In order to investigate the genotypic characteristics and biological characteristics of major capsid protein (VP1) of duck hepatitis A virus (DHAV) GX strain, the whole genome of GX strain was sequenced and compared with 3 serotypes reference strains of DHAV by the molecular biology software.The results showed that the full-length genome of DHAV GX strain was 7 800 bp, including 5'UTR (652 bp), 3'UTR (369 bp) and ORF (6 756 bp) encoding 2 251 aa and its coding products were 12 (VP0/VP3/VP1/2A1/2A2/2A3/2B/2C/3A/3B/3C/3D) at least.Sequence analysis showed that DHAV GX strain could be ranked DHAV 3 style.This strain shared the highest homology with DHAV-3 reference strains; Furthermore, GX strain and DHAV-3 FS strain were in the same cluster, might come from the same ancestor.Amino acid segments locating in 195 to 201 and 211 to 221 aa could be larvaceous dominant B-cell linear epitopes.It suggested that VP1 gene could be used as a candidate gene for the development of genetic engineering vaccine of DHAV.

In order to build more effective use of porcine circovirus type 2b (PCV2b) Cap protein on the diagnosis and control of diseases like porcine circovirus disease, the ORF2 gene which encoding porcine circovirus type 2b Cap protein was cloned into the prokaryotic expression vector pET-32a (+), constructing a recombinant plasmid pET-32a-ORF2, then the recombinant plasmid was transformed into E.coli BL21 and induced by IPTG to express recombinant protein, the recombinant protein was purified with nickel column.Then polyclonal antibody (PcAb) was prepared by immunization of rabbit with purified protein.ELISA, Western blotting, indirect immunofluorescence assay and the specificity experiments were used to detect the biological characteristics of the polyclonal antibody.The titer of polyclonal antibody was about 1:2¹⁶ detected by ELISA.Western blotting result confirmed that polyclonal antibody could react with PCV2b specially.Indirect immunofluorescence assay showed that polyclonal antibody was able to detect PCV2b in PK-15 cells, this result suggested that the polyclonal antibody had the ability to differential diagnosis of PCV2b.The specificity experiment showed that the polyclonal antibody only reacted with PCV2b, and did not react with PEDV, TGEV, PRRSV, PRoV, PRV and PPV.The polyclonal antibody against PCV2b prepared in this study provided a powerful tool for the study of etiological characteristics and clinic diagnosis of this disease.

In order to study Mongolian sheep Brachyury gene DNA sequence, main purpose of this article by using the method of PCR, DNA sequence of Mongolian sheep Brachyury gene cloning and the complete Brachyury gene sequences were obtained for the first time.The sequencing with NCBI BLAST function on the site, comparing the results with the published sequences of sheep Brachyury homology was as high as 99%.We compared these sequences with ten Brachyury genes of human, mouse and other animals in GenBank database.The results showed that the most sequences were identical.Mongolian sheep Brachyury shared a 96.30% amino acid homology with Bos taurus, the highest degree of homology indicated a close genetic relationship, and the Caudata had the lowest homology 58.45%.It could be further confirmed by phylogenetic analysis.Moreover, the results showed that Mongolian sheep Brachyury gene sequence of amino acids and other vertebrates Brachyury gene sequence of amino acids were highly conservative.The research results showed that Mongolian sheep Brachyury gene sequences and other vertebrates Brachyury gene sequences were highly conservative.

In order to study the differences of expression of FSHβ and FSHR genes respond to goose treated by counter-season production.The FSH levels in serum were detected.The pituitary, hypothalamus and ovarian was harvested from geese of experimental group and control group.The differences of FSHβ and FSHR genes expression in hypothalamus-pituitary-gonadal axis were detected by Real-time PCR.The results showed the FSH level of geese from experimental group was extremely significant higher than that from control group (P<0.01).FSHβ and FSHR genes were expressed in all the three tissues.The highest expression of FSHβ was in pituitary while the FSHR gene was in ovarian (P<0.01).In the different breeding cycle, the FSHβ mRNA expression in pituitary of experimental group was extremely significant higher than that in control group (P<0.01).The FSHR mRNA expression in ovarian was extremely significant higher than that in control group (P<0.01).Counter-season production treatment increased the expression of FSHβ gene in pituitary and FSHR gene in ovarian.The results indicated that FSH was highly associated with incubation maintenance.

In order to explore the function of osteopontin (OPN) regulated bone metabolism and the relationship between OPN and tumor, DNAMAN and Mega 5.02 softwares were used to analyze phylogenetic relationships of OPN protein, the OPN expression vector was prepared by RT-PCR and molecular clone; OPN was purified by SDS-PAGE gel slices, renatured by urea concentration gradient, and immunized mice to prepare the polyclonal antibody; The antibody titer and specificity were detected by ELISA and Western blotting, respectively.The homology comparison result of OPN sequence showed that it existed short repeat sequenc Arg-Gly-Asp (RGD) in different animals with evolution levels.Phylogenetic tree of OPN sequence showed that OPN had obvious evolution trend among animals.RNA was extracted, OPN recombinant vector had been digested and sequenced to confirm the correct construction, and strip lengths of OPN expression and purification were agreed with the prediction.The OPN antiserum titer was 1:1 600 detected by ELISA, and Western blotting proved that the antiserum had good specificity.We had successfully analyzed genetic evolution relationship of OPN sequence, gotten OPN expression vector, purified and renatured OPN, and prepared and identified the polyclonal antibody against OPN.

Mitofusin-2 (MFN2), an outer transmembrane protein on the mitochondrion, which was a signaling and therapeutic target molecule, played a critical role in cell proliferation, differentiation, apoptosis and many diseases.For further researching the molecular mechanism of MFN2 gene in inflammatory disease, the MFN2 gene from Boer goat was cloned by RT-PCR, and its sequence and coding protein structure were analyzed.The results showed that the full length of MFN2 gene was 2 441 bp and its coding protein was composed of 705 amino acids.It was low homology between species with conservative protein structure, Fzo-mitofusin structural domain and DLP-2 structural domain, respectively.The MFN2 had a potential transmembrane domain, and was consist of 576 to 595 amino acid without signal peptide.It was the first time to clone MFN2 gene from Boer goat, which provided basic data for exploring the molecular mechanism of the relationship between MFN2 gene from Boer goat and inflammation.

Enterotoxic Escherichia coli (ETEC) can cause acute diarrhea in human and animals, and its key virulent factor is heat-stable enterotoxin (ST).ST is a peptide with small molecular weight and great toxicity but without immunogenicity, so how to keep its immunogenicity and reduce its toxicity has become a hot research topic for preparing ST toxoid vaccine.In this study, rabbit serum albumin (RSA) was purified by rivanol method and activated in 0.2% glutaraldehyde solution before RSA was conjugated to synthetic methanol-soluble STa, SDS-PAGE result showed that an approximate 108.5 ku complete antigen was obtained.New Zealand White rabbits were immunized with the conjugate four times, the serum was then collected and used to purify immunoglobulin by immunoprecipitation with Protein A/G Plus-Agarose.Dot-ELISA and Western blotting result showed that the titer of the antiserum both reached 1:400.These results indicated that anti-STa polyclonal antibody was successfully prepared, which provided the way for screening of ST structure analogues.

The PCR technology was used to amplify the E5 gene according to the genome sequence of bovine papillomavirus type 13 (BPV-13) Hainan strain with the GenBank accession number KM258443.2.Then it was ligated into pEGFP-N1 vector.The recombinant plasmid pEGFP-E5-N1 was transfected into HEK293 cells with liposomes by transient transfection.The expression of fusion protein was identified by fluorescence microscope, flow cytometry and Western blotting.The results showed that the E5 gene was 135 bp with the 100% homology with the sequence deposited in GenBank.Bright green fluorescence could be seen in HEK293 cells transfected by pEGFP-E5-N1.The expressed fusion protein was about 32 ku.This experiment laid the foundation for further study of the functions of E5 gene in eukaryotic cells.

The study was aimed to establish a specific and sensitive TaqMan Real-time PCR assay for detection of Arcanobacterium pyogenes (A.pyogenes).Based on the conservative sequence of pyolysin (PLO) gene of A.pyogenes published in GenBank, specific primers and TaqMan probes were designed. The TaqMan Real-time PCR assay was established, and the specificity and sensitivity were tested.The specificity test results showed that only A.pyogenes exhibited typical curves.The detection sensitivity of this assay was 77.6 fg genomic DNA per 20 μL reaction, and 63 CFU/mL for pure cultures.16 out of 23 clinical samples were positive detected by the TaqMan Real-time PCR assay, which were consistent with API Coryne identification.The TaqMan Real-time PCR assay developed in this study was specific and sensitive for detection of A.pyogenes, and it could be used for identification and epidemiological investigation of A.pyogenes.

In order to develop a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of Torque teno sus virus type 2 (TTSuV2), two pairs of primers were designed according to the untranslated regions and the part of open reading frame 1 of TTSuV2.The LAMP system was optimized by adjusting the concentrations of some components and reaction conditions.The optimized amplification conditions of LAMP assay was at 64 ℃ for 90 min.The results showed the LAMP assay was specific for TTSuV2 detection, which could achieve a detection limit of 100 copies/μL viral nucleic acid, and no cross-reaction with TTSuV1, PCV2, CSFV, PRRSV and PBoV.In conclusion, this assay was a rapid, specific and sensitive detection technique which could provide a assistance for the rapid detection of TTSuV2.

Rotavirus is a major cause of acute diarrhea in both many kinds of young animals and children under 5 years old.Rotavirus NSP1, a 55 ku RNA binding protein, is the product of gene 5, which can subvert innate immune responses and be one of virulent determinant factors.According to the sequence in GenBank, specific primers targeting to NSP1 gene were designed and the gene was amplified by RT-PCR, following by being cloned into the pET-28a(+) vector.It showed that the full length of NSP1 gene was 1 473 bp, encoding 491 amino acids.The NSP1 shared the highest identity with WC3 strain.The recombinant protein was induced in E.coli Rosetta(DE3) by IPTG and was analyzed by SDS-PAGE and Western blotting.The results revealed that NSP1 recombinant protein existed in the form of inclusion body with the molecular weight of 55 ku.The purified recombinant protein could be recognized by His-tag antibody.This study laid the foundation for further research on the relationship between the intracytoplasmic location of NSP1 protein and its activity.

With purified pseudorabies virus (PRV) to immune BALB/c mice, spleen cells from imunized mice were fused with SP2/0 myeloma cells by application of lymphocyte hybridoma technique, followed by double antibody sandwich ELISA screening, four times cloning by limiting dilution method to obtain two stable secreting anti-PRV monoclonal antibody hybridoma cell lines, 2C6E6 and 3D5F1.After identification, these two monoclonal antibodies belonged to IgG1 and IgG2a subtypes, respectively, the average number of chromosomes of hybridoma cells was 84, ELISA titers of these two hybridoma cell culture supernatants and mouse ascites monoclonal antibodies were 1:512 and 1:1 638 400, 1:1 024 and 1:819 200.These two monoclonal antibodies showed no cross-reaction with classical swine fever virus, porcine reproductive and respiratory syhdrome virus and porcine parvovirus, which indicated that they had high specificity.The hydridoma cells could passage for 30 generations which could still secrete monoclonal antibody against PRV, which indicated that these two monoclonal antibodies had good stability.The results in the assay laid the foundation for establishing rapid diagnostic methods of PRV.

Myocyte enhancer factor 2B (MEF2B) gene belonged to myocyte enhancer factor 2 (MEF2) gene family.They all widely expressed in muscle and nerve tissues of human and animals.It played an important role in the growth of muscle, development and differentiation of nerve system and liver fibrosis.This review mainly focused on the structural characteristics, tissue distributions, biological functions and research progress of MEF2B gene in human and animals.

This test mainly studied the effects of supplemental different levels of oil on the performance, plasma antioxidant capacity and other metabolic index of Yanqi horse.9 Yanqi horses were randomly divided into three groups according to performance, body weight, body measurement, age.The control group was fed with basal diet.Horses in the groups Ⅰ and Ⅱ were fed with diet supplemented with 2.5% and 5% of oil.All horses were participated in 20 km racing, on the 20th day.Temperature, heart rate, respiratory rate of all horses were measured, 30 min after the racing.Simultaneously, blood samples were collected from the jugular vein.The results showed that the heart rate value of groups Ⅰ and Ⅱ were highly significant lower than control group (P<0.01), 0 min after the racing.The group Ⅱ was highly significantly lower than control group (P<0.01), 5 min after the racing.The plasma GSH-Px and T-SOD activity of groups Ⅰ and Ⅱ compared with the control group respectively increased by 87.60%(P<0.05), 15.87%(P>0.05) and 98.38%(P<0.05), 38.25%(P<0.05);The plasma MDA and LA concentration of groups Ⅰ and Ⅱ compared with the control group respectively decreased by 53.44% (P<0.05), 24.08%(P<0.05) and 51.42% (P<0.05), 25.08%(P<0.05).In conclusion, the performance and antioxidant capacity of Yanqi horse could be improved by the basal diet added different levels of oil.The physiological function of Yanqi horse could be fast recovery after the racing.Compared, added 5% oil feed supplement was especially effective.

This study was conducted to investigate the effects of N-carbamylglutamate (NCG) and arginine (Arg) on growth performance, serum biochemical indexes and serum free amino acid content of Jinghai Yellow broiler chicks.A total of 600 1-day-old chicks with similar weight were used and randomly assigned into 5 groups with 6 replicates per group and 20 chicks per replicate.The control group was fed with basal diet, groups Ⅰ to Ⅳ were fed with the basal diets supplemented with 0.05%, 0.10%, 0.15% NCG and 0.10% Arg, respectively.The trial lasted for 28 days.The results showed that compared with control group, the supplementation of 0.05%, 0.10%, 0.15% NCG and 0.10% Arg increased ADG, the supplementation of 0.10% NCG significantly decreased F/G (P<0.05).The supplementation of 0.10% NCG significantly decreased the contents of AMM, UA and LDL in serum compared with other groups (P<0.05), the supplementation of 0.15% NCG significantly increased the contents of NO, NOS and ALP in serum compared with control group (P<0.05).Compared with control group, the supplementation of 0.10%, 0.15% NCG and 0.10% Arg significantly increased the content of Arg in serum (P<0.05).These results indicated that supplementation with NCG could increase the contents of Arg and Pro in serum, improve blood parameters and growth performance of broiler chicks, and the appropriate supplemental level of NGG was 0.10%.

In this paper, thirty Yili mares aged 11 to 12 years with weighing 410 kg ± 30 kg, parity of 4 to 5 in third lactation month were selected, and were randomly divided into six groups, each group with five repeats.Two factors quadratic regression orthogonal rotational combination design was adopted to evaluate the effects of feeding different dietary CP and DE to mares on growth development and blood biochemical parameters of foals.Daily DE and CP feeding levels of the 6 groups were 92 MJ/d and 1.20 kg/d, 125 MJ/d and 1.20 kg/d, 92 MJ/d and 1.55 kg/d, 107 MJ/d and 1.35 kg/d, 125 MJ/d and 1.44 kg/d, 115 MJ/d and 1.55 kg/d, respectively.The results showed that body weight, body height, chest circumference and cannon circumference of foals of four and five months were not significant affected by maternal protein and energy intake (P>0.05).As compared with ration contained high DE (125 MJ/d) and low CP (1.20 kg/d), feeding with ration contained moderate DE (107 MJ/d) and CP (1.35 kg/d) improved body length of foals of four and five months significantly (P<0.05).From the blood biochemical parameters, such as total protein, albumin, triglyceride, high-density lipoprotein cholesterol, low density lipoprotein cholesterol, urea nitrogen, creatinine, alanine aminotransferase, there were no significant differences among all groups (P>0.05).Mares fed with ration contained moderate DE (107 MJ/d) and CP (1.35 kg/d) obviously improved the aspartate aminotransferase and uric acid level.In summary, Yili mares during lactation late stage fed with 1.35 kg/d CP and 107 MJ/d DE daily could significantly improve body length development of foals, and foals nitrogen metabolism was altered by maternal dietary nutrition through increasing blood aspartate aminotransferase activity.

This experiment was conducted to investigate the effect of fermented peanut shell on production performance, carcass traits, meat quality and economic benefits of fattening pigs.100 fattening pigs (Duroc×Landrace×Yorkshire) with an average weight of 50 kg were randomly divided into 5 groups with 4 replicates per group and 5 pigs per replicate.The control group was fed with corn-concentrates-based feed, groups 1 to 4 were fed with the basal diets supplemented with 10% fermented peanut shell, 20% fermented peanut shell, 30% fermented peanut shell and 15% fermentation of corn stalk, respectively.The trial lasted for 66 days.The results showed that ADFI in control group, group 2 and group 3 were significantly higher than that in group 1 and group 4 (P<0.05), ADG in control group and group 3 were significantly higher than that in group 1 and group 4 (P<0.05), there was no significant difference in F/G among all groups (P>0.05).Diarrhea frequencies in groups 1 to 4 were extremely significantly lower than that in control group (P<0.01).There were no significant differences in carcass quality, slaughter rate, backfat thickness and eye muscle area among all groups (P>0.05), lean meat percentage in group 2 was significantly lower than that in control group and group 1 (P<0.05).There were no significant differences in marbling, meat color, tenderness, intramuscular fat, water loss and cooked meat percentage among all groups (P>0.05), water losses in groups 1 to 4 were lower than that in control group, but the differences were not significant (P>0.05).According to the economic benefit analysis showed that in pig from 50 to 90 kg, average profit of each pig in control group was 47.60 yuan, average profits of each pig in groups 1 to 4 were respectively 56.40, 60.40, 78.80 and 58.80 yuan, while group 3 was optimal.Based on the analysis of the above result, the fermented peanut shell had no bad effect on growth performance, carcass traits and meat quality of pigs, and it could effectively reduce the diarrhea frequency of pigs, improve the system of hydraulic of pork, when the fermented peanut shells were appropriatly applied in pigs feed, which could effectively reduce the breeding cost, and improve the economic benefit.

In order to study the effect of lactic acid bacterium on liver performance, fat deposition, slaughter performance, nutrient apparent digestibility and serum biochemical indices of Landes geese, a total of 108 12-week-age healthy Landes geese were selected and randomly divided into 3 groups with 6 replicates per group and 6 geese per replicate.The geese in control group were fed with the diets without lactic acid bacterium during the whole experiment, and the geese in groups A and B were fed with the diets supplemented with 2 000 and 3 000 g/t lactic acid bacterium during 1 to 5 d, respectively;With 3 000 and 5 000 g/t during 6 to 15 d, respectively;And with no lactic acid bacterium during 16 to 25 d.The results showed that compared with the control group, supplementation of lactic acid bacterium could prolong the mature period of liver (P<0.05) and decrease the elimination rate of Landes geese.Compared with the control group, supplementation of lactic acid bacterium could decrease the rate of fat on liver, the average weight of liver, liver weight/weight of bowel and abdominal fat and liver weight/weight gain in group B were significantly decreased (P<0.05).There were no significant differences in slaughter performance among all groups (P>0.05).Compared with the control group, the apparent digestibility of crude fat in group B was significantly increased (P<0.05), the apparent digestibility of calcium and phosphorus in groups A and B were significantly increased (P<0.05).Compared with the control group, the serum content of triglyceride, cholesterol and very low density lipoprotein in groups A and B were significantly decreased (P<0.05).In conclusion, diets with lactic acid bacterium were good to geese, but considering its prevention of fat deposition in liver, lactic acid bacterium was suggested not to be used in force-feeding process.

This study was conducted to investigate the effect of nutrients on laying rate, egg quality, blood biochemical indices and certain mineral elements of laying hens under free range system.100 laying hens with similar age and weight were respectively selected from each farm, Hantai, Heishanzhai, Tailing in Changping and Doushi, Liyuan in Shunyi poultry farm, and randomly allotted to 2 groups with 5 replicates per group and 10 hens per replicate.The control group was fed with corn as the main diet and test group was fed with corn as the main diet supplemented with appropriate of trace elements, vitamins and other nutrients.The trial lasted for 52 days.The results showed that the laying rate of test group was significantly higher than that of control group in different farm (P<0.05).The egg weight of test group was significantly higher than that of control group in Hantai, Doushi, Tailing and Liyuan (P<0.05).The protein height of control group was significantly higher than that of test group in Liyuan (P<0.05).The yolk color of test group was significantly higher than that of control group in Doushi and Liyuan (P<0.05).There were no significant differences in the egg index, shell thickness and average shell strength among all groups (P>0.05).The plasma contents of TP and TG of test group was significantly higher than that of control group in Heishanzhai (P<0.05).The content of Ca in egg of test group was significantly higher than that of control group in Hantai and Doushi (P<0.05), the contents of Cu and Mg of egg in Doushi and the contents of Mn and Fe of egg in Hantai of test group were significantly higher than that of control group (P<0.05), the content of Mg in egg of test group was significantly higher than that of control group in Heishanzhai (P<0.05).In conclusion, adding appropriate level of nutrients could improve the laying rate and the contents of Ca, Zn, Mg and other minerals in egg, increase the plasma contents of TP, TG and CHO, and significantly improve the nutritional status of laying hens.

In order to research the dynamic changes of nutrient intake and serum biochemical indexes of barn feeding ewes in different pregnant stages, and provide evidence for reasonable diet formulation and health state analysis of barn feeding goats, each 10 healthy female goats in non-pregnancy, early pregnancy and late pregnancy were randomly selected in this experiment, whose feed intake were determined;6 ill parturient ewes and each 8 healthy female goats in non-pregnancy, 1 month pregnancy, 2 months pregnancy, 3 months pregnancy, 4 months pregnancy and labor were randomly selected in this experiment, whose 7 serum biochemical indexes were determined.The results showed as follows:① In comparison to NRC(2007) goat nutritional requirements, DM, ME and CP intakes were more than the requirements in non-pregnant and early pregnant ewes, while DM, ME and CP intakes were less than the requirements in late pregnant ewes;Ca and P intakes were much more than the requirements in various stages of ewes.② From non-pregnancy, pregnancy to labor, healthy ewes' GLU content decreased (3.44 to 2.57 mmol/L) and KB content increased (14.93 to 23.55 μg/mL), while ill parturient ewes'GLU content was significantly decreased to 0.85 mmol/L(P<0.05) and KB content was significantly increased to 42.98 μg/mL (P<0.05);Healthy ewes' TP content decreased (70.55 to 59.94 g/L) and ALB content had little change (33.55 to 32.61 g/L), while ill parturient ewes' TP and ALB content both further decreased (56.79 and 29.72 g/L, respectively);Healthy ewes' Ca and P content had little change (2.21 to 2.19, 2.62 to 2.25 mmol/L, respectively), while ill parturient ewes' Ca (1.80 mmol/L, P<0.05) and P (2.17 mmol/L) content both decreased;Healthy ewes'AKP content decreased (258.57 to 104.35 U/L), but ill parturient ewes'AKP content increased (180.53 U/L).All the analysis showed that the inadequate energy intake of ewes in late pregnancy was the root cause of the decreased GLU and other serum biochemical indexes contents and the increased KB contents and the ill parturient ewes was diagnosed as pregnancy toxemia.Therefore, the ewes'diet formulation should be adjusted, different stages of ewes should be separated to feed, the energy intake of ewes in late pregnancy should be increased.

The objective of the present study was to investigate the influence of castration on daily gain and body size of Simmental cattle.50 healthy Simmental cattle (16 month) were used in a paired design of single factor by body weight.The results showed that 17 to 21 months of age, body weight and the average daily gain of castrated groups were lower than uncastrated group, respectively, there were no significant difference (P>0.05).17 to 21 months of age, body height, body length, chest circumference and hip width's length and month growth of the castrated group and uncastrated group, were all no significant difference (P>0.05).The first months after castration (17 month of age), body height, hip width and ischial-end width's month growth of castrated group were extremely significantly less than uncastrated group (P<0.01).17 to 19 months of age, ischial-end width of castrated group was significantly less than uncastrated group (P<0.05).The whole period of average month increase of abdominal perimeter, castrated group was more than in uncastrated group (P<0.01).The results suggested that the whole period of average daily gain of castrated Simmental cattle was less than uncastrated.The growth curve of castrated Simmental exponential was exponential curve, and uncastrated Simmental was "S".Inhibated the growth of body height, hip width and ischial-end width shortly after castration.The castrated would improve Simmental abdominal perimeter growth.Simmental body size at the age of 1.5 was close to the adult body size.

This test was designed to study the expression of POTs (PepT1, PepT2, PHT1 and PHT2) mRNA in calves'tissues.Tissue-specific expression of the mRNA corresponding to peptide transporter protein in four 3-month-old Chinese Holstein calves was examined by relative quantitative RT-PCR analysis.The results showed that the expression of PepT1 mRNA in the rumen was extremely significantly higher than that in the heart and muscle (P<0.01);The expression of PepT2 mRNA in the liver and kidney was extremely significantly higher than that in the heart, spleen, thymus, muscle, rumen, reticulum, omasum and abomasum (P<0.01);The expression of PHT1 mRNA in the lung and spleen was extremely significantly higher than that in the heart and muscle (P<0.01);The expression of PHT2 mRNA in the lung and thymus was significantly higher than that in the heart, liver, kidney, spleen, muscle, rumen, reticulum, omasum and abomasum (P<0.05).The results indicated that PepT1, PepT2, PHT1 and PHT2 mRNA were respectively abundantly expressed in rumen, kidney and liver, spleen and lung, lung and thymus.

Effects of Pseudostellaria polysaccharides on the contents of intestinal mucosal SIgA, IL-2 and IL-6 in mice were reported in this paper.36 Kunming male mice with 18 to 22 g were randomly divided into six groups, blank control group, Pseudostellaria polysaccharides control group, cyclophosphamide (CY) model group, and Pseudostellaria polysaccharides low, middle, high dose groups.Each mouse in Pseudostellaria polysaccharides control group and Pseudostellaria polysaccharides low, middle, high dose groups was respectively treated with polysaccharides at the dose of 400, 100, 200 and 400 mg/kg body weight by intragastrical treatment for 19 continuous days, CY was given to each mouse in CY model group and Pseudostellaria polysaccharides low, middle, high dose groups by intraperitoneal injection with 100 mg/kg body weight in the 20th day, duodenum and ileum samples were collected from mice after 24 h, and SIgA, IL-2 and IL-6 contents were determined.The results showed that SIgA contents in duodenum were significantly increased (P<0.05) in Pseudostellaria polysaccharides middle and high dose groups compared to CY model group, SIgA contents in ileum were extremely significantly increased (P<0.01) in Pseudostellaria polysaccharides high dose group compared to CY model group.IL-2 contents in duodenum and ileum were significantly increased (P<0.05), IL-6 contents in duodenum were extremely significantly increased (P<0.01), IL-6 contents in ileum were significantly increased (P<0.05) in Pseudostellaria polysaccharides high dose group compared to CY model group.It was concluded that Pseudostellaria polysaccharide could antagonize intestinal mucosal injury caused by CY.

The assay was aimed to explore the feeding and the best method to establish allergic rhinitis (AR) model in Hartley guinea pigs.Using the single factor experiment, the assay studied the effects of animal species, feeding way, ovalbumin (OVA) sensitization dose, time for AR model, and observed allergic reaction symptoms and nasal mucosal pathological section of Hartley guinea pig.The results showed that Hartley guinea pig was the best experimental animal for the AR experiment of nasal drops;Feed was ordinary mice feed + lettuce leaves or leaf lettuce;OVA as the sensitizer, concentration of basic sensitization was 0.1 to 0.3 mg/mL OVA, the next day intraperitoneal injection 1 time, for 7 times;Concentration of stimulated sensitization was 2% OVA, 10 to 30 μL each nostril, 1 time a day, after 6 to 9 d, compared with the blank group, the Hartley guinea pig model group had significant allergic symptoms and nasal mucosa lesions.This modeling method was simple and economic, good reproducibility, and could be used as the best method for AR model establishment of nasal drops.

The research was aimed to study the stability of avilamycin with different formulations.The relative contents of avilamycin and two main components were determined by high performance liquid chromatography (HPLC).Temperature and light tests were used to study the change rule of the relative content of avilamycin and two main components, so that the stability of avilamycin could be evaluated.The results showed that the formulation of avilamycin premix and avilamycin active pharmaceutical ingredients (API) were sensitive when stored in the conditions of different temperature and light conditions.And avilamycin API and avilamycin A component were more stable than avilamycin premix and avilamycin B component, respectively.So avilamycin should be stored at blow 20 ℃ and no light sealing, which could effectively improve the stability and quality of the product.

In order to evaluate the toxicity of Wu Jin granules for the guidance of clinical treatment, the acute and sub-chronic toxicity were assessed in KM mice and Wistar rats, respectively.The results of acute toxicity test suggested that the LD₅₀ of Wu Jin granules was >40 g/(kg·BW), maximal tolerance dose was 160 g/(kg·BW), equivalent to 80 times of clinical dosage.In sub-chronic toxicity test, the growth and general behavior of the animals appeared normal.Compared with the control group, weight gain and feed consumption of Wistar rats in the treatment groups had no significant difference (P>0.05).In high dose group, serum bilirubin (T-BIL), total protein (TP), albumin (ALB), urea nitrogen (BUN) and creatinine (CREA) of female Wistar rats, whereas CREA levels and liver index of male Wistar rats were significant difference (P<0.05), and the differences in other indexes were not significant (P>0.05); Hematological indexes, biochemical indexes of blood and organ index of Wistar rats in low dose group and medium dose group were not significantly different compared to the control group (P>0.05).Pathological examination results showed that mild granular degeneration existed in liver of Wistar rats in high dose group, whereas appeared clear and normal organizational structure of liver in Wistar rats in other groups.Wu Jin granules could inhibit intake of free bilirubin and hepatic synthesis protein in high dosage, whereas this results were not observed in other groups.Thus, the Wu Jin granules were safe in clinical treatment.

The study was aimed to research the growth inhibitory effects of two new bioactive peptides Temporin-Lb and Catesbeianin-1a from Rana catesbeiana on human lung cancer NCI-H446 cells, breast cancer MCF-7 cells and mice leukemia K562 cells, and provide the basis for selecting the new peptide antitumor drugs.The second structures of two bioactive peptides were tested by circular dichroism spectrum (CD), and the effects of the two new bioactive peptides on human lung cancer NCI-H446 cells, breast cancer MCF-7 cells and mice leukemia K562 cells were examined with MTS cytotoxicity assay.The circular dichroism spectrum results showed the secondary structure of Temporin-Lb was PPⅡ, and the secondary structure of Catesbeianin-1a was random coil.Using MTS cytotoxicity assay, it was found that given Catesbeianin-1a, the sharp of the three cancer cells above had little changed after culturing for 24 h, and the three cancer cells promoted normal.Given Temporin-Lb, the cell morphology of three cancer cells had changed after culturing for 24 h, the growth of the three cancer cells above had been inhibited, especially the bioactive peptide Temporin-Lb had a sharp antitumor effect to mice leukemia K562 cells between the concentration of 4 and 40 μmol/L.Bioactive peptide Catesbeianin-1a had no obvious effect on proliferation of the three cancer cells above.Bioactive peptide Temporin-Lb had a certain inhibitory effect to tumor cells.

The assay was aimed to establish high fat diet induced metabolic syndrome (MS) model in male SD rat of nutrition.Four weeks old male SD rats were randomly divided into experimental group (n=24) and control group (n=6).The control group was fed with normal diet and the experimental group was fed with high fat diet which was composed of normal diet, egg yolk and peanuts.Two weeks later, the obesity resistant rats were excluded from the experimental group.Weight, systolic blood pressure and Lee's index were observed after thirty-two weeks.After twelve hours fasted, made the OGTT.Then all rats were sacrificed and tested for total cholesterol (TC) and serum triglyceride (TG).The experimental group weighted (794.0±63.5)g and the control group weighted (571.8±61.9)g, and the weights of rats of experimental group were 38.9% higher than that of control group.Lee's index of the experimental group was 343.0±8.7, while the control group was 319.2±7.1 (P<0.01).The levels of serum TC and TG of experimental group were extremely significantly higher than that of control group (P<0.01).Systolic blood pressure of the experimental group was 140.0±15.4, while the control group was 117.9±11.4, differences comparison of systolic blood pressure between experimental group and control group were significant (P<0.05).Insulin resistance of the experimental group was significantly (P<0.05).This study showed that four weeks old male SD rats were fed with high fat diet which was composed of normal diet, egg yolk and peanuts for thirty-two weeks could successfully and stably established metabolic syndrome model.

In order to research the genetic diversity and phylogenies of Wuzhishan pig, 56 complete mtDNA D-Loop were analyzed.55 sequences were amplified by polymerase chain reaction (PCR) and 1 sequence was downloaded from GenBank.The results showed that repetitive sequence "TTATAAAACAC" appeared in the conservative regions of 21 mtDNA D-Loop sequences.13 haplotypes and 14 variable sites were observed in 56 sequences.Haplotype diversity and nucleotide diversity (Hd =0.838 and Pi=0.00334) indicated that Wuzhishan pig had high genetic diversity.Two branches in the phylogenetic tree and Network analysis diagram indicated that there were two maternal origins in Wuzhishan pig.The phylogenetic tree of Wuzhishan pig and other 23 species showed that Wuzhishan pig had close genetic relationship with the pig breeds in the south region of the Yangtze river.

To understand the relationship between single nucleotide polymorphism sites (SNPs) of myocyte enhancer factor 2A (MEF2A) gene and slaughter traits in Sansui duck, a total of 60 individuals of Sansui ducks were selected to investigate in this study, direct sequencing of PCR and PCR-SSCP methods were used on single nucleotide polymorphisms of MEF2A gene, and genetic effects of its on slaughter traits were analyzed.The results showed that two SNPs which include g.47915G>A and g.47918G>A of exon 11 were found in MEF2A gene, and the G/A mutation in the g.47915G>A SNP resulted in the change of codon from GAA to AAA, and the coding amino acid from Glu to Lys, and the G/A mutation in the g.47918G>A SNP resulted in the change of codon from GAT to AAT, and the coding amino acid from Asp to Asn.The result of association of SNPs with slaughter traits showed various results were as follows:g.47915G>A and g.47918G>A had affected the eviscerated percentage.This result revealed that the polymorphism of MEF2A gene had basilic influence on slaughter traits.

To investigate the relationship between the expression of SYK and dairy cow mammary gland development and lactation, the expression of SYK in lactating dairy cow mammary gland with high or low quality milk and dry period Holstein dairy cow mammary gland was detected by Western blotting and laser confocal microscope.The results showed that SYK expression in dry period mammary gland was significant higher than that in lactating mammary gland (P<0.05).There was no SYK differential expression detected between lactating mammary gland with high quality milk and low quality milk (P>0.05).SYK was mainly located in the cytoplasm of ductal epithelial cells in dry period mammary gland.In lactating mammary gland, SYK was existed in acinar epithelial cells.All these results revealed that SYK was a regulator in mammary epithelial cell proliferation and differentiation.It participated in mammary gland reconstitution in dry period.

To study the correlation and regression analysis of body size, cashmere traits and economic traits in Inner Mongolia Erlangshan cashmere goat, we selected the 1 000 3-year-old ewe and carried out production performance measurement, and using SAS 9.0 and Excel 2003 softwares for statistical analysis.The results indicated that a significant positive correlation of body height(X₁), body length(X₂), heart girth(X₃) and weight(Y)in Erlangshan goat.Regression model was Y=-12.0856+0.0821X₁+0.4053X₂+0.1624X₃.The intercept and three partial regression coefficient had reached a very significant extent (P<0.01), and indicating the equation was the optimal regression equation.The heart girth was the biggest influence on body weight, and the body height, body length and cashmere fineness impacted on body weight through the heart girth.The body height was biggest influence on cashmere yield and cashmere thickness for the second.We did not find optimal regression model between body height, body length, heart girth, cashmere fineness, cashmere thickness, hair long and cashmere yield.These results indicated the presence of complex intrinsic link between body size, cashmere traits and economic traits in Erlangshan cashmere goat.

The persent study was aimed to investigate the optimization of mice cage time after superovulation, and efficient acquisition of mouse hatched blastocysts.The experiment selected 150 ICR female mice aged 8 weeks, were randomly divided into 5 groups, the same time superovulation treatment according to 1:1, after the male and female in 18:00, 19:00, 20:00, 21:00 alloy cage overnight, the next day as early as 08:00 check, found that vaginal suppository for the first day of the pregnancy (D1).Take the pregnant the fifth day (D5) mice were sacrificed, their bilateral uterine horns, rushed from the embryonic.Statistics each thrust ratio and the total number of embryos hatched blastocysts/take not hatched blastocysts, was used as the index to evaluate embryos to obtain efficiency;Statistical trophectoderm cell number/inner cell mass number, as a reference index to evaluate the quality of embryo.The results found that groups of Ⅰ, Ⅱ and Ⅲ were no significant difference in the blastocyst number (P>0.05), but there was increasing trend, group Ⅳ was significantly higher than the other three groups (P<0.05).Groups Ⅰ, Ⅱ, Ⅲ and Ⅳ within the cell mass cells number/trophectoderm cell number were 23.18%, 23.55%, 21.72% and 23.28%, there was no significant difference in each group (P>0.05).The results showed that the corresponding set of Ⅳ cage got the time of mouse hatched blastocysts to obtain the highest efficiency, there was no significant differences in embryonic blastocyst quality.

Reproductive development is related to the survival and thrives of one species.The female mammalian germ cell, that is oocyte, will obtain their reproductive capacity through two processes:Oogenesis and follicle development.These two interdependent processes will provide the foundations for the combination of male and female gametes, and for the starting of a new generation of ontogeny.With the development of molecular biology techniques, many regulatory mechanisms of gene expressions, which associated with follicle development, is revealed gradually.New ideas have emerged.This present review will summarize the major molecular pathways, which occurred at different mammalian follicle development stages.And these summarizations will provide theoretical basis for the studies of germ stem cells, development of therapeutic cloning, and the treatment and research of human reproductive diseases.In addition, the paper will also show the guiding significance to the acceleration of development of animal husbandry, utilization of early oocytes resources, shortening of the generation interval and improvement of reproductive rate.

There were various signal transduction pathways in cells.The interaction of different signaling pathways formed a complex signal transduction system, which involved in various physiological and biochemical functions of animals, and part of them played an important role in the animal reproductive physiology.Mitogen-activated protein kinases was one of the important signaling pathways in cells, which involved in the regulation of a variety of physiological processes.Especailly, they played an important role in animal reproduction.The article will summarize the roles of MAPK signaling pathways in animal reproduction.

Gram-positive cocci were isolated from half a month diseased lambs with a fatal infection in a sheep farm in Minhe county, Qinghai province.The clinical symptoms observed were drooping, reduced feed intake, emaciated and fever.The infected lambs died several hours after the disease been observed.Necropsy of two dying lambs revealed that for one lamb, the liver was enlarged and capsule of the liver dropped, and other organs had no obvious changes;For the other one, the organs had no obvious changes.After gram staining, gram-positive cocci were observed from all the samples.Partial Tuf gene sequences of these isolates were amplified and sequenced for species identification, BLASTn conparison result revealed that they had the highest sequence similarity with that of Enterococcus faecium (E.faecium) AUS0085 (99.8%), we thus primarily confirmed that these strains belonged to E.faecium.Antimicrobial susceptibility test indicated that these strains could be divided into two classes.Class 1 was resistant to β-lactams, aminoglycosides, macrolides, tetracyclines, quinolones, rifamycins, lincomycin and clindamycin;Susceptible to chloromycetin and vancomycin.Class 2 was resistant to β-lactams, aminoglycosides, tetracyclines, quinolones, rifamycins;Intermediate resistance to erythromycin and roxithromycin;But susceptible to clarithromycin, azithromycin, clindamycin, lincomycin, chloromycetin and vancomycin.Multilocus sequence typing (MLST) based on seven housekeeping genes for E.faecium revealed that E.faecium lby1 strain in class 1 and E.faecium lbg3 strain in class 2 were defined as a new sequence type ST989.Phylogenetic tree analysis revealed the representative strain of these isolates, E.faecium lbg3, was clustered together with E.faecium ST468.The pathogenicity test revealed that E.faecium lby1 and lbg3 strains both had pathogenicity.

The objective of the present investigation was to examine seroprevalence of Chlamydia in dairy cattle in Gansu and Ningxia areas, Northwest China, and to analyze the factors affecting Chlamydia infection in dairy cattle.Indirect hemagglutination assay (IHA) was used to detect antibodies against Chlamydia in 1 657 dairy cattle serum samples from Gansu and Ningxia areas.Epidemiological investigation and statistical methods were used to analyze the data.The results showed that the overall seroprevalence was 29.33%, logistic regression analysis was applied to evaluate the risk factors of Chlamydia infection in dairy cattle, and the results indicated that age and numbers of pregnancy of dairy cattle were not the significant risk factors (P>0.05), and were left out of the final model, however, region was considered as the main risk factor associated with Chlamydia infection (P<0.05).The highest titer was 1:1 024.In conclusion, the results of the present survey indicated the widespread of Chlamydia infection in Gansu and Ningxia areas.In order to ensure the economic benefit of dairy farming, we should pay more attention to Chlamydia infection in dairy cattle, and integrated control strategies and efficient management measures should be implemented to prevent and control Chlamydia infection in dairy cattle.

To explore etiology characteristics of Staphylococcus sciuri, strain YDE0916 was isolated and identified from Corydoras by cultivation, biochemical identification, 16S rDNA gene PCR amplification and sequence analysis, and resistance analysis.The result showed this strain was gram-positive, serial, spore, without capsule.The physicochemical properties showed immobility, β-galactosidase-negative, VP reaction-negative, sugar and maltose-positive.The nucleotide sequence of 16S rDNA was 1 476 bp and its GenBank accession number was KP031644.This 16S rDNA showed 99% homology with Staphylococcus sciuri.Phylogenetic tree showed that strain YDE0916 and several Staphylococcus sciuri strains gathered into one cluster.So the strain was identified as Staphylococcus sciuri.The antibiotic sensitive test showed that strain YDE0916 was resistant to norfloxacin and bacitracin, and was highly sensitive to 22 kinds of drugs, such as minocycline, gentamicin and ampicillin.This study could provide a reference for identification and prevention of Staphylococcus sciuri.

In order to study the effect of different test materials on the detected results of avian leukosis virus (ALV)-p27 antigen by ELISA, the cloacal swabs ALV-p27 antigen were detected by ELISA in 180-day local breeds hen, and the parts of the positive and negative chicken was selected to group test, the serum, egg whites and cell supernatant ALV-p27 antigen were detected by ELISA, the specific genes of ALV were detected by RT-PCR in blood.The results showed that serum, egg white and cell supernatant as ELISA test materials, the positive rate lower of than cloacal swabs, and serum ALV-p27 positive samples include all egg whites and cell supernatant positive samples in positive group.It was a significant correlation between ELISAs with serum and cell supernatant (linear equation:Y=1.8439X-0.1469, the correl was 0.937).In positive group, ALV-p27 gene positive rate lower than cloacal swabs ELISA, but higher than the serum, egg and cell culture medium, and ALV-p27 gene positive samples include serum positive samples by ELISA.ALV-J gp85 gene positive rate of 29.17%, and all positive samples were included in the serum ALV-p27 positive samples.The results suggested that the cloacal swabs as test material may occur false positive results, and egg whites and cell supernatant may occur undetected by ELISA ALV-p27 antigen assay in adult chicken, serum as test material in ELISA could more accurately reflect the state of adult chickens infected with ALV.

This study was aimed to provide the basis for prevention and control of development of a new product for bacterial respiratory disease of chicken.Diseased samples such as throat, trachea and lungs were inoculated on various selected media for isolating bacteria.17 isolated strains were identified by biochemistry assay and PCR, and then animal experiment were carried out.Black and glassy surface with metallic shine colony from EMB medium Gram-negative bacilli were observed under optical microscope.1 500 bp products were specifically amplified by PCR.DNA sequencing results were analyzed by BLAST and DNAStar with 16S rDNA sequence in GenBank.The results showed that the strains were identified as Escherichia coli.It could cause damage infection with the isolated bacteria through noses.Significant blood spots were observed on throat.Trachea was congestion, swelling and studded with blood points.Heart and liver were wrapped with yellow cheese substances.Air sac was yellow and opaque.The isolated bacteria could provide basis for developing a new product.

In order to provide a scientific prevention and treatment of Haemophilus parasuis (Hps) disease in pig farms of the Tianjin area, pigs with suspected Haemophilus parasuis disease were anatomized and observed pathological changes.The most common results were cellulose pleurisy and peritonitis, and there were more yellow viscous liquid in joint cavity section.The bacteria were isolated from 36 diseased pig swine organs which were suspected of Hps, and 9 strains of Hps were obtained through staining, satellite test, catalase reaction test, biochemical reaction and PCR.The 9 strains of Hps showed multi morphology and gram negative through staining; Satellite phenomenon was significant; All of the catalase reaction test were positive; D-ribose, sorbitol, fructose, arabinose and hydrogen sulfide of micro biochemical reaction were negative, but maltose, glucose, sucrose, mannitol and urease were positive, only 1 of 9 strains was positive sorbitol and xylose of micro biochemical reaction, 5 strains were positive for beta galactose;PCR result showed an objective fragments of 822 bp.The separation rate was 25%.The drug screening were done with 16 kinds of antimicrobial agents against 9 strains of Hps, drug sensitivity results showed that ceftriaxone sodium and levofloxacin had good antibacterial effects.The results of this study could provide the basis for prevention and control of Hps disease in Tianjin.

Productive disorder induced by porcine reproductive and respiratory syndrome virus (PRRSV) is characterized by low conception rate, irregular return to estrus and abortion during early gestation sow; Later-term abortion, early farrowing, stillborn, mummies and weak-born piglets are generally happened at second half of gestation.Piglets with congenital infection become more sensitive to secondary infection, and there exists excreted virus which infects the negative animals on every side.PRRSV passes through the placenta mediated by macrophages/monocytes, which cause pathological damage in the maternal-fetal interface, and virus replication in the fetal implantation sites plays a role in the fetal death and sow abortion.PRRSV infection prevention and viremia shortening for sow are conductive to reduce PRRSV vertical transmission.In addition, biosafety measures, reasonable pig flow and strict disinfection are carried out to control horizontal transmission, and vaccine immunization and medicine administration are done to protect susceptible animals.The main purpose of this review is to summarize PRRSV vertical transmission during each stage of gestation and subsequent reproductive failure.

In recent years, canine distemper is a serious damage to health development of dog and fur animal breeding industry.Fast and accurate diagnostic technique is particularly important for canine distemper prevention.At the present stage, canine distemper diagnostic techniques mainly include virus isolation, culture and microscopy observation, enzyme-linked immunosorbent assay(ELISA), immunohistochemistry, immune colloidal gold technique, nucleic acid hybridization technique, nested RT-PCR, Real-time fluorescence quantitative RT-PCR, double PCR, gene chip technology, liquid phase chip technology, loop-mediated isothermal amplification technology, and so on.The key of isolation, culture and microscopy observation of canine distemper virus is to obtain a susceptible cell lines that can produce typical CPE.We review research progress on diagnostic techniques of canine distemper, so as to provide simple and highly effective detecting technique for preventing and controlling epidemic and outbreak of canine distemper.