Avian coccidiosis is a major parasitic disease of economic importance in poultry industry and is controlled using prophylactic medication.The increasing incidence of drug resistance and the consumers’ increasing concern about the sanitation of poultry products prompt the development of vaccine against coccidiosis.Although being proved to have definitely effects against coccidial infection，live vaccine will be altered by molecular vaccine due to its possibility of rejuvenation.Hundreds of proteins involved in the process of invasion of the host cells have been described，seldom of them was proved to have a ideal effect against coccidiosis when the related genes were expressed in prokaryotic system.DNA immunization has been widely used in recent years as a novel vaccine delivery system to induce host high levels of cellular immune responses.

13 microsatellite loci were used to detect 60 horses of two horse breeds，namely mongolian and thoroughbred in this experiment. The PCR amplified products of microsatellite were detected by 8 percent nondenatured polyacry lamide gel electrophoresis and showing in silver staining protocol. Allele frequency，numbers of effective alleles（Ne），polymorphism information contents（PIC）and heterozygosity（H）for every microsatellite locus were calculated by softwares.The results were the greatest variation in UCDEQ440 and the lowest in TKY16 in 13 microsatellite loci；Mean numbers of Ne，PIＣ and H in thoroughbred horses were all higher a bit than in mongolian；Mean coefficient of gene differentiation（GST）of the whole breeds was 0.042 9 These results could provide references in the future that breeding new excellent breeds in our country.

The hybrid experiment has been proceeded between chicken and quail through the artificial insemination. The hybrids of layer(♂)—quail(♀), blackbone chicken(♂)—quail(♀) were gained successfully .The experiment indicated that the hybrids expressed strongly crossing sterility—early female embryo death，that is to say the differentiations of early cell stopped in female embryo. In order to study the genetic rule of cytoplasm in distance hybridization，the isolation of mitochondrial DNA from different viscera were largely founded in chicken,black—bone chicken,quail and their hybrids.The results showed that the intranuclear genome of hybrid and female parent appeared highly uniformity on mtDNA size, but different from their male. This paper analysized nine restriction endonuclease were applied to restriction fragment in mtDNA of chicken, blackbone chicken, quail and hybrids. The results indicated the hybrids have the same mtDNA haplotypes as their parent. This approved that maternal inheritance of mtDNA exist in chicken—quail interliner hybridization.

One hundred and eighty one-day-old Arbor Acre (AA) male broiler chicks were randomly allocated into three groups with six replicates each. From 8 days old, the chicks were supplemented with different levels of CoQ₁₀, i.e. 0, 20, and 40 mg/kg, respectively. During day 15 to day 21, all chicks were exposed to low ambient temperature(15~18 ℃) to induce ascites. The results showed that, under the condition of this trial, no significant effects were observed on broiler chicks’ growth performance, but the sensitivity of broiler chicks to ascites was significantly decreased by CoQ₁₀ supplementation. The ascites mortality of CoQ₁₀ supplemented groups was much lower than that of the control and EOF was significantly decreased by CoQ₁₀ supplementation. There were no significant changes observed on PCV between the control and the treatments. RVP, PAP and ±dp/dtmax were not influenced by the treatments except that PADP on day 36 was significantly decreased by 40 mg/kg CoQ₁₀ supplementation (P≤0.05). AHI was significantly reduced by 40 mg/kg CoQ₁₀ supplementation (P≤0.05). The beneficial effect of CoQ₁₀ may be attributed to its benefit for development of cardiac myocytes and decreasing EOF.

In vitro and in vivo studies were conducted to evaluate the efficacy of modified palygorskite (MP) to alleviate the toxic effects of aflatoxin (AF) B₁ in ducks.In vitro results indicated that MP was much more efficient to adsorb AFB₁ than palygorskite. In the in vivo study, four hundred and eighty Cherry Valley commercial ducks, 1 day old, were randomly allocated to four treatments for 42 days, which included: 1) Basal diet (control); 2) Basal diet supplemented with 0.25% MP; 3) Fifty percent of the corn in the basal diet was replaced by moldy corn and the concentrations of AFB₁ in prophase and anaphase are 50 and 55 μg/kg of diet, respectively; 4) The third diet plus 0.25% MP. The addition of MP to chick diets at a level of 0.25% had no significant effects on duck performance, AFB₁ contents in tissue and serum, organ weights, or serum biochemical index. In ducks fed the diet containing moldy corn alone, the performance was significantly decreased, the AFB₁ contents in breast muscle, liver, kidney or serum, the relative weights of the heart, liver, kidney, pancreas and the levels of serum glutamic-oxaloacetic transminase, glutamic-pyruvic transminase, amylase were significantly increased, and the concentrations of serum total protein, albumin, uric acid were significantly decreased. Ducks fed the combination moldy corn / MP diet had similar performance, AFB₁ contents in tissue and serum, organ weights, and serum biochemical index to those of control ducks. These results indicated that MP was effective in preventing the toxic effects of AF in ducks.

150 one-day-old broilers were divided into five groups randomly which were composed of three groups ad lib. and two zinc-sufficient pair-fed groups. The three groups ad lib. were fed respectively the different experimental diets which were supplemented with three levels of dietary zinc (0, 5, 80 mg/kg) and 3 000 IU/kg of dietary vitamin A in the zinc-deficient semi-purified diet which contained 4.67 mg/kg zinc and was mainly made of corn starch and low zinc soybean meal. In addition, two zinc-sufficient pair-fed groups with the zinc-deficient groups were restricted to the average consumed by zinc-deficient groups in order to further explore the effects of feed intake and growth restriction as a result of zinc deficiency in diet on vitamin A metabolism in broilers. The results indicated that zinc deficiency would result in a significant depression of feed intake and growth and serum zinc and vitamin A concentrations in broilers. A significant decrease of serum vitamin A concentration was a result of feed intake and growth restriction resulting from zinc deficiency when dietary zinc level was 9.67 mg/kg. However, significant decrease in serum vitamin A level was a result of not only zinc deficiency but feed intake restriction resulting from zinc deficiency when dietary zinc level was 4.67 mg/kg. Feed intake restriction would result in an apparent decrease in serum vitamin A concentrations in broilers, but it had no effects on serum zinc concentrations.

The effect of donor cell cycle, cell hypotonic treatment and recipient oocytes meiosis phase on development of bovine somatic cell nuclear transfer embryos was studied in the present experiment. Oocyte intraplasmic nuclear injection was used as embryo reconstruction method. The results indicated that: There was no significant effect of culturing bovine skin fibroblasts with 0.5% FBS media (attached quiescent cell) on the development of bovine nuclear transfer embryos as compared with nonquiescent cell (P>0.05).Treatment of donor cells with 0.1% sodium citric acid 20 s before nuclear injection was beneficial for cell membrane breakdown and nuclear dissociation, as a result , the nuclear transfer embryos development rate to 8/16-cell stage and blastocytes was significantly higher than control(P<0.01).It was found that donor cells suspended in 0.5% FBS TCM199 3 d (suspended quiescent cell) could make it easily to dissociate nuclear without hypotonic treatment, and development ability of the nuclear transfer embryos was also promoted slightly. The TⅡ oocyte (activation before nuclear injection)or MⅡoocyte (activation after injection) did not significantly affect the development of nuclear transfer embryos. Our findings suggest that donor nuclear dissociation efficiency was crucial to the sucsses of oocyte intraplasmic injection, suspension quiescence was a proper donor cell treatment method,both TⅡoocytes and MⅡ oocytes could be used as recipient.

Thirty-two purebred Lantang piglets of the same age derived from multiparous sow were slaughtered at the newborn, 3, 21 and 35 d of age respectively. Blood sample and tissues samples including liver and longissimus muscle were collected each time. Concentrations of IGF-1 in serum and in liver and muscle were determined by radio-immunoassay (RIA). Growth hormone receptor (GHR) specific binding activity in the membrane of liver cell and in muscle cell was determined by the method of radioligand binding assay(RBA). Real-time polymerase chain reaction (PCR) quantifications of IGF-1 mRNA and GHR mRNA from liver, muscle were carried out by DNA binding dye method using SYBR Green PCR Core Reagents Kit in Rotor-gene 2 000 Real-time Cycler. The results showed that: 1. Serum IGF-1 levels of newborn piglets were significantly higher than any other days observed (P<0.05).Concentrations of IGF-1 in longissimus muscle were higher than in liver per micrograms protein, longissimus muscle IGF-1 levels were higher at 3, 21and 35 d than that of newborn (P<0.05).2. Growth hormone receptor (GHR) specific binding activity in the membrane of liver cell was highest at newborn (P<0.05). Growth hormone receptor (GHR) specific binding activity in the membrane of liver cell was higher than in the membrane of longissimus muscle cell.3.IGF-1 and GHR mRNA levels in liver were significantly higher than longissimus muscle (P<0.05). Liver IGF-1 mRNA levels in newborn and 21 d were significantly higher than 3 d and 35 d(P<0.05). Liver GHR mRNA levels of newborn were significantly higher than any other days observed (P<0.05). Longissimus muscle IGF-1 and GHR mRNA levels of newborn were significantly higher than any other days observed (P<0.05).

This study has set up a network database on animal genetic resources in China depending on standardized resources data from bath “Chinese Animal Breed Records” and much collected animal genetic information anew, software and hardware circumstance and TBF no-structural full-text database. At the same time, we have issued our website http://www.cdad-is.org.cn/ by Internet. Inside the website, we supply many searching models such as full-text searching, field searching, combination searching, twice searching and multi-databases searching etc. Finally, it points out the future development objectives of the network database as follows: The first is how to get dynamic animal genetic information of China, the second is to realize transition of information serving from current content management to future knowledge management.

The retrovirus mediated transfer gene method was applied to integrate PrV Ea gD gene into the chromosome of PK-15，and the cell line designated as PK^D expressing gD was established.The gD expressed by PK^D distributed in the cell membrane and demonstrate the same immunoreactivity as the original gD.It was also noticed that the gD was in apparent polar distribution.PCR technique and indirect immunofluorescence assay were applied to detect PK^D,the results demonstrates that the transferred gD gene is genetically stable.In the same culture condition,the growth speed and the shape of PK^D did not show apparent difference compared with PK-15.By lipofeltin mediated method,PrV Ea genome was transferred into PK^D,a typical cytopathic effect was observed after 30 hours which proved PK^D has the endurance to lipofeltin.The detection results of TCID₅₀ demonstrated that although PK^D showed apparent suppression to PrV Ea strain,PrV Ea strain can be amplified in it normally.All this research results suggest that PK^D can be used to establish PrV Ea strain gD deleted mutant and thus provides an essential tool for it.

In order to amplify gag gene of Jaagsiekte sheep retrovirus Inner Mongolia strain, three pairs of primers were designed according to the GenBank sequence . The fragments of gag gene was obtained by polymerase chain reaction(PCR),then the genes were cloned into pMD-18 T vector and identified by PstI, EcoRI and SalI digestion. The nucleotide and amino acid sequences of NM strain gag gene were compared with the counterpart sequences of South Africa strain(NC-001494) and USA strain (AF105220). The nucleotide and amino acid homology of gag gene were 89.0%,90% and 86.3%,87%, respectively.

The virulence of twenty avian E.coli isolates to 1 day-old chicks was detected, and each isolates was examined for the presence of iss gene by using PCR techniques. Of the 13 virulent avian E.coli, iss were found in 12 isolates(92.31%), and 6 of 7 avirulent strains(14.29%) lack in iss, The results suggested that iss may take an important role in pathogenicity, which may be a marker of virulent-associated strains.

Pathogenesis were studied by inoculating the broiler chickens with reticuloendotheliosis virus(REV) and Marek’s disease virus (MDV) . After 13 days post-inoculation, broiler chickens began to die and the mortality reached 84% at the end of experiment. By gross examination, there were two different tumors of sporadic, large tumors and diffusible, small tumors that induced and mixed on the aspect of liver, spleen, kidney and heart. These samples were embedded with paraffin and cut into a series of segments that were stained with hematoxylin and eosin, and examined for existence of two different kinds of tumor cells by microscopic lesions. Some segments were reacted with monoclonal antibody of MDV BA4 and REV 11B118 respectively. The IFA examinations showed that both MDV and REV antibody could react with the same segment, which meant REV and MDV could induce tumors separately in chickens.14,21, 28 and 42 days after inoculated, both MDV and REV were isolated from three blood samples that were collected at random. The results showed that coinfection of MDV and REV could prolong the date of virus existence in the blood.

A gene encoding ferritin was isolated and cloned from B. microplus. The cDNA is composed of 642 nucleotides and encoding 172 amino-acid residues, the encoding region is between 123 bp to 639 bp, the deduced mass is 19.9ku and isoelectric point is 4.24.The deduced amino-acid sequence shows 93.60%, 88.37% and 83.72% similarities with the ferritin from tick Dermacentor variabilis, Ixoded ricinus, and Ornithodoros moubata respectively. The stem-loop structure of a putative iron responsive element is found in the 5-untranslated region of ferritin mRNA. The protein contains the conserved motifs for ferroxidase center. RT-PCR analysis suggests the ferritins are expressed in the eggs，larvas，fed female adult，engorgement female adult and male adult ticks of B. microplus.

The aims of this study were to analyze the genetic variation among strains of Toxoplasma gondii from different hosts and geographic origins in China, and to establish the foundation for further studies on molecular genetics and diagnostics of T.gondii. The internal transcribed spacer (ITS) and 5.8S of rDNA from ZS (human), SH (human), CN (pig), QH (sheep) and RH strains of T. gondii were amplified by PCR, cloned and sequenced. The results showed that ITS and 5.8S sequences of QH, ZS, SH, CN strains were identical, which were consistent with the sequences of the RH strain registered in GenBank; but the ITS2 sequence of the RH strain differed at two bases when compared with that of the other 4 strains. The results indicated that ITS sequences provide useful genetic markers for the identification and differentiation of T. gondii from other related protozoa, but the sequences are not suitable markers for studying the genetic variation within T. gondii.

Healthy chickens were fed with the transformed lactobacillus mixed in water from 0 to 11days of age, then challenged with the 5×10⁴ sporulated oocysts of E. tenella per bird. Pathologic changes were studied to evaluate the level of protection role of transgenic L. lactobacillu with SO7 gene. The results showed that the pathologic changes of transgenic group changed lightest and that the LaS107 transformed with SO7 gene has a good function to defense the E. tenella challenge. These researches make a good foundation for external genes expressed in lactobacillus and create a brand-new way for using genetic lactobacillus in human being and animals.

The influence of Toxoplasma gondii infection on the success of mice pregnancy was investigated in this paper. Immunohistochemistry was applied to locate immunocytes in the uterus, and enzyme-linked immunosorbent assay(ELISA) was conducted in order to determine the contents of tumor necrosis factor-α(TNF-α) and interleukin-10 in uterine lysates. All the mice were aborted when the Toxoplasma gondii were incubated intraperitoneously on day 0 or day 6 of pregnancy. A few CD8⁺T lymphocytes were scattered in the endometrium of infected animals, while none cells were seen in the control mice. Large number of F4/80⁺ macrophages were counted in the infected mice. A few were seen in the control group. TNF-α level was significantly higher(76.862±30.354 pg/mg protein) in the infected mice compared with the control animals(P<0.01),indicating a close correlation between pregnancy failure and Toxoplasma infection. IL-10 contents were 19.696±9.811 pg/mg protein in the supernatants of the mice infected with Toxoplasma on day 0, significantly lower than those of control group(48.856±16.518 pg/mg protein,P<0.05). No significant differences of IL-10 contents were observed between the infected mice and controls when infection was carried out on day 6 of pregnancy (P>0.05).

Four crossed-breed dairy goats were fed on Thermopsis Lanceolat R.br dry hay powder at dosage of 5 g/kg body weight,blood samples were collected on the 0th, 7th, 14th, 28th, 35th and 42nd experimental day　for examining routine blood indexes, serology enzymology and biochemical indexes. The clinical symptom of the dairy goats was observed every day and the goats were dissected to conduct pathological examination in the end of the experiment. Results showed that the toxic symptom of tested dairy goats occurred in the 30th experimental day with decreased weight, thin body, and coarse fur. In the 41st experimental day, the tested dairy goats began to move abnormally, and suddenly fell down to the ground when being driven away. In contrast to undosed period, the content of hemoglobin and the number of white cells of the tested dairy goats increased significantly (P<0.05) and activity of glutamic-oxalacetic transaminase (GTP) and alkaline phosphatase (APH) increased significantly (P<0.01). Urea nitrogen level in serum decreased significantly (P<0.01) from the 14th experimental day.Pathological changes mainly showed tissue organism was turgescent and bleeding. Histological examination mainly showed vacuolar degeneration in the tissue organism. It was the first time to observe collagenous fiber hyperplasy in liver by ultra structural examination.

The development of motor neurons in the ventral horn of spinal cord in prenatal goat were observed by light microscope and electron microscope. The results showed: (1)The form of motor neurons in the ventral horn of spinal cord underwent four developing phases: the undifferentiation neurons, the immature neurons, the youthful neurons and the ripe neurons; (2)With the development of the motor neurons the nucleus were enlarging，the heterochromatin was reducing and tending to distribute in the peripheral gradually. With the number of nucleolus decreasing, the central nucleolus occurred in eighth week, and it become to develop after that the karyotheca matured gradually; (3)The changes in the development of endomembrane system of the motor neurons accorded with that of motor neurons.

Effects of Bu Qi Huo Xue therapy on the treatment of Qi Xu Xue Yu syndrome in postpartum dairy cows were studied from the variation of hemorheology traits,oxygen free radical and nitric oxide free radical. The dairy cows were divided into three groups, 18 healthy postpartum cows, 9 postpartum dairy cows displaying Qi Xu Xue Yu syndrome, and 8 postpartum dairy cows displaying Qi Xu Xue Yu syndrome treated with herbs. The last group was fed with Gui-Qi-Yimu powder (Powder of chinese angelica, astragalus and motherwort) 5 times, at 0.8 g/kg according to the weight at 1, 2, 3, 5 and 7 d postpartum. Blood samples was taken to analyze the designed indexes of dairy cows at 1, 2, 4, 7 and 10 d postpartum.The results demonstrated that Gui-Qi-Yimu powder treatment significantly decreased ηb20s^-1（P<0.05）at 1 and 2 d, and ηp（P<0.01 or P<0.05）at 4, 7, 10 d; malondialdehyde contents at 1 , 2 and 7 d; as well as nitric oxide contents at 1 to 10 d of postpartum dairy cows with Qi Xu Xue Yu syndromes. Thus Bu Qi Huo Xue therapy can significantly improve the hemorheological status and maladjustment of oxygen free radical and nitric oxide free radical metaboly of postpartum dairy cows displaying Qi Xu Xue Yu syndrome.

A murine phage antibody library was constructed.First,the total RNA was extracted from fresh spleens of nonimmunized mice,then the cDNA library was achieved via reverse transcription PCR.Gene fragments encoding V_H and V_L were amplified and assembled into a single gene using a DNA linker encoding a polypeptide of 15 amino acid residues(Gly4Ser)3 through PCR.And finally the recombinant DNA fragments were cloned into the phagemid pCANTAB5E vector and introduced into E.coli TG1.The phagemid particles displaying functional ScFv were rescued by reinfection of helper phage M13K07,thus a murine antibody library was obtained.

This investigation was carried out to study the effect of glutamine supplementation on the jejunum morphological structure of broiler chickens. 288 1-day-old AA chicken were used in a completely randomized experiment with three treatments and three repetitions per treatment, Group Ⅱ and group Ⅲ, respectively, being supplemented with 0.4% and 0.8% glutamine and group Ⅰ, the control, not supplemented. Jejunum samples were taken at 1, 2, 3, 4 weeks of age for morphological studies under light microscope. The results showed, there were different effect of glutamine supplementation on villus height, villus width, crypt depth, muscle layer of mucous membrane thickness, muscle layer thickness and serous membrane layer thickness during all the experimental weeks of age. It was demonstrated that 0.4% and 0.8% glutamine supplementation significantly increased the villus height during all experimental weeks of age and the muscle layer of mucous membrane thickness at one and two weeks of age (P<0.05 or P<0.01), as well as increased the muscle layer thickness and the serous membrane layer thickness (P>0.05); However, glutamine decreased the villus width and the crypt depth (P>0.05).