2 single nucleotide polymorphism (SNP) sites of the 5′ UTR of insulin-like growth factor I (IGF-I) gene were detected among Tibetan chicken and Recessive White Chicken using polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP). The results displayed that there were HinfI and PstI polymorphisms in the 5′ UTR of IGF-I gene and base pair mutations A→C and C→T took place by DNA sequencing, respectively. 3 genotypes and 8 haplotype combinations were present within 2 breeds. The results of Chi-square tests of haplotype combinations showed that there were significant differences at 0.01 level between two breeds. The outcomes of analysis of variance proved that breeds had the significant effect on 17 growth and development traits (P＜0.05 or P＜0.01), sex observably affected 6 growth and development traits (P＜0.05 or P＜0.01), and genotypes/haplotypes combination had remarkable influence on 5 growth and development traits (body weight at 1 day, body weight and shank girth at 2 weeks of age, body length at 7 weeks of age and shank girth at 16 weeks of age) (P＜0.05 or P＜0.01). At the same time, the interactions between genotypes/haplotype combinations and breeds had also influence on some of chicken growth and development traits. In addition, on the mean value of chicken body weight at 1 day and that of body length at 7 weeks of age, genotype CT was significant higher than genotype CC and TT at 0.05 or 0.01 levels, and on the average value of chicken body weight at 1 day, that of body weight and shank girth at 2 weeks of age and that of shank girth at 16 weeks of age, Haplotype combination A+T/C+T was significant higher than others at 0.05 or 0.01 levels.

Adenosine monophosphate deaminase 1 (AMPD1) gene was studied as a candidate gene of inosine monophosphate (IMP) content of Beijing fatty chickens. According to the sequence of AMPD1 gene of human and rat, primers were designed to amplify the AMPD1 gene of chicken. Based on the clone sequencing, single nucleotide polymorphism of AMPD1 gene was detected in 130 Beijing fatty chickens by PCR-RF-SSCP. The results showed that three alleles (A,B,C) and six genotypes (AA,AB,AC,BB,BC,CC) were found in Beijing fatty chickens. Frequencies of alleles A, B, C were 0.180 8, 0.661 5, 0.157 7, respectively. Frequencies of genotypes AA, AB, AC, BB, BC, CC were 0.076 9, 0.176 9, 0.030 8, 0461 5, 0.223 1, 0.030 8,respectively. The least squares means of IMP content for AC and BB were higher than that for AA (P<0.05), and there were no significant differences between other genotypes (P>0.05).

A cDNA library was constructed with mRNA from the 105th day’s fetal skin of cashmere goat. 846 ESTs were analyzed, with the average length of 443.2 bp. Five hundred and twenty five known genes were proved, and they are 18 cell divisions, 65 cell signals, 73 cell constructions, 21 cell defenses, 126 gene/protein expresses, 69 metabolism and 153 unclassified. Wnt-4, Bmpr-Ib, ASIP gene and Ectodysplasin A gene were discovered， which are important candidate genes for the hair follicle development.

Three pairs of Y chromosome specific primers for different gene in cattle were designed and applied to check genomic DNA of cattle, yak, sheep, goat, pig, mouse respectively. The amplification results of all three pairs of primers were obtained from genomic DNA of cattle and yak, only one amplification results of sex determine gene was obtained from sheep genomic DNA, and no amplification result was checked in the other species. After all the amplification products were cloned and sequenced, the sequences were blast with DNAMAN between cattle and yak or sheep. The identities of three sequences are more than 99% between cattle and yak, the identity of segment of sex determination gene is 94% between cattle and sheep.

The dual expression vector pVITRO-FSHαβ was successfully constructed on the basis of cloning of goat FSH α & β and transient expression. The plasmids were transfected into Chinese Hamster Ovary (CHO) cells, and stable expressing cells were acquired by screening of Hygromycin B. These results would make preparations for producing recombinant hormone.

In vitro-produced bovine parthenogenetic embryos were cultured with Vitamin E ,Vitamin E and Vitamin C in CR1aa culture medium at 38.5 ℃, 5%CO² and high-humidity air. Meanwhile, fluorescence dye Hoechst 33342 was used to stain the blastocyst, so as to calculate the total nuclei number of per blastocyst. The result shows that when added 100 μmol/L Vitamin E to the culture medium, the development rate of expanded blastocyst and the total number of blastocyst raised distinctly compare to the control (P<0.01). When added Vitamin E and Vitamin C to the medium, the number of early blastocyst, expanded blastocyst, hatched blastocyst and the total cell number of blastocyst were lower to the Vitamin E treatment.

The present study was conducted to determine the effect of feeding diets with whole crop rice brittle culm-1 (WCR) on growth performance, nutrient digestibility, carcass traits and meat quality of growingfinishing pigs. Fiftyfour female pigs (Duroc), with initial mean weight of about 29 kg, were divided into three equal groups and the dietaries were infused with 0, 10% or 20% WCR, respectively. During the grower period, the pigs fed 10% or 20% WCR diet were not significantly different in feed intake, average daily gain and gain-to-feed ratio from those on the control. During the finisher period, the feed intake and average daily gain of pigs fed 20% WCR diet was significantly lower than that of animals fed the control (P<0.01), while the pigs fed 10% WCR diet were not significantly different in these two parameters from those on the control, nor was there significant difference in gain-to-feed ratio among pigs fed different diets. The digestibilities of nutrients in 10% or 20% WCR diets were lower than those of the control during grower period, while little difference was found in the digestibility of nutrients except NDF between WCR and control diets during finisher period. The dressing percentage and mean backfat thickness of pigs fed 10% or 20% WCR diet were lower than those of pigs on the control, whereas the lean percentage and loin eye area were superior in the WCR diets to those in the control, though the differences were not statistically significant. The longissimus muscles of pigs fed 20% WCR diet had greater protein and lower fat content (P<0.05) and higher 24 h postmortem pH value (P<0.01) than that of control. There was no significant difference in drip loss and marbling and color scores for longissimus muscles among pigs fed different diets.

16 of 30 days’ Romney Marsh×Xinjiang Merino male lambs，at the body weight of 6~10 kg，with rumen fistula，were divided into 4 groups，and were fed 4 diets with DE/CP(MJ/%) of 11.91/20，1191/14，8.15/20 and 8.15/14 respectively，to study the development of the digestion and metabolism in rumen. At the last 3 days of every 15 days，the rumen fluid were sampled at 0，1，3，6 and 9 hours after feeding during the age of 30~150 days for assaying. The results showed that the pH， NH3N，propionic acid，and acetic+propionic+butyric acids concentrations in rumen fluid were affected by dietary nutrient level，increased with the age， and were stable after 100 days’ age. The effect of age and dietary nutrient level on acetic acid in rumen fluid was less. The rumen fluid volume was increased almost by 15 times in 120 days，as well was related with nutrient level of diet. This study elites that the development of the digestion and metabolism of rumen is progressive， which is mainly dependent on the increase of rumen fluid volume，and at 100 days age it presents a stable tendency.

Seventy-two Laiwu Black and Xin Laiwu pigs (weight of 40~90 kg) were used to study the development changes of fatty enzymatic activities and their association with intramuscular fat (IMF), backfat thickness (BFT). Results showed as follows: (1) With gaining of body weight, the activity of ICDH was significantly higher than that of MDH in muscle(P<0.01). The activity of ICDH reached a peak at 60~70 kg of weight and decreased thereafter. MDH had no significant development changes. The activity of HSL fell firstly and reached a peak at 70~80 kg of weight. (2) The activity of MDH was significantly higher than that of ICDH in backfat(P<0.01). Both MDH and ICDH activities, to some extent, decreased firstly and then increased. The activity of HSL was stable at 40~50 kg and then increased gradually. (3) The activity of ICDH was significantly higher than that of MDH in liver. The activities of ICDH and MDH increased firstly and tended to stabilize after 60 kg. The activity of HSL had no significant development changes. (4) The activities of MDH, HSL in muscle were highly correlated to IMF content (P<0.01). MDH, ICDH in liver positively correlated with IMF, and ICDH also with BFT(P<0.01). The activities of MDH, HSL in backfat were strongly correlated to BFT (P<0.01). It implied that there was not only some immanent contact between the depositions of IMF and BFT via liver, but also some independence from each other. By selecting fatty enzymatic activity in muscle, it is possible to improve IMF content but not to change BFT.

To explore the effects of cadmium on female procreation function, prepubertal female mice were injected with several dose cadmium chloride after subcutaneous injecting with PMSG 48 hours. The apoptosis of granulose cells were detected by TUNEL and agarose gel electrophoresis; the expression of bax gene was detected by immunohistochemical method. The results indicated that the apoptosis percent of granulose cells in high-,midst- and low-dose groups were ( 13.887 1±1.329 9)% , (10.757 3±1.229 3)%, (10020 8±0.983 7)% respectively after 48 hours,and were all higher than that of control group significantly(0.637 8±0.387 2, P<0.01).Apoptosis percent of GCs rised obviously after 96 hours’treatment in control group,there is a significant difference between high-dose group and others ,but midst-and low-dose groups are similar with control group. DNA ladder was found only in 96 hours’ treatment high-dose group by agarose gel electrophoresis. Expression of bax in 48 hours’ and 96 hours’ is higher than control group respectively. These results showed that cadmium can accelerate the apoptosis of GCs and is related with dose; TUNEL is better than gel electrophoresis in detection of apoptosis. No ladder was revealed by gel eletrophoresis if the apoptosis rate was low enongh（＜20%).There is a positive relationship between apoptosis of GCs and expression of bax in some extent.

To study the distribution and decrepitude change of nitric oxide synthase (NOS) positive neurons in rabbit brain, the samples from different ages were collected and were observed by NADPH diaphorase(NADPH-d)histochemical technique and microscope. The results showed that NADPH-d positive neurons distributed in wide areas of rabbit brain including cerebellum,cerebral cortex,hypothalamus, mesencephalon and pons. A few of positive cells could be seen in the medulla oblongata. NADPH-d positive neurons appears blue and its karyon hasn’t pigmentation, its dendriate and axon’s pigmentation are clear. The results indicate that NO may be involved in many brain fuctions.

Based on windows 2000 advanced Server and SQL Server2000 network database system，this study has been digitized almost production process factors during a intensive dairy farm progress and set up a systematic database framework that includes over 100 data tables, and finally constructed a precision farming technology Platform of Dairy Cattle Farm on Internet/Intranet by combining both frame B/C and frame C/S. With the aid of automated created stimulation data according to supposed basic parameters in advance inside the platform, some expected objectives has been tested. The result shows as follows: it can track at least five generation genealogy in terms of individual dairy cattle, and inspect inbreeding coefficient of capable born descendants，and dynamically show herd profile made up by different types of cattle，especially , it can calculate nutrients requirements of individual dairy cattle in terms of its body condition parameters and systeminside designed forecasting models, and optimize diet formulation for each and realize precision feeding of dairy herd on single livestock information. Finally, it points out the developing direction of this technology platform is to set up real three tiers framework based on component technologies and provide more large space for Web Server.

A in situ hybridization(ISH) method was used to detect low copy FMDV in infected cells, in order to illuminate FMDV multiplication site. In this study, FMDV RNA was located and detected in BHK21 cells by ISH. Cells infected experimentally with O/Akesu/58 (10⁷TCID₅₀) at 2, 4, 6, 8 and 10 hours postinoculation. Oligonucleotide sequences were chosen from the highly conversed regions of 3D gene of FMDV RNA. Probes were labeled with digoxigenin-11-dUTP at the 3'-end. Hybrids were detected by enhanced sensitive ISH detection kit Ⅱ(AP). Results showed that positive signals were found in the cytoplasm of infected cells post-inoculation, lots of viruses gathered periphery nucleus. Intensive positive signals showed in the cytoplasm of infected cells at 6, 8 and 10 hours postinoculation. So, we suggested ISH combined with enhanced sensitive ISH detection kitⅡ(AP) is a sensitive method, which was a reliable method for detection and localization of low copy viruses in infected cells. Results indicated multiplication and duplication of FMDV were in the cytoplasm of infected cells.

Fifty Peking ducks aged two months were inoculated subcutaneously with a virulent field strain of duck enteritis virus.The morphology of cells of those infected ducks was observed using electron microscopy and ultrathin section techniques.The result make it clear that the lymphocytes in spleen,thymus,bursa of Fabricus and in the lamina propria of small intestine undergo apoptosis and necrosis at the same time.At the process of the disease,the number of apoptotic cells increased steadily.The morphological changes of apoptosis including chromatin condensation and margination,appearance of vacuoles on the cytomembrane,and the formation of apoptotic bodies.The apoptosis and necrosis of lymphocytes resulted in the depletion of lymphocytes,and the apoptosis of lymphocytes may play an important role in the pathogenesis of duck viral enteritis.

For evaluating the efficacy of DNA vaccine pCAGGoptiHA5, 6 groups of 3-week-old SPF chickens were intramuscular inoculated singly and boosted with 100 μg or 10 μg of pCAGGoptiHA5 in 200 μL volume. A group of chickens were injected with 200 μL PBS as controls. Sera were collected weekly after vaccination for detecting the HI, NT and AGP antibodies. Four-weeks after the single vaccination, two weeks after the boost, all chickens were challenged with 100LD₅₀ of highly pathogenic A/Goose/GuangDong/1/96(H5N1)strain, oropharyngeal and cloacal swab specimens were collected from all chickens 3, 5 and 7 days after inoculation for titration of virus in eggs, and chickens were observed daily for disease signs and deaths for 2 weeks. Results shown that 100 μg or 10 μg pCAGGoptiHA5 boosted chickens, 100 μg pCAGGoptiHA5 priming chickens were completely protected from virus challenge (no disease sign, no virus shedding and no death).The constructed pCAGGoptiHA5 could protect chickens from lethal H5N1 virus challenge even at the low dose of 10 μg, implied the potential possibility for the commercialization of avian influenza DNA vaccine in the future.

To construct a cDNA library from adult Fasciola gigantica. Total RNA was extracted from Fasciola gigantica by Trizol regent. The first strand cDNA was synthesized and the second strand was amplified by Long-Distance PCR. The Chroma Spin-400 was applied to purify cDNA fragments longer than 400 bp. The purified fragments were ligated with λTriplEx2, then were packaged in vitro,at last the cDNA phage expression library was constructed by infecting XL1-Blue.The unamplified library capacity was 1.08×10⁶ PFU/mL, the library recombination rate was 96.6%.The titer of the amplied library was 2.41×10⁹ PFU/mL. The cDNA library of adult Fasciola gigantica has been constructed using SMART LD-PCR technology for the first time.

Total RNA were isolated from organs such as salivary glands, Malpighian tubules, ovaries dissected from partially engorged Haemaphysalis qinghaiensis. Subsequently mRNA were purified. A library of oligo(dT)-primed cDNA with added directional EcoR I/Hind III linkers was constructed from the purified mRNA.The constructed cDNA was ligated to the EcoRI/HindIII arms of the λSCREEN vector. The recombinant phage DNA was packaged by using PhageMaker packaging extracts, resulting in a primary cDNA library with a size of 2.0×10⁶ PFU. Data showed 100% of the library were recombinant and the titer of the amplified library was 8×10⁹PFU. A full-length cDNA encoding myosin light chain alkali of Haemaphysalis qinghaiensis was screened from the expression library using rabbit anti-Haemaphysalis qinghaiensis differential proteins serum. The results suggested that the cDNA expression library was constructed successfully.

The present study examined the nucleotide variation in a portion of the mitochondrial (mt) NADH dehydrogenase subunit 1 (nad1) between members of the P.decipiens complex using a PCR approach. The objective was to assess whether mtDNA sequences provide genetic markers for the identification of members of the P.decipiens complex. The results showed that sequence differences in the nad1 gene between species was 3.6% to 15%, indicating that nad1 gene sequence provides genetic markers for the identification of the 6 numbers of the P.decipiens complex. These findings provided basements for further studies of population genetics of members of P. decipiens, and for the diagnosis of diseases they caused.

The experiments on the transmission of Babesia U sp. and Theileria annulata with ticks were carried out to demonstrate that Hyalomma anatolicum anatolicum is important for bovine piroplasmosis. The results showed that Hyalomma a. anatolicum was capable of transmitting Babesia U sp. transovarially in nymphal (2/2) and adult (3/3) stages respectively, and that it also transmitted Theileria annulata transtadially in nymphal (2/2) and adult (2/2) stages, and that larval and nymphal progeny of Hylomma a. anatolicum infected by Babesia U sp.could be still infected by Theileria annulata and the nymphs (2/2) and adults (2/2) developed from them could transmit Babesia U sp. and T. annulata to the calves without changes in incubation period and clinical signs.

The total calcium and Ca²⁺in right ventricle of broiler were evaluated to study the relationship between calcium signal transduction and ascites syndrome (AS) in broiler chickens. The total calcium in right ventricle(RV) of broiler was measured by atomic absorption spectrophotometer. The result showed that the total calcium of RV with AS is significantly higher than that of broilers reared in low ambient temperature and normal temperature (P<0.01). The quantity and distribution of Ca²⁺ in right ventricle was observed by pyroantimonate precipitation. The result showed that Ca²⁺ was mainly located in cytoplasm, sarcoplasmic reticulum and mitochondrion. The quantity of Ca²⁺ in myocardium of broiler with AS was more than that of normal broiler. The increase of total calcium of RV and the precipitation of Ca²⁺ in the cardiac muscle cell of broiler with AS suggested that calcium abnormal play an important role in the development of AS.

On the basis of B-mode ultrasonic image, 40 gestational Labrador dogs were monitored by 3.5 MHz scanner on the body surface, its pregnancy and the embryo’s growth were observed, and the first time to examining out the structure of embryo outside, embryo appearance and the structure of embryo inside were recorded. The first time of measuring gestational sac, fetus heart beating and placenta out were 18 d, 23 d and 31 d, respectively. The intact skeletal system could be detected on earliest 36 d, and most viscera organs could be determined on 37 d. The fetus’ sex cannot be identificated by 3.5 MHz scanner accurately，and fetal numbers is difficult to be determined.

In order to select Chinese herbal medicine ingredient (CHMI) and develop a new-type compound CHMIs immunopotentiator, the effects of nine Chinese herbal medicinal ingredients, Chinese angelica polysaccharide (CAPS), astragalus polysaccharide (APS), isatis root polysaccharide (IRPS), epimedium polysaccharide (EPS), propolis polysaccharide (PPS), epimedium flavone (EF), propolis flavone (PF), astragalosides (AS) and ginsenoside (GS), at four concentrations on promoting splenic lymphocyte proliferation were compared in chicken. The results showed that most of CHMIs had better action, in which APS, EPS, AS and GS etc. could stimulate the lymphocyte proliferation singly or synergistically with ConA or LPS, the actions of EPS and APS were the strongest. At the same time, according to the above experimental results, one suitable concentration of each CHMI was chosen and their efficacies were further compared. It was found that the effects of EPS and GS were the best. These CHMIs would be expected as the component drugs of the new-type compound CHMIs immunopotentiator.

Avian broilers were randomly divided into 5 groups(54 birds respectively)at the age of 15 d, one was normal control group(N) which was reared at normal ambient temperature,(21±1)℃; the other four groups were reared at low ambient temperature,(11±1)℃,to induce ascites syndrome. These latter four groups were: control group(C) not given any medicine, vitamine group(V) given VC(0.5 g/L) and VE(0.1 g/L) in drinking-water, test group T1 and test group T2 given tested Chinese medicine, LGZG Tang,0.4 mL/kg bw and 0.8 mL/kg bw separately in drinking-water. The results showed that the incidence of ascites syndrome, ascites heart index(AHI),packed cell volume(PCV),red blood cell count(RBC),hemoglobin(Hb) and malondialdehyde(MDA),α-hydroxybutyrate dehydrogenase(α-HBDH) and two tansaminases ［glutamic-oxaloacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT)］ in the serum were significantly higher in the C-group than in the N-group; but the three antioxidases ［superoxide dismutase (SOD), glutathione peroxidase(GSH-Px) and catalase］ were significantly lower in the C-group than in the N-group. All the parameters mentioned above were improved in the test groups. It was revealed that LGZG Tang was able to improve the antioxidation capability of the broilers and prevent ascites syndrome.

The component changes of Chinese herb before and after fermentation were analysed by HPLC and the efficacy was compared based on their application to broiler chickens. According to chromatographical analysis,the amount of some components in fermented Chinese herb decoction was 1～4 times as much as those in non-fermented decoction, and some new absorptive peaks appeared while some specific absorptive peaks in non-fermentative herb disappeared.The results showed that HI titer of anti-ND antibody, index of immune organs and RBC level of erythrocytes in fermented group were all significantly higher(P<0.05)than those in non-fermented group;and the utility rate of fermented fodder increased obviously.It showed that some new components appeared in fermented Chinese herb decoction and its efficacy was better than non-fermented herb.

In order to amplify the partial bcl-2 gene of goose, we designed a pair of primers according to the reported chicken bcl-2 gene sequence in the GenBank, and obtained the gene by reverse transcript-polymerase chain reaction (RT-PCR). Then the gene was cloned into pMD-18T vector and identified by PCR. The nucleotide and amino acid sequences of goose partial bcl-2 gene were compared with the counterpart sequences of chicken, and the nucleotide homology was 92.49%. This is the first report of bcl-2 partial nucleotide sequence of goose.

Ciprofloxacin MIC was tested by the disc diffusion method in 25 Escherichia coli isolated from diseased livestock and fowl.There were 15 susceptible and intermediate(MIC≤1.0 mg/L) and 10 resistant(MIC≥32 mg/L) isolates.The expressing level of AcrA(an obligatory component of the AcrAB multidrug efflux system) was determined by Western-blotting, and gene mutations in gyrA and parC were determined by PCR.The results showed that the expression of AcrA protein was exceeded 170% and had gene mutations in gyrA and parC in 9 of 10 clinical isolates of Escherichia coli with high-level ciprofloxacin resistance but not in any of the 15 isolates for which the MIC was less than 1.0 mg/L.These results suggested that Topoisomerase and AcrAB efflux pump mediated highlevel fluoroquinolone-resistance in Escherichia coli.

Events related with gene frequencies variations in breeds characters, biochemical polymorphism, DNA level gene expressions were cited in given paper. Breeds are the extension parts of genetic diversity of wild animal species. When the species have been one member of eco-chain, lose of it would be reason for probably collapse of an ecosystem. A breed is one part of sustainable production chain replacing old ones by improved ones, which would be more benefit than before. Selection is the best way for improving breed traits. The gene frequencies changes caused by eco-conditions, migrations caused by imported breeds, narrow gene founds in rare breeds , and mutation, especially artificial selection are all the facts for breakaging away from gene equilibrium, also AI-insemination is one of these facts. In research for breeds-cluster analysis the above mentioned events must be taken into consideration in order to avoid leading a biased conclusion.