The objective of this paper is to provide an introduction to generalized linear mixed models and to compare of the efficiency of the GLMM and LM. The method of GLMM is described that allows genetic analyses and predicting breeding value for resistance traits. Binary and ordinal response traits are simulated in this study, and the design is mixed family of full-half sib. The logit and log linkage function are applied, respectively. The method of Fisher score is applied to calculate the parameters in generalized linear mixed models. The results showed that generalized linear mixed model can predict breeding value to a nicety. In addition, the method of GLMM has a great advantage in predicting the breeding values for resistance traits.

Nine genotypes of BoLA-DQA*exon2 allele, fifteen genotypes and three haplotypes of BoLA-DRB3* exon2 allele were observed in Chinese Holstein using PCR-SSCP. Correlations between all genotype or haplotype with the mastitis were analyzed in this study. The results showed that susceptible or resistant gene and genotype of mastitis was not observed in Chinese Holstein Cattle, DQA-B/DRB3-C and DQA-G/DRB3-F haplotypes might relate to the susceptibility of mastitis, DQB-F/DRB3-E haplotype may relate to the resistance of mastitis.

In order to study exon10 of prolactin receptor gene（PRLR）in alpacas, and to analyze the homology to other mammals, blood samples and tissue from 60 Huacaya alpacas, breed in China Alpacas Base, were taken and their genomic DNAs were extracted by the method of hydroxybenzene / chloroform, and then cloned, sequenced and submitted to GenBank（DQ206831）. The sequence analysis revealed that the alpaca prolactin receptor exon10 spaned 1 133 bp, and included 1 046 bp encoding region and 87 bp untranslated region. And the multiple sequence alignments revealed that the homology of alpacas is 80% to the other mammals ( such as cow, sheep, pig, dog, rabbit, rat and so on), the sequence of amnio acid is more than 66%, but it is only 40% to 45% to the fish at the amnio acid level.

FGF5 is a member of the FGF superfamily. It is specifically expressed in hair follicles cells and functions as a negative regulator of follicle anage. This study was designed and aimed to identify the single nucleotide polymorphisms (SNPs) of FGF5 gene by PCR-SSCP (singlestrand conformation polymorphism) in various sheep breeds including Small Tail Han sheep, Hu sheep, Tibetan sheep, Chinese Merino sheep. The 3 pairs of primers for FGF5 gene were designed based on the human and mouse genomic sequence. Two SNPs were identified in the sheep FGF5 gene by PCR-SSCP and sequencing. They were in the exon 1 region in different sheep lines. The result of these analysis indicated that the polymorphisms are due to two single point mutation G to T and C to T at base position 122 and 385, respectively. Population genetic analysis indicated that genotype frequency of the P1 SNPs in Chinese Merino sheep were quite different from other tested sheep breed. Frequency of the BB genotype was much higher in Chinese Merino sheep while the AA genotype was much lower than other breed. The genotype frequency of P1 was significantly different in the tested sheep lines (P<0.01). All the population were in Hardy-Weinberg equilibrium state. For primer 2, frequency of the EE genotype in Tibetan sheep was higher than those in the other lines, Frequency of allele E in primer 2 (P2) in Tibetan sheep was higher than that of other breed. All the population were in Hardy-Weinberg equilibrium state except Chinese Merino sheep.

Porcine fetal fibroblast cells, ear fibroblast cells and granulosa cells were used as donor in pig cloning. We compared the effects of cell cycle synchronization treatment of fetal fibroblast cells into G0/G1 by serum starvation or contact inhibition. There were no significant difference in percentage of G0/G1 between two treatments, no significant difference in percentage of G0/G1 by serum starvation for 2d and 4d, as the same when by contact inhibition. Serumstarved fetal fibroblast cells or cycling growing subconfluenced cells， round and smooth vs. rough surface cells,different types of somatic cells and fibroblast cells of different individuals were used as nuclear donor in porcine somatic cloning. The results indicated that serum starvation could not improve the development of cloned embryos. Round and smooth cells led to a high fusion rate. Cell types of donor and different individuals impacted on the development of cloned embryos.

A total of 360 newly-hatched AA broilers were randomly allotted into five dietary treatments with 6 replications of 12 broilers. The five experimential diets were consisted of corn-soybean meal basal diet without antibiotics as control treatment and the other four ones which supplemented the basal diet with 0.3% FOS, 0.1% Bacillus subtilis, 0.3% FOS + 0.1% Bacillus subtilis, and 22.5 mg/kg Aureomycin, respectively. The result indicated that dietary supplementation with FOS or Bacillus alone had selective effects on increasing caecal population of Lactobacillus and decreasing population of E. Coli and Salmonella. However, supplemental Aureomycin had non.selective effects on caecal microflora which inhibited all bacteria. Moreover, the combination of FOS and Bacillus had much better improvement on caecal micro.ecosystem for broiler cockerels. Fecal emission of ammonia and sulfureted hydrogen decreased by 38.38% (P<0.05) and 24.35% (P<0.05) resulted from addition of FOS and by 62.14% (P<0.05) and 28.49% (P<0.05) for combination of supplemental FOS and Bacillus, whereas no effect was significant (P>0.05) due to dietary supplementation with Bacillus or Aureomycin. Availability of crude ash increased by 18.94% (P<0.05), 17.36% (P<0.05) and 23.66% (P<0.05), calcium by 20.78% (P<0.05), 14.63% (P<0.05) and 21.31% (P<0.05), and Phosphorus by 6.60%(P>0.05), 12.32% (P<0.05) and 14.67% (P<0.05), respectively, resulted from dietary supplementation with FOS, Bacillus and combination of FOS and Bacillus.

The objective of this study is to determine the relationship between energy intake and abundance of apoE mRNA in liver of periparturient dairy cows. At 28 days before parturition, thirty multiparous periparturient Holstein cows were randomly divided into three groups fed with 100% energy diet (China Dairy Cow Raise Standard, 2000), 120% energy diet and 80% energy diet, respectively. After parturition, all the cows were offered criteria day provisions until 56 days postpartum. Relative expression of apoE mRNA in liver tissue was increased by degrees before parturition, peaked at 1 day after parturition and decreased gradually after parturition in the dairy cows fed with 80%, 100% and 120% energy diets. Relative expression of apoE mRNA in liver tissue of postpartum was higher than that of antepartum in three treatments(except for 56 days after parturition). Abundance of apoE mRNA in liver was significantly higher in the cows fed with 80% energy diet than in the cows fed with 100% and 120% energy diets from 14 days before parturition to 28 days after parturition（P<0.05或P<0.01） , and was significantly higher in the cows fed with 120% energy diet than in the cows fed with 100% energy diet from 28 days to 56 days after parturition(P<0.01). It is concluded that increasing energy intake in the dry cows could reduce the expression of apoE mRNA in liver, oppositely, reducing energy intake in the dry cows could increase the expression of apoE mRNA in liver of periparturient dairy cows.

Wheat gluten protein was added to the diet of goats to determine the changes of rumen movements with the rumen dynamic determination device. Compared with control period, the results of four goats in experiment period showed that, the average contract power of the goat’s rumen was significantly increased by 31.2%（P<0.05）,the average of rumen peristalsis continued time were prolonged significantly by 34.1%（P<0.05）in 15 minutes. As a contrast, rumen peristalsis frequency of the goats was decreased remarkably by 25.1%（P<0.05）. The determination device of rumen movement could sensitively observe change regulations of ruman movements in goats. At the same time, activities of opioid peptides from gluten protein degradated by rumen microbes were detected with the device of intestine of rabbits cultured in vitro. This provides theoretical base for the study on the control of rumen metabolism in ruminants.

In order to clone and study the cytokine and related genes of porcine, a cDNA library of porcine cytokine was constructed. The mononuclear cells were isolated from the peripheral blood and lymph nodes of healthy porcine and were stimulated by PHA+LPS at different time. The total RNA were extracted and the mRNA were isolated. Single-strand cDNA and double-strand cDNA were synthesized from mRNA, then ligated to directional EcoRI and HindⅢ linkers. cDNA were ligated into λScreen vector after size fractionating by gel filtration and packaged in vitro. The cloning efficiency was evaluated and the length of the cDNA fragment was assayed by PCR. Using the amplified library as template DNA, 2 pair primers were designed according to the sequence of the porcine IL2 and IL4, then the gene were amplified by PCR.The results demonstrated that a cDNA library of porcine cytokine has been constructed and the IL2 and IL4 gene were amplified successfully. The tilter of the cDNA library was 8×105 pfu/mL and the length of inserts was about 300~2 000 bp. It is helpful in the further study on screening novel cytokine and related genes.

Preparation and preliminary application of diagnostic gene chip of several avian diseases have been developed in this paper. Recombinant plasmids of target genes has been designed and cloned respectively. The target genes of NDV, IBV, AIV and IBDV were amplified by PCR with recombinant plasmids templates respectively and purified. Target genes were spotted on glass slides with microarray printing system, and the spotted slide was dried at room temperature, hydrated, lighted with UV and washed. The diagnostic gene chip for detecting NDV, IBV, AIV and IBDV were produced successfully. The results of qualification test showed that the diagnostic gene chip are reliable and well for detecting NDV,IBV, AIV and IBDV, and the sensitivity, specificity and repeatability of this new technology is well for detecting. 30 clinical samples were detected by gene chip technology and RT-PCR at the same time, the results illustrated that the detecting rate for NDV, IBV and IBDV are nearly same between the two ways, but the gene chip technology can detect different disease simultaneously.

A recombinant pseudorabies virus (PRV) (rPRV-HA) expressing the hemagglutinin (HA) gene from H3N2 subtype swine influenza virus (SIV) was evaluated in a mouse model. A total of 150 8-week-old female BALB/c mice were used in this study. The mice were each inoculated intranasally with 10^5.0 TCID₅₀ of rPRV-HA (rPRV-HA group, n=60) or attenuated vaccine Bartha-K61 (Bartha-K61 group, n=60), and another two unimmunized groups served as challenged group (n=20) or unchallenged group (n=10). Some mice in rPRV-HA and Bartha-K61 groups were sacrificed for antigen detection and virus isolation at different days post-immunization (DPI), and the others were each challenged with 10^5.0 TCID₅₀ of the same subtype SIV 28 DPI. Recombinant virus could be detected in the lung of rPRV-HA-immunized mice. Antibody responses to PRV were detected by indirect immunofluorescence assay, but not by seroneutralization test, in both rPRV-HA and Bartha-K61 groups. SIV-specific antibodies were detected by hemagglutination inhibition test only in rPRV-HA group 14 DPI. The rPRV-HA-immunized mice were protected from homologous SIV challenge, as indicated by limiting virus replication and pathological changes of the organs, and boosted antibody responses to SIV post-challenge.

The expression of neural and hematopoietic markers in placenta tissue and cultured human placenta derived adherent cells (hPDACs) and their correlation in embryonic development were investigated in this paper. The function of human placenta in hematopoiesis was demonstrated by immunocytological study in 7 and full-term months’placental slices. Positive reactions were observed in 7 month placenta using antibody against SCF, VEGF, BMP-4, FGF-1, CD34, CD133 and KDR. However, full-term months’placenta just reacted slightly with these antibodies. The staining for neural markers such as Nestin and MAP₂ showed positive reactions in 7 month and negative in full-term placenta. GFAP was showed negative in all two groups placenta’s dying. The isolated adherent cells of placenta were positive for AKP, vitamin, SCF, VEGF, BMP-4, FGF-1, CD133, Nestin and MAP₂ markers, while they were negative for CD34, GFAP and MBP. They showed neural differentiation potential under appropriate conditions. The data suggest that placenta can express hematopoietic related factors to support hematopoiesis, and placental-derived cells have neural differentiation potential.

Histology method and electron microscopy technique were used to investigate the changes of respiratory tract lymphoid tissues in pig during growth. The result showed that tonsil and pharynx were the first place of lymphoid tissue aggregation for respiratory tract entering the body. Lymphoid tissue in the tonsil existed at birth without any apparent lymph nodule aggregations. Lymphoid tissue began to increase at 20 days, with clear lymph nodule aggregations. The lymph nodules increased and densely packed beneath the stratified squamous epithelium of tonsils at 120 days. The well-developed lymph nodules were present with germinal center. A number of intraepithelial lymphocytes were located between the stratified squamous epithelium of tonsil. The bifurcation of trachea is the second place of lymphoid tissue aggregation for respiratory tract entering the body. The apparent lymphoid tissue in the adventitia of trachea bifurcation linked with tracheobronchial lymph nodule at birth, whereas they divided into two parts at 20 days: the packed lymphoid tissue in the adventitia of trachea bifurcation and tracheobronchial lymph nodule. The trachea bifurcation was rich with lymphoid tissue with increased intraepithelial lymphocytes at 120 days. More lymphocytes were located in the laminal propria of branches and bronchioles in the lung. The number of plasma cells increased and few intraepithelial lymphocytes existed between the columnar epithelium. Our results showed that the respiratory tract of pig was the effective induction sites and effective sites of mucosal immune. The local immunity in the respiratory tract can be enhanced by the nasal immunity in the piglet.

On the basis of New fertilitypromoting intrauterine infusion liquid (NFPL), 3 additive prescriptions were composed respectively by adding drugs of reducing fever, promoting blood circulation and replenishing Qi. Four prescriptions were used to treat rabbit experimental endometritis by intrauterine infusion and their curative effects were compared according to clinical pathological changes of symptom, uterine morphology, micro-and ultra-structure of endometrium after administration. The results showed that NFPL possessed significant curative effect. Clinical symptom of affected rabbits in NFPL group was abated, uterine perimeter and index decreased, endometrial inflammation extinguished and trauma recovered quickly. Prescription 2 was of better efficacy and could be used as an alternative of NFPL.

The anti-oxidative indexes in the serum and whey of the cow suffered from alcohol positive milk(APM) were detected. And the mechanism of APM was probed from the free radicals/ disorder in this test. Meanwhile, concentrations of microelement Zn,Cu,Mn,Fe in the serum and milk were detected and the relationship between concentrations of microelement and the free radicals/ metabolism was investigated. According to the result of experiment，excessive anti-oxidative reaction occurs and the total anti-oxidation capability goes down in the cow with APM. The system of anti-oxidation in the body can not eliminate the active oxygen and metabolites effectively，maybe the epithelial cells of mammary gland were mini-injured by the surplus of free radicals. It leads to the occurrence of alcohol positive milk.

Gene frequency, polymorphism information contents, number of effective alleles and locus heterozygosity were studied in 300 ducks of six populations (Shaoxing duck, Muscovy duck, Cherry duck, Peking duck (Z₁ and Z₄) and Aobaixing duck) using four microsatellite markers (AJ272577，AJ272578，AJ272579 and AJ272580). The result indicated that there are genetic polymorphisms at four microsatellite markers in six duck populations. Four microsatellite markers can be used for genetic diversity evaluation in duck populations. The genetic variability of AJ272578 is the highest, and AJ272579 is the lowest. The genetic variability of Peking duck (Z₁) is the highest, and Aobaixing duck is the lowest in six duck breeds.Based on Nei’s standard genetic distance, dendrograms were constructed using the unweighted pair group method with arithmetic mean (UPGMA)， Shaoxing duck and Muscovy duck were grouped together; Peking duck (Z₁), Aobaixing duck and Peking duck (Z₄) were clustered together, Muscovy duck was clustered another type. Microsatellite genotyping in ducks provided a useful tool for examining the genetic relationships among breeds (populations).

For exploring mechanism of drug-resistance of Trypanosoma evansi to antrycide, the RT-PCR and PCR were performed to clone the TbTA1 from the sensitive strain（TeS） and the resistant strain（TeR） under the different conditions of annealing temperature. The TbTA1 was obtained from the cDNA and DNA of the sensitive strain, but not obtained from the antrycide-resistant strain of T. evansi. The results suggested that the TbTA1 in the drug-resistant strain had some changes which result in antrycide-resistance and the TbTA1 of T. evansi, therefore, is involved in antrycide-resistance of T. evansi. On the other hand, the comparison of sequence of the TbTA1 between T. evansi and T. brucei revealed the 10 different nucleotides, G⁸⁸(T. e)-A(T. b), T¹⁴⁴(T. e)-C(T. b), C²²⁴(T. e)-T(T. b), C⁴⁷¹(T. e)-T(T. b), A⁴⁷²(T. e)-G(T. b), G⁵⁴⁹(T. e)-T(T. b), C^{1 008}(T. e)-T(T. b), A^{1 033}(T. e)-G(T. b), A^{1 293}(T. e)-G(T. b) and T^{1 384}(T. e)-C(T. b), resulting in Va^l30(T. e)-Ile(T. b), Ala⁷⁵(T. e)-Val(T. b), Ila¹⁵⁸(T. e)-Val(T. b), Thr³⁴⁵(T. e)-Ala(T. b) and Ser⁴⁶²(T. e)-Pro(T. b), these difference maybe due to different species.

In the present study, the ORF5M gene (the modified ORF5 gene) and ORF6 gene，the two major immunogenicity genes of PRRSV, were used as the target genes, and bovine herpesvirus 1(BHV-1 )VP22 possessing protein transduction property was chosed as the immunoadjuvant. Based on homologous recombination，a recombinant PRV (rPRV) TK^-/gE^-/VP22E⁺/VP22M⁺ coexpressing PRRSV E and M protein fused with BHV-1 VP22 protein was constructed by using PRV TK^-/gE^-/LacZ⁺ as parent strain. The rPRV was confirmed by PCR, Southern blot and Western blot. The results of TCID₅₀ tests showed that the insertion of the foreign genes had no influence on the propagation of rPRV in IBRS-2 or PK-15 cells. BALB/c mice were immunized with rPRV TK^-/gE^-/VP22E⁺/VP22M⁺, and compared with rPRV TK^-/gE^-/E⁺ expressing ORF5M gene and TK^-/gE^-/E⁺/M⁺ coexpressing ORF5M and ORF6 genes respectively. Neutralizing antibodies and lymphocyte proliferative responses were evaluated at 10 weeks after primary immunization. Among three rPRVs，TK^-/gE^-/VP22E⁺/VP22M⁺ had the best effect on enhancing PRRSV-specific humoral and cellular immune responses, and VP22 showed adjuvant efficiencies on rPRV vaccine to a certain degree. In conclusion, the study established a base for the research of bivalent genetic engineering vaccines against PRRSV and PRV.

According to the published nucleotide sequence of giardiavirus, six pairs of primers were designed and total nucleic acids from trophozoites of Giardia canis isolated from dogs in Changchun were used as template for RT.PCR. After RT.PCR, the products were linked into pMD18T vector, then cloned ,sequenced and analyzed. The genome sequence of Giardia canis virus was 6 276bp(DQ238861)，which revealed the presence of two large open reading frames that are separated by a -1 frameshift and share an overlap of 220 nt. The ORF1(367-3030) encodes a polypeptide of 887 amino acid residues. The ORF2(2808-5981) encodes a polypeptide of 1 056 amino acid residues,which contains all consensus RNA-dependent RNA polymerase sequence motifs. The ratio of G+C in the genome was 49.62%. The homology of Giardia canis virus to sequence of L13218(GenBank) was 94.62% in the nucleotide, and 93.50% in the amino acid levels. Compared with the sequences of AF525216(GenBank), the homology was 98.88% in the nucleotide, and 98.30% in the amino acid levels.

In this experiment, three recombinant expression vectors based on pVAX1 were successfully constructed. According to the sequences of spike (S) gene, membrane protein (M) gene and nucleoprotein (N) gene of CCV Insavc-1 strain, three pairs of specific primers were designed and used to amplify S, M and N genes of DXMV strain. The PCR products were cloned into pGEM-T. Then the recombinant pTS, pTM and pTN were used for sequencing. After sequenced, three immunogenicial genes were subcloned into pVAX1, respectively, which named pVAXS, pVAXM and pVAXN. The three constructed expression vectors were transfected into MDCK cell. Transcriptions of interesting genes were detected by RT-PCR. The expression of target proteins was detected by indirect ELISA. The results showed that target genes can be transcripted after 36 hours , and target proteins can be detected after 72 hours of transformation. Immunization in dogs indicated that the three expression vectors can efficiently induce the specific cellular and humoral immunoresponse, which laid a solid foundation for the further studies of new vaccines of CCV.