China Animal Husbandry and Veterinary Medicine

Effect of Supplementation of Bacillus subtilis Natto on the Development of Digestive Tract in Calves

ZHANG Hai-tao;WANG Jia-qi;BU Deng-pan;LUAN Shao-yu;DENG Lu-fang;ZHOU Ling-yun;ZHOU Zhen-feng;WEI Hong-yang;SUN Peng

2010, 37(1): 5-9.

Abstract ( 1796 )

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The experiment was conducted to study the effects of Bacillus subtilis Natto  on the development of digestive tract in pre-weaning and post-weaning calves. Twenty-four Holstein male calves were used in a complete randomized block design, and was assigned to one of three treatments, 8 calves in each group. The calves in the two experimental treatments were fed Na and N1 Bacillus subtilis Natto while no Bacillus subtilis Natto was fed to the calves in the control group. The calves were fed with milk supplemented with the Bacillus subtilis Natto, and starter diet was also fed to the calves until weaning. Four calves in each group were slaughtered randomly at weaning and others were raised until 8 weeks after weaning. All the calves were slaughtered at the end of the trial. The results showed that compared with the control group, the weight of ruminoreticulum in calves fed Na Bacillus subtilis Natto was significantly enhanced (P<0.05); but the ruminoreticulum pH-value in the calves from N1 group was decreased throughout the experiment, while that of abomasums was dramatically decreased during the beginning period of weaning(P<0.05). The length of the duodenum in the calves fed Na Bacillus subtilis Natto was shorter than the control(P<0.05), so was that of the ileum (P<0.05). In addition, Bacillus subtilis Natto decreased the pH value in the duodenum and the pre-jejunum. In this study, we found that Bacillus subtilis Natto helped the digestion of the nutrients and promote the health of the calves.

Effect of Dietary Supplementation Potassium Chloride on the Production Performance and Blood Indicators of Dairy Cows in the Condition of Heat Stress

KUI Hua; Alateng Suhe;Turigen Baiyila

2010, 37(1): 10-12.

Abstract ( 1611 )

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45 Holstein lactaing cows with similar lactaion period, calving number and milk yield were randomly divided into 3 groups(n=15), in order to research the effects of diet adding potassium chloride on production performance and blood indexes of lactating cows during July to September. The diets of three experimental groups were basal diet(comprising corn silage, green hay and concentrates) (control group), basal diet＋50 g potassium chloride (groupⅠ) and basal diet＋100 g potassium chloride (groupⅡ) respectively. The results showed that there were no significant differences between groups in content of free fatty acids(NEFA), glucose(GLU) and urea nitrogen(BUN) in serum (P>0.05). β-hydroxybutyrate (BHBA) content was significantly higher in control group than in group Ⅰand Ⅱ(P<0.05),whereas no significant difference was found between group Ⅰand Ⅱ. Compared with the control group, decreasing degree of daily milk yield was less 1.6 kg (P<0.05) and 0.7 kg (P<0.05) for group Ⅰand Ⅱrespectively. There were no significant difference between groups in milk composition (P>0.05).The results indicated that potassium chloride supplementation can relieve the decreasing of milk production in summer.

Nutritive Peculiarity of Bile Acid and its Application in Poultry Production

LIU Jing-sheng;YANG Yu-zhi;WANG Jun-rong;FU Gui-hua;LI Yan-wu

2010, 37(1): 13-16.

Abstract ( 1422 )

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Bile acid is considered as the most important and most specific bile components. Bile acid has the important influence on the lipid metabolism.Bile acid affects the lipid metabolism in the liver， the principal role of bile acids is emulsification, and thus facilitation of the absorption of lipids as well as lipid-soluble matters.The research progress of bile acid in the poultry nutrition is discussed in this article.

The Nutritional Value of Corn DDGS and its Effect on Pork Quality in Growing-finishing Pigs

LI Gen-lai;YAO Wen

2010, 37(1): 17-21.

Abstract ( 1824 )

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Corn distillers dried grains with soluble (corn DDGS) is a co-product produced by dry mill ethanol plant. It’s a valuable source of crude protein, fat, available phosphorus and vitamin for swine. Recent years, with the development of processing technology, the quantity and quality of corn DDGS increases and has increasingly been included in swine diets. However, the high fat content in corn DDGS, especially unsaturated fatty acid, has detrimental effect on pork quality, especially fat quality and fatty acid composition in growing-finishing pigs. The paper reviews the nutritional value of corn DDGS and effect of corn DDGS supplementation on pork quality.

The Effect of Different Proportions of Organic Zinc and Inorganic Zinc on Growth Performance of Weaning Pigs

ZHANG Chun;KUANG Sheng-yao;TANG Ling

2010, 37(1): 22-24.

Abstract ( 1705 )

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This study explored the effect of different proportions of zinc amino acid chelates(Zn-AAC) and zinc sulphate(ZnSO4) on the growth performance of weaning pigs. Forty five hybrid pigs of 28 days old were selected and randomly allotted to one of five dietary treatments. There were three replicates per treatment and three pigs in each replicate. All animals were fed with the same basic diet supplied 100 mg/kg of zinc but with different proportions of organic zinc (Zn-AAC) and inorganic zinc (ZnSO4). Pigs in the control group were complemented with 100% ZnSO4， and the amount of zinc in diets of group 2， 3， 4 and 5 were substituted with 40%，60%，80% and 100% Zn-AAC，respectively.The 28-day trial included two phases of 1 to 14 d and 15 to 28 d.Data obtained in this study showed: from day 1 to day 14， the ADG of piglets in group 2，3，4 and 5 was increased by 22.0% (P>0.05)， 38.5% (P<0.01)， 32.6% (P<0.05) and 26.2% (P<0.05)， respectively， and ADFI， by 24.6% (P<0.05)， 25.3% (P<0.05)， 23.5% (P<0.05) and 23.8% (P<0.05)， respectively， when compared with the control piglets. From 15 to 28 d， all animals fed with organic zinc displayed higher ADFI than the control group (P>0.05). From 1 to 28 d， piglets fed diets with 60% of organic zinc had statistical improvement in ADG(P<0.01)， with no significant change in FCR. In conclusion， the supplementation of organic zinc with normal dosage could increase the ADG， ADFI. The combination of organic and inorganic zinc is superior to either one. The optimal proportion of organic zinc to inorganic zinc is 60∶40.

Effects of Soybean Trypsin Inhibitors on Histological Structure of Pancreas in Rex Rabbits

ZHANG Jian-yun;YI Zhong-hua;MA Qiu-gang;GUAN Shu;LIU Lai-ting;JI Cheng

2010, 37(1): 25-27.

Abstract ( 1620 )

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30 healthy (42±4) day-old weaned rex rabbits with the average initial body weight of (711.4±108.8) g were randomly allocated to 3 groups with 5 males and 5 females each group. All rabbits were raised in separated cages. The diets for each group contained same level of crude protein and energy. The diets contained 25% raw soybean, 25% chemically-processed soybean and 25% soybean-meal. The TI concentration (mensurated value) was 6.55, 1.99 and 0.81 mg/g respectively. The experiment lasted for 6 weeks, and at the end of the experiment 3 male rabbits randomly chosen from each group were slaughtered((91±4) day old, average body weight of 1544.2 g±114.8 g))after 4 to 6 hours of fast to take pancreas samples. These samples were fixed in 10% neutral formaldehyde solution, using conventional paraffin slice and HE dye for optical microscope observation. Samples for electronic microscope observation were fixed in 2.5% glutaraldehyde, conventionally resin embedded, very thin sliced, and uranium-plumbum dyed. Optical microscope observation showed the hyperplasia of fibroid connective very thin sliced, and uranium-plumbum dyed. Optical microscope observation showed the hyperplasia of fibroid connective tissue among the pancreas, oedema between lobes, disappearance of part of the gland, cell oedema, and border illegibility of the rabbits fed with 25% raw soybean. Abnormity of pancreas karyon, chondriosome oedema, disappearance of crista, partial membrane rupture, decrease of endoplasmic reticulum, and significantly cutting down or vanishing of particle secretion were discovered by electronical microscope observation in rabbits fed by 25%raw soybean diet. 25% raw soybean did not significantly increase the weight of pancreas, but abnormal changes occurred in histological structures. As for the other 2 groups, the results remained the same that no abnormal changes were found in histological structures. 

Purification and Partial Characterisation of Feruloyl Esterase from Barley Malt

LIU Sha-sha;LI Hua;SHI Qing-hua;WANG Xi-dong;CHENG Fa-xiang

2010, 37(1): 28-31.

Abstract ( 1315 )

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The activity of feruloyl esterase has been enhanced in barley grain and barley malt.Feruloyl esterase solution was extracted from barley malt has been purified by ammonium sulfate precipitation，DEAE-cellulose column chromatography.The feruloyl esterase was purified 34-fold as shown in SDS-PAGE gel electrophoresis and the molecular weight estimated to be about 29.3 kD.Characterization of the enzyme exhibited that the optimum pH and the optimum temperature were 5.5 and 50 ℃，respectively，and it lost little activity after one hour below 60 ℃ from pH 4.5 to 7.5.It was inhibited by Zn²⁺，Fe³⁺，Cu²⁺，and strongly activated by EDTA and Na⁺. 

Effects of Rating Polysaccharide by Alcohol in Astragalus Polysaccharides  on the Serum Interleukin-2 Contents in Chickens

LUO Yan;GU Xin-li;LU Zhi-zi;XU Lei;LI Zheng-guo

2010, 37(1): 32-33.

Abstract ( 1392 )

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To investigate the effect of rating polysaccharides by alcohol in astragalus polysaccharides on the serum interleukin-2 contents in chickens. By using three different concentration levels of alcohol to get rating polysaccharides in APS: A1, A2, A3; 320 one-day-old chickens were randomly divided into eight groups, experimental groups and control group were hypodermic injected A1, A2, A3, A1+A2, A1+A3, A2+A3, A1+A2+A3 and distilled water separately for seven consecutive days, and then draw blood on the 22nd, 29th, 36th, 43rd, 50th, 57thday to detect serum interleukin-2 contents, and evaluate the immune adjuvant effect of rating polysaccharides. A2, A3, A2+A3, A1+A2+A3 can increase serum interleukin-2 contents. The valid targets of APS are A2, A3.

Preparation of Specific Anti-FPV IgG from Horse

RAO Jia-hui;WANG Yu-ping;LEI Lian-cheng

2010, 37(1): 34-39.

Abstract ( 1775 )

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Feline panleukopenia is an infectious disease that infected more and more animals by FPV upward momentum. It has high pathogenicity and mortality. Treatment of the disease had not better and effective drug at home and abroad. The test selected horses as immune animals and extracted its serum. We optimized a new method of purification of IgG, by digesting the serum at 37 ℃ with pepsase, removing albumin in caprylic acid, and the twice ammonium sulfate precipitation. According to the character that Protein A and Fc of immunoglobulin can connect with IgG of most mammals, the F(ab′)2 was collected and purified by Protein A column filter.According to the relavant items of “People’s Republic of China States Pharmacopoeia”(2005 version). The physical properties, heat antigenicity, safety, aseptic, HA and HI test, agarose double diffusion, cells neutralization test and animal treatment test of this biological product of horse anti-FPV IgG were examined. The results showed that the product is a milk white solution that is non-layered, free of visible particles, bacteria, fungi, mycoplasma and other microbial contaminations. Rabbits, mice, and guinea pigs injected with the product were observed for 10 successive days. The results showed that the product is highly specific with the titer of 128-fold, good security and stability, non-toxic side effects. F(ab′)2 molecular weight of about 110 ku, agents accounted for 70.7% of total protein. The clinical symptoms of all the 5 cats disappeared 7 days later with the recovery rate of 100%, indicating the high effectiveness of the product in treatment of feline panleukopenia.

Cloning and Sequence Analysis of Genomic Termini of Duck Enteritis Virus

ZHOU Li-yuan;GUAN Fei;YU Ke-xiang;HUANG Bing;SHAN Hu;LI Yu-feng

2010, 37(1): 40-44.

Abstract ( 1355 )

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Two pairs of primers RTLT1 and RTLT2 were designed according to the published nucleotide sequence of the duck enteritis virus (DEV).The genomic DNA of DEV was extracted and self-lignased. 1607 bp and 1978 bp genomic fragments were amplified using PCR which were both cloned into pMD18-T vector. After identification of recombinants by PCR and Pst I digestion, the positive plasmids were sequenced. The result showed the sequences of the overlap region of two PCR products were identical. The G+C content of the overlap sequence reached 75%. Some direct and inverted repeated sequences and four characteristic sequences, α palindromes, β sequence surrounding the teminal joint, highly conserved γ sequences and AnTn sequence were found. Compared with the report of the DNA termini of equine herpesvirus 1, bovine herpesvirus 1, varicella zoster virus and pseudorabies virus, the sequence was confirmed to be the termini of duck enteritis virus genome. These sequence elements were proposed to be important for further understanding of DEV replication.

Cloning and Sequence Analysis of the Complete Genome of Porcine Circovirus Type 2 Gansu Lanzhou Strain

JIA Huai-jie;JING Zhi-zhong;FANG Yong-xiang;WANG Xiao-xia;CHEN Guo-hua;LI Ke-bin;DOU Yong-xi;QU Hui-ling;

2010, 37(1): 45-50.

Abstract ( 1519 )

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Two pairs of primers were designed and synthesized according to the full length genome of PCV2 in GenBank. Genomic DNA was extracted from samples of suspected post weaning multisystemic wasting syndrome (PMWS) pig in Lanzhou, Gansu province, and the full length genome of PCV2 was fragmentation cloned by PCR into pGEM-T easy vector, recombinant vector pGEM-PCV2 were constructed and then sequenced. Sequence analysis with DNAStar software showed that the genome of PCV2 Gansu Lanzhou strain is 1767 bp and the result of homology comparison of the full length genome among strains published in GenBank, indicated that it displayed high nucleotide and amino acid identities with ZhouKou (EU656143) strain (99.4% nucleotide identity and 98.6% amino acid identity), but distinct from GS03 (EU547458) which was another Lanzhou epidemic strain. So strain in this test may be a variant strain (GenBank accession No. FJ447482) prevalent in Lanzhou, Gansu.

Expression of N-terminal Major Epitope Domain of ORF2 of SHEV in E.coli

RAN Xu-hua;WEN Xiao-bo;WANG Mi;LI Dong-ye;HOU Xi-lin

2010, 37(1): 51-53.

Abstract ( 1379 )

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To express N-terminal major epitope domain of ORF2 of swine hepatitis E（SHEV) in E.coli. According to the amino acid sequence of SHEV DQ1 strain in GenBank, N-terminal major epitope domain in ORF2 was primarily mapped upon analysis of bioinformatic software. A pair of specific primer was designed to amplify the fragment encoding the major epitope domain. This fragment was cloned into the multiple cloning sites of pET30a (+) vector after subjecting to restriction endonuclease digestion. The recombinant plasmid was designated pET30a-ORF2N and was transformed into E.coli BL21 competent cells. Expression of recombinant protein was induced by addition of final concentration of 1.0 mmol/L IPTG under 37 ℃ condition. The length of RT-PCR product of interest is 424 base pairs. The size of recombinant amino acid deduced is 21.8 ku in theory. The result of Western blotting showed that the recombinant protein specifically reacted with serum against SHEV, suggesting the recombinant protein had wonderful specificity. N-terminal major epitope domain of ORF2 of SHEV was expressed successfully in E. coli and showed specificity. It can be used as antigen in diagnostic assay for measuring antibodies against SHEV in clinical practice.

Development of A Polymerase Chain Reaction Assay for Detection of Haplosporidium in Shellfish

XIE Li-ji;XIE Zhi-xun;PANG Yao-shan;LIU Jia-bo;DENG Xian-wen;XIE Zhi-qin

2010, 37(1): 54-56.

Abstract ( 1412 )

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According to the gene sequences in GenBank of Haplosporidium sp, a pair of specific primers were designed for amplifying the specific fragments of Haplosporidium sp. The reaction parameters were optimized to develop the polymerase chain reaction method for detection of Haplosporidium sp. The 244 bp long specific DNA fragment of Haplosporidium sp was amplified, but not from other pathogenic such as Perkinsus sp, Marteilia refringens, Aeromonas hydrophila, Pseudomonas fluorescens, Vibrio parahaemolyticu, Vibrio Alginolyticu, Vibrio Fluvialis and Vibrio Mimicus by PCR. The sensitivity results showed that as little as 100 fg DNA of Haplosporidium sp was detected by this PCR.

Immunologic Competence of the GP4 Protein of Porcine Reproductive and Respiratory Syndrome Virus Henan Strain

ZHAO Lin;WANG Zhong-ming;XIA Ping-an;ZHANG Feng-hua;LIU Yu-song;FAN Xu;XU Hong-yun;CUI Bao-an

2010, 37(1): 57-60.

Abstract ( 1823 )

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ORF4 gene of PRRSV Henan strain was cloned. By deletion hydrophobic sequence and cloned into the prokaryotic expression vector pET-32a, positive recombinant was successfully obtained with the induction of IPTG. Analyze purificated protein by western blotting. Immunifaction thought cytorrhyctes, degenerated protein, reannealing protein. Detect titer and neutralization of the blood serum. It can be used for the further investigation on immunogenicity of GP4 protein of PRRSV.

Cloning and Prokaryotic Expression of Nucleoprotein Gene of Porcine Reproductive and Respiratory Syndrome Virus

LUO Sheng-jun;HUANG Zhong;ZHOU Xiu-rong;JIA Chun-ling;YUAN Jie;WEI Wen-kang

2010, 37(1): 61-64.

Abstract ( 1430 )

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N gene fragment was amplified from PRRSV of cell RNA by RT-PCR and expressed it by prokaryotic expression vector pET-28a(+). A recombinant plasmid pET-28a-N was constructed and identified by restriction endonuclease digestion, then transformed into the E.coli BL21(DE3) plysS. The PRRSV N gene was expressed as a recombinant fusion protein and detected by SDS-PAGE and Western blotting. N gene fragment of 372 bp was amplified by RT-PCR, the recombinant bacteria was induced with IPTG and the expression protein was analyzed by SDS-PAGE. The results showed that bacteria contained the positive plasmid was induced to express with 1 mmol/L IPTG and 4 hours. A unique band was detected with the molecular mass of approximately 18 ku by SDS-PAGE. By analysis of Western blotting, the expressed production was reactive with the positive swine serum of PRRSV. PRRSV N gene was amplified and recombinant prokaryotic expression plasmid was constructed successfully. This showed that the high efficient expression of N protein was obtained which system of prokaryotic and eukaryotic expression. Using target protein after purified as coating antigen，an indirect ELISA was developed for detecting the anti-N antibody in the PRRSV serum by exploring the concentration of coating antigen and dilution degree of serum. 

Preparation and Initial Identification of Anti-IBV Whole Virion Polyclonal Antibody

WANG Heng;LIU Bo-qi;YIN Hang;TANG Li;REN Xiao-feng;LI Guang-xing

2010, 37(1): 65-68.

Abstract ( 1575 )

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The biological activities of an anti-IBV whole virion antibody, which was obtained from the serum of rabbit by injected the commercial vaccinal strain H120 IBV, was detected by immunohistochemistry (IHC), immunofluorence technic and Western blotting assay, and the antibody titer was analysised by agar diffusion reaction and ELISA. The results revealed that this polyclonal antibody had a high antibody titer and could specifically bind with different IBV related antigens such as pET30a-IBV S1 recombinant protein, expressing product of pcDNA3.1-IBV S1 and Beaudette strain IBV.

Development of a Real-time RT-PCR Assay for Quantificationof Rabbit GAPDH Gene

GAO B;YANG Xiao-nong;YU Xue-hui;LUO Wei;HUANG He;

2010, 37(1): 69-73.

Abstract ( 1443 )

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According to the published GAPDH gene’s CDS sequence in GenBank, a pair of primers was designed and synthesized. Through optimization of react system and conditions, the method for detection of GAPDH gene of rabbit by SYBR Green Ⅰreal-time RT-PCR was established successfully. The results show that the lowest copy number for detection of GAPDH gene with this method is 32 copies/μL. and there is a good linear relationship in a wide range from 3.2×101 to 3.2×107 copies/μL (r=0.999). The coefficient of variation (CV) of 5 different concentration of positive plasmids is 1.67% to 4.73% and 2.66% to 8.74% in intra-assay and in inter-assay respectively. The method established in this paper has the advantages of rapidity,high sensibility, high throughput and good repeatability, which provides a methodological basis for quantitative analysis on function gene of rabbit and the expression of some disease gene in rabbit when GAPDH gene is taken as a reference gene.

Isolation and Identification and Evolution Analysisof Lactobacillus johnsonii in Chicken

ZHANG Feng-hua;XIA Ping-an;CUI Bao-an;ZHANG Hong-ying;GUO Guo-fu

2010, 37(1): 74-76.

Abstract ( 1325 )

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A Lactobacillus was isolated from three days chicken’s intestinal tract. Then, the 16S rRNA gene was amplified and analyzed by PCR method, and the PCR product was cloned and sequenced. A sequence of 1340 base pair (bp) was obtained from the bacterium, and it was published on the GenBank, the location is EU290749. The sequence homology and evolution relationship between the isolated bacterium and other Lactobacillus were analyzed, its sequence homology was 99.6% with the Lactobacillus johnsonii (AB295648) that was published on the GenBank. The isolated bacterium was identified as chicken Lactobacillus johnsonii, it was named HN/0711. It may be the suitable strain of microeco praeparatum.

Research Advances in Biochip Technology

RAO Bao;XIAO Song-yun;LI Wen-gang;YANG Hui-min;WU Gao-feng;CHANG Li-li

2010, 37(1): 77-79.

Abstract ( 1368 )

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Biochip utilizes micro-biochemical analysis system built on the chip surface of micro-electronics and micro-processing technique. It process quickly and analyses sensitively cell, DNA, protein, tissue, saccharides and other organisms, the biochip technology is a high and new technique developed in recent years. Biochip includes genechip, proteinchip, tissue microarray and so on. This paper summarizes the research progress and application.

Research Progress of Porcine miRNAs

LI Hong-yi;ZHANG Yong-liang;XI Qian-yun;JIANG Qing-yan;SHU Gang

2010, 37(1): 80-83.

Abstract ( 1312 )

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miRNA is a class of non-coding small RNA about 18 to 24 nt in length，which regulates the expression of message RNA specifically. As pork has been one of the most important commodity，people pay more attention to the regulation of porcine development and meat quality，especially the recent developed miRNA. So far，only 78 porcine miRNA has been reported，the overview was composed according to the those reported miRNA，especially the muscle and fat relative ones.

The Gene Expression Mechanism of Cold Resistance in Animals

TU Yun-jie;GENG Zhao-yu;CHEN Guo-hong;SU Yi-jun

2010, 37(1): 84-87.

Abstract ( 1356 )

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Cold stress can cause performance descends，induce various diseases，even result in death in poultries. Studying the poultry cold stress mechanism and investigating scientific stress monitor method，which focus on the body biochemical indicator in cold stress reaction，have important meaning to elevate preventative level of stress and ensure livestock farming development. The gene expression mechanisms of cold resistance in animals including the functional mechanism of antifreeze proteins （AFP），heat shock protein（HSP70），c-fos and iNOS genes are summarized in this paper.

The Bacterial Carriers and Their Applications in the Development of Vaccines against Parasites

PENG Gao-hui;YAN Hai-kuo;LIU Miao-miao;ZHU Xing-quan;YUAN Zi-guo

2010, 37(1): 88-90.

Abstract ( 1436 )

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With the development of recombinant DNA technology, rapid progress has been made in genetically engineered vaccines. Live recombinant bacteria can induce both mucosal and systemic immune responses against heterologous antigens, which are easy to vaccinate, safe and cost effective. In recent years, using live bacteria as vaccine vectors are becoming hot topic in the field of vaccine studies and development. In this review, the bacterial carriers and their application in the development of vaccines against parasites were discussed, which provide a basis for further studies and development of vaccines against parasites.

The Progress of Sex Determination of Bovine Early Embryos by PCR

JIANG Xi-chun;CHENG Guang-long;ZHAO Hui-ling

2010, 37(1): 91-94.

Abstract ( 1275 )

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The current study analyzed the mechanism of sex determination by PCR method for bovine early embryos, and summarized the progress of related techniques in the past decade, including the sampling of early embryos, the extraction of cellular DNA, the design of primers, optimization of amplified condition, cryopreservation of sexed embryos and detection of product, and then evaluated the prospects of applying PCR technique in sex determination for bovine early embryos.

Genetic Diversity and Population Structure in Nine Chinese Indigenous Goat Breeds

FAN Rui;WANG Zhi-gang; LIU Chou-sheng; LUO Jun; ZHANG Gui-xiang;HAN Xu; HU Zhi-shuai; XU Li-mei

2010, 37(1): 95-99.

Abstract ( 1445 )

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Nine Chinese indigenous goat breeds and Boer were genotyped using 16 microsatellite markers recommended by the FAO and ISAG. 15 of the 16 microsatellite markers which had more than four alleles were used to analyze the genetic variation within the population and the genetic relationship and structure among the populations. A total of 159 microsatellite alleles were detected. Mean F-statistic (0.105) for nine indigenous breeds indicated that 89.5% of the variation was detected within the breeds and 10.5% of the genetic variation existed among the indigenous breeds. The UPGMA phylogenetic tree was constructed based on Da genetic distance and three clusters was obtained. The Boer separated from the indigenous breeds and formed one cluster. The indigenous goat breeds were divided into two clusters. The analysis of the population for the ten breeds showed they were separated into four clusters. The results showed that the population relationship is nearly the same to the UPGMA phylogenetic tree. 

Relationships between Genetic Polymorphisms of Intron 9 and Exon 10 of Prolactin Rceptor Gene and Litter Size of Sheep

CHEN Yong;LUO Qiu-jiang;YANG Ju-qing;YANG Feng-yun;ZHANG Ya-jun;LI Deng-zhong; ZHU Wen-yuan

2010, 37(1): 100-106.

Abstract ( 1477 )

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In the present research, the genetic polymorphisms of intron 9 and exon 10 of prolactin receptor (PRLR) gene of eight breeds or strains sheep, which including Small Tail Han, Poll Dorset, Suffolk, Germany Mutton Merino and Chinese Merino (Xinjiang type) (including Prolific strain, Meat strain and Large Frame strain), were detected by polymerase chain reaction- single strand conformation polymorphism（PCR-SSCP） and the relationship between genotypes and average litter size were analyzed. The results as follows：①There was a C→T transition at 259 in intron 9 of sheep PRLR gene. In Small Tail Han, the average litter size of BB genotype was 0.81 and 0.87 more than that of AA and AB genotypes respectively (P<0.05). ②There was a G→A transition at 304 in exon 10 of PRLR gene and this mutation cause an amino acid residue change of Glu→Lys at No. 387 of PRLR. Further research showed that average litter size of Chinese Merino (Xinjiang type) Prolific strain ewes with AB genotype was 0.58 and 0.80 more than that of ewes with AA and BB genotypes respectively (P<0.05). ③Moreover, three mutations (571G→A，585C→G and 606C→T) were identified in exon 10 of PRLR gene. The former two mutations resulted in two amino acid residues change of Ala→Thr and Ser→Arg at No. 476 and 480 of PRLR. In Small Tail Han, the average litter size of AB genotype of the 571G→A mutation increased 0.8 than that of AA genotype (P<0.05). These results preliminarily indicated that the prolactin receptor locus was either a major gene that influences the prolificacy in Small Tail Han sheep and Chinese Merino (Xinjiang type) prolific strain sheep or was in close linkage with such a gene.

The Measurement Indices and Influencing Factors of Microsatellite Polymorphisms

QIAO Li-ying;YUAN Ya-nan

2010, 37(1): 107-111.

Abstract ( 1316 )

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The principle, technique and protocols, measurement indices, influencing factors and resolve ways of domestic diversity analysis by microsatellite polymorphisms were briefly discussed in this paper. Furthermore, the progresses and application of microsatellite polymorphisms to researching sheep and goats’genetic diversity were summarized with the purpose of facilitating future study of sheep and goats breeding.

Effects of Calcium Ionophore and Caffeine on Ovine Sperm Capacitation in vitro

LIU Lie-qin;GUO Yong;LIU Yun-hai;XUE Jun-zhong;ZHANG Hong;LI Xiao-yan;NI He-min

2010, 37(1): 112-115.

Abstract ( 1342 )

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It was aimed to investigate the effects of different concentrations (0.05,0.1,0.2,0.5,1,5,10 μmol/L) of calcium ionophore (A23187,IA) and 2 mmol/L caffeine on the ovine sperm capacitation in vitro.The results showed that the higher rate of capacitation in vitro， lower the rate of acrosome reaction(AR), and the lower sperm motilily were induced by the high concentration of IA, but the dead sperm could be increased. The capacitation and AR of ovine sperm in vitro were not affected even delaying time. Caffeine could promote IA to induce AR. Collectively, this might be recommol/Lended that it be applicable for improving ovine capacitation in vitro with 0.05 to 0.2 μmol/L IA for 1 min. 

Progress in the Study on Correlative Gene of Fatness Traits in Pigs

LI Pei-pei;GE Xin;WANG Jian-hua;ZHANG Bao-xun;JIANG Ke;QU Ming-xiang

2010, 37(1): 116-119.

Abstract ( 1358 )

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With the improved requirements of pork qualities, the fat traits in pork has been a research focus. This paper mainly summarizes the progress on several candidate genes of pork’s fatness traits, which includes fatty acid-binding protein (FABP) gene, lipoprotein lipase (LPL) gene, hormone sensitive lipase (HSL) gene, diacylglycerol acyltransferase 1 (DGAT1) gene, uncoupling protein 3 (UCP3) gene, obesity (Ob) gene, etc, and supplyed some basis data to the improvement of fatness traits in pigs.

Molecular Regulation of Bovine Polymorphonuclear Neutrophil Leukocytes (PMN) Apoptosis and Immunity of Mammary Gland

YAO Xue-ping;CAO Sui-zhong;YANG De-ying;LIU Zong-ping

2010, 37(1): 123-122.

Abstract ( 1247 )

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Polymorphonuclear neutrophil leukocytes (PMN) are the first line of immunity against most pathogens that infect cattle. PMN apoptosis plays a key role in resolution of the inflammatory response in cows. This review focused on the molecular regulation mechanisms, functions and biological significance of apoptosis on immunity of the mammary gland of cows．

High Expression of Interferon-beta from Bovine in E.coli and its Antiviral Activity Analysis

LIU Ying;LIU Hua-lan;YANG Li-guo;CHEN Huan-chun;GUO Ai-zhen;

2010, 37(1): 123-127.

Abstract ( 1666 )

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Total RNA was isolated from bovine peripheral blood lymphocytes, which were stimulated with PHA. Then the bovine IFN-β cDNA was amplified by reverse transcription chain reaction. The amplified fragment was cloned into vector pMD18-T and then sequenced．The result indicated that the cloned gene was mature bovineIFN-β gene, which had the identities of 100% with the BoIFN-β gene published in the GenBank. Subcloned into the pET32a (+) expression vector and expressed in E.coli host BL21 with IPTG induction. To express the product of His-Tag fusion protein existed in the form of the expression of the total bacterial proteins account for about 40%. By nickel affinity chromatography purification of proteins, and using VSV-MDBK/IBRV cell system analysis its biological activity, reorganization BoIFN-β anti-virus activeness distinction recombinant BoIFN-β antiviral activity were about 3.16×10⁵ U/mL, 7.5×10⁴ U/mL. The result indicates that the reorganization BoIFN-β specificity is good, moreover anti-viral activeness is quite stable. This research to develop and develops the reorganization BoIFN-β class biological preparations research and development to lay the foundation.

Main Virulence Factors and Pathopoiesis Mechanism of Listeria monocytogenes

CUI Huan-zhong;QIAO Li-qiao;WANG Yi-chong

2010, 37(1): 128-133.

Abstract ( 1567 )

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Listeria monocytogenes (LM) is an important food-borne pathogen for animal and human’s listeriosis. The pathogenicity of LM is closely correlated with virulence factors. Studying the virulence factors has important meaning to understand the pathogenic mechanism thoroughly and to prevent and cure listeriosis efficiently. The article reviewed the virulence factors （LLO, ActA,PLC,Inl,p60, Ami, PrfA，VirR，and so on）and the pathogenic mechanism of LM. 

Isolation, Identification and Medicine Sensitivity Test of Haemophilus parasuis

XUE Guo-cong;XU Cheng-gang;TU Yu-rong;CAI Shao-xin;YANG Yan-lin;CAI Li-li; ZHENG Qiao-wei;ZHANG Jian-min;LIAO Ming;REN Tao

2010, 37(1): 134-137.

Abstract ( 1519 )

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Haemophilus parasuis were isolated in TSA agar medium by detecting its bacteria observation, cultural characteristics and biochemistry from infected pigs from 15 farms of Guangdong province of China, which suspected to be porcine Haemophilus parasuis. The specific PCR assays amplifying the genes of Haemophilus parasuis encoding 16S small subunit ribosomal RNA. The PCR-product of 821 bp fragments was observed with the clinical isolates. Combining with bacteria observation, culture characteristics and biochemistry, all results showed that seven isolates of Haemophilus parasuis were successful identified. Drug sensitive tests showed that isolations were sensitive to Ampicillin, Polymyxin B and Gentamycin.

Reverse Genetic Technique and its Application in Foot-and-mouth Disease Virus

ZHU Xiu-mei;MU Hong-tao;HUANG An-cang;HUANG Yin-jun;WEI Guang-sen

2010, 37(1): 138-140.

Abstract ( 1518 )

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Reverse genetics is a new developed molecular biology technique. It can run a variety of modify and transform on DNA level for virus genome，and then these genetic manipulation effect is judged by phenotype change of rescued virus. So it can be used to research the expression and regulation mechanism of virus genome and molecular mechanism of virus pathogenicity. The technique has been applied in foot-and-mouth disease virus research，the purpose, methods and applications were also reviewed.

Experimental Study on the Toxicology of Gentamicin and Lincomycin Injection

JIA Hai-yan;LI Yan-hua;LI Ji-chang;GUO Bin

2010, 37(1): 141-144.

Abstract ( 1698 )

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To evaluated the safety of Gentamicin and Lincomycin injection, by acute toxicity test, hemolysis test, local vascular, muscleand skin irritation tests and systemic allergy test in guinea, the safety of Gentamicin and Lincomycin injection was inspected. LD₅₀ of Gentamicin and Lincomycin injection by intramuscrular adminstration in mice was 4.989 mL/kg, hemolysis or coacervation in the red blood cells in rabbts did not occur for 0.1 to 0.5 mL of the injection in 4 h; Pathological changs such as hyperaemia,edema or hemorrhage were not examined in the veins where were injected intravenousiy and their surrounding tissues the hyperaemia ranged below 0.5 cm×1.0 cm on the intramuscular injection site, the ifference value between the maximum and the minimum order of reaction in quadriceps was zero， moreover the sum of them was less than 10;Double dose of the clinical doage of the injection was smeared on the rabbit skin, obvious pathological changes were not observed after the drug withdrawal for 1，24，48 and 72 h; Anaphylactic reaction was not observed in the guinea pigs after they were first sensibilized for 14 and 21 d. The results showed that, under this experimental condition, Gentamicin and Lincomycin injection was safe.

Progress on Virulence-associated Factor of Swine Streptococcus suis Type 2

GUO Li-li;XU Cheng-gang;ZHANG Jian-min;LI Jing-yi;CHEN Ji-dang;LIAO Ming

2010, 37(1): 145-148.

Abstract ( 1495 )

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At present，swine Streptococcus suis，a more important zoonotic pathogenic bacteria，has a serious effect on the development of the pig farming industry and human health. The bacteria has thirty-five serotypes，swine Streptococcus suis type 2(SS2) is considered the most virulent serotype, and is pandemic in the worldwide. The major virulence factors play an important roal on pathogenicity. But its virulence factors are the key to effectively combat disease. Then this article mainly summarizes the virulence factors and several virulence-associated factors，which supply the scientific substructure and reference for effective prevention and treatment of this disease.

Identification of Toxigenic Pasteurella multocida

LI Wei-jie;ZHAO Yun;DU Xin-bo;KANG Kai;CHEN Min

2010, 37(1): 149-151.

Abstract ( 1509 )

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Colony multiplex PCR assays for species，capsular type and dermonecrotic toxin were conducted to identify twenty-eight Pasteurella multocida strains conserved. The results showed that strains C51-6，M-4 and P-2237 were toxigenic Pasteurella multocida，strains C51-6 and P-2237 were serotype D and strain M-4 belonged to serotype A. At the same time，Staphylococcal inhibitory test，neutral acriflavine precipitation and guinea pig skin test were conducted to verify the PCR assays. Based on the ability to utilize mannitol，dulcitol，sorbitol，trehalose and by the production of ornithine decarboxylase，the Pm strain C51-6，M-4 and P-2237 were identified as Pm subsp. multocida. 

Study and Application on Drug Screening and Programmed Prevention Mode of Parasitic Diseases in House-feed Sheep and Goat

ZHAO Jin-shan;WANG Jun-wei;CHENG Ming;JIANG Ke;WANG Wen-xin;DAI Zheng-hao

2010, 37(1): 152-154.

Abstract ( 1344 )

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In this study, Niclofolan, Compound Doxycycline Hyclate and Florfenicol Infection, Albendazole, Phoxim Pour-on Solution, Ivermectin and Chloroquine were selected to experiment in vivo and in vitro of sheep and goat, which based on main varaties, epidemiology etc of parasites in Qingdao, these druggery were selected with sensitivity, programmed prevention mode of parasitic diseases which suitabled for house-feed sheep farm of Qingdao was developed.

Isolation，Identification and Drug Susceptibility of Streptococcus suis 2 from Large-scale Pig Farms in Henan Province

XU Yin-di;WANG Zhi-fang;ZHU Wen-hao;JIAO Hai-yan;LIANG Yue;GUO Cheng-liu

2010, 37(1): 155-157.

Abstract ( 1375 )

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Gram postitive coccobacilli were isolated from 291 samples of 113 farms in Henan province which had high fever syndrome from 2006 to 2009. These strains were identified by culture characteristics，biochemical test，SS and SS2 PCR assay. The results showed that there were 125 strains Streptococcus suis and 35 strains SS2 among these 125 SS. Mice virulence and drug susceptibility test results showed that these 35 SS2 strains were lethal to mice and sensetive to Cefotaxime，Cefazolin，Ceftazidime，Amoxicillin，Chloramphenicol，Ciprofoxacin and Ofloxacin.

Pathogenesis Progression of Porcine Reproductive and Respiratory Syndrome Virus

LIU Ping;CHEN Miao-miao;QIAO Li-ping;WANG Fan;LIU Xue-rong;MU Ke-bin;HUANG Yin-jun 

2010, 37(1): 158-160.

Abstract ( 1560 )

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Porcine reproductive and respiratory syndrome（PRRS），caused by porcine reproductive and respiratory syndrome virus（PRRSV），is a serious infection disease of swine. PRRS has already been one of the most economically important disease of global swine industry. This paper reviewed pathogenesis of the PRRSV to guide the study of diagnostic technique， mechanism of immunity， research of new vaccine and prevention of PRRS.

Antibody Detected and Immune Effect Analysis of Main PigDiseases in Some Regions of Guangxi

WEI Xian-kai;ZOU Lian-bin;QU Su-jie;LIANG Yuan;ZHAO Guo-ming;HE Yi-jian;MO Sheng-lan;ZHENG Lie-feng;GUO Jian-gang

2010, 37(1): 161-163.

Abstract ( 1364 )

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393 pig serum samples were collected randomly from 33 farms in 5 regions，these pigs had been vaccinated PRRSV，CSFV，PRV vaccine. The antibody level of these serum samples were detected with immune colloidal gold technique. The results showed that the highest ratio of positive antibody in different scale of farms was CSF(75.06%)，follow by the PR(71.25%)，the lowest was PRRS(42.24%). Antibody level was various from different scale of farms，antibody level in large scale farms was not necessary higher than the dispersive. Compared the antibody level which vaccinated by different kinds of vaccine，there were no significant difference in the following respectively: inactivated and attenuated PRRSV vaccine，rabbit origin or cell origin CSFV vaccine，homemade and imported PRV vaccine.

Subchronic Toxic Experiment of Oxytetracycline Herb Residues Removed the Titer

SUN Hong-li; LIU Zhi-qin; XU Hong-xin; YANG Bao-zhu; REN Guo-qiang

2010, 37(1): 164-165.

Abstract ( 1300 )

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The subchronic toxic of oxytetracycline removed the titer was tested by feeding mouse. Whether toxic substance is produced in the process of treating the oxytetracycline herb residues was studied. The results showed that the organ coefficient of mouse fed the worked herb residue was similar to the mouse fed the unworked herb residue (P>0.05). The indexes of worked herb residue are identical with that of unworked herb residue and it decreases the residues titer greatly. 

Isolation and Drug Resistance Monitoring of Duck Streptococcus

ZHAO Rui-hong;ZHANG Dan-jun;PAN Xiao-cheng;CHENG Bao-yan;HU Xiao-miao

2010, 37(1): 169-170.

Abstract ( 1289 )

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This experiment isolated 19 strains Streptococcus successfully from the internal organs of disease duck used blood agar and nutrient agar culture medium. We can see gram positive coccus array like catenuliform through microscope after Gram staining. Biochemical test and animal inoculation test showed that the isolated bacteria are pathogenic to duck. Drug sensitive test showed that the drug resistance of isolated streptococcus is very high. Twelve strains Streptococcus is low sensitive or not sensitive to Amikacin, Cefatriaxone, Ofloxacin so on fourteen medicaments except seven strain Streptococcus is middling sensitive to Amikacin. This indicates that the enhancing drug resistance of duck streptococcus makes it hard to prevention and cure of duck Streptococcus disease.

2010, 37(1): 178-179.

Abstract ( 1424 )

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Development and Application of Indirect ELISA for the Detection of Serum Antibodies against Riemerella anatipestifer in Duck

LI Fu-xiang;YANG Bin;CHANG Zhi-shun;WANG Chuan-yu;YAO Jun;LIU Yan-li;LI Hua-chun

2010, 37(1): 202-205.

Abstract ( 1621 )

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Indirect ELISA was developed and applied for detection of serum antibodies against serotype 1(Yunnan strain)Riemerella anatipestifer (RA) in ducks, using ultrasonic RA as the coating antigen. Dilution method shew the optimum HRP-Goat Anti-Duck IgG dilution was 1∶3500, chessboard test shew the optimum coating antigen concentration was 2.7 mg/mL,the optimum dilution of serum samples was 1∶300 and the cut-off value of negative serum was 0.381.The interruption test shew the reactions between antiserum of serotype 1 RA and the coating antigen could be interrupted by specific type 1 RA, but could not be interruped by E.coli serotype O78 and serotype O132 strains(no cross-reaction).Repetition test shew the variant coefficients of the D value within assay and between assays less than 0.246 and 0.889 respectively. An interruption test and repetition test showed that the ELISA assay had good specificity and reproducibility to detect serum antibodies against serotype 1 RA in ducks.

Effects of Echinacea Herb P.E on Micronucleus in Mice

ZHAO Dan;CHEN Xiu-yun;SUN Yu;TANG Ren-xiang;LIU Ren-jie;BU Shi-jin

2010, 37(1): 208-210.

Abstract ( 1456 )

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50 Kunming mice were randomly divided into five groups. Echinacea Herb P.E was orally given at a dose of 2.5, 1.25 and 0.625 g/kg body weight, respectively. The bone-narrow micronucleus test was undertaken according to the routine method. With a method of two times administration at intervals of 30 h,the mice were killed by cervical dislocation 24 h after oral administration, the permillage of micronucleus cells among polychromatic erythrocytes (PCE) were numbered. The result showed that micronucleus test were not significantly different from the control group, and suggested that Echinacea Herb P.E is no genotoxicity.

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