperfusion, increased to the highest at 6 h, and then decreased from 24 h to 96 h, and recovered to normal level at 120 h last. The cerebral ischemia treatment group showed bigger extent to descend, and recovered to normal level at 96 h. The cerebral ischemia treatment group and control group is obviously lower than that in cerebral ischemia untreatment group(P<0.01). The results suggested that Xuesaitong could protect the brain tissue with cerebral ischemia reperfusion injury of rabbits by decreasing the activity of NOS and maintaining the content of NO.

Bioavailability of Marbofloxacin in Dogs by Oral Administrate

LIU Li-feng;GAO Li-li;SU Feng;JIANG Shan-xiang

2010, 41(10): 1317-1321. doi:

Abstract ( 996 )

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This experiment was conducted to investigate the pharmacokinetics and bioavailability of marbofloxacin in healthy dogs. A single oral or i.v. administration of the drug at the dose of 2.75 mg·kg^-1 to 8 dogs were performed in an open randomized crossover test. Blood samples were collected at different intervals after administration and concentrations of marbofloxacin were determined by HPLC method. The concentrationtime date were analyzed with 3P97 computer program to get the pharmacokinetics parameters.The drug concentration-time data were fitted to a no absorbed two-compartment open model after i.v. administration in pigs. The main pharmacokinetic parameters were as follows T1/2α(0.34±0.14)h，T1/2β (7.74±1.70)h，V(1.03±0.60)L·kg^-1， CL(0.22±0.08)L·h^-1； AUC(14.05±4.25)mg·mL^-1·h. The drug concentration-time data were fitted to a two-compartment model with first order absorption after single oral administration in dogs. The main pharmacokinectic parameters were as follows: T1/2α（(2.82±1.86)h， T1/2β（16.03±9.27）h， V/F(2.25±0.34)L·kg^-1，CL/F（0.21±0.05）L·h^-1，Tmax(1.55±0.62)h，Cmax（0.89±0.21）μg·mL^-1，AUC（13.66±2.77）mg·mL^-1·h. Marbofloxacin was rapidly absorbed and the drug was completely absorbed after single oral administration and have a good bioavailability in dogs.

Analysis on Apoptosis of Chicken Cecal Epithelial Cells Infected by E.tenella

GU Shao-peng;ZHENG Ming-xue;LI Bao-jun;HAN Ke-guang

2010, 41(10): 1322-1327. doi:

Abstract ( 630 )

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To study the damage mechanisms of coccidian infection with E. tenella, apoptosis of cecal epithelial cells from chicken with coccidiosis(E.tenella)was investigated. Chickens were infected with crude sporulated Eimeria oocysts. The pathological changes of cecal mucosa epithelial cells and enteraden epithelial cells were observed by histological section. Ultrastructural changes were monitored by transmission electron microscopy. Apoptosis was determined with TUNEL method. The results showed that at the 2nd day post infection, the number of apoptotic cecal mucosa epithelial cells was increased comparing with the control group. The most pathological changes occurred at 4-6 days post infection. Severe hemorrhage and exfoliated epithelial cells were found at the middle and posterior cecum. More seriously, cecal mucosa was entirely detached from some segment of the cecum with destroyed enteraden, degenerative or necrotic epithelial cells and erythrocytes. Other surviving cecal mucosa epithelial cells and enteraden epithelial cells showed apoptotic changes, such as cell shrinkage, chromatin condensation and aggregation at the periphery of the nucleons, nuclear fragmentation and forming apoptotic bodies. Apoptotic index showed extremely significant difference（P≤0.01） between the test and control groups. These results indicated that apoptosis of chicken′s cecal mucosa epithelial cells infected with E. tenella was more serious than the control group, specially at the epigamy phase. And the apoptosis severity is intimately related to the degree and course of coccidiosis.

Pathologic Diagnosis of Trichogenic Epithelioma on Hyline Commercial Layer Chickens

SUN Hong-lei;QIN Mei;XIAO Yi-hong;WANG Jian;YANG Feng;NI Wei;LIU Si-dang

2010, 41(10): 1328-1332. doi:

Abstract ( 932 )

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According to the characteristics of fatal disease of hyperplasia of skin and scabby among commercial layer chickens occurring in Shangdong province recently, epidemiological investigation, anatomic pathological change and observation, and histological and immunohistochemical examinations were carried out in suspected cases. The results showed that the disease was only discovered in the Hyline layer chickens of some batches of some chicken factory, about 1%-10% incidence，occurring among 5-day chickens, sustaining one month or so, finally leading to death. Yellow nodules could be seen initially throughout the skin, followed by the coalesced nodules, manking pachyderma and scab; histological examination indicated that the abnormal trichogenic tumor cells was invading the whole thickness of the dermis including nerve and striated muscle, and it was generally surrounded by fibrous tissue as well; Epidermal cells of skin lesions had a significant hyperplasia, and hyperplastic basal cell was atypia to some extent. Immunohistochemical examination revealed that in most of the tumor cells, CK was positive; Vimentin, CKLMW, CKHMW, P53, Ki-67 were negative; the hyperplastic connective tissue in the dermis were masculine for α-SMA. It could be concluded that the disease might be diagnosed as trichogenic epithelioma.

Research on Change of Acute Phase Protein and IL-6 in Cows with Endometritis

LI De-jun;Liu Yun-feng;PEI Xiao-ying;GUO Ding-zong

2010, 41(10): 1333-1336. doi:

Abstract ( 644 )

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To observe the changes of acute phase protein and IL-6 in the serum and uterine secretions in cattle with endometritis and search their significance on diagnose. Sixty-eight cows were selected and divided into three groups: N（Normal cows）, SE（Subclinical Endometritis）and CE (Clinical endometritis). The concentration of acute phase proteins and cytokine in serum and uterine secretions of endometritis cows were detected.The results showed that the concentrations of CRP, Hp, SAA and IL-6 in the serum and uterine secretions of cows with clinical endometritis and subclinical endometritis were higher than that of control group（P＜0.01）, and group CE was significant variable than group SE, but Hp and SAA were not detected in control group. The concentration of CRP, Hp, SAA and IL-6 were increased by different degree bacterial contamination in uterine and they can be the sensitive index for diagnosis with endometritis inflammation.

Identification and Sequence Analysis of MyoG Gene in Boer Goat (Capra hircus)

LIU Zheng-zhu;GONG Yuan-fang;ZHANG Chuan-sheng;FU Zhi-xin;ZHANG Wen-xiang;LI Fu-chun;FANG Xiu-min

2010, 41(10): 1337-1341. doi:

Abstract ( 688 )

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To explore the sequence structure and the regulation mechanism of goat MyoG gene in muscle differentiation, the partial DNA sequences (2 363 bp) were obtained for MyoG gene of Boer goat by direct sequencing, and the sequences were analyzed in other species. The results showed that the sequences of Boer goat MyoG gene consisted of 3 exons, 2 introns and some of 5′ (74 bp) and 3′ (260 bp) untranslated region, with 675 bp coding sequences, encoding a total of 224 amino acids. Structural analysis indicated that the peptide chain had not signal peptide, and a bHLH domain was located at No. 1-138 amino acids. Compared Boer goat MyoG with other species, the results showed that the homology of nucleotide of coding region and amino acid accorded with genetic relationship among the species. The cluster result of un-rooted phylogenetic tree was similar to the physiological characteristics of the species and their traditional classification. Identification and sequence analysis of Boer goat MyoG gene provided the important biological information for researching the mechanism of goat′s muscle development and regulation, artificially improving the muscle quality of goat.

microRNAs Expression in Adult Alpaca Skin

ZHU Zhi-wei;HE Jun-ping;CHENG Zhi-xue;WANG Hai-dong;LI Peng-fei;QIAO De-rui;DONG Chang-sheng

2010, 41(10): 1342-1345. doi:

Abstract ( 972 )

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The purpose of the present study is to investigate the effect of miRNAs abundance in adult alpaca skin and to discuss the role of miRNAs in skin and hair follicle. In the study, the level of miRNA abundance was investigated in alpaca adult skin by multi-species microarray screen. The result showed that expression of a total of 20 miRNAs was detected using the microarray screen in all samples. Based on the study, it was concluded that the miRNAs had important role in update, growth and development of skin and hair follicle.

The Comparative Analysis of Accuracy of Three Determining Endogenous Amino Acids Methods

ZHANG He-liang; LI De-fa;QIAO Shi-yan;ZHANG Zhao-qin;LI Ya-kui

2010, 41(10): 1346-1353. doi:

Abstract ( 628 )

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The aim of this study was to compare the accuracy of endogenous ileal amino acid loss values determined by regression (REG) method, the protein-free (PF) method and the homoarginine (HA) method. The endogenous ileal amino acid flows of growing swine determined using above methods were used to correct the apparent ileal amino acid digestibility (AID), and then judge the accuracy of endogenous ileal amino acid values by stability of collected digestibility values with the change of dietary protein concentration. Six barrows, with an initial body weight of 28 kg, were surgically fitted with simple T-cannulae at the distal ileum and fed six experimental diets containing crude protein 0, 5%, 10%, 15%, 20%and 25% according to a 6 × 6 Latin-square design. Each experimental period lasted 8 days. On day 6 of each period, ileal digesta wascontinuously collected for 24 h to determine AID and standardized true ileal amino acid digestibility (STID) using PF method. On day 8, the pigs were given a single meal of the HA diets and their ileal digesta was collected for 24 h in order to determine the HA flow and calculate real ileal digestibility (RID). The results showed that the values of endogenous ileal amino acid flow except glycine and proline determined using REG method were lower than those using PF method (P<0.1). Comparing with the HA method, PF method underestimated the loss of endogenous ileal amino acids (P<0.01) in the range of 10%- 25% of dietary protein concentration. The AID for all essential amino acids except phenylalanine and valine showed a quadratic increase (P<0.05) with an increase in dierary protein. And STID decreased (P<005) linely or quadraticly with an increase of dietary protein level. However, RID was stable and not influenced (P>0.05) by dietary protein level within 20% of CP. In conclusion, REG method and PF method underestimated the loss of endogenous ileal amino acids and HA method was more accurate than REG or PF method for determining endogenous ileal amino acid loss.

Isolation and Characterization of H3N2 Influenza Viruses from Ducks

ZHAO Guo;ZHONG Lei;ZHAO Kun-kun;LU Xin-lun;PENG Da-xin;LIU Xiu-fan

2010, 41(10): 1354-1358. doi:

Abstract ( 645 )

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From October 2008 to September 2009, 1 075 cloaca swab samples were collected from ducks in the Live-birds market, and 24 strains of H3 AIVs were isolated. The analysis of complete gene sequences and genetic evolution showed that two H3N2 avian influenza virus were both low pathogenic avian influenza virus and shared high genetic homology with the strains isolated from swine, whereas low to the strains from human. There are various degrees genetic recombination of the genes except HA, among different subtypes in different regions and different hosts, respectively.

Cloning and Sequence Analysis of the 16S rRNA Gene of Anaplasma bovis in Cattle

ZHOU Zuo-yong;;NIE Kui;TANG Cheng;HU Shi-jun;ZHOU Rong-qiong;ZHANG Ze

2010, 41(10): 1359-1363. doi:

Abstract ( 677 )

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In the study，we cloned and sequenced the 16S rRNA gene of Anaplasma bovis isolated from Zhongxian in Chongqing. The 16S rRNA gene of A. bovis was 1 412 bp in length. The sequence was submitted to GenBank (accession number: FJ169957). The 16S rRNA gene of A. bovis (FJ169957) had highest homology (99.0%) with that by Kawahara published (AB211163). In addition, 5 homologous sequences from A. marginale, 4 from A. centrale, 4 from A. bovis, 4 from A. ovis and 3 from A. phagocytophilum available were downloaded from GenBank. Phylogenetic analyses on Anaplasma 16S rRNA sequences were performed. The results indicated that A. bovis has closer genetic relationship to A. phagocytophilum than that to A. marginale, A. centrale and A. ovis.

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