Acta Veterinaria et Zootechnica Sinica

Research Advances on Electron Transfer Chain and Type Ⅱ NADH Dehydrogenases, NDH2s in Apicomplexa

LIAO Shenquan;CAI Jianping;QI Nanshan;WU Caiyan;LV Minna;YUAN Jianfeng;YU Jingshu;SUN Mingfei

2012, 43(1): 1-6. doi:

Abstract ( 420 )

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Infections with apicomplexan parasites are major causes of several parasitic diseases in human and animal. Unfortunately, the drug misuse has slowly led to the wide and serious drugs resistance in apicomplexan parasites, rendering serial drugs much less effective. Developing new and effective drugs against parasitic diseases has become extremely urgent. Many apicomplexans have an electron transport chain (ETC) superficially resembling that of the yeast Saccharomyces cerevisiae. The inherent difference between ETC pathways of the parasite and the host make it an appealing target for the development of novel drug. It has recently been demonstrated that an “alternative” or type Ⅱ NADH dehydrogenases (NDH2s) involved in ETC is completely lacking in host, NDH2s were proposed to be promising drug targets against apicomplexa. ETC pathways and NDH2s of apicomplexan parasites were reviewed.

Advances on the Studies of Parasite Proteasomes

CHEN Jia;;WU Songming;;ZHU Xingquan;HUANG Siyang

2012, 43(1): 7-13. doi:

Abstract ( 415 )

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Major advances have been achieved recently in the molecular composition of the proteasomes, their subunits, biochemical mechanisms and intracellular functions. Many cellular functions and processes are conserved in proteasomes including proteolytic functions, the regulation of cell cycle, apoptosis and transcription factor, enhancement of antigen presentation. In parasites, the proteasome is involved in the regulation of cell differentiation and replication, participation in impairment on hosts, and could therefore be an antigen inducing immune response in the host. Studies on parasite proteasomes will be beneficial for better understanding of mechanisms in host cell invasion by parasites, also providing a new thread for prevention and control of parasites. Therefore, this article reviews recent progress concerning the structure and functions of proteasomes, in particular, the parasite proteasomes.

Analysis of Gene Expression Profiles in Rugao Chicken Muscles during Different Periods

LUAN Deqin;CHANG Guobin;SHENG Zhongwei;HUANG Zhengyang;ZHOU Wei;GONG Linlin;CHEN Danyan;LIU Yan;WANG Kehua;DOU Taocun;CHEN Guohong

2012, 43(1): 14-21. doi:

Abstract ( 487 )

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The objective of this experiment was to study the development mechanism of muscle in Rugao chicken.The gene expression of Rugao chicken in muscle was analyzed during the different periods (212 week old) by GeneChip. The expression profiles of 13 379 differentially expressed genes were obtained, and the great changes of gene expression in muscle took place at chicken different growth stages. Compared with 2 week old, the number of downregulated genes at 4 and 12 week old were more than that of upregulated genes, while the number of downregulated genes at 6, 8 and 10 week old were less than that of upregulated genes. The clustering analysis of hybridization data was also carried out to indicate gene expression at different growth stages. The differential expressions of four genes were confirmed by realtime quantitative PCR, and the results indicated that the expression changes of these genes were generally consistent with the results of GeneChip. Expression profiles of different growth times were constructed in order to establish dynamic expression profiles, by which the responsive genes affecting muscle growtn were entirely acquired, and the growth and development mechanisms of muscle at different growth stages were also revealed in Rugao chicken.

Polymorphism of Gonadotropin Releasing Hormone Receptor (GnRHR) Gene and Its Relationship with Litter Size in Goats

HUANG Yanghe;WANG Pingqing;YANG Li;CHU Mingxing;ZHANG Baoyun;DENG Lamei;TAN Ying;FAN Qi

2012, 43(1): 22-28. doi:

Abstract ( 486 )

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The objectives of the present study were to search for genetic markers associated with high prolificacy by candidate gene approach and to give a scientific genetic basis for markerassisted selection for high reproductivity in goats. According to the Bos taurus GnRHR gene sequence, four pairs of primers were designed to detect single nucleotide polymorphisms of GnRHR gene in Boer and other nine breeds of goats in southwest China by PCRSSCP. The relationship between the polymorphism of GnRHR gene and litter size was observed in Chuandong White goat, Gulin Ma goat and Guizhou White goat. Only the sequences amplified by primers P1 showed polymorphisms. Genotypes (AA, AG and GG) were detected in different goat breeds. The sequencing results revealed that the genotype GG had one mutation (154G→A) compared to AA genotype. The goats with genotype AA have significant higher litter size than those with genotype GG and AG in three breeds of goats (P<0.05). And the goats with genotype GA were higher than those with genotype GG (P<0.05) in Gulin Ma goat. The finding showed that the GnRHR gene was related to the prolificacy of goats, which may be a factor affecting the reproduction in goats.

Sequence Analysis and Structure Prediction of the Promoter Region of MyoG Gene in Goats

LIU Zhengzhu;GONG Yuanfang;ZHANG Wenxiang;FU Zhixin;ZHANG Chuansheng;SONG Yu;MA Yanhua

2012, 43(1): 29-36. doi:

Abstract ( 489 )

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In order to explore the mechanism of goat MyoG gene transcripition regulation by studing the promoter region of the gene, the 5′flanking sequence of MyoG from the Bore goat genome was attained by PCR amplification, sequencing and sequence alignment. The DNA sequence of 969 bp was amplified. The content of G and C in the sequence was about 50.6%. The sequence analysis showed that there were one TATAbox at -24 bp and one GCbox, two CAATbox, two Ebox and one A+Trich motif. Meanwhile, some transcription factor binding sites including HSF, ADR1, cap were detected. The 5′flanking sequence of goat MyoG was compared with cattle, wapiti, human, mouse and chicken, the similarity was between 37.22% and 96.85%. It indicates that the 5′ flanking sequence of MyoG is comparatively conservative among different species. This work would lay the theoretical foundation for further study for expressing and regulating mechanism of MyoG gene in goat.

Cloning and Preliminary Functional Analysis of Bovine MyoG Promoter

WANG Qiuhua;CAO Yunkao;LI Shufeng;TONG Huili;XING Xiaoyou;LI Guangpeng;YAN Yunqin

2012, 43(1): 37-43. doi:

Abstract ( 438 )

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This experiment was conducted to compare the activity of different fragments size of Wagyu MyoG promoter and investigate the potential mechanism. According to the sequence of Bovine myogenin published in GenBank, the primers were designed to amplify the promoter region of Wagyu MyoG by performing PCR. The PCR products were ligated to pMD18TMyoGpro cloning vector. Positive clones were identified by restriction endonuclease and sequencing. Then the Wagyu MyoG promoter dualluciferase reporter gene vector(pGL3MyoGpro) was constructed, and then was transfected into muscle derived stem cells(MDSCs) and fetal fibroblast cells of Bos traurus, and the activity of dualluciferase reporter gene was detected. The activity and the specificity of these promoters during 1662 125 bp were identified in MDSCs. Bioinformatics analysis showed that the promoter fragment contained one TATAbox, fifteen Ebox and may contained binding sites of transcription factors MyoD, MEF2, MEF3, MTBF, TEF1, PRDF, Sp1, SRF, ERE, GRE and Oct1. By comparing the activity of different promoter fragments and analyzing on these transcription factors, it initially indicate that these transcription factors may play an important role in regulation of promoter activity. Cloning and analyzing of the promoter region and function of MyoG lay a foundation for further research on the mechanism of regulation and expression of MyoG in Bos taurus.

Parentage Inference with Single Nucleotide Polymorphism Markers in the Chinese Holstein in Beijing

GUO Gang;ZHOU Lei;LIU Lin;LI Dong;ZHANG Shengli;LIU Jianfeng;DING Xiangdong;ZHANG Yi;WANG Yachun;ZHANG Qin;ZHANG Yuan

2012, 43(1): 44-49. doi:

Abstract ( 447 )

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The aim of this study were to use single nucleotide polymorphisms (SNPs) markers for parentage inference in the Chinese Holstein, to discuss the effects of herd, birth year and birth month of the cow, sire family on the pedigree error rate, and to guide the cattle breeding and management. 255 highly polymorphic SNP markers, with the average minor allele frequency (MAF) over 0.45, were used for parentage inference among 84 Chinese Holstein bulls and 1 927 cows in Beijing. The analysis was carried out by the Cervus3.0 software adopting likelihood method. The average pedigree error rate was 20.9% in the experimental population. Moreover, the pedigree error rate among different herds, birth year and birth month of the cow were significantly different (P<0.05), while there was no significant difference among sires(P>0.05). Therefore, the occurrence of pedigree error was mainly due to the poor management in the dairy herds. It is very necessary to establish pedigree supervision and correction system and take measures to improve the pedigree accuracy, which would speed up the process of dairy genetic improvement in China.

Effects of Iloprost on in vitro Development of Preimplantation Porcine Embryos

WANG Feng;LIU Zhongfeng;SHI Chunrong;ZHANG Shuo;LV Ming;KONG Qingran;LIU Biao;LIU Zhonghua

2012, 43(1): 50-56. doi:

Abstract ( 391 )

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The aim of this study was to discuss the effect of PGI2 analogue iloprost on in vitro development of porcine embryos. In this study, at different times（0, 24, 48, 72 h）, iloprost with different concentrations（0, 0.2, 0.5, 1.0, 2.0, 5.0, 10.0 μmol·L-1）was added into the porcine zygote medium 3(PZM3) to observe the effect of iloprost on porcine IVF embryos and to screen the best additive dosage for embryo culture, blastocyst number and blastocyst cell number were counted at 156 h after fertilization. The speed of embryo fat metabolism and related genes(cox2, creb，pparδ, pdk, cpt2) expression level were tested for understanding the mechanism of iloprost promoting embryonic development. The results showed that addition of 2.0 μmol·L-1 iloprost at 48 hours after fertilization could significantly (P<0.05) improve the blastocyst rate (28%vs16%, P<0.05) and blastocyst cell number (49.42 vs 28.22, P<0.05), significantly accelerate fatty acid degradation speed (P<0.05) and promote the expression of lipid metabolism related genes(cox2, creb，pparδ, cpt2). However there’s no significant change in glucose metabolism related genes (pdk) expression level. These results demonstrates that iloprost can enhance the in vitro developmental capacity of porcine embryo by promoting fatty acid degradation.

Effect of Inhibin DNA Vaccine Harboring Two Copies of Inhibin Gene on Follicle Development, Production and Reproductive Hormones in Rats

WANG Shuilian;XUE Liqun;XING Zhaofang;CHEN Xiaojun;YANG Liguo

2012, 43(1): 57-64. doi:

Abstract ( 382 )

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Ninety rats were randomly assigned to five dosage groups of 18 each (T1, T2, T3, V, S, which was immunized with 10, 50, 100 μg·100 μL-1of pCISI, 100 μg pcMVS and 100 μL physiological saline per rat, respectively) to investigate the effect of double copies inhibin pcISI gene immunization on follicle development, production and reproductive hormones without any immune adjuvant. The results showed that the booster immunization could improve P/N value of antibody against inhibin, the average P/N value of antibody against inhibin of different dosages groups was higher than 2 after booster immunization, and the antibody level against inhibin in T3 group was higher than that in T1 and T2 (P <0.05) groups. Number of the mature follicles in the T3 group was higher than that in the control groups (P<0.05). The rats with positive antibodies had more 12.45 in number of mature follicles than that of rats with negative antibodies (P<0.05). In contrast to the control groups, litter sizes and number of the placentas were increased significantly in the pcISI immunization groups except for T1 group (P<0.05). 5.09 more placentas and 5.39 more litter sizes on average were also found in the positiveantibody rats than those in the negativeantibody rats (P<0.05). The levels of follicle stimulating hormone (FSH) and estradiol (E2) in the plasma of rats immunized with different dosages of pcISI plasmids were higher than that of the control animals, FSH and E2 levels in T3 group was higher than that in control group(P<0.05, P<0.01), there was no significant difference in P4 levels between different dosages groups and control group was observated(P>0.05). These findings demonstrate that double copies inhibin pcISI gene can enhance the immune response, stimulate FSH secretion and follicle development, improve the litter size without any immune adjuvant. The immune dosage of 100 μg·100 μL-1 is the optimum dosage in the study.

Effect of Dietary Crude Protein and Threonine on Feather Development of Starter Peking Ducks

GUO Feng;ZHANG Li;HOU Shuisheng;XIE Ming;HUANG Wei;YU Junying

2012, 43(1): 65-70. doi:

Abstract ( 473 )

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A 2×5 factorial experiment containing two CP levels(14%, 20%)and five threonine levels(0.5%, 0.58%, 0.66%, 0.74% and 0.82%) was conducted to evaluate the effects of different CP and threonine levels on feather development of starter Peking ducks. Four hundred and eighty onedayold male Peking ducks were randomly divided into 10 groups with 6 replicates in each group and 8 ducks in each replicate. These ducks were raised with experimental diets from 1 to 21 days of age. The results showed that threonine and CP in diets significantly affected the feather meat, one thousand downs weight, down length, one hundred plumages weight and plumages length(P<0.01). Dietary threonine levels from 0.74% to 0.82% did not affect feather reproduction rate, one thousand downs weight, down length, one hundred plumages weight and plumages length at higher dietary CP levels(P>0.05). However, dietary threonine levels from 0.74% to 0.82% significantly increased feather reproduction rate, one thousand downs eight, down length, one hundred plumages weight and plumages length at lower dietary CP levels(P<0.01). These results indicate that dietary threonine levels significantly affected feather development of starter White Peking ducks. Feather development need higher levels of threonine.

Effects of Tannin Content in Residue of Tealeaves on Digestion and Utilization of Nutrients and Metabolic Parameters of Nitrogen in Sheep

PAN Faming;LI Fadi;HAO Zhengli;ZHENG Chen;DONG Shuhui

2012, 43(1): 71-81. doi:

Abstract ( 441 )

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The present experiment was conducted to determine the effects of tannin content in residue of tealeaves(RTL) on digestion and utilization of nutrients and metabolic parameters of nitrogen in sheep. The contents of nutrients and tannin in RTL were analysed; The in vitro degradability（rdDM、tdDM、rdCP、tdCP） of dry matter(DM) and crude protein(CP) of mixtures with six ratio of RTL to soybean meal（0:10, 1:9, 2:8, 3:7, 4:6, 10:0 ）, which levels of RTL condensed tannin( RTLCT) of the mixtures were 0.00, 0.16, 0.31, 0.47, 0.63, 1.57 g·kg- 1DM, respectively, was measured by incubating in rumen fluid and rumen fluid + HClpepsin. To study the effects of RTLCT contents on digestion and utilization of nutrients and metabolic profiles of nitrogen in sheep, four High Mountain Finewool wethers (one and a half years old and average live weight of (29±2.55)kg) fitted with permanent rumen fistula and a 4×4 Latin square design with four cycles of diet each of 9 d for preliminary followed 6 d for sampling were used in this experiment. The contents of RTLCT in diets were 0.00(A), 0.77(B), 1.53(C), 2.30(D) g·kg -1DM . The results showed that: (1) the range of contents of CP, crude fiber(CF), TAE(tannic acid equivalent) and CT in three samples of TRL were 23.23%28.11%, 16.68%17.84%, 1.98%2.10%, 1.47%1.57%, respectively. (2) The degradabilities of DM and CP of the mixtures in rumen fluid and rumen fluid + HClpepsin(rdDM, tdDM, rdCP, tdCP) decreased linearly as the contents of RTLCT , the range of change were 61.39%25.67%, 79.61%26.47%, 60.76%16.68%, 84.85%17.69%, respectively. (3) In vivo, the intake of diet was affected by highraito of RTL, compared with diet A and B, the intakes of DM and organic matter (OM) of diet C (P>0.05) and diet D (P<0.01) were lower; the digestibilities of DM, OM, calcium (Ca), phosphorus (P), neutral detergent fiber (NDF) and acid detergent fiber (ADF) of diets were unaffected by the contents of RTLCT(P>0.05). But the excretions of urinary nitrogen were decreased linearly and retention of N increased as the RTLCT level, the retention of nitrogen of diet B, C, D were raised by 12.57% （P>0.05）, 23.61%（P<0.05） and 15.88%（P>0.05）than for diet A, respectively; there were no significant changes of the levels of pH and total nitrogen of rumen fluid (P>0.05) as the levels increasing of RTLCT, while the contents of NH3N and plasma urea nitrogen(BUN) linearly decreased, the levels of NH3N and BUN of diet C and D were 50.36 %(P>0.05) , 46.55%（P<0.05）and 68.04%, 59.99%（P<0.05）lower than that of diet A, correspondingly. It follows that residue of tealeaves were a matter with highprotein, lowfiber and middle content of tannin; under the condition of this experiment, the in vitro rdCP were decreased by RTL CT, but the digestibilities of CP in total gastrointestinal tract were unaffected by levels of RTLCT, and N retentions were increased owing to reducing of the excretion of N in urine as RTLCT; yet high ratio of residue of tealeaves in diet resulted in lowing of intake by sheep. Obviously, the protection effect of RTLCT with 1.53 g·kg- 1DM on protein was better, but no significant effect on the intake of diet.

Characteristics of Pathogenic and Antigenic Variation of H9N2 Avian Influenza Viruses HA S145N Mutants in China

CHEN Lu;ZHENG Luping;ZHAO Jun;WANG Chuanqing;WANG Zelin

2012, 43(1): 82-89. doi:

Abstract ( 466 )

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To investigate the effect of HA S145N mutation on pathogenicity and antigenicity of H9N2 avian influenza viruses (AIV), the EID50, ELD50, MDT, ICPI, IVPI and the duration of shedding virus from infected 8weekold SPF chickens of different H9N2 AIV strains with HA S145N mutation isolated from Mainland China were determined. The HI and VN activity of monoclonal antibody 2A4 and F6 on different H9N2 AIV isolates were assayed. The determined pathogenicity suggested that the H9N2 AIVs with HA S145N mutation have higher pathogenicity and can cause the death of SPF chickens. The 8weekold SPF chickens infected by H9N2 AIVs with HA S145N mutation shed virus earlier and last for a longer time. Monoclonal antibody 2A4 and F6 can inhibit the hemagglutinin activity of H9N2 AIVs with HA S145N mutation, however it can not neutralize the virus infection on CEF cells. The results showed that the H9N2 subtype AIVs have the tendency to evolve, resulting in the occurrence of mutants which have higher virulence and variable antigenicity. The site (S145) is one of HA protein epitope of the H9N2 subtype AIV. The higher virulent mutants may cause death of chickens and lead to more economic loss. The antigenic mutants may escape from immune clearance, which pose a new challenge to the immunoprophylaxis for the H9N2 avian influenza.

Comparison and Analysis of the Complete Nucleotide Sequence of FootandMouth Disease Viruses OT Strain

LV Zhanlu;WANG Guangxiang;SHANG Youjun;LIU Xiangtao

2012, 43(1): 90-97. doi:

Abstract ( 441 )

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Several pairs of primers were designed to amplify the complete genome of footandmouth disease virus (FMDV) OT strain by RTPCR and the complete genome was sequenced. Then all regions of OT strain genome were compared with those of the reference strains which sequences were obtained from GenBank, and their genetic characteristic were analyzed by molecular biological software. As a result, the genome of FMDV OT strain was 8 142 nt without poly(C) and, and was composed of 5′ untranslated region (UTR), openreading frame (ORF) and 3′ UTR, their nucleotide is 1 004 nt, 6 969 nt and 93 nt respectively. The pseudoknots area of OT strains loss 85 nt at 415499 region, but its 3A gene were not loss. And the OT strain shared closest homology with O/Akesu58 and OMⅢ, but OT strain derived earlier than O/Akesu58 and OMⅢ strains for many years. The origin and genetic derivation relationship of OT strain still need further attention.

Genetic Variation Analysis of the Canine Distemper Virus Based on Its H Protein Gene

GUO Ling;LEI Yan;CHEN Shijie;WANG Chengdong;YANG Xiaolong;YAN Qigui;

2012, 43(1): 98-104. doi:

Abstract ( 459 )

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Canine distemper disease caused by canine distemper virus (CDV) infection of domestic dogs and many carnivores is a deadly infectious disease. The article discussed the genetic characteristics, evolutional situations and epidemic regulation of CDV from molecular level. In this paper, we selected H protein gene of CDV as the target gene to evaluate molecular genetics, variation and epidemic regulation by compared the sequence of wild isolates to vaccine strains，the wild isolates included 14 CDV isolates from different geographical areas of China during the years 2002 to 2010, 12 CDV isolates around the world in 2006 to 2007, 12 CDV isolates from different hosts and the attenuated vaccine strains consisted of vCh, vCon, vOnder and vLed. The results showed that the nucleotide similarities of H gene between wild isolates and vaccine strains were from 86.2% to 92.1%, and the similarities of the deduced amino acid of H protein were 89.1% to 91.9%; Only AsiaⅠhad the Nglycosylation sites of H gene at 584 site; There were a higher nonsynonymous substitution rates in 3555 encoded amino acid sequences. And it indicate that antigenic variation of H protein may result in reduced effectiveness of attenuated vaccine, which couldn′t provide complete immune response preservation for some CDV epidemic strains.

Expression of a Chimera of Bovine Prion Protein and Yeast Ure2p Prion Domain in Escherichia coli and Preparation of Monoclonal Antibodies Against Prion Protein

LI Yanxin;MA Guiping;YANG Hanchun;LIU Quanguo;SHI Xiju;LI Bingling

2012, 43(1): 105-111. doi:

Abstract ( 447 )

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The recombinant expression plasmids pETPrPUPD was constructed by inserting Ure2p prion domain (UPD) gene into the recombinant vector pETPrP, The target gene was successfully expressed in the host cell BL21 (DE3) when induced with IPTG. Specific reaction was found between expressed protein and monoclonal antibody 6H4 by Western blot assay. The purified PrPUPD fusion protein was polymerized into amyloid fibrils, and exhibited partial resistance to proteinase K, Which similar to the structural characteristics of prion. The BALB/c mice were immunized with purified recombinant PrPUPD fusion protein. Immunized splenocytes were collected and fused with SP2/0 myeloma cell line. Hybridoma culture supernatant was screened by ELISA. Three hybridoma cell lines designated 1G3, 3A4 and 4D1, secreting monoclonal antibodies (mAbs) against recombinant bovine prion protein, were obtained. 1G3 could secret mAbs against bovine cellar prion protein and its immunoglobulin subclasses were IgG2a. The ELISA titers of the ascites fluid were 1:10 000. The specificity of the mAb 1G3 was confirmed with Western blot.

Effect of Moniezia benedeni on Local Mucosal Lymphoid Tissues of Small Intestine in Infected Sheep

YANG Yicheng;WANG Wenhui;QI Shanshan;HE Wanhong;Zha xi ying pai;ZHANG Wangdong;ZHANG Linjiang

2012, 43(1): 112-118. doi:

Abstract ( 435 )

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To study the effect of Moniezia benedeni on local mucosal lymphoid tissues of small intestine in natural infected sheep, the intestinal tract of adult sheep (infected group) which natural infected by Moniezia benedeni were observed from macroscopic, microcosmic and submicrocosmic scales, and then compared with the normal ones (normal group). The results indicated that the average length of parasite in intestinal tract was 1.5 m, and the scolex mainly absorbed in the place where there was abundant Peyer’s patch of jejunum. Generally, the number of parasite was about 12. Opening the jejunum, it was easy to see that themucous membrane which lived by parasite increased thickness and was covered by many gray mucus, and there were also some punctuate hemorrhage on it. The results of microscope showed that the local mucosal epithelium was desquamate, and the number of epithelial cells, intraepithelial lymphocyte and goblet cells were all increased obviously under the integral mucosal epithelium; there were hyperemia in capillaries of lamina propria, and hyperplasia of lymphocyte, plasmocyte, diffuse lymphoid tissue and goblet cells of intestinal gland happened in different degree. There also had mortification in the part of intestinal gland in the place which located by scolex; under the mucous membrane, there was obvious hyperplasia in lymphatic nodules and Peyer’s patch, and part of this hyperplasia that protruded into lamina propria could form some new dome areas; There were a great quantity of eosinophilic granulocytes infiltrated among lamina propria, mucosal substratum and mucosal layers. Under scanning electron microscope, the mucosal epithelium of intestine was desquamate in infected group; The scolex which was covered with layer of compact microvillus of Moniezia benedeni was ellipsoid and had four sucking disk but without rostellum and canaliculus. The findings suggested that the thickening of intestinal mucous membrane was mainly caused by hyperplasia of autologous tissue against the uninterrupt stimulation from the microvillus at the surface of parasite. So it maybe the mechanism of to resist infection of Moniezia benedeni in adult sheep.

Analysis of Genetic Variation in Different Populations of Cysticercus tenuicollisBased on the Mitochondrial Cytochrome b Gene

HAO Guiying;YANG Guangyou;GU Xiaobin;YANG Aiguo;YANG Yingdong;WANG Shuxian

2012, 43(1): 119-125. doi:

Abstract ( 389 )

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The objective of this study was to examine genetic similarity of C. tenuicollis among populations from different regions, and the phylogenetic relationships with other taenia species. The complete sequences of mitochondrial Cytochrome b (Cyt b) gene were used to investigate the intraspecific genetic subdivision of 33 C. tenuicollis isolated from seven different cities in Sichuan and Yunnan Province, NJ tree and MP tree were constructed using the MEGA version 4.0 software and PAUP version 4.0, respectively. The results showed that the complete sequence of Cyt b gene of C. tenuicollis was 1 068 bp, and 22 haplotypes were detected, homology analysis indicated that the homology identity levels of nucleotides of Cyt b gene among haplotypes were high, ranged from 97.3% to 99.9%, but the homology identity levels of nucleotides of isolates in this study and other 5 kinds of taenia spp. cestodes in taeniidae were below 82%. The phylogenetic trees based on Cyt b gene showed that all the haplotypes of seven populations were formed a single cluster that could be divided into three subgroups. There was no direct relationship to geography or host, appeared disordered distribution pattern which there was not obvious geography cluster or host cluster. There were some genetic differences in Cyt b gene within C. tenuicollis, but no significant variation, while interspecies difference was obvious. It was concluded that Cyt b gene can be used as genetic marker for population genetic studies of cestodes. The results of the present study provided foundation for further studies of population genetics of C. tenuicollis, and for the differential diagnosis of taeniid tapeworm infections.

Effects of Recombinant Chicken Interferonγ on Lipopolysaccharide Induced Immunological Stress in Broilers

CHEN Peng;SHAN Anshan;SUN Jinhua;CHENG Baojing

2012, 43(1): 126-132. doi:

Abstract ( 499 )

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The objective of this experiment was to research the effects of injection recombinant chicken interferonγ (rChIFNγ) in the prophase on immunological stress broilers. Forty 14dayold healthy AA broilers were allotted to 4 groups randomly which were group A, group B, group C and group D. The broilers of group A and group B were injected with 1 mL PBS at 14, 21, 28 and 35 days of age. At the same time, the group C was injected with 50 μg rChIFNγ (in 1 mL PBS) and the group D was injected with 100 μg rChIFNγ (in 1 mL PBS). At the time of 36dayold, 38dayold and 40dayold, the broilers of group B, group C, group D were injected with PBS (1 mL) which contained LPS (250 μg·kg-1 BW). At the same time, the broilers of group A were injected with PBS (1 mL) only. The performance indices were recorded at the time of 36dayold. Recorded the weight and intake of each broilers from 36 to 41 days. Hematologic indices and Tlymphocytes proliferation of broilers were assayed at 12 hours post the third injection of LPS. The concentration of interleukin 1 (IL1) and corticosterone (CORT) in the serum were measured at 12, 24, 36 hours post the third injection of LPS. The results showed that: (1) during the challenge, the growth performance was not affected by rChIFNγ (P>0.05); (2) 12 hours after the third injection of LPS, the white blood cell counts of group C and group D were higher than that of group A (P<0.05) and group B (P<0.05); (3) 12 hours after the third injection of LPS, the Tlymphocyte stimulation index of group C and group D were higher than that of group A and group B (P<0.05); (4) 12 hours after the third injection of LPS, the CORT of group A was lower than that of group B (P<0.05); 24 hours after the third injection of LPS, the CORT of group A was lower than that of group B (P<0.01), group C (P<0.05) and group D (P<0.05), 36 hours after the third injection of LPS, the CORT of group A was lower than that of group B (P<0.05); (5) 12 hours after the third injection of LPS, the IL1 of group A was lower than that of group B (P<0.05), 24 hours after the third injection of LPS, the group A was lower than that of the other groups (P<0.01). As the results shown, injection of rChIFNγ at prophase could release the IL1 and CORT in the immunological stress broilers, improve the cell immunity during the stress.

Histological and Ultrastructural Observation on the Pharyngeal Tonsil and the Tubal Tonsil of Goat

LI Zhengping;LI Pengcheng;YANG Qian

2012, 43(1): 133-137. doi:

Abstract ( 447 )

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The healthy goats aged 10 months were used to study the histology and ultrastructure of the pharyngeal tonsil and the tubal tonsil of goat by using histological and electron microscopy techniques. The results showed that the stratified squamous epithelium covering the tonsils was lesser, even some areas were covered by simple epithelium, and the intercellular junction of the epithelium was looser. There were many microvilli on the surface of the epithelial cells. Underneath the epithelium and in lamina propria were frequently infiltrated heavily by lymphocytes. The lamina propria consisted of scattered lymph nodules, aggregations of lymphocytes and diffuse lymphoid tissue, while a large distribution of lymphatic and high endothelial venules (HEVs) could be found in the diffuse lymphoid tissue. Furthermore, some dendritic cells (DCs) could be observed underneath the epithelium or in the lymphoid follicles. Our results indicated that the pharyngeal tonsil and the tubal tonsil could be used as an effective induction site or effective site of intranasal immunity in goat.

Measurement and Purification of GSHPx in Different Physiological Period of Sika Deer Blood

ZHANG Guangchao;WANG Xiaosong;MA Zefang;TANG Chao;CUI Kai;LIU Yongju;JIANG Xiaoming

2012, 43(1): 138-144. doi:

Abstract ( 452 )

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The contents of GSHPx in the whole blood of sika deer within different physiological period and the technique for purification GSHPx, were studied for developing antiaging resources of deer blood and laid a theoretical basis. The contents of GSHPx in the blood cells, plasma, dissolved blood and cell membrane were detected by using DTNB method and the contents of GSHPx in the whole blood were calculated. A 10%100% of saturation (NH4)2SO4 saltingout and column chromatography technology were used to purify the GSHPx and the SDSPAGE was used to measure its subunit molecular weight. The results showed that the highest contents of GSHPx appeared during the time of antler developing period ((1 973.07±25.43) U·mL-1), which was significantly higher (P<0.01) than that in the service period ((1 727.74±12.46) U·mL-1). However, no significant difference (P>0.05) was found between the value in antler developing period and the preantler period ((1 961.83±16.54) U·mL-1). GSHPx initial specific activity was 24.9 U·mg-1 and 0.37 mg·mL-1 purified GSHPx was achieved. Finally specific activity was 1 347.7 U·mg-1 and 54.1 times purification rate and 25% recovery rate were achieved. The relative molecular weight of subunit for GSHPx was 17.2 kDa. The results demonstrated that there are some certainly correlation between the change of GSHPx content and the physiological period of the deer. The saturation of 40%80% (NH4)2SO4 is the best condition for GSHPx purification.

Effects of Pulsatillae Decoction and Its Main Ingredients on LPSinduced NO，Eselectin and IL8 Secretion of Endothelial Cells

HU Yiyi;MU Xiang;HU Yuanliang

2012, 43(1): 145-151. doi:

Abstract ( 427 )

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The aim of the research was to investigate the antiendotoxin of Pulsatillae Decoction (PD), the levels of NO，Eselectin and IL8 secreted by cultured rat intestinal microvascular endothelial cells were determined after treatment with its active ingredients, namely anemoside B4, anemonin, berberine, jatrorrhizine, palmatine, aesculin, and esculetin. Endothelial cell was challenged with 1 μg·mL-1 LPS for 3 h, then treated respectively with PD at three concentrations (1, 5 and 10 mg·mL-1) and its seven ingredients at three concentrations (1, 5 and 10 μg·mL-1). The cells were incubated at 37 ℃ in a cell incubator for 21 hours. The supernatants were collected and the content of NO，Eselectin and IL8 were analyzed by ELISA kits. The results showed that as compared with LPS control group, the NO contents were significantly decreased in the middle and highdose groups of PD, the middle and highdose groups of Anemonin, the middle and highdose groups of Berberine and the highdose group of Esculetin (P<0.01 or P<0.05); The Eselectin contents were significantly decreased in the highdose group of PD, the highdose group of Anemoside B4, the highdose group of Berberine and the middle and highdose groups of Jateorhizine (P<0.01 or P<0.05); The IL8 contents were significantly decreased in highdose group of PD, the middle and highdose groups of Aesculin and the middle and highdose groups of Esculetin (P<0.01 or P<0.05). It suggested that PD may exert beneficial therapeutic actions on systemic inflammatory response syndrome, multiple organ dysfunction syndrome and immune diseases mediated by NO, Eselectin and IL8.

Genetic Effects of IGFI and IGFBP1 Genes on the Growth Traits in Jinghai Yellow Chicken

ZHAO Xiuhua;WANG Jinyu;ZHANG Genxi;WEI Yue;GU Yuping;YU Yabo;SHI Huiqiang

2012, 43(1): 152-158. doi:

Abstract ( 478 )

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The aim of this study was to analyze the association of SNPs in IGFI and IGFBP1 genes with chicken growth traits.PCRSSCP approach was applied to assess the single nucleotide polymorphisms(SNPs) and the general linear model was used to analyze genetic effects between different genotypes and growth traits of the Jinghai Yellow chicken. For the IGFI gene, three genotypes (AA, AB and BB) were detected in Jinghai Yellow chicken population, the frequency of A and B alleles was 0.613 and 0.387, respectively. Sequencing revealed one mutation (G60A) of IGFI gene in the genotype BB in comparison to the genotype AA. For IGFBP1 gene, three genotypes (CC, CD and DD) were detected in Jinghai Yellow chicken population, the frequency of C and D alleles was 0.558 and 0.442, respectively. Sequencing revealed two mutations (A21T, T104C) of the IGFBP1 gene in the genotype DD in comparison to the genotype CC. The results showed that individuals with BB genotype of the IGFI gene had higher bodyweight at 4 weeks compared to individuals with AA and AB genotypes (P<0.05); The individuals with DD genotype of the IGFBP1 gene had significantly higher bodyweight at 4, 12, 16 weeks compared to the individuals with CC genotype (P<0.01). The results suggest that SNPs of IGFI and IGFBP1 genes have certain effects on growth traits of Jinghai Yellow chicken, which can be used in markerassisted selection to accelerate the chicken breeding progress.

Optimization of Electroporation for Transfection of Goat Mammary Epithelial Cells

DU Shan;MA Baohua;LI Liangliang;LIU Jun;ZHAO Xiaoe;ZHANG Yong

2012, 43(1): 159-166. doi:

Abstract ( 502 )

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This study was conducted to obtain the optimal electroporation parameters for transfection of goat mammary epithelial cells. Mammary epithelial cells of diary goat were cultivated successfully using tissue mass inoculation method, which were identified by immunofluorescence staining of cytokeratin 18 and detecting the growth curve. And then the effectiveness of different voltage (120, 140, 160, 180, 200 V), different pulse length (5, 10, 15, 20 ms), and two different cell lines (GMEC1, GMEC2) of the first generation, 3rd generation, 5th generation, 9th generation were compared. Finally, it was found that one 15 ms pulses of 180 V can introduced DNA into the cells effectively. The results inferred that passage numbers exert more remarkable influence on transfection efficiency than genetic background. The lowpassage mammary epithelial cells had the highest transfection efficiency. The ability of these electroporation parameters to produce stably transfected goat mammary epithelial cells was determined following G418 selection with transfecting another vector, the goat mammary epithelial cell clonies was got, which express the exogenous humanbetadefensin3 gene stably. This study provides the optimal electroporation parameters for introducing DNA into mammary epithelial cells and improves the efficiency of transgenic cloning by SCNT.

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