The purpose of this study was to clone the full-length cDNA of tree shrew's Tssk6 gene, to elucidate its tissue expression profiles and fundamental bioinformatical characteristics. In the present study, the full-length cDNA of tree shrew's Tssk6 gene was cloned with specific primers on the basis of predicted mRNA sequence of tree shrew's Tssk6 gene in GenBank, the mRNA expression of Tssk6 gene in various tree shrew's tissues was investigated via Real-time PCR. In addition, the bioinformatics on nucleotide sequence of CDS region of Tssk6 cDNA in tree shrew as well as putative protein structure had been analyzed. The results showed that:The CDS region of tree shrew's Tssk6 cDNA contained 822 bp encoding 273 amino acids; Tssk6 gene specifically expressed in tree shrew's testis; Based on the molecular phylogenetic tree constructed with CDS of Tssk6 cDNA among tree shrew and other eight mammalian species, it was inferred that tree shrew was genetically close to primates like human, chimpanzee and rhesus monkey, while it was genetically far to rodents like mouse, rat and golden hamster. This study would lay a foundation for the further study of Tssk6 gene function in tree shrew.

The objective of this study was to clone the acyl-coenzyme A oxidase 1 (ACOX1) gene of Luchuan pig and analyze its genetic structure with bioinformatics. A pair of special primer was designed according to predicted sequence of pig ACOX1 gene in GenBank. The ACOX1 gene was amplified by RT-PCR, the physicochemical property and secondary structure of ACOX1 gene were systemically analyzed by bioinformatics techniques. The results showed that ACOX1 gene fragment included an 1 986 bp whole length CDS (coding 661 bp amino acids). The sequence multi-aligned results showed that Luchuan pig shared 99.5%, 85.5%,87.1%,66.3%,75.6%,84.0%,82.3%,81.1% and 69.1% of similar nucleotide sequence with that of pig,human, zebrafish, chicken, rhesus monkeys, mice, rat and frog, respectively. The prediction of ACOX1 secondary structure showed that Luchuan pig ACOX1 had no signal peptide and transmembrane structure. This study suggested that the whole CDS sequence of ACOX1 gene was successfully cloned in Luchuan pig, and the cloning and analysis of ACOX1 gene provided an important foundation for further studying on the fatty deposition and lipornetabolism of ACOX1 gene in Luchuan pig.

In order to obtain the full-length cDNA of sika β-defensin-1 (siBD-1),5'-RACE and 3'-RACE primers were designed according to the partial sequences of siBD-1 that obtained from this study group, cDNA ends (RACE) technology were used to amplify siBD-1 cDNA from the total RNA of tongue mucosa tissue in sika, and amplified products were cloned into pMD18-T vector and subjected to PCR, restriction endonuclease digestion and sequencing. The results indicated that the length of siBD-1 cDNA 5'- and 3'- end were about 172 and 299 bp. The full-length siBD-1 cDNA was 418 bp (GenBank accession No. HM588696.1) and includes an 5'-untranslated region (UTR) of 89 bp, a open reading frame (ORF) of 192 bp, a stop codon of TAA, 3'-UTR of 118 bp and Ploy(A)16. The sequence homology showed that siBD-1 shared the greatest identity (90.6%) with buffalo enteric β-defensin, the higher identity with cattle (EBD, LAP, TAP, BNBD-4), goats (GBD-1, GBD-2), reindeer (reBD-1), sheep (sBD-1, sBD-2) and camel (caBD-1), that were 83.2%, 83.1%, 87.3%, 87.0%, 87.5%, 87.5%, 84.4%, 79.9%, 77.1% and 70.5%,respectively,and correspondingly low identity were 60.3% and 72.4% with horse (hoBD-1), pig(pBD-1), the lowest was 16.0% with human (hBD-2). The mature peptide was consisted of 38 amino acids, and contained 9 positively charged residues.

This study was aimed to construct the dual luciferase reporter vector which contained KLF3 gene 3'-UTR and its mutation vector, and analyze the possible miRNAs which could regulate KLF3 gene expression. In this study,The KLF3 gene 3'-UTR was amplified by polymerase chain reaction (PCR) and inserted in the dual luciferase reporter vector which was digested by Xho Ⅰ and Not Ⅰ;With Targetscan to predict the miRNAs which might interact with KLF3 gene 3'-UTR;293T cells was treated with recombinant vector or its mutation vector or empty vector and miRNA mimics or control by Lipofectamine 2000.The dual luciferase reporter assay system was used to compare the activity of luciferase. The results showed that KLF3 gene 3'-UTR might be the action points of miR-21;Double fluorescent report results showed that miR-21 mimics group (0.6900±0.0144) was decreased 31% than the mutation vector group (1.000±0.0688) and the blank control group (1.000±0.0159) with the luciferase activity of 293T cells. This study successfully constructed KLF3 gene 3'-UTR dual luciferase reporter vector and its mutation vector;Preliminary evidence showed that miR-21 could regulate the expression of KLF3 gene.

The aim of this study was to determine the optimal method of the yeast cell wall breaking which could obtain the maximum extraction rate of yeast polysaccharide. The extraction rate of yeast polysaccharide and the cell wall broken rate were used as the evaluating indexes. The conditions of ultrasonic disruption, repeated freezing and thawing, ultrasonic cleaning and protease treatment were optimized to select the appropriate parameters in cell wall breaking. The effects of these 4 methods and the synergistic action of ultrasonic disruption and protease treatment were compared. The results showed that when the ultrasonic cleaning time was 40 min and ultrasonic disruption time was 50 min, the water content of slurry was 15% and the times of freezing and thawing was 3, and the protease treatment was carried out at 40℃ for 15 h with enzyme concentration 300 IU/g and pH 5.0, respectively, the extraction rates of yeast polysaccharides were the highest. The effect of protease treatment was the best among these 4 cell wall breaking methods, its extraction rate of polysaccharide was 149.81 mg/g and the cell wall broken rate was 53.49%. The effect of combined way was better than that of single treatment, but the synergistic action would destroy the integrity of polysaccharide's structure. So the protease treatment was thought an ideal cell wall breaking method of yeast.

There are abundant microorganisms in the gastrointestinal tract of chicken, which play important roles in the metabolism and immunity of the host. The development of high-throughput sequencing technology for intestinal microbial research provides a new way of thinking. The article elaborates that metagenomics and effects of changes in intestinal microbial structure on the host and different feeding conditions, host physiological characteristics effect on intestinal microbial. This article focuses on the analysis of correlation between the intestinal microbial colony structure and characteristics of nutrient metabolism, immune function, and healthy growth of chicken on the level of metagenomics, and we hope to provide ideas for the study of intestinal microbial.

To seek out proteins interact with VP2 structural protein of porcine parvovirus (PPV) in ST cell and study the molecular mechanisms of viral infection and pathogenicity, a ST cDNA yeast hybrid library was created. Firtly,the total RNA of ST was extracted using RNeasy Min Kit,and double strands cDNA was synthesized by SMART technique. Then, the ST cDNA library was successfully created by homologous recombination in yeast, the titer was 8.1×10⁸ CFU/mL, the inserts varied from 250 to 1 000 bp,the average of inserted fragments was about 500 bp,and the recombination rate was 96%.These datas indicated that the yeast two-hybrid cDNA library was successfully constructed and might be useful for screening the host proteins interacting with structure protein of PPV.

To determine the roles of miRNA in the developmental stages of Xiang pig,we investigated the miRNA expression patterns in different months old of Xiang pigs.The total RNA from 3 and 7 months old of Xiang pigs were collected separately. Then,the total RNA were hybridized with the miRNA microarray and the threshold (fold-change>2, P<0.05) were used to identify the different expression miRNA. The quantitative Real-time PCR was performed to verify the microarray result. Compared to the 3 months old group,there were 11 miRNA down regulated and 9 miRNA were up regulated in the 7 months old group. The lowest down regulated miRNA was miR-27b whereas the highest up regulated one was miR-155.The result of quantitative Real-time PCR consistented with the microarray result. The result suggested that some of the dysregulated expression of miRNA might participate in the lipid metabolism in hepatic cells.

The purpose of this study was to observe and analyze the ability of biofilm formation of the Enterococcus faecalis (E.faecalis) XJ05 and to detect if the E.faecalis which could induce lamb encephalitis carried the biofilm-associated genes. The E.faecalis XJ05 was used as the sample. The biofilm model was established by the 96-well microtiter plate and read by microplate reader in the D_{595 nm} at the same time observed by the inverted microscope. The primers were designed to examine if the 11 strains of E.faecalis carried the biofilm related genes and then the amplified products were sequenced and analyzed for homology. The results showed that the amount of BF produce in XJ05 strain was significantly different from blank control group at different time (P<0.05), and the highest D_{595 nm} value was at 24 h and significant difference compared with other time (P<0.05). The biofilm of 6 h was scattered when observed under the microscope and the time between 12 to 24 h biofilm to be more denser, after 24 h the biofilm became to degradate and loose gradually. It turned out that the 11 strains of bacteria carried different biofilm-associated genes, 8 biofilm-associated genes were found in 6 strains of bacteria, 7 biofilm-associated genes were find in 1 strain of bacterium, 6 biofilm-associated genes were found in 1 strain of bacterium, 2 biofilm-associated genes were found in 1 strain of bacterium, 2 strains of bacteria could not text out anyone of the detected genes.The homology of all detected genes were all between 93.3% to 100.0%. It concluded that the E.faecalis XJ05 could form the completed biofilm and there were 9 kinds carried the biofilm-associated genes in 11 strains of E.faecalis.

E.coli K1 strain is a representative strain of neonatal sepsis and meningitis, which causes disease by blood circulation to the brain. The molecular mechanism of E.coli K1 that adhering and invading the brain microvascular endothelial cells (BMECs) and cross the blood brain barrier (BBB) has been focused on by many scholars. In this review, we focused on the gene regulation mechanisms and signaling pathway of E.coli K1 crossing the blood brain barrier to understand the molecular mechanism of the infection caused by E.coli and provide reference for prevention and treatment programs.

This study was aimed to explore the developmental change of lipid droplet in liver of chicken embryos.120 fertilized eggs were incubated, and 6 embryos were selected randomly every day and their livers were removed for measuring the formation and development of lipid droplets from E7 to E21 during the incubation.The methods of frozen section and Sudan Ⅲ staining were used. The results showed that:①Lipid droplets emerged early on the 9th day of chicken embryos incubation (E9). The quantities and sizes of lipid droplets became more and larger with the increasing of chicken embryos age. ②Quantities of lipid droplets in every hepatic lobule decreased from central veins to the margin of hepatic lobule along hepatic cords before E18, while more lipid droplets distributed at the margin of the liver than that in the center of liver after E18. ③The quantities and sizes of hepatic lipid droplets were significant difference during the three stage of E9-E14, E15-E17 and E18-E21 (P<0.05). The average sizes of hepatic lipid droplets were 3.2, 5.7 and 10.8 μm, respectively, and their average numbers were 67,327 and 397 n/mm², respectively. In conclusion, the quantities and sizes of hepatic lipid droplets became more and larger with the increasing of chicken embryos age.

This research adopted fermentation gas production method in vitro to evaluate the associative effect of whole corn silage (WCS) and millet straw (MS).The WCS was mixed with MS in the proportion of 100:0,80:20,60:40,50:50, 40:60,20:80 and 0:100 with 3 replicates per group.The gas production,pH,ammonia nitrogen (NH₃-N), volatile fatty acids (VFA), microbial crude protein (MCP) and dry matter loss (DML) for 24,48 and 72 h of varying forage combinations were analyzed.Single factor associative effects index (SFAEI) and multiply factors associative effects index (MFAEI) of different combinations were calculated. The results showed as follows:There were significant differences of gas production among different combinations (P<0.05),and the best combination was the WCS80 group. With creasing of the content of MS,DML decreased,but the DML of WCS80 and WCS100 were similar (P>0.05). The MCP concentration of WCS100 and WCS80 was extremely significantly or significantly higher than other groups (P<0.05;P<0.01). The VFA concentration showed significant differences among different combinations (P<0.05).Among the different combinations of WCS and MS,the MFAEI of WCS80 was the largest,and that were 0.5517,0.5094 and 0.9860 at 24,48 and 72 h,respectively. The best combination of WCS and MS was 80:20.

In order to define the quality characteristics of different anatomical locations of Yunling cattle,the pressing loss rate,cooking loss rate,shear force,pH, L^* value,a^* value,b^* value,crude protein content,crude fat content and moisture of supraspinatus (SU),infraspinatus (IF),triceps brachii (TB),rhomboideus (RH),serratus ventraliscervicis (SVC),splenius (SP),psoas major (PM),longissimus dorsi (LD),latissimus dorsi (LA),semimembranosus (SM),semitendinosus (ST),biceps femoris (BF),gluteus medius (GM),rectusfemoris (RF),vastus lateralis (VL), tensor fasciae latae (TFL) from Yunling cattle carcass were determined.The quality traits of Yunling cattle meat among different anatomical locations were analyzed using variance,correlation and standardization analysis. The result showed that there were significant differences in quality traits among various anatomical locations (P<0.05). The SU,IF,PM,SVC and TFL had higher tenderness comparing to other muscles with shear force values less than 4 kg,while the water holding capacity of SP,ST and VL was poorer than other muscles. The ST had the highest L^* value among all muscles. There was significantly positive correlation between shear force and pressing loss, cooking loss rate. The LD and ST had similar quality characteristic,while BF,GM and VL had similar quality characteristic. It was showed that different parts of muscle had significant effect on Yunling cattle meat quality.The quality of forequarter meat was better, which could be used to develop high-end products.

To study the effects of N-hydroxymethyl methionine calcium (N-HMM-Ca) supplementation levels on serum nitrogenous compounds and free amino acids of lactating dairy cows. 30 lactating Holstein dairy cows with similar parities,milk yield and days in milk were randomly assigned into 5 groups. The control group was fed with basal diet,test groups were fed with basal diet supplemented with 0.15%,0.30%,0.75% and 1.50% N-HMM-Ca,respectively. The experiment was lasted for 56 d,including 14 d of adaptation period and 42 d of experimental period. The results showed as follows:①Comparing with control group,the total protein,albumin and globulin in experimental groups were not significantly affected (P>0.05).On the 14th day,serum creatinine concentration in 1.5% level group was significantly lower than control group (P<0.05);On the 42th day,urea nitrogen concentration in 1.5% level group was significantly lower than control group (P<0.05),while the concentration of uric acid in 0.75% level group was extremely significantly higher than that in other groups (P<0.01).②On the 14th day,the concentration of serum Glu,Ser,Arg,Leu,Ile,Lys and total free amino acids in 0.15% level group was highest,which were significantly or extremely significantly higher than control group (P<0.05;P<0.01).On the 42th day,serum Arg,Cys and the total free amino acids in 0.15% level group were the highest. And serum Glu,Ser,Tyr,Met,Val and Lys in 0.30% level group were highest,which were significantly or extremely significantly higher than the control group (P<0.05;P<0.01). In summary, adding 0.15% or 0.30% N-HMM-Ca on diet of lactating dairy had no significant effect on serum nitrogen content (P>0.05),but adding 0.15% N-HMM-Ca could increase the concentration of serum free amino acids. Considering all the facts,the recommended amount of N-HMM-Ca on diet of lactating dairy cows was 0.15%.

The objective of this study was to evaluate of the growth performance, serum biochemical indices, liver antioxidant parameters and ability of anti-infection with Aeromonas hydrophila by adding the fermented Chinese medicine by Bacillus Subtilis to the carp fish feed, and to find out the appropriate supplemental level of fermented Chinese medicine in diets. Carp fingerling with an initial body weight of (35±0.5) g were set up to 6 experimental diets supplemented with 0 (control group), 0.1‰, 0.5‰, 1‰, 2‰ and 3‰ fermented Chinese medicine for 56 days, respectively. Each group had 30 fish and set up 3 repetitions. After the feeding trail,the fish were challenged with Aeromonas hydrophila to calculate the relative percentage survival (RPS).The results showed as follows:Supplementing fermented traditional Chinese medicine by Bacillus subtilis, the weight gain rate (WGR), specific growth rate (SGR), condition factor (CF), the activities of lysozyme (LSZ), complement 3 (C3),total protein (TP),total anti-oxidation capacity (T-AOC),total superoxide dismutase (T-SOD) and catalase (CAT) in serum of common carp were enhanced at different degrees, while the feed coefficient (FCR)and malondialdehyde (MDA) in serum were declined at different degrees.When the fermented Chinese medicine supplemental levels were 2‰ and 3‰,the effects were significant (P<0.05)(except for C3, ALP and Cr).The experimental results of the virulence of common carp infected with pathogen Aeromonas hydrophila showed that,the fermented herbal could obviously improve the ability of anti-infection, the immune protective rate of the group with 2‰, 3‰ doses of the fermented Chinese medicine could reach 77.23% and 74.63%, respectively. From what had been discussed above, the group with 2‰ and 3‰ doses of the fermented Chinese medicine had significant effect on the growth performance, serum biochemical indices, liver antioxidant parameters and Aeromonas hydrophila anti-infection ability. Considering the cost of adding fermented Chinese medicine, the recommendations for the best additive quantity was 2‰.

This research was to study the effect of bacterial lipopolysaccharide (LPX) and dexamethasone (DEX) on the growth performance,nutrient metabolism and blood biochemical indexes and intestinal morphology development of broiler chickens. 224 AA broilers with similar body weight and 1-day-old were chosen and divided into 4 groups randomly,which were immune group (NV group),normal immune group (CV group),immunosuppression group (DEX group) and immune hyperthyroidism group (LPS group) with 7 replicates per group and 8 broilers per replicate. At 15,17 and 19 d,chicken of LPS and DEX groups were intramuscular injected with 500 μg/(kg·BW) LPS or 2.5 mg/(kg·BW) DEX,respectively,while chicken of NV and CV groups was injected with normal saline in the same volume. The experiment was lasted for 42 d with 7 d preliminary trial period. The results showed that the average daily feed intake (ADFI) and average daily gain (ADG) were decreased significantly (P<0.05) by LPS and DEX immunological stress,there were was no significant difference in feed to gain ratio (F/G) among experimental groups (P>0.05).The apparent metabolic rate of organic matter (DM) was no significantly affected by LPS and DEX immunological stress,while that of dry matter (OM),crude protein (CP),Ca and P were declined significantly at 36-42 d (P<0.05). At 21 and 42 d,GLU,UA and MDA levels were not significantly different among experimental groups (P>0.05).At 21 d,the contents of serum IL-6 and TNF-α in DEX and LPS groups were significantly higher than those in NV and CV groups (P<0.05),but there was no significant difference at 42 d (P>0.05).The wall thickness,mucosa thickness, villus height and the ratio of villus length to crypt depth were reduced by LPS and DEX at 21 and 42 d of broilers. In summary,LPS and DEX had an important influence on the apparent digestibility of DM,CP,Ca and P.DEX and LPS could damage the intestinal morphology of broilers,reduce the apparent metabolic rate,affect the feed intake and body weight,decrease growth performance.

This experiment was conducted to study the impact of adding linseed oil and malic acid on rumen fatty acid metabolism and feed degradation rate in Yanbian Yellow cattle. 4 Yanbian Yellow cattle weighted (350±15) kg, and fitted with permanent rumen fistula were used as 4×4 Latin square design. The cattle in control group (CON group) were fed basal diet, in LO group were fed the basal diet supplemented with 475 g/d linseed oil, in LOM group were fed the basal diet supplemented with 475 g/d linseed oil and 160 g/d malic acid, in LOMS group were fed the basal diet supplemented with 475 g/d sesame oil, 160 g/d malic acid and 47.5 g/d NaHCO₃. In the third and fourth day during the test period, rumen fluid was collected and measured for fatty acids. In the fifth and sixth day, nylon bag method was adopted to determine the feed degradation rate in the rumen. The results showed that the contents of the rumen fatty acids in three experimental groups were significantly higher than those in the CON group (P<0.05), excepted the contents of t11-C18:1 at 6 h. Compared with CON group, the rumen degradation rates of DM, OM, CP and NDF in LO group were all reduced. Compared with LO group, degradation rates of DM, OM, CP and NDF in LOM and LOMS groups were improved in a certain extent. The results showed that dietary linseed oil and malic acid would increase the rumen fluid fatty acids in Yanbian Yellow cattle. Linseed Oil had a trend to decline the feed degradation rate in the rumen, but the trend was opposite when combined with malic acid.

Fatty acids are thought to supply energy for cell traditionally. Recent studies have shown the effect of fatty acids on cellular growth, development and metabolism. Skeletal muscle is an important place for fatty acids consumption, and it's also the main target where fatty acids play its role. Palmitic acid is closely related to skeletal muscle insulin resistance (IR); Oleic acid can increase skeletal muscle fatty acids oxidation and triglyceride synthesis through peroxisome proliferator activated receptor;Linolenic acid relieves skeletal muscle insulin resistance by increasing glucose utilization; Conjugated linoleic acid (CLA) influences skeletal muscle metabolism via multiple signal passways;Docosahexaenoic acid (DHA) and arachidonic acid (ARA) can significantly change myosine heavy chain gene expression;Eicosapntemacnioc acid (EPA) has some effect on skeletal muscle glucose metabolism,mitochondrial action and Apelin system.This review focuses on the effects of different fatty acids on skeletal muscle.And we will have a better understanding about the diverse regulatory mechanism of various fatty acids, and we also hope that this paper will provide some theoretical foundations for the research and development of fatty acids products.

Animal transport has always been an important part of the production of animal husbandry.But the transport stress disorder caused by long distance transportation has become a serious problem which is harmful to the development of beef industry. This paper reviewed the effects of transport stress on animal behavior,blood biochemical indexes, immune function and meat quality. It had important practical significance for future research related to transport stress mechanism and promoting the development of animal husbandry in China.

To explore the changes in body size indexes,carcass traits and meat quality of Baixi pig×Turok hybrids F1 (DBF1),16 Baixi pigs and 6 DBF1 pigs which were health and 10 months old were chosen for determining the body size indexes,slaughter performance and meat quality. The results showed that there was no significant difference of body size indexes between Baixi pig and DBF1 pigs (P>0.05).The eye muscle area (28.33 cm²),meat color (3.00),marbling (3.50) of DBF1 pigs kept a good level,while the unsaturated fatty acid content (64.33%),the intramuscular fat content (4.08%) and leg to buttock ratio (31.32%) were increased, back fat thickness (24.69%),total flavor amino acid amount (48.46%) were significantly or extremely significantly decreased (P<0.05;P<0.01),and the muscle tenderness,water retention capacity were significantly reduced (P<0.05). In conclusion,the DBF1 pigs could maintain the good meat performance,sensory index and tender degree of muscle,improve the hindquarters meat performance,lean meat and the intramuscular fat contents,but the muscle tenderness,nutritional value, freshness and water retention capacity were declined.

Proline is a cyclic α-imino acids,meanwhile it also can be considered as a functional amino acid for mammals and plays an important role in protein synthesis,organism metabolism,wound healing,anti-oxidant and immune reactions. The studys have found that extra dietary supplementation of moderated proline has great significance in improving the growth and development of conceptus,piglets, embryo,mammary gland and placenta of sows,at the same time it also can prevent intrauterine growth restriction. The author reviewed structure characters and physicochemical properties,anabolism and catabolism of proline,and especially focused on its effects on the reproductive performance of pregnancy sows and potential regulation mechanism aiming to provide some help for the researchers in the field of sows.

In order to assess the purity of Luchuan pig populations, four South Chinese local pig breeds including Putian Black pig, Yuedong Black pig, Dahuabai pig, Bama miniature pig and three foreign pig breeds including Duroc pig, Yorkshire pig and Landrace pig were studied as controls by sequencing of PCR products, MC1R and KIT genotypes in 56 Luchuan pigs were analyzed in this study. Sequencing results indicated that a splicing mutation (G>A) was presented in the first base in intron 17 of KIT gene in both Yorkshire pig and Landrace pig, in contrast, the wildtype GG of KIT gene was presented in Luchuan pig, four south Chinese local pig breeds and Duroc pig.Compared with Hainan wild boar, South Chinese local pig breeds had two missense mutations 95Val > Met and 102Leu > Pro in the coding region of MC1R gene;Compared with Yorkshire pig, Landrace pig and Duroc pig, South Chinese local pig breeds had 5 to 6 SNPs in MC1R gene 5'UTR, and in addtion, an A base deletion in MC1R gene 3'UTR. Furthermore, we found one litter of Luchuan pig with abnormal coat color.The results showed that the presentation of two distinct MC1R genotypes E^D1 and E^p in both litters and the sow,but only E^D1 in the boar. Considering E^p was derived from Pietrain pig, we preliminarily considered that the genome of the sow might be infiltrated with foreign pig breeds. In summary, we detected the genotypes of the coat color genes KIT and MC1R in eight pig breeds, confirmed the molecular differences of coat color between Chinese local pig breeds and foreign pig breeds, which could be useful for the further investigation of the molecular mechanism of pig coat color.

The purpose of this study was to research the sequence characteristics and the expression of PLIN2 gene, and to analyze the relationship between the expression and the ovaries in different estrus periods. According to the gene sequence of bovine in GenBank, the primer was designed. The PLIN2 gene of yak was cloned by RT-PCR, and the expression quantity of the PLIN2 gene in different tissues of yak was detected by the semiquantitative RT-PCR. Meanwhile,the differential expression of the PLIN2 gene in different ovarian estrus periods of yak was detected by quantitative Real-time PCR. The results showed that the length of the PLIN2 gene was 1 353 bp in CDS, encoding 450 amino acids. The PLIN2 protein was predicted to be non-seretory and lacking cross-membrane domain. It was 99.4% homology identity to that of corresponding cDNA from bovine, which showed that the PLIN2 gene was quite conservative in the process of evolution. The expession profile of PLIN2 was wide in yak tissues, but there were some differences in different tissues. The expression quality was higher in kidney, breast and ovary, while it was almost nothing in lung. The result of quantitative Real-time PCR indicated that yak PLIN2 gene was expressed in different ovarian estrus periods, and the highest expression during the luteal phase, which indicated the PLIN2 gene played an essential role in yak estrous cycles.

In order to understand the epidemiology and evolution of PCV-2 in Jiangxi province, a pair of primers was designed according to the PCV-2 gene sequence published in GenBank. After PCR amplification, we got the whole genome sequence of 9 strains PCV-2 isolates, and the nucleotide and protein sequences were analyzed, the genetic evolutionary tree was constructed. The results showed that the complete genome of 8 out of the 9 strains were 1 767 bp in length and one strain was 1 768 bp. By analyzing the whole genome sequences of the nucleotide,the homology of nucleotide sequences of the 9 strains was 94.7% to 99.9%.Compared with other whole genome sequences of PCV-2 in GenBank, the homology was 94.3% to 99.8%. The homology of nucleotide sequences of the ORF1 of the 9 strain was 96.9% to 100.0%. ORF2 and ORF3 encoding protein amino acid sequence had some locus mutation. Phylogenetic tree analysis showed that the 9 strains could be divided into 3 genotypes,5 strains belonged to PCV-2b, 3 strains belonged to PCV-2d, and 1 strain belonged to PCV-2a. This study was helpful to monitor and control of PCV-2 in Jiangxi province.

Follicle-stimulating hormone (FSH), a glycoprotein secreted from pituitary, can regulate ovarian function through the FSH receptor (FSHR).To evaluate the effects of FSHR gene on the litter size of Xiang pig population, the polymorphisms of FSHR gene exon 10 were detected by polymerase chain reaction (PCR) and sequencing methods, and the effect of polymorphisms in FSHR gene exon 10 on litter size traits in Xiang pigs was analyzed using the least square analysis.In eight candidate sites, there were two polymorphic loci were detected out from Xiang pig population, which were rs322800083 (C276T) and rs332115220 (T601C). Moreover, a new SNP novel 1 (G-92A) was discovered.Four haplotypes were constructed between rs322800083 and rs332115220 sites, in which C-T was the main haplotype (45.0%) and presented closely linkage disequilibrium (r²=0.334) in Xiang pig population. Furthermore, the haplotype C-T corresponded to the highest total number born (TNB) at the second fetus of Xiang pigs, haplotype C-C to the lowest TNB. It was notified that haplotype C-T produced 0.99 TNB of Xiang pigs more than haplotype C-C (P<0.05). And the TNB at the third fetus was also the highest in haplotype C-T, which produced 1.61 and 2.33 TNB more than haplotypes T-C and C-C (P<0.01). The genotypes of locus rs332115220 was weakly correlated with the third fetus of Xiang pig population (ρ=0.429).The results suggested that haplotype C-T could be used as a molecular marker to assist selection for the litter size of Xiang pigs.

This study was designed to compare the efficiency of porcine oocytes vitrified at the GV and MⅡ stages. The vitrified GV oocytes were matured in vitro and then evaluated their cortical granule distribution with immunofluorescence, CD9 protein level with Western blotting and zona pellucida dissolution time with pronse digestion method, as compared to vitrified MⅡ oocytes.The results showed that the survival percentages of oocytes vitrified at the GV stage were significantly lower than those vitrified at the MⅡ stage after 2 h of warming (P<0.05);However, there was no difference in maturation rate between vitrified and fresh oocytes (P>0.05). The oocytes vitrified at MⅡ stage resulted in significantly decreased normal cortical granule distribution and CD9 protein level (P<0.05), but no such results were found in vitrified GV oocytes after maturation (P>0.05). In addition, vitrification did not affect the dissolution time of zona pellucida in GV and MⅡ oocytes (P>0.05). The data demonstrated that despite of lower survival rate compared with MⅡ oocytes, vitrified porcine GV oocytes after maturation showed normal polar body extrusion, cortical granule distribution and CD9 protein level.

Single nucleotide polymorphisms (SNPs) in exon 2 of insulin-like growth factor Ⅰ receptor (IGF-ⅠR) gene were detected by PCR-RFLP in Bian chicken,which was to find the correlation markers of growth and reproduction traits in Bian chicken.The results showed that AluⅠsite displayed polymorphic, and a G→A mutation at the position of the 376 bp in exon 2 was occurred. There were three genotypes (GG, GA and AA), the genotypes frequencies were 0.687, 0.296 and 0.017, respectively, GG genotype was the dominant genotype, the frequency of G allele was 0.835, which was the dominant allele. This site was in Hardy-Weinberg equilibrium (P>0.05).Correlation analysis showed that GG genotype had significantly higher body weight than those of GA genotype at the age of 8, 14, 16 and 18 weeks (P<0.05);The GG geno type had extremely significantly higher body weight at first egg than those of GA genotype (P<0.01),but it didn't mean that this locus could be used as a genetic marker for reproduction traits in Bian chicken. This results suggested that the AluⅠsite (G→A) in exon 2 of IGF-ⅠR gene could be used as a genetic marker for growth traits in Bian chicken.

Cryopreservation of mammalian germ cells and embryos has been widely used in assisted reproduction. It also plays an important role in preservation of species diversity. DNA methylation abnormality induced by freezing in germ cells or embryos has an significant impact on the occurrence of disease and the physiological function of the offspring. In this paper, we summarize the effects of cryopreservation of mammalian germ cells and embryos on their pattern of DNA methylation which in turn make the developmental deficiency in the progeny from these gametes or embryos. Finally, it will provide the reference on the gamete cryopreservation and the development of assisted reproductive technology.

In this study,we used PCR-SCCP connect with ultrasonic vivo assay techniques, analysis of correlation between Leptin gene exon Ⅱ E2JW polymorphism and part meat quality traits of backfat thickness, eye muscle area, fat content, plot eye height in Xinjiang Brown cattle at ante-mortem, post-mortem. The results showed that Leptin gene exon Ⅱ E2JW site PIC was moderate polymorphisms (0.25 < PIC < 0.5), there were three genotypes of AA, AB and BB, AB genotype were the predominant genotype, B was the predominant allele. In addition, the forecast value of AB and AA gennotypes individual muscle fat content were inconsistent with actual value.Leptin gene exon Ⅱ E2JW was identified the theroy of a candidate gene loci of beef quality traits in Xinjiang Brown cattle.Genetic polymorphism and ultrasound measurements could not only fastly and accurately measure portion meat traits, but also as a means of marker-assisted selection.

The study was aimed to reveal the rule of growth and development and meat performance in Xiang White goose. 100 healthy Xiang White geese (50 male and 50 female) were selected and weighed during 0 to 12 weeks of age for study the growth and development by using Logistic model. 8 geese in each gender were slaughtered at age of 12-week and the body size index and meat yield performance were measured. The results showed as follows:① Xiang White goose grew rapidly in 1 to 6 weeks, and the increase of body weight was relatively stable during 7 to 12 weeks. After 4 weeks of age, the body weight of male goose were significantly (P<0.05) or extremely significantly (P<0.01) higher than the female. The fitting rates of Logistic model were over 0.988 in both male and female goose. The inflection of growth curves of female goose was at age of 5 weeks, and the male was 5.2 weeks. The correlation coefficient between the body weights at age of 4 weeks and 12 weeks were 0.512,0.561 in male and female goose, respectively (P<0.01).②The body length, keel length, tibia length and half-length diving of male goose at age of 12 weeks were significantly higher than the female (P<0.05).③The live body weight of male goose at age of 12 weeks was significantly greater than the female (P<0.05). ④The slaughter rate, eviscerated percentage, breast muscle rate, leg muscle rate of Xiang White goose were 88.25%, 67.93%, 13.03% and 13.76%, respectively. In conclusion, Xiang White goose grew fast, and had a good performance of meat production. The growth and development, body size and slaughter performance of male goose were better than the female.

In order to determine the drug resistance phenotype and prevalence of drug resistance genes in Escherichia coli (E.coli) isolated from sheep in Inner Mongolia, the minimum inhibitory concentrations (MICs) of the isolates to antibiotics commonly used in veterinary clinical were detected by micro-dilution method in vitro. The results showed that the highest resistance rates of the isolates to amoxicillin,cephalothin,sulfamethoxazole and polymyxin were up to 100.0%,respectively.To amoxicillin-clavulanic, tetracycline and ciprofloxacin were 50% to 80%. These isolates were sensitive to cefotaxime, meropenem trihydrate, neomycin, and their resistance rates were all less than 10%. Among the 108 strains of E.coli from sheep, 94.4% of them were resistant to more than 7 antimicrobial agents,15.6% of them were resistant to 13 antimicrobial agents, only one strain was sensitive to all antimicrobial agents. Six kinds of resistance genes among the 108 E.coli isolates were detected by PCR method.The results showed that detection rates of 4 kinds of drug resistance genes including bla_TEM, proP-2, sul-Ⅰ and ampG were all over 50%,the detection rate of resistance gene aph (3')-Ⅰ was up to 40%, only resistance gene aac(3)-Ⅱ detection rate was 5.5%. Thus, the sheep E.coli isolates in Inner Mongolia produced various degrees of resistance to 13 kinds of antibiotics, and their multi-drug resistances were very serious. They carried aph(3')-Ⅰ, sul-Ⅰ, ampG, bla_TEM, proP-2 and aac(3)-Ⅱ resistance genes.

In order to screen the host protein that interacted with σA gene of avian reovirus, the bait vector pGBKT7-σA of avian reovirus σA gene in yeast two-hybrid system was constructed. Total RNA of avian reovirus S1133 standard strain was purified by using Trizol reagents. Avian reovirus σA gene was amplified by RT-PCR and was cloned into yeast two-hybrid bait vector pGBKT7. The recombinant bait vector was named as pGBKT7-σA. The recombinant bait vector was transformed into Saccharomyces cerevisiae Y2HGold. And then the recombinant bait vector of different concentrations was applied to the nutrient deficient medium. The results showed that the yeast two-hybrid bait vector had been constructed successfully, and the recombinant bait vector pGBKT7-σA on yeast cells had no toxicity and self-activation. This study laid a foundation for further screening host protein interacted with σA protein by yeast two-hybrid technique.

To study the major structural protein glycoprotein (G) of infectious hematopoietic necrosis virus (IHNV), glycoprotein gene (1 380 bp) was amplified by RT-PCR from IHNV. In order to construct a recombinant plasmid pFB-LIC-Bse-G, G gene was cloned into the baculovirus vector pFB-LIC-Bse. Then, the constructed plasmid pFB-LIC-Bse-G was transformed into E.coli DH10Bac. The recombinant bacmids rBacmid-G was got, and then it was transfected to insect Sf9 cells,and the recombinant baculovirus that contained G gene was obtained. Western blotting analysis and indirect immunofluorescence assay (IFA) showed that the recombinant G protein could be recognized by histidine monoclonal antibody (Anti-His) and anti-IHNV antibody of mouse. The results indicated IHNV G protein had been expressed correctly in Sf9 cells. The study laid a foundation for further studying the G protein structure, function and immunological characteristics.

16 strains of Streptococcus were isolated and identified from 370 pig tissues of clinically morbidity farms and slaughter houses in Guangxi border area.Morphological observation, PCR identification and pathogenicity test were carried out. The results showed that the morphology,dyeing,biochemical characteristics were coincidence with the characteristics of Streptococcus suis,confirmed that 16 strains were Streptococcus suis,and named as GX1 to GX16. Further by divided type identification (35 serotypes) by GDH,1 strains was type 1,1 strains was type 2,1 strains was type 5,1 strain was type 29,5 strains were type 8 and 7 strains could not differentiate types. The pathogenicity of 16 strains Streptococcus suis were detected by Kunming mice and BALB/c mice. The results showed that the letal rate of GX10 for Kunming mice and BALB/c mice was 100%;GX11 for BALB/c mice was 80%;Other strains were apathogenicity to the Kunming mice and BALB/c mice. The LD₅₀ of GX10 and GX11 was 4.5×10⁵ and 3.6×10⁹ CFU for BALB/c mice,respectively. The results showed that pathogenic Streptococcus suis type 8 occured in Guangxi border area. The sensitivity of BALB/c mice to Streptococcus suis was higher than Kunming mice, BALB/c mice was more suitable for the pathogenic study of Streptococcus suis.

To obtain the monoclonal antibody (McAb) against VspX protein of Mycoplasma bovis (M. bovis),VspX gene was amplified, expressed and purified. Then, BALB/c mice were immunized subcutaneously three times with the purified recombinant VspX (rVspX) mixed with QuickAntibody-Mouse 5W adjuvant. Three days after the last injection, spleen cells were collected aseptically, and fused with SP2/0 myeloma cells in the presence of polyethylene glycol. By the clone selection, five stable hybridomas against VspX protein were obtained, separately named as 1A8, 3A3, 3C12, 3H9 and 4D11. Antibody titers in cell supernatant were from 1:1×10⁴ to 1:2×10⁵, while from 1:1×10⁵ to 1:8×10⁵ in ascites of mice by indirect ELISA. The subtypes were determined to be IgG1 and IgG2b class, and all light chains were κ chain. The affinity constant of McAb 3H9 and 4D11 were 6.3×10⁹ and 7.8×10⁹, respectively, and they belonged to high-affinity antibodies. Western blotting results showed that all of five McAbs could specifically react with M.bovis, however, McAb 4D11 could not react with Mycoplasma arginini PG2 and Mycoplasma mycoides subsp. PG3. Flow cytometry showed that McAb 4D11 reacted with surface VspX of M. bovis in a dose-dependent manner. Indirect immunofluorescence assay demonstrated that 4D11 McAb was able to detect rVspX protein binding to embryonic bovine lung cells. In the present study, McAbs against rVspX protein had been successfully prepared, which provided a basis for future researches about the function of VspX protein and the pathogenesis of M. bovis.

To establish a rapid method for differential detection of classical, highly pathogenic and TJM-F92 vaccine strains of North American genotype porcine reproductive and respiratory syndrome virus (PRRSV), a multiple RT-PCR assay was established. In this assay, two pairs of primers were designed according to the genomic sequences of classical, highly pathogenic and TJM-F92 vaccine strains of PRRSV. The assay could only detect PRRSV, but not detect CSFV, FMDV, PRV and PCV2. The detection limit of the method was as little as 1.13×10³ copies/μL of templates. The established assay was successfully used to detect 349 clinical samples and 119 samples were positive for PRRSV, of which 5 samples were positive for classical PRRSV (C-PRRSV), 107 samples for highly pathogenic PRRSV (HP-PRRSV) and 7 samples for TJM-F92 vaccine strain (V-PRRSV), while 7 samples were positive for HP-PRRSV and V-PRRSV. The results indicated that the established multiple RT-PCR assay could be used for differential detection and epidemiological investigation of PRRSV.

This experiment was to study the effect of synbiotic and antibacterial peptide on growth performance,and the immune regulation of broiler chickens after the E.coli O78 attack. Totally 100 broilers of health 1-day-old Ross 308 were randomly divided into four treatment groups:Synbiotic group (group S) was fed with basal diet+0.3% synbiotic;Antibacterial peptides group (group AP) was fed with basal diet+0.5% antibacterial peptide; Antibacterial peptide+synbiotic group (group SAP) was fed with basal diet+0.3% synbiotic+0.5% antibacterial peptide;Control group (group C) was fed with basal diet,and the growth performance were determined. At the 14th day,10 chickens from each group were randomly selected for muscle injection with E.coli O78 (the attacked control group was named as model group (group M)),and after 24 h,the morbidity,mortality and immune function of chickens in each group were determined. The results showed that compared with control group,the F/G and ADF of groups S and AP were significantly decreased (P<0.05).Before E.coli O78 attack,the bursa index of group SAP was significantly higher than control group (P<0.05),and other groups had no significant difference (P>0.05);While after the E.coli O78 attack,the thymus,spleen and bursa indexes of groups S and AP were significantly higher than control group (P<0.05),the spleen and bursa indexes of group SAP were significantly higher than control group (P<0.05),and the thymus index showed no significant difference (P>0.05).Compared with the group M,the IgA,sIgA and IL-10 of group S showed no significant difference (P>0.05),while the IL-6 levels decreased extremely significantly (P<0.01);The IgA and IL-6 of group AP were extremely significantly decreased (P<0.01),meanwhile the sIgA and IL-10 were significantly or extremely significantly higher (P<0.05;P<0.01); The changed trend of SAP group was the same as the group AP.In conclusion,the synbiotic and antibacterial peptide could improve the feed conversion rate of chickens and they were used alone or combination all could improved chicken immune function after the bacteria attack.

The objective of this study was to investigate the resistance of Salmonella and prevalence of resistance genes isolated from a pig farm in Xinjiang, and their coexistence with the major β-lactamases, 16S rRNA methylation enzyme genes and PMQR. The minimum inhibitory concentrations of Salmonella isolated from the pig farms were determined by agar dilution method, PCR was used to detect bla_TEM, bla_CMY-2, bla_CTX-M, bla_LAP-1, bla_KPC, bla_OXA and bla_SHV genes, 16S rRNA methylation enzyme genes armA and rmtB, and PMQR including qnrA, qnrB, qnrC, qnrD, qnrS, qepA, oqxA, oqxB and aac(6')-Ib genes. The positive strains were performed by using DNA sequencing to determine the purpose of the belt. The result showed that resistant rate of Salmonella isolates from swine were highest to ciprofloxacin and apramycin sulfate (77.9%, 102/131). Resistant mainly based on 8 kinds of drug resistance (29.0%, 38/131), 11 kinds of resistance genes were detection, the coexistence of different genotypes had 24 types. The main types were bla_TEM+bla_OXA+qnrS+aac(6')-Ib-cr+oqxA+oqxB (27.6%, 32/116), bla_TEM+bla_OXA+qnrS+oqxA+oqxB (24.1%, 28/116) and bla_TEM+qnrS (18.0%, 21/116). The Salmonella isolated from the farm had phenomenon seriously resistance, it coexisted with the main β-lactamase, 16S rRNA methylation enzyme genes and PMQR factors. The result suggested that it should strengthen monitoring to the β-lactamase enzymes, 16S rRNA methylation enzyme genes and PMQR factors.

The experiment was aimed to identify the bacterial distribution and drug resistance situation in deer farm environment form Xifeng county, Liaoning province. We collected manure, water, air and feed samples from deer farm for isolation and identification of bacteria, bacterial count, bacterial 16S rDNA PCR amplification, sequencing analysis, and the pathogenic bacteria and conditional pathogenic bacteria of drug sensitivity test. The results showed that there were 27 strains of bacteria isolated and purified, enumeration of male and female deer's pellet feed, fodder and water were 1.0×10⁵,8.7×10⁴,2.7×10⁷ CFU/g and 2.9×10⁴ CFU/mL, respectively. Description of water and fodder did not conform to the standard of the national health requirements; 16S rDNA gene sequences comparison in NCBI showed that these bacteria belonged to 15 kinds of different kinds, like Escherichia coli, Shigella sp., and so on. There were 13 kinds of pathogenic bacteria and conditional pathogenic bacteria. The 13 kinds were resistant to furazolidone, polymyxin B and cephalexin, and were sensitive to ceftriaxone, ciprofloxacin and amikacin. These results provided basis for efficient control of the disease in deer farm.

In order to obtain higher purity and high yield of LPS isolated from Escherichia coli, and evaluate its virulence,excrement of diarrhea piglet was collected as pathological samples, of which, some strains of Escherichia coli were isolated and identified. And furthermore, a strain of Escherichia coli was acquired by 16S rDNA identification. The LPS from this strain of Escherichia coli was isolated with the hot-phenol water method and purified by DNaseⅠ, RNaseA, proteinase K treatment and alcohol precipitation. The polysaccharide, protein and nucleic acid content of purified LPS were detected by phenol-sulfuric acid method, Bradford method and UV points spectrophotometric method. And its bioactivity and acute median lethal dose was determinated by tachypleus amebocyte lysate (TAL) method and acute toxicity experiment method, respectively. The results showed that a strain of highly pathogenic Escherichia coli was successfully isolated. The average yield of purified LPS was 3.74%, the proportion of the polysaccharide was 13.40%, the protein was 0.48%, and the nucleic acid was 0.06%. SDS-PAGE electrophoresis and silver stain showed that the bands were mainly concentrated in the range of 14 to 160 ku. The LAL bioactivity of the prepared LPS was 1.21×10⁷ EU/mg, LD₅₀ of the mouse abdominal cavity was 31.86 mg/kg. The results revealed that the diarrhea piglets in the farm was caused by pathogenic Escherichia coli,purified LPS from this strain of Escherichia coli had the advantages of higher purity, high yield and strong virulence, which would provide theoretical data for clinical prevention and treatment of diarrhea piglets.

A new feather-degrading bacterium was isolated from soil samples colected from pile-up feather places in this study.The transparent circle was produced after the strain was cultured on the milk medium for 24 h,the colony shape of the strain was irregular,rough surface,edge blur,and could produce green fluorescent pigment.It was gram negative,short rod,nospore and capsule and had a flagellum scanning by electron microscope. Biochemical test showed that the isolate could hydrolyze milk and gelatin,utilize citrate with keratinase activity and nitrate reduction reaction positive.Phylogenetic tree analysis showed that the isolate was most closely related to Pseudomonas aeruginosa strain (GenBank accession No.:BAMA01000316).Combined with the above results,the isolated strain was identified as Pseudomonas aeruginosa B1-2. When it was inoculated on basal medium containing 1% feather as sole carbon and nitrogen source,the highest level of keratinase activity was 60.3 U/mL after 48 h while the degradation rate of feather was 85.7%. This result indicated that the newly isolated Pseudomonas aeruginosa B1-2 could be useful in management of farm wastes.

In order to determine the safety of Bacillus amyloliquefaciens SSY1, we carried out the acute toxicity, chronic toxicity, bacterial translocation, product of metabolism and drug susceptibility test. Acute toxicity test in high dose groups (1.30×10¹⁰ CFU/mL), middle dose group(0.90×10¹⁰ CFU/mL), low dose group (0.65×10¹⁰ CFU/mL) and normal saline control group. After the acute toxicity test, the mice were sacrificed 14 days after the observation. Chronic toxicity test of each group was administered once a day for 30 d, chronic toxicity test in high dose groups (0.65×10¹⁰ CFU/mL), middle dose group (0.65×10⁹ CFU/mL), low dose group (0.65×10⁸ CFU/mL) and normal saline control group. The results showed that all mice were without exception phenomenon, the spirit, appetite, behavior and feces and so on, no pathological changes were found on the acute toxicity test. Chronic toxicity test of the mice were no abnormal clinical changes, and there were no pathological changes. Body weight and feed utilization rate of mice, hematology, blood biochemistry indexes and viscera coefficient difference were not significant (P>0.05);Bacillus amyloliquefaciens SSY1 strain did not occur translocation in mice, amino acid decarboxylase and indole test were negative, among the 24 kinds of commonly used drugs, the strains were sensitive to 23 kinds of drugs except the cillimycin. This experiment proved that Bacillus amyloliquefaciens SSY1 had good security.

56 cases of suspected tumor materials from a pet hospital in Beijing were collected from 2015 to 2016, the histopathological method was used for tumor diagnosis, then the varieties of onset, sites, age and gender of tumors were statically analyzed. The results showed that among the 56 cases, 44 cases were diagnosed as tumors. There were 21 cases of benign tumor, including rianalgland adenomas, sebaceous adenoma,mast cell tumor and so on. There were 23 cases of malignant tumors, including breast cancer, squamous cell cancer, adenocarcinoma of anus and so on. The results indicated that skin, perianal and breast had a higher incidence. The tumor onset rate became higher after 7 years old. The skin tumors and breast cancer were more common in female pet dog, however, the anal gland peripheral tumor onset rate of male pet dogs was higher. This study provided a reference for the epidemiology and diagnosis of canine tumors in the future.