The present study was aimed to explore the potential role of Toll-like receptors (TLRs) on the mink immunity, and accumulate alternative genetic material for the mink breeding for disease-resistant.Four pairs of primers were designed according to ferret TLR sequences in GenBank to obtain the sequences of TLR genes (TLR4, TLR6, TLR7 and TLR8) of mink and then performed bioinformatics analysis of these sequences.In addition, the expression of TLR4 and TLR7 genes in various tissues in young and adult minks were analyzed by RT-PCR. Sequence homology analysis showed that mink had higher homology with carnivora (ferrets, polar bear, panda, walrus, seals, dog, tiger and cat) which had the nearest perimeter in the phylogenetic tree.Tissue expression analysis showed that TLR4 and TLR7 were widely and differently expressed in variety tissues of mink.Interestingly, the expression of TLR4 and TLR7 in young mink were much higher than that in adult mink.

Interleukin-6 (IL-6) is an important immune regulatory factor, and has great development prospects in the application of adjuvants.Using the Banna minipig inbred line (BMI) as the experimental animal, we successfully cloned the coding region of pig IL-6 gene through polymerase chain reaction (PCR) method, and had deposited the sequence into NCBI database and assigned to the accession No.JQ839263.Then we further studied this gene by bioinformatics analysis method.The results showed that BMI IL-6 encoded 212 amino acids with a predicted molecular weight of 23.88 ku, the isoelectric point (pI) was 6.66 and a signal peptide in N-terminal.It was a nuclear protein with 89% probability.In addition, BMI IL-6 contained one conserved structure domain and one transmembrance structure, and its N-terminal and C-terminal were both hydrophobic.Comparing the sequence of the coding region of BMI IL-6 gene with the download 4 IL-6 gene sequences of pig, the result showed BMI was identical with GenBank accession No.NM_214399 and DQ832259, only had one base pair difference with GenBank accession No.AF309651 and AF518322, respectively.Multi-species amino acid sequence alignment and phylogenetic analysis demonstrated that BMI IL-6 showed the closest relationship with camel.Finally, we analyzed IL-6 gene mRNA expression in 12 important tissues of BMI through PCR method, the results showed that BMI IL-6 gene was highly expressed in lung, skin and muscle of BMI.These data laid a foundation for further insight into the expression and regulation of this gene of pig.

In order to establish a duplex loop-mediated isothermal amplification (LAMP) method for the detection of shellfish parasitic infection with Perkinsus and Bonamia, primers were designed according to the conserved region of ITS and 18S rRNA gene sequences of Perkinsus and Bonamia deposited in GenBank using the online Primer Explorer 4.0.Firstly, the respective single LAMP detection assay of two parasites were developed, and then the reaction conditions were optimized by combining the two reaction systems into duplex LAMP method.The specificity and sensitivity of duplex LAMP method were examined and the confidence was evaluated by digesting the amplicons with respective endonuclease.The results showed that the established duplex LAMP method was specific enough to distinguish the two parasites from other parasites, and the detection limit of Perkinsus was 10 copies/μL plasmid DNA, the detection limit of Bonamia was 100 copies/μL plasmid DNA.The endonuclease digestion of duplex LAMP products certified the confidence of the established duplex LAMP method.The detection result of 20 Ruditapes philippinarum samples collected randomly from the East China sea and Yellow sea and 10 Ostrea edulis samples imported from USA showed that the duplex LAMP method could be used as a rapid detection method for infection with Perkinsus and Bonamia.

The morphological features of Haplorchis pumilio (H.pumilio) and H.taichui were observed with hydrochloric acid carmine staining; The ITS2 sequences were amplified by PCR, analyzed by sequencing, and compared with sequences of H.yokogawai and Metagonimus yokogawai of Heterophyidae trematodes downloaded from GenBank; And the phylogenetic tree was constructed by Mega 6.05.The morphological characteristics of H.pumilio and H.taichui from dog and cat were conformed to the classical taxonomic description.The ITS2 sequences of H.pumilio and H.taichui were 295 and 297 bp, and the percentages of G+C content of ITS2 were 49.5% (146/295) and 54.2% (161/297), respectively.The sequences were submitted to GenBank and the accession numbers of H.pumilio were KJ137221-KJ137223 and KP165437-KP165439, the accession number of H.pumilio was KP165440.The sequence alignment results showed that H.pumilio and H.taichui intraspecific difference was 0, interspecific differernce with four heterophyes was 15.2% to 28.9%.The phylogenetic tree based on ITS2 showed that H.pumilio and H.taichui were at the same clade with the bootstrap value of 99%.

One pair of primers had been designed and synthesized based on the α-toxin gene of Clostridium perfringens.The complete α-toxin gene fragment was amplified by polymerase chain reaction (PCR), and then was cloned into pGEM-T Easy vector to construct pGEM-T-α.Digested with EcoRⅠ and Hind Ⅲ, a fragment of 1125 bp was cloned into the expression plasmid vector pET-28a(+).The recombinant plasmid was transformed into the BL21(DE3)plys and induced by 1.0 mmol/L IPTG at 37 ℃.The expression product was found to be 46.1 ku as expected one identified by SDS-PAGE, and confirmed by Western blotting with Clostridium perfringens type A antisera, indicating similar reactivity with native α-toxin.Recombinant α-toxin protein was simultaneously found in culture supernatant, postsonic supertanant and inclusion bodies, most protein was expressed in inclusion bodies, which indicated recombinant α-toxin protein was expressed in the extracellular, periplasm and cytoplasm.Recombinant α-toxin protein in postsonic supertanant could not make mice die, indicating its non-toxicity.Toxin-antitoxin neutralization test showed that antisera of recombinant α-toxin protein were specific to α-toxin.Upon immunization of rabbit with the recombinant α-toxin protein, antisera with high antibody titer neutralizing 100 MLD toxin per 1 mL were prepared.

In order to research the biofunction of porcine cyclin dependent kinase 2 (pCDK2), this study prepared polyclonal antibodies against pCDK2.The recombinant prokaryotic expression plasmid pET28a-pCdk2 was constructed and transformed into BL21 (DE3), IPTG was used to induce the expression of recombinant pCDK2 (rpCDK2) which was then analyzed by SDS-PAGE and Western blotting.The expressed proteins were purified and emulsified with freund's adjuvant, the mixture was used to immunize the rabbits to prepare polyclonal antibodies against pCDK2.The immunocompetence of the antibody was identified by immunoperoxidase monolayer assay (IPMA), and the specificity was confirmed by Western blotting.The results showed that rpCDK2 was successfully expressed by prokaryotic expression system, and the expression products showed two forms:Solubility and inclusion body(IB), the molecular weight of the solute rpCDK2 was about 38 ku, however three different molecular weight forms (from 38 to 43 ku) of IB were observed.IPMA results showed that polyclonal antibodies against pCDK2 protein showed perfect activity which could recognize pCDK2 expressed in PK-15 cells and ST cells specifically.Western blotting analysis showed that four bands with different molecular weight forms (from 34 to 50 ku) were observed when the polyclonal antibodies against pCDK2 protein reacted with the total protein extracted from PK-15 and ST cells.In conclusion, the rpCDK2 protein was successfully expressed by prokaryotic expression system, the prepared rabbit polyclonal antibodies against pCDK2 protein showed wonderful activity and specificity, and the study provided basic material for the research of protein biofunction and related diseases.

In order to investigate the epidemiology and genetic variation of avian infectious bronchitis virus (IBV) in Guangdong province, two strains of IBV were isolated by inoculation of embryo and RT-PCR detection from diseased chickens at different farms in Zhanjiang, Guangdong province in 2013.We denoted these two strains of IBV as CK/CH/GD/ZJ10/2013 and CK/CH/GD/ZJ11/2013, respectively.Analysis of the S1 gene sequences from these two isolated strains showed that the S1 gene of CK/CH/GD/ZJ10/2013 was 1 626 bp, which encoded 542 amino acids with a cleavage site sequence of NRFRR, while the S1 gene of CK/CH/GD/ZJ11/2013 was 1620 bp, encoded 540 amino acids with a cleavage site sequence of HRRRR.Phylogenetic analysis revealed that these two isolated strains were clustered into genetic groups Ⅲ and Ⅰ(QX-like type), respectively.Homology analysis demonstrated that nucleotide and deduced amino acid sequence homologies between these two isolated strains were lower, while the homologies were higher among the same genotypes, however, the homologies were lower comparing with current vaccine strains such as H120 and H52, with which the nucleotide homologies ranged from 75.7% to 76.3% and the amino acid homologies ranged from 77.1% to 77.9%.Further analysis showed that the CK/CH/GD/ZJ10/2013 strain was formed from a recombination event between CK/CH/GX/NN11-3 and GX-NN-6.Taken together, this study provided valuable insight into prevention and control of IBV infection in Guangdong province.

To explore the molecular characteristics of protein A6 from goatpox virus (GTPV), genomic DNA was extracted from goatpox virus Gulang isolate.The specific primers were designed and used to amplify the A6L gene from the genomic DNA by PCR.A PCR product was ligated into pGEM-T Easy vector.After transformation into E.coli DH5α, the positive clones were sequenced and the sequences were analyzed with the bioinformatics softwares.The result showed that A6L gene sequence contained an open reading frame (ORF) of 1128 nucleotides and deduced protein consisted of 375 amino acids with the theoretical molecular weight of 43.68 ku and isoelectric point of 7.50.Analysis of secondary structure of protein A6 revealed that α-helix, β-strand and random coil were 53.30%, 10.24% and 39.46%, respectively.Analysis of multiple sequence alignment indicated A6L gene from different capripoxvirus isolates were highly conserved with identity of more than 97%.The present results laid the foundation for further studies of biological functions of protein A6 and interaction among the early proteins of capripoxvirus.

To develop a detection method for rapid diagnosis of dairy cow tuberculosis, evaluation of the raw milk contamination status and tracing the route of transmission, specific primers of Mycobacterium bovis were designed to develop the FQ-PCR assay and the reaction conditions were optimized.Standard curve of the FQ-PCR test was developed and its properties were evaluated.The results showed that the developed FQ-PCR test could be used to detect the Mycobacterium bovis.The best primer concentration and annealing temperature were 400 nmol/L and 52 ℃, respectively.Properties evaluation showed that the method had good specificity, sensitivity, repeatability and clinical application.The test results indicated this method could be used to qualitatively and quantitatively detect Mycobacterium bovis.It would be an important technology for diagnosis and decontamination of dairy cow tuberculosis and safety assessment of raw milk.

The research was designed to analyze the rule of mRNA expression of IgG Fc receptor(FcR) in intestine from Yili horse aged at two-years-old by RT-PCR method, which aimed at providing data for researches on immunology mechanism and digestive physiology of equine, improving immunology function of horses, enhancing of nutrients absorbing, decreasing mortality.The results showed that the expression of IgG FcR gene in small gut was significantly higher than that of cecum (P< 0.05);it was also higher than that of large gut, its expression in large gut was higher than that of cecum(P> 0.05).The expression of IgG FcR had been identified in intestine of horses at two-year-old, it was different of immunology mechanism among horses, human and rat;Moreover, the expresson of IgG FcR in small gut was the highest among intestine, its expression rule was similar as other mammal animals (piggy and sheep).Above all, not only could be small gut thought as a digestive and absorption organ in nutrients, but also a important immunology organ in terms of its function, each plays special role in physiology of animal body.

This study aimed to construct Oct4-EGFP pluripotent reporter vector in pig without destroying cells researching the expression pattern of Oct4, thus contributing to early embryonic development and stem cell research.Construct Oct4-EGFP pluripotency reporter vector with the In-Fusion PCR cloning technology, the Oct4 promoter sequence to direct the recombinant vector pEGFP-N1 instead of EGFP original CMV promoter plasmid pEGFP-N1 by Oct4 and liposomal transfection technology, and transfected into livability of porcine fetal fibroblasts cells.The results showed that it had successfully constructed Oct4-EGFP pluripotency reporter vector by PCR and sequencing verified, and initially verified the validity of the carrier in the parthenogenetic blastocysts.After liposomal transfection, 8 genetically modified cells of incorporates pluripotent report carrier with the Oct4-EGFP had been gained by screening and PCR.Thus, the In-Fusion PCR cloning technology could efficiently construct Oct4-EGFP pluripotency reporter vector and obtained transgenic positive cells could lay the foundation for the early development and embryonic stem cell research of pig embryos.

To obtain VP7 protein of bluetongue virus (25 type), VP7 gene was amplified and cloned in pET-24b(+) expression vector.The pET-24b-BTV-VP7 recombinant plasmid was transformed into BL21 (DE3), then the VP7 protein of bluetongue virus was expressed using IPTG and purified by nickel affinity chromatography in vitro.Immunogenicity of VP7 protein was determined by Western blotting and ELISA.The results showed that the molecular weight of VP7 protein was about 40 ku and it could react with goat positive serum specifically.This study laid the foundation for establishing protein chip detection methods in the future.

The purpose of this study was to analyze the conservative of heat shock protein 90(HSP90) gene, and provide theoretical data for the new HSP gene by exploring the application of HSP90.Using CODEHOP to design the degenerate primers, cloning Carassius auratus HSP90 sequence by PCR, and then the sequence was cloned into pMD18-T vector to construct clone vector after sequencing.Then sequencing homology were compared by BLAST and phylogenetic tree was constructed.The results showed that the homology was 99% between Carassius auratus and Cyprinus carpio; And the homology was 92% between Carassius auratus and Danio rerio.The degeneracy of primers designed in this study was low, the modification of Tm value was more flexible, and the product had high specificity.The study provided basis guarantee for research of Carassius auratus function genes, and laid the theoretical foundation for the study of heat shock proteins conservative in evolution.

This experiment was conducted to study the effects of pasting anionic salt and RPC on production performance and blood biochemical indexes of perinatal dairy cows.Using randomized trial design, 20 healthy Holstein cows which according to the tire time (2 to 4), age, last time lactation quantity and parturition date were chosen.They were divided into four groups, 5 cows per group.The control group was fed TMR diet, Ⅰ, Ⅱ, Ⅲ groups were fed the basal diet +200 g pasting anion salt and supplemented with 10, 20, 30 g/(head·d) RPC, respectively.There were 7 days to pilot test, formal test was from antenatal 21 d to parturition.The results showed that:①DMI of all experimental groups were significantly higher than that of control group during most of antenatal 21 days (P< 0.05);The nutrients apparent digestibility of all experimental groups were generally higher than that of control group;Add 20, 30 g/(head·d) RPC was significantly increased the apparent digestibility of CP, NDF, ADF (P< 0.05);The apparent digestibility of Ca of all experimental groups were significantly higher than that of control group (P< 0.05);Add 30 g/(head·d) RPC was significantly increased the apparent digestibility of EE (P< 0.05).②Among postpartal 90 days, the total milk yield of all experimental groups were significantly or extremely significantly higher than that of control group (P< 0.05;P< 0.01);Add 20, 30 g/(head·d) RPC was significantly enhanced the feed conversion rate (P< 0.05);Add 30 g/(head·d) RPC was improved the percentage of milk dry matter, milk protein and milk fat of colostrum (P< 0.05).③Serum Ca²⁺ concentration of all experimental groups were significantly higher than that of control group (P< 0.05);Add 20, 30 g/(head·d) RPC was significantly increased the part time plasma TG levels (P< 0.05), significantly reduced the part time plasma contents of TCHO, BHBA and NEFA (P< 0.05).In conclusion, among the perinatal period added pasting anionic salt and RPC could improve early prenatal DMI and part nutrients apparent digestibility, improve perinatal blood calcium and blood lipid metabolism, and enhance the production performance and feed conversion rate, based on adding 200 g pasting anion salt among early perinatal, the best add amount of RPC was 30 g/(head·d).

The objective of this experiment was to study the effects of production of white veal with Holstein bull calves fed on the liquid milk replacer (containing mainly dairy products) feeding and the dry milk replacer (containing mainly plant diet, etc)feeding.20 healthy male calves with similar birth date and body weight were selected, and randomly divided into two groups (10 calves per group), for a period of 120 d white beef production fattening test.GroupⅠfed with milk replacer (commercial, 22% CP, 16% EE), groupⅡ fed with milk replacer (formulated, 22% CP, 16 MJ/kg DE).The results showed as follows:① 0 to 90 d ADG of group Ⅱ were higher than groupⅠ, 91 to 120 d ADG of group Ⅱ were significantly lower than groupⅠ (P< 0.05), but there were no significant differences in 0 to 120 d ADG and 120 d weight between two groups (P> 0.05).② There were no significant differences in the nutrient apparent digestibility of Holstein bull calves with different milk replacer and feeding methods (P> 0.05).③ Carcass weight, dressing percentage and eye muscle area of groupⅠcalves were significantly higher than those of group Ⅱ (P< 0.05), increased by 11.88%, 11.04% and 8.12%, respectively, but there was no significant difference in carcass meat production rate between two groups (P> 0.05).There was no significant differences in pH, shear force, cooking lose and meat color of longissimus between two groups (P> 0.05), but compared with group Ⅱ, the water lose rate of groupⅠincreased by 20.83% (P< 0.01).The content of EE of groupⅠwas significantly higher than group Ⅱ (P< 0.01), but there were no significant differences in the contents of moisture, CP, Ash, Ca, P and Fe of longissimus between two groups (P> 0.05).④Compared with groupⅠ, the feed cost of group Ⅱ was reduced by 45.38%.In summary, the growth performance and meat quality of two groups calves were similar, feeding cost was reduced significantly.

The experiment was designed to study the effect of different strain combination on the nutricines content and in vitro digestibility of solid state fermentation material.In this experiment, the solid state fermentation media included wheat bran, corn bran, corn meal, rice bran, etc.The Saccharomyces cerevisiae (BC, XR4) and Bacillus subtilis (A15) were inoculated on the solid state fermentation media in different proportion combination.Group 1 was inoculated with BC and XR4, with a ratio of 1:1 and group 2 was inoculated with BC, XR4 and A15, with a ratio of 1:1:0.5.While the control group was without strains.By measuring the content of 5 nutricines (β-glucan, mannan, peptides, amino acid and organic acid) and in vitro digestibility of DM, CP, NDF and ADF in fermentation substrate to evaluate the effect of different strain combinations on solid state fermentation quality.The results showed as follows:① The nutricines content of 3 groups increased with the extension of fermentation time.Among them, the group 2 had an optimal fermentation effect, compared with the pre-fermentation, the contents of β-glucan, mannan, peptides, total amino acid, total organic acid were increased by 27.69%, 44.72%, 27.62%, 9.01% and 474.99%, respectively.② In vitro digestibility of CP, NDF and ADF in groups 1 and 2 were significantly higher than that in control group (P< 0.05), compared with the control group, the in vitro digestibility of DM, CP, NDF and ADF in groups 1 and 2 were increased by 0.15% and 4.44%, 3.29% and 4.43%, 10.37% and 23.00%, 33.04% and 131.59%, respectively.Group 2 was better than group 1.Thus, on the basis of Saccharomyces cerevisiae (BC and XR4), adding Bacillus subtilis (A15) would bring best fermentation and it not only could significantly improve the nutricines content, but also could significantly increase its in vitro digestibility.

The aims of this study were to understand the relationship between the indexes of body size and body weight in Holstein cows, and the variation pattern of body size and body weight of different age stages of Holstein cows in Changji area of Xinjiang were analyzed.This study based on the measurement record of the body size and body weight of 2 972 Holstein cows of different age stages from 2012 to 2013 in Changji area of Xinjiang, obtained the body size index of different age stages, and mapped out the cumulative growth curve, absolute growth curve and relative growth curve;At the same time, carried on a correlation analyses between each body size index and body weight of Holstein cows.The results showed that along with the increase of age, body size index and body weight of Holstein cows were increasing;At the age of 1 to 3, the growth and development were rapid, it was the most important period for the growth and development of cattle.The correlation coefficients among body height, body length, chest circumference, cannon circumference and body weight were reached significant level (P< 0.01).It was important to pay special attention to breeding management of Holstein cow before three years old, and ensured its nutritional requirements in order to achieve the best physical condition in the process of breeding.

This study was aimed to compare the meat quality of Gayal×Brahman (GBF₁) cattle and Yunling cattle, evaluate the potential of crossbreeding utilization of Gayal.GBF₁ cattle and Yunling cattle which had same ages and same gender were selected to be fattened and slaughtered.100 meat samples of GBF₁ feedlot steers and Yunling feedlot steers were collected and sealed in clean fresh frozen bag.With the meat samples from Yunling feedlot steers as the control, the shear force, lose water ratio, water holding ratio and cooking loss of meat samples from GBF₁ were determined.The results showed that the shear force of shoulder crest from GBF₁ cattle were significantly higher than that from Yunling cattle (P< 0.05), and there were no significant differences in the others meat samples (P> 0.05).The average shear force, average lose water ratio, average cooking loss of 10 different cuts from GBF₁ cattle were 3.50 kg, 24.15%, 27.56%, respectively and lower than that of Yunling cattle (P> 0.05).The average water holding ratio was 64.60% and higher than that of Yunling cattle (P> 0.05).In conclusion, the meat quality of GBF₁ cattle was good, similar to that of Yunling cattle.

This experiment objective was to study the Fe, Cu, Mn, Zn, Se five kinds of trace element contents, distribution characteristics and mutual relations in the soil-grass-water (stream) in Yili pastoral area of Xinjiang, to evaluate the trace elements nutritional-ecologic environment of Yili summer pasture in Xinjiang.At the altitudinal belt of 1400 to 2999 m of the summer pasture of the Karajon grassland (South slope) and Tangbula grassland (North slope) of Yili, Xinjiang, the samples of soil (n=75), herbage (n=75), stream water (n=75) were collected according to the altitude, to determine their contents of Fe, Cu, Mn, Zn and Se for evaluating the nutritional-ecologic environment of trace elements of Yili summer pasture.The results showed that the contents of Fe, Cu, Mn, Zn in the soil of summer pasture on average were 15 418.4, 32.6, 474.5 and 115.8 mg/kg, respectively;And 446.0, 7.8, 85.7 and 41.9 mg/kg, respectively in forage;The correlation (R²) of the contents of Fe, Cu, Mn and Zn between soil and forage were 0.43, 0.82, 0.47 and 0.72, respectively;The changes of Fe, Cu, Mn and Zn contents in the soil and forage were not significantly related with the altitude.There were no the 4 elements mentioned above in river streams, there was no Se in the soil and forage, and there was infinitesimal of Se in the river streams.It was concluded that in Yili summer pasture there were excessive Fe, suitable Zn and Mn, insufficient Cu, and more insufficient Se.

The recently discovered nesfatin-1 is a new anorexigenic molecule.The precursor protein nucleobindin-2 (NUCB2) is proteolytically cleaved into three fragments, nesfatin-1, nesfatin-2 and nesfatin-3.The middle segment of nesfatin-1 (M30) is responsible for inhibiting food intake, while the physiological role of nesfatin-2 and nesfatin-3 are not currently known yet.nesfatin-1 plays roles in behaviour, mood, reproduction, sleeping, epilepsy and diabetic hyperphagia.We reviewed the discovery, designation, biochemical structure, distribution, its main biological function and the possible mechanism of inhibiting food intake effect of nesfatin-1.

To study the optimal condition of fermented soybean meal production, several factors were selected to study the effects of different strains inoculation quantity, fermentation temperature, moisture content, fermentation time and accessories on soybean meal fermentation.The results showed that when Bacillus subtilis, Saccharomyces cerevisiae and Lactobacillus addition proportions were 3‰, 2‰, 1‰, respectively, proteinase supplementation was 2‰, strains activation time was 0.5 h, initial moisture content was 38%, fermentation temperature kept at 30 to 42 ℃, physical and chemical indexes of fermented soybean meal were optimal, the contents of small peptide and lactic acid could reach more than 20% and 3.5%, respectively, and at the same time, quality of fermented soybean meal was very stable.

This study was attempted to generate one Marc-145 cell line stably and highly expressing porcine CD163 (pCD163) and set the foundation for PRRSV isolation and vaccine production.CD163 was shown to be a cellular receptor capable of mediating infection of PRRSV non-permissive cell lines.The pCD163 gene was amplified by RT-PCR from porcine alveolar macrophages and cloned into the eukaryotic expression vector pCI-neo, then the positive plasmid pCI-pCD163 was transfected into Marc-145 cells.After selecting with G418 and subcloning for 3 times, Marc-145 cell line expressing pCD163 was established.IFA results indicated that the fluorescence of pCD163-Marc cells was significantly brighter than Marc-145 cells;Western blotting results indicated that the pCD163-Marc cells could express higher levels of CD163 and the expression level was 8.7 times higher than Marc-145 cells.The pCD163-Marc cell line could be stably passaged for 20 passages and the expression level of CD163 was similar with different passages, which would be a valuable tool for facilitating virus propagation and vaccine production.

The enzyme was efficient active gene product of catalyzing metabolic pathway.Stress environment and genetic basis led the differences of isozyme structure or activity, which specific changes of the kinds and activities of enzyme were showed in different tissues and developmental stages.Here, lactate dehydrogenase (LDH), alcohol dehydrogenase (ADH), peroxydase (POD) and catalase (CAT) were respectively extracted from brain, heart, liver, lung and spleen tissues and organs of goslings infected by goose parvovirus (GPV) and control group.The results of PAGE and biochemical analysis showed that there respectively were 3 and 1 slow-zone CAT isozyme bands deletion in liver and spleen of goslings infected by GPV.There respectively were 1 new enzyme band of POD isozyme in the brain, heart and spleen of goslings infected by GPV, but 1 enzyme band of POD isozyme deletion in the lung of goslings infected by GPV.It only showed 1 to 2 slow-zone LDH and ADH isozyme bands in organ and tissue of goslings infected by GPV, and deleted 1 slow-zone ADH isozyme band in the lung of goslings infected by GPV.The activity of ADH from goslings infected by GPV were reduced 13.61% to 61.08%, but increased 1.2 times in the heart of goslings infected by GPV.The activity of POD, CAT and LDH from goslings infected by GPV were higher 1.2 to 6, 1.5 to 3.5, 2 to 2.5 times than that of control group, respectively.The activity of CAT reduced 40.29% and 94.68% in the brain and lung of goslings infected by GPV.The activity of LDH reduced 75.93% and 91.81% in liver and lung of goslings infected by GPV.Those results indicated that the interaction of GPV stress and host oxidase/dehydrogenase enzyme gene expression resulted in the biochemical characteristics variation of activity and isozyme patterns of oxidase/dehydrogenase enzyme, and directly or indirectly affected metabolic approach and physiological pathological function on goslings infected by GPV.Therefore, sensitive oxidase/dehydrogenase was the effective marker of pathological mechanism on investigation of virus genotype interaction with genetic susceptibility of the host.

To understand the effect of intestinal digestion and absorption on the growth and development of lambs.60 5-month and healthy lambs were selected as the study subjects which were F1 generation hybridized by Altay sheep × Teksaier sheep and were fed in the same rearing conditions and had the same weaning date.And then, 8 lambs with significant weight differences were equally divided into HBW group and LBW group, the former with average weight (37.20±2.80)kg and the latter with average weight (28.24±3.75)kg.Measurements were to be performed for the differences in the morphological indicators of villus in duodenum, digestive enzyme activity in duodenum and sodium glucose cotransporter 1 (SGLT1) and glucose transporter 2 (GLUT2) in the lambs'mucosa.The results showed that the villus height and crypt depth of duodenum in HBW group were significantly lower than that in LBW group (P< 0.05), there were no significant differences in the mucosal thickness and V/C between two groups (P> 0.05);The amylase activity in HBW group was significantly higher than that in LBW group (P< 0.05), the activities of maltase, lactase and sucrase in HBW group were extremely significantly higher than that in LBW group (P< 0.01), there were no significant differences in the activities of lipase and protease between two groups (P> 0.05);The expression amount of SGLT1 in HBW group was significantly higher than that in LBW group (P< 0.05), the expression amount of GLUT2 in HBW group was extremely significantly higher than that in LBW group (P< 0.01).In conclusion, there was close correlation between the weight increment of the lambs and digestive and absorptive functions of duodenum, and that further research and study need to be conducted to have a better understanding of the regulating mechanism of molecular biology for digestive enzyme and glucose transporter.

Interleukin-2(IL-2) gene was regarded as a candidate gene for immune function of Jingxinghuang chicken in this study.The genotype frequencies, allele frequencies, haplotypes and association with H/L of five SNPs were studied by direct sequencing.The results indicated that all the five loci fitted Hardy-Weinberg equilibrium and showed low degrees of heterozygosity with two genotypes detected.Meanwhile, the homozygosis genotype was predominant in all the five loci.Analysis of linkage disequilibrium (LD) suggested that SNP1 and SNP2 showed perfect LD, whereas SNP3, SNP4 and SNP5 showed perfect LD.Analysis of haplotypes revealed four haplotypes, among which Hap1 (TGACT) was predominant.The least square method (LSM) analysis showed that there were no significant difference between different genotypes and haplotype combinations(P> 0.05).The results suggested that more SNPs of IL-2 gene and association with additional immune traits would be investigated in future studies for better understand the roles in immune systems that chicken IL-2 playing.

The aim of the present study was to explore the influence of both fasting and water deprivation stress on horses.Eight adult Yili gelding were used with a incomplete Latin square design.The animals were divided into fasting and water deprived for 49 h group (feed and water deprivation group, FW group) and normal feeding group (control group).The changes of basic vital signs, blood biochemistry indexes and stress-related hormones indicators were monitored during experiment from horses.The results showed that heart rate of FW group at 12 to 48 h were significantly lower than the control group (P< 0.05), breathing rate at 48 and 49 h were significantly lower than the control group (P< 0.05), blood glucose concentration at 18, 24 and 36 h were significantly lower than the control group (P< 0.05), plasma cortisol level at 18, 30 and 36 h were significantly higher than the control group (P< 0.05), plasma antidiuretic hormone level at 48 and 49 h were significantly higher than the control group (P< 0.05).There were no significant differences in other parameters between two groups (P> 0.05).The results suggested that horses could maintain basic life activities by reducing cardiovascular activity and body energy consumption, and improving the stress ability in the condition of fasting and water deprivation.

The bovine mammary epithelial cells not only have the function of synthesis and secretion of milk, but also play an important role in innate immunity system of the mammary gland, and there is great significance of them on studying the mechanism of lactation, mastitis pathogenesis and drug screening.Primary cultured bovine mammary gland epithelial cells are suitable for setting up cell model which can be used as the dielectric for physiological, pathological and pharmacological researching, avoiding the difficulties of in vivo test, such as the long cycle, the high cost, the individual difference, etc.The author summarized the latest researches of cell primary culture in vitro, cultivation technology, purification and identification method in order to provide reference for the studies of bovine mammary epithelial cells culture.

The experiment was aimed to study the effects of dietary energy level on body weights, serum testosterone concentrations and semen quality of Shaanbei White cashmere goat bucks.A total of 12 healthy bucks were selected and divided into 4 groups according to randomized block design with 3 replicates per group, each replicate included 1 buck.Four diets were formulated by the recommendation of NRC (1981, 2007) at 85%, 100%, 115% and 130%.The results showed as that the dietary energy level had extremely remarkable effect on bucks' ADG in each stage (P< 0.01).The weight of bucks in groups Ⅰ and Ⅱ decreased firstly then increased while there were a trend of increasing firstly then decreasing then increasing again in groups Ⅲ and Ⅳ.Serum testosterone levels and ejaculations under the frequency of two times per five days were higher as compared to the frequency of four times per five days.Dietary energy levels had extremely significant influence on serum testosterone concentrations and the number of effective sperms (P< 0.01), and ejaculations and sperm destiny significantly (P< 0.05).The ejaculations, sperm destiny and the number of effective sperms in group Ⅲ were optimum.There was dietary energy×semen collection frequency interaction on serum testosterone concentration (P< 0.01).It could be concluded that the appropriate ME requirement of Shaanbei White cashmere goat bucks was 12.08 MJ/d.

The purpose of this study was to detect the genetic variation of the porcine developmental piuripotency associated 5 (Dppa5) gene and its association with reproductive performance in sow.PCR-RFLP and sequencing methods were used to detect SNPs and analyze its association with reproductive performance in Large White and Landrace sows.The results showed that two missense mutations g.363 T> C and g.844 G> T were detectd in exon of porcine Dppa5.SNP g.363 T> C affected total number born highly significantly in primiparous Large White sow (P< 0.01) and affected litter size and number born alive significantly in Large White and Landrace sows (P< 0.05).SNP g.844 G> T showed a significant impact on total number born in multiparous Landrace and Large White sows (P< 0.05).The results suggested that the SNPs of Dppa5 gene impacted reproductive performance of sows and could be genetic markers.

In order to deeply understand the meat quality and provide the basic information and scientific proof for better heredity selection of rabbit.This experiment option New Zealand rabbit and Californian rabbit were used to construct own DNA pools, respectively for complete sequences amplification of exon 2 and part of intron 1 region of UCP1 gene, two-way sequencing the amplification of the product, BLAST and DNAStar analysis and determine the SNP of rabbit UCP1 gene.The results showed that six SNPs were found in UCP1 gene of the New Zealand rabbit:T135C, C232T, T332C, C360T, T550C and A560G, two synonymous mutation (T135C and C232T) and the other SNPs located in intron region respectively.Using RNA online prediction software forecast UCP1 gene RNA secondary structure prediction software minimum free energy changed from -917.51 kJ/mol to -866.10 kJ/mol for the polymorphisms led to the changes of RNA secondary structure.

The objectives were to research the genetic variations in intron 4 of AQP7 gene in Jiaxian cattle, and analyze the influence of genetic polymorphisms on growth traits.PCR-SSCP technique, DNA sequence analysis and bioinformatics were used to analyze the association between the polymorphisms of the intron 4 of AQP7 gene and growth traits in Jiaxian cattle.The results showed that there were three alleles A, B, C in this locus with the corresponding frequencies of 0.838, 0.076 and 0.086;The polymorphisms of the intron 4 of AQP7 gene had significant association with body weight and chest circumference (P< 0.05), while had no significant association with other growth traits (P> 0.05).The individuals with BC genotype was significantly higher than those of individuals with other genotypes in body weight and chest circumference (P< 0.05).It suggested that BC genotype had positive effect on growth traits, and the correlation was detected between the polymorphisms of the intron 4 of AQP7 gene and growth traits in Jiaxian cattle.So AQP7 gene could be used as a candidate gene for molecular marker assisted selection of growth traits in Jiaxian cattle.

In order to study the value of application in determining the intramuscular fat (IMF) content and other nutrition with dual energy X-ray.Nineteen Mongolian sheep at 8 months of age were chosen to determine the IMF content by soxhlet extractor method and tissue components tester was used to determine the carcass bone density (BMD), fat content of carcass unit minerals (FBMC), the muscle content of carcass unit minerals (LBMC), fat content of the hind legs unit minerals(LFBMC) and hind legs muscle content unit minerals(LLBMC), and these indicators were analyzed by correlation analysis, path analysis and establishing regression equation.The results showed that FBMC had the most significant determinative effects on IMF, and the regression equation consisted of FBMC and IMF was greatly significant differences.Consequently, according to FBMC could predict the IMF in Mongolian sheep carcass.This study provided the necessary basis for the application of dual energy X-ray in the rapid evolution of meat quality.

The aim of the present study was to investigate the characteristics of antimicrobial resistance and prevalence of ESBLs genes of Salmonella isolates from dairy cows in Inner Mongolia.Micro-dilution broth method recommended by CLSI was used to determine the MICs of 22 antimicrobial agents commonly employed in veterinary clinic against the 38 Salmonella isolates.PCR analysis was carried out to detect the presence of 8 ESBLs genes mostly observed in animal origin Salmonella in the isolates with resistance to at least one β-lactam antibiotics, and the features of multi-resistance of the isolates were also analyzed.The results showed that most Salmonella isolates were resistant to trimethoprim(97.4%), sulfamethoxazole(94.7%), and to most of the β-lactam, aminoglycosides, chloramphenicol and tetracycline antibiotics with the resistance rate ranging from 40% to 80%, however, all of the isolates were sensitive to fluoroquinolone drugs.76.3%(n＝29) of the isolates were simultaneously resistant to 6 categories antibiotics and the multi-resistance rate was 94.7%. 85.7%(n＝30) of the β-lactam antibiotics resistant isolates were ESBLs genes positive.Of the six genotype of ESBLs genes found in this study, CTX-M (40.0%) was the mostly encountered ESBLs gene, however, the SHV and PSE types gene were absent in the isolates.A total of fifteen genotypes of the ESBLs genes and nine ESBLs genes combinations were observed in this study, and 16 isolates (45.7%) simultaneously contained two or more than two ESBLs genes.It was indicated that the current situation of the resistance of the Salmonella isolates from dairy cows in Inner Mongolia to the commonly used antimicrobial agents in veterinary clinic was serious, and the prevalence pattern of the ESBLs genes in the isolates was complex, suggesting that the irrational employment of the antibiotic in the treatment of animal infectious diseases was the crucial reason causing the antimicrobial resistance of the isolates.

To investigate the situations of predominant strain and antibiotic resistance of pathogenic Salmonella from chickens in Guangxi Zhuang Autonomous region, Salmonella was pre-enriched and isolated from tissues of clinical suspected chickens, and the Salmonella isolates were identified by biochemical test using ID 32E System for the identification of Enterobacteriaceae of VITEK System ATB Expression, serotypes were determined by slid agglutination test, antibiotic susceptibility was analyzed by ATB VET Susceptibility Test Strip of Enterobacteriaceae antibiotics.The results showed that 34 Salmonella strains were obtained from 310 clinical samples, of which 1 strain belonged to A serogroup, 1 to C2 serogroup, 15 to B serogroup, 14 to D serogroup and 3 to untyped serogroup, and S.typhimurium of B serogroup and S.pullorum of D serogroup were the predominant serotypes.All Salmonella isolates were sensitive to meropenem, imipenem, spectinomycine and apramycine, and the resistance rates to chloramphenicol, kanamycine and gentamicine were less than 10%, and the resistance rates to amoxicillin, streptomycine, flumequine, oxolinique, sulfamethizol, tetracycline and nitrofurantoine ranged between 50% and 90%, while the resistance rates to penicilline, oxacilline, fusidique, rifampicine and metronidazole reached to 100%.The results indicated that the serotype distribution of pathogenic Salmonella from chickens exhibited regional characteristics, and S.typhimurium and S.pullorum were the predominant serotypes in Guangxi Zhuang Autonomous region, and antibiotic resistance of Salmonella isolates was very serious which should be highly concerned.

In order to study the situation of S. aureus small colony variants (SASCVs) and the normal strains hitting the mammary epithelial cells of primary culture, we observed and compared the degree of damage to the cellular structure and the effects on mammary epithelial cells. This test was carried out with a strain of SASCV which was separated by Yunnan Agricultural University Animal Medical Experimental Teaching Center Laboratory via infecting mice mammary epithelial cells in vitro. We put the SASCVs and quality control strains ATCC 25923 respectively into the mammary epithelial cells cultivated in vitro in the laboratory, and observed the morphological change of mammary epithelial cell apoptosis induced by quality control strains ATCC 25923 and SASCVs using ordinary light microscope and scanning electron microscope; At the same time, we also observed the mammary gland epithelial cells apoptosis induced by quality control strains ATCC 25923 and SASCVs using flow cytometry (Annexin V-FITC/PI double staining). The results showed that 3 h after quality control strains and small colony mutant strains infecting the mammary epithelial cells, the mammary epithelial cells could be induced to apoptosis. It showed some typical characteristics of apoptosis such as the nucleus shrivelled, chromatin marginalized, mammary gland epithelial cells could be induced apoptosis, expression was shrinking nucleus, chromatin marginalized, the increase of cell intracytoplasmic vacuoles and so on. After testing using the Annexin V-FITC/PI double staining, we found that compared with the control group, the apoptosis rate of infection group was obviously higher, and the apoptosis rate of the positive control group was obviously higher than that of the experimental group. The results showed that in the process of infecting cells, the SASCVs reached a certain number and then stopped; The apoptosis rate of SASCVs was obviously lower than the normal of S.aureus. Therefore, SASCVs could also induce mammary epithelial cells apoptosis. It had the typical morphological characteristics of apoptosis. It provided experimental basis for the prevention and control of SASCVs chronic mastitis cows.

To investigate the pathogenicity of A/Hero/Guangdong/C1/2013(H5N6), an AIV strain isolated from Heron on ducks and mice, pathogenicity of this new virus strain and changing of histopathology as well as a preliminary study on its biological characteristics were studied by virus challenges test via intranasal and eye-drop and in chicken via jugular vein injection.Results showed that the EID₅₀ of this strain of virus was 10^-8.16/0.1 mL in embryonated chicken eggs and the intravenous inoculation of pathogenic index (IVPI) was 2.76.In ducks and mice, the 50% lethal doses (LD₅₀) of it were determined to be 10^-4.0/0.2 mL and 10^-4.67/0.05 mL, respectively.Symptoms of infection included loss of appetite, depression, swollen head and tears after being infected with 10⁶ EID₅₀ per duck by intranasal and eye-drop administration.Most of ducks died 4 to 7 days post-infection, liver, lung and kidney still eliminated toxicants at day 7 post-infection.Anatomy showed symptoms of pericardial effusion, pulmonary congestion and kidney enlargement, while pathological section showed pathological change like karyopycnosis and inflammatory cell infiltration in heart, liver, kidney and spleen.Mice developed symptoms of infection like loss of appetite, depression, shaggy coat and ruffled coat after being infected with 5×10⁵ EID₅₀ per mouse by intranasal and eye-drop administration.Most of the infected mice died 5 to 7 days post-infection and only liver still eliminated toxicants at day 7 post-infection.Although organ anatomy showed no obvious pathological changes, pathological section showed pathological changes like karyopycnosis and inflammatory cell infiltration in heart, kidney and lung.Our research demonstrated that this H5N6 subtype AIV had a strong pathogenicity and could be defined as a highly pathogenic AIV strain as its IVPI was greater than 1.2.Our work laid a theory foundation for study, prevention and control of H5N6 subtype AIV.

The 90 chickens were randomly divided into control group, E.necatrix one time infection group and E.necatrix two times infection group.We used duplex PCR method to test the expression changes of HSP-90 mRNA in intestinal tissue of chickens infected with E.necatrix and researched the effects of it comprehensively and systematically.The results were showed that after 0 to 14 days the expression of HSP-90 mRNA in intestinal tissues of 14-day-old chickens infected with E.necatrix presented a upward trend though it was lower than the control chickens.These chickens were infected with E.necatrix again at 28 days, HSP-90 mRNA expressions of duodenum, jejunum and ileum decreased and then increased.However, it always showed a downward trend in appendix.When 28 days chickens were infected with high-does E.necatrix once, HSP-90 mRNA expressions of duodenum, jejunum and appendix were in a declining curve.While it was increased originally and then decreased in ileum.This study indicated that HSP-90 mRNA expressions in intestinal tissue of chickens infected with E.necatrix were inhibited at first, and the expression quantity was closely related to the degrees of intestinal tissue damage.

In order to compare the resistance of E.coli from pigs of different regions under same breeding mode in Xinjiang, we collected fecal samples from pig farms of Hutubi (207), Manasi (210) and Changji (210) in a certain scale, respectively, a total of 627 samples, and isolation rate of E.coli from fecal samples were all 100.0%.The broth dilution method was used to detect resistance of E.coli to antimicrobials, and we compared the differences of resistance rate of E.coli from pig farms in three regions by chi square test. E.coli from pig farms of Hutubi region to apramycin, amikacin, amoxicillin-clavulanic acid and ceftiofur existed extremely significant differences comparing with another two regions (P< 0.01).E.coli from pig farms of Manasi region to ciprofloxacin existed significant differences comparing with another two regions (P< 0.05), and existed extremely significant differences to norfloxacin (P< 0.01).E.coli from pig farms of Changji region to norfloxacin had extremely significant differences comparing with Hutubi region (P< 0.01), and existed significant differences to amoxicillin-clavulanic acid and ampicillin comparing with Manasi region (P< 0.05).3 to 8 resistant E.coli strains accounted for 94.7% in Hutubi region, 3 to 7 resistant E.coli strains accounted for 89.5% in Manasi region and 4 to 8 resistant E.coli strains accounted for 82.4% in Changji region, respectively.There was no significant difference among different regions on multi-drug resistance (P> 0.05).The results indicated that E.coli from pigs of different regions under the same breeding mode had different characteristics of resistance.In addition, the resistance of E.coli from pigs were very serious and mainly multi-drug resistant, drug resistance patterns were diversified.

This study used Real-time PCR to detect the changes of expression quantity of claudin 3 (CLDN-3), cytokeratin 8 (KRT8) and mucin 1 (MUC1) three proteins mRNA in IPEC-J2 cells at three different time points.IPEC-J2 cells were infected by TGEV, we explored whether Lactobacillus amylovorus metabolites could enhance cell barrier function by maintaining or improving mRNA expression of CLDN-3, KRT8 and MUC1, whether had effects on the barrier function of IPEC-J2 infected by TGEV.The results showed that mRNA expressions of MUC1, CLDN-3 and KRT8 in the normal cell control group did not change significantly with time (P> 0.05); MRS medium culture group did not change significantly compared to the normal cell control group(P> 0.05); Gene expressions of MUC1, CLDN-3 and KRT8 proteins in Lactobacillus amylovorus metabolites alone treatment group showed significant time-dependent effect (P< 0.05).After being treated 24, 36 and 48 h, gene expressions of MUC1, CLDN-3 and KRT8 proteins in Lactobacillus amylovorus metabolites+TGEV experiment group was significantly higher than virus control group (P< 0.05).Lactobacillus amylovorus metabolites could improve MUC1, CLDN-3 and KRT8 expressions in IPEC-J2 cells, while being able to negatively regulate MUC1, CLDN-3 and KRT8 expressions that TGEV induced in IPEC-J2 cells, thereby inhencing the barrier function of IPEC-J2 cell against TGEV.

In order to investigate coccidium infection of a Rex rabbit farm in Fujian province, 494 fecal samples were collected from different ages of Rex rabbits and examined by McMaster's method and the method of saturated saline flotation.The results showed that the total prevalence was 87.45% and the mean oocysts per gram (OPG) value was 2.56×10⁴.The prevalences of coccidia among each rabbit age were different, and it was highest at the age of about 40 d (98.61%) and about 65 d (98.95%) rabbits.The prevalence and OPG values at the age under 30 d (19.23% and 331) were significantly lower, and the distribution pattern of OPG values was negative binomial distribution in rabbits by using fitting of distributions.In total, 11 species of Eimeria were identified from oocyst-positive samples.The infection of Eimeria was mixed infection with more than two Eimeria species, and mostly of which were infected by 3 to 7 Eimeria species.The dominant species of Eimeria were E.perforans, E.media, E.exigua and E.magna, and their prevalences were 20.15%, 16.79%, 13.94% and 13.48%, respectively.

The Animal Nutrition is an important professional foundation course of animal science in agricultural and forestry universities.At present, the animal husbandry industry shifts from traditional agricultural herdsman cage-free to large farms breeding mode of modernization, intensification, industrialization, industrialization, high-efficiency production and securitization.The traditional single professionals were relative surplus, but composite professional talents with innovation ability were relatively scarce.The author has put forward the practical teaching reform train in view of teaching reform of Animal Nutrition based on cultivating innovative talents as the goal, and provided some reference to improve the teaching quality and effectiveness of the course of Animal Nutrition.