This study was aimed to investigate the possible genes that involved in hormone regulation and regulate ovarian function, and selected G protein-coupled receptor 1 to in-depth study whether it was involved in hormone regulation.In this study, a pair of primers was designed based on the CDS sequence of GPR1 gene of Jinghai Yellow chicken which according to transcriptome sequencing.Real-time RT-PCR was used to detect the expression of GPR1 gene in the ovary of high and low-production Jinghai Yellow chicken. Bioinformatics analysis of GPR1 gene was performed by using combined biological softwares and online tools. The homology of GPR1 gene with other species, the physicochemical properties of protein, the transmembrane region, subcellular localization, hydrophilicity, potential phosphorylation site, conserved domain of protein sequence, secondary structure and tertiary structure of GPR1 protein were analyzed. The results showed that GPR1 gene of Jinghai Yellow chicken shared 100.0%, 69.0%, 69.1%, 69.3%, 68.9%, 69.6%, 69.2%, 68.8%, 68.9%, 67.5%, 69.0% and 50.3% identity with Gallus gallus, Homo sapiens, Bos taurus,Sus scrofa,Pan troglodytes,Leporidae,Equus caballus,Ovis aries, Pantholops hodgsonii, Rattus norvegicus,Mus musculus and Danio rerio, respectively.GPR1 protein structure showed that the molecular weight was 40.41 ku and pI was 6.91, including seven transmembrane segments which were consistent with the transmembrane analysis. Subcellular localization showed that GPR1 protein belonged to non-secretory protein. The predicted results showed that the protein contained 25 potential phosphorylation sites and 1 glycosylation sites. The secondary structure of GPR1 protein was mainly composed of random coil. The tertiary structure of domain area of GPR1 protein showed a transmembrane helix and single strand structure. The result of tissue expression showed that the expression of GPR1 gene in the low-production group was extremely significantly higher than the high-production group (P<0.01).The results of the study would be beneficial to the further analysis of the function of GPR1 gene, and provided a theoretical basis for the further study of the gene on the regulation mechanism of laying traits of Jinghai Yellow chicken.

To screen the reference genes of stable expression in different tissues of Hy-line layer (Gallus domesticus), RPS2, β-actin, GAPDH and HMBS genes were chosen as candidate gene, the software of geNorm, NormFinder and the Ct value were employed to analyze their stability in eight tissues (heart, liver, lung, kidney, duodenum, tibia, pancreas and uterus) of 120 days of age layers using Real-time quantitative PCR technology. The results showed that RPS2 gene was always dominant stability by the analysis of Ct value and geNorm software, the β-actin gene was followed; The NormFinder software showed that the HMBS gene was relative stability, and HMBS and RPS2 genes were the optimal combination; When analyzed the expression of constant reference gene in different tissues, it found that the most stable reference gene in different tissues was not the same. Therefore, it concluded that different tissues had different optimal gene, but they had the common potential, RPS2 and β-actin genes had the relative expression stability comparing the other gene.

The experiment was aimed to study the clone and prokaryotic expression of BPSS1512 gene in goat Burkholderia pseudomallei and analyzed its proteins by bioinformatics. The geneome of Burkholderia pseudomallei was used as the template,and the primers were designed by DNAMAN software referring to genomic DNA sequence of Burkholoderia pseudomallei K96243 strain in GenBank (NC_006351.1).The BPSS1512 gene was amplified by PCR and the recombinant plasmid was constructed. Then the expressed protein was analyzed by SDS-PAGE and Western blotting, and the amino acid sequence encoded by BPSS1512 gene was analyzed by softwares such as DNAMAN.The results showed that the BPSS1512 gene was successfully cloned with the length of 1 425 bp,and the recombinant plasmid pET-28a-BPSS1512 was constructed. The optimum conditions for induction was that the IPTG was 10 mmol/L and 8 h for induction.The molecular weight of the protein was 53 ku,it was expressed as the form of inclusion body.In the secondary structure of BPSS15122 protein,alpha-helix,extended strand,and random coil were 24.05%,14.77% and 61.18%, respectively,and the hydrophobic core was distributed between -2.0 and +2.4 which indicated that the BPSS1512 protein was strong hydrophobicity.

Gene editing techniques (GETs) can introduce mutations precisely into endogenous genome at a designed location. GETs are widely used in the research of bio-sciences, and promote a great development in life sciences. So far, most of the artificial gene mutations are generated by the following three GETs, including zinc finger nucleases (ZFN), transcription activator-like effector nucleases (TALEN) and clustered regularly interspaced short palindromic repeats/Cas endonuclease (CRISPR/Cas). GETs can recognize and modify the target sequence of endogenous gene with specificity, which are good ways to study the function of new genes. GETs have the advantage of simple operation, high efficiency and accuracy, so it is widely used in life science research, constraction of disease models, drug research and development and agricultural production. This review presents the three different nuclease-based GETs,including the mechanisms by which they produce genetic modifications, their recent advances and prospects as well as the differences between GETs and transgene techniques.

In order to detect Enterococcus faecalis endocarditis antigen (EfaA) of bear that was used to disclose the infected Northeast Black bear immediately, a pair of PCR primers was synthesized by EfaA gene of Enterococcus faecalis that was recorded in GenBank (accession number:U03756.1) and the target fragment was 689 bp. The PCR product was cloned by pMD19-T vector, then was transferred to Escherichia coli DH5α competent cells and the positive clones were filtered. Recombinant plasmid (pMD19-T-EfaA) was extracted. Identification of PCR and digestion, sequencing, the structure prediction of proteins were operated. The results showed that EfaA gene was cloned successfully, and the gene homology with ATCC 29212 was 100.0%. pMD19-T-EfaA was constructed successfully, structure of proteins was predicted. This study provided theoretical basis for diagnostic methods of Enterococcus faecalis and laid basis for prevention and control of Northeast Black bear disease.

This study was aimed to compare different culture methods for donkey skin fibroblasts to establish a highly efficient and rapid culture system. The dermal tissue of the healthy and 6-7 year old Xinjiang donkey was cultured for primary cells of skin fibroblast with tissue explants adherent method and double enzyme digestion method. Then,the growth characteristics,proliferation,cell cycle and mycoplasma contamination were detected and analyzed. The results showed that the fibroblast cells entered the logarithmic growth phase after cultured for 3 days by double enzyme digestion(0.25% trypsin digestion for 1 h and then 0.5% collagenase Ⅰ for 6 h), while it required 15 days with tissue explants adherent method. Flow cytometry analysis showed that nearly half of the cells were in G0/G1 stage,the other half in S+G2+M stage, while just 5.17% of the cells were apoptosis,which indicated that most of the cells were in the active stage of division and proliferation,and had strong vitality. Moreover, there was no mycoplasma contamination in the cultured cells. In summary,the culture system of donkey skin fibroblasts was rapidly and effectively established with the double enzyme digestion method in vitro.

This study was aimed to construct stable cell line expressing N protein coded by porcine epidemic diarrhea virus (PEDV) strain CH-HuN1301. N gene of this virus strain was amplified by RT-PCR, which was then cloned into prokaryotic expression vector pET28a and eukaryotic expression vector pEGFP-C1,respectively. Recombinant plasmids were sequenced and analyzed by software Mega 5.0. To prepare polyclonal antibody of N protein, the prokaryotic expression recombinant N protein was used as antigen to immunize rabbits. HEK293 transfected with recombinant eukaryotic expression plasmids pEGFP-PEDV-N was selected by G418 to produce a stable cell line which then was identified by immunoperoxidase monolayer assay (IPMA) and fluorescence observation. Results showed that, the N gene of PEDV strain CH-HuN1301 was 1 323 bp, molecular weight of recombinant prokaryotic expressing N protein was about 60 ku, prepared polyclonal antibody against N protein showed wonderful reactivity with PEDV propagated in Vero cells, recombinant eukaryotic expressing N protein was positively detected in the established stable cell line by IPMA and fluorescence observation. This stable cell line provided foundation for the research in PEDV diagnosis methods.

In order to explore the building conditions of acute lung injury, healthy Kunming mice were selected, by intraperitoneal injection of cascade dose of LPS (0, 2, 4, 6, 8, 10 mg/kg) to observe the mice respiratory frequency, mortality, lung W/D value and the changes of tissue structure in different conditions, and detect the expression changes of inflammation cytokine, IL-10,TNF-α, IL-1β and IFN-γ. The results showed that compared with control group, lung W/D value in 4 mg/kg LPS group significantly increased (P<0.05) and lung W/D values in 6, 8 and 10 mg/kg LPS groups extremely significantly increased (P<0.01). Pathological analysis showed that 6 mg/kg LPS group appeared lung congestion, alveolar luminal and pulmonary interstitial exudation, alveolar septal thickening, and a small amount of neutrophil infiltration. 8 and 10 mg/kg LPS groups appeared pulmonary congestion, alveolar and pulmonary interstitial exudation, alveolar septal thickening and neutrophil infiltration. Compared with control group, collagen fibers of the lung tissue in 6, 8, 10 mg/kg LPS group increased, and more appeared in the lung around the trachea, 10 mg/kg LPS group was the most obvious. The inflammatory factors in 2, 4, 6, 8 and 10 mg/kg LPS groups had extremely significantly increased compared with control group (P<0.01). Pathological biopsy and related indicators showed that the model of mice could be successfully constructed by intraperitoneal injection of 8 mg/kg LPS for 12 h.

The test was aimed to study the effect of insulin-like growth factor 1(IGF-1) on the proliferation of bovine mammary epithelial cells (BMECs),and provide a theoretical basis for the subsequent regulating mammary gland development using IGF-1. The optimal IGF-1 concentration for inhibiting BMECs apoptosis was obtained by measuring the apoptosis rate at 12, 24, 48 and 72 h after the addition of IGF-1 at four groups (0, 10, 50 and 100 μg/mL). Then divided into six groups:BMECs group, BMECs+IGF-1 group, BMECs+LY294002 group, BMECs+IGF-1+LY294002 group, BMECs+RAPA group and BMECs+IGF-1+RAPA group,and the apoptosis rates were detected by flow cytometry. The results showed that the heterologous IGF-1 had an inhibitory effect on the apoptosis of BMECs with an optimal concentration of 50 μg/mL. The apoptosis rates of BMECs+IGF-1+LY294002 and BMECs+IGF-1+RAPA groups were extremely significantly higher than BMECs+IGF-1 group (P<0.01). This study suggested that IGF-1 could activate the PI3K/Akt/mTOR signaling pathway and thereby inhibit the apoptosis of BMECs. Moreover, IGF-1 might also promote the "repair" mechanism for the inhibited PI3K/Akt/mTOR signaling pathway, and therefore make it reparticipate in BMECs life process.

In order to explore the influence of nucleoside diphosphate kinase B (NDPK B) on replication and proliferation of F48E9 strain of Newcastle disease virus (NDV), the eukaryotic expression plasmid pEGFP-NDPK B was transfected into Vero cells,then we got the cell subclones by G418 screening. We identified the expression of NDPK B protein by RT-PCR, Western blotting and inverted flurescence microscope. On the basis of Vero/NDPK B cell line,we explored the influence of NDPK B on replication and proliferation of F48E9 strain of NDV by HA-HI, TCID₅₀ and Real-time PCR.The results showed that we had successfully constructed the cell line named Vero/NDPK B, which could stably express NDPK B protein, and found that NDPK B could inhibit the replication and proliferation of F48E9 strain of NDV in Vero cells.It suggested that on the basis of NDPK B,we could design and develop new drugs or antiviral synergist to prevent or treat NDV.

In order to study the biological characteristics of somatic cells of Mashen pig, the ear marginal fibroblast cell lines of Mashen pig were established from 3 days old Mashen pig by direct explant method,and the biological characteristics of cell lines were detected. The results showed that the cell lines obtained from Mashen pig were in conformity with basic properties of typical fibroblasts, the shape of adherent cells were typical spindle, star and polygon, the time of grew into monolayer cells was 10 to 13 d, the growth curve was typical S shaped. The frequency of metaphase chromosome number (2n=38) was about 84% of the total number of cells, and it had met the establishment standard of fibroblast cell line. The transfection efficiency of green fluorescent protein(pEGFP-C1)transfected by lipofectamine-mediated method was 25%. The successful establishment of the fibroblast cell lines of Mashen pig had opened up a new method for the conservation of genetic resources.

The aim of the study was to research the effects of glutamic acid residue on growth and blood biochemical indexes of Qinchuan beef cattle. 12 bulls of 18-month-age,12 cows of 18-month-age and 12 bullocks of 8-month-age were selected and randomly allocated to 4 groups with 9 cattle per group (including 3 bulls,3 cows and 3 bullocks) in a single factor randomized block design.The control group and groupⅠ,Ⅱand Ⅲ were used 0,1.5%,3.0%,4.5% glutamic acid residue to replace part of soybean meal of diets. Feeding trial lasted for 100 d.Intake and body weight were measured at each stage during trial periord.At the end of the experiment, blood samples were collected to detect blood biochemical indexes.The results showed as follows:The average daily gain(ADG) of cattle in group Ⅰ,Ⅱ and Ⅲ were significantly higher than that of control group (P<0.05),and the value of group Ⅱ was significantly higher than that of group Ⅰ and Ⅲ (P<0.05). The dry matter intake (DMI) of cattle in group Ⅱand Ⅲ were significantly higher than that of control group (P<0.05). The feed/gain ratio (F/G) in group Ⅱwas significantly lower than that of control group (P<0.05).As for the blood biochemical indexes, the contents of alkaline phosphatase of control group, groupⅠand Ⅱwere significantly higher than that of group Ⅲ (P<0.05), and there were no significant differences for other biochemical indexes (P>0.05). The correlation between the average daily gain and the blood content of alkaline phosphatase was extremely significant (P<0.01). In conclusion,it could improve feed intake, daily gain (increased by 22.62%) and decrease the F/G (decreased by 10.85%) by using 3.0% glutamic acid residue replacing part of soybean meal in diets for Qinchuan beef cattle. Adding appropriate glutamic acid residue (1.5% to 3.0%)in diet could increase the level of blood alkaline phosphatase,which should be an important index to reflect the growth of Qinchuan beef cattle.

This study was conducted to study the effect of different ratios of concentrate to Pennisetum sinese Roxb in diet on in vitro rumen fermentation in Leizhou goats. In vitro fermentation method was used and diets with different concentrate to Pennisetum sinese Roxb were prepared which the ratio were 0:100 (group Ⅰ),10:90 (group Ⅱ),20:80 (group Ⅲ),30:70 (group Ⅳ) and 40:60 (group Ⅴ),respectively, and the samples were analyzed after fermented for at 4,8,12,24 and 48 h. The results showed that:①With the increase of the concentration to roughage ratios,the crude protein content of TMR diets increased and the fiber material and crude ash contents decreased gradually. With the increase of concentration,the gas production at 4,8,24,48 h were extremely significantly increased (P<0.01).②With the increase of fermentation time and the concentration to roughage ratios,the pH of rumen fluid were decreased,and pH of groups Ⅳ and Ⅴ were significantly lower than groups Ⅰand Ⅱ at the same time (P<0.05).And the dry matter degradation were increased,and that of groups Ⅳ and Ⅴ were extremely significantly higher than groups Ⅰand Ⅱ at the same time (P<0.01). The NH₃-N content at 48 h was higher than that at 4 h and there was no siginificant difference among groups at 4 and 24 h (P>0.05).③The correlation between pH and accumulated gas production, dry matter degradation rate were significantly or extremely significantly negative,that between accumulated gas production and dry matter degradation rate were significantly or extremely significantly positive (P<0.05;P<0.01).The correlation between pH and dry matter, crude ash, crude fiber, neutral detergent fiber and acid detergent fiber were significantly or extremely significantly positive,that with crude protein was significantly or extremely significantly negative (P<0.05;P<0.01).And the correlation between cumulated gas production,dry matter degradation rate and crude protein was significantly or extremely significantly positive,while that with the dry matter,crude ash,crude fiber, neutral detergent fiber and acid detergent fiber were significantly or extremely significantly negative (P<0.05;P<0.01). In summary, under the conditions of this test, the appropriate ratio of concentrate to Pennisetum sinese Roxb was 30:70 in which Leizhou goat could get a better fermentation effect and cost savings.

This study was conducted to examine the effects of cottonseed meal fermented by Lactobacillus acidophilus on antioxidant function and oxidation resistance of Yellow-feathered broilers. One hundred and eighty 21-day-old healthy male Yellow-feathered broilers were randomly divided into 3 groups with 6 replicates per group and 10 broilers per replicate.The broilers in the control group were fed the basal diet with 6% cottonseed meal supplementation,and those in groupⅠ were fed the experimental diet with 6% fermented cottonseed meal supplementation,and the others in group Ⅱ were fed the basal diet included 6% cottonseed meal and 7×10⁴ CFU/g Lactobacillus acidophilus.The results showed as follows:①At 42 days of age,the immune organ index of three groups were no significant difference (P>0.05).At 64 days of age,compared with control group, the spleen index, bursa of Fabricius index,serum IL-2 content and the expression of IL-10 mRNA in spleen of groupⅠwere increased by 47.69% (P<0.05), 20.59% (P<0.05), 5.77%(P<0.05) and 449.37%(P<0.01),respectively,and the expression of IFN-γ and IL-10 mRNA in spleen of groupⅡwere increased by 153.45% and 255.70% (P<0.05),respectively. The expression of IL-10 mRNA in spleen of group Ⅰ was increased by 54.45% compared with group Ⅱ (P>0.05).②At 42 days of age,the serum T-SOD,serum T-AOC and liver T-AOC of group Ⅰ were significantly increased by 8.26%,22.13% and 22.34% compared with control group (P<0.05),and the serum T-AOC of groupⅠwas significantly increased by 27.25% compared with groupⅡ (P<0.05).At 64 days of age,compared with control group,the serum T-SOD,serum GSH-Px and liver T-SOD of groupⅠincreased by 10.17%(P<0.01),20.45% (P<0.05) and 26.17% (P<0.01),respectively,and compared with group Ⅱ,the serum GSH-Px,liver T-SOD and liver GSH-Px of groupⅠincreased by 19.45% (P<0.05),18.41% (P<0.01) and 21.23% (P<0.05),respectively. In conclusion,dietary fermented cottonseed meal could enhance immune function and oxidation resistance of Yellow-feathered broilers, and Lactobacillus acidophilus had no significant influence on immune function and oxidation resistance of Yellow-feathered broilers.

The aim of this study was to select a strain of producing mannase Bacillus, and investigate its effects on growth performance of geese. Congo red plate changing color circle method and assay of enzyme activity method in liquid fermentation were used in screen and rescreen,respectively. Animal safety and gastrointestinal tolerability tests were alse carried out. By morphological observation,physiological and biochemical tests,analysis of complete sequence of 16S rDNA and the phylogenetic tree establishment,species were identified. Bacillus were prepared and added by 0.10%, 0.25%, 0.50%, 0.75% and 1.00% in the geese diets to study its effect on the daily gain and F/G in geese. The results showed that the enzyme activity of the isolated Bacillus subtilis Y-3-2 was 45.939 U/mL, and the strain could also endure pH 2.0 and grow normally in 0.1% cholic acid salt solution and artificial intestinal fluid. The results of feeding test showed that, compared to the control group, the average daily gain of geese increased by 8.78%(P<0.01) and F/G decreased by 6.69% (P<0.01) by adding 0.25% Y-3-2 preparation. In conclusion, adding mannase producing Bacillus could improve the feed utilization and growth of geese.

Threonine (Thr) is one of the four main limiting amino acids of poultry, and it plays a very important role in poultry's maintenance of normal growth, development and immune function. Although there have been very comprehensive studies on the mechanism of broilers and laying hens over the past ten years, there are still relatively fewer studies on ducks. This paper summarizes recent studies on Thr nutrition-physiological function in broilers, laying hens and ducks, including the influence of Thr on dietary amino acids balance, production performance, carcass quality, immune function and intestine health maintenance. At the same time, this paper also briefly analyzes the factors related to Thr requirement, including growth stage, crude protein level, energy concentration, other amino acids level in diet and raising environment. In brief, the review is respected to guide poultry production practices.

The purpose of this study was to investigate the property of antibacterial, production of enzymes, tolerance and in vitro digestibility of 11 Bacillus subtilis strains.9 Bacillus subtilis strains were screened out according to antibacterial property and production of enzymes. After tests of heat, pH tolerance, artificial gastrointestinal fluid and pig bile salt,6 Bacillus subtilis strains had good performance in simulation of animal digestive tract and strong heat tolerance in the process of production. Meanwhile, 5 strains BLCC1-1, BLCC1-2, BLCC1-3,BLCC1-4 and BLCC1-6 had good digestion effects on different combinations of feeds. Based on the above results,it suggested that BLCC1-1, BLCC1-2, BLCC1-4 and BLCC1-6 had excellent production of enzymes, tolerance and in vitro digestibility, which might be suitable for using as feed additives and further research.

This experiment was aimed to study the effects of protein hydrolysate on growth performance,blood biochemical indexes and nutrient apparent digestibility in growing-finishing pigs,and determine the optimum dosage of additions.300 Doroc pig×Landrace pig×Yorkshire pig crossbreeding commercial pigs were chosen and divided into five groups with four repeats per groups and 15 pigs per repeat.The pigs in group Ⅰ (control group) were fed with basal diet,others in groups Ⅱ to Ⅴ were fed basal diet with 1%,2%,3% and 4% T30 small peptide,respectively.The test lasted for 30 d. The results showed that:①The average daily gain (ADG) of group Ⅲ was significantly higher than groups Ⅰ and Ⅴ (P<0.05), the average daily feed intake (ADFI) of group Ⅲ was significantly higher than group Ⅱ, and had no significant difference with other groups (P>0.05). ②The total protein and globulin levels of group Ⅴ were the highest that significantly higher than group Ⅱ (P<0.05),and had no significant difference with other groups (P>0.05).The urea nitrogen of group Ⅱ was significantly lower than other groups (P<0.05).③The dry matter and crude protein apparent digestibility of group Ⅴ were the highest,and the Ca and P apparent digestibility were declined in T30 supplement groups. In conclusion,the optimum adding amount of protein hydrolysate of growing pig was 1.796%.It could improve the growth performance,decrease F/G,and increase the levels of total protein and globulin.

This study was aimed to study the effects of tomato pomace on growth performance,antioxidant capacity and economic benefit of finishing pigs.80 Doroc pig×Landrace pig×Yorkshire pig crossbreeding finishing pigs with (95.20±3.95) kg body weight were chosen and divided into four groups with four repeats per groups and 5 pigs per repeat. The pigs in control group were fed with basal diet,others in groups Ⅰ,Ⅱ and Ⅲ were fed basal diet with 100,200 and 300 g/d tomato pomace supplementation,respectively. The test lasted for 37 d.The results showed that compared with control group,the ADG and ADFI of group Ⅰ were increased by 8.93% and 18.81%(P<0.01),respectively,while the F/G of group Ⅰ was decreased by 11.03% than control group (P<0.05).The nutrient apparent digestibility of dry matter and crude ash in groupⅡ were increased by 5.66% (P<0.05) and 14.81% (P<0.01),respectively. The crude fiber apparent digestibility of groups Ⅱand Ⅲ were significantly reduced by 9.25% and 11.88% (P<0.01),while the digestibility of crude protein and crude fat had no significant difference among the four groups (P>0.05).The SOD activity of groups Ⅱ and Ⅲ were significantly increased by 40.99% and 36.54% (P<0.01),respectively,and the MDA concent in groups Ⅰ,Ⅱ and Ⅲ were significantly decreased (P<0.05), while the serum T-AOC and GSH activity had no significant difference among the four groups (P>0.05).The weight gain income of group Ⅰ was 586.26 yuan/head,increased by 48.06,52.20 and 52.92 yuan than control group,groups Ⅰ and Ⅱ,respectively. The profit of groupⅠ was the highest (466.90 yuan).In conclusion,tomato pomace supplementation could improve growth performance,nutrient apparent digestibility,antioxidant capacity and the economic benefit,but the content should be controlled.

This experiment was conducted to investigate the effects of antimicrobial peptide on growth performance and immune function of 817 broiler hybrids. Nine hundred one-day-old 817 broiler hybrids were chosen and randomly divided into 5 groups with 6 replicates per group and 30 broilers per replicate.The broilers in control group were fed with basal diet, broilers in the antibiotic group were fed with basal diet supplemented 150 mg/kg aureomycin,and that in groups Ⅰ,Ⅱ and Ⅲ were fed with basal diet supplemented 100,200 and 300 mg/kg antimicrobial peptide,respectively. The experiment period was 42 days. The results showed that average daily gain of 817 broiler hybrids in groups Ⅱ and Ⅲ was significantly increased (P<0.05),meanwhile the F/G was significantly decreased at 1-21 days old (P<0.05).The immune organ indexes of thymus,bursa of Fabricius and spleen were significantly increased at 21 and 42 days old (P<0.05),and the antibody level against ND and T lymphocyte transformation rate of 42 days old 817 broiler hybrids were improved (P<0.05). It indicated that antimicrobial peptide could increased growth performance and immune function of 817 broiler hybrids as same with aureomycin,and appropriate dosage of antimicrobial peptide was 200 mg/kg in this experiment.

In order to explore the genetic diversity and the classification status of the Chinese domestic pigs at the molecular level, the complete mitochondrial DNA (mtDNA) genomic sequences of 22 Chinese indigenous pig breeds from 6 types were downloaded from GenBank for the analysis using the bioinformatics method. The polymorphism of nucleic acid sequences was analyzed,the molecular phylogenetic tree based on D-loop sequences, Cytb gene, complete coding region mtDNA sequences and the Median-Joining network of the mtDNA D-loop haplotypes were constructed for 22 pig breeds from 6 types. The results showed that a total of 144 mutation sites among 22 pig breeds were detected, 22 haplotypes were discovered, which indicated that Chinese indigenous pig breeds had abundant genetic diversity. According to the complete mtDNA genomic sequences analysis, which suggested that the main mutations was transition, the transition/transversion ratio was greater than 2.0 and all mutations were in line with the neutral mutation. Our findings clearly demonstrated that there was a near genetic distance among 6 types, meanwhile, the shared haplotypes were found. The phylogenetic tree showed that Chinese indigenous pigs from 6 types were diverged from two ancestors.The results indicated that mtDNA D-loop and Cytb gene might serve as molecular markers for phylogenesis, origin and evolution.

This study was aimed to elucidate the molecular mecbanisms of Smad4 gene on granulose cells proliferation and embryonic development in buffalo. The Smad4 gene was amplified by RT-PCR, analyzed by bioinformatics, studied with eukaryotic vector construction, and used liposome transfection skill to transfect into the buffalo granulose cells. The results showed that Smad4 gene was cloned, the coding region was 1 662 bp, and encoded 553 amino acids. BLAST analysis showed that the buffalo Smad4 gene shared 99% of similar nucleotide sequence with Bos taurus, and shared 98%, 96%, 96% and 95% of similar nucleotide sequence with Ovis aries, Sus scrofa, Equues caballus and Homo sapiens, respectively. Phylogenetic tree analysis showed that Smad4 gene was highly conserved in different species. The buffalo pEGFP-N1-Smad4 eukaryotic expression vector was successfully constructed, and transferred into the buffalo granulose cells, and the Smad4-EGFP fusion protein was detected in the cells. The results suggested that the success cloning and construction vector of buffalo Smad4 gene laid the foundation for the research of Smad4 gene on embryo development.

This study was aimed to analyze the association of insulin-like growth factor 1 (IGF1) gene polymorphisms with growth traits in rabbit. The technology of DNA pooling, PCR-SSCP and direct sequencing were used to detect the SNP of IGF1 gene. The results showed that T365C was detected in IGF1 gene exon 3 of rabbit. The SNP (C/T) was devised into three genotypes:TT, TC and CC. The relationships between genotypes and growth traits revealed that the birth weight and the weight at 28 days of age in New Zealand rabbit of TT genotype of T365C locus were significantly higher than TC and CC genotypes (P<0.05), the birth weight, the weight at 90 days of age and average daily gain form birth to 90 days of age in Belgian rabbit of TT genotype of T365C locus were significantly higher than TC and CC genotypes (P<0.05), there were no significant difference of the growth traits in other rabbit breeds (P>0.05).The consequence indicated that IGF1 gene was primarily deduced to be a potential major gene or linked to major gene effecting the growth traits of rabbit, and this SNP (T365C) might be a candidate molecular genetic markers to improve the growth traits of rabbit.

Multi-vertebra trait means increase of the vertebral number in animal, which is positive for livestock. Increased thoracic-lumbar vertebral number will result in improvement of carcass traits, especially carcass length, as well as economic value. Meanwhile, relatively high heritability of multi-vertebra trait is evaluated, providing genetic basis for selecting and breeding of multi-vertebra livestock. As such, individuals with multi-vertebra trait will increase production of domestic animals ultimately. Thus, study on multi-vertebra trait provides theoretical basis for breeding of meat livestock with multi-vertebra. Hitherto, multi-vertebra trait has been found in swine, sheep and bovine. Much information has been revealed in studies on multi-vertebra trait in swine; While that is limited in sheep and bovine. This review summarized variations of vertebral number, effects of multi-vertebra trait on the production traits, and genes or mutations responsible for multi-vertebra trait of livestock animal. Further, the research gap and future direction were analyzed as well.

To further optimize and promote the reproductive performance of the domestic level on sheep breeding,this paper focus on bone morphogenetic protein 15 (BMP15) gene, which is a major gene of reproductive traits in recent research,and combing its research progress, direction and pathway. In this paper,its signal transduction, mechanism, performance of sheep breeding and effect of production research in domestic practice were discussed. Totally,the domestic research of BMP15 gene is not deep enough in the mechanism of action, or in the marker assisted selection of applications filed that needs more research scholars to participate in this work. Meanwhile to provide reference of the research of BMP15 gene and marker assisted selection of domestic sheep.

A Bacillus strains,MY-6 strain which secreted high amount of protease was isolated from healthy pig's intestines,and it was identified as Bacillus amyloliquefaciens by the methods of morphological observation, biochemical tests,16S rDNA sequence amplification and comparison, biolog identification. In order to get an initial understanding of the enzymatic properties of its protease, the dynamic relationship between protease activity and bacterial life cycle, effects of pH,temperature,metal ions and inhibitors on protease activity,and the protease hydrolysis of different substrates were investigated. The results showed that,the protease synthesis of MY-6 strain was synchronized with its spore formation which began in the early stage of the exponential growth phase. The optimum conditions for the protease was 40℃ and pH 7.0,Ca²⁺ and Mn²⁺ had a intensive effects on the protease activity, PMSF could completely inhibit the activity of protease, indicating that the protease was a serine protease.The content of TCA soluble peptides for protein hydrolyzate fish protein, soybean protein, corn protein and casein was 0.83,0.67,0.58 and 0.38 μmol/g,respectively. In conclusion,Bacillus amyloliquefaciens MY-6 strain could be used as an initial selection of probiotic Bacillus to improve protein digestion.

To develop an effective vaccine against porcine circovirus type 2 (PCV2), ORF2 gene was cloned into a Lactobacillus plantarum (L.plantarum) pSCPSP under the control of a SCP promoter to construct recombinant plasmid pSCPSP-Cap that was transformed into LP1 by electroporation. Extracellular expression of the Cap protein was confirmed by SDS-PAGE and Western blotting analysis. Afterwards, 8-week-old SPF BALB/c mice were randomly divided into control group, carrier group, lavage group, group of drinking water and inactived vaccine group. After 14,28,42 and 56 d, mice serums were collected to evaluate the level of specific antibody by SDS-PAGE and Western blotting. Eventually, SDS-PAGE and Western blotting results indicated that the Cap protein of PCV2 could be efficiently expressed in L. plantarum, expression of protein molecular weight was 28 ku and with good immunogenicity. In addition, higher levels of specific antibody in the serum of mice was observed by oral administration with L. plantarum expressing the PCV2 Cap antigen (P<0.05). In conclusion, the ORF2 gene of PCV2 could be successfully expressed in recombinant L. plantarum and expressed protein with good biological activity, which would lay the foundation for the development of PCV2 oral vaccination.

We tried to identify the bacteria and explore the mechanism of the bacteria's pathogenicity via housekeeping gene gyrB and in vitro organ culture (IVOC) of ileum and intramuscular injection. Microscope, electron microscope and scanning electron microscope were also used to observe and the structure, pathogen of M12 and changes of infected tissues. The results showed that M12 was short gram-negative bacteria, and both ends of it were obtuse and the size without flagellum was (0.6 to 1.6) μm×(0.6 to 0.7)μm while length of flagellum was about 2 to 3 times of the length of bacteria.It had 100.0% similarity with Aeromonas caviae from GenBank. IVOC test observed M12' adhesion in intestinal epithelial cell, resulting in formulation of biofilm structure and damage in intestinal epithelial cell. Histopathologic examination showed that M12 could cause damage in intestine, liver, lung, kidney, muscle tissue of rabbit.The assay would offer references for researching the pathogenic mechanism of Aeromonas caviae.

This study was aimed to explore the pathogenesis of calf diarrhea in a large scale dairy farm in Shanxi province and the correspond comprehensive prevention and control measures. By investigating the incidence of calf diarrhea in the field, rotavirus, coronavirus,E. coli F5 (K99),Cryptosporidium parvum, coccidia and Salmonella were detected through experimental diagnosis, and the suspicious pathogenic bacteria was isolated and cultivated,which was identified with morphology and serology, simultaneously sensitive antibiotics were screened according to the antibiotic susceptibility tests, which contained 18 kinds of common antibiotics. The results showed that the clinical diagnosis and laboratory tests basically ruled out the possibility of E. coli F5 (K99), rotavirus, coronavirus, Cryptosporidium parvumand coccidia infection, and the isolated suspected bacteria had Salmonella colony characteristics and bacterial morphology, which showed positive reaction to Salmonella O polyvalent A-F serum, so we had identified that it was Salmonella diarrhea.The antibiotic susceptibility tests results showed that Salmonella was highly sensitive to ceftriaxone, gentamicin, cephradine and amikacin, which could be recommended for clinical treatment. Above all, this study proposed the specific prevention and treatment of the disease, and better control effect was achieved. This study initially provided a reference for the diagnosis and treatment of Salmonella of calf diarrhea, with great clinical significance in the early diagnosis and treatment of pathogens of calf diarrhea.

This study was aimed to explore the effect,dose of Lactobacillus reuteri on inhibition of pathogenic Escherichia coli infection on Kunming mice as well as its potential mechanism. 96 Kunming mice were randomly allocated into four groups with 4 replicates per group and 6 mice per replicate,which were control group,low dose group,middle dose group and high dose group. All of the mice feed commercial basal feedstuff,meanwhile the mice in control group drank tap-water,three treatments were supplemented with 1.0×10⁶,1.0×10⁷ and 1.0×10⁸ CFU/mL Lactobacillus reuteri in water. The feeding period was 28 days. All the mice were weighted, and the ADFI,ADG and F/G were calculated at the end of experiment.At the day of 28,pathogenic Escherichia coli (4.98×10⁹ CFU/mL) were irrigated to study the mortality. The indexes of intestinotoxin and antioxidation in serum and cecal microbacteria were inspected.The results indicated that supplemented with middle dose of Lactobacillus reuteri could significantly improve the final body weight,ADG compared with the control (P<0.05). The mortality, serum intestinotoxin level and MDA concentration,Escherichia coli and Staphylococcus were significantly lower than control group (P<0.05),while the T-SOD,T-AOC,GSH-Px,Lacobacillus and Bifidobacterium were significantly improved (P<0.05).In conclusion,with Lactobacillus reuteri supplementation,the growth performance was improved and antioxidant capacity and cecal microbacteria were enhanced after pathogenic Escherichia coli infected, mouse mortality caused by the pathogenic Escherichia coli was effectively reduced,and the effect of 1.0×10⁷ CFU/mL Lactobacillus reuteri was the best.

This study was aimed to prepare excellent fluorescence immunochromatographic strip for clenbuterol (CLB),ractopamine (RAC) and salbutamol (SAL),which could achieve the purpose of rapid and quantitative detection and laid a foundation for its industrialization. The properties of this strip including detection rage, detection limit, accuracy, degree of precision, false positive, false negative and coincidence with instrument method were investigated. The standard curve range of CLB, RAC and SAL test strips were 0 to 3.2, 0 to 6.4 and 0 to 8.0 μg/L, respectively.The detection limit of CLB, RAC and SAL were up to 0.2,0.4 and 0.5 μg/L, respectively. The cross reaction between CLB,RAC,SAL and other eight similar drugs were negative which demonstrated the good specificity. The added recoveries of each batch were between 60% and 120%,which met the requirements.The coefficients of variation(CV)under different concentrations of CLB,RAC and SAL were less than 15%, which met the precision standard.Besides,the coincidence with instrument method was 100%. The strips was suitable for rapid detection of CLB, RAC and SAL residues in pig urine with the advantages of high sensitivity, good stability and strong specificity.

In this study,tdh-vvhA-ctB was constructed using the flexible Linker sequence (Gly₄Ser) in order to construct three kinds of food-poisoning Vibrio poly-recombinant toxin gene and recombinant expression vector. The recombinant expression plasmid pET-22b(+)-TVC was constructed and expressed in prokaryotic expression vector. The animals were immunized using the expressed protein after purification to get serum antibody. The specificity and sensitivity of the antibody were verified by agar diffusion test and enzyme-linked immunosorbent assay (ELISA).The results showed that the recombinant expressing plasmid pET-22b(+)-TVC was constructed and expressed successfully in prokaryotic expression vector, the expression level was 11.38%. The expressed protein was mainly inclusion body and a small amount of soluble protein. The gene length was 2 196 bp, encoding 731 amino acids with molecular weight of 81.7 ku. The results were 99.6% homologous to the designed sequence. Agar diffusion reaction and ELISA tests indicated the poly-recombinant toxin TVC could product different immune intersect reaction with other food-poisoning Vibrios but not react with some no-objective bacteria. Expression plasmid of poly-recombinant toxin gene was constructed and serum antibody was prepared successfully. It might be used to check the objective toxin based on the poly-recombinant toxin gene and established the board-spectrum, quick, special detecting way to lay the theoretical and technical basis.

In order to explore the optimal immune procedure of foot-and-mouth disease (FMD), classical swine fever (CSF) and highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) vaccines in backyard pigs in rural areas, field immune experiment was carried out in two towns. Town A used "332" immune procedure, namely, FMD vaccine and CSF vaccine were simultaneously injected at different sides of the neck, and HP-PRRS vaccine was injected 14 days later. Town B used "321" immune procedure, namely, HP-PRRS vaccine and CSF vaccine were mixed together and injected in one side of the neck, and FMD vaccine was injected in the other side of the neck. FMD, CSF and HP-PRRS antibodies were detected by ELISA, and FMDV, CSFV and HP-PRRSV were detected by RT-PCR. The assessment indexes, including the immune density, immune side-reaction rate, death rate of pigs, positive rate of antibody of backyard pigs, positive rate of antibody of slaughter pigs, positive rate of pathogen of backyard pigs, positive rate of pathogen of slaughter pigs and so on, were collected and analyzed. The results showed that, compared with town A used "332" immune procedure, the HP-PRRS immune density, the positive rate of CSF and HP-PRRS antibody of backyard pigs, the positive rate of HP-PRRS antibody of slaughter pigs were significantly increased (P<0.05), while the death rate and the positive rate of HP-PRRSV of slaughter pigs were significantly decreased (P<0.05) in town B used "321" immune procedure. The other major assessment indexes had no significant difference (P>0.05). The results indicated that the immune efficacy of "321" immune procedure was better than that of "332", and it was worthy to be applied in clinical practice.

Staphylococcus aureus (S.aureus) is a gram-positive bacteria which could cause many kinds of infections, such as arthritis, endocarditis cellulitis, septicemia, pyemia and other systemic infections. In recent years, due to the unreasonable use of antibiotics, resistant Staphylococcus aureus increased rapidly.Particularly, the oxacillin-susceptible methicillin-resistant Staphylococcus aureus (OS-MRSA),it is a great challenge for clinical treatment. Understand the epidemiological characteristics and the mechanism of drug resistance of OS-MRSA can help clinicians to choose therapeutic drugs much better. This review summarized the research progress of epidemiology and resistant mechanism of OS-MRSA.

The aim of this study was to establish the quality standard of Ruxiankang (RXK) injection. According to the relevant requirements of the Chinese Pharmacopoeia (2015 edition), TLC identification,related substances examination and heavy metal examination method of RXK injection were established and the limits were determined. HPLC method was used to establish the content determination method of liquiritin, ammonium glycyrrhizinate, hesperidin and caffeic acid, and determined the limit. The specificity of Glycyrrhizae Radix et Rhizoma, Citri Reticulatae Pericarpium and Taraxaci Herba was strong. Liquiritin in 0.0459 to 0.7340 μg, ammonium glycyrrhizate in 0.0661 to 1.0580 μg, hesperidin in 0.0592 to 0.9470 μg, caffeic acid in the 0.0384 to 0.6140 μg concentration showed a good linear relationship with peak area. The correlation coefficients were 0.997,0.9990,0.9999 and 1.000,respectively. The average recoveries were 101.25%, 99.87%, 98.39% and 103.49%,respectively. The RSD values were 4.82%, 3.63%, 2.94% and 0.85%,respectively. The contents of liquiritin, ammonium glycyrrhizinate, hesperidin and caffeic acid of RXK injection were not less than 88.90, 138.48, 136.50 and 21.54 μg/mL,respectively. According to the maximum daily dose of RXK injection, Pb,Cd, As, Hg and Cu should not exceed 23,3,6,2 and 150 μg, respectively. The quality standard established in this study could provide reference for controlling the quality of RXK injection.

For preparation of bioactive single chain fragment variable (ScFv) which was targeted against envelope glycoprotein D (gD) of infectious bovine rhinotracheitis virus (IBRV),VH and VL genes were amplified from the cDNA of lymphocyte hybridoma, then VH and VL genes were integrated with a Linker by SOE-PCR,and the ScFv gene was cloned into the baculovirus vector pFB-LIC-Bse, which was transformed into Escherichia coli DH10Bac competent cells to construct a recombinant bacmid rBacmid-VH-VL. Finally,ScFv was expressed by transfected rBacmid-VH-VL into insect Sf9 cells, its molecular weight was about 30 ku. The result of Western blotting suggested that ScFv generated in Sf9 cells was specifically binds to His antibody,the protein also showed high affinity to gD of IBRV. The result of indirect immunofluorescence assay showed that ScFv could recognize IBRV which infected bovine kidney cells (MDBK). The study indicated that the recombinant ScFv was expressed in Sf9 cells, and could bind to gD of IBRV specifically. The result could provide the basis for the diagnosis and treatment of infection of IBRV.

This study was aimed to develop a method for simultaneous determination of five kinds of penicillins in eggs by high performance liquid chromatography-ultraviolet detection (HPLC-UVD). The eggs samples were extracted by acetonitrile,degreased by n-hexane,concentrated by rotary evaporation,derivatization,and then was separated by HPLC. The results showed that the calibration curves of five kinds of penicillins were linear correlated in the concentration ranges of 10 to 1 000 μg/kg with the correlation coefficients >0.999,which the limits of detection and quantification were 1.5 μg/kg (S/N=3) and 5.0 μg/kg (S/N=10),respectively. The average recoveries were 62.04% to 93.13%,the relative standard deviations were 3.91% to 13.69%.And the intra-day and inter-day RSDs were 3.98% to 14.71% and 4.26% to 12.71%,respectively. The developed method was suitable for detecting five kinds of penicillins residues in eggs with high repeatability and sensitivity.

The harm for livestock and poultry production caused by Mycoplasma infection has got widespread attention of researchers in the world. The practical ways to control the disease in order to reduce economic loss through Mycoplasma infection is the extensive use of antibiotics. Many groups of antibiotics including tetracyclines, macrolides and quinolones have been shown to be effective to Mycoplasma. Clinical practice shows many Mycoplasma isolated from livestock and poultry are characterized not only by single drug resistance but also by multiple antibiotic resistance due to the legacy of past decades of antimicrobial misuse, which threat the livestock and poultry production health. This paper reviews the mechanism of anti-Mycoplasma medicines through the alterations in the targets, the formation of efflux pump system, and the generation of inactivated enzymes of antibiotics, which provide theoretical basis and strategies for establishing resistance testing system, rational drug use and development of new drugs.

An endophyte strain producing β-mannanase was isolated and identified from sprouting konjac seeds,then enzymatic characteristics of β-mannanase was studied. The highest β-mannanase producing strain was obtained by transparent circle preliminary screening and shake flask fermentation secondary screening,and the enzyme activity was evaluated according to dinitrosalicylic acid (DNS) method,and then the strain was identified based on 16S rDNA sequence. The results showed that it was most closely related to Enterobacter ludwigii (99% sequence similarity). The optimal temperature of the β-mannanase was 60℃,and it had a good stability under temperature from 30 to 50℃. The optimal pH of the β-mannanase was 6.0,and it was stable over a range of pH 4.0 to 9.0. The enzyme was slightly activated by Zn²⁺(117.84%),EDTA(115.80%),Cu²⁺(113.76%). On the contrary,Mn²⁺(22.02%) could strongly inhibit the enzyme activity. The Km of the β-mannanase towards konjac flour was 26.65 mg/mL. Furthermore,after 72 h shaking-flask fermentation,the β-mannanase activity achieved 9.48 U/mL,and it had excellent enzymatic characteristics. These superior properties made the β-mannanase a broad application prospects in feed industry.

In order to explore the making procedure of vascular corrosion casts of ovarian artery in sheep and study the distribution of the blood vessels, this study was carried out and it would provide the foundation of blood vessels anatomy of reproductive system including blood supply mechanism of female ovaries in mammals. Firstly, samples of the uterus and appendages in sheep were collected, and then ovarian arteries and their branches were infused with 8% ABS perfusion agents. After the agent became solid, the samples' soft tissues were corroded away with 30% hydrochloric acid, washed them down with running water and the moderate trim process was done for obtaining the complete vascular corrosion casts of ovarian artery in sheep.The finished casts showed that the ovarian artery in sheep arises three branches including uterine, ovarian and tubal. The ovarian branch showed complicated arrangement such as the spiral artery of ovarian hilus, light spiral piece of ovarian hilus and tight spiral piece of ovarian branch. The results confirmed that the ovarian branch was characterized by the presence of a tight spiral configuration in sheep, the ovarian arterioles were fasten on ovarian hilus, and there were some dense capillaries which extend to other regions of the ovary followed them. Through this procedure, the vascular distribution of corrosion casts was clear,three-dimensional and with smooth surface. It could provide a reference or model for the study of ovarian blood vessels in mammals.

Fluoroquinolones (FQs) as one kind of antibiotics, are widely used for prophylaxis and treatment of intestinal diseases, respiratory illnesses and so on. However, the significant progressive increase of their use caused inevitable residues in food that represent a potential health hazard for consumers and drug resistance bacteria. Therefore, it is important to find a fast and reliable analytical method for sensitive determination of fluoroquinolones. Detection objects include fluoroquinolones in food, blood and urine. This review summarized the recent progress of the latest methods for detecting fluoroquinolones over the last three years, which including electrochemical method, high performance liquid chromatography, high performance liquid chromatography-mass spectrometry, enzyme linked immunosorbent assay, fluorescence polarization immunoassay, capillary electrophoretic immunoassay-laser induced fluorescence, upconversion fluorescence resonance energy transfer immunoassay, surface plasmon resonance sensor, thin layer chromatography and so on. The advantages and disadvantages of the above methods are also analysed in this review. Furthermore, the development trend of the methods for the determination of fluoroquinolones is discussed.

To establish the reversed phase high-performance liquid chromatography (RP-HPLC) method for determination of plasma concentration of acetaminophen in Bactrian camel, the chromatography was carried on a C18 column (150 mm×4.6 mm,5 μm).The mobile phase was composed of methanol-water (20:80),the flow rate was 0.5 mL/min,and the detection wavelength was set at 248 nm for acetaminophen.The retention time of acetaminophen was 6.5 min, and was not disturbed by other miscellaneous peaks.The calibration curve was linearity in the range of 0.08 to 10.24 μg/mL (R²=0.9997),and the quantitation limit of acetaminophen was 0.10 μg/mL.Both the inter-day and intra-day RSD were under 7.0%, the absolute recovery of the method was 80.39% to 93.56%,the relative recovery of the method was 94.66% to 104.73%.Therefore, the established method was simple, reproducible and accurate,which could be used for determination of plasma acetaminophen concentration and in vivo activities of CYP1A enzyme in Bactrian camel.

The purpose of this study was to evaluate the safety of the cefquinome sulfate liposomes by using the acute toxicity test, local irritation test and chronic toxicity test. The results showed that the LD₅₀ of cefquinome sulfate liposomes was 639.73 mg/kg after intramuscular injection of its suspension and 95% confidence interval was 573.06 to 714.17 mg/kg. There was no significant stimulation to muscle. Subchronic toxicity test showed that the weight, haematological indexes and biochemical criterion of rabbits in variety dose had no significant difference compared with normal group (P>0.05). There was no visible pathological changes on the liver,kidney histology. According to pathological observation, the structure of liver and kidney were clear in treated and middle dose groups,but there existed disorganized hepatic cell cord in the liver and hemorrhag between glomerulopathy,tubulointerstitial and hepatic sinusoids in high dose group. All results proved that the cefquinome sulfate liposomes was safe and reliable based on the animal experiments.

In order to study the differences of productive performances and serum biochemical indices between primiparous and mulitiparous dairy cows during the perinatal period, ten primiparous and ten multiparous healthy dairy cows were selected and were fed the same total mixed ration.Blood samples were collected from the caudal vein before the morning feeding on -7, -4, -2, -1, 0, 1, 2, 4, 7 and 14 d after parturition, and serum glucose, triglyceride and calcium concentrations were determined. Milk yields were recorded every day from 11 to 40 days after parturition. Milk samples were collected from each experimental cow on 30th day after parturition and milk compositions and somatic cell counts were determined. The results indicated that no significant difference was observed in serum triglyceride, milk fat, milk protein, milk lactose and milk dry matter between primiparous and multiparous dairy cows (P>0.05). Mutiparous cows had greater daily milk yield and higher milk somatic cell count than primiparous cows (P<0.05). Primiparous cows had significant higher serum glucose level on -4, 2 and 4 d after parturition (P<0.05), but significant lower serum glucose level on calving date (P<0.05) when compared with multiparous cows. Higher serum calcium concentrations were found on -2, -1 and 2 d after parturition in primiparous cows than multiparous cows (P<0.05).

With the development of intensive broiler raising, the influence of environmental factors on broiler health are increasingly prominent. Relative humidity is one of the most important indicators of broiler house environment.But in the production process of broiler farming,it is often neglected on the management of humidity in house. Neither low humidity nor high humidity is conducive to the healthy growth of broilers. It is of great significance to strengthen the monitoring of the humidity in broiler houses and study the effects of relative humidity on the health of broiler for guiding the rational control of the humidity in broiler houses and the development of healthy breeding. In this paper, we summarized and analyzed the effects of relative humidity on thermal regulating,growth performance, reproductive performance, blood parameters, meat quality and respiratory tract of broilers, and the monitor and control measures of humidity in the poultry house, aimed to provide a reference for further research on the mechanism of relative humidity affects broiler health and rational regulation of humidity in broiler house.